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Sample records for stained histological sections

  1. Acetylcholinesterase and Nissl staining in the same histological section.

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    Shipley, M T; Ennis, M; Behbehani, M M

    1989-12-18

    Acetylcholinesterase (AChE) enzyme histochemistry and Nissl staining are commonly utilized in neural architectonic studies. However, the opaque reaction deposit produced by the most commonly used AChE histochemical methods is not compatible with satisfactory Nissl staining. As a result, precise correlation of AChE and Nissl staining necessitates time-consuming comparisons of adjacent sections which may have differential shrinkage. Here, we have modified the Koelle-Friedenwald histochemical reaction for AChE by omitting the final intensification steps. The modified reaction yields a non-opaque reaction product that is selectively visualized by darkfield illumination. This non-intensified darkfield AChE (NIDA) reaction allows clear visualization of Nissl staining in the same histological section. This combined AChE-Nissl method greatly facilitates detailed correlation of enzyme and cytoarchitectonic organization.

  2. The Luna stain, an improved selective stain for detection of microsporidian spores in histologic sections

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    Peterson, Tracy S.; Spitsbergen, Jan M.; Feist, Stephen W.; Kent, Michael L.

    2014-01-01

    Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation or small spores in nuclei (i.e., Nucleospora salmonis). Hence, a variety of selective stains that enhance visualization of spores are recommended. We discovered that the Luna stain, used to highlight eosinophils, red blood cells and chitin in arthropods and other invertebrates, also stains spores of Pseudoloma neurophilia. We compared this stain to the Gram, Fite’s acid fast, Giemsa, and H&E stains on eight aquatic microsporidian organisms that were readily available in our two laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum and an unidentified microsporidian from E. sinensis, UK. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the two microsporidia from invertebrates, Steinhausia mytilovum and the unidentified microsporidian from E. sinensis. PMID:21848126

  3. Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections

    Science.gov (United States)

    Tuer, Adam; Tokarz, Danielle; Prent, Nicole; Cisek, Richard; Alami, Jennifer; Dumont, Daniel J.; Bakueva, Ludmila; Rowlands, John; Barzda, Virginijus

    2010-03-01

    Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.

  4. Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

    Science.gov (United States)

    Chen, Jianxin; Xu, Jian; Kang, Deyong; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Jiang, Xingshan

    2013-10-01

    Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

  5. Three-dimensional reconstruction of port wine stain vascular anatomy from serial histological sections

    NARCIS (Netherlands)

    Smithies, D. J.; van Gemert, M. J.; Hansen, M. K.; Milner, T. E.; Nelson, J. S.

    1997-01-01

    Port wine stains (PWSs) treated with a flashlamp-pumped pulsed dye laser show a variability in clinical response that is incompletely understood. To identify any vascular structure that might adversely affect treatment response, we obtained a three-dimensional reconstruction of the vascular anatomy

  6. Histological Stains: A Literature Review and Case Study.

    Science.gov (United States)

    Alturkistani, Hani A; Tashkandi, Faris M; Mohammedsaleh, Zuhair M

    2015-06-25

    The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures. Many stain procedures are still in use today, and many others have been replaced with new immunostaining, molecular, non-culture and other advanced staining techniques. Some staining methods have been abandoned because the chemicals required have been medically proven to be toxic. The case studies indicated that in modern histology a combination of different stain techniques are used to enhance the effectiveness of the staining process. Currently, improved histological stains, have been modified and combined with other stains to improve their effectiveness.

  7. Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA

    OpenAIRE

    Wang, Hongyang; Owens, James D; Shih, Joanna H; Li, Ming-Chung; Bonner, Robert F; Mushinski, J Frederic

    2006-01-01

    Abstract Background Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM) faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM...

  8. Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA

    Directory of Open Access Journals (Sweden)

    Li Ming-Chung

    2006-04-01

    Full Text Available Abstract Background Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E, Nissl Stain (NS, and for immunofluorescence (IF as well as with the plasma cell-revealing methyl green pyronin (MGP stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. Results The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. Conclusion RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

  9. Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA.

    Science.gov (United States)

    Wang, Hongyang; Owens, James D; Shih, Joanna H; Li, Ming-Chung; Bonner, Robert F; Mushinski, J Frederic

    2006-04-27

    Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM) faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E), Nissl Stain (NS), and for immunofluorescence (IF) as well as with the plasma cell-revealing methyl green pyronin (MGP) stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

  10. Histological identification of H. pylori stained by hematoxylin-eosin and Giemsa: review for quality control

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    Marcela S. Boldt

    2015-04-01

    Full Text Available Introduction: Several special staining methods are available for H. pylori (Hp identification in histological sections of chronic gastritis (CG, including the routine hematoxylin-eosin (HE method. Some reports suggest that ancillary stains are not always needed to establish the diagnosis of Hp infection. In addition, the benefit of using them, when biopsies show minimal inflammation, is not clear. Objective: We performed a retrospective study to compare the usefulness of HE with Giemsa method for the histopathological diagnosis of Hp in tissue sections. Methods: Histological sections from 390 consecutive patients were reviewed. The patients were registered in the histopathology laboratory of Instituto Alfa de Gastroenterologia, Hospital das Clínicas da Universidade Federal de Minas Gerais (UFMG, Belo Horizonte, Brasil. They were divided in 4 groups according to the gastric inflammatory changes as follows: Group I, gastric mucosa with normal morphology or minimal inflammatory changes (n = 146; Group II, chronic gastritis (CG with mild inflammatory activity (n = 101; Group III, CG with patent inflammatory activity (n = 123; Group IV, patients with atrophic body gastritis (n = 20. All histological sections were carefully evaluated by 2 examiners at the oil immersion objective (1000×. Results: The identification of Hp was positive by Giemsa and HE, respectively at: Group III, 111 (90.2% and 93 (75.6% patients (p < 0.01; Group II, 43 (42.6% and 29 (28.7% patients (p < 0.05. Hp was negative in Groups I and IV. Conclusion: The results show that Giemsa stain is superior to HE for histological identification of Hp in CG. Although Hp could be identified by HE stain in the majority of CG cases, a significant number of infected patients may be neglected, regardless the intensity of the inflammatory response.

  11. Blockface histology with optical coherence tomography: a comparison with Nissl staining.

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    Magnain, Caroline; Augustinack, Jean C; Reuter, Martin; Wachinger, Christian; Frosch, Matthew P; Ragan, Timothy; Akkin, Taner; Wedeen, Van J; Boas, David A; Fischl, Bruce

    2014-01-01

    Spectral domain optical coherence tomography (SD-OCT) is a high resolution imaging technique that generates excellent contrast based on intrinsic optical properties of the tissue, such as neurons and fibers. The SD-OCT data acquisition is performed directly on the tissue block, diminishing the need for cutting, mounting and staining. We utilized SD-OCT to visualize the laminar structure of the isocortex and compared cortical cytoarchitecture with the gold standard Nissl staining, both qualitatively and quantitatively. In histological processing, distortions routinely affect registration to the blockface image and prevent accurate 3D reconstruction of regions of tissue. We compared blockface registration to SD-OCT and Nissl, respectively, and found that SD-OCT-blockface registration was significantly more accurate than Nissl-blockface registration. Two independent observers manually labeled cortical laminae (e.g. III, IV and V) in SD-OCT images and Nissl stained sections. Our results show that OCT images exhibit sufficient contrast in the cortex to reliably differentiate the cortical layers. Furthermore, the modalities were compared with regard to cortical laminar organization and showed good agreement. Taken together, these SD-OCT results suggest that SD-OCT contains information comparable to standard histological stains such as Nissl in terms of distinguishing cortical layers and architectonic areas. Given these data, we propose that SD-OCT can be used to reliably generate 3D reconstructions of multiple cubic centimeters of cortex that can be used to accurately and semi-automatically perform standard histological analyses. © 2013.

  12. Short Nissl staining for incubated cryostat sections of the brain.

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    Lindroos, O F

    1991-01-01

    Nissl stain often binds poorly to cryostat sections which have been incubated in solutions of radiolabeled ligands. Such incubation is used in receptor autoradiography of the brain when using the in vitro method. We have developed a rapid (16 min) modification of Nissl staining for sections that bind stain poorly, e.g., incubated sections. The method stains well sections which cannot be stained with other rapid Nissl staining methods.

  13. A silver-stain modification of standard histological slide preparation for use in anthropology analyses.

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    Pinto, Deborrah C; Pace, Eric D

    2015-03-01

    The traditional histology method typically employed by forensic anthropologists involves plastic embedding of undecalcified bone. The embedded sample is then cut by a diamond blade saw and ground to the required thickness of ~50-100 microns using a grinder. There are several limitations to this method: Cement lines may be blurred; depth-of-field artifacts may result from viewing thick sections; and medicolegal offices with limited budgets may not be able to invest in additional equipment or training for this method. A silver nitrate stain modification of the standard histology preparation technique of decalcified bone is presented. The benefits of this technique are that: Cement lines are viewed clearly; no depth-of-field artifacts are present; and because this is a modification of the standard technique used by histology laboratories typically employed by medicolegal offices, no additional equipment or training is required. © 2015 American Academy of Forensic Sciences.

  14. Zooming in: high resolution 3D reconstruction of differently stained histological whole slide images

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    Lotz, Johannes; Berger, Judith; Müller, Benedikt; Breuhahn, Kai; Grabe, Niels; Heldmann, Stefan; Homeyer, André; Lahrmann, Bernd; Laue, Hendrik; Olesch, Janine; Schwier, Michael; Sedlaczek, Oliver; Warth, Arne

    2014-03-01

    Much insight into metabolic interactions, tissue growth, and tissue organization can be gained by analyzing differently stained histological serial sections. One opportunity unavailable to classic histology is three-dimensional (3D) examination and computer aided analysis of tissue samples. In this case, registration is needed to reestablish spatial correspondence between adjacent slides that is lost during the sectioning process. Furthermore, the sectioning introduces various distortions like cuts, folding, tearing, and local deformations to the tissue, which need to be corrected in order to exploit the additional information arising from the analysis of neighboring slide images. In this paper we present a novel image registration based method for reconstructing a 3D tissue block implementing a zooming strategy around a user-defined point of interest. We efficiently align consecutive slides at increasingly fine resolution up to cell level. We use a two-step approach, where after a macroscopic, coarse alignment of the slides as preprocessing, a nonlinear, elastic registration is performed to correct local, non-uniform deformations. Being driven by the optimization of the normalized gradient field (NGF) distance measure, our method is suitable for differently stained and thus multi-modal slides. We applied our method to ultra thin serial sections (2 μm) of a human lung tumor. In total 170 slides, stained alternately with four different stains, have been registered. Thorough visual inspection of virtual cuts through the reconstructed block perpendicular to the cutting plane shows accurate alignment of vessels and other tissue structures. This observation is confirmed by a quantitative analysis. Using nonlinear image registration, our method is able to correct locally varying deformations in tissue structures and exceeds the limitations of globally linear transformations.

  15. μCT of ex-vivo stained mouse hearts and embryos enables a precise match between 3D virtual histology, classical histology and immunochemistry

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    Larsson, Emanuel; Martin, Sabine; Lazzarini, Marcio; Tromba, Giuliana; Missbach-Guentner, Jeannine; Pinkert-Leetsch, Diana; Katschinski, Dörthe M.; Alves, Frauke

    2017-01-01

    The small size of the adult and developing mouse heart poses a great challenge for imaging in preclinical research. The aim of the study was to establish a phosphotungstic acid (PTA) ex-vivo staining approach that efficiently enhances the x-ray attenuation of soft-tissue to allow high resolution 3D visualization of mouse hearts by synchrotron radiation based μCT (SRμCT) and classical μCT. We demonstrate that SRμCT of PTA stained mouse hearts ex-vivo allows imaging of the cardiac atrium, ventricles, myocardium especially its fibre structure and vessel walls in great detail and furthermore enables the depiction of growth and anatomical changes during distinct developmental stages of hearts in mouse embryos. Our x-ray based virtual histology approach is not limited to SRμCT as it does not require monochromatic and/or coherent x-ray sources and even more importantly can be combined with conventional histological procedures. Furthermore, it permits volumetric measurements as we show for the assessment of the plaque volumes in the aortic valve region of mice from an ApoE-/- mouse model. Subsequent, Masson-Goldner trichrome staining of paraffin sections of PTA stained samples revealed intact collagen and muscle fibres and positive staining of CD31 on endothelial cells by immunohistochemistry illustrates that our approach does not prevent immunochemistry analysis. The feasibility to scan hearts already embedded in paraffin ensured a 100% correlation between virtual cut sections of the CT data sets and histological heart sections of the same sample and may allow in future guiding the cutting process to specific regions of interest. In summary, since our CT based virtual histology approach is a powerful tool for the 3D depiction of morphological alterations in hearts and embryos in high resolution and can be combined with classical histological analysis it may be used in preclinical research to unravel structural alterations of various heart diseases. PMID:28178293

  16. Taste in Art-Exposure to Histological Stains Shapes Abstract Art Preferences.

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    Böthig, Antonia M; Hayn-Leichsenring, Gregor U

    2017-01-01

    Exposure to art increases the appreciation of artworks. Here, we showed that this effect is domain independent. After viewing images of histological stains in a lecture, ratings increased for restricted subsets of abstract art images. In contrast, a lecture on art history generally enhanced ratings for all art images presented, while a lecture on town history without any visual stimuli did not increase the ratings. Therefore, we found a domain-independent exposure effect of images of histological stains to particular abstract paintings. This finding suggests that the 'taste' for abstract art is altered by visual impressions that are presented outside of an artistic context.

  17. Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin.

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    Sullivan-Brown, Jessica; Bisher, Margaret E; Burdine, Rebecca D

    2011-01-01

    Histological techniques are critical for observing tissue and cellular morphology. In this paper, we outline our protocol for embedding, serial sectioning, staining and visualizing zebrafish embryos embedded in JB-4 plastic resin-a glycol methacrylate-based medium that results in excellent preservation of tissue morphology. In addition, we describe our procedures for staining plastic sections with toluidine blue or hematoxylin and eosin, and show how to couple these stains with whole-mount RNA in situ hybridization. We also describe how to maintain and visualize immunofluorescence and EGFP signals in JB-4 resin. The protocol we outline-from embryo preparation, embedding, sectioning and staining to visualization-can be accomplished in 3 d. Overall, we reinforce that plastic embedding can provide higher resolution of cellular details and is a valuable tool for cellular and morphological studies in zebrafish.

  18. Standardization of reagents and methods used in cytological and histological practice with emphasis on dyes, stains and chromogenic reagents

    DEFF Research Database (Denmark)

    Lyon, H O; De Leenheer, A P; Horobin, R W

    1994-01-01

    and stains through the preparation of reference materials and the development of chromatographic methods. An overview is presented of the problems concerned with standardization of the Romanowsky-Giemsa stain for cytological and histological application. Finally, the problem of how to convince routine dye...

  19. Demonstration of lipofuscin and Nissl bodies in crystal violet stained sections using a fluorescence technique or pyronin Y stain.

    Science.gov (United States)

    Terr, L I

    1986-09-01

    This paper presents two simple, reliable methods for identification of lipofuscin and Nissl bodies in the same section. One method shows that lipofuscin stained with crystal violet retains its ability to fluoresce and can be observed under the fluorescence microscope after the stain has faded. Fading is accompanied by a gradual increase in the intensity of the fluorescence and is complete in about 5 min. Exciting illumination from this part of the spectrum also substantially fades staining of other autofluorescing tissue elements, such as lipids. Nonfluorescing structures, such as Nissl bodies, remain stained. By changing from transillumination with tungsten light to epifluorescent illumination and vice versa, both types of structures--Nissl bodies and lipofuscin--can be identified in the same section. The second technique uses pyronin Y for staining Nissl bodies in preparations previously stained with crystal violet. Nissl bodies are stained pink but lipofuscin remains violet. Lipofuscin in these sections also remains autofluorescent after the crystal violet stain has faded under violet or near-UV light.

  20. 3D prostate histology image reconstruction: Quantifying the impact of tissue deformation and histology section location

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    Eli Gibson

    2013-01-01

    Full Text Available Background: Guidelines for localizing prostate cancer on imaging are ideally informed by registered post-prostatectomy histology. 3D histology reconstruction methods can support this by reintroducing 3D spatial information lost during histology processing. The need to register small, high-grade foci drives a need for high accuracy. Accurate 3D reconstruction method design is impacted by the answers to the following central questions of this work. (1 How does prostate tissue deform during histology processing? (2 What spatial misalignment of the tissue sections is induced by microtome cutting? (3 How does the choice of reconstruction model affect histology reconstruction accuracy? Materials and Methods: Histology, paraffin block face and magnetic resonance images were acquired for 18 whole mid-gland tissue slices from six prostates. 7-15 homologous landmarks were identified on each image. Tissue deformation due to histology processing was characterized using the target registration error (TRE after landmark-based registration under four deformation models (rigid, similarity, affine and thin-plate-spline [TPS]. The misalignment of histology sections from the front faces of tissue slices was quantified using manually identified landmarks. The impact of reconstruction models on the TRE after landmark-based reconstruction was measured under eight reconstruction models comprising one of four deformation models with and without constraining histology images to the tissue slice front faces. Results: Isotropic scaling improved the mean TRE by 0.8-1.0 mm (all results reported as 95% confidence intervals, while skew or TPS deformation improved the mean TRE by <0.1 mm. The mean misalignment was 1.1-1.9΀ (angle and 0.9-1.3 mm (depth. Using isotropic scaling, the front face constraint raised the mean TRE by 0.6-0.8 mm. Conclusions: For sub-millimeter accuracy, 3D reconstruction models should not constrain histology images to the tissue slice front faces and

  1. Combined in situ zymography, immunofluorescence, and staining of iron oxide particles in paraffin-embedded, zinc-fixed tissue sections.

    Science.gov (United States)

    Haeckel, Akvile; Schoenzart, Lena; Appler, Franziska; Schnorr, Joerg; Taupitz, Matthias; Hamm, Bernd; Schellenberger, Eyk

    2012-01-01

    Superparamagnetic iron oxide particles are used as potent contrast agents in magnetic resonance imaging. In histology, these particles are frequently visualized by Prussian blue iron staining of aldehyde-fixed, paraffin-embedded tissues. Recently, zinc salt-based fixative was shown to preserve enzyme activity in paraffin-embedded tissues. In this study, we demonstrate that zinc fixation allows combining in situ zymography with fluorescence immunohistochemistry (IHC) and iron staining for advanced biologic investigation of iron oxide particle accumulation. Very small iron oxide particles, developed for magnetic resonance angiography, were applied intravenously to BALB/c nude mice. After 3 hours, spleens were explanted and subjected to zinc fixation and paraffin embedding. Cut tissue sections were further processed to in situ zymography, IHC, and Prussian blue staining procedures. The combination of in situ zymography as well as IHC with subsequent Prussian blue iron staining on zinc-fixed paraffin-embedded tissues resulted in excellent histologic images of enzyme activity, protease distribution, and iron oxide particle accumulation. The combination of all three stains on a single section allowed direct comparison with only moderate degradation of fluorescein isothiocyanate-labeled substrate. This protocol is useful for investigating the biologic environment of accumulating iron oxide particles, with excellent preservation of morphology.

  2. Fungal Fluorescence in Hematoxylin-Eosin Stained Sections

    Directory of Open Access Journals (Sweden)

    Murat Durdu

    2017-06-01

    Full Text Available A forty-six-year-old male presented to our dermatology clinic with two-year history of itching on his groin. His medical history revealed various topical corticosteroid creams without improvement of the skin lesion. Dermatological examination revealed erythematous nodules and follicular pustules on erythematous background on the inguinal area (Figure 1a. Potassium hydroxide (KOH examination was negative. Tzanck smear revealed abundant neutrophils without bacteria, fungi, or parasite. The histopathological examination showed granuloma formation with multinuclear giant cells and Periodic acid-Schiff (PAS-positive hyphae and spores around the hair follicles (Figure 1b, 1c. Hematoxylin-eosin (H&E-stained slides were examined under an immunofluorescence microscope, and these hyphae and spores showed autofluorescence (Figure 1d. Based on the clinical and histopathological findings, a Majocchi’s granuloma was considered. All lesions disappeared with topical and systemic terbinafine (250 mg/day treatment for six weeks.\tPearls;\tClinical: Not only bacteria, but also fungi, parasites, and viruses may cause folliculitis. Cytology should be initially done to identify the causes of infectious folliculitis. In case of negative cytology, histopathological examination and molecular methods can be used.\tCytological: To cytologically identify all of the causes of folliculitis, four separate samples should be taken: (i the first sample is stained with May-Grünwald-Giemsa for routine cytological examination; (ii the second sample is used for KOH testing; (iii the third sample is stained with an acid-fast stain to detect mycobacteria; and (iv the last specimen is Gram-stained to identify whether it is Gram-positive or Gram-negative (1. Histopathological: In infectious diseases, a definitive diagnosis should be done to identify the etiologic agent. The detection of fungal elements is challenging, when histopathological examination is performed with the H

  3. Electrochemical removal of metallic implants from Technovit 9100 New embedded hard and soft tissues prior to histological sectioning.

    Science.gov (United States)

    Willbold, Elmar; Reebmann, Mattias; Jeffries, Richard; Witte, Frank

    2013-11-01

    Solid metallic implants in soft or hard tissues are serious challenges for histological processing. However, metallic implants are more frequently used in e.g. cardiovascular or orthopaedic therapies. Before clinical use, these devices need to be tested thoroughly in a biological environment and histological analysis of their biocompatibility is a major requirement. To allow the histological analysis of metallic implants in tissues especially in calcified hard tissues, we describe a method for embedding these tissues in the resin Technovit 9100 New and removing the metallic implants by electrochemical dissolution. With the combination of these two processes, we are able to achieve 5 μm thick sections from soft or hard tissues with a superior preservation of tissue architecture and especially the implant-tissue interface. These sections can be stained by classical stainings, immunohistochemical and enzymehistochemical as well as DNA-based staining methods.

  4. AN EVALUATION OF THREE CONVENTIONAL HISTOLOGICAL TECHNIQUES FOR STAINING THE CERATA OF CRATENA PILATA

    Science.gov (United States)

    Fixation and staining methods for different types of tissue in the marine nudibranch Cratena pilata were evaluated. Cratena pilata, a marine snail in the Phylum Mollusca, has the ability to take stinging cells, called nematocysts, from ingested animals belonging to another phylum...

  5. Photoacoustic microscopy enables multilayered histological imaging of human breast cancer without staining

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    Wong, Terence T. W.; Zhang, Ruiying; Hai, Pengfei; Aft, Rebecca L.; Novack, Deborah V.; Wang, Lihong V.

    2018-02-01

    In 2016, an estimated 250,000 new cases of invasive and non-invasive breast cancer were diagnosed in US women. About 60-75% of these cases were treated with breast conserving surgery (BCS) as the initial therapy. To reduce the local recurrence rate, the goal of BCS is to excise the tumor with a rim of normal surrounding tissue, so that no cancer cells remain at the cut margin, while preserving as much normal breast tissue as possible. Therefore, patients with remaining cancer cells at the cut margin commonly require a second surgical procedure to obtain clear margins. Different approaches have been used to decrease the positive margin rate to avoid re-excision. However, these techniques are variously ineffective in reducing the re-operative rate, difficult to master by surgeons, or time-consuming for large specimens. Thus, 20-60% of patients undergoing BCS still require second surgeries due to positive surgical margins. The ideal tool for margin assessment would provide the same information as histological analysis, without the need for processing specimens. To achieve this goal, we have developed and refined label-free photoacoustic microscopy (PAM) for breast specimens. Exploiting the intrinsic optical contrast of tissue, ultraviolet (UV) laser illumination can highlight cell nuclei, thus providing the same contrast as hematoxylin labeling used in conventional histology and measuring features related to the histological landscape without the need for labels. We demonstrate that our UV-PAM system can provide label-free, high-resolution, and histology-like imaging of fixed, unprocessed breast tissue.

  6. Design-based stereological estimation of the total number of cardiac myocytes in histological sections

    DEFF Research Database (Denmark)

    Brüel, Annemarie; Nyengaard, Jens Randel

    2005-01-01

    in LM sections using design-based stereology. MATERIALS AND METHODS: From formalin-fixed left rat ventricles (LV) isotropic uniformly random sections were cut. The total number of myocyte nuclei per LV was estimated using the optical disector. Two-microm-thick serial paraffin sections were stained......BACKGROUND: Counting the total number of cardiac myocytes has not previously been possible in ordinary histological sections using light microscopy (LM) due to difficulties in defining the myocyte borders properly. AIM: To describe a method by which the total number of cardiac myocytes is estimated...... with antibodies against cadherin and type IV collagen to visualise the intercalated discs and the myocyte membranes, respectively. Using the physical disector in "local vertical windows" of the serial sections, the average number of nuclei per myocyte was estimated.RESULTS: The total number of myocyte nuclei...

  7. Preparation of High-quality Hematoxylin and Eosin-stained Sections from Rodent Mammary Gland Whole Mounts for Histopathologic Review.

    Science.gov (United States)

    Tucker, Deirdre K; Foley, Julie F; Hayes-Bouknight, Schantel A; Fenton, Suzanne E

    2016-10-01

    Identifying environmental exposures that cause adverse mammary gland outcomes in rodents is a first step in disease prevention in humans and domestic pets. "Whole mounts" are an easy and inexpensive tissue preparation method that can elucidate typical or abnormal mammary gland morphology in rodent studies. Here, we propose procedures to facilitate the use of whole mounts for histological identification of grossly noted tissue alterations. We noted lesions in mammary whole mounts from 14-month-old CD-1 mice that were not found in the contralateral gland hematoxylin and eosin (H&E)-stained section. Whole mounts were removed from the slide and carefully processed to produce high-quality histological sections that mirrored the quality of the original H&E-stained section in order to properly diagnose the unidentified gross abnormalities. Incorporation of this method into testing protocols that focus on human relevant chemical and endocrine disruptors exposure will increase the chances of identifying lesions in the gland and reduce the risk of false negative findings. This method can be especially invaluable when lesions are not always palpable during the course of the study or visible at necropsy, or when a single cross section of the mammary gland is otherwise used for detecting lesions. © The Author(s) 2016.

  8. When machine vision meets histology: A comparative evaluation of model architecture for classification of histology sections.

    Science.gov (United States)

    Zhong, Cheng; Han, Ju; Borowsky, Alexander; Parvin, Bahram; Wang, Yunfu; Chang, Hang

    2017-01-01

    Classification of histology sections in large cohorts, in terms of distinct regions of microanatomy (e.g., stromal) and histopathology (e.g., tumor, necrosis), enables the quantification of tumor composition, and the construction of predictive models of genomics and clinical outcome. To tackle the large technical variations and biological heterogeneities, which are intrinsic in large cohorts, emerging systems utilize either prior knowledge from pathologists or unsupervised feature learning for invariant representation of the underlying properties in the data. However, to a large degree, the architecture for tissue histology classification remains unexplored and requires urgent systematical investigation. This paper is the first attempt to provide insights into three fundamental questions in tissue histology classification: I. Is unsupervised feature learning preferable to human engineered features? II. Does cellular saliency help? III. Does the sparse feature encoder contribute to recognition? We show that (a) in I, both Cellular Morphometric Feature and features from unsupervised feature learning lead to superior performance when compared to SIFT and [Color, Texture]; (b) in II, cellular saliency incorporation impairs the performance for systems built upon pixel-/patch-level features; and (c) in III, the effect of the sparse feature encoder is correlated with the robustness of features, and the performance can be consistently improved by the multi-stage extension of systems built upon both Cellular Morphmetric Feature and features from unsupervised feature learning. These insights are validated with two cohorts of Glioblastoma Multiforme (GBM) and Kidney Clear Cell Carcinoma (KIRC). Copyright © 2016 Elsevier B.V. All rights reserved.

  9. [Immunocytochemical demonstration of astrocytes in brain sections combined with Nissl staining].

    Science.gov (United States)

    Korzhevskiĭ, D E; Otellin, V A

    2004-01-01

    The aim of the present study was to develop an easy and reliable protocol of combined preparation staining, which would unite the advantages of immunocytochemical demonstration of astrocytes with the availability to evaluate functional state of neurons provided by Nissl technique. The presented protocol of paraffin sections processing allows to retain high quality of tissue structure and provides for selective demonstration of astrocytes using the monoclonal antibodies against glial fibrillary acidic protein and contrast Nissl staining of cells. The protocol can be used without any changes for processing of brain sections obtained from the humans and other mammals with the exception of mice and rabbits.

  10. Analysis of road traffic crash injuries - a technique producing large un-decalcified histological sections

    DEFF Research Database (Denmark)

    Uhrenholt, Lars; Gregersen, Markil Ebbe Gregers; Vesterby, Annie

    Introduction:The lower cervical spine facet joints are important structures in cases of chronic pain syndromes following road traffic crashes. Pathophysiological segmental kinematics may occur, particularly during rear-impact collisions, which may cause injury to these joints. Detailed anatomical...... that prepares large un-frozen un-decalcified cervical spine specimens for analysis. Materials and Methods:The cervical spine segments from C4 to C7 are removed en bloc during autopsy. The specimen is fixed throughout in 70% increasing to 99% ethanol and embedded un-decalcified in hardening methyl methacrylate......-duty microtome followed by relevant staining.Results:The described method produces fine detail histological sections that may visualise normal anatomical structures as well as discrete pathoanatomical lesions in the lower cervical spine facet joints.Discussion:The cervical spine facet joints can be analysed...

  11. A method for acetylcholinesterase staining of brain sections previously processed for receptor autoradiography.

    Science.gov (United States)

    Lim, M M; Hammock, E A D; Young, L J

    2004-02-01

    Receptor autoradiography using selective radiolabeled ligands allows visualization of brain receptor distribution and density on film. The resolution of specific brain regions on the film often can be difficult to discern owing to the general spread of the radioactive label and the lack of neuroanatomical landmarks on film. Receptor binding is a chemically harsh protocol that can render the tissue virtually unstainable by Nissl and other conventional stains used to delineate neuroanatomical boundaries of brain regions. We describe a method for acetylcholinesterase (AChE) staining of slides previously processed for receptor binding. AChE staining is a useful tool for delineating major brain nuclei and tracts. AChE staining on sections that have been processed for receptor autoradiography provides a direct comparison of brain regions for more precise neuroanatomical description. We report a detailed thiocholine protocol that is a modification of the Koelle-Friedenwald method to amplify the AChE signal in brain sections previously processed for autoradiography. We also describe several temporal and experimental factors that can affect the density and clarity of the AChE signal when using this protocol.

  12. Examination of Cervical Spine Histological Sections - A Technical Note

    DEFF Research Database (Denmark)

    Uhrenholt, Lars; Ullerup, Rita; Vesterby, Annie

    2006-01-01

    Detailed knowledge of the cervical spine facet joints morphology and anatomy is increasingly important since improved understanding of clinical syndromes, such as whiplash injuries, and therapeutic interventions is based on this knowledge. So far systematic examination of the age-related morphology...... of these joints has not yet been performed, nor has any generally accepted histological classification system for degenerative changes in the cervical spine facet joints been proposed. In the case of whiplash injuries the presented histological method has particular relevance since it allows detailed description...

  13. Staining of E-selectin ligands on paraffin-embedded sections of tumor tissue.

    Science.gov (United States)

    Carrascal, Mylène A; Talina, Catarina; Borralho, Paula; Gonçalo Mineiro, A; Henriques, Ana Raquel; Pen, Cláudia; Martins, Manuela; Braga, Sofia; Sackstein, Robert; Videira, Paula A

    2018-05-02

    The E-selectin ligands expressed by cancer cells mediate adhesion of circulating cancer cells to endothelial cells, as well as within tissue microenvironments important for tumor progression and metastasis. The identification of E-selectin ligands within cancer tissue could yield new biomarkers for patient stratification and aid in identifying novel therapeutic targets. The determinants of selectin ligands consist of sialylated tetrasaccharides, the sialyl Lewis X and A (sLe X and sLe A ), displayed on protein or lipid scaffolds. Standardized procedures for immunohistochemistry make use of the antibodies against sLe X and/or sLe A . However, antibody binding does not define E-selectin binding activity. In this study, we developed an immunohistochemical staining technique, using E-selectin-human Ig Fc chimera (E-Ig) to characterize the expression and localization of E-selectin binding sites on paraffin-embedded sections of different cancer tissue. E-Ig successfully stained cancer cells with high specificity. The E-Ig staining show high reactivity scores in colon and lung adenocarcinoma and moderate reactivity in triple negative breast cancer. Compared with reactivity of antibody against sLe X/A , the E-Ig staining presented higher specificity to cancer tissue with better defined borders and less background. The E-Ig staining technique allows the qualitative and semi-quantitative analysis of E-selectin binding activity on cancer cells. The development of accurate techniques for detection of selectin ligands may contribute to better diagnostic and better understanding of the molecular basis of tumor progression and metastasis.

  14. Examination of Cervical Spine Histological Sections - A Technical Note

    DEFF Research Database (Denmark)

    Uhrenholt, Lars; Ullerup, Rita; Vesterby, Annie

    2006-01-01

    Detailed knowledge of the cervical spine facet joints morphology and anatomy is increasingly important since improved understanding of clinical syndromes, such as whiplash injuries, and therapeutic interventions is based on this knowledge. So far systematic examination of the age-related morphology...... of these joints has not yet been performed, nor has any generally accepted histological classification system for degenerative changes in the cervical spine facet joints been proposed. In the case of whiplash injuries the presented histological method has particular relevance since it allows detailed description...... of the anatomy and pathoanatomical status of the osteo-cartilagenous structures, including the facet joints from where a major portion of chronic whiplash patients experience their pain symptoms....

  15. Confocal Microscopy of Unfixed Breast Needle Core Biopsies: A Comparison to Fixed and Stained Sections

    Directory of Open Access Journals (Sweden)

    Zavislan James M

    2009-08-01

    Full Text Available Abstract Background Needle core biopsy, often in conjunction with ultrasonic or stereotactic guided techniques, is frequently used to diagnose breast carcinoma in women. Confocal scanning laser microscopy (CSLM is a technology that provides real-time digital images of tissues with cellular resolution. This paper reports the progress in developing techniques to rapidly screen needle core breast biopsy and surgical specimens at the point of care. CSLM requires minimal tissue processing and has the potential to reduce the time from excision to diagnosis. Following imaging, specimens can still be submitted for standard histopathological preparation. Methods Needle core breast specimens from 49 patients were imaged at the time of biopsy. These lesions had been characterized under the Breast Imaging Reporting And Data System (BI-RADS as category 3, 4 or 5. The core biopsies were imaged with the CSLM before fixation. Samples were treated with 5% citric acid and glycerin USP to enhance nuclear visibility in the reflectance confocal images. Immediately following imaging, the specimens were fixed in buffered formalin and submitted for histological processing and pathological diagnosis. CSLM images were then compared to the standard histology. Results The pathologic diagnoses by standard histology were 7 invasive ductal carcinomas, 2 invasive lobular carcinomas, 3 ductal carcinomas in-situ (CIS, 21 fibrocystic changes/proliferative conditions, 9 fibroadenomas, and 5 other/benign; two were excluded due to imaging difficulties. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images and were comparable to standard histologic sections of the same tissue. Conclusion CSLM is a technique with the potential to screen needle core biopsy specimens in real-time. The confocal images contained sufficient information to identify stromal reactions such as fibrosis and cellular proliferations such as intra-ductal and

  16. Confocal Microscopy of Unfixed Breast Needle Core Biopsies: A Comparison to Fixed and Stained Sections

    Science.gov (United States)

    2009-01-01

    Background Needle core biopsy, often in conjunction with ultrasonic or stereotactic guided techniques, is frequently used to diagnose breast carcinoma in women. Confocal scanning laser microscopy (CSLM) is a technology that provides real-time digital images of tissues with cellular resolution. This paper reports the progress in developing techniques to rapidly screen needle core breast biopsy and surgical specimens at the point of care. CSLM requires minimal tissue processing and has the potential to reduce the time from excision to diagnosis. Following imaging, specimens can still be submitted for standard histopathological preparation. Methods Needle core breast specimens from 49 patients were imaged at the time of biopsy. These lesions had been characterized under the Breast Imaging Reporting And Data System (BI-RADS) as category 3, 4 or 5. The core biopsies were imaged with the CSLM before fixation. Samples were treated with 5% citric acid and glycerin USP to enhance nuclear visibility in the reflectance confocal images. Immediately following imaging, the specimens were fixed in buffered formalin and submitted for histological processing and pathological diagnosis. CSLM images were then compared to the standard histology. Results The pathologic diagnoses by standard histology were 7 invasive ductal carcinomas, 2 invasive lobular carcinomas, 3 ductal carcinomas in-situ (CIS), 21 fibrocystic changes/proliferative conditions, 9 fibroadenomas, and 5 other/benign; two were excluded due to imaging difficulties. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images and were comparable to standard histologic sections of the same tissue. Conclusion CSLM is a technique with the potential to screen needle core biopsy specimens in real-time. The confocal images contained sufficient information to identify stromal reactions such as fibrosis and cellular proliferations such as intra-ductal and infiltrating carcinoma, and

  17. Diagnostic accuracy of morphologic identification of filamentous fungi in paraffin embedded tissue sections: Correlation of histological and culture diagnosis

    Directory of Open Access Journals (Sweden)

    Sundaram Challa

    2014-01-01

    Full Text Available Aims and Objectives: The aim was to investigate the correlation between histological and culture diagnosis of filamentous fungi. Materials and Methods: Tissue sections from biopsy samples stained with Hematoxylin and Eosin and special stains from samples of chronic invasive/noninvasive sinusitis and intracranial space occupying lesions during 2005-2011 diagnosed to have infection due to filamentous fungi were reviewed. The histopathology and culture diagnoses were analyzed for correlation and discrepancy. Results: There were 125 samples positive for filamentous fungi on biopsy. Of these 76 (60.8% were submitted for culture and fungi grew in 30 (39.97% samples. There was a positive correlation between histological and culture diagnosis in 25 (83.33% samples that included Aspergillus species (16/19, Zygomycetes species (8/10 and dematiaceous fungi (1/1. The negative yield of fungi was more in Zygomycetes species (20/30 when compared to Aspergillus species (25/44. There was a discrepancy in diagnosis in 5/30 (16.67% samples which included probable dual infection in two, and dematiaceous fungi being interpreted as Aspergillus species in three samples. Conclusion: Histopathology plays a major role in the diagnosis of infection due to filamentous fungi, especially when cultures are not submitted or negative. The discrepancy between histological and culture diagnosis was either due to dematiaceous fungi being interpreted as Aspergillus species or probable dual infection.

  18. Generation of a 3D atlas of the nuclear division of the thalamus based on histological sections of primate: Intra- and intersubject atlas-to-MRI warping

    International Nuclear Information System (INIS)

    Dauguet, J.; Conde, F.; Hantraye, P.; Delzescaux, T.; Frouin, V.

    2009-01-01

    We describe a framework to generate a 3D digital atlas of the thalamus based on a series of stained histological sections of a primate. The contours of the thalamus were first drawn on the stained histological slices. The series of histological sections were then aligned and mapped onto the in vivo MRI of the same animal acquired prior to the sacrifice following a methodology described in Dauguet et al. (2007) using the block face photographs as an intermediary modality. By applying the series of transformations previously estimated for the histological volume, the contours of the digital atlas were mapped onto the MRI data. The protocol was tested on two baboon brains for which the full series of slices were available, and a macaque brain for which a subset only of the histological slices were available demonstrating the ability of building digital atlases in the MRI geometry without mounting and staining all the brain slices. We then studied the accuracy of mapping the digital atlas of one baboon onto the MRI of the other baboon by comparing the overlapping with its original digital atlas. We finally used the digital atlas of one of the baboons to study the individual kinetic of the main thalamus nuclei on Positron Emission Tomography (PET) images providing a novel and accurate way of measuring very fine and local functional differences. (authors)

  19. Does amnioinfusion reduce caesarean section rate in meconium-stained amniotic fluid.

    Science.gov (United States)

    Choudhary, Deepti; Bano, Imam; Ali, S M

    2010-07-01

    The purpose of our study was to evaluate the safety and efficacy of transcervical amnioinfusion during labour complicated by meconium-stained amniotic fluid, in a setting with limited peripartum facilities, to lower the incidence of caesarean section. A prospective study was conducted in a teaching hospital in north India, which enrolled 292 patients admitted in labour. Patients were randomly divided into two groups after taking their consent. One group received transcervical amnioinfusion, whilst in the other group amnioinfusion was not done. Caesarean sections were performed in either group if there were foetal heart rate abnormalities (bradycardia or irregularity for 10-20 min) or slow progress of labour. The outcomes studied were the incidence of caesarean sections, duration of maternal hospital stay, maternal febrile morbidity (temperature of >38 degrees C, 24 h after delivery), low Apgar score (at 1 and 5 min), respiratory death, MAS and perinatal mortality. There was a statistically significant reduction in the incidence of caesarean sections in the study group compared to the control group (31 vs. 61%). Amnioinfusion was associated with improved neonatal outcome as evidenced by statistically improved Apgar score at 1 min in newborns in the study group compared to the control group (10 vs. 37.2%). Amnioinfusion during labour was not associated with any significant maternal and neonatal complications. The mean hospital stay of the mother was decreased significantly in the study group patients compared to the control group. Transcervical amnioinfusion in labour for meconium-stained amniotic fluid is a simple, safe and easy-to-perform procedure. It can be performed safely in a setup with limited peripartum facilities, especially in developing countries, to decrease intrapartum operative intervention and reduce foetomaternal morbidity and mortality.

  20. New methods for multimodal MS imaging of histological tissue sections

    NARCIS (Netherlands)

    Amstalden Van Hove, E.R.

    2011-01-01

    The insights derived from spatial localization of molecules in tissue sections are of great value for understanding and treating cancer and other diseases. These insights can relate to molecules linked to a disease as well as to drug molecules distributed across organs of interest. Mass spectrometry

  1. Evaporation process in histological tissue sections for neutron autoradiography.

    Science.gov (United States)

    Espector, Natalia M; Portu, Agustina; Santa Cruz, Gustavo A; Saint Martin, Gisela

    2018-05-01

    The analysis of the distribution and density of nuclear tracks forming an autoradiography in a nuclear track detector (NTD) allows the determination of 10 B atoms concentration and location in tissue samples from Boron Neutron Capture Therapy (BNCT) protocols. This knowledge is of great importance for BNCT dosimetry and treatment planning. Tissue sections studied with this technique are obtained by cryosectioning frozen tissue specimens. After the slicing procedure, the tissue section is put on the NTD and the sample starts drying. The thickness varies from its original value allowing more particles to reach the detector and, as the mass of the sample decreases, the boron concentration in the sample increases. So in order to determine the concentration present in the hydrated tissue, the application of corrective coefficients is required. Evaporation mechanisms as well as various factors that could affect the process of mass variation are outlined in this work. Mass evolution for tissue samples coming from BDIX rats was registered with a semimicro analytical scale and measurements were analyzed with software developed to that end. Ambient conditions were simultaneously recorded, obtaining reproducible evaporation curves. Mathematical models found in the literature were applied for the first time to this type of samples and the best fit of the experimental data was determined. The correlation coefficients and the variability of the parameters were evaluated, pointing to Page's model as the one that best represented the evaporation curves. These studies will contribute to a more precise assessment of boron concentration in tissue samples by the Neutron Autoradiography technique.

  2. Analysis of thick brain sections by obverse-reverse computer microscopy: application of a new, high clarity Golgi-Nissl stain.

    Science.gov (United States)

    Glaser, E M; Van der Loos, H

    1981-08-01

    Exceptionally clear Golgi-Nissl sections of 300 micron thickness have been morphometrically studied by light microscopy using oil immersion objectives. The clarity results from a new variation of a staining procedure that combines Golgi and Nissl images in one section. A viewing technique has been developed that permits a histologic preparation to be examined from its obverse (or normally viewed) side and its reverse (or under) side. The technique was designed for use with a computer microscope but can be employed with any light microscope whose stage position can be measured within 100 micron. Sections thicker than 300 micron can be studied dependent on the working distance of the objective lens, provided that the clarity of the material permits it.

  3. Quantifying anisotropy and fiber orientation in human brain histological sections

    Directory of Open Access Journals (Sweden)

    Matthew D Budde

    2013-02-01

    Full Text Available Diffusion weighted imaging (DWI has provided unparalleled insight into the microscopic structure and organization of the central nervous system. Diffusion tensor imaging (DTI and other models of the diffusion MRI signal extract microstructural properties of tissues with relevance to the normal and injured brain. Despite the prevalence of such techniques and applications, accurate and large-scale validation has proven difficult, particularly in the human brain. In this report, human brain sections obtained from a digital public brain bank were employed to quantify anisotropy and fiber orientation using structure tensor analysis. The derived maps depict the intricate complexity of white matter fibers at a resolution not attainable with current DWI experiments. Moreover, the effects of multiple fiber bundles (i.e. crossing fibers and intravoxel fiber dispersion were demonstrated. Examination of the cortex and hippocampal regions validated specific features of previous in vivo and ex vivo DTI studies of the human brain. Despite the limitation to two dimensions, the resulting images provide a unique depiction of white matter organization at resolutions currently unattainable with DWI. The method of analysis may be used to validate tissue properties derived from DTI and alternative models of the diffusion signal.

  4. Optical histology: a method to visualize microvasculature in thick tissue sections of mouse brain.

    Directory of Open Access Journals (Sweden)

    Austin J Moy

    Full Text Available The microvasculature is the network of blood vessels involved in delivering nutrients and gases necessary for tissue survival. Study of the microvasculature often involves immunohistological methods. While useful for visualizing microvasculature at the µm scale in specific regions of interest, immunohistology is not well suited to visualize the global microvascular architecture in an organ. Hence, use of immunohistology precludes visualization of the entire microvasculature of an organ, and thus impedes study of global changes in the microvasculature that occur in concert with changes in tissue due to various disease states. Therefore, there is a critical need for a simple, relatively rapid technique that will facilitate visualization of the microvascular network of an entire tissue.The systemic vasculature of a mouse is stained with the fluorescent lipophilic dye DiI using a method called "vessel painting". The brain, or other organ of interest, is harvested and fixed in 4% paraformaldehyde. The organ is then sliced into 1 mm sections and optically cleared, or made transparent, using FocusClear, a proprietary optical clearing agent. After optical clearing, the DiI-labeled tissue microvasculature is imaged using confocal fluorescence microscopy and adjacent image stacks tiled together to produce a depth-encoded map of the microvasculature in the tissue slice. We demonstrated that the use of optical clearing enhances both the tissue imaging depth and the estimate of the vascular density. Using our "optical histology" technique, we visualized microvasculature in the mouse brain to a depth of 850 µm.Presented here are maps of the microvasculature in 1 mm thick slices of mouse brain. Using combined optical clearing and optical imaging techniques, we devised a methodology to enhance the visualization of the microvasculature in thick tissues. We believe this technique could potentially be used to generate a three-dimensional map of the

  5. Practicable methods for histological section thickness measurement in quantitative stereological analyses.

    Science.gov (United States)

    Matenaers, Cyrill; Popper, Bastian; Rieger, Alexandra; Wanke, Rüdiger; Blutke, Andreas

    2018-01-01

    The accuracy of quantitative stereological analysis tools such as the (physical) disector method substantially depends on the precise determination of the thickness of the analyzed histological sections. One conventional method for measurement of histological section thickness is to re-embed the section of interest vertically to its original section plane. The section thickness is then measured in a subsequently prepared histological section of this orthogonally re-embedded sample. However, the orthogonal re-embedding (ORE) technique is quite work- and time-intensive and may produce inaccurate section thickness measurement values due to unintentional slightly oblique (non-orthogonal) positioning of the re-embedded sample-section. Here, an improved ORE method is presented, allowing for determination of the factual section plane angle of the re-embedded section, and correction of measured section thickness values for oblique (non-orthogonal) sectioning. For this, the analyzed section is mounted flat on a foil of known thickness (calibration foil) and both the section and the calibration foil are then vertically (re-)embedded. The section angle of the re-embedded section is then calculated from the deviation of the measured section thickness of the calibration foil and its factual thickness, using basic geometry. To find a practicable, fast, and accurate alternative to ORE, the suitability of spectral reflectance (SR) measurement for determination of plastic section thicknesses was evaluated. Using a commercially available optical reflectometer (F20, Filmetrics®, USA), the thicknesses of 0.5 μm thick semi-thin Epon (glycid ether)-sections and of 1-3 μm thick plastic sections (glycolmethacrylate/ methylmethacrylate, GMA/MMA), as regularly used in physical disector analyses, could precisely be measured within few seconds. Compared to the measured section thicknesses determined by ORE, SR measures displayed less than 1% deviation. Our results prove the applicability

  6. Identification of histological correlates of overall survival in lower grade gliomas using a bag-of-words paradigm: A preliminary analysis based on hematoxylin & eosin stained slides from the lower grade glioma cohort of the cancer genome Atlas

    Directory of Open Access Journals (Sweden)

    Reid Trenton Powell

    2017-01-01

    Full Text Available Background: Glioma, the most common primary brain neoplasm, describes a heterogeneous tumor of multiple histologic subtypes and cellular origins. At clinical presentation, gliomas are graded according to the World Health Organization guidelines (WHO, which reflect the malignant characteristics of the tumor based on histopathological and molecular features. Lower grade diffuse gliomas (LGGs (WHO Grade II–III have fewer malignant characteristics than high-grade gliomas (WHO Grade IV, and a better clinical prognosis, however, accurate discrimination of overall survival (OS remains a challenge. In this study, we aimed to identify tissue-derived image features using a machine learning approach to predict OS in a mixed histology and grade cohort of lower grade glioma patients. To achieve this aim, we used H and E stained slides from the public LGG cohort of The Cancer Genome Atlas (TCGA to create a machine learned dictionary of “image-derived visual words” associated with OS. We then evaluated the combined efficacy of using these visual words in predicting short versus long OS by training a generalized machine learning model. Finally, we mapped these predictive visual words back to molecular signaling cascades to infer potential drivers of the machine learned survival-associated phenotypes. Methods: We analyzed digitized histological sections downloaded from the LGG cohort of TCGA using a bag-of-words approach. This method identified a diverse set of histological patterns that were further correlated with OS, histology, and molecular signaling activity using Cox regression, analysis of variance, and Spearman correlation, respectively. A support vector machine (SVM model was constructed to discriminate patients into short and long OS groups dichotomized at 24-month. Results: This method identified disease-relevant phenotypes associated with OS, some of which are correlated with disease-associated molecular pathways. From these image

  7. Identification of Histological Correlates of Overall Survival in Lower Grade Gliomas Using a Bag-of-words Paradigm: A Preliminary Analysis Based on Hematoxylin & Eosin Stained Slides from the Lower Grade Glioma Cohort of The Cancer Genome Atlas.

    Science.gov (United States)

    Powell, Reid Trenton; Olar, Adriana; Narang, Shivali; Rao, Ganesh; Sulman, Erik; Fuller, Gregory N; Rao, Arvind

    2017-01-01

    Glioma, the most common primary brain neoplasm, describes a heterogeneous tumor of multiple histologic subtypes and cellular origins. At clinical presentation, gliomas are graded according to the World Health Organization guidelines (WHO), which reflect the malignant characteristics of the tumor based on histopathological and molecular features. Lower grade diffuse gliomas (LGGs) (WHO Grade II-III) have fewer malignant characteristics than high-grade gliomas (WHO Grade IV), and a better clinical prognosis, however, accurate discrimination of overall survival (OS) remains a challenge. In this study, we aimed to identify tissue-derived image features using a machine learning approach to predict OS in a mixed histology and grade cohort of lower grade glioma patients. To achieve this aim, we used H and E stained slides from the public LGG cohort of The Cancer Genome Atlas (TCGA) to create a machine learned dictionary of "image-derived visual words" associated with OS. We then evaluated the combined efficacy of using these visual words in predicting short versus long OS by training a generalized machine learning model. Finally, we mapped these predictive visual words back to molecular signaling cascades to infer potential drivers of the machine learned survival-associated phenotypes. We analyzed digitized histological sections downloaded from the LGG cohort of TCGA using a bag-of-words approach. This method identified a diverse set of histological patterns that were further correlated with OS, histology, and molecular signaling activity using Cox regression, analysis of variance, and Spearman correlation, respectively. A support vector machine (SVM) model was constructed to discriminate patients into short and long OS groups dichotomized at 24-month. This method identified disease-relevant phenotypes associated with OS, some of which are correlated with disease-associated molecular pathways. From these image-derived phenotypes, a generalized SVM model which could

  8. [Histochemical stains for minerals by hematoxylin-lake method].

    Science.gov (United States)

    Miyagawa, Makoto

    2013-04-01

    The present study was undertaken to establish the experimental animal model by histological staining methods for minerals. After intraperitoneal injections of minerals, precipitates deposited on the surface of the liver. Liver tissues were fixed in paraformaldehyde, embedded in paraffin and cut into thin sections which were used as minerals containing standard section. Several reagents for histological stains and spectrophotometry for minerals were applied in both test-tube experiments and stainings of tissue sections to test for minerals. Hematoxylin-lake was found of capable of staining minerals in tissue. A simple technique used was described for light microscopic detection of minerals.

  9. Superresolution Imaging of Aquaporin-4 Cluster Size in Antibody-Stained Paraffin Brain Sections.

    Science.gov (United States)

    Smith, Alex J; Verkman, Alan S

    2015-12-15

    The water channel aquaporin-4 (AQP4) forms supramolecular clusters whose size is determined by the ratio of M1- and M23-AQP4 isoforms. In cultured astrocytes, differences in the subcellular localization and macromolecular interactions of small and large AQP4 clusters results in distinct physiological roles for M1- and M23-AQP4. Here, we developed quantitative superresolution optical imaging methodology to measure AQP4 cluster size in antibody-stained paraffin sections of mouse cerebral cortex and spinal cord, human postmortem brain, and glioma biopsy specimens. This methodology was used to demonstrate that large AQP4 clusters are formed in AQP4(-/-) astrocytes transfected with only M23-AQP4, but not in those expressing only M1-AQP4, both in vitro and in vivo. Native AQP4 in mouse cortex, where both isoforms are expressed, was enriched in astrocyte foot-processes adjacent to microcapillaries; clusters in perivascular regions of the cortex were larger than in parenchymal regions, demonstrating size-dependent subcellular segregation of AQP4 clusters. Two-color superresolution imaging demonstrated colocalization of Kir4.1 with AQP4 clusters in perivascular areas but not in parenchyma. Surprisingly, the subcellular distribution of AQP4 clusters was different between gray and white matter astrocytes in spinal cord, demonstrating regional specificity in cluster polarization. Changes in AQP4 subcellular distribution are associated with several neurological diseases and we demonstrate that AQP4 clustering was preserved in a postmortem human cortical brain tissue specimen, but that AQP4 was not substantially clustered in a human glioblastoma specimen despite high-level expression. Our results demonstrate the utility of superresolution optical imaging for measuring the size of AQP4 supramolecular clusters in paraffin sections of brain tissue and support AQP4 cluster size as a primary determinant of its subcellular distribution. Copyright © 2015 Biophysical Society

  10. Endoanal MRI of the anal sphincter complex: correlation with cross-sectional anatomy and histology

    NARCIS (Netherlands)

    Hussain, S. M.; Stoker, J.; Zwamborn, A. W.; den Hollander, J. C.; Kuiper, J. W.; Entius, C. A.; Laméris, J. S.

    1996-01-01

    The purpose of this study was to correlate the in vivo endoanal MRI findings of the anal sphincter with the cross-sectional anatomy and histology. Fourteen patients with rectal tumours were examined with a rigid endoanal MR coil before undergoing abdominoperineal resection. In addition, 12 cadavers

  11. Endoanal MRI of the anal sphincter complex: correlation with cross-sectional anatomy and histology

    NARCIS (Netherlands)

    S.M. Hussain (Shahid); J. Stoker (Jacob); A.W. Zwamborn; J.C. den Hollander (Jan); J.-W. Kuiper (Jan-Willem); C.A. Entius; J.S. Lameris

    1996-01-01

    textabstractThe purpose of this study was to correlate the in vivo endoanal MRI findings of the anal sphincter with the cross-sectional anatomy and histology. Fourteen patients with rectal tumours were examined with a rigid endoanal MR coil before undergoing

  12. Correlation between ultrasound imaging, cross-sectional anatomy, and histology of the brachial plexus: a review.

    Science.gov (United States)

    van Geffen, Geert J; Moayeri, Nizar; Bruhn, Jörgen; Scheffer, Gert J; Chan, Vincent W; Groen, Gerbrand J

    2009-01-01

    The anatomy of the brachial plexus is complex. To facilitate the understanding of the ultrasound appearance of the brachial plexus, we present a review of important anatomic considerations. A detailed correlation of reconstructed, cross-sectional gross anatomy and histology with ultrasound sonoanatomy is provided.

  13. Improved method of producing satisfactory sections of whole eyeball by routine histology.

    Science.gov (United States)

    Arko-Boham, Benjamin; Ahenkorah, John; Hottor, Bismarck Afedo; Dennis, Esther; Addai, Frederick Kwaku

    2014-02-01

    To overcome the loss of structural integrity when eyeball sections are prepared by wax embedding, we experimentally modified the routine histological procedure and report satisfactorily well-preserved antero-posterior sections of whole eyeballs for teaching/learning purposes. Presently histological sections of whole eyeballs are not readily available because substantial structural distortions attributable to variable consistency of tissue components (and their undesired differential shrinkage) result from routine processing. Notably, at the dehydration stage of processing, the soft, gel-like vitreous humor considerably shrinks relative to the tough fibrous sclera causing collapse of the ocular globe. Additionally, the combined effects of fixation, dehydration, and embedding at 60°C renders the eye lens too hard for microtome slicing at thicknesses suitable for light microscopy. We satisfactorily preserved intact antero-posterior sections of eyeballs via routine paraffin wax processing procedure entailing two main modifications; (i) careful needle aspiration of vitreous humor and replacement with molten wax prior to wax infiltration; (ii) softening of lens in trimmed wax block by placing a drop of concentrated liquid phenol on it for 3 h during microtomy. These variations of the routine histological method produced intact whole eyeball sections with retinal detachment as the only structural distortion. Intact sections of the eyeball obtained compares well with the laborious, expensive, and 8-week long celloidin method. Our method has wider potential usability than costly freeze drying method which requires special skills and equipment (cryotome) and does not produce whole eyeball sections. Copyright © 2013 Wiley Periodicals, Inc.

  14. Diagnostic utility of melanin production by fungi: Study on tissue sections and culture smears with Masson-Fontana stain

    Directory of Open Access Journals (Sweden)

    Challa Sundaram

    2014-01-01

    Full Text Available Background: Dematiaceous fungi appear brown in tissue section due to melanin in their cell walls. When the brown color is not seen on routine H and E and culture is not available, differentiation of dematiaceous fungi from other fungi is difficult on morphology alone. Aims and Objective: To study if melanin production by dematiaceous fungi can help differentiate them from other types of fungi. Materials and Methods: Fifty tissue sections of various fungal infections and 13 smears from cultures of different species of fungi were stained with Masson Fontana stain to assess melanin production. The tissue sections included biopsies from 26 culture-proven fungi and 24 biopsies of filamentous fungi diagnosed on morphology alone with no culture confirmation. Results: All culture-proven dematiaceous fungi and Zygomycetes showed strong positivity in sections and culture smears. Aspergillus sp showed variable positivity and intensity. Cryptococcus neoformans showed strong positivity in tissue sections and culture smears. Tissue sections of septate filamentous fungi (9/15, Zygomycetes (4/5, and fungi with both hyphal and yeast morphology (4/4 showed positivity for melanin. The septate filamentous fungi negative for melanin were from biopsy samples of fungal sinusitis including both allergic and invasive fungal sinusitis and colonizing fungal balls. Conclusion: Melanin is produced by both dematiaceous and non-dematiaceous fungi. Masson-Fontana stain cannot reliably differentiate dematiaceous fungi from other filamentous fungi like Aspergillus sp; however, absence of melanin in the hyphae may be used to rule out dematiaceous fungi from other filamentous fungi. In the differential diagnosis of yeast fungi, Cryptococcus sp can be differentiated from Candida sp by Masson-Fontana stain in tissue sections.

  15. Double Pass 595 nm Pulsed Dye Laser Does Not Enhance the Efficacy of Port Wine Stains Compared with Single Pass: A Randomized Comparison with Histological Examination.

    Science.gov (United States)

    Yu, Wenxin; Zhu, Jiafang; Wang, Lizhen; Qiu, Yajing; Chen, Yijie; Yang, Xi; Chang, Lei; Ma, Gang; Lin, Xiaoxi

    2018-03-27

    To compare the efficacy and safety of double-pass pulsed dye laser (DWL) and single-pass PDL (SWL) in treating virgin port wine stain (PWS). The increase in the extent of vascular damage attributed to the use of double-pass techniques for PWS remains inconclusive. A prospective, side-by-side comparison with a histological study for virgin PWS is still lacking. Twenty-one patients (11 flat PWS, 10 hypertrophic PWS) with untreated PWS underwent 3 treatments at 2-month intervals. Each PWS was divided into three treatment sites: SWL, DWL, and untreated control. Chromametric and visual evaluation of the efficacy and evaluation of side effects were conducted 3 months after final treatment. Biopsies were taken at the treated sites immediately posttreatment. Chromametric and visual evaluation suggested that DWL sites showed no significant improvement compared with SWL (p > 0.05) in treating PWS. The mean depth of photothermal damage to the vessels was limited to a maximum of 0.36-0.41 mm in both SWL and DWL sides. Permanent side effects were not observed in any patients. Double-pass PDL does not enhance PWS clearance. To improve the clearance of PWS lesions, either the depth of laser penetration should be increased or greater photothermal damage to vessels should be generated.

  16. Correlation among ultrasound, cross-sectional anatomy, and histology of the sciatic nerve: a review.

    Science.gov (United States)

    Moayeri, Nizar; van Geffen, Geert J; Bruhn, Jörgen; Chan, Vincent W; Groen, Gerbrand J

    2010-01-01

    Efficient identification of the sciatic nerve (SN) requires a thorough knowledge of its topography in relation to the surrounding structures. Anatomic cross sections in similar oblique planes as observed during SN ultrasonography are lacking. A survey of sonoanatomy matched with ultrasound views of the major SN block sites will be helpful in pattern recognition, especially when combined with images that show the internal architecture of the nerve. From 1 cadaver, consecutive parts of the upper leg corresponding to the 4 major blocks sites were sectioned and deeply frozen. Using cryomicrotomy, consecutive transverse sections were acquired and photographed at 78-microm intervals, along with histologic sections at 5-mm intervals. Multiplanar reformatting was done to reconstruct the optimal planes for an accurate comparison of ultrasonography and gross anatomy. The anatomic and histologic images were matched with ultrasound images that were obtained from 2 healthy volunteers. By simulating the exact position and angulation as in the ultrasonographic images, detailed anatomic overviews of SN and adjacent structures were reconstructed in the gluteal, subgluteal, midfemoral, and popliteal regions. Throughout its trajectory, SN contains numerous fascicles with connective and adipose tissues. In this study, we provide an optimal matching between histology, anatomic cross sections, and short-axis ultrasound images of SN. Reconstructing ultrasonographic planes with this high-resolution digitized anatomy not only enables an overview but also shows detailed views of the architecture of internal SN. The undulating course of the nerve fascicles within SN may explain its varying echogenic appearance during probe manipulation.

  17. Slice-to-Volume Nonrigid Registration of Histological Sections to MR Images of the Human Brain

    Science.gov (United States)

    Osechinskiy, Sergey; Kruggel, Frithjof

    2011-01-01

    Registration of histological images to three-dimensional imaging modalities is an important step in quantitative analysis of brain structure, in architectonic mapping of the brain, and in investigation of the pathology of a brain disease. Reconstruction of histology volume from serial sections is a well-established procedure, but it does not address registration of individual slices from sparse sections, which is the aim of the slice-to-volume approach. This study presents a flexible framework for intensity-based slice-to-volume nonrigid registration algorithms with a geometric transformation deformation field parametrized by various classes of spline functions: thin-plate splines (TPS), Gaussian elastic body splines (GEBS), or cubic B-splines. Algorithms are applied to cross-modality registration of histological and magnetic resonance images of the human brain. Registration performance is evaluated across a range of optimization algorithms and intensity-based cost functions. For a particular case of histological data, best results are obtained with a TPS three-dimensional (3D) warp, a new unconstrained optimization algorithm (NEWUOA), and a correlation-coefficient-based cost function. PMID:22567290

  18. Slice-to-Volume Nonrigid Registration of Histological Sections to MR Images of the Human Brain

    Directory of Open Access Journals (Sweden)

    Sergey Osechinskiy

    2011-01-01

    Full Text Available Registration of histological images to three-dimensional imaging modalities is an important step in quantitative analysis of brain structure, in architectonic mapping of the brain, and in investigation of the pathology of a brain disease. Reconstruction of histology volume from serial sections is a well-established procedure, but it does not address registration of individual slices from sparse sections, which is the aim of the slice-to-volume approach. This study presents a flexible framework for intensity-based slice-to-volume nonrigid registration algorithms with a geometric transformation deformation field parametrized by various classes of spline functions: thin-plate splines (TPS, Gaussian elastic body splines (GEBS, or cubic B-splines. Algorithms are applied to cross-modality registration of histological and magnetic resonance images of the human brain. Registration performance is evaluated across a range of optimization algorithms and intensity-based cost functions. For a particular case of histological data, best results are obtained with a TPS three-dimensional (3D warp, a new unconstrained optimization algorithm (NEWUOA, and a correlation-coefficient-based cost function.

  19. [Application of Immunohistochemistry and Immunofluorescence Staining in Detection of Phospholipase A2 Receptor on Paraffin Section of Renal Biopsy Tissue].

    Science.gov (United States)

    Dong, Hong-rui; Wang, Yan-yan; Wang, Guo-qin; Sun, Li-jun; Cheng, Hong; Chen, Yi-pu

    2015-10-01

    To evaluate the application of immunohistochemistry and fluorescence staining method in the detection of phospholipase A2 receptor (PLA2R) on paraffin section of renal biopsy tissue,and to find an accurate and fast method for the detection of PLA2R in renal tissue. The PLA2R of 193 cases were detected by immunohistochemical staining,and the antigen was repaired by the method of high pressure cooker (HPC) hot repair plus trypsin repair. The 193 samples including 139 cases of idiopathic membranous nephropathy (IMN), 15 cases of membranous lupus nephritis, 8 cases of hepatitis B virus associated membranous nephropathy, 18 cases of IgA nephropathy, and 13 cases of minimal change diseases. To compare the dyeing effects, 22 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 4 different. of antigen repairing,which included HPC hot repair, HPC hot repair plus trypsin repair, water bath heat repair, and water bath heat repair plus trypsin repair. To compare the dyeing effects, 15 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 3 different. of antigen repairing,which included water bath heat repair plus trypsin repair, protease K digestion repair, and pepsin digestion repair. In 193 cases, the positive rate of PLA2R in IMN cases was 90.6% (126/139), and the other 54 patients without IMN were negative. Twenty-two IMN patients were positive for PLA2R by using the HPC heat repair plus trypsin repaire or the water bath heat repair plus trypsin repair;while only a few cases of 22 IMN cases were positive by using the HPC hot repair alone or water bath heat repair alone. Fifteen IMN patients were positive for PLA2R by using water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair, but the distribution of positive deposits and the background were different. PLA2R immunohistochemical staining can effectively identify IMN and secondary MN. For

  20. Factors influencing extract of Hibiscus sabdariffa staining of rat testes.

    Science.gov (United States)

    Bassey, R B; Bakare, A A; Peter, A I; Oremosu, A A; Osinubi, A A

    2012-08-01

    Some plant extracts can be used in biology and medicine to reveal or identify cellular components and tissues. We investigated the effects of time and concentration on staining of histological sections of rat testes by an acidified extract of Hibiscus sabdariffa. An ethanolic extract of H. sabdariffa was diluted using 1% acetic acid in 70% ethanol to stain histological sections of testes at concentrations of 0.2, 0.1 and 0.05 g/ml for 5, 10, 15, 30, 45 and 60 min. The sections of testes were stained deep red. The staining efficiency of H. sabdariffa was greater at a high concentration and required less time to achieve optimal staining. H. sabdariffa is a strongly basic dye that can be used for various diagnostic purposes. Staining time and concentration must be considered to achieve optimal results.

  1. Atomic force microscopy of histological sections using a chemical etching method

    International Nuclear Information System (INIS)

    Tiribilli, B.; Bani, D.; Quercioli, F.; Ghirelli, A.; Vassalli, M.

    2005-01-01

    Physiology and pathology have a big deal on tissue morphology, and the intrinsic spatial resolution of an atomic force microscope (AFM) is able to observe ultrastructural details. In order to investigate cellular and subcellular structures in histological sections with the AFM, we used a new simple method for sample preparation, i.e. chemical etching of semithin sections from epoxy resin-embedded specimens: such treatment appears to melt the upper layers of the embedding resin; thus, removing the superficial roughness caused by the edge of the microtome knife and bringing into high relief the biological structures hidden in the bulk. Consecutive ultrathin sections embedded in epoxy resin were observed with a transmission electron microscope (TEM) to compare the different imaging properties on the same specimen sample. In this paper we report, as an example, our AFM and TEM images of two different tissue specimens, rat pancreas and skeletal muscle fibres, showing that most of the inner details are visible with the AFM. These results suggest that chemical etching of histological sections may be a simple, fast and cost-effective method for AFM imaging with ultrastructural resolution

  2. Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning

    Science.gov (United States)

    Luciano, Nicholas J; Sati, Pascal; Nair, Govind; Guy, Joseph R; Ha, Seung-Kwon; Absinta, Martina; Chiang, Wen-Yang; Leibovitch, Emily C; Jacobson, Steven; Silva, Afonso C; Reich, Daniel S.

    2016-01-01

    Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume three-dimensionally. While MRI is an extremely sensitive tool for detecting tissue abnormalities, association of signal changes with an underlying pathological process is usually not straightforward. In the central nervous system, for example, inflammation, demyelination, axonal damage, gliosis, and neuronal death may all induce similar findings on MRI. As such, interpretation of MRI scans depends on the context, and radiological-histopathological correlation is therefore of the utmost importance. Unfortunately, traditional pathological sectioning of brain tissue is often imprecise and inconsistent, thus complicating the comparison between histology sections and MRI. This article presents novel methodology for accurately sectioning primate brain tissues and thus allowing precise matching between histology and MRI. The detailed protocol described in this article will assist investigators in applying this method, which relies on the creation of 3D printed brain slicers. Slightly modified, it can be easily implemented for brains of other species, including humans. PMID:28060281

  3. Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning.

    Science.gov (United States)

    Luciano, Nicholas J; Sati, Pascal; Nair, Govind; Guy, Joseph R; Ha, Seung-Kwon; Absinta, Martina; Chiang, Wen-Yang; Leibovitch, Emily C; Jacobson, Steven; Silva, Afonso C; Reich, Daniel S

    2016-12-06

    Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume three-dimensionally. While MRI is an extremely sensitive tool for detecting tissue abnormalities, association of signal changes with an underlying pathological process is usually not straightforward. In the central nervous system, for example, inflammation, demyelination, axonal damage, gliosis, and neuronal death may all induce similar findings on MRI. As such, interpretation of MRI scans depends on the context, and radiological-histopathological correlation is therefore of the utmost importance. Unfortunately, traditional pathological sectioning of brain tissue is often imprecise and inconsistent, thus complicating the comparison between histology sections and MRI. This article presents novel methodology for accurately sectioning primate brain tissues and thus allowing precise matching between histology and MRI. The detailed protocol described in this article will assist investigators in applying this method, which relies on the creation of 3D printed brain slicers. Slightly modified, it can be easily implemented for brains of other species, including humans.

  4. Automated image segmentation of haematoxylin and eosin stained skeletal muscle cross-sections

    DEFF Research Database (Denmark)

    Liu, F; Mackey, AL; Srikuea, R

    2013-01-01

    of two major steps: (1) A learning-based seed detection method to find the geometric centres of the muscle fibres, and (2) a colour gradient repulsive balloon snake deformable model that adopts colour gradient in Luv colour space. Automatic quantification of muscle fibre cross-sectional areas using...

  5. Human Chorioretinal Layer Thicknesses Measured in Macula-wide, High-Resolution Histologic Sections

    Science.gov (United States)

    Messinger, Jeffrey D.; Sloan, Kenneth R.; Mitra, Arnab; McGwin, Gerald; Spaide, Richard F.

    2011-01-01

    Purpose. To provide a comprehensive description of chorioretinal layer thicknesses in the normal human macula, including two-layer pairs that can produce a combined signal in some optical coherence tomography (OCT) devices (ganglion cell [GCL] and inner plexiform [IPL] layers and outer plexiform [OPL] and outer nuclear [ONL] layers). Methods. In 0.8-μm-thick, macula-wide sections through the foveola of 18 donors (age range, 40–92 years), 21 layers were measured at 25 locations by a trained observer and validated by a second observer. Tissue volume changes were assessed by comparing total retinal thickness in ex vivo OCT and in sections. Results. Median tissue shrinkage was 14.5% overall and 29% in the fovea. Histologic laminar boundaries resembled those in SD-OCT scans, but the shapes of the foveolar OPL and ONL differed. Histologic GCL, IPL, and OPLHenle were thickest at 0.8. to 1, 1.5, and 0.4 mm eccentricity, respectively. ONL was thickest in an inward bulge at the foveal center. At 1 mm eccentricity, GCL, INL, and OPLHenle represented 17.3% to 21.1%, 18.0% to 18.5%, and 14.2% to 16.6% of total retinal thickness, respectively. In donors ≥70 years of age, the RPE and choroid were 17.1% and 29.6% thinner and OPLHenle was 20.8% thicker than in donors macula were generated. Newer OCT systems can separate GCL from IPL and OPLHenle from ONL, with good agreement for the proportion of retinal thickness occupied by OPLHenle in OCT and histology. The thickening of OPLHenle in older eyes may reflect Müller cell hypertrophy associated with rod loss. PMID:21421869

  6. Multi-Resolution Wavelet-Transformed Image Analysis of Histological Sections of Breast Carcinomas

    Directory of Open Access Journals (Sweden)

    Hae-Gil Hwang

    2005-01-01

    Full Text Available Multi-resolution images of histological sections of breast cancer tissue were analyzed using texture features of Haar- and Daubechies transform wavelets. Tissue samples analyzed were from ductal regions of the breast and included benign ductal hyperplasia, ductal carcinoma in situ (DCIS, and invasive ductal carcinoma (CA. To assess the correlation between computerized image analysis and visual analysis by a pathologist, we created a two-step classification system based on feature extraction and classification. In the feature extraction step, we extracted texture features from wavelet-transformed images at 10× magnification. In the classification step, we applied two types of classifiers to the extracted features, namely a statistics-based multivariate (discriminant analysis and a neural network. Using features from second-level Haar transform wavelet images in combination with discriminant analysis, we obtained classification accuracies of 96.67 and 87.78% for the training and testing set (90 images each, respectively. We conclude that the best classifier of carcinomas in histological sections of breast tissue are the texture features from the second-level Haar transform wavelet images used in a discriminant function.

  7. Large deformation diffeomorphic metric mapping registration of reconstructed 3D histological section images and in vivo MR images

    Directory of Open Access Journals (Sweden)

    Can Ceritoglu

    2010-05-01

    Full Text Available Our current understanding of neuroanatomical abnormalities in neuropsychiatric diseases is based largely on magnetic resonance imaging (MRI and post mortem histological analyses of the brain. Further advances in elucidating altered brain structure in these human conditions might emerge from combining MRI and histological methods. We propose a multistage method for registering 3D volumes reconstructed from histological sections to corresponding in vivo MRI volumes from the same subjects: (1 manual segmentation of white matter (WM, gray matter (GM and cerebrospinal fluid (CSF compartments in histological sections, (2 alignment of consecutive histological sections using 2D rigid transformation to construct a 3D histological image volume from the aligned sections, (3 registration of reconstructed 3D histological volumes to the corresponding 3D MRI volumes using 3D affine transformation, (4 intensity normalization of images via histogram matching and (5 registration of the volumes via intensity based Large Deformation Diffeomorphic Metric (LDDMM image matching algorithm. Here we demonstrate the utility of our method in the transfer of cytoarchitectonic information from histological sections to identify regions of interest in MRI scans of nine adult macaque brains for morphometric analyses. LDDMM improved the accuracy of the registration via decreased distances between GM/CSF surfaces after LDDMM (0.39±0.13 mm compared to distances after affine registration (0.76±0.41 mm. Similarly, WM/GM distances decreased to 0.28±0.16 mm after LDDMM compared to 0.54±0.39 mm after affine registration. The multistage registration method may find broad application for mapping histologically based information, e.g., receptor distributions, gene expression, onto MRI volumes.

  8. Histologic evaluation of skin damage after overlapping and nonoverlapping flashlamp pumped pulsed dye laser pulses: A study on normal human skin as a model for port wine stains

    NARCIS (Netherlands)

    Koster, P. H.; van der Horst, C. M.; van Gemert, M. J.; van der Wal, A. C.

    2001-01-01

    BACKGROUND AND OBJECTIVE: In the treatment of port wine stains (PWS) with the flashlamp pumped pulsed dye laser (FPPDL), no consensus exists about overlapping of pulses. The advantage of overlapping pulses is homogeneous lightening of the PWS; the risk is redundant tissue damage. The aim of this

  9. Evaluating a technology supported interactive response system during the laboratory section of a histology course.

    Science.gov (United States)

    Rinaldi, Vera D; Lorr, Nancy A; Williams, Kimberly

    2017-07-01

    Monitoring of student learning through systematic formative assessment is important for adjusting pedagogical strategies. However, traditional formative assessments, such as quizzes and written assignments, may not be sufficiently timely for making adjustments to a learning process. Technology supported formative assessment tools assess student knowledge, allow for immediate feedback, facilitate classroom dialogues, and have the potential to modify student learning strategies. As an attempt to integrate technology supported formative assessment in the laboratory section of an upper-level histology course, the interactive application Learning Catalytics TM , a cloud-based assessment system, was used. This study conducted during the 2015 Histology courses at Cornell University concluded that this application is helpful for identifying student misconceptions "on-the-go," engaging otherwise marginalized students, and forming a new communication venue between students and instructors. There was no overall difference between grades from topics that used the application and grades from those that did not, and students reported that it only slightly helped improve their understanding of the topic (3.8 ± 0.99 on a five-point Likert scale). However, they highly recommended using it (4.2 ± 0.71). The major limitation was regarding the image display and graphical resolution of this application. Even though students embrace the use of technology, 39% reported benefits of having the traditional light microscope available. This cohort of students led instructors to conclude that the newest tools are not always better, but rather can complement traditional instruction methods. Anat Sci Educ 10: 328-338. © 2016 American Association of Anatomists. © 2016 American Association of Anatomists.

  10. HistoStitcher© : An Interactive Program for Accurate and Rapid Reconstruction of Digitized Whole Histological Sections from Tissue Fragments

    Science.gov (United States)

    Chappelow, Jonathan; Tomaszewski, John E.; Feldman, Michael; Shih, Natalie; Madabhushi, Anant

    2011-01-01

    We present an interactive program called HistoStitcher© for accurate and rapid reassembly of histology fragments into a pseudo-whole digitized histological section. HistoStitcher© provides both an intuitive graphical interface to assist the operator in performing the stitch of adjacent histology fragments by selecting pairs of anatomical landmarks, and a set of computational routines for determining and applying an optimal linear transformation to generate the stitched image. Reconstruction of whole histological sections from images of slides containing smaller fragments is required in applications where preparation of whole sections of large tissue specimens is not feasible or efficient, and such whole mounts are required to facilitate (a) disease annotation and (b) image registration with radiological images. Unlike manual reassembly of image fragments in a general purpose image editing program (such as Photoshop), HistoStitcher© provides memory efficient operation on high resolution digitized histology images and a highly flexible stitching process capable of producing more accurate results in less time. Further, by parameterizing the series of transformations determined by the stitching process, the stitching parameters can be saved, loaded at a later time, refined, or reapplied to multi-resolution scans, or quickly transmitted to another site. In this paper, we describe in detail the design of HistoStitcher© and the mathematical routines used for calculating the optimal image transformation, and demonstrate its operation for stitching high resolution histology quadrants of a prostate specimen to form a digitally reassembled whole histology section, for 8 different patient studies. To evaluate stitching quality, a 6 point scoring scheme, which assesses the alignment and continuity of anatomical structures important for disease annotation, is employed by three independent expert pathologists. For 6 studies compared with this scheme, reconstructed sections

  11. Combination therapy of radiation and immunomodulators in the treatment of MM46 tumor transplanted in C3H/He mice. Histological and histoenzymatic studies using alpha-naphtyl acetate esterase staining

    Energy Technology Data Exchange (ETDEWEB)

    Miyaji, Chihiro; Imanaka, Kazufumi; Gose, Kyuhei; Ichiyanagi, Akihiro; Imajo, Yoshinari (Kobe Univ. (Japan). School of Medicine)

    1983-12-01

    Female C3H/He mice aged 13 weeks with transplanted MM 46 tumor were used in histological and enzymohistochemical studies on the timing of administration of immunomodulators, PSK (a protein bound polysaccharide prepared from Coriolus versicolor), or OK-432 (Streptococcal preparation) combined with two fractionated local irradiation with the total dose of 3,000 rad. The daily dose of 250 mg/kg of PSK, or 1 KE/mouse of OK-432 was respectively injected intraperitoneally for four consecutive days before or after irradiation. Mice were sacrificed before irradiation and 7, 14 and 21 days after irradiation. Resected tumor tissues were examined using H-E staining and ..cap alpha..-naphtyl acetate esterase (ANAE) staining to observe the stromal reactions such as the infiltration of mononuclear cells and fibroblasts around tumor tissues. When PSK or OK-432 was administered after irradiation, remarkable lymphocytic infiltration was observed compared with the control group and the group to which PSK or OK-432 was administered before irradiation. ANAE staining revealed most of infiltrating lymphocytes were T-cells. These results suggested that PSK and OK-432 which were received after radiotherapy enhanced the immunological responses against tumors which were represented by remarkable lymphocytic infiltration.

  12. Objective automated quantification of fluorescence signal in histological sections of rat lens.

    Science.gov (United States)

    Talebizadeh, Nooshin; Hagström, Nanna Zhou; Yu, Zhaohua; Kronschläger, Martin; Söderberg, Per; Wählby, Carolina

    2017-08-01

    Visual quantification and classification of fluorescent signals is the gold standard in microscopy. The purpose of this study was to develop an automated method to delineate cells and to quantify expression of fluorescent signal of biomarkers in each nucleus and cytoplasm of lens epithelial cells in a histological section. A region of interest representing the lens epithelium was manually demarcated in each input image. Thereafter, individual cell nuclei within the region of interest were automatically delineated based on watershed segmentation and thresholding with an algorithm developed in Matlab™. Fluorescence signal was quantified within nuclei, cytoplasms and juxtaposed backgrounds. The classification of cells as labelled or not labelled was based on comparison of the fluorescence signal within cells with local background. The classification rule was thereafter optimized as compared with visual classification of a limited dataset. The performance of the automated classification was evaluated by asking 11 independent blinded observers to classify all cells (n = 395) in one lens image. Time consumed by the automatic algorithm and visual classification of cells was recorded. On an average, 77% of the cells were correctly classified as compared with the majority vote of the visual observers. The average agreement among visual observers was 83%. However, variation among visual observers was high, and agreement between two visual observers was as low as 71% in the worst case. Automated classification was on average 10 times faster than visual scoring. The presented method enables objective and fast detection of lens epithelial cells and quantification of expression of fluorescent signal with an accuracy comparable with the variability among visual observers. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  13. Is histologic esophagitis associated with dental erosion: a cross-sectional observational study?

    Science.gov (United States)

    Friesen, Lynn Roosa; Bohaty, Brenda; Onikul, Robin; Walker, Mary P; Abraham, Caren; Williams, Karen B; Cocjin, Jose T; Cocjin, Eileen L; Friesen, Craig A

    2017-08-10

    Gastroesophageal reflux disease (GERD) affects 15-25% of children and adolescents in the United States. The diagnosis of GERD in children is complex as reported symptoms or symptom profiles have been found to be unreliable. Frequently, the diagnosis must be confirmed by objective tests such as pH monitoring or histological evidence of esophagitis on an esophageal biopsy. Dental erosion has been shown to be associated with GERD as an atypical complication and has the potential to be a marker of GERD. The purposes of this study were to compare the frequency and patterns of dental erosion in children and adolescents with and without histologic esophagitis. Twenty-five subjects were recruited from patients scheduled for an upper gastrointestinal endoscopy. Information regarding potential GERD symptoms, food habits, and dental hygiene habits were obtained. Intra-oral photographs were taken, and a dental exam for erosion was performed. The results of a standard biopsy taken from the lower third of the esophagus during an endoscopy were used to divide subjects into either the control group or the GERD group (i.e. those with histologic esophagitis). Twenty-two subjects yielded 586 evaluable teeth. No significant difference was found between frequency or erosion patterns of those with and without histologic esophagitis. Dental erosions were more frequent in primary teeth. Dental erosions do not appear to be associated with histologic esophagitis indicative of GERD.

  14. Fluorescence excitation analysis by two-photon confocal laser scanning microscopy: a new method to identify fluorescent nanoparticles on histological tissue sections

    Directory of Open Access Journals (Sweden)

    Kahn E

    2012-10-01

    Full Text Available Edmond Kahn,1 Nicolas Tissot,3 Perrine Frere,3 Aurélien Dauphin,3 Mohamed Boumhras,2,4 Claude-Marie Bachelet,3 Frédérique Frouin,1 Gérard Lizard21Institut National de la Santé et de la Recherche Médicale (INSERM U678/UMR-S UPMC, CHU Pitié-Salpêtrière, Paris, France; 2Equipe Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique EA7270, Faculté des Sciences Gabriel, Université de Bourgogne-INSERM Dijon, France; 3Plateforme d'Imagerie cellulaire, UPMC, Paris, France; 4Laboratory of Biochemistry and Neuroscience, Applied Toxicology Group, Faculty of Science and Technology, Settat, MoroccoAbstract: In the present study, we make use of the ability of two-photon confocal laser scanning microscopes (CLSMs equipped with tunable lasers to produce spectral excitation image sequences. Furthermore, unmixing, which is usually performed on emission image sequences, is performed on these excitation image sequences. We use factor analysis of medical image sequences (FAMIS, which produces factor images, to unmix spectral image sequences of stained structures in tissue sections to provide images of characterized stained cellular structures. This new approach is applied to histological tissue sections of mouse aorta containing labeled iron nanoparticles stained with Texas Red and counterstained with SYTO13, to obtain visual information about the accumulation of these nanoparticles in the arterial wall. The possible presence of Texas Red is determined using a two-photon CLSM associated with FAMIS via the excitation spectra. Texas Red and SYTO13 are thus differentiated, and corresponding factor images specify their possible presence and cellular localization. In conclusion, the designed protocol shows that sequences of images obtained by excitation in a two-photon CLSM enables characterization of Texas Red-stained nanoparticles and other markers. This methodology offers an alternative and complementary solution to the conventional use of emission

  15. ANIMATED SERIAL SECTIONS, A TEACHING AID FOR ORAL HISTOLOGY AND EMBRYOLOGY.

    Science.gov (United States)

    HAYDEN, JESS; AND OTHERS

    THE RELATIVE EFFECTIVENESS OF TWO TYPES OF VISUAL MEDIA FOR THE DEVELOPMENT OF A THREE-DIMENSIONAL CONCEPT OF A GIVEN ANATOMICAL REGION WAS INVESTIGATED. EXPERIMENTAL AND CONTROL GROUPS WERE RANDOMLY SELECTED FROM 119 FIRST-YEAR MEDICAL AND DENTAL STUDENTS IN AN HISTOLOGY CLASS. BOTH GROUPS ATTENDED THE SAME ONE-HOUR LECTURE. FOLLOWING THE LECTURE…

  16. Técnicas de coloração para detecção de Encephalitozoon cuniculi em cortes histológicos Stain techniques for detection of Encephalitozoon cuniculi in tissue sections

    Directory of Open Access Journals (Sweden)

    Maria Anete Lallo

    2010-11-01

    Full Text Available No presente trabalho, foram avaliadas diferentes técnicas de coloração aplicadas a cortes histológicos para a identificação de Encephalitozoon cuniculi. Foram utilizados fragmentos hepáticos de camundongos Balb-c, imunossuprimidos com ciclofosfamida e inoculados com esporos de Encephalitozoon cuniculi. Os cortes histológicos incluídos em parafina foram corados pelas técnicas de hematoxilina-eosina (H-E, tricrômica modificada, Gram-Chromotrope, Giemsa, Brown-Hopps, PAS, Ziehl-Neelsen e Grocott, e ainda pela técnica de calcoflúor. As colorações azul de toluidina-fucsina e azul de toluidina foram utilizadas para os cortes incluídos em resina plástica. As técnicas de Gram-Chromotrope, Brown-Hopps e Ziehl-Neelsen foram as que permitiram melhor visualização dos esporos para o diagnóstico de E. cuniculi em cortes histológicos incluídos em parafina, uma vez que possibilitaram a clara diferenciação dos esporos de outras estruturas teciduais. Nos tecidos incluídos em resina plástica, as colorações de azul de toluidina-fucsina e azul de toluidina também facilitaram o encontro do agente. Por outro lado, a técnica tricrômica apresentou grande variabilidade nos resultados, sendo, portanto, pouco indicada para o diagnóstico em tecido. As demais técnicas - H-E, calcoflúor, Grocott, Giemsa e PAS - não permitiram a fácil identificação do E. cuniculi.In this study, it was investigated different staining techniques applied to histological sections for identification of Encephalitozoon cuniculi. It was used liver fragments from Balb-c mice immunosuppressed with cyclophosphamide and inoculated with spores of Encephalitozoon cuniculi. The histological paraffin embedded sections were stained with hematoxylin-eosin (H-E, modified trichrome, Gram-Chromotrope, Giemsa, Brown-Hopps, PAS, Ziehl-Neelsen, and Grocott and also by the technique of calcofluor. Toluidine blue-fuchsin and toluidine blue stainings were used for the cuts

  17. Gram staining.

    Science.gov (United States)

    Coico, Richard

    2005-10-01

    Named after Hans Christian Gram who developed the method in 1884, the Gram stain allows one to distinguish between Gram-positive and Gram-negative bacteria on the basis of differential staining with a crystal violet-iodine complex and a safranin counterstain. The cell walls of Gram-positive organisms retain this complex after treatment with alcohol and appear purple, whereas gram-negative organisms decolorize following such treatment and appear pink. The method described here is useful for assessing bacterial contamination of tissue culture samples or for examining the Gram stain status and morphological features of bacteria isolated from mixed or isolated bacterial cultures.

  18. Changing prevalence patterns in endoscopic and histological diagnosis of gastritis? Data from a cross-sectional Central European multicentre study.

    Science.gov (United States)

    Wolf, Eva-Maria; Plieschnegger, Wolfgang; Geppert, Michael; Wigginghaus, Bernd; Höss, Gabriele M; Eherer, Andreas; Schneider, Nora I; Hauer, Almuthe; Rehak, Peter; Vieth, Michael; Langner, Cord

    2014-05-01

    Traditionally, Helicobacter infection is considered to be the most common cause of gastritis. In the cross-sectional Central European histoGERD trial, we assessed the prevalence of different types of gastritis, correlating histological and endoscopic diagnoses. A total of 1123 individuals participated in an observational multicentre study. Endoscopists classified individuals as positive or negative for gastritis and rendered the putative cause. Pathologists evaluated biopsy specimens based upon the Updated Sydney System. Histological diagnosis of gastritis was made in 639 (56.9%) participants. In all, 210 (18.7%) individuals were diagnosed with Helicobacter gastritis, 215 (19.1%) with post Helicobacter gastritis, 234 (20.8%) with reactive gastropathy, 26 (2.3%) with autoimmune gastritis, and 6 (0.5%) with focally enhanced gastritis related to Crohn's disease. In 46 out of 639 (7.2%) individuals diagnosed with gastritis, combinations of different histological subtypes were noted the most common being reactive gastropathy and post Helicobacter gastritis. Endoscopic diagnosis of gastritis was made in 534 (47.6%) individuals. Reactive gastropathy was more common than active Helicobacter gastritis, and the majority of cases attributable to Helicobacter infection were no longer ongoing, i.e. post Helicobacter gastritis. Agreement between histological and endoscopic diagnoses was better in reactive gastropathy than in Helicobacter gastritis. Copyright © 2014 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

  19. Influence of type 2 diabetes mellitus on liver histology among morbidly obese individuals. A cross-sectional study.

    Science.gov (United States)

    Cazzo, Everton; Jimenez, Laísa Simakawa; Gallo, Fábio de Felice; Pareja, José Carlos; Chaim, Elinton Adami

    2016-01-01

    Nonalcoholic fatty liver disease (NAFLD) has become a public health concern. It encompasses a wide spectrum of histological abnormalities and has close relationships with insulin resistance and type 2 diabetes mellitus (T2DM). This study sought to compare the histological alterations observed in morbidly obese individuals with and without T2DM who underwent Roux-en-Y gastric bypass. Cross-sectional study in a tertiary-level public hospital. This was a cross-sectional study on 197 individuals who underwent gastric bypass surgery between 2011 and 2013. NAFLD was assessed through liver biopsies. T2DM was diagnosed through the International Diabetes Federation criteria. Non-diabetics presented significantly more biopsies without any histological abnormalities, regarding steatosis (42.6% versus 25.5%; P = 0.0400), fibrosis (60.6% versus 36.2%; P = 0.0042) and steatohepatitis (27.3% versus 12.8%; P = 0.0495), while diabetics presented significantly higher frequency of moderate forms of steatosis (36.2% versus 20%; P = 0.0307) and fibrosis (23.4% versus 4%; P = 0.0002). T2DM was associated with more advanced forms of NAFLD within the population studied. NAFLD has previously been correlated with severe forms of heart disease. Screening for and early detecting of NAFLD in high-risk populations are important for avoiding further development of severe forms and the need for liver transplantation.

  20. The Development of a Virtual 3D Model of the Renal Corpuscle from Serial Histological Sections for E-Learning Environments

    Science.gov (United States)

    Roth, Jeremy A.; Wilson, Timothy D.; Sandig, Martin

    2015-01-01

    Histology is a core subject in the anatomical sciences where learners are challenged to interpret two-dimensional (2D) information (gained from histological sections) to extrapolate and understand the three-dimensional (3D) morphology of cells, tissues, and organs. In gross anatomical education 3D models and learning tools have been associated…

  1. Histological step sectioning of pelvic lymph nodes increases the number of identified lymph node metastases

    DEFF Research Database (Denmark)

    Engvad, Birte; Poulsen, Mads H; Staun, Pia W

    2014-01-01

    Pathological examinations of lymph nodes (LN) in prostate cancer patients are handled differently at various institutions. The objective of this study is to provide means to improve the guidelines by examining the impact of step sectioning on LN status in patients with intermediate and high...

  2. Utilization of Cell-Transfer Technique for Molecular Testing on Hematoxylin-Eosin-Stained Sections: A Viable Option for Small Biopsies That Lack Tumor Tissues in Paraffin Block.

    Science.gov (United States)

    Wu, Howard H; Jovonovich, Stephen M; Randolph, Melissa; Post, Kristin M; Sen, Joyashree D; Curless, Kendra; Cheng, Liang

    2016-12-01

    - In some instances the standard method of doing molecular testing from formalin-fixed, paraffin-embedded block is not possible because of limited tissue. Tumor cell-enriched cell-transfer technique has been proven useful for performing immunocytochemistry and molecular testing on cytologic smears. - To establish the cell-transfer technique as a viable option for isolating tumor cells from hematoxylin-eosin (H&E)-stained slides. - Molecular testing was performed by using the cell-transfer technique on 97 archived H&E-stained slides from a variety of different tumors. Results were compared to the conventional method of molecular testing. - Polymerase chain reaction-based molecular testing via the cell-transfer technique was successfully performed on 82 of 97 samples (85%). This included 39 of 47 cases for EGFR, 10 of 11 cases for BRAF, and 33 of 39 cases for KRAS mutations. Eighty-one of 82 cell-transfer technique samples (99%) showed agreement with previous standard method results, including 4 mutations and 35 wild-type alleles for EGFR, 4 mutations and 6 wild-type alleles for BRAF, and 11 mutations and 21 wild-type alleles for KRAS. There was only 1 discrepancy: a cell-transfer technique with a false-negative >KRAS result (wild type versus G12C). - Molecular testing performed on H&E-stained sections via cell-transfer technique is useful when tissue from cell blocks and small surgical biopsy samples is exhausted and the only available material for testing is on H&E-stained slides.

  3. Differential staining of bacteria: acid fast stain.

    Science.gov (United States)

    Reynolds, Jackie; Moyes, Rita B; Breakwell, Donald P

    2009-11-01

    Acid-fastness is an uncommon characteristic shared by the genera Mycobacterium (Section 10A) and Nocardia. Because of this feature, this stain is extremely helpful in identification of these bacteria. Although Gram positive, acid-fast bacteria do not take the crystal violet into the wall well, appearing very light purple rather than the deep purple of normal Gram-positive bacteria. (c) 2009 by John Wiley & Sons, Inc.

  4. Backscattered electron SEM imaging of resin sections from plant specimens: observation of histological to subcellular structure and CLEM.

    Science.gov (United States)

    Rizzo, N W; Duncan, K E; Bourett, T M; Howard, R J

    2016-08-01

    We have refined methods for biological specimen preparation and low-voltage backscattered electron imaging in the scanning electron microscope that allow for observation at continuous magnifications of ca. 130-70 000 X, and documentation of tissue and subcellular ultrastructure detail. The technique, based upon early work by Ogura & Hasegawa (1980), affords use of significantly larger sections from fixed and resin-embedded specimens than is possible with transmission electron microscopy while providing similar data. After microtomy, the sections, typically ca. 750 nm thick, were dried onto the surface of glass or silicon wafer and stained with heavy metals-the use of grids avoided. The glass/wafer support was then mounted onto standard scanning electron microscopy sample stubs, carbon-coated and imaged directly at an accelerating voltage of 5 kV, using either a yttrium aluminum garnet or ExB backscattered electron detector. Alternatively, the sections could be viewed first by light microscopy, for example to document signal from a fluorescent protein, and then by scanning electron microscopy to provide correlative light/electron microscope (CLEM) data. These methods provide unobstructed access to ultrastructure in the spatial context of a section ca. 7 × 10 mm in size, significantly larger than the typical 0.2 × 0.3 mm section used for conventional transmission electron microscopy imaging. Application of this approach was especially useful when the biology of interest was rare or difficult to find, e.g. a particular cell type, developmental stage, large organ, the interface between cells of interacting organisms, when contextual information within a large tissue was obligatory, or combinations of these factors. In addition, the methods were easily adapted for immunolocalizations. © 2015 The Author. Journal of Microscopy published by John Wiley & Sons, Ltd on behalf of the Royal Microscopical Society.

  5. Optical-sectioning microscopy of protoporphyrin IX fluorescence in human gliomas: standardization and quantitative comparison with histology

    Science.gov (United States)

    Wei, Linpeng; Chen, Ye; Yin, Chengbo; Borwege, Sabine; Sanai, Nader; Liu, Jonathan T. C.

    2017-04-01

    Systemic delivery of 5-aminolevulinic acid leads to enhanced fluorescence image contrast in many tumors due to the increased accumulation of protoporphyrin IX (PpIX), a fluorescent porphyrin that is associated with tumor burden and proliferation. The value of PpIX-guided resection of malignant gliomas has been demonstrated in prospective randomized clinical studies in which a twofold greater extent of resection and improved progression-free survival have been observed. In low-grade gliomas and at the diffuse infiltrative margins of all gliomas, PpIX fluorescence is often too weak to be detected with current low-resolution surgical microscopes that are used in operating rooms. However, it has been demonstrated that high-resolution optical-sectioning microscopes are capable of detecting the sparse and punctate accumulations of PpIX that are undetectable via conventional low-power surgical fluorescence microscopes. To standardize the performance of high-resolution optical-sectioning devices for future clinical use, we have developed an imaging phantom and methods to ensure that the imaging of PpIX-expressing brain tissues can be performed reproducibly. Ex vivo imaging studies with a dual-axis confocal microscope demonstrate that these methods enable the acquisition of images from unsectioned human brain tissues that quantitatively and consistently correlate with images of histologically processed tissue sections.

  6. Comparison of bone-implant contact and bone-implant volume between 2D-histological sections and 3D-SRµCT slices

    Directory of Open Access Journals (Sweden)

    R Bernhardt

    2012-04-01

    Full Text Available Histological imaging is still considered the gold standard for analysing bone formation around metallic implants. Generally, a limited number of histological sections per sample are used for the approximation of mean values of peri-implant bone formation. In this study we compared statistically the results of bone-implant contact (BIC and bone-implant volume (BIV obtained by histological sections, with those obtained by X-ray absorption images from synchrotron radiation micro-computed tomography (SRµCT using osseointegrated screw-shaped implants from a mini-pig study. Comparing the BIC results of 3-4 histological sections per implant sample with the appropriate 3-4 SRµCT slices showed a non-significant difference of 1.9 % (p = 0.703. The contact area assessed by the whole 3D information from the SRµCT measurement in comparison to the histomorphometric results showed a non-significant difference in BIC of 4.9 % (p = 0.171. The amount of the bone-implant volume in the histological sections and the appropriate SRµCT slices showed a non-significant difference by only 1.4 % (p = 0.736 and also remains non-significant with 2.6 % (p = 0.323 using the volumetric SRµCT information. We conclude that for a clinical evaluation of implant osseointegration with histological imaging at least 3-4 sections per sample are sufficient to represent the BIC or BIV for a sample. Due to the fact that in this study we have found a significant intra-sample variation in BIC of up to ± 35 % the selection of only one or two histological sections per sample may strongly influence the determined BIC.

  7. Quantitative Image Analysis of Epithelial and Stromal Area in Histological Sections of Colorectal Cancer: An Emerging Diagnostic Tool

    Directory of Open Access Journals (Sweden)

    R. Rogojanu

    2015-01-01

    Full Text Available In colorectal cancer (CRC, an increase in the stromal (S area with the reduction of the epithelial (E parts has been suggested as an indication of tumor progression. Therefore, an automated image method capable of discriminating E and S areas would allow an improved diagnosis. Immunofluorescence staining was performed on paraffin-embedded sections from colorectal tumors (16 samples from patients with liver metastasis and 18 without. Noncancerous tumor adjacent mucosa (n=5 and normal mucosa (n=4 were taken as controls. Epithelial cells were identified by an anti-keratin 8 (K8 antibody. Large tissue areas (5–63 mm2/slide including tumor center, tumor front, and adjacent mucosa were scanned using an automated microscopy system (TissueFAXS. With our newly developed algorithms, we showed that there is more K8-immunoreactive E in the tumor center than in tumor adjacent and normal mucosa. Comparing patients with and without metastasis, the E/S ratio decreased by 20% in the tumor center and by 40% at tumor front in metastatic samples. The reduction of E might be due to a more aggressive phenotype in metastasis patients. The novel software allowed a detailed morphometric analysis of cancer tissue compartments as tools for objective quantitative measurements, reduced analysis time, and increased reproducibility of the data.

  8. Improved correlation of histological data with DCE MRI parameter maps by 3D reconstruction, reslicing and parameterization of the histological images

    International Nuclear Information System (INIS)

    Kiessling, Fabian; Le-Huu, Martin; Semmler, Wolfhard; Kunert, Tobias; Thorn, Matthias; Meinzer, Hans-Peter; Vosseler, Silvia; Fusenig, Norbert E.; Schmidt, Kerstin; Hoffend, Johannes

    2005-01-01

    Due to poor correlation of slice thickness and orientation, verification of radiological methods with histology is difficult. Thus, a procedure for three-dimensional reconstruction, reslicing and parameterization of histological data was developed, enabling a proper correlation with radiological data. Two different subcutaneous tumors were examined by MR microangiography and DCE-MRI, the latter being post-processed using a pharmacokinetic two-compartment model. Subsequently, tumors were serially sectioned and vessels stained with immunofluorescence markers. A ray-tracing algorithm performed three-dimensional visualization of the histological data, allowing virtually reslicing to thicker sections analogous to MRI slice geometry. Thick slices were processed as parameter maps color coding the marker density in the depth of the slice. Histological 3D reconstructions displayed the diffuse angioarchitecture of malignant tumors. Resliced histological images enabled specification of high enhancing areas seen on MR microangiography as large single vessels or vessel assemblies. In orthogonally reconstructed histological slices, single vessels were delineated. ROI analysis showed significant correlation between histological parameter maps of vessel density and MR parameter maps (r=0.83, P=0.05). The 3D approach to histology improves correlation of histological and radiological data due to proper matching of slice geometry. This method can be used with any histological stain, thus enabling a multivariable correlation of non-invasive data and histology. (orig.)

  9. Modification in the assembly technique of histological sections for analysis of spatial distribution of boron by autoradiography

    International Nuclear Information System (INIS)

    Portu, A; Carpano, M; Dagrosa, A; Pozzi, E; Thorp, S; Curotto, P; Cabrini, R L; Saint Martin, G

    2012-01-01

    The Boron Neutron Capture Therapy (BNCT) is a modality for the treatment of cancer, based on the capture reaction 10 B(n,α) 7 Li. The emitted particles are highly transferred linear of energy and have a short range in tissue (10 μ). Therefore, if the boron is selectively accumulates in tumor cellulo, the damage will be limited to preserving normal cellulo. Thus, the knowledge of the location of 10 B in the different structures of biological tissues as tumor and surrounding tissue, is essential when considering BNCT treatment (Barth et al., 2005). Neutron autoradiography is one of the few methods that allow studying the distribution spatial of elements emitters in a material containing such. As part of BNCT, the first step in performing autoradiography involves placing a freeze tissue section on a nuclear track detector (SSNTD) (Wittig et al., 2008). For this purpose, tissue samples are fixed in N 2 (liq) when they are resected after infusion boronated compound. The sample-detector arrangement is irradiated with thermal neutrons and elements cast in the capture reaction zones produce latent damage SSNTD. Chemically attacking the detector, this latent trace level can be amplified by optical microscopy. Thus, the distribution of 10 B in biological samples can be evaluated, so that this technique is suitable for studying the uptake of boron compounds for the different histological structures. In our laboratory, we have developed neutron autoradiography and has been applied to the study of different biological models (Portu et al., 2011a). In particular, the study conducted by the micro-distribution 10 B in tumors from nude mice model of cutaneous melanomas injected with boronophenylalanine (BPA) (Carpano et al, 2010;. Portu et al, 2011b.). Still using means of support for the sample to be cut, as OCTTM, the lack of structure of necrotic areas of tumors such causes tearing of these regions in the cutting process, which prevents achieving adequate for analysis sections

  10. A robust, efficient and flexible method for staining myelinated axons in blocks of brain tissue.

    Science.gov (United States)

    Wahlsten, Douglas; Colbourne, Frederick; Pleus, Richard

    2003-03-15

    Previous studies have demonstrated the utility of the gold chloride method for en bloc staining of a bisected brain in mice and rats. The present study explores several variations in the method, assesses its reliability, and extends the limits of its application. We conclude that the method is very efficient, highly robust, sufficiently accurate for most purposes, and adaptable to many morphometric measures. We obtained acceptable staining of commissures in every brain, despite a wide variety of fixation methods. One-half could be stained 24 h after the brain was extracted and the other half could be stained months later. When staining failed because of an exhausted solution, the brain could be stained successfully in fresh solution. Relatively small changes were found in the sizes of commissures several weeks after initial fixation or staining. A half brain stained to reveal the mid-sagittal section could then be sectioned coronally and stained again in either gold chloride for myelin or cresyl violet for Nissl substance. Uncertainty, arising from pixelation of digitized images was far less than errors arising from human judgments about the histological limits of major commissures. Useful data for morphometric analysis were obtained by scanning the surface of a gold chloride stained block of brain with an inexpensive flatbed scanner.

  11. Automated gastric cancer diagnosis on H&E-stained sections; ltraining a classifier on a large scale with multiple instance machine learning

    Science.gov (United States)

    Cosatto, Eric; Laquerre, Pierre-Francois; Malon, Christopher; Graf, Hans-Peter; Saito, Akira; Kiyuna, Tomoharu; Marugame, Atsushi; Kamijo, Ken'ichi

    2013-03-01

    We present a system that detects cancer on slides of gastric tissue sections stained with hematoxylin and eosin (H&E). At its heart is a classi er trained using the semi-supervised multi-instance learning framework (MIL) where each tissue is represented by a set of regions-of-interest (ROI) and a single label. Such labels are readily obtained because pathologists diagnose each tissue independently as part of the normal clinical work ow. From a large dataset of over 26K gastric tissue sections from over 12K patients obtained from a clinical load spanning several months, we train a MIL classi er on a patient-level partition of the dataset (2/3 of the patients) and obtain a very high performance of 96% (AUC), tested on the remaining 1/3 never-seen before patients (over 8K tissues). We show this level of performance to match the more costly supervised approach where individual ROIs need to be labeled manually. The large amount of data used to train this system gives us con dence in its robustness and that it can be safely used in a clinical setting. We demonstrate how it can improve the clinical work ow when used for pre-screening or quality control. For pre-screening, the system can diagnose 47% of the tissues with a very low likelihood (cancers, thus halving the clinicians' caseload. For quality control, compared to random rechecking of 33% of the cases, the system achieves a three-fold increase in the likelihood of catching cancers missed by pathologists. The system is currently in regular use at independent pathology labs in Japan where it is used to double-check clinician's diagnoses. At the end of 2012 it will have analyzed over 80,000 slides of gastric and colorectal samples (200,000 tissues).

  12. Stereological measures of trabecular bone structure: comparison of 3D micro computed tomography with 2D histological sections in human proximal tibial bone biopsies

    DEFF Research Database (Denmark)

    Thomsen, Jesper Skovhus; Laib, A.; Koller, B.

    2005-01-01

    Stereology applied on histological sections is the 'gold standard' for obtaining quantitative information on cancellous bone structure. Recent advances in micro computed tomography (microCT) have made it possible to acquire three-dimensional (3D) data non-destructively. However, before the 3D...... methods can be used as a substitute for the current 'gold standard' they have to be verified against the existing standard. The aim of this study was to compare bone structural measures obtained from 3D microCT data sets with those obtained by stereology performed on conventional histological sections...... tibial metaphysis. The biopsies were embedded in methylmetacrylate before microCT scanning in a Scanco microCT 40 scanner at a resolution of 20 x 20 x 20 microm3, and the 3D data sets were analysed with a computer program. After microCT scanning, 16 sections were cut from the central 2 mm of each biopsy...

  13. Detection of amyloid in abdominal fat pad aspirates in early amyloidosis: Role of electron microscopy and Congo red stained cell block sections

    Directory of Open Access Journals (Sweden)

    Sumana Devata

    2011-01-01

    Full Text Available Background: Fine-needle aspiration biopsy (FNA of the abdominal fat pad is a minimally invasive procedure to demonstrate tissue deposits of amyloid. However, protocols to evaluate amyloid in fat pad aspirates are not standardized, especially for detecting scant amyloid in early disease. Materials and Methods: We studied abdominal fat pad aspirates from 33 randomly selected patients in whom subsequent tissue biopsy, autopsy, and/or medical history for confirmation of amyloidosis (AL were also available. All these cases were suspected to have early AL, but had negative results on abdominal fat pad aspirates evaluated by polarizing microscopy of Congo Red stained sections (CRPM. The results with CRPM between four reviewers were compared in 12 cases for studying inter observer reproducibility. 24 cases were also evaluated by ultrastructural study with electron microscopy (EM. Results: Nine of thirty-three (27% cases reported negative by polarizing microscopy had amyloidosis. Reanalysis of 12 mixed positive-negative cases, showed considerable inter-observer variability with frequent lack of agreement between four observers by CRPM alone (Cohen′s Kappa index of 0.1, 95% CI -0.1 to 0.36. EM showed amyloid in the walls of small blood vessels in fibroadipose tissue in four out of nine cases (44% with amyloidosis. Conclusion: In addition to poor inter-observer reproducibility, CRPM alone in cases with scant amyloid led to frequent false negative results (9 out of 9, 100%. For improved detection of AL, routine ultrastructural evaluation with EM of fat pad aspirates by evaluating at least 15 small blood vessels in the aspirated fibroadipose tissue is recommended. Given the high false negative rate for CRPM alone in early disease, routine reflex evaluation with EM is highly recommended to avert the invasive option of biopsying various organs in cases with high clinical suspicion for AL.

  14. A new method for evaluating tumor-infiltrating lymphocytes (TILs) in colorectal cancer using hematoxylin and eosin (H-E)-stained tumor sections.

    Science.gov (United States)

    Iseki, Yasuhito; Shibutani, Masatsune; Maeda, Kiyoshi; Nagahara, Hisashi; Fukuoka, Tatsunari; Matsutani, Shinji; Kashiwagi, Shinichiro; Tanaka, Hiroaki; Hirakawa, Kosei; Ohira, Masaichi

    2018-01-01

    Numerous reports indicate that tumor-infiltrating lymphocytes (TILs) are a prognostic factor in various cancers and that they must be good biomarkers. However, the methods of evaluating TILs differ in each study; thus, there is not yet a standardized methodology for evaluating TILs. The purpose of this study is to evaluate the prognostic significance of tumor-infiltrating lymphocytes (TILs) in patients with colorectal cancer (CRC) using the new method proposed by the International TILs Working Group in breast cancer and to standardize the method of evaluating TILs in CRC. We retrospectively reviewed a database of 160 patients with Stage II or III CRC. The density of TILs was assessed by measuring the area occupied by mononuclear cells over the stromal area on hematoxylin and eosin (H-E)-stained sections. We set 42% as the cut-off percentage of the area occupied by TILs according to the receiver operating characteristic curve, and we classified patients into the high-TILs and the low-TILs groups. The rates of relapse-free survival (RFS) and overall survival (OS) in the high-TILs group were significantly higher than those in the low-TILs group. A multivariate analysis showed that the density of TILs was independently associated with RFS and OS. Moreover, the density of TILs assessed by an observer was significantly associated with the density of TILs assessed by the automated imaging software program. The new method for evaluating TILs, which was recommended by the International TILs Working Group in breast cancer, might be a useful predictive factor in colorectal cancer patients.

  15. Comparison of histologic margin status in low-grade cutaneous and subcutaneous canine mast cell tumours examined by radial and tangential sections.

    Science.gov (United States)

    Dores, C B; Milovancev, M; Russell, D S

    2018-03-01

    Radial sections are widely used to estimate adequacy of excision in canine cutaneous mast cell tumours (MCTs); however, this sectioning technique estimates only a small fraction of total margin circumference. This study aimed to compare histologic margin status in grade II/low grade MCTs sectioned using both radial and tangential sectioning techniques. A total of 43 circumferential margins were evaluated from 21 different tumours. Margins were first sectioned radially, followed by tangential sections. Tissues were examined by routine histopathology. Tangential margin status differed in 10 of 43 (23.3%) margins compared with their initial status on radial section. Of 39 margins, 9 (23.1%) categorized as histologic tumour-free margin (HTFM) >0 mm were positive on tangential sectioning. Tangential sections detected a significantly higher proportion of positive margins relative to radial sections (exact 2-tailed P-value = .0215). The HTFM was significantly longer in negative tangential margins than positive tangential margins (mean 10.1 vs 3.2 mm; P = .0008). A receiver operating characteristic curve comparing HTFM and tangentially negative margins found an area under the curve of 0.83 (95% confidence interval: 0.71-0.96). Although correct classification peaked at the sixth cut-point of HTFM ≥1 mm, radial sections still incorrectly classified 50% of margins as lacking tumour cells. Radial sections had 100% specificity for predicting negative tangential margins at a cut-point of 10.9 mm. These data indicate that for low grade MCTs, HTFMs >0 mm should not be considered completely excised, particularly when HTFM is <10.9 mm. This will inform future studies that use HTFM and overall excisional status as dependent variables in multivariable prognostic models. © 2017 John Wiley & Sons Ltd.

  16. Differential staining of bacteria: gram stain.

    Science.gov (United States)

    Moyes, Rita B; Reynolds, Jackie; Breakwell, Donald P

    2009-11-01

    In 1884, Hans Christian Gram, a Danish doctor, developed a differential staining technique that is still the cornerstone of bacterial identification and taxonomic division. This multistep, sequential staining protocol separates bacteria into four groups based on cell morphology and cell wall structure: Gram-positive cocci, Gram-negative cocci, Gram-positive rods, and Gram-negative rods. The Gram stain is useful for assessing bacterial contamination of tissue culture samples or for examining the Gram stain status and morphological features of bacteria isolated from mixed or isolated bacterial cultures. (c) 2009 by John Wiley & Sons, Inc.

  17. Port-Wine Stains

    Science.gov (United States)

    ... Safe Videos for Educators Search English Español Port-Wine Stains KidsHealth / For Parents / Port-Wine Stains What's ... Manchas de vino de oporto What Are Port-Wine Stains? A port-wine stain is a type ...

  18. A comparative study to evaluate liquid dish washing soap as an alternative to xylene and alcohol in deparaffinization and hematoxylin and eosin staining.

    Science.gov (United States)

    Pandey, Pinki; Dixit, Alok; Tanwar, Aparna; Sharma, Anuradha; Mittal, Sanjeev

    2014-07-01

    Our study presents a new deparaffinizing and hematoxylin and eosin (H and E) staining method that involves the use of easily available, nontoxic and eco-friendly liquid diluted dish washing soap (DWS) by completely eliminating expensive and hazardous xylene and alcohol from deparaffinizing and rehydration prior to staining, staining and from dehydration prior to mounting. The aim was to evaluate and compare the quality of liquid DWS treated xylene and alcohol free (XAF) sections with that of the conventional H and E sections. A total of 100 paraffin embedded tissue blocks from different tissues were included. From each tissue block, one section was stained with conventional H and E (normal sections) and the other with XAF H and E (soapy sections) staining method. Slides were scored using five parameters: Nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. Z-test was used for statistical analysis. Soapy sections scored better for cytoplasmic (90%) and crisp staining (95%) with a statistically significant difference. Whereas for uniformity of staining, normal sections (88%) scored over soapy sections (72%) (Z = 2.82, P 0.05). Liquid DWS is a safe and efficient alternative to xylene and alcohol in deparaffinization and routine H and E staining procedure. We are documenting this project that can be used as a model for other histology laboratories.

  19. A combined histological and MRI brain atlas of the common marmoset monkey, Callithrix jacchus.

    Science.gov (United States)

    Newman, John D; Kenkel, William M; Aronoff, Emily C; Bock, Nicholas A; Zametkin, Molly R; Silva, Afonso C

    2009-12-11

    The common marmoset, Callithrix jacchus, is of growing importance for research in neuroscience and related fields. In the present work, we describe a combined histological and magnetic resonance imaging (MRI) atlas constructed from the brains of two adult female marmosets. Histological sections were processed from Nissl staining and digitized to produce an atlas in a large format that facilitates visualization of structures with significant detail. Naming of identifiable brain structures was performed utilizing current terminology. The histological sections and a simplified schematic atlas are available online at http://udn.nichd.nih.gov/brainatlas_home.html.

  20. Accelerated staining technique using kitchen microwave oven

    Directory of Open Access Journals (Sweden)

    Archana Mukunda

    2015-01-01

    Full Text Available Introduction: Histopathological diagnosis of specimens is greatly dependent on good sample preparation and staining. Both of these processes is governed by diffusion of fluids and dyes in and out of the tissue, which is the key to staining. Diffusion of fluids can be accelerated by the application of heat that reduces the time of staining from hours to the minute. We modified an inexpensive model of kitchen microwave oven for staining. This study is an attempt to compare the reliability of this modified technique against the tested technique of routine staining so as to establish the kitchen microwave oven as a valuable diagnostic tool. Materials and Methods: Sixty different tissue blocks were used to prepare 20 pairs of slides for 4 different stains namely hematoxylin and eosin, Van Gieson′s, 0.1% toluidine blue and periodic acid-Schiff. From each tissue block, two bits of tissues were mounted on two different slides. One slide was stained routinely, and the other stained inside a microwave. A pathologist evaluated the stained slides and the results so obtained were analyzed statistically. Results: Microwave staining considerably cut down the staining time from hours to seconds. Microwave staining showed no loss of cellular and nuclear details, uniform-staining characteristics and was of excellent quality. Interpretation and Conclusion: The cellular details, nuclear details and staining characteristics of microwave stained tissues were better than or equal to the routine stained tissue. The overall quality of microwave-stained sections was found to be better than the routine stained tissue in majority of cases.

  1. Surface staining of small intestinal biopsies

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier

    1977-01-01

    Small intestinal biopsies are most often by routine examined under a stereo-microscope, prior to embedding for histological examination. This is done in order to get a view of the appearance of the mucosal pattern, especially villus configuration. The distinctness of the surface pattern however......, is improved considerably if the biopsies are stained with Alcian Green and/or PAS before they are examined. In the present paper a detailed description is given of staining of small intestinal biopsies as whole mounts. The difference between the unstained and the stained biopsies is illustrated by a few...

  2. Using Color and Grayscale Images to Teach Histology to Color-Deficient Medical Students

    Science.gov (United States)

    Rubin, Lindsay R.; Lackey, Wendy L.; Kennedy, Frances A.; Stephenson, Robert B.

    2009-01-01

    Examination of histologic and histopathologic microscopic sections relies upon differential colors provided by staining techniques, such as hematoxylin and eosin, to delineate normal tissue components and to identify pathologic alterations in these components. Given the prevalence of color deficiency (commonly called "color blindness")…

  3. Histological study of smoke extract of Tobacco nicotiana on the heart ...

    African Journals Online (AJOL)

    The heart, liver, lungs, kidney, and testes were carefully excised, blotted dry, and fixed in formol saline for histological analysis using Hematoxylin and Eosin stain. Results: Using the light microscope, it was observed that the histoarchitectural profiles of the studied organs in the sections obtained from the control animals ...

  4. A useful single-solution polychrome stain for plant material...Brook Cyte-Chrome I.

    Science.gov (United States)

    Stanley L Krugman; Julia F. Littlefield

    1968-01-01

    Fresh and chemically fixed sectioned plant material can be quickly stained by applying a Brook Cyte Chrome I polychrome stain. Staining time averaged only about 10 minutes. And exact timing of staining and de-staining is not as critical as with most of the commonly used stains. The overall quality is comparable to that of the traditional stains.

  5. Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains

    Science.gov (United States)

    Chen, Youting; Chen, Jing; Chen, Hong; Hong, Zhipeng; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Yanling; Chen, Jianxin

    2014-09-01

    Hematoxylin and eosin (H&E) staining of tissue samples is the standard approach in histopathology for imaging and diagnosing cancer. Recent reports have shown that multiphoton microscopy (MPM) provides better sample interface with single-cell resolution, which enhances traditional H&E staining and offers a powerful diagnostic tool with potential applications in oncology. The purpose of this study was to further expand the versatility of MPM by establishing the optical parameters required for imaging unstained histological sections of pancreatic neoplasms, thereby providing an efficient and environmentally sustainable alternative to H&E staining while improving the accuracy of pancreatic cancer diagnoses. We found that the high-resolution MPM images clearly distinguish between the structure of normal pancreatic tissues compared with pancreatic neoplasms in unstained histological sections, and discernable differences in tissue architecture and cell morphology between normal versus tumorigenic cells led to enhanced optical diagnosis of cancerous tissue. Moreover, quantitative assessment of the cytomorphological features visualized from MPM images showed significant differences in the nuclear-cytoplasmic ratios of pancreatic neoplasms compared with normal pancreas, as well as further distinguished pancreatic malignant tumors from benign tumors. These results indicate that the MPM could potentially serve as an optical tool for the diagnosis of pancreatic neoplasms in unstained histological sections.

  6. Stain Deconvolution Using Statistical Analysis of Multi-Resolution Stain Colour Representation.

    Directory of Open Access Journals (Sweden)

    Najah Alsubaie

    Full Text Available Stain colour estimation is a prominent factor of the analysis pipeline in most of histology image processing algorithms. Providing a reliable and efficient stain colour deconvolution approach is fundamental for robust algorithm. In this paper, we propose a novel method for stain colour deconvolution of histology images. This approach statistically analyses the multi-resolutional representation of the image to separate the independent observations out of the correlated ones. We then estimate the stain mixing matrix using filtered uncorrelated data. We conducted an extensive set of experiments to compare the proposed method to the recent state of the art methods and demonstrate the robustness of this approach using three different datasets of scanned slides, prepared in different labs using different scanners.

  7. leaves extracts as counter stain in gram staining reaction 56

    African Journals Online (AJOL)

    DR. AMINU

    is a stain with color contrasting to the principal stain, making the stained ... technology today, the Gram's staining method remains ... was aimed at employing the use of Henna leaves extract as ... fragrant, white or rose flowers in clusters. It is.

  8. Thick tissue diffusion model with binding to optimize topical staining in fluorescence breast cancer margin imaging

    Science.gov (United States)

    Xu, Xiaochun; Kang, Soyoung; Navarro-Comes, Eric; Wang, Yu; Liu, Jonathan T. C.; Tichauer, Kenneth M.

    2018-03-01

    Intraoperative tumor/surgical margin assessment is required to achieve higher tumor resection rate in breast-conserving surgery. Though current histology provides incomparable accuracy in margin assessment, thin tissue sectioning and the limited field of view of microscopy makes histology too time-consuming for intraoperative applications. If thick tissue, wide-field imaging can provide an acceptable assessment of tumor cells at the surface of resected tissues, an intraoperative protocol can be developed to guide the surgery and provide immediate feedback for surgeons. Topical staining of margins with cancer-targeted molecular imaging agents has the potential to provide the sensitivity needed to see microscopic cancer on a wide-field image; however, diffusion and nonspecific retention of imaging agents in thick tissue can significantly diminish tumor contrast with conventional methods. Here, we present a mathematical model to accurately simulate nonspecific retention, binding, and diffusion of imaging agents in thick tissue topical staining to guide and optimize future thick tissue staining and imaging protocol. In order to verify the accuracy and applicability of the model, diffusion profiles of cancer targeted and untargeted (control) nanoparticles at different staining times in A431 tumor xenografts were acquired for model comparison and tuning. The initial findings suggest the existence of nonspecific retention in the tissue, especially at the tissue surface. The simulator can be used to compare the effect of nonspecific retention, receptor binding and diffusion under various conditions (tissue type, imaging agent) and provides optimal staining and imaging protocols for targeted and control imaging agent.

  9. In vitro delineation of human brain-stem anatomy using a small resonator: correlation with macroscopic and histological findings

    International Nuclear Information System (INIS)

    Maeurer, J.; Mitrovic, T.; Knollmann, F.D.; Luedtke, E.; Requardt

    1996-01-01

    Our purpose was to investigate the potential of an experimental animal coil using a commercial MRI unit to delineate the anatomical structure of the human brain stem. Three formaldehyde-fixed brain-stem specimens were examined by MRI and sectioned perpendicular to their longitudinal axis. The images were compared with gross anatomy and myelin-stained histological sections. Fibre tracts and nuclei which were not evident on examination of the unstained specimen were readily identified by MRI. Due to its inherent grey/white matter contrast, MRI with a high-resolution coil delineates anatomical structures in a way comparable to the myelin-stained histological sections. However, pigmented structures, readily visible on examination of the unstained specimen were discernible on neither MRI nor on myelin-stained sections. The excellent anatomical detail and grey/white matter contrast provided by these images could make MRI a useful adjunct to the pathologist investigating brain disease. (orig.)

  10. Iron Stain on Wood

    Science.gov (United States)

    Mark Knaebe

    2013-01-01

    Iron stain, an unsightly blue–black or gray discoloration, can occur on nearly all woods. Oak, redwood, cypress, and cedar are particularly prone to iron stain because these woods contain large amounts of tannin-like extractives. The discoloration is caused by a chemical reaction between extractives in the wood and iron in steel products, such as nails, screws, and...

  11. Rapid in vivo vertical tissue sectioning by multiphoton tomography

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; König, Karsten

    2018-02-01

    A conventional tool in the pathological field is histology which involves the analysis of thin sections of tissue in which specific cellular structures are stained with different dyes. The process to obtain these stained tissue sections is time consuming and invasive as it requires tissue removal, fixation, sectioning, and staining. Moreover, imaging of live tissue is not possible. We demonstrate that multiphoton tomography can provide within seconds, non-invasive, label-free, vertical images of live tissue which are in quality similar to conventional light micrographs of histologic stained specimen. In contrast to conventional setups based on laser scanning which image horizontally sections, the vertical in vivo images are directly recorded by combined line scanning and timed adjustments of the height of the focusing optics. In addition, multiphoton tomography provides autofluorescence lifetimes which can be used to determine the metabolic states of cells.

  12. Modified paraffin wax for improvement of histological analysis efficiency.

    Science.gov (United States)

    Lim, Jin Ik; Lim, Kook-Jin; Choi, Jin-Young; Lee, Yong-Keun

    2010-08-01

    Paraffin wax is usually used as an embedding medium for histological analysis of natural tissue. However, it is not easy to obtain enough numbers of satisfactory sectioned slices because of the difference in mechanical properties between the paraffin and embedded tissue. We describe a modified paraffin wax that can improve the histological analysis efficiency of natural tissue, composed of paraffin and ethylene vinyl acetate (EVA) resin (0, 3, 5, and 10 wt %). Softening temperature of the paraffin/EVA media was similar to that of paraffin (50-60 degrees C). The paraffin/EVA media dissolved completely in xylene after 30 min at 50 degrees C. Physical properties such as the amount of load under the same compressive displacement, elastic recovery, and crystal intensity increased with increased EVA content. EVA medium (5 wt %) was regarded as an optimal composition, based on the sectioning efficiency measured by the numbers of unimpaired sectioned slices, amount of load under the same compressive displacement, and elastic recovery test. Based on the staining test of sectioned slices embedded in a 5 wt % EVA medium by hematoxylin and eosin (H&E), Masson trichrome (MT), and other staining tests, it was concluded that the modified paraffin wax can improve the histological analysis efficiency with various natural tissues. (c) 2010 Wiley-Liss, Inc.

  13. The use of special stains in liver biopsy interpretation: Implications ...

    African Journals Online (AJOL)

    Materials and Methods: The formalin fixed paraffin embedded blocks of liver biopsies reported in two histopathology laboratories between 2008 and 2013 were retrieved. These were stained with H and E and the following standard special stains for liver tissue histology – Perl's Prussian blue, reticulin, Sirius red, Shikata ...

  14. Histological analysis of extracranial carotid artery aneurysms.

    Directory of Open Access Journals (Sweden)

    Janna C Welleweerd

    Full Text Available Extracranial carotid artery aneurysms (ECAA are rare but may be accompanied with significant morbidity. Previous studies mostly focused on diagnostic imaging and treatment. In contrast, the pathophysiological mechanisms and natural course of ECAA are largely unknown. Understanding the pathophysiological background may add to prediction of risk for adverse outcome and need for surgical exclusion. The aim of this study was to investigate the histopathological characteristics of ECAA in patients who underwent complete surgical ECAA resection.From March 2004 till June 2013, 13 patients were treated with open ECAA repair. During surgery the aneurysm sac was resected and processed for standardized histological analysis. Sections were stained with routine hematoxylin and eosin and special stains to detect elastin, collagen, different types of inflammatory cells, vascular smooth muscle cells and endothelial cells.Histopathological characterization revealed two distinct categories: dissection (abrupt interruption of the media; n = 3 and degeneration (general loss of elastin fibers in the media; n = 10. In the degenerative samples the elastin fibers in the media were fragmented and were partly absent. Inflammatory cells were observed in the vessel wall of the aneurysms.Histological analysis in this small sample size revealed dissection and degeneration as the two distinct underlying mechanisms in ECAA formation.

  15. Accurate Depth of Radiofrequency-Induced Lesions in Renal Sympathetic Denervation Based on a Fine Histological Sectioning Approach in a Porcine Model.

    Science.gov (United States)

    Sakaoka, Atsushi; Terao, Hisako; Nakamura, Shintaro; Hagiwara, Hitomi; Furukawa, Toshihito; Matsumura, Kiyoshi; Sakakura, Kenichi

    2018-02-01

    Ablation lesion depth caused by radiofrequency-based renal denervation (RDN) was limited to radiofrequency-RDN cannot ablate a substantial percentage of renal sympathetic nerves. We aimed to define the true lesion depth achieved with radiofrequency-RDN using a fine sectioning method and to investigate biophysical parameters that could predict lesion depth. Radiofrequency was delivered to 87 sites in 14 renal arteries from 9 farm pigs at various ablation settings: 2, 4, 6, and 9 W for 60 seconds and 6 W for 120 seconds. Electric impedance and electrode temperature were recorded during ablation. At 7 days, 2470 histological sections were obtained from the treated arteries. Maximum lesion depth increased at 2 to 6 W, peaking at 6.53 (95% confidence interval, 4.27-8.78) mm under the 6 W/60 s condition. It was not augmented by greater power (9 W) or longer duration (120 seconds). There were statistically significant tendencies at 6 and 9 W, with higher injury scores in the media, nerves, arterioles, and fat. Maximum lesion depth was positively correlated with impedance reduction and peak electrode temperature (Pearson correlation coefficients were 0.59 and 0.53, respectively). Lesion depth was 6.5 mm for radiofrequency-RDN at 6 W/60 s. The impedance reduction and peak electrode temperature during ablation were closely associated with lesion depth. Hence, these biophysical parameters could provide prompt feedback during radiofrequency-RDN procedures in the clinical setting. © 2018 The Authors.

  16. Apoptosis detection in histological sections

    Czech Academy of Sciences Publication Activity Database

    Matalová, Eva; Dubská, Lenka; Míšek, Ivan

    2003-01-01

    Roč. 72, č. 7 (2003), s. 18-19 ISSN 0001-7213. [Congress of the European Association of Veterinary Anatomists/24./. 21.07.2002-25.07.2002, Brno] R&D Projects: GA ČR GP204/02/P112 Institutional research plan: CEZ:AV0Z5045916 Keywords : apoptosis Subject RIV: FF - HEENT, Dentistry

  17. Characterization of essential oil distribution in the root cross-section of Valeriana officinalis L. s.l. by using histological imaging techniques.

    Science.gov (United States)

    Penzkofer, Michael; Baron, Andrea; Naumann, Annette; Krähmer, Andrea; Schulz, Hartwig; Heuberger, Heidi

    2018-01-01

    The essential oil is an important compound of the root and rhizome of medicinally used valerian ( Valeriana officinalis L. s.l.), with a stated minimum content in the European pharmacopoeia. The essential oil is located in droplets, of which the position and distribution in the total root cross-section of different valerian varieties, root thicknesses and root horizons are determined in this study using an adapted fluorescence-microscopy and automatic imaging analysis method. The study was initiated by the following facts:A probable negative correlation between essential oil content and root thickness in selected single plants (elites), observed during the breeding of coarsely rooted valerian with high oil content.Higher essential oil content after careful hand-harvest and processing of the roots. In preliminary tests, the existence of oil containing droplets in the outer and inner regions of the valerian roots was confirmed by histological techniques and light-microscopy, as well as Fourier-transform infrared spectroscopy. Based on this, fluorescence-microscopy followed by image analysis of entire root cross-sections, showed that a large number of oil droplets (on average 43% of total oil droplets) are located close to the root surface. The remaining oil droplets are located in the inner regions (parenchyma) and showed varying density gradients from the inner to the outer regions depending on genotype, root thickness and harvesting depth. Fluorescence-microscopy is suitable to evaluate prevalence and distribution of essential oil droplets of valerian in entire root cross-sections. The oil droplet density gradient varies among genotypes. Genotypes with a linear rather than an exponential increase of oil droplet density from the inner to the outer parenchyma can be chosen for better stability during post-harvest processing. The negative correlation of essential oil content and root thickness as observed in our breeding material can be counteracted through a

  18. Detection of radioactively labeled proteins is quenched by silver staining methods: quenching is minimal for 14C and partially reversible for 3H with a photochemical stain

    International Nuclear Information System (INIS)

    Van Keuren, M.L.; Goldman, D.; Merril, C.R.

    1981-01-01

    Silver staining methods for protein detection in polyacrylamide gels have a quenching effect on autoradiography and fluorography. This effect was quantitated for proteins in two-dimensional gels by microdensitometry using a computer equipped with an image processor and by scintillation counting of proteins solubilized from the gels. The original histologically derived silver stain had a quenching effect that was severe and irreversible for 3 H detection and moderate for 14 C detection. A silver stain based on photochemical methods had minimal quenching of 14 C detection and less of a quenching effect than the histological stain for 3 H detection. The 3 H quenching effect was partially reversible for the photochemical stain

  19. Immunohistochemical expression of p53 proteins in Wilms' tumour: a possible association with the histological prognostic parameter of anaplasia.

    Science.gov (United States)

    Cheah, P L; Looi, L M; Chan, L L

    1996-01-01

    Wilms' tumour (nephroblastoma) has been associated with chromosomal abnormalities at the 11p13, 11p15 and 16q regions. A study into the possibility of mutations occurring within p53, the ubiquitous adult tumour suppressor gene, in Wilms' tumour was carried out. Thirty-eight cases were studied. Of these 36 were categorised into the favourable histology group and two into the unfavourable histology group based on the National Wilms' Tumour Study criteria. Archival formalin-fixed, paraffin-embedded tissue sections from each case were stained with a polyclonal (AB565:Chemicon) and a monoclonal (DO7:Dako) antibody raised against p53 protein using a peroxidase-labelled streptavidin biotin kit (Dako). 'Cure' (disease-free survival of 60 months or longer) was documented in 39% of cases with favourable histology tumours. Eleven percent in this group succumbed to the disease. Both cases with unfavourable histology died. Four out of 36 (11%) tumours with favourable histology demonstrated weak to moderate staining with both AB565 and DO7 in more than 75% of tumour cells. In contrast, p53 protein expression in unfavourable histology tumours was significantly increased compared with the favourable histology group (P = 0.021) with both cases demonstrating immunopositivity in > 75% of tumour cells when stained with AB565 and DO7. The intensity of staining ranged from moderate to strong in both cases. It appears from this preliminary study that the immunohistochemical expression of p53 protein in Wilms' tumour, presumably a result of mutation in the p53 tumour suppressor gene, correlates with histological classification, histological categorisation being one of the useful features in the prognostic assessment of Wilms' tumours.

  20. An optimized staining technique for the detection of Gram positive and Gram negative bacteria within tissue.

    Science.gov (United States)

    Becerra, Sandra C; Roy, Daniel C; Sanchez, Carlos J; Christy, Robert J; Burmeister, David M

    2016-04-12

    Bacterial infections are a common clinical problem in both acute and chronic wounds. With growing concerns over antibiotic resistance, treatment of bacterial infections should only occur after positive diagnosis. Currently, diagnosis is delayed due to lengthy culturing methods which may also fail to identify the presence of bacteria. While newer costly bacterial identification methods are being explored, a simple and inexpensive diagnostic tool would aid in immediate and accurate treatments for bacterial infections. Histologically, hematoxylin and eosin (H&E) and Gram stains have been employed, but are far from optimal when analyzing tissue samples due to non-specific staining. The goal of the current study was to develop a modification of the Gram stain that enhances the contrast between bacteria and host tissue. A modified Gram stain was developed and tested as an alternative to Gram stain that improves the contrast between Gram positive bacteria, Gram negative bacteria and host tissue. Initially, clinically relevant strains of Pseudomonas aeruginosa and Staphylococcus aureus were visualized in vitro and in biopsies of infected, porcine burns using routine Gram stain, and immunohistochemistry techniques involving bacterial strain-specific fluorescent antibodies as validation tools. H&E and Gram stain of serial biopsy sections were then compared to a modification of the Gram stain incorporating a counterstain that highlights collagen found in tissue. The modified Gram stain clearly identified both Gram positive and Gram negative bacteria, and when compared to H&E or Gram stain alone provided excellent contrast between bacteria and non-viable burn eschar. Moreover, when applied to surgical biopsies from patients that underwent burn debridement this technique was able to clearly detect bacterial morphology within host tissue. We describe a modification of the Gram stain that provides improved contrast of Gram positive and Gram negative microorganisms within host

  1. A novel non-toxic xylene substitute (SBO) for histology.

    Science.gov (United States)

    Kunhua, Wang; Chuming, Fan; Tao, Lai; Yanmei, Yang; Xin, Yang; Xiaoming, Zhang; Xuezhong, Guo; Xun, Lai

    2012-01-01

    Xylene has been generally used as a clearing and deparaffinizing agent in histology. Because of the potential toxic and flammable nature of xylene, its substitutes have been introduced into some laboratories. In this study, we introduced a novel, non-toxic xylene substitute (SBO), which was generated through a mixture of 86% of white oil No.2 and 14% of N-heptane. SBO had a high boiling point (188°C) and flash point (144°C) coupled with a scentless and decreased volatility. To compare the effectiveness of SBO and xylene in histology, a wide range of tissue samples from rats and human beings were processed in parallel in SBO and xylene, subjected to various staining procedures. Similar to the xylene-processed paraffin blocks, the SBO-processed counterparts were easy to section without any evidence of cell shrinkage. Assessment of the SBO-treated sections stained with hematoxylin-eosin revealed a good maintenance of cell morphology and structure, and a clear definition of the cytoplasm and the nucleus. Moreover, comparable good results were achieved between the SBO- and xylene-processed tissues in other histochemical and immunohistochemical stainings. Six-month clinical applications at one department of pathology supported the potentials of SBO as a xylene substitute. In conclusion, we suggest that SBO is a safe and efficient substitute of xylene and may probably replace xylene without losing valuable diagnostic information.

  2. Port-wine stain

    Science.gov (United States)

    ... About MedlinePlus Show Search Search MedlinePlus GO GO About MedlinePlus Site Map FAQs Customer Support Health Topics Drugs & Supplements Videos & Tools Español You Are Here: Home → Medical Encyclopedia → Port-wine stain URL of this page: //medlineplus.gov/ency/ ...

  3. Stool Gram stain

    Science.gov (United States)

    ... stool sample. The Gram stain method is sometimes used to quickly diagnose bacterial infections. How the Test is Performed You will need to collect a stool sample. There are many ways to collect the sample. You can catch the stool on plastic wrap that is loosely placed over the toilet bowl ...

  4. Stained Glass and Flu

    Centers for Disease Control (CDC) Podcasts

    2017-02-01

    Dr. Robert Webster, an Emeritus member of the Department of Infectious Diseases at St. Jude Children's Research Hospital, discusses his cover art story on stained glass and influenza.  Created: 2/1/2017 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 2/1/2017.

  5. Comparison of special stains for keratin with routine hematoxylin and eosin stain.

    Science.gov (United States)

    Rao, Roopa S; Patil, Shankargouda; Majumdar, Barnali; Oswal, Rakesh G

    2015-03-01

    Keratins are the most abundant proteins and are characteristic findings in many epithelial pathologies, making it a diagnostically important marker, both histopathologically and immunohistochemically. Since, immunohistochemistry is an expensive diagnostic tool, special stains to detect the degree of keratinization could serve as a faster and economic option. The aim of the present study was to compare the efficacy of special stains for keratin with standard hematoxylin and eosin stain (H and E). Objectives include: (i) To subject the diagnosed cases of keratin disorders to the selected special stains: Ayoub-shklar method, Dane-Herman method, Alcian blue -periodic acid Schiff 's (PAS), rapid papanicolaou (PAP) and Gram's stain. (ii) To compare the staining specificity and staining intensity of special stains with respect to routine hematoxylin and eosin (H and E) stain. (iii) To compare the efficacy of special stains to routine H and E stain in identification of the type of keratin present in the selected cases. A total of 80 cases of known pathology for keratin were retrieved from the department archive, which included 10 each of normal gingiva, hyperkeratosis, squamous papilloma, verrucous hyperplasia, verrucous carcinoma, well-differentiated squamous cell carcinoma, orthokeratinized odontogenic cyst and keratocystic odontogenic tumors. Six sections of 4 µ each from the paraffin blocks were made, stained with H and E and the special stains and these were evaluated by 2 pathologists based on the modified scoring criteria from Rahma Al-Maaini and Philip Bryant 2008. The results were tabulated using Chi square and kappa statistics. The statistical values for identification of the type of keratinization was insignificant showing that ortho and parakeratinized epithelia could be correctly identified by both H and E as well as all the special stains. Furthermore, all the special stains showed a positive result and statistical significance (P < 0.001) with respect to

  6. Technique of neutron-induced (fission-track) autoradiography with histological detail

    International Nuclear Information System (INIS)

    Smith, J.M.; Taylor, G.N.; Jee, W.S.S.

    1980-01-01

    The primary advantage of neutron-induced or fission-track autoradiography compared with conventional autoradiography is that for low concentrations of fissile nuclides prohibitively long exposure times may be avoided. However, it is difficult to produce imaging of biological structures on the neutron-induced autoradiograph which would allow localization of the nuclide histologically. The technique presented circumvents this difficulty using a thin polycarbonate film applied to the histologically stained tissue section mounted on a quartz substrate. After irradiation of the tissue section with an appropriate thermal neutron flux, the fission fragment tracks are revealed by etching the film with KOH. The tracks, superimposed on the stained tissue, may be observed under the light microscope in the same manner as for conventional nuclear emulsion autoradiography

  7. 7 CFR 28.442 - Middling Yellow Stained Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] below color grade cotton ...

  8. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] ...

  9. The histological lesions of Trichophyton mentagrophytes var erinacei infection in dogs.

    Science.gov (United States)

    Fairley, R A

    2001-04-01

    A retrospective study of the histological features of four cases of canine Trichophyton mentagrophytes var erinacei infection is reported. In all four dogs the initial lesions affected the dorsal muzzle and in two dogs the lesions spread to more distant sites on the body. Clinically, the lesions were characterized by scaling, crusting and hair loss. Histologically, the main lesions were characterized by acanthosis, epidermal, ostial and infundibular hyperkeratosis, serocellular crusting, mural folliculitis and furunculosis. Fungal hyphae were usually sparse and often difficult to see in haematoxylin and eosin stained sections. When visible they were seen in the epidermal, ostial and infundibular scale and, less frequently, within hair shafts.

  10. Modified Field's staining--a rapid stain for Trichomonas vaginalis.

    Science.gov (United States)

    Afzan, M Yusuf; Sivanandam, S; Kumar, G Suresh

    2010-10-01

    Trichomonas vaginalis, a flagellate protozoan parasite commonly found in the human genitourinary tract, is transmitted primarily by sexual intercourse. Diagnosis is usually by in vitro culture method and staining with Giemsa stain. There are laboratories that use Gram stain as well. We compared the use of modified Field's (MF), Giemsa, and Gram stains on 2 axenic and xenic isolates of T. vaginalis, respectively. Three smears from every sediment of spun cultures of all 4 isolates were stained, respectively, with each of the stains. We showed that MF staining, apart from being a rapid stain (20 s), confers sharper staining contrast, which differentiates the nucleus and the cytoplasm of the organism when compared to Giemsa and Gram staining especially on parasites from spiked urine samples. The alternative staining procedure offers in a diagnostic setting a rapid stain that can easily visualize the parasite with sharp contrasting characteristics between organelles especially the nucleus and cytoplasm. Vacuoles are more clearly visible in parasites stained with MF than when stained with Giemsa. Copyright © 2010 Elsevier Inc. All rights reserved.

  11. Modified Genta triple stain for identifying Helicobacter pylori.

    OpenAIRE

    el-Zimaity, H M; Wu, J; Graham, D Y

    1999-01-01

    AIM: To evaluate whether lead nitrate could replace uranyl nitrate in the Genta stain for H pylori without sacrificing the advantages of the triple stain (Steiner silver impregnation combined with Alcian blue and haematoxylin/eosin (H&E)). METHODS: A comparison was made in 16 specimens between the original triple stain and the revised version. One pathologist evaluated all sections. RESULTS: Direct substitution of lead nitrate for uranium nitrate produced well stained organisms without interf...

  12. Stented Vessels: A Challenge for Histological Preparation and Microscopy

    Directory of Open Access Journals (Sweden)

    Andrea Nolte

    2013-06-01

    Full Text Available Objective: The first procedure to treat blocked coronary arteries was coronary artery bypass graft surgery. In 1977, Andreas Gruntzig introduced percutaneous transluminal coronary angioplasty (PTCA. Today, several stent systems exist ranging from bare metal stents to various drug-eluting stents. Unfortunately, our understanding of the arterial reaction to stent implantation is incomplete – primarily due to technical limitations in the histological study of stented vascular tissue. Methods: In our study, we examined different histological preparation methods based on the embedding material methacrylate. The procedure of embedding and sectioning stented porcine arteries was optimized for the specific requirements, like histochemistry, immunohistochemistry or pre-stained fluorescence. Furthermore, we used a microscopical technique described as fluorescence intensity decay shape analysis microscopy (FIDSAM to eliminate auto-fluorescence from fluorescently labeled tissue. Results: The sections were suitable for histochemical and immunohistochemical staining. Additionally, pre-labeled fluorescence in the porcine tissue was not lost by the embedding process. The evaluation of arterial cross sections with FIDSAM technology gave new, very important insights into the examination possibilities of fluorescently labeled tissue. Conclusions: Future studies of the vascular response to a variety of new stent materials will provide important clues to the pathogenesis resulting in restenosis and occlusion of stents. [J Interdiscipl Histopathol 2013; 1(3.000: 104-112

  13. New Grocott Stain without Using Chromic Acid

    International Nuclear Information System (INIS)

    Shiogama, Kazuya; Kitazawa, Kayo; Mizutani, Yasuyoshi; Onouchi, Takanori; Inada, Ken-ichi; Tsutsumi, Yutaka

    2015-01-01

    We established a new “ecological” Grocott stain for demonstrating fungi, based upon a 4R principle of refusal, reduction, reuse, and recycle of waste management. Conventional Grocott stain employs environmentally harsh 5% chromic acid for oxidization. Initially, we succeeded in reducing the concentration of chromic acid from 5% to 1% by incubating the solution at 60°C and using five-fold diluted chromic acid solution at which point it was reusable. Eventually, we reached the refusal level where 1% periodic acid oxidization was efficient enough, when combined with preheating of sections in the electric jar, microwave oven, or pressure pan. For convenience sake, we recommend pressure pan heating in tap water for 10 min. Stainability of fungi in candidiasis and aspergillosis was comparable with conventional Grocott stain, while Mucor hyphae showed enhanced staining. The modified sequence was further applicable to detecting a variety of mycotic pathogens in paraffin sections. Our environmentally-friendly Grocott stain also has the advantage of avoiding risk of human exposure to hexavalent chromium solution in the histopathology laboratory. The simple stain sequence is can be easily applied worldwide

  14. Enhanced-Depth Imaging Optical Coherence Tomography of the Human Choroid In Vivo Compared With Histology After Enucleation

    DEFF Research Database (Denmark)

    Li, Xiao Qiang; Heegaard, Steffen; Kiilgaard, Jens Folke

    2016-01-01

    PURPOSE: This study compared in vivo enhanced-depth imaging optical coherence tomography (EDI-OCT) with ex vivo histology of the choroid in human eyes. METHODS: Three eyes in three patients with advanced iris melanoma without posterior segment involvement underwent EDI-OCT less than 24 hours prior...... to enucleation and, in one eye, immediately after enucleation. Following fixation in 4% buffered formaldehyde and paraffin embedding, serial sections of the whole eye were cut horizontally, mounted, stained with hematoxylin-eosin and digitized. Alignment between histology and EDI-OCT was made on landmarks...... and subfoveal choroid thickness reduced to 56%, 45%, and 56%, respectively, of its in vivo thickness on EDI-OCT. CONCLUSIONS: There were no identifiable discrepancies in choroidal structural patterns between clinical EDI-OCT and histologic sections except that after enucleation and histologic fixation choroidal...

  15. HISTOLOGICAL CHARACTERISTICS OF THE QUILTY EFFECT

    Directory of Open Access Journals (Sweden)

    I. M. Iljinsky

    2015-01-01

    Full Text Available Aim. Identifi cation of possible histological differences of the Quilty effect in acute rejection and its absence as well as studying the proliferation of blood and lymph vessels in the area of Quilty damage.Materials and methods. 883 endomyocardial biopsy materials from 352 patients were studied. Histological sections were stained with hematoxylin and eosin, Masson method; the endothelium of lymphatic vessels was stained with immunoperoxidase method using a marker D2-40.Results. The Quilty effect was observed both in acute rejection and inits absence. In the majority of cases the Quilty effect was of type «A» and it was combined with acute rejection. The Quilty effect of type «B» has been mostly in the G2R. Acute rejection is characterized by diffuse form of endocardium lymphoid infi ltration. Follicular form resembles lymphoid organ tissue. Different types and forms of the Quilty effect may be combined in the same biopsy. Proliferation of blood vessels presents in the area of the Quilty effect. Lymphatic vessels are missing in endocardium and in the area of the Quilty effect. They could be found only in the myocardium of endomiocardial biopsy.Conclusion. Diffuse lymphoid infi ltration of the endocardium is a characteristic feature of acute rejection of the transplanted heart. Follicular form of the Quilty effect is similar to lymphoid tissue whose role requires further study using immunohistochemical methods.

  16. Automated recognition of cell phenotypes in histology images based on membrane- and nuclei-targeting biomarkers

    International Nuclear Information System (INIS)

    Karaçalı, Bilge; Vamvakidou, Alexandra P; Tözeren, Aydın

    2007-01-01

    Three-dimensional in vitro culture of cancer cells are used to predict the effects of prospective anti-cancer drugs in vivo. In this study, we present an automated image analysis protocol for detailed morphological protein marker profiling of tumoroid cross section images. Histologic cross sections of breast tumoroids developed in co-culture suspensions of breast cancer cell lines, stained for E-cadherin and progesterone receptor, were digitized and pixels in these images were classified into five categories using k-means clustering. Automated segmentation was used to identify image regions composed of cells expressing a given biomarker. Synthesized images were created to check the accuracy of the image processing system. Accuracy of automated segmentation was over 95% in identifying regions of interest in synthesized images. Image analysis of adjacent histology slides stained, respectively, for Ecad and PR, accurately predicted regions of different cell phenotypes. Image analysis of tumoroid cross sections from different tumoroids obtained under the same co-culture conditions indicated the variation of cellular composition from one tumoroid to another. Variations in the compositions of cross sections obtained from the same tumoroid were established by parallel analysis of Ecad and PR-stained cross section images. Proposed image analysis methods offer standardized high throughput profiling of molecular anatomy of tumoroids based on both membrane and nuclei markers that is suitable to rapid large scale investigations of anti-cancer compounds for drug development

  17. Hidradenocarcinoma presenting as soft tissue mass: Case report with cytomorphologic description, histologic correlation, and differential diagnosis.

    Science.gov (United States)

    Jinnah, Alexander H; Emory, Cynthia L; Mai, Nicholas H; Bergman, Simon; Salih, Ziyan T

    2016-05-01

    Hidradenocarcinoma (HAC) is a rare adenexal tumor with a propensity for the head and neck region and extremities. We report a case of hidradenocarcinnoma in a 56-year-old woman with a mass on her right palm sampled by fine-needle aspiration and later confirmed on histological examination. Fine-needle aspiration cytology revealed a dual population of cells including polyhedral eosinophilic cells and glycogen containing cells with pale/clear cytoplasm. The nuclei were pleomorphic with prominent nucleoli. Occassional papillary structures were identified on the cell block material. A series of immunohistochemical stains were performed and an adnexal neoplasm was suggested. The mass was resected. On histologic sections, infiltration into the adjacent soft tissue was identified. After an additional series of immunohistochemical stains, the diagnosis was confirmed as a HAC. Herein, we present our findings and discuss the differential diagnoses. © 2016 Wiley Periodicals, Inc.

  18. Histology of a Woolly Mammoth (Mammuthus primigenius) Preserved in Permafrost, Yamal Peninsula, Northwest Siberia.

    Science.gov (United States)

    Papageorgopoulou, Christina; Link, Karl; Rühli, Frank J

    2015-06-01

    In 2007, the baby woolly mammoth (Mammuthus primigenius) named Lyuba was found frozen in the Siberian tundra permafrost along the Yuribey River. She was proclaimed the best-preserved mammoth discovery. As part of the endoscopic examination of Lyuba, tissue samples of hair, muscle, and internal organs were taken. The sectioned biopsies were stained using standard and special histological stains. In general, the microscopic preservation of the tissue was good although no clearly identifiable cell nuclei were found by standard staining methods. Only a few cell nuclei could be identified in some samples when fluorescence stained with DAPI. The best-preserved structures were collagen fibers and muscle tissue, which gave some structural resemblance to the organs. In the hairs, evidence of pigmentation, a scaly surface, diagonal intra-hair structures, and a medulla were seen. Fat droplets could be identified with Sudan Red in the subcutaneous fat sample and in several organs. Bacteria were seen on the lumen side of the small intestine and caecum, and in the liver and lung tissue. In addition, fungi and pollen were seen in the lung sample. In the wall of the caecum and small intestine, blood vessels and nerves were visualized. Iron was identified in the vivianite sample. Some biopsies compared well structurally with the African elephant tissue sections. The histological findings support the theory that Lyuba drowned in muddy water. The microscopic tissue preservation and cell nuclei destruction indicate that Lyuba's body underwent at least one freeze-thaw cycle. © 2015 Wiley Periodicals, Inc.

  19. Stain-free histopathology by programmable supercontinuum pulses

    DEFF Research Database (Denmark)

    Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry

    2016-01-01

    The preparation, staining, visualization and interpretation of histological images of tissue is well accepted as the gold standard process for the diagnosis of disease. These methods have a long history of development, and are used ubiquitously in pathology, despite being highly time- and labour-...

  20. The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections.

    Science.gov (United States)

    Kobayashi, Naomi; Bauer, Thomas W; Tuohy, Marion J; Lieberman, Isador H; Krebs, Viktor; Togawa, Daisuke; Fujishiro, Takaaki; Procop, Gary W

    2006-08-01

    We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of "molecular Gram stain" with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results.

  1. Histopathological evaluation of ocular microsporidiosis by different stains

    Directory of Open Access Journals (Sweden)

    Sharma Savitri

    2006-06-01

    Full Text Available Abstract Background There is limited data on comparing stains in the detection of microsporidia in corneal biopsies. Hence we wanted to evaluate various stains for their ability to detect microsporidia in corneal tissue sections. Methods Four cases diagnosed with microsporidiosis on Hematoxylin and Eosin and Periodic Acid Schiff's stained sections of the corneal button between January 2002 and December 2004, were included. Further sections were prospectively stained with calcofluor white, Gram, Giemsa, Masson's trichrome, acridine orange, Gomori's methenamine silver, Gram's chromotrope and modified acid fast stain. The stained sections were analyzed for the spore characteristics in terms of size, shape, color contrast, cell wall morphology, waist band in cytoplasm and ease of detection. Results All sections showed microsporidial spores as 3 – 5 μm, oval bodies. 1% acid fast, Gram's chromotrope and GMS stains provided a reliable diagnosis of microsporidia as diagnostic waist band could be identified and good contrast helped distinguish the spores from inflammatory debris. Conclusion Considering the ease of performance, cost effectiveness and rapidity of the technique, 1% acid fast stain and Gram's chromotrope stain are ideal for the detection of microsporidia.

  2. Investigating the influence of standard staining procedures on the copper distribution and concentration in Wilson's disease liver samples by laser ablation-inductively coupled plasma-mass spectrometry.

    Science.gov (United States)

    Hachmöller, Oliver; Aichler, Michaela; Schwamborn, Kristina; Lutz, Lisa; Werner, Martin; Sperling, Michael; Walch, Axel; Karst, Uwe

    2017-12-01

    The influence of rhodanine and haematoxylin and eosin (HE) staining on the copper distribution and concentration in liver needle biopsy samples originating from patients with Wilson's disease (WD), a rare autosomal recessive inherited disorder of the copper metabolism, is investigated. In contemporary diagnostic of WD, rhodanine staining is used for histopathology, since rhodanine and copper are forming a red to orange-red complex, which can be recognized in the liver tissue using a microscope. In this paper, a laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) method is applied for the analysis of eight different WD liver samples. Apart from a spatially resolved elemental detection as qualitative information, this LA-ICP-MS method offers also quantitative information by external calibration with matrix-matched gelatine standards. The sample set of this work included an unstained and a rhodanine stained section of each WD liver sample. While unstained sections of WD liver samples showed very distinct structures of the copper distribution with high copper concentrations, rhodanine stained sections revealed a blurred copper distribution with significant decreased concentrations in a range from 20 to more than 90%. This implies a copper removal from the liver tissue by complexation during the rhodanine staining. In contrast to this, a further HE stained sample of one WD liver sample did not show a significant decrease in the copper concentration and influence on the copper distribution in comparison to the unstained section. Therefore, HE staining can be combined with the analysis by means of LA-ICP-MS in two successive steps from one thin section of a biopsy specimen. This allows further information to be gained on the elemental distribution by LA-ICP-MS additional to results obtained by histological staining. Copyright © 2017 Elsevier GmbH. All rights reserved.

  3. Anatomy and Histology of an Epicanthal Fold.

    Science.gov (United States)

    Park, Jae Woo; Hwang, Kun

    2016-06-01

    The aim of this study is to elucidate the precise anatomical and histological detail of the epicanthal fold.Thirty-two hemifaces of 16 Korean adult cadavers were used in this study (30 hemifaces with an epicanthal fold, 2 without an epicanthal fold). In 2 patients who had an epicanthoplasty, the epicanthal folds were sampled.In a dissection, the periorbital skin and subcutaneous tissues were removed and the epicanthal fold was observed in relation to each part of the orbicularis oculi muscle. Specimens including the epicanthal fold were embeddedin in paraffin, sectioned at 10 um, and stained with Hematoxylin-Eosin. The horizontal section in the level of the paplebral fissure was made and the prepared slides were observed under a light microscope.In the specimens without an epicanthal fold, no connection between the upper preseptal muscle and the lower preseptal muscle was found. In the specimens with an epicanthal fold, a connection of the upper preseptal muscle to the lower preseptal muscle was observed. It was present in all 15 hemifaces (100%). There was no connection between the pretarsal muscles. In a horizontal section, the epicanthal fold was composed of 3 compartments: an outer skin lining, a core structure, and an innerskin lining. The core structure was mainly composed of muscular fibers and fibrotic tissue and they were intermingled.Surgeons should be aware of the anatomical details of an epicanthal fold. In removing or reconstructing an epicanthal fold, the fibromuscular core band should also be removed or reconstructed.

  4. Touch cytology in diagnosing Helicobacter pylori: comparison of four staining methods.

    Science.gov (United States)

    Hashemi, M R; Rahnavardi, M; Bikdeli, B; Dehghani Zahedani, M; Iranmanesh, F

    2008-06-01

    Helicobacter pylori (Hp), a major cause of peptic ulcer disease and an important risk factor for gastric malignancy, can be diagnosed by several methods. Touch cytology (TC) of the gastric mucosa has been noted to give good results and has been found to be very simple, inexpensive and rapid. However, evidence regarding the accuracy of different staining methods of TC is lacking. The present study aims at defining the diagnostic accuracy of four different staining methods of TC. Biopsy specimens were taken from the antral mucosa of one hundred consecutive patients referred for upper gastrointestinal endoscopy (UGIE) for various indications. TC slides were processed by four staining methods: Wright, Giemsa, Papanicolaou and Gram. Rapid urease test (RUT) and histological examination of specimens were also performed. The same experienced pathologist evaluated the coded samples. A patient's Hp status was established by minimum concordance of the three tests, including histology, RUT, and 'Touch mean'. The latter was defined positive when at least three of the four TC staining methods were positive. Forty-six patients (46%) were positive for Hp according to Hp status. TC stained by Wright had excellent agreement with both histology (kappa = 0.80, P stained TC (88.89%) was significantly more specific than both Giemsa- (74.07%; P stained (70.37%; P stained TC can safely substitute for histology. However, when assessment for severity of mucosal damage or cell atypias is meant, histology cannot be neglected.

  5. HISTOLOGIC, IMMUNOHISTOCHEMICAL, AND ELECTRON MICROSCOPIC CHARACTERIZATION OF A MALIGNANT IRIDOPHOROMA IN A DWARF BEARDED DRAGON (POGONA HENRYLAWSONI).

    Science.gov (United States)

    de Brot, Simone; Sydler, Titus; Nufer, Lisbeth; Ruetten, Maja

    2015-09-01

    A dwarf bearded dragon (Pogona henrylawsoni) was presented with a white subcutaneous mandibular mass and multiple nodules in the oral mucosa, heart, liver, kidney, intestine, and visceral fat. Histologically, the tumor consisted of densely packed spindle-shaped cells with brow intracytoplasmic pigment that exhibited white-blue birefringence with polarized light. Immunohistochemical staining was negative for S-100 and weakly positive with melan A. Electron microscopic examination revealed cytoplasmic irregular and oblong empty spaces, laminated and often arranged into short stacks, compatible with reflecting platelet profiles typically seen in iridophores. However, in unstained ultrathin sections, electron-dense crystalline material was present, which filled the empty spaces described for stained sections before. Based on histology, immunohistochemistry, and biologic behavior, a malignant iridophoroma was diagnosed. To the authors' knowledge, iridophoromas in lizards have rarely been characterized by using electronic microscopy. Moreover, this is the first description of an iridophoroma in a dwarf bearded dragon.

  6. Improved Nissl method to stain formaldehyde or glutaraldehyde-fixed material.

    Science.gov (United States)

    Böck, P

    1979-05-15

    Nissl staining of paraffin sections from formaldehyde- or glutaraldehyde-fixed specimens is significantly intensified when sections are kept in a 50% (w/v) aqueous solution of potassium metabisulfite before being stained by a conventional Nissl method.

  7. eHistology image and annotation data from the Kaufman Atlas of Mouse Development.

    Science.gov (United States)

    Baldock, Richard A; Armit, Chris

    2017-12-20

    "The Atlas of Mouse Development" by Kaufman is a classic paper atlas that is the de facto standard for the definition of mouse embryo anatomy in the context of standard histological images. We have re-digitised the original H&E stained tissue sections used for the book at high resolution and transferred the hand-drawn annotations to digital form. We have augmented the annotations with standard ontological assignments (EMAPA anatomy) and made the data freely available via an online viewer (eHistology) and from the University of Edinburgh DataShare archive. The dataset captures and preserves the definitive anatomical knowledge of the original atlas, provides a core image set for deeper community annotation and teaching, and delivers a unique high-quality set of high-resolution histological images through mammalian development for manual and automated analysis. © The Authors 2017. Published by Oxford University Press.

  8. Smoking is associated with severity of liver fibrosis but not with histological severity in nonalcoholic fatty liver disease. Results from a cross-sectional study.

    Science.gov (United States)

    Munsterman, Isabelle D; Smits, Mark M; Andriessen, Rene; van Nieuwkerk, Carin M J; Bloemena, Elisabeth; Mulder, Chris J J; Tjwa, Eric T T L; van Geenen, Erwin J M

    2017-08-01

    To assess the influence of smoking on histological disease severity and fibrosis in real-world NAFLD patients. Consecutive NAFLD patients were identified with liver biopsies performed between 2008 and 2015. Characteristics such as smoking status and total number of pack years were collected. Biopsies were revised and BRUNT fibrosis and NAFLD activity score (NAS) determined. Patients with a high NAS (≥5) were compared to patients with a low NAS (smoking (current or past smoker) were defined ever smokers. Fifty-six patients were included (mean age 49 ± 14.3, 68.9% males and 39.3% history of smoking). Ever smokers had a higher fibrosis score than never smokers; two (IQR 0-3) versus one (IQR 1-1.5) (p = .040). Patients with advanced fibrosis smoked significantly more pack years than patients with no-early fibrosis; 10.6 (IQR 0-25.8) versus 0 (IQR 0-7) (p = .011). There is a weak to moderate correlation between fibrosis stage and number of pack years (Spearman's Rho = 0.341, p = .012). There was no difference in NAS between never and ever smokers; 2.8 ± 1.5 versus 3.3 ± 1.4 (p = .205). Patients with NAS Smoking is associated with severity of NAFLD-related liver fibrosis but not with histological disease severity. This supports the recommendation to cease smoking for NAFLD patients.

  9. Hirschsprung's disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining

    Science.gov (United States)

    Memarzadeh, Mehrdad; Talebi, Ardeshir; Edalaty, Masod; Hosseinpour, Mehrdad; Vahidi, Nasrin

    2009-01-01

    Background: The diagnosis of Hirschsprung's disease (HD) is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E) as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods: In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens) were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staining slides were compared with immunohistochemical (IHC) staining including: S100, NSE, CD117, CD56, Cathepsin D, Vimentin, BCL2, GFAP, Synaptophysin and chromogranin. Results: The study revealed 30 negative (absence of ganglion cells) cases (55.5%), 17 positive cases (31.04%) and seven suspected cases (12.9%) of ganglion cells on the H and E staining. On IHC staining with CD56 and Cathepsin D, all of the 17 positive cases detected through H and E, were confirmed for having ganglion cells and out of 30 cases reported negative on H and E staining, 28(93.3%) were reported negative and two (6.7%) positive by IHC staining. Of the seven suspected cases H and E staining), IHC staining detectedganglion cells only in five slides; two remained negative. Conclusions: IHC staining using CD56 and Cathepsin D improved the accuracy of diagnosis in HD when used in addition to H and E staining technique, especially for negative or suspicious slides. PMID:20671847

  10. Melanin fate in the human epidermis: a reassessment of how best to detect and analyse histologically.

    Science.gov (United States)

    Joly-Tonetti, Nicolas; Wibawa, Judata I D; Bell, Mike; Tobin, Desmond

    2016-07-01

    Melanin is the predominant pigment responsible for skin colour and is synthesized by the melanocyte in the basal layer of the epidermis and then transferred to surrounding keratinocytes. Despite its optical properties, melanin is barely detectable in unstained sections of human epidermis. However, identification and localization of melanin is of importance for the study of skin pigmentation in health and disease. Current methods for the histologic quantification of melanin are suboptimal and are associated with significant risk of misinterpretation. The aim of this study was to reassess the existing literature and to develop a more effective histological method of melanin quantification in human skin. Moreover, we confirm that Warthin-Starry (WS) stain provides a much more sensitive and more specific melanin detection method than the commonplace Fontana-Masson (FM) stain. For example, WS staining sensitivity allowed the visualization of melanin even in very pale Caucasian skin that was missed by FM or Von Kossa (VK) stains. From our reassessment of the histology-related literature, we conclude that so-called melanin dust is most likely an artifact of discoloration due to non-specific silver deposition in the stratum corneum. Unlike FM and VK, WS was not associated with this non-specific stratum corneum darkening, misinterpreted previously as 'degraded' melanin. Finally, WS melanin particle counts were largely similar to previously reported manual counts by transmission electron microscopy, in contrast to both FM and VK. Together these findings allow us to propose a new histology/Image J-informed method for the accurate and precise quantification of epidermal melanin in skin. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Histological changes in the pancreas following administration of ...

    African Journals Online (AJOL)

    The experiment lasted for 28 days. The animals were anaesthetized using chloroform inhalation and the peritoneum stripped open and the pancreas removed and prepared for histological observation using haematoxylin and eosin staining technique. Histology showed regenerative changes of pancreatic islet cell at a dose ...

  12. Say goodbye to coffee stains

    NARCIS (Netherlands)

    Eral, Burak; van den Ende, Henricus T.M.; Mugele, Friedrich Gunther

    2012-01-01

    Discussing ideas over a mug of coffee or tea is the lifeblood of science, but have you ever thought about the stains that can be inadvertently left behind? H Burak Eral, Dirk van den Ende and Frieder Mugele explain how these stains, which can be a major annoyance in some biology techniques, can be

  13. Differentiation of osteophyte types in osteoarthritis - proposal of a histological classification.

    Science.gov (United States)

    Junker, Susann; Krumbholz, Grit; Frommer, Klaus W; Rehart, Stefan; Steinmeyer, Jürgen; Rickert, Markus; Schett, Georg; Müller-Ladner, Ulf; Neumann, Elena

    2016-01-01

    Osteoarthritis is not only characterized by cartilage degradation but also involves subchondral bone remodeling and osteophyte formation. Osteophytes are fibrocartilage-capped bony outgrowths originating from the periosteum. The pathophysiology of osteophyte formation is not completely understood. Yet, different research approaches are under way. Therefore, a histological osteophyte classification to achieve comparable results in osteophyte research was established for application to basic science research questions. The osteophytes were collected from knee joints of osteoarthritis patients (n=10, 94 osteophytes in total) after joint replacement surgery. Their size and origin in the respective joint were photo-documented. To develop an osteophyte classification, serial tissue sections were evaluated using histological (hematoxylin and eosin, Masson's trichrome, toluidine blue) and immunohistochemical staining (collagen type II). Based on the histological and immunohistochemical evaluation, osteophytes were categorized into four different types depending on the degree of ossification and the percentage of mesenchymal connective tissue. Size and localization of osteophytes were independent from the histological stages. This histological classification system of osteoarthritis osteophytes provides a helpful tool for analyzing and monitoring osteophyte development and for characterizing osteophyte types within a single human joint and may therefore contribute to achieve comparable results when analyzing histological findings in osteophytes. Copyright © 2015 Société française de rhumatologie. Published by Elsevier SAS. All rights reserved.

  14. 21 CFR 864.1850 - Dye and chemical solution stains.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical...

  15. Smoking habit and gastritis histology.

    Science.gov (United States)

    Namiot, A; Kemona, A; Namiot, Z

    2007-01-01

    Long-term cigarette smoking may increase the risk of digestive tract pathologies, however, what is the influence smoking habit on gastric mucosa histology is still poorly elicited. The aim of the study was to compare histological evaluation of gastritis in smoker and non-smoker groups. A total of 236 patients of various H. pylori status (109 infected, 127 non-infected), clinical diagnosis (107 duodenal ulcer disease, 129 dyspepsia), and smoking habit (92 smokers, 144 non-smokers) were included. Subjects were classified as smokers if they smoked 5 or more cigarettes per day for at least 3 years. A histological examination of endoscopically obtained samples was performed by two experienced pathomorphologists blinded to the diagnoses and smoking habit. Microscopic slices of the gastric mucosa were stained with hematoxylin-eosin and Giemsa. Apart from histological diagnosis, H. pylori status was additionally confirmed by an urease test (CLO-test) at least in one of two gastric locations (antrum or corpus). In the H. pylori infected population, H. pylori density, neutrophils, and mononuclear cells infiltration in the gastric corpus mucosa were lower in smokers than non-smokers, while in the antrum the differences were not significant. In the non-infected population, no significant differences in neutrophils and mononuclear cells infiltration between smokers and non-smokers were found. Since the significant differences in studied parameters of chronic gastritis between smokers and non-smokers were found in the corpus mucosa of H. pylori infected subjects, smoking should be taken into account when a histological evaluation of the gastric mucosa in the H. pylori infected population is performed.

  16. Treatment influencing down-staging in EORTC Melanoma Group sentinel node histological protocol compared with complete step-sectioning: a national multicentre study.

    Science.gov (United States)

    Riber-Hansen, Rikke; Hastrup, Nina; Clemmensen, Ole; Behrendt, Nille; Klausen, Siri; Ramsing, Mette; Spaun, Eva; Hamilton-Dutoit, Stephen Jacques; Steiniche, Torben

    2012-02-01

    Metastasis size in melanoma sentinel lymph nodes (SLNs) is an emerging prognostic factor. Two European melanoma treatment trials include SLN metastasis diameters as inclusion criteria. Whilst diameter estimates are sensitive to the number of sections examined, the level of this bias is largely unknown. We performed a prospective multicentre study to compare the European Organisation for Research and Treatment of Cancer (EORTC) recommended protocol with a protocol of complete step-sectioning. One hundred and thirty-three consecutive SLNs from seven SLN centres were analysed by five central sections 50μm apart (EORTC Protocol) followed by complete 250μm step-sectioning. Overall, 29 patients (21.8%) were SLN-positive. The EORTC Protocol missed eight of these metastases (28%), one metastasis measuring less than 0.1mm in diameter, seven measuring between 0.1 and 1mm. Complete step-sectioning at 250μm intervals (Extensive Protocol) missed one metastasis (3%) that measured less than 0.1mm. Thirteen treatment courses (34%) performed if inclusion was based on the Combined Protocol would not be performed if assessed by the EORTC Protocol. Thus, 10 patients would be without completion lymph node dissection (EORTC MINITUB study), whilst three patients would not be eligible for anti-CTLA4 trial (EORTC protocol 18071). The corresponding number with the Extensive Protocol would be three; one patient for the MINITUB registration study and two patients for the anti-CTLA4 study. Examining SLNs by close central sectioning alone (EORTC Protocol) misses a substantial number of metastases and underestimates the maximum metastasis diameter, leading to important changes in patient eligibility for various treatment protocols. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Two-photon excited fluorescence imaging of the pancreatic solid pseudopapillary tumor without hematoxylin and eosin stains.

    Science.gov (United States)

    Xu, Yahao; Liao, Chenxi; Chen, Jing; Chen, Youting; Zhu, Xiaoqin; Chen, Jianxin

    2016-05-01

    Solid pseudopapillary tumor (SPT) of the pancreas is an epithelial tumor with low-grade malignant potential and present more common in females. At present, the gold standard for accurate diagnosis of pancreatic tumor was mostly depending on the pathological and/or cytological evaluation. In this work, TPEF microscopy was applied to obtain the images of human normal pancreas and SPT of the pancreas without hematoxylin and eosin (H&E) staining, for the purpose of identifying the organization structural, cell morphological, and cytoplasm changing, which were then compared to their corresponding H&E stained histopathological results. Our results showed that high-resolution TPEF imaging of the pancreatic SPT can clearly distinguish the pathological features from normal pancreas in unstained histological sections, and the results are consistent with the histological results. Moreover, we measured the nuclear-cytoplasmic ratios of the pancreatic SPT and normal pancreas to characterize their difference in the cytomorphological feature. It indicated that this technique can achieve the consistent information of pathological diagnosis, and has the potential to substantially improve the optical diagnosis and treatment of the pancreatic SPT without H&E staining in the future. SCANNING 38:245-250, 2016. © 2015 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

  18. Toluidine Blue and Hematoxylin and Eosin Stains are Comparable in Evaluating Squamous Cell Carcinoma During Mohs.

    Science.gov (United States)

    Styperek, Andrew R; Goldberg, Leonard H; Goldschmidt, Laura E; Kimyai-Asadi, Arash

    2016-11-01

    Histologic examination of tissue is the foundation of Mohs micrographic surgery because determination of surgical margins influences whether additional tissue will be taken. Currently, there is no large focused study comparing toluidine blue (TB) and hematoxylin and eosin (H&E) stains in the evaluation of squamous cell carcinoma (SCC). This study evaluates whether TB and H&E are comparable in assessing the presence of tumor in frozen sections of SCC. One hundred eighty-six randomized slides representing 93 tissue pieces from 36 tumors were examined by 3 Mohs surgeons (1 Accreditation Council for Graduate Medical Education fellow and 2 fellowship-trained surgeons) and compared using a template that documented the presence and location of tumor on the slides. The evaluation of SCC with H&E and TB stains was highly concordant, with concordant identification of SCC in 96%, 96%, and 94% of tissue layers among the 3 Mohs surgeons ARS, LHG, and AK-A, respectively. Toluidine blue and H&E stains are statistically similar in their ability to detect SCC and guide Mohs surgical decision-making.

  19. A new technique for Gram staining paraffin-embedded tissue.

    Science.gov (United States)

    Engbaek, K; Johansen, K S; Jensen, M E

    1979-01-01

    Five techniques for Gram staining bacteria in paraffin sections were compared on serial sections of pulmonary tissues from eight bacteriological necropsies. Brown and Hopp's method was the most satisfactory for distinguishing Gram-positive and Gram-negative bacteria. However, this method cannot be recommended as the preparations were frequently overstained, and the Gram-negative bacteria were stained indistinctly. A modification of Brown and Hopps' method was developed which stains larger numbers of Gram-negative bacteria and differentiates well between different cell types and connective tissue, and there is no risk of overstaining. PMID:86548

  20. Calcification in calcium pyrophosphate dihydrate (CPPD) crystalline deposits in the knee: anatomic, radiographic, MR imaging, and histologic study in cadavers

    Energy Technology Data Exchange (ETDEWEB)

    Abreu, M.; Chung, C.B.; Lima, J.E. de; Trudell, D. [Department of Musculoskeletal Radiology, University of California, San Diego, VA San Diego Healthcare System, 3350 La Jolla Village Drive, CA 92162, San Diego (United States); Johnson, K.; Terkeltaub, R.; Resnick, D. [Department of Rheumatology, University of California, San Diego, VA San Diego Healthcare System, 3350 La Jolla Village Drive, CA 92162, San Diego (United States); Pe, S. [University of California, San Diego, VA San Diego Healthcare System, 3350 La Jolla Village Drive, CA 92162, San Diego (United States)

    2004-07-01

    To demonstrate and determine the frequency and location of calcification within cadaveric knees with or without calcification typical of calcium pyrophosphate dihydrate (CPPD), utilizing histologic, radiographic and MR imaging techniques. Ten cadaveric knees of elderly individuals that demonstrated no radiographic evidence of prior surgery or trauma were studied with MR imaging and subsequently sectioned in planes corresponding to those obtained with MR imaging. The slices were imaged with high-resolution radiography. Two musculoskeletal radiologists correlated the anatomic, MR and radiographic findings. Three of the knees, which did not demonstrate calcifications, were utilized as controls. Histologic sections were obtained from four knees that contained calcifications and from the three controls, and analyzed with special histologic stains that demonstrate phosphorus and calcium. Radiographic imaging and histologic analysis demonstrated widespread CPPD crystal deposition in four of the 10 knee specimens (40%). MR imaging demonstrated some calcifications only within the articular cartilage of the femoral condyles in three of the four (75%) specimens that had CPPD deposits. In all four specimens radiographs and histologic analysis were more sensitive than MR imaging. Histologic analysis demonstrated no evidence of CPPD crystals in the control specimens. MR imaging is insensitive to the presence of CPPD deposits in the knee, even when such deposits are widespread. Our study suggests that the sensitivity of MR imaging was significantly better in detecting CPPD deposits in the hyaline cartilage of the femoral condyles when compared with other internal structures, even when such structures contained a higher amount of calcification. (orig.)

  1. Facilitating tumor functional assessment by spatially relating 3D tumor histology and in vivo MRI: image registration approach.

    Directory of Open Access Journals (Sweden)

    Lejla Alic

    Full Text Available BACKGROUND: Magnetic resonance imaging (MRI, together with histology, is widely used to diagnose and to monitor treatment in oncology. Spatial correspondence between these modalities provides information about the ability of MRI to characterize cancerous tissue. However, registration is complicated by deformations during pathological processing, and differences in scale and information content. METHODOLOGY/PRINCIPAL FINDINGS: This study proposes a methodology for establishing an accurate 3D relation between histological sections and high resolution in vivo MRI tumor data. The key features of the methodology are: 1 standardized acquisition and processing, 2 use of an intermediate ex vivo MRI, 3 use of a reference cutting plane, 4 dense histological sampling, 5 elastic registration, and 6 use of complete 3D data sets. Five rat pancreatic tumors imaged by T2*-w MRI were used to evaluate the proposed methodology. The registration accuracy was assessed by root mean squared (RMS distances between manually annotated landmark points in both modalities. After elastic registration the average RMS distance decreased from 1.4 to 0.7 mm. The intermediate ex vivo MRI and the reference cutting plane shared by all three 3D images (in vivo MRI, ex vivo MRI, and 3D histology data were found to be crucial for the accurate co-registration between the 3D histological data set and in vivo MRI. The MR intensity in necrotic regions, as manually annotated in 3D histology, was significantly different from other histologically confirmed regions (i.e., viable and hemorrhagic. However, the viable and the hemorrhagic regions showed a large overlap in T2(*-w MRI signal intensity. CONCLUSIONS: The established 3D correspondence between tumor histology and in vivo MRI enables extraction of MRI characteristics for histologically confirmed regions. The proposed methodology allows the creation of a tumor database of spatially registered multi-spectral MR images and multi-stained 3D

  2. Histological study of white rhinoceros integument

    OpenAIRE

    Plochocki, Jeffrey H.; Ruiz, Saul; Rodriguez-Sosa, Jos? R.; Hall, Margaret I.

    2017-01-01

    In this study, we report findings from a microscopic analysis of the white rhinoceros (Ceratotherium simum) integumentary ultrastructure. Skin samples from the cheek, shoulder, flank and rump were taken from a 46-year-old female southern white rhinoceros and examined using H&E and elastic histological stains. The epidermis was thickest in the flank (1.003 mm) followed by the rump, cheek and shoulder. The stratum corneum comprised more than half the epidermal thickness. Numerous melanin granul...

  3. New Colors for Histology: Optimized Bivariate Color Maps Increase Perceptual Contrast in Histological Images.

    Science.gov (United States)

    Kather, Jakob Nikolas; Weis, Cleo-Aron; Marx, Alexander; Schuster, Alexander K; Schad, Lothar R; Zöllner, Frank Gerrit

    2015-01-01

    Accurate evaluation of immunostained histological images is required for reproducible research in many different areas and forms the basis of many clinical decisions. The quality and efficiency of histopathological evaluation is limited by the information content of a histological image, which is primarily encoded as perceivable contrast differences between objects in the image. However, the colors of chromogen and counterstain used for histological samples are not always optimally distinguishable, even under optimal conditions. In this study, we present a method to extract the bivariate color map inherent in a given histological image and to retrospectively optimize this color map. We use a novel, unsupervised approach based on color deconvolution and principal component analysis to show that the commonly used blue and brown color hues in Hematoxylin-3,3'-Diaminobenzidine (DAB) images are poorly suited for human observers. We then demonstrate that it is possible to construct improved color maps according to objective criteria and that these color maps can be used to digitally re-stain histological images. To validate whether this procedure improves distinguishability of objects and background in histological images, we re-stain phantom images and N = 596 large histological images of immunostained samples of human solid tumors. We show that perceptual contrast is improved by a factor of 2.56 in phantom images and up to a factor of 2.17 in sets of histological tumor images. Thus, we provide an objective and reliable approach to measure object distinguishability in a given histological image and to maximize visual information available to a human observer. This method could easily be incorporated in digital pathology image viewing systems to improve accuracy and efficiency in research and diagnostics.

  4. New Colors for Histology: Optimized Bivariate Color Maps Increase Perceptual Contrast in Histological Images.

    Directory of Open Access Journals (Sweden)

    Jakob Nikolas Kather

    Full Text Available Accurate evaluation of immunostained histological images is required for reproducible research in many different areas and forms the basis of many clinical decisions. The quality and efficiency of histopathological evaluation is limited by the information content of a histological image, which is primarily encoded as perceivable contrast differences between objects in the image. However, the colors of chromogen and counterstain used for histological samples are not always optimally distinguishable, even under optimal conditions.In this study, we present a method to extract the bivariate color map inherent in a given histological image and to retrospectively optimize this color map. We use a novel, unsupervised approach based on color deconvolution and principal component analysis to show that the commonly used blue and brown color hues in Hematoxylin-3,3'-Diaminobenzidine (DAB images are poorly suited for human observers. We then demonstrate that it is possible to construct improved color maps according to objective criteria and that these color maps can be used to digitally re-stain histological images.To validate whether this procedure improves distinguishability of objects and background in histological images, we re-stain phantom images and N = 596 large histological images of immunostained samples of human solid tumors. We show that perceptual contrast is improved by a factor of 2.56 in phantom images and up to a factor of 2.17 in sets of histological tumor images.Thus, we provide an objective and reliable approach to measure object distinguishability in a given histological image and to maximize visual information available to a human observer. This method could easily be incorporated in digital pathology image viewing systems to improve accuracy and efficiency in research and diagnostics.

  5. TEM validation of immunohistochemical staining prior to assessment of tumour angiogenesis by computerised image analysis

    International Nuclear Information System (INIS)

    Killingsworth, M.C.

    2002-01-01

    Full text: Counts of microvessel density (MVD) within solid tumours have been shown to be an independent predictor of outcome with higher counts generally associated with a worse prognosis. These assessments are commonly performed on immunoperoxidase stained (IPX) sections with antibodies to CD34, CD31 and Factor VIII-related antigen routinely used as vascular markers. Tumour vascular density is thought to reflect the demand the growing neoplasm is placing on its feeding blood supply. Vascular density also appears to be associated with spread of invasive cells to distant sites. The present study of tumour angiogenesis in prostate cancer specimens aims to assess new vessel growth in addition to MVD counts. The hypothesis being that an assessment which takes into account vascular migration and proliferation as well as the number of patent vessels present may have improved predictive power over assessments based on MVD counts alone. We are employing anti-CD34 stained IPX sections which are digitally photographed and assessed by a computerised image analysis system. Our aim is to develop parameters whereby tumour angiogenesis may be assessed at the light microscopic level and then correlated with existing histological methods of tumour assessment such as Gleason grading. In order to use IPX stained sections for angiogenic assessment validation and understanding of the anti-CD34 immunostaining pattern was necessary. This involved the following steps: i) Morphological assessment of angiogenic changes present in tumour blood vessels. Morphological changes in endothelial cells and pericytes indicative of angiogenic activation are generally below the level of resolution available with light microscopy. TEM examination revealed endothelial cell budding, pericyte retraction, basement membrane duplication and endothelial sprout formation in capillaries and venules surrounding tumour glands. This information assisted with the development of parameters by which IPX sections

  6. Hidradenocarcinoma: a histological and immunohistochemical study.

    Science.gov (United States)

    Ko, Christine J; Cochran, Alistair J; Eng, William; Binder, Scott W

    2006-11-01

    The diagnosis of hidradenocarcinoma is difficult due to a combination of factors including inconsistent nomenclature/ classification, rarity of the neoplasm, and variable morphology of cells composing the neoplasm. Immunohistochemistry has not been previously performed on a series of hidradenocarcinomas. We evaluated six cases of hidradenocarcinoma histologically and immunohistochemically using antibodies to gross cystic disease fluid protein-15 (GCDFP-15), carcino-embryonic antigen (CEA), epithelial membrane antigen (EMA), S-100 protein, keratin AE1/3, cytokeratin 5/6, p53, bcl-1, bcl-2, and Ki67. Histology suggested concurrent eccrine and apocrine differentiation of the cases. Ki67 and p53 staining was strongly positive in five of six tumors. The neoplasms stained with antibodies to CEA, S-100 protein, GCDFP-15, EMA, bcl-1, and bcl-2 in no consistent pattern. All tumors studied stained positively for keratin AE1/3 and cytokeratin 5/6. In making the diagnosis of hidradenocarcinoma, it may be unnecessary to separate hidradenocarcinoma into eccrine and apocrine categories, and although Ki67 and p53 may be helpful, histological parameters remain paramount.

  7. Histological analysis of thelohaniasis in white-clawed crayfish Austropotamobius pallipes complex

    Directory of Open Access Journals (Sweden)

    Quaglio F.

    2011-09-01

    Full Text Available From 2004 to 2006, a parasitological survey aimed at the detection of the microsporidian parasite Thelohania contejeani Henneguy was carried out on 177 wild white-clawed crayfish (Austropotamobius pallipes complex captured in six streams and rivers of the province of Belluno in north-eastern Italy. Microscopical examination of the skeletal muscles, and histological analysis applying different histochemical stains to full transverse and sagittal sections of the cephalothorax and abdomen were carried out. Transmission electron microscopy (TEM was also conducted on the parasites recovered during the survey. Out of 177 crayfish examined, Thelohania contejeani (Microsporidia, Thelohaniidae was present in only one crayfish from the Vena d’oro creek. The parasite was detected in the skeletal muscles in several developmental stages, including mature spores, which represented the most common stage recovered. Sporophorous vesicles were also present. Histological examination revealed that the fibres of the skeletal, cardiac and intestinal muscles were filled with spores. Melanin infiltrations were focally present in the infected striated muscles. The gill phagocytic nephrocytes were engulfed by small masses of spores. Among the staining techniques applied, Crossman’s trichrome stain represented the most effective method of detecting T. contejeani.

  8. Multiparametric MRI of Epiphyseal Cartilage Necrosis (Osteochondrosis with Histological Validation in a Goat Model.

    Directory of Open Access Journals (Sweden)

    Luning Wang

    Full Text Available To evaluate multiple MRI parameters in a surgical model of osteochondrosis (OC in goats.Focal ischemic lesions of two different sizes were induced in the epiphyseal cartilage of the medial femoral condyles of goats at 4 days of age by surgical transection of cartilage canal blood vessels. Goats were euthanized and specimens harvested 3, 4, 5, 6, 9 and 10 weeks post-op. Ex vivo MRI scans were conducted at 9.4 Tesla for mapping the T1, T2, T1ρ, adiabatic T1ρ and TRAFF relaxation times of articular cartilage, unaffected epiphyseal cartilage, and epiphyseal cartilage within the area of the induced lesion. After MRI scans, safranin O staining was conducted to validate areas of ischemic necrosis induced in the medial femoral condyles of six goats, and to allow comparison of MRI findings with the semi-quantitative proteoglycan assessment in corresponding safranin O-stained histological sections.All relaxation time constants differentiated normal epiphyseal cartilage from lesions of ischemic cartilage necrosis, and the histological staining results confirmed the proteoglycan (PG loss in the areas of ischemia. In the scanned specimens, all of the measured relaxation time constants were higher in the articular than in the normal epiphyseal cartilage, consistently allowing differentiation between these two tissues.Multiparametric MRI provided a sensitive approach to discriminate between necrotic and viable epiphyseal cartilage and between articular and epiphyseal cartilage, which may be useful for diagnosing and monitoring OC lesions and, potentially, for assessing effectiveness of treatment interventions.

  9. Using color and grayscale images to teach histology to color-deficient medical students.

    Science.gov (United States)

    Rubin, Lindsay R; Lackey, Wendy L; Kennedy, Frances A; Stephenson, Robert B

    2009-01-01

    Examination of histologic and histopathologic microscopic sections relies upon differential colors provided by staining techniques, such as hematoxylin and eosin, to delineate normal tissue components and to identify pathologic alterations in these components. Given the prevalence of color deficiency (commonly called "color blindness") in the general population, it is likely that this reliance upon color differentiation poses a significant obstacle for several medical students beginning a course of study that includes examination of histologic slides. In the past, first-year medical students at Michigan State University who identified themselves as color deficient were encouraged to use color transparency overlays or tinted contact lenses to filter out problematic colors. Recently, however, we have offered such students a computer monitor adjusted to grayscale for in-lab work, as well as grayscale copies of color photomicrographs for examination purposes. Grayscale images emphasize the texture of tissues and the contrasts between tissues as the students learn histologic architecture. Using this approach, color-deficient students have quickly learned to compensate for their deficiency by focusing on cell and tissue structure rather than on color variation. Based upon our experience with color-deficient students, we believe that grayscale photomicrographs may also prove instructional for students with normal (trichromatic) color vision, by encouraging them to consider structural characteristics of cells and tissues that may otherwise be overshadowed by stain colors.

  10. Enhanced Visualization of Hematoxylin and Eosin Stained Pathological Characteristics by Phasor Approach.

    Science.gov (United States)

    Luo, Teng; Lu, Yuan; Liu, Shaoxiong; Lin, Danying; Qu, Junle

    2017-09-05

    The phasor approach to fluorescence lifetime imaging microscopy (FLIM) is used to identify different types of tissues from hematoxylin and eosin (H&E) stained basal cell carcinoma (BCC) sections. The results suggest that working directly on the phasor space with the clustering assignment achieves immunofluorescence like simultaneous five or six-color imaging by using multiplexed fluorescence lifetimes of H&E. The phase approach is of particular effectiveness for enhanced visualization of the abnormal morphology of a suspected nidus. Moreover, the phasor approach to H&E FLIM data can determine the actual paths or the infiltrating trajectories of basophils and immune cells associated with the preneoplastic or neoplastic skin lesions. The integration of the phasor approach with routine histology proved its available value for skin cancer prevention and early detection. We therefore believe that the phasor analysis of H&E tissue sections is an enhanced visualization tool with the potential to simplify the preparation process of special staining and serve as color contrast aided imaging in clinical pathological examination.

  11. Automated Analysis and Classification of Histological Tissue Features by Multi-Dimensional Microscopic Molecular Profiling.

    Directory of Open Access Journals (Sweden)

    Daniel P Riordan

    Full Text Available Characterization of the molecular attributes and spatial arrangements of cells and features within complex human tissues provides a critical basis for understanding processes involved in development and disease. Moreover, the ability to automate steps in the analysis and interpretation of histological images that currently require manual inspection by pathologists could revolutionize medical diagnostics. Toward this end, we developed a new imaging approach called multidimensional microscopic molecular profiling (MMMP that can measure several independent molecular properties in situ at subcellular resolution for the same tissue specimen. MMMP involves repeated cycles of antibody or histochemical staining, imaging, and signal removal, which ultimately can generate information analogous to a multidimensional flow cytometry analysis on intact tissue sections. We performed a MMMP analysis on a tissue microarray containing a diverse set of 102 human tissues using a panel of 15 informative antibody and 5 histochemical stains plus DAPI. Large-scale unsupervised analysis of MMMP data, and visualization of the resulting classifications, identified molecular profiles that were associated with functional tissue features. We then directly annotated H&E images from this MMMP series such that canonical histological features of interest (e.g. blood vessels, epithelium, red blood cells were individually labeled. By integrating image annotation data, we identified molecular signatures that were associated with specific histological annotations and we developed statistical models for automatically classifying these features. The classification accuracy for automated histology labeling was objectively evaluated using a cross-validation strategy, and significant accuracy (with a median per-pixel rate of 77% per feature from 15 annotated samples for de novo feature prediction was obtained. These results suggest that high-dimensional profiling may advance the

  12. HistologiQuiz

    DEFF Research Database (Denmark)

    Brent, Mikkel Bo

    2015-01-01

    HistologiQuiz er en quiz-app udviklet til almen og speciel histologi. Den består af mere end 1400 spørgsmål og over 320 histologiske billeder. Alle spørgsmål tager udgangspunkt i lærebogen Genesers Histologi af Annemarie Brüel m.fl.......HistologiQuiz er en quiz-app udviklet til almen og speciel histologi. Den består af mere end 1400 spørgsmål og over 320 histologiske billeder. Alle spørgsmål tager udgangspunkt i lærebogen Genesers Histologi af Annemarie Brüel m.fl....

  13. Infrared spectral histopathology using haematoxylin and eosin (H&E) stained glass slides: a major step forward towards clinical translation.

    Science.gov (United States)

    Pilling, Michael J; Henderson, Alex; Shanks, Jonathan H; Brown, Michael D; Clarke, Noel W; Gardner, Peter

    2017-04-10

    Infrared spectral histopathology has shown great promise as an important diagnostic tool, with the potential to complement current pathological methods. While promising, clinical translation has been hindered by the impracticalities of using infrared transmissive substrates which are both fragile and prohibitively very expensive. Recently, glass has been proposed as a potential replacement which, although largely opaque in the infrared, allows unrestricted access to the high wavenumber region (2500-3800 cm -1 ). Recent studies using unstained tissue on glass have shown that despite utilising only the amide A band, good discrimination between histological classes could be achieved, and suggest the potential of discriminating between normal and malignant tissue. However unstained tissue on glass has the potential to disrupt the pathologist workflow, since it needs to be stained following infrared chemical imaging. In light of this, we report on the very first infrared Spectral Histopathology SHP study utilising coverslipped H&E stained tissue on glass using samples as received from the pathologist. In this paper we present a rigorous study using results obtained from an extended patient sample set consisting of 182 prostate tissue cores obtained from 100 different patients, on 18 separate H&E slides. Utilising a Random Forest classification model we demonstrate that we can rapidly classify four classes of histology of an independent test set with a high degree of accuracy (>90%). We investigate different degrees of staining using nine separate prostate serial sections, and demonstrate that we discriminate on biomarkers rather than the presence of the stain. Finally, using a four-class model we show that we can discriminate normal epithelium, malignant epithelium, normal stroma and cancer associated stroma with classification accuracies over 95%.

  14. Clinico-Histologic Conferences: Histology and Disease

    Science.gov (United States)

    Shaw, Phyllis A.; Friedman, Erica S.

    2012-01-01

    Providing a context for learning information and requiring learners to teach specific content has been demonstrated to enhance knowledge retention. To enhance students' appreciation of the role of science and specifically histology in clinical reasoning, disease diagnosis, and treatment, a new teaching format was created to provide clinical…

  15. Measurement of neuron soma size by fluorescent Nissl stain

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: James Cronk, Noel Derecki & Jonathan Kipnis ### Abstract This protocol describes how to measure neuron soma size by fluorescent Nissl stain. Mice are sacrificed, and fixed by PFA perfusion. Brains are removed, and further PFA fixed, followed by sucrose cryoprotection. They are then snap frozen, sliced by cryostat, and stained with fluorescent Nissl as floating sections. Confocal microscopy is used to take images of neurons, and a computer graphics tablet is used to calculate ...

  16. Real-time histology in liver disease using multiphoton microscopy with fluorescence lifetime imaging

    OpenAIRE

    Wang, Haolu; Liang, Xiaowen; Mohammed, Yousuf H.; Thomas, James A.; Bridle, Kim R.; Thorling, Camilla A.; Grice, Jeffrey E.; Xu, Zhi Ping; Liu, Xin; Crawford, Darrell H. G.; Roberts, Michael S.

    2015-01-01

    Conventional histology with light microscopy is essential in the diagnosis of most liver diseases. Recently, a concept of real-time histology with optical biopsy has been advocated. In this study, live mice livers (normal, with fibrosis, steatosis, hepatocellular carcinoma and ischemia-reperfusion injury) were imaged by MPM-FLIM for stain-free real-time histology. The acquired MPM-FLIM images were compared with conventional histological images. MPM-FLIM imaged subsurface cellular and subcellu...

  17. Nuclear staining with alum hematoxylin.

    Science.gov (United States)

    Llewellyn, B D

    2009-08-01

    The hematoxylin and eosin stain is the most common method used in anatomic pathology, yet it is a method about which technologists ask numerous questions. Hematoxylin is a natural dye obtained from a tree originally found in Central America, and is easily converted into the dye hematein. This dye forms coordination compounds with mordant metals, such as aluminum, and the resulting lake attaches to cell nuclei. Regressive formulations contain a higher concentration of dye than progressive formulations and may also contain a lower concentration of mordant. The presence of an acid increases the life of the solution and in progressive solutions may also affect selectivity of staining. An appendix lists more than 60 hemalum formulations and the ratio of dye to mordant for each.

  18. Etika Berbusana Mahasiswa Stain Samarinda

    Directory of Open Access Journals (Sweden)

    Ida Suryani Wijaya

    2012-06-01

    Full Text Available Ethics is about behavior of human being, such as which one is right or wrong. The ethics is always affecting the human life. The ethics gives people orientation how he/she do manything every time every day. Islamic ethics consists of the way how someone interact each other; how someone should do or not to do, how to sit, how to walk, how to eat or drink, how to sleep, or how to get dressed. Al-Qur’an uses three terms to define about dressing, they are: libas, tsiyah, and sarahi. Dressing has a function as covering the body, as assessoris, as the way to do Islamic taqwa, and as an identiy. Dressing ethics of the female students of STAIN Samarinda has been regulated by the rector regulation No 19 of the year 2002 about relation and dressing ethics for the students of STAIN Samarinda.

  19. Mapping stain distribution in pathology slides using whole slide imaging

    Directory of Open Access Journals (Sweden)

    Fang-Cheng Yeh

    2014-01-01

    Full Text Available Background: Whole slide imaging (WSI offers a novel approach to digitize and review pathology slides, but the voluminous data generated by this technology demand new computational methods for image analysis. Materials and Methods: In this study, we report a method that recognizes stains in WSI data and uses kernel density estimator to calculate the stain density across the digitized pathology slides. The validation study was conducted using a rat model of acute cardiac allograft rejection and another rat model of heart ischemia/reperfusion injury. Immunohistochemistry (IHC was conducted to label ED1 + macrophages in the tissue sections and the stained slides were digitized by a whole slide scanner. The whole slide images were tessellated to enable parallel processing. Pixel-wise stain classification was conducted to classify the IHC stains from those of the background and the density distribution of the identified IHC stains was then calculated by the kernel density estimator. Results: The regression analysis showed a correlation coefficient of 0.8961 between the number of IHC stains counted by our stain recognition algorithm and that by the manual counting, suggesting that our stain recognition algorithm was in good agreement with the manual counting. The density distribution of the IHC stains showed a consistent pattern with those of the cellular magnetic resonance (MR images that detected macrophages labeled by ultrasmall superparamagnetic iron-oxide or micron-sized iron-oxide particles. Conclusions: Our method provides a new imaging modality to facilitate clinical diagnosis. It also provides a way to validate/correlate cellular MRI data used for tracking immune-cell infiltration in cardiac transplant rejection and cardiac ischemic injury.

  20. Tc-99m annexin V imaging and apoptosis staining in rabbit model of osteoarthritis

    International Nuclear Information System (INIS)

    Kang, Do Young; Jeong, Young Jin; Choi, Sun Mee; Lee, Sung Won; Chung, Won Tae; Yoo, Young Hyun

    2004-01-01

    Recent studies showed that, in osteoarthritis (OA), articular chondrocytes appeared to be eliminated by apoptosis. Tc-99mm Annexin V has been successfully used for non-invasive gamma imaging of apoptosis in tumor. myocardial infarction and transplantation. We studied Tc-99m Annexin V imaging and apoptosis staining in rabbit model of osteoarthritis. Osteoarthritis was induced in rabbits by intra-articular injection of 1.0 mg collagenase and surgical transection of leg ligament. Animals were dissected at 2, 4, 6 and 8 weeks after the initiation of the injections. For histological observation, the paraffin sections were stained with hematoxylin and eosin. To confirm that osteoarthritis was induced, immunohistochemistry to TRAIL was conducted on the sections and TRAIL positive cells was revealed by DAB. Tc-99m Annexin V was injected 16 ug/1 mCi/kg and regional images were acquired 10 and 60 min postinjection. A few days later Tc-99m MDP imaging was also acquired. Two weeks after injection, the surface layer of cartilage was lost, chondrocytes in the transitional zone have disappeared, and cleft in the transitional zone were shown. Four weeks after injection, moderate cell cloning in transitional and radial zone was appeared. Six weeks after injection, cell cloning was more apparent in the transitional and radial zones. Whereas TRAIL positive cells were not found in the chondrocytes from the control cartilage, most chondrocytes from the collagenase injected cartilage were TRAIL-positive. Tc-99m Annexin V imaging showed increase uptake in knee area of injuried side to normal side. And Tc-99m MDP imaging also had same findings. In rabbit model of osteoarthritis, apoptosis was detected by Tc-99m Annexin V imaging noninvasively and it was correlated by pathologic staining for apoptosis

  1. Spectral CT of carotid atherosclerotic plaque: comparison with histology

    Energy Technology Data Exchange (ETDEWEB)

    Zainon, R.; Doesburg, R.M. [University of Canterbury, Department of Physics and Astronomy, Christchurch (New Zealand); Ronaldson, J.P.; Gieseg, S.P. [University of Otago, Centre for Bioengineering, Christchurch (New Zealand); Janmale, T. [University of Canterbury, Free Radical Biochemistry Laboratory, School of Biological Sciences, Christchurch (New Zealand); Scott, N.J. [University of Otago, Department of Medicine, Christchurch (New Zealand); Buckenham, T.M. [University of Otago, Department of Academic Radiology, Christchurch (New Zealand); Butler, A.P.H. [University of Otago, Centre for Bioengineering, Christchurch (New Zealand); University of Otago, Department of Academic Radiology, Christchurch (New Zealand); University of Canterbury, Department of Electrical and Computer Engineering, Christchurch (New Zealand); European Organisation for Nuclear Research (CERN), Geneva (Switzerland); Butler, P.H. [University of Canterbury, Department of Physics and Astronomy, Christchurch (New Zealand); European Organisation for Nuclear Research (CERN), Geneva (Switzerland); Roake, J.A. [Christchurch Hospital, Department of Vascular, Endovascular and Transplant Surgery, Christchurch (New Zealand); Anderson, N.G. [University of Otago, Centre for Bioengineering, Christchurch (New Zealand); University of Otago, Department of Academic Radiology, Christchurch (New Zealand); University of Otago, Christchurch, Department of Radiology, PO Box 4345, Christchurch (New Zealand)

    2012-12-15

    To distinguish components of vulnerable atherosclerotic plaque by imaging their energy response using spectral CT and comparing images with histology. After spectroscopic calibration using phantoms of plaque surrogates, excised human carotid atherosclerotic plaques were imaged using MARS CT using a photon-processing detector with a silicon sensor layer and microfocus X-ray tube (50 kVp, 0.5 mA) at 38-{mu}m voxel size. The plaques were imaged, sectioned and re-imaged using four threshold energies: 10, 16, 22 and 28 keV; then sequentially stained with modified Von Kossa, Perl's Prussian blue and Oil-Red O, and photographed. Relative Hounsfield units across the energies were entered into a linear algebraic material decomposition model to identify the unknown plaque components. Lipid, calcium, iron and water-like components of plaque have distinguishable energy responses to X-ray, visible on spectral CT images. CT images of the plaque surface correlated very well with histological photographs. Calcium deposits (>1,000 {mu}m) in plaque are larger than iron deposits (<100 {mu}m), but could not be distinguished from each other within the same voxel using the energy range available. Spectral CT displays energy information in image form at high spatial resolution, enhancing the intrinsic contrast of lipid, calcium and iron within atheroma. (orig.)

  2. Distinguishing the Chinese materia medica Tiepishihu from similar Dendrobium species of the same genus using histological and microscopic method.

    Science.gov (United States)

    Yu, Kun-Zi; Yan, Hua; Tai, Hai-Chuan; Zhang, Nan-Ping; Cheng, Xian-Long; Guo, Zeng-Xi; Ma, Shuang-Cheng; Wei, Feng

    2017-07-01

    The Chinese Materia Medica, Tiepishihu, used as a tonic for over one thousand years, is a well-known precious medicine in China. According to the Chinese Pharmacopoeia, its source is the species Dendrobium officinale Kimura et Migo, which is distinguished from other species in Dendrobium genus. However, these species from the same genus are similar with Tiepishihu and caused confusion in the market. To find a quick and simple method to distinguish Tiepishihu from other similar species, histologic and microscopic methods were combined together to investigate the transverse section of stem of Tiepishihu and other similar species. Phloroglucinol test solution with hydrochloric acid was used to reveal the lignified tissue by staining the transverse section of Tiepishihu and similar species. Results revealed the unique identification characteristics to distinguish Tiepishihu from similar species, which were difficult to distinguish by other methods. The identification characteristics of Tiepishihu include the cells of vascular bundle sheath were stained red, parenchyma cells were not stained red. What's more, other species can be distinguished from each other with microscopic and histological characteristics. These characteristics proved stable and can be easily observed by normal light microscopic examination. This method is rapid, accurate, stable, and inexpensive. © 2017 Wiley Periodicals, Inc.

  3. LANTHANUM STAINING OF THE SURFACE COAT OF CELLS

    Science.gov (United States)

    Shea, Stephen M.

    1971-01-01

    Among the techniques which have been reported to stain the surface coat of cells, for electron microscopy, is lanthanum staining en bloc. Similarly, the presence of the cationic dye, Alcian blue 8GX, in a primary glutaraldehyde fixative has been reported to improve the preservation of the surface coat of cells of many types; however, the preserved coat is not very electron opaque unless thin sections are counterstained. The present paper shows that for several rat tissues lanthanum staining en bloc is an effective electron stain for the cell surface, giving excellent contrast, if combined sequentially with prefixation in an aldehyde fixative containing Alcian blue. The cationic substance cetylpyridinium chloride was found to have a similar effect to that of Alcian blue in enhancing the lanthanum staining of the surface coat material of the brush border of intestinal epithelial cells. The patterns of lanthanum staining obtained for the tissues studied strikingly resemble those reported in the literature where tissues are stained by several standard methods for demonstrating mucosubstances at the ultrastructural level. This fact and the reproduction of the effect of Alcian blue by cetylpyridinium chloride constitute a persuasive empirical argument that the material visualized is a mucopolysaccharide or mucopolysaccharide-protein complex. PMID:4108476

  4. Matrix Remodeling During Intervertebral Disc Growth and Degeneration Detected by Multichromatic FAST Staining

    Science.gov (United States)

    Leung, Victor Y.L.; Chan, Wilson C.W.; Hung, Siu-Chun; Cheung, Kenneth M.C.; Chan, Danny

    2009-01-01

    Various imaging techniques have been used to assess degeneration of the intervertebral disc, including many histological methods, but cartilage-oriented histological stains do not clearly show the comparatively complex structures of the disc. In addition, there is no integrated method to assess efficiently both the compartmental organization and matrix composition in disc samples. In this study, a novel histological method, termed FAST staining, has been developed to investigate disc growth and degeneration by sequential staining with fast green, Alcian blue, Safranin-O, and tartrazine to generate multichromatic histological profiles (FAST profiles). This identifies the major compartments of the vertebra-disc region, including the cartilaginous endplate and multiple zones of the annulus fibrosus, by specific FAST profile patterns. A disc degeneration model in rabbit established using a previously described puncture method showed gradual but profound alteration of the FAST profile during disc degeneration, supporting continual alteration of glycosaminoglycan. Changes of the FAST profile pattern in the nucleus pulposus and annulus fibrosus of the postnatal mouse spine suggested matrix remodeling activity during the growth of intervertebral discs. In summary, we developed an effective staining method capable of defining intervertebral disc compartments in detail and showing matrix remodeling events within the disc. The FAST staining method may be used to develop a histopathological grading system to evaluate disc degeneration or malformation. (J Histochem Cytochem 57:249–256, 2009) PMID:19001641

  5. Myxosporidiosis in intensively-reared Piaractus mesopotamicus: Histopathological diagnosis by means of Ziehl-Neelsen staining

    Directory of Open Access Journals (Sweden)

    Wilson G. Manrique

    2012-11-01

    Full Text Available Samples of different organs from intensively-reared Piaractus mesopotamicus were collected and processed using routine histological techniques in order to produce thin sections for staining with hematoxylin-eosin and with the Ziehl-Neelsen method. Through examination under an optical microscope, myxosporidians of the genera Henneguya sp. and Myxobolus sp. were identified, respectivelyin the gills and kidneys of P. mesopotamicus. Plasmodia with immature spores of Henneguya sp. were located along the secondary lamellae, with total length of 30.45±4.84µm and width of 3.52±0.33µm. Spores of Myxobolus sp. were located in the kidneys, with total length of 8.94±0.82µm and width of 5.59±0.39µm. Histopathological analysis of the gills showed plasmodia containing spores of Henneguya sp., at intralamellar and intravascular localities, at different stages of development. Spores of Myxobolus sp. were identified in the kidneys, in the peritubular region and in the interstices and glomerulus, surrounded by melanomacrophages. Focal hemorrhage was recorded in a few cases. Ziehl-Neelsen staining allowed to identify particular features of the spores and facilitated biometry and enabled classification in comparison with hematoxylin-eosin, thus demonstrating its usefulness for histopathological diagnosis of the parasitosis.

  6. The influence of external ultrasound on the histologic architecture of the organic capsule around smooth silicone implants: experimental study in rats.

    Science.gov (United States)

    Mendes, F H; Viterbo, F; DeLucca, L

    2008-05-01

    Capsular contracture is the main complication related to breast silicone implants, and its prevention remains a medical challenge. The authors present experimental research examining the effect of external ultrasound on the formation and contracture of peri-implant capsules. In this study, 42 male Wistar rats had a 2-mm smooth surface implant placed in a dorsal submuscular pocket. They then were separated into "ultrasound" and "control" groups that received repeated external applications either with or without the ultrasound power on. Ultrasound applications were given three times a week for a period of 90 days. After that, both groups were housed under the same conditions with no application scheduled. Five animals of each group, killed at 30, 60, 90, and 180 days, had their implants removed along with the capsule, which received a special histologic preparation via annular sectioning that provided wide circumferential observation of the capsular tissue. Sections were stained with hematoxylin/eosin stain, Masson's trichrome stain, and Pricrosirius Red stain for regular microscopic evaluation under normal and polarized light. Histologic data showed that capsules from the ultrasound and control groups had statistically significant differences. Ultrasound application developed a capsular architecture similar to that shown within textured silicone implants, and its effect had an early definition with subsequent stabilization. The authors conclude that early and repeated external ultrasound application enhances the thickness, cellular count, and vascularity of smooth silicone capsular tissue, whereas it diminishes the pattern of parallel orientation of collagen fibers.

  7. Prognostic Utility of Histological Growth Patterns of Colorectal Lung Oligometastasis.

    Science.gov (United States)

    Yeong, Son Jae; Pak, Min Gyoung; Lee, Hyoun Wook; Ha, Seung Yeon; Roh, Mee Sook

    2018-03-01

    Patients with resectable colorectal lung oligometastasis (CLOM) demonstrate a heterogeneous oncological outcome. However, the parameters for predicting tumor aggressiveness have not yet been fully investigated in CLOM. This study was performed to determine the prognostic value of histological growth patterns in patients who underwent surgery for CLOM. The study included 92 patients who were diagnosed with CLOM among the first resection cases. CLOMs grow according to three histological patterns: aerogenous, pushing, and desmoplastic patterns. The growth patterns were evaluated on archival hematoxylin and eosin-stained tissue sections. The aerogenous pattern was found in 29.4% (n=27) of patients, the pushing pattern in 34.7% (n=32), the desmoplastic pattern in 6.5% (n=6), and a mix of two growth patterns in 29.4% (n=27). The size of the aerogenous pattern was significantly smaller than that of metastases with other patterns (p=.033). Kaplan-Meier analysis demonstrated that patients showing an aerogenous pattern appeared to have a poorer prognosis, which was calculated from the time of diagnosis of the CLOM (p=.044). The 5-year survival rate from the diagnosis of colorectal cancer tended to be lower in patients with an aerogenous pattern than in those who had a non-aerogenous pattern; however, the difference was marginally significant (p=.051). In the multivariate Cox analysis, the aerogenous pattern appeared as an independent predictor of poor overall survival (hazard ratio, 3.122; 95% confidence interval, 1.196 to 8.145; p=.020). These results suggest that the growth patterns may play a part as a histology-based prognostic parameter for patients with CLOM.

  8. Prognostic Utility of Histological Growth Patterns of Colorectal Lung Oligometastasis

    Directory of Open Access Journals (Sweden)

    Son Jae Yeong

    2018-03-01

    Full Text Available Background Patients with resectable colorectal lung oligometastasis (CLOM demonstrate a heterogeneous oncological outcome. However, the parameters for predicting tumor aggressiveness have not yet been fully investigated in CLOM. This study was performed to determine the prognostic value of histological growth patterns in patients who underwent surgery for CLOM. Methods The study included 92 patients who were diagnosed with CLOM among the first resection cases. CLOMs grow according to three histological patterns: aerogenous, pushing, and desmoplastic patterns. The growth patterns were evaluated on archival hematoxylin and eosin–stained tissue sections. Results The aerogenous pattern was found in 29.4% (n=27 of patients, the pushing pattern in 34.7% (n=32, the desmoplastic pattern in 6.5% (n=6, and a mix of two growth patterns in 29.4% (n=27. The size of the aerogenous pattern was significantly smaller than that of metastases with other patterns (p=.033. Kaplan-Meier analysis demonstrated that patients showing an aerogenous pattern appeared to have a poorer prognosis, which was calculated from the time of diagnosis of the CLOM (p=.044. The 5-year survival rate from the diagnosis of colorectal cancer tended to be lower in patients with an aerogenous pattern than in those who had a non-aerogenous pattern; however, the difference was marginally significant (p=.051. In the multivariate Cox analysis, the aerogenous pattern appeared as an independent predictor of poor overall survival (hazard ratio, 3.122; 95% confidence interval, 1.196 to 8.145; p=.020. Conclusions These results suggest that the growth patterns may play a part as a histology-based prognostic parameter for patients with CLOM.

  9. Color normalization of histology slides using graph regularized sparse NMF

    Science.gov (United States)

    Sha, Lingdao; Schonfeld, Dan; Sethi, Amit

    2017-03-01

    Computer based automatic medical image processing and quantification are becoming popular in digital pathology. However, preparation of histology slides can vary widely due to differences in staining equipment, procedures and reagents, which can reduce the accuracy of algorithms that analyze their color and texture information. To re- duce the unwanted color variations, various supervised and unsupervised color normalization methods have been proposed. Compared with supervised color normalization methods, unsupervised color normalization methods have advantages of time and cost efficient and universal applicability. Most of the unsupervised color normaliza- tion methods for histology are based on stain separation. Based on the fact that stain concentration cannot be negative and different parts of the tissue absorb different stains, nonnegative matrix factorization (NMF), and particular its sparse version (SNMF), are good candidates for stain separation. However, most of the existing unsupervised color normalization method like PCA, ICA, NMF and SNMF fail to consider important information about sparse manifolds that its pixels occupy, which could potentially result in loss of texture information during color normalization. Manifold learning methods like Graph Laplacian have proven to be very effective in interpreting high-dimensional data. In this paper, we propose a novel unsupervised stain separation method called graph regularized sparse nonnegative matrix factorization (GSNMF). By considering the sparse prior of stain concentration together with manifold information from high-dimensional image data, our method shows better performance in stain color deconvolution than existing unsupervised color deconvolution methods, especially in keeping connected texture information. To utilized the texture information, we construct a nearest neighbor graph between pixels within a spatial area of an image based on their distances using heat kernal in lαβ space. The

  10. A novel histological technique for distinguishing between epithelial cells in forensic casework.

    Science.gov (United States)

    French, Claire E V; Jensen, Cynthia G; Vintiner, Susan K; Elliot, Douglas A; McGlashan, Susan R

    2008-06-10

    There are a number of forensic cases in which the identification of the epithelial cell type from which DNA originated would provide important probative evidence. This study aimed to develop a technique using histological staining of fixed cells to distinguish between skin, buccal and vaginal epithelium. First, 11 different stains were screened on formalin-fixed, wax-embedded cells from five women. Samples were analysed qualitatively by examining staining patterns (colour) and morphology (absence or presence of nuclei). Three of the staining methods--Dane's, Csaba's and Ayoub-Shklar--were successful in distinguishing skin epithelial cells from buccal and vaginal. Second, cells were smeared directly onto slides, fixed with one of five fixatives and stained with one of the three stains mentioned above. Methanol fixation, coupled with the Dane's staining method, specific to keratin, was the only technique that distinguished between all three cell types. Skin cells stained magenta, red and orange and lacked nuclei; buccal cells stained predominantly orange-pink with red nuclei; while vaginal cells stained bright orange with orange nuclei and a blue extracellular hue. This staining pattern in vaginal cells was consistent in samples collected from 50 women aged between 18 and 67. Identification of cell type from unlabelled micrographs by 10 trained observers showed a mean success rate of 95%. The results of this study demonstrate that histological staining may provide forensic scientists with a technique for distinguishing between skin, buccal and vaginal epithelial cells and thus would enable more conclusive analyses when investigating sexual assault cases.

  11. Digital simulation of staining in histopathology multispectral images: enhancement and linear transformation of spectral transmittance.

    Science.gov (United States)

    Bautista, Pinky A; Yagi, Yukako

    2012-05-01

    Hematoxylin and eosin (H&E) stain is currently the most popular for routine histopathology staining. Special and/or immuno-histochemical (IHC) staining is often requested to further corroborate the initial diagnosis on H&E stained tissue sections. Digital simulation of staining (or digital staining) can be a very valuable tool to produce the desired stained images from the H&E stained tissue sections instantaneously. We present an approach to digital staining of histopathology multispectral images by combining the effects of spectral enhancement and spectral transformation. Spectral enhancement is accomplished by shifting the N-band original spectrum of the multispectral pixel with the weighted difference between the pixel's original and estimated spectrum; the spectrum is estimated using M transformed to the spectral configuration associated to its reaction to a specific stain by utilizing an N × N transformation matrix, which is derived through application of least mean squares method to the enhanced and target spectral transmittance samples of the different tissue components found in the image. Results of our experiments on the digital conversion of an H&E stained multispectral image to its Masson's trichrome stained equivalent show the viability of the method.

  12. Integration of ultra-high field MRI and histology for connectome based research of brain disorders

    Directory of Open Access Journals (Sweden)

    Shan eYang

    2013-09-01

    Full Text Available Ultra-high field magnetic resonance imaging (MRI became increasingly relevant for in vivo neuroscientific research because of improved spatial resolutions. However, this is still the unchallenged domain of histological studies, which long played an important role in the investigation of neuropsychiatric disorders. While the field of biological psychiatry strongly advanced on macroscopic levels, current developments are rediscovering the richness of immunohistological information when attempting a multi-level systematic approach to brain function and dysfunction. For most studies, histology sections lost information on three-dimensional reconstructions. Translating histological sections to 3D-volumes would thus not only allow for multi-stain and multi-subject alignment in post mortem data, but also provide a crucial step in big data initiatives involving the network analyses currently performed with in vivo MRI. We therefore investigated potential pitfalls during integration of MR and histological information where no additional blockface information is available. We demonstrated that strengths and requirements from both methods seem to be ideally merged at a spatial resolution of 200 μm. However, the success of this approach is heavily dependent on choices of hardware, sequence and reconstruction. We provide a fully automated pipeline that optimizes histological 3D reconstructions, providing a potentially powerful solution not only for primary human post mortem research institutions in neuropsychiatric research, but also to help alleviate the massive workloads in neuroanatomical atlas initiatives. We further demonstrate (for the first time the feasibility and quality of ultra-high spatial resolution (150 µm isotopic imaging of the entire human brain MRI at 7T, offering new opportunities for analyses on MR-derived information.

  13. Histological observation and occurrence of apoptosis in the gill from estuary Amazon fish

    Directory of Open Access Journals (Sweden)

    Zélia Maria Nunes

    2015-11-01

    Full Text Available Sciades herszbergii is a fish species commomly know as "blue catfish" and is one the most important natural resources an estuary in the Amazon region. In this study histological observations and apoptosis analysis was made in a gill tissue of the S. herszbergii from two environments: I, away from pollution sources and; II, waste from industry, the last one was used to relate the health status of the fishes in the environmental. Gill fragments were fixed 4% paraformaldehyde solution for 24 h and submitted for histological routine processing for embedding in paraffin. Sections of 7 μm were stained with Hematoxylin-Eosin for analysis under light microscopy. For immunohistochemycal analyses gill sections were incubated in the primary polyclonal antibody anti-caspase 3, made in rabbit. Slides were revealed using 3,3'-diaminobenzidine. The results showed the type and level of lesions were verified between two study sites. A variety of severe damages were observed in specimens including aneurysms, epithelial lifting, and intense hyperplasia, specially on the site II. Apoptosis analyses also revealed injuries in the gill tissue of specimens from these two sites. While on site I only few specimens showed mild changes in gill tissues and rare apoptosis marking. These results suggest the gill histological and immunohistochemical analysis are a good biomarker, and the specie S. herszbergii could be used as a bioindicator for environmental monitoring.

  14. Visualization of superparamagnetic nanoparticles in vascular tissue using XμCT and histology.

    Science.gov (United States)

    Tietze, Rainer; Rahn, Helene; Lyer, Stefan; Schreiber, Eveline; Mann, Jenny; Odenbach, Stefan; Alexiou, Christoph

    2011-02-01

    In order to increase the dose of antineoplastic agents in the tumor area, the concept of magnetic drug targeting (MDT) has been developed. Magnetic nanoparticles consisting of iron oxide and a biocompatible cover layer suspended in an aqueous solution (ferrofluid) serve as carriers for chemotherapeutics being enriched by an external magnetic field after intra-arterial application in desired body compartments (i.e., tumor). We established an ex vivo model to simulate in vivo conditions in a circulating system consisting of magnetic iron oxide nanoparticles passing an intact bovine artery and being focused by an external magnetic field to study their distribution in the vessel. Micro-computed X-ray tomography (XμCT) and histology can elucidate the arrangement of these particles after application. XμCT-analysis has been performed on arterial sections after MDT in order to determine the distribution of the nanoparticles. These measurements have been carried out with a cone X-ray source and corresponding histological sections were stained with Prussian blue. It could be shown that combining XμCT and histology offers the opportunity for a better understanding of the mechanisms of nanoparticle deposition in the vascular system after MDT.

  15. Clinical and histological evaluation of large macular hole surgery using the inverted internal limiting membrane flap technique

    Directory of Open Access Journals (Sweden)

    Kase S

    2016-12-01

    Full Text Available Satoru Kase, Wataru Saito, Shohei Mori, Michiyuki Saito, Ryo Ando, Zhenyu Dong, Tomohiro Suzuki, Kousuke Noda, Susumu Ishida Department of Ophthalmology, Hokkaido University Graduate School of Medicine, Sapporo, Japan Purpose: The aims of this study were to analyze optical coherence tomography (OCT imaging of large macular holes (MHs treated with inverted internal limiting membrane (ILM flap technique and to perform a histological examination of an ILM-like membrane tissue obtained during vitrectomy.Patients and methods: This is a retrospective observational case study. Nine patients, comprising of five males and four females, showing large and myopic MHs, underwent pars plana vitrectomy (PPV with inverted ILM flap technique assisted by brilliant blue G (BBG staining. Ophthalmological findings including visual acuity and OCT were investigated based on medical records. Formalin-fixed paraffin-embedded tissue section of an ILM-like membrane was submitted for immunohistochemistry with glial fibrillary acidic protein (GFAP.Results: ILM was clearly stained with BBG in eight patients, whereas the ILM in one case revealed no staining with BBG during PPV. Visual acuities improved to >0.2 LogMAR in six patients. The complete closure of MH following PPV with inverted ILM technique was eventually achieved in all patients determined by OCT imaging (100%. Only one patient showed recovery of ellipsoid zone and interdigitation zone following the surgery. Elongation of outer nuclear layer was noted in three eyes. The ILM-like membrane not stained with BBG histologically revealed an amorphous structure admixed with GFAP-positive mononuclear cell infiltration.Conclusion: PPV with inverted ILM flap technique achieved 100% closure rates with favorable configuration at an initial surgery in large MHs. Our histopathological data also suggest that even BBG staining-negative membrane may be a useful material for autologous transplantation to the hole. Keywords

  16. Development of the rat larynx: a histological study.

    Science.gov (United States)

    Alli, Opeyemi; Berzofsky, Craig; Sharma, Sansar; Pitman, Michael J

    2013-12-01

    To evaluate and describe the cartilaginous and muscular development of the rat larynx. Histologic evaluation. The larynges of Sprague Dawley rats of embryonic day (E) 13, 15, 17, 19, 21, postnatal day 0, 14, and adult of 250 gm were collected. Four larynges of each age were harvested, cut into 15-μm serial sections, stained with hematoxylin and eosin, and evaluated under light microscopy. Representative digital images were recorded and evaluated at the preglottic (supraglottic in humans), glottic, and postglottic (subglottic in humans) levels. Brachial arches were observed at E13. At E17, immature structures of the larynx, including skeletal muscle, cartilage, and the lumen were identifiable. Chondrification and muscle formation were clearly seen by E19. The muscular and cartilagenous components of the larynx were well established by E21. During the span between birth and adult maturation, the size of the larynx increased from a height of 1.10 mm to 2.90 mm, and from a width of 1.80 mm to 5.40 mm, and from a length of 1.38 mm to 4.77 mm in the stained section. Although developed at E21, the laryngeal structures continued to grow by approximately 30%. Rat laryngeal development parallels that in mice and humans. In the rat, at E17 immature structures of the larynx are identifiable, they are well developed at birth and grow by approximately 30% into adulthood. Understanding the chronology and morphology of the embryogenesis of the rat laryngeal musculature is essential and will allow for further evaluation of the embryologic innervation of these muscles. Copyright © 2013 The American Laryngological, Rhinological and Otological Society, Inc.

  17. Differentiation of meningiomas from histologic mimics via the use of claudin-1

    International Nuclear Information System (INIS)

    Shalaby, A.M.R.; Naquib, S.M.

    2006-01-01

    Meningiomas may be occasionally difficult to distinguish pathologically from other tumors of the central nervous system. Claudin-1 is a tight junction-associated protein recently shown to be expressed in anaplastic meningiomas. This study aimed at assessment whether immunohistochemical staining for claudin-1 could help distinguish meningiomas from histologic mimics, compared with commonly used markers. Tissue sections from 20 meningothelial meningiomas, 40 fibrous meningiomas, 20 atypical meningiomas, 14 solitary fibrous tumors of the meninges, 10 meningeal hemangiopericytomas, and 14 vestibular schwannomas were stained immunohistochemically for claudin-1, epithelial membrane antigen, S-100 protein, CD34, and glial fibrillary acidic protein. In total, 42(53%) of 80 meningiomas were immunoreactive for claudin-1, whereas none of the other tumors were positive. In contrast, there was considerable overlap in the distribution of the other antibodies evaluated. Claudin-1 seems to be a specific marker for meningiomas in this context. Although its sensitivity is relatively low, claudin-1 may be helpful in a panel of immunostains to distinguish meningiomas from histologic mimics. (author)

  18. Gram staining with an automatic machine.

    Science.gov (United States)

    Felek, S; Arslan, A

    1999-01-01

    This study was undertaken to develop a new Gram-staining machine controlled by a micro-controller and to investigate the quality of slides that were stained in the machine. The machine was designed and produced by the authors. It uses standard 220 V AC. Staining, washing, and drying periods are controlled by a timer built in the micro-controller. A software was made that contains a certain algorithm and time intervals for the staining mode. One-hundred and forty smears were prepared from Escherichia coli, Staphylococcus aureus, Neisseria sp., blood culture, trypticase soy broth, direct pus and sputum smears for comparison studies. Half of the slides in each group were stained with the machine, the other half by hand and then examined by four different microbiologists. Machine-stained slides had a higher clarity and less debris than the hand-stained slides (p stained slides, some Gram-positive organisms showed poor Gram-positive staining features (p Gram staining with the automatic machine increases the staining quality and helps to decrease the work load in a busy diagnostic laboratory.

  19. A generally applicable sequential alkaline phosphatase immunohistochemical double staining

    NARCIS (Netherlands)

    van der Loos, Chris M.; Teeling, Peter

    2008-01-01

    A universal type of sequential double alkaline phosphatase immunohistochemical staining is described that can be used for formalin-fixed, paraffin-embedded and cryostat tissue sections from human and mouse origin. It consists of two alkaline phosphatase detection systems including enzymatic

  20. The use of special stains in liver biopsy interpretation: Implications ...

    African Journals Online (AJOL)

    2015-12-03

    Dec 3, 2015 ... Key words: Biliary disease, iron overload, liver biopsy, special stains. Date of Acceptance: 03-Dec- .... effect of cutting fresh sections after trimming. This limits ... alcohol as a significant factor in the iron deposition found in these ...

  1. A comparision of modified and standard papanicolaou staining ...

    African Journals Online (AJOL)

    Objective: To compare modified and standard Papanicolaou (Pap) staining methods in the assessment of the cervical smears. Design: A descriptive cross sectional study. setting: Kenyatta National Hospital. Subjects: One hundred and sixty two women who were eligible for a pap smear and met the inclusion criteria.

  2. Validity of gradient-echo three-dimensional delayed gadolinium-enhanced magnetic resonance imaging of hip joint cartilage: A histologically controlled study

    Energy Technology Data Exchange (ETDEWEB)

    Zilkens, Christoph, E-mail: christoph.zilkens@med.uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Orthopaedic Surgery, Moorenstraße 5, D-40225 Dusseldorf (Germany); Miese, Falk, E-mail: falk.miese@med.uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Diagnostic and Interventional Radiology, Moorenstraße 5, D-40225 Dusseldorf (Germany); Herten, Monika, E-mail: Moherten@web.de [Univ Dusseldorf, Medical Faculty, Department of Orthopaedic Surgery, Moorenstraße 5, D-40225 Dusseldorf (Germany); Kurzidem, Sabine, E-mail: sabine.kurzidem@uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Orthopaedic Surgery, Moorenstraße 5, D-40225 Dusseldorf (Germany); Jäger, Marcus [Univ Essen, Medical Faculty, Department of Orthopaedic Surgery, D-45147 Essen (Germany); König, Dietmar, E-mail: Dietmarpierre.koenig@lvr.de [LVR Clinic for Orthopedic Surgery, D-41749 Viersen (Germany); Antoch, Gerald, E-mail: antoch@med.uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Diagnostic and Interventional Radiology, Moorenstraße 5, D-40225 Dusseldorf (Germany); Krauspe, Rüdiger, E-mail: krauspe@med.uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Orthopaedic Surgery, Moorenstraße 5, D-40225 Dusseldorf (Germany); Bittersohl, Bernd, E-mail: bernd.bittersohl@med.uni-duesseldorf.de [Univ Dusseldorf, Medical Faculty, Department of Orthopaedic Surgery, Moorenstraße 5, D-40225 Dusseldorf (Germany)

    2013-02-15

    Objective: To validate gradient-echo three-dimensional (3D) delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) by means of histological analyses in the assessment of hip joint cartilage. Materials and methods: Twenty-one femoral head specimens collected from 21 patients (7 males, 14 females, mean age: 60.9 ± 9.6 years; range: 37.6–77.3 years), who underwent total hip replacement for symptomatic hip joint osteoarthritis, underwent MRI and histological assessment. A region of 2 cm{sup 2} at the weight-bearing area was marked with four pins to enable multi-planar MRI reformatting to be matched with histological sections. MRI was performed at 3 T with a 3D double-echo steady-state (DESS) sequence for morphological cartilage assessment and 3D Volumetric Interpolated Breathhold Examination (VIBE) for T1{sub Gd} mapping. Histological sections were evaluated according to the Mankin score system. Total Mankin score, grade of toluidine staining (sensitive for glycosaminoglycan content) and a modified Mankin score classification system with four sub-groups of cartilage damage were correlated with MRI data. Results: Spearman's rho correlation analyses revealed a statistically significant correlation between T1{sub Gd} mapping and histological analyses in all categories including total Mankin score (r = −0.658, p-value ≤ 0.001), toluidine staining (r = −0.802, p-value < 0.001) and modified Mankin score (r = −0.716, p-value < 0.001). The correlation between morphological MRI and histological cartilage assessment was statistically significant but inferior to the biochemical cartilage MRI (r-values ranging from −0.411 to 0.525, p-values < 0.001). Conclusions: Gradient-echo dGEMRIC is reliable while offering the unique features of high image resolution and 3D biochemically sensitive MRI for the assessment of early cartilage degeneration.

  3. Histologically Measured Cardiomyocyte Hypertrophy Correlates with Body Height as Strongly as with Body Mass Index

    Directory of Open Access Journals (Sweden)

    Richard E. Tracy

    2011-01-01

    Full Text Available Cardiac myocytes are presumed to enlarge with left ventricular hypertrophy (LVH. This study correlates histologically measured myocytes with lean and fat body mass. Cases of LVH without coronary heart disease and normal controls came from forensic autopsies. The cross-sectional widths of myocytes in H&E-stained paraffin sections followed log normal distributions almost to perfection in all 104 specimens, with constant coefficient of variation across the full range of ventricular weight, as expected if myocytes of all sizes contribute proportionately to hypertrophy. Myocyte sizes increased with height. By regression analysis, height2.7 as a proxy for lean body mass and body mass index (BMI as a proxy for fat body mass, exerted equal effects in the multiple correlation with myocyte volume, and the equation rejected race and sex. In summary, myocyte sizes, as indexes of LVH, suggest that lean and fat body mass may contribute equally.

  4. Large-scale computations on histology images reveal grade-differentiating parameters for breast cancer

    Directory of Open Access Journals (Sweden)

    Katsinis Constantine

    2006-10-01

    Full Text Available Abstract Background Tumor classification is inexact and largely dependent on the qualitative pathological examination of the images of the tumor tissue slides. In this study, our aim was to develop an automated computational method to classify Hematoxylin and Eosin (H&E stained tissue sections based on cancer tissue texture features. Methods Image processing of histology slide images was used to detect and identify adipose tissue, extracellular matrix, morphologically distinct cell nuclei types, and the tubular architecture. The texture parameters derived from image analysis were then applied to classify images in a supervised classification scheme using histologic grade of a testing set as guidance. Results The histologic grade assigned by pathologists to invasive breast carcinoma images strongly correlated with both the presence and extent of cell nuclei with dispersed chromatin and the architecture, specifically the extent of presence of tubular cross sections. The two parameters that differentiated tumor grade found in this study were (1 the number density of cell nuclei with dispersed chromatin and (2 the number density of tubular cross sections identified through image processing as white blobs that were surrounded by a continuous string of cell nuclei. Classification based on subdivisions of a whole slide image containing a high concentration of cancer cell nuclei consistently agreed with the grade classification of the entire slide. Conclusion The automated image analysis and classification presented in this study demonstrate the feasibility of developing clinically relevant classification of histology images based on micro- texture. This method provides pathologists an invaluable quantitative tool for evaluation of the components of the Nottingham system for breast tumor grading and avoid intra-observer variability thus increasing the consistency of the decision-making process.

  5. 3D morphological measurement of whole slide histological vasculature reconstructions

    Science.gov (United States)

    Xu, Yiwen; Pickering, J. G.; Nong, Zengxuan; Ward, Aaron D.

    2016-03-01

    Properties of the microvasculature that contribute to tissue perfusion can be assessed using immunohistochemistry on 2D histology sections. However, the vasculature is inherently 3D and the ability to measure and visualize the vessel wall components in 3D will aid in detecting focal pathologies. Our objectives were (1) to develop a method for 3D measurement and visualization of microvasculature in 3D, (2) to compare the normal and regenerated post-ischemia mouse hind limb microvasculature, and (3) to compare the 2D and 3D vessel morphology measures. Vessels were stained for smooth muscle using 3,3'-Diaminobenzidine (DAB) immunostain for both normal (n = 6 mice) and regenerated vasculature (n = 5 mice). 2D vessel segmentations were reconstructed into 3D using landmark based registration. No substantial bias was found in the 2D measurements relative to 3D, but larger differences were observed for individual vessels oriented non-orthogonally to the plane of sectioning. A larger value of area, perimeter, and vessel wall thickness was found in the normal vasculature as compared to the regenerated vasculature, for both the 2D and 3D measurements (p pathologies on a whole slide level.

  6. On the anatomy and histology of the pubovisceral muscle enthesis in women.

    Science.gov (United States)

    Kim, Jinyong; Ramanah, Rajeev; DeLancey, John O L; Ashton-Miller, James A

    2011-09-01

    The origin of the pubovisceral muscle (PVM) from the pubic bone is known to be at elevated risk for injury during difficult vaginal births. We examined the anatomy and histology of its enthesial origin to classify its type and see if it differs from appendicular entheses. Parasagittal sections of the pubic bone, PVM enthesis, myotendinous junction, and muscle proper were harvested from five female cadavers (51-98 years). Histological sections were prepared with hematoxylin and eosin, Masson's trichrome, and Verhoeff-Van Gieson stains. The type of enthesis was identified according to a published enthesial classification scheme. Quantitative imaging analysis was performed in sampling bands 2 mm apart along the enthesis to determine its cross-sectional area and composition. The PVM enthesis can be classified as a fibrous enthesis. The PVM muscle fibers terminated in collagenous fibers that insert tangentially onto the periosteum of the pubic bone for the most part. Sharpey's fibers were not observed. In a longitudinal cross-section, the area of the connective tissue and muscle becomes equal approximately 8 mm from the pubic bone. The PVM originates bilaterally from the pubic bone via fibrous entheses whose collagen fibers arise tangentially from the periosteum of the pubic bone. Copyright © 2010 Wiley-Liss, Inc.

  7. Peripheral ovine progressive pneumonia provirus levels correlate with and predict histological tissue lesion severity in naturally infected sheep.

    Science.gov (United States)

    Herrmann-Hoesing, Lynn M; Noh, Susan M; White, Stephen N; Snekvik, Kevin R; Truscott, Thomas; Knowles, Donald P

    2009-04-01

    Studies were undertaken to determine whether anti-ovine progressive pneumonia virus (OPPV) antibody responses in serum or OPP provirus levels in peripheral blood associate with the degree of histologically measured tissue lesions in naturally OPPV-infected sheep. Sections of formalin-fixed, paraffin-embedded, and hematoxylin- and eosin-stained lung, mammary gland, carpal synovial membrane, and brain tissues from 11 OPPV-infected ewes (mean age of 8.6 years) and 5 OPPV-uninfected ewes (mean age of 6 years) were evaluated for lesion severity. Ovine progressive pneumonia (OPP) provirus levels and anti-OPPV antibody titers in peripheral blood and serum samples, respectively, were measured upon euthanasia and 3 years prior to euthanasia. Both mean peripheral OPP provirus levels and mean serum anti-surface envelope glycoprotein (anti-SU) antibody titers at the time of euthanasia were significantly higher in ewes with moderate to severe histological lesions than in ewes with no to mild histological lesions. However, although mean peripheral blood OPP provirus levels at euthanasia and 3 years prior to euthanasia significantly correlated with the highest histological lesion score for any affected tissue (two-tailed P values, 0.03 and 0.02), mean serum anti-SU antibody titers, anti-capsid antibody titers, and anti-transmembrane 90 antibody titers at euthanasia did not show a significant correlation with the highest histological lesion score for any tissue (two-tailed P values, 0.32, 0.97, and 0.18, respectively). These data are the first to show that OPP provirus levels predict and correlate with the extent of OPPV-related histological lesions in various OPPV-affected tissues. These findings suggest that peripheral OPP provirus levels quantitatively contribute more to the development of histological lesions than the systemic anti-SU antibody host immune response.

  8. Gastritis: the histology report.

    Science.gov (United States)

    Rugge, Massimo; Pennelli, Gianmaria; Pilozzi, Emanuela; Fassan, Matteo; Ingravallo, Giuseppe; Russo, Valentina M; Di Mario, Francesco

    2011-03-01

    Gastritis is defined as inflammation of the gastric mucosa. In histological terms, it is distinguishable into two main categories, i.e. non-atrophic and atrophic. In the gastric mucosa, atrophy is defined as the loss of appropriate glands. There are several etiological types of gastritis, their different etiology being related to different clinical manifestations and pathological features. Atrophic gastritis (resulting mainly from long-standing Helicobacter pylori infection) is a major risk factor for the onset of (intestinal type) gastric cancer. The extent and site of the atrophic changes correlate significantly with the cancer risk. The current format for histology reporting in cases of gastritis fails to establish an immediate link between gastritis phenotype and risk of malignancy. Building on current knowledge of the biology of gastritis, an international group of pathologists [Operative Link for Gastritis Assessment (OLGA)] has proposed a system for reporting gastritis in terms of its stage (the OLGA Staging System): this system places the histological phenotypes of gastritis on a scale of progressively increasing gastric cancer risk, from the lowest (Stage 0) to the highest (Stage IV). The aim of this tutorial is to provide unequivocal information on how to standardize histology reports on gastritis in diagnostic practice. Copyright © 2011 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd.. All rights reserved.

  9. Anatomical analysis of an aye-aye brain (Daubentonia madagascariensis, primates: Prosimii) combining histology, structural magnetic resonance imaging, and diffusion-tensor imaging.

    Science.gov (United States)

    Kaufman, Jason A; Ahrens, Eric T; Laidlaw, David H; Zhang, Song; Allman, John M

    2005-11-01

    This report presents initial results of a multimodal analysis of tissue volume and microstructure in the brain of an aye-aye (Daubentonia madagascariensis). The left hemisphere of an aye-aye brain was scanned using T2-weighted structural magnetic resonance imaging (MRI) and diffusion-tensor imaging (DTI) prior to histological processing and staining for Nissl substance and myelinated fibers. The objectives of the experiment were to estimate the volume of gross brain regions for comparison with published data on other prosimians and to validate DTI data on fiber anisotropy with histological measurements of fiber spread. Measurements of brain structure volumes in the specimen are consistent with those reported in the literature: the aye-aye has a very large brain for its body size, a reduced volume of visual structures (V1 and LGN), and an increased volume of the olfactory lobe. This trade-off between visual and olfactory reliance is likely a reflection of the nocturnal extractive foraging behavior practiced by Daubentonia. Additionally, frontal cortex volume is large in the aye-aye, a feature that may also be related to its complex foraging behavior and sensorimotor demands. Analysis of DTI data in the anterior cingulum bundle demonstrates a strong correlation between fiber spread as measured from histological sections and fiber spread as measured from DTI. These results represent the first quantitative comparison of DTI data and fiber-stained histology in the brain. (c) 2005 Wiley-Liss, Inc.

  10. Magnetic Resonance Imaging of Japanese monkey brains compared with X-ray photography and histology

    International Nuclear Information System (INIS)

    Kaji, Shinji; Matsuda, Keiji; Kawano, Kenji; Komatsu, Hidehiko; Yamane, Shigeru; Yoshizawa, Takashi; Nose, Tadao.

    1991-01-01

    The localization of a small target area in the brain is usually estimated by using stereotaxic atlases, assisted by X-ray photography or electrophysiological mapping, and determined finally by histological reconstruction. Magnetic Resonance Imaging (MRI) can visualize, noninvasively cross sections in any plane of three dimensional structures of the brain. We compared images of MRI, X-ray, and histology from a monkey brain. A Japanese monkey (Macaca fuscata, male, 11 kg) was anesthetised, fixed in a newly developed magnetic-free stereotaxic apparatus, and mounted in the MRI scanner unit (BRUKER, BIOSPEC 24/40, 2.4 Tesla). Some para-saggital (5 mm thick) and para-frontal (2.5 mm thick, every 5 mm distance) images were obtained. The outline of the bone on the MRI image was compared with that on the X-ray photograph taken by an X-ray instrument (Toshiba, TR-80A). The two images fitted very well. The animal was sacrificed, the brain was sliced in 100 μm and stained with Cresyl violet. The histological preparations were shrunk some 10 % during the process, which was revealed by comparison of MRI, X-ray, and histological images. In conclusion, MRI images are reliable enough to determine a small target in deep structures of the brain, and their superimposed images on X-rays will assist in identifying the location of electrode or needle tips. We constructed a data base of these MRI and histological images on a Macintosh computer, and they can be easily accessed by a mouse operation. (author)

  11. Multi-modal image registration: matching MRI with histology

    Science.gov (United States)

    Alic, Lejla; Haeck, Joost C.; Klein, Stefan; Bol, Karin; van Tiel, Sandra T.; Wielopolski, Piotr A.; Bijster, Magda; Niessen, Wiro J.; Bernsen, Monique; Veenland, Jifke F.; de Jong, Marion

    2010-03-01

    Spatial correspondence between histology and multi sequence MRI can provide information about the capabilities of non-invasive imaging to characterize cancerous tissue. However, shrinkage and deformation occurring during the excision of the tumor and the histological processing complicate the co registration of MR images with histological sections. This work proposes a methodology to establish a detailed 3D relation between histology sections and in vivo MRI tumor data. The key features of the methodology are a very dense histological sampling (up to 100 histology slices per tumor), mutual information based non-rigid B-spline registration, the utilization of the whole 3D data sets, and the exploitation of an intermediate ex vivo MRI. In this proof of concept paper, the methodology was applied to one tumor. We found that, after registration, the visual alignment of tumor borders and internal structures was fairly accurate. Utilizing the intermediate ex vivo MRI, it was possible to account for changes caused by the excision of the tumor: we observed a tumor expansion of 20%. Also the effects of fixation, dehydration and histological sectioning could be determined: 26% shrinkage of the tumor was found. The annotation of viable tissue, performed in histology and transformed to the in vivo MRI, matched clearly with high intensity regions in MRI. With this methodology, histological annotation can be directly related to the corresponding in vivo MRI. This is a vital step for the evaluation of the feasibility of multi-spectral MRI to depict histological groundtruth.

  12. Liquid dish washing soap: An excellent substitute for xylene and alcohol in hematoxylin and eosin staining procedure

    Directory of Open Access Journals (Sweden)

    Surekha Ramulu

    2012-01-01

    Full Text Available Aims: Liquid dish washing solution (DWS was used as a substitute for xylene to dewax tissue sections during hematoxylin and eosin (H and E staining. The aim was to test and compare the hypothesis that xylene-ethanol free (XEF sections deparaffinized with diluted DWS are better than or at par with the conventional H and E sections. Materials and Methods: Fifty paraffin-embedded tissue blocks was included. One section was stained with conventional HandE (group A and the other with XEF HandE (group B staining method. Slides were scored for parameters: nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. Z test was used for statistical analysis. For accuracy of diagnosis, sensitivity, specificity, positive predictive value, and negative predictive value were tested. Results: Adequate nuclear staining was noted in 94% in group A and 96% in group B, -adequate cytoplasmic staining in 92% in group A and 86% in group B, clarity in 94% of group A and 96% of group B sections, uniform staining in 92% of group A and 80% of group B sections, crisp stain in 96% of group A and 88% of group B sections, and 94% of group A sections stained adequately for diagnosis as compared with 90% in group B sections. Conclusion: Liquid DWS can be used as an alternative and effective substitute to xylene and ethanol in routine HandE staining procedure.

  13. Comparison of staining of mitotic figures by haematoxylin and eosin-and crystal violet stains, in oral epithelial dysplasia and squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Ankle Madhuri

    2007-01-01

    Full Text Available Mitosis of cells gives rise to tissue integrity. Defects during mitosis bring about abnormalities. Excessive proliferation of cells due to increased mitosis is one such outcome, which is the hallmark in precancer and cancer. The localization of proliferating cells or their precursors may not be obvious and easy. Establishing an easy way to distinguish these mitotic cells will help in grading and understanding their biological potential. Although immunohistochemistry is an advanced method in use, the cost and time factor makes it less feasible for many laboratories. Selective histochemical stains like toluidine blue, giemsa and crystal violet have been used in tissues including the developing brain, neural tissue and skin. Aim of the study: 1To compare the staining of mitotic cells in haematoxylin and eosin with that in crystal violet. 2To compare the number of mitotic figures present in normal oral mucosa, epithelial dysplasia and oral squamous cell carcinoma in crystal violet-stained sections with that in H and E-stained sections. Materials and Methods: Ten tissues of normal oral mucosa and 15 tissues each of oral epithelial dysplasia seen in tobacco-associated leukoplakia and squamous cell carcinoma were studied to evaluate the selectivity of 1% crystal violet for mitotic figures. The staining was compared with standard H and E staining. Statistical analysis was done using Man-Whitney U test. Results: A statistically significant increase in the mean mitotic count was observed in crystal violet-stained sections of epithelial dysplasia as compared to the H and E-stained sections ( p = 0.0327. A similar increase in the mitotic counts was noted in crystal violet-stained sections of oral squamous cell carcinoma as compared to the H and E-stained sections.( p = 0.0443. No significant difference was found in the mitotic counts determined in dysplasia or carcinoma by either the crystal violet ( p = 0.4429 or the H and E-staining techniques ( p = 0

  14. One Size Fits All: Evaluation of the Transferability of a New "Learning" Histologic Image Analysis Application.

    Science.gov (United States)

    Arlt, Janine; Homeyer, André; Sänger, Constanze; Dahmen, Uta; Dirsch, Olaf

    2016-01-01

    Quantitative analysis of histologic slides is of importance for pathology and also to address surgical questions. Recently, a novel application was developed for the automated quantification of whole-slide images. The aim of this study was to test and validate the underlying image analysis algorithm with respect to user friendliness, accuracy, and transferability to different histologic scenarios. The algorithm splits the images into tiles of a predetermined size and identifies the tissue class of each tile. In the training procedure, the user specifies example tiles of the different tissue classes. In the subsequent analysis procedure, the algorithm classifies each tile into the previously specified classes. User friendliness was evaluated by recording training time and testing reproducibility of the training procedure of users with different background. Accuracy was determined with respect to single and batch analysis. Transferability was demonstrated by analyzing tissue of different organs (rat liver, kidney, small bowel, and spleen) and with different stainings (glutamine synthetase and hematoxylin-eosin). Users of different educational background could apply the program efficiently after a short introduction. When analyzing images with similar properties, accuracy of >90% was reached in single images as well as in batch mode. We demonstrated that the novel application is user friendly and very accurate. With the "training" procedure the application can be adapted to novel image characteristics simply by giving examples of relevant tissue structures. Therefore, it is suitable for the fast and efficient analysis of high numbers of fully digitalized histologic sections, potentially allowing "high-throughput" quantitative "histomic" analysis.

  15. Serum Matrix Metalloproteinase-3 as a Noninvasive Biomarker of Histological Synovitis for Diagnosis of Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Jian-Da Ma

    2014-01-01

    Full Text Available Objective. To explore the correlation between matrix metalloproteinase- (MMP- 3 and histological synovitis in rheumatoid arthritis (RA. Methods. Serum MMP-3 of 62 patients with active RA was detected by ELISA. Serial synovial tissue sections from all RA patients, 13 osteoarthritis, and 10 orthopedic arthropathies patients were stained with hematoxylin and eosin and immunohistochemically for MMP-3, CD3, CD20, CD38, CD68, and CD15. Results. The percentage of lining MMP3+ cells was significantly higher in RA patients especially with high grade synovitis and it was significantly correlated with Krenn’s synovitis score r=0.574, P<0.001 and sublining inflammatory cells. Multivariate stepwise linear regression analysis revealed that the association of the percentage of lining MMP3+ cells with activation of synovial stroma, sublining CD68+ macrophages, and CD15+ neutrophils was stronger than other histological indicators. The percentage of lining MMP3+ cells was significantly correlated with serum MMP-3 in RA r=0.656, P<0.001. Serum MMP-3 was higher in RA patients with high grade synovitis than that of low grade synovitis and significantly correlated with synovitis score and activation of synovial stroma subscore (all P<0.05. Conclusion. Serum MMP-3 may be an alternative noninvasive biomarker of histological synovitis and RA diagnosis.

  16. Hirschsprung′s disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining

    Directory of Open Access Journals (Sweden)

    Memarzadeh Mehrdad

    2009-01-01

    Full Text Available Background : The diagnosis of Hirschsprung′s disease (HD is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods : In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staining slides were compared with immunohistochemical (IHC staining including: S100, NSE, CD117, CD56, Cathepsin D, Vimentin, BCL2, GFAP, Synaptophysin and chromogranin. Results : The study revealed 30 negative (absence of ganglion cells cases (55.5%, 17 positive cases (31.04% and seven suspected cases (12.9% of ganglion cells on the H and E staining. On IHC staining with CD56 and Cathepsin D, all of the 17 positive cases detected through H and E, were confirmed for having ganglion cells and out of 30 cases reported negative on H and E staining, 28(93.3% were reported negative and two (6.7% positive by IHC staining. Of the seven suspected cases H and E staining, IHC staining detectedganglion cells only in five slides; two remained negative. Conclusions : IHC staining using CD56 and Cathepsin D improved the accuracy of diagnosis in HD when used in addition to H and E staining technique, especially for negative or suspicious slides.

  17. SMASH - semi-automatic muscle analysis using segmentation of histology: a MATLAB application.

    Science.gov (United States)

    Smith, Lucas R; Barton, Elisabeth R

    2014-01-01

    Histological assessment of skeletal muscle tissue is commonly applied to many areas of skeletal muscle physiological research. Histological parameters including fiber distribution, fiber type, centrally nucleated fibers, and capillary density are all frequently quantified measures of skeletal muscle. These parameters reflect functional properties of muscle and undergo adaptation in many muscle diseases and injuries. While standard operating procedures have been developed to guide analysis of many of these parameters, the software to freely, efficiently, and consistently analyze them is not readily available. In order to provide this service to the muscle research community we developed an open source MATLAB script to analyze immunofluorescent muscle sections incorporating user controls for muscle histological analysis. The software consists of multiple functions designed to provide tools for the analysis selected. Initial segmentation and fiber filter functions segment the image and remove non-fiber elements based on user-defined parameters to create a fiber mask. Establishing parameters set by the user, the software outputs data on fiber size and type, centrally nucleated fibers, and other structures. These functions were evaluated on stained soleus muscle sections from 1-year-old wild-type and mdx mice, a model of Duchenne muscular dystrophy. In accordance with previously published data, fiber size was not different between groups, but mdx muscles had much higher fiber size variability. The mdx muscle had a significantly greater proportion of type I fibers, but type I fibers did not change in size relative to type II fibers. Centrally nucleated fibers were highly prevalent in mdx muscle and were significantly larger than peripherally nucleated fibers. The MATLAB code described and provided along with this manuscript is designed for image processing of skeletal muscle immunofluorescent histological sections. The program allows for semi-automated fiber detection

  18. Comparison of different histological protocols for the preservation and quantification of the intestinal mucus layer in pigs

    Directory of Open Access Journals (Sweden)

    Ilen Röhe

    2018-02-01

    Full Text Available The histological characterization of the intestinal mucus layer is important for many scientific experiments investigating the interaction between intestinal microbiota, mucosal immune response and intestinal mucus production. The aim of this study was to examine and compare different fixation protocols for displaying and quantifying the intestinal mucus layer in piglets and to test which histomorphological parameters may correlate with the determined mucus layer thickness. Jejunal and colonal tissue samples of weaned piglets (n=10 were either frozen in liquid nitrogen or chemically fixed using methacarn solution. The frozen tissue samples were cryosectioned and subsequently postfixed using three different postfixatives: paraformaldehyde vapor, neutrally buffered formalin solution and ethanol solution. After dehydration, methacarn fixed tissues were embedded in paraffin wax. Both sections of cryopreserved and methacarn fixed tissue samples were stained with Alcian blue (AB-PAS followed by the microscopically determination of the mucus layer thickness. Different pH values of the Alcian Blue staining solution and two mucus layer thickness measuring methods were compared. In addition, various histomorphological parameters of methacarn fixed tissue samples were evaluated including the number of goblet cells and the mucin staining area. Cryopreservation in combination with chemical postfixation led to mucus preservation in the colon of piglets allowing mucus thickness measurements. Mucus could be only partly preserved in cryosections of the jejunum impeding any quantitative description of the mucus layer thickness. The application of different postfixations, varying pH values of the AB solution and different mucus layer measuring methods led to comparable results regarding the mucus layer thickness. Methacarn fixation proved to be unsuitable for mucus depiction as only mucus patches were found in the jejunum or a detachment of the mucus layer from

  19. Comparison of different histological protocols for the preservation and quantification of the intestinal mucus layer in pigs.

    Science.gov (United States)

    Röhe, Ilen; Hüttner, Friedrich Joseph; Plendl, Johanna; Drewes, Barbara; Zentek, Jürgen

    2018-02-05

    The histological characterization of the intestinal mucus layer is important for many scientific experiments investigating the interaction between intestinal microbiota, mucosal immune response and intestinal mucus production. The aim of this study was to examine and compare different fixation protocols for displaying and quantifying the intestinal mucus layer in piglets and to test which histomorphological parameters may correlate with the determined mucus layer thickness. Jejunal and colonal tissue samples of weaned piglets (n=10) were either frozen in liquid nitrogen or chemically fixed using methacarn solution. The frozen tissue samples were cryosectioned and subsequently postfixed using three different postfixatives: paraformaldehyde vapor, neutrally buffered formalin solution and ethanol solution. After dehydration, methacarn fixed tissues were embedded in paraffin wax. Both sections of cryopreserved and methacarn fixed tissue samples were stained with Alcian blue (AB)-PAS followed by the microscopically determination of the mucus layer thickness. Different pH values of the Alcian Blue staining solution and two mucus layer thickness measuring methods were compared. In addition, various histomorphological parameters of methacarn fixed tissue samples were evaluated including the number of goblet cells and the mucin staining area. Cryopreservation in combination with chemical postfixation led to mucus preservation in the colon of piglets allowing mucus thickness measurements. Mucus could be only partly preserved in cryosections of the jejunum impeding any quantitative description of the mucus layer thickness. The application of different postfixations, varying pH values of the AB solution and different mucus layer measuring methods led to comparable results regarding the mucus layer thickness. Methacarn fixation proved to be unsuitable for mucus depiction as only mucus patches were found in the jejunum or a detachment of the mucus layer from the epithelium was

  20. Laser treatment of Port-wine stains

    OpenAIRE

    Boffa, Michael J.

    2001-01-01

    A state-of-the-art pulsed dye laser machine to treat port-wine stains and other vascular lesions has been available in the Malta Health Service since 1999. This article reviews the pathophysiology and clinical features of port- wine stains and describes the principles of laser treatment for this condition.

  1. Histological spectrum of angiofibroma of soft tissue: histological and genetic analysis of 13 cases.

    Science.gov (United States)

    Yamada, Yuichi; Yamamoto, Hidetaka; Kohashi, Kenichi; Ishii, Takeaki; Iura, Kunio; Maekawa, Akira; Bekki, Hirofumi; Otsuka, Hiroshi; Yamashita, Kyoko; Tanaka, Hiroyuki; Hiraki, Tsubasa; Mukai, Munenori; Shirakawa, Atsuko; Shinnou, Yoko; Jinno, Mari; Yanai, Hiroyuki; Taguchi, Kenichi; Maehara, Yoshihiko; Iwamoto, Yukihide; Oda, Yosinao

    2016-09-01

    Angiofibroma of soft tissue (AFST) is a rare soft tissue neoplasm characterized by a fibroblastic cytomorphology and a prominent vascular structure. AFSTs possess a novel fusion gene, i.e. NCOA2-AHRR/AHRR-NCOA2 or GTF2I-NCOA2, providing a useful approach to diagnosing AFST. Morphologically, AFSTs span a wide spectrum, making diagnosis a challenge. The aim of this study was to review AFST cases and to report previously unknown histological features, which we confirmed by genetic analysis. We reviewed 276 cases diagnosed as solitary fibrous tumours/haemangiopericytomas (232 cases), unclassified tumours of fibroblastic differentiation (36 cases), and recently diagnosed AFSTs (eight cases), and retrieved 13 cases compatible with AFST. Immunohistochemical staining was performed for these cases, all 13 of which were analysed by reverse transcription polymerase chain reaction and fluorescence in-situ hybridization. The histological findings were as follows: amianthoid fibres, extravasation of red blood cells, haemosiderin deposition, aggregates of foamy histiocytes, cystic change, necrosis, and haemorrhage. Immunohistochemically, the tumour cells were positive for epithelial membrane antigen (four of 13 cases), desmin (six of 13 cases), CD163 (13 of 13 cases), CD68 (seven of 13 cases), oestrogen receptor (13 of 13 cases), progesterone receptor (three of 13 cases), and STAT6 (one of 13 cases, weak nuclear staining), but they were negative for CD34, α-smooth muscle actin, muscle-specific actin, S100, pan-cytokeratin, MDM2, and CDK4. The AHRR-NCOA2 fusion gene was detected in eight cases, and NCOA2 gene rearrangement in nine cases. We revealed the previously unreported histological variation and immunohistochemical findings of AFST, and confirmed them by using genetic methods. The results suggested that AFST should be considered in the diagnosis of fibrous or fibrohistiocytic tumours with the above histological features. © 2016 John Wiley & Sons Ltd.

  2. Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage

    Directory of Open Access Journals (Sweden)

    J. Rieger

    2013-07-01

    Full Text Available Staining of mast cells (MCs, including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkage-differences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from

  3. International Cartilage Repair Society (ICRS) Recommended Guidelines for Histological Endpoints for Cartilage Repair Studies in Animal Models and Clinical Trials

    Science.gov (United States)

    Hoemann, Caroline; Kandel, Rita; Roberts, Sally; Saris, Daniel B.F.; Creemers, Laura; Mainil-Varlet, Pierre; Méthot, Stephane; Hollander, Anthony P.; Buschmann, Michael D.

    2011-01-01

    Cartilage repair strategies aim to resurface a lesion with osteochondral tissue resembling native cartilage, but a variety of repair tissues are usually observed. Histology is an important structural outcome that could serve as an interim measure of efficacy in randomized controlled clinical studies. The purpose of this article is to propose guidelines for standardized histoprocessing and unbiased evaluation of animal tissues and human biopsies. Methods were compiled from a literature review, and illustrative data were added. In animal models, treatments are usually administered to acute defects created in healthy tissues, and the entire joint can be analyzed at multiple postoperative time points. In human clinical therapy, treatments are applied to developed lesions, and biopsies are obtained, usually from a subset of patients, at a specific time point. In striving to standardize evaluation of structural endpoints in cartilage repair studies, 5 variables should be controlled: 1) location of biopsy/sample section, 2) timing of biopsy/sample recovery, 3) histoprocessing, 4) staining, and 5) blinded evaluation with a proper control group. Histological scores, quantitative histomorphometry of repair tissue thickness, percentage of tissue staining for collagens and glycosaminoglycan, polarized light microscopy for collagen fibril organization, and subchondral bone integration/structure are all relevant outcome measures that can be collected and used to assess the efficacy of novel therapeutics. Standardized histology methods could improve statistical analyses, help interpret and validate noninvasive imaging outcomes, and permit cross-comparison between studies. Currently, there are no suitable substitutes for histology in evaluating repair tissue quality and cartilaginous character. PMID:26069577

  4. Multicenter Assessment of Gram Stain Error Rates.

    Science.gov (United States)

    Samuel, Linoj P; Balada-Llasat, Joan-Miquel; Harrington, Amanda; Cavagnolo, Robert

    2016-06-01

    Gram stains remain the cornerstone of diagnostic testing in the microbiology laboratory for the guidance of empirical treatment prior to availability of culture results. Incorrectly interpreted Gram stains may adversely impact patient care, and yet there are no comprehensive studies that have evaluated the reliability of the technique and there are no established standards for performance. In this study, clinical microbiology laboratories at four major tertiary medical care centers evaluated Gram stain error rates across all nonblood specimen types by using standardized criteria. The study focused on several factors that primarily contribute to errors in the process, including poor specimen quality, smear preparation, and interpretation of the smears. The number of specimens during the evaluation period ranged from 976 to 1,864 specimens per site, and there were a total of 6,115 specimens. Gram stain results were discrepant from culture for 5% of all specimens. Fifty-eight percent of discrepant results were specimens with no organisms reported on Gram stain but significant growth on culture, while 42% of discrepant results had reported organisms on Gram stain that were not recovered in culture. Upon review of available slides, 24% (63/263) of discrepant results were due to reader error, which varied significantly based on site (9% to 45%). The Gram stain error rate also varied between sites, ranging from 0.4% to 2.7%. The data demonstrate a significant variability between laboratories in Gram stain performance and affirm the need for ongoing quality assessment by laboratories. Standardized monitoring of Gram stains is an essential quality control tool for laboratories and is necessary for the establishment of a quality benchmark across laboratories. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  5. Computer-assisted assessment of the Human Epidermal Growth Factor Receptor 2 immunohistochemical assay in imaged histologic sections using a membrane isolation algorithm and quantitative analysis of positive controls

    International Nuclear Information System (INIS)

    Hall, Bonnie H; Ianosi-Irimie, Monica; Javidian, Parisa; Chen, Wenjin; Ganesan, Shridar; Foran, David J

    2008-01-01

    Breast cancers that overexpress the human epidermal growth factor receptor 2 (HER2) are eligible for effective biologically targeted therapies, such as trastuzumab. However, accurately determining HER2 overexpression, especially in immunohistochemically equivocal cases, remains a challenge. Manual analysis of HER2 expression is dependent on the assessment of membrane staining as well as comparisons with positive controls. In spite of the strides that have been made to standardize the assessment process, intra- and inter-observer discrepancies in scoring is not uncommon. In this manuscript we describe a pathologist assisted, computer-based continuous scoring approach for increasing the precision and reproducibility of assessing imaged breast tissue specimens. Computer-assisted analysis on HER2 IHC is compared with manual scoring and fluorescence in situ hybridization results on a test set of 99 digitally imaged breast cancer cases enriched with equivocally scored (2+) cases. Image features are generated based on the staining profile of the positive control tissue and pixels delineated by a newly developed Membrane Isolation Algorithm. Evaluation of results was performed using Receiver Operator Characteristic (ROC) analysis. A computer-aided diagnostic approach has been developed using a membrane isolation algorithm and quantitative use of positive immunostaining controls. By incorporating internal positive controls into feature analysis a greater Area Under the Curve (AUC) in ROC analysis was achieved than feature analysis without positive controls. Evaluation of HER2 immunostaining that utilized membrane pixels, controls, and percent area stained showed significantly greater AUC than manual scoring, and significantly less false positive rate when used to evaluate immunohistochemically equivocal cases. It has been shown that by incorporating both a membrane isolation algorithm and analysis of known positive controls a computer-assisted diagnostic algorithm was

  6. Computer-assisted assessment of the Human Epidermal Growth Factor Receptor 2 immunohistochemical assay in imaged histologic sections using a membrane isolation algorithm and quantitative analysis of positive controls

    Directory of Open Access Journals (Sweden)

    Ianosi-Irimie Monica

    2008-06-01

    Full Text Available Abstract Background Breast cancers that overexpress the human epidermal growth factor receptor 2 (HER2 are eligible for effective biologically targeted therapies, such as trastuzumab. However, accurately determining HER2 overexpression, especially in immunohistochemically equivocal cases, remains a challenge. Manual analysis of HER2 expression is dependent on the assessment of membrane staining as well as comparisons with positive controls. In spite of the strides that have been made to standardize the assessment process, intra- and inter-observer discrepancies in scoring is not uncommon. In this manuscript we describe a pathologist assisted, computer-based continuous scoring approach for increasing the precision and reproducibility of assessing imaged breast tissue specimens. Methods Computer-assisted analysis on HER2 IHC is compared with manual scoring and fluorescence in situ hybridization results on a test set of 99 digitally imaged breast cancer cases enriched with equivocally scored (2+ cases. Image features are generated based on the staining profile of the positive control tissue and pixels delineated by a newly developed Membrane Isolation Algorithm. Evaluation of results was performed using Receiver Operator Characteristic (ROC analysis. Results A computer-aided diagnostic approach has been developed using a membrane isolation algorithm and quantitative use of positive immunostaining controls. By incorporating internal positive controls into feature analysis a greater Area Under the Curve (AUC in ROC analysis was achieved than feature analysis without positive controls. Evaluation of HER2 immunostaining that utilized membrane pixels, controls, and percent area stained showed significantly greater AUC than manual scoring, and significantly less false positive rate when used to evaluate immunohistochemically equivocal cases. Conclusion It has been shown that by incorporating both a membrane isolation algorithm and analysis of known

  7. Hirschsprung's disease diagnosis: Comparison of immunohistochemical, hematoxilin and eosin staining

    OpenAIRE

    Memarzadeh, Mehrdad; Talebi, Ardeshir; Edalaty, Masod; Hosseinpour, Mehrdad; Vahidi, Nasrin

    2009-01-01

    Background: The diagnosis of Hirschsprung's disease (HD) is based on the absence of ganglion cells. In hemotoxilin and eosin (H and E) as well as acetylcholine esterase staining there are limitations in the diagnosis of immature ganglion cells in neonates. Methods: In this prospective study, 54 biopsies taken from suspected HD patients (five mucosal specimens and 49 full thickness specimens) were studied. In the laboratory, after preparing sections of paraffin embedded tissues, H and E staini...

  8. Processing large-diameter poly(L-lactic acid) microfiber mesh/mesenchymal stromal cell constructs via resin embedding: an efficient histologic method

    International Nuclear Information System (INIS)

    D’Alessandro, Delfo; Danti, Serena; Pertici, Gianni; Moscato, Stefania; Metelli, Maria Rita; Petrini, Mario; Danti, Sabrina; Berrettini, Stefano; Nesti, Claudia

    2014-01-01

    In this study, we performed a complete histologic analysis of constructs based on large diameter ( > 100 μm) poly-L-lactic acid (PLLA) microfibers obtained via dry-wet spinning and rat Mesenchymal Stromal Cells (rMSCs) differentiated towards the osteogenic lineage, using acrylic resin embedding. In many synthetic polymer-based microfiber meshes, ex post processability of fiber/cell constructs for histologic analysis may face deterring difficulties, leading to an incomplete investigation of the potential of these scaffolds. Indeed, while polymeric nanofiber (fiber diameter = tens of nanometers)/cell constructs can usually be embedded in common histologic media and easily sectioned, preserving the material structure and the antigenic reactivity, histologic analysis of large polymeric microfiber/cell constructs in the literature is really scant. This affects microfiber scaffolds based on FDA-approved and widely used polymers such as PLLA and its copolymers. Indeed, for such constructs, especially those with fiber diameter and fiber interspace much larger than cell size, standard histologic processing is usually inefficient due to inhomogeneous hardness and lack of cohesion between the synthetic and the biological phases under sectioning. In this study, the microfiber/MSC constructs were embedded in acrylic resin and the staining/reaction procedures were calibrated to demonstrate the possibility of successfully employing histologic methods in tissue engineering studies even in such difficult cases. We histologically investigated the main osteogenic markers and extracellular matrix molecules, such as alkaline phosphatase, osteopontin, osteocalcin, TGF-β1, Runx2, Collagen type I and the presence of amorphous, fibrillar and mineralized matrix. Biochemical tests were employed to confirm our findings. This protocol permitted efficient sectioning of the treated constructs and good penetration of the histologic reagents, thus allowing distribution and expression of

  9. Morphology of the tongue of the emu (Dromaius novaehollandiae. II. Histological features

    Directory of Open Access Journals (Sweden)

    M.R. Crole

    2009-09-01

    Full Text Available Although a number of brief, fragmented descriptions have been provided on the gross morphology of the ratite tongue, very few studies have documented the histological structure of this organ. This paper presents the first definitive histological description of the emu tongue and reviews, consolidates and compares the scattered information on the histology of the ratite tongue available in the literature. Five tongues were removed from heads obtained from birds at slaughter and fixed in 10 % neutral buffered formalin. Appropriate longitudinal and transverse segments were removed, routinely processed for light microscopy, and sections examined after staining with H & E and PAS. The entire tongue (body and root is invested by a non-keratinized stratified squamous epithelium. The supporting connective tissue of the tongue dorsum displays only large, simple branched tubular mucussecreting glands, whereas the caudal tongue body ventrum and tongue root, in addition to these glands, also exhibits small, simple tubular mucus-secreting glands. Herbst corpuscles are associated with the large, simple branched glands. Lymphoid tissue is restricted to the tongue ventrum and is particularly obvious at the junction of the ventral tongue body and frenulum where a large aggregation of diffuse lymphoid tissue, with nodular tissue proximally, was consistently observed. A structure resembling a taste bud was located in the epithelium on the caudal extremity of the tongue root of one bird. This is the first reported observation of taste buds in ratites. Forming the core of the tongue body is the cartilaginous paraglossum lying dorsal to the partially ossified rostral projection of the basihyale. The histological features of the emu tongue are generally similar to those described for the greater rhea and ostrich, except that taste buds were not identified in these species. The results would suggest that the emu tongue functions as a sensory organ, both for taste and

  10. Combined histochemical staining, RNA amplification, regional, and single cell cDNA analysis within the hippocampus.

    Science.gov (United States)

    Ginsberg, Stephen D; Che, Shaoli

    2004-08-01

    The use of five histochemical stains (cresyl violet, thionin, hematoxylin & eosin, silver stain, and acridine orange) was evaluated in combination with an expression profiling paradigm that included regional and single cell analyses within the hippocampus of post-mortem human brains and adult mice. Adjacent serial sections of human and mouse hippocampus were labeled by histochemistry or neurofilament immunocytochemistry. These tissue sections were used as starting material for regional and single cell microdissection followed by a newly developed RNA amplification procedure (terminal continuation (TC) RNA amplification) and subsequent hybridization to custom-designed cDNA arrays. Results indicated equivalent levels of global hybridization signal intensity and relative expression levels for individual genes for hippocampi stained by cresyl violet, thionin, and hematoxylin & eosin, and neurofilament immunocytochemistry. Moreover, no significant differences existed between the Nissl stains and neurofilament immunocytochemistry for individual CA1 neurons obtained via laser capture microdissection. In contrast, a marked decrement was observed in adjacent hippocampal sections stained for silver stain and acridine orange, both at the level of the regional dissection and at the CA1 neuron population level. Observations made on the cDNA array platform were validated by real-time qPCR using primers directed against beta-actin and glyceraldehyde-3 phosphate dehydrogenase. Thus, this report demonstrated the utility of using specific Nissl stains, but not stains that bind RNA species directly, in both human and mouse brain tissues at the regional and cellular level for state-of-the-art molecular fingerprinting studies.

  11. Gram staining apparatus for space station applications

    Science.gov (United States)

    Molina, T. C.; Brown, H. D.; Irbe, R. M.; Pierson, D. L.

    1990-01-01

    A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space.

  12. Research on pre-staining gel electrophoresis

    International Nuclear Information System (INIS)

    Zhong Ruibo; Liu Yushuang; Zhang Ping; Liu Jingran; Zhao Guofen; Zhang Feng

    2014-01-01

    Background: Gel electrophoresis is a powerful biochemical separation technique. Most biological molecules are completely transparent in the visible region of light, so it is necessary to use staining to show the results after gel electrophoresis, and the general steps of conventional staining methods are time-consuming. Purpose: We try to develop a novel approach to simplify the gel electrophoresis: Pre-Staining Gel Electrophoresis (PSGE), which can make the gel electrophoresis results monitored in real time. Methods: Pre-stain the protein samples with Coomassie Brilliant Blue (CBB) for 30 min before loading the sample into the gel well. Results and Conclusion: PSGE can be successfully used to analyze the binding efficiency of Bovine Serum Albumin (BSA) and amphiphilic polymer via chemical coupling and physical absorption, and the double PSGE also shows a great potential in bio-analytical chemistry. (authors)

  13. In-situ chemical analyses of trans-polyisoprene by histochemical staining and Fourier transform infrared microspectroscopy in a rubber-producing plant, Eucommia ulmoides Oliver.

    Science.gov (United States)

    Bamba, Takeshi; Fukusaki, Ei-Ichiro; Nakazawa, Yoshihisa; Kobayashi, Akio

    2002-10-01

    The localization of polyisoprene in young stem tissues of Eucommia ulmoides Oliver was investigated by histochemical staining and Fourier transform infrared (FT-IR) microspectroscopy. The fibrous structures were stained with Oil Red O. FT-IR microspectroscopic analysis proved that the fibrous structures were trans-polyisoprene. Granular structures stained with the dye, and characteristic absorptions at 2,960 cm(-1) and 1,430 cm(-1) in FT-IR suggested that trans-polyisoprene accumulated in the vicinity of the cambium layer. We have thus successfully shown for the first time the localization of trans-polyisoprene in plant tissues, and our histological investigation allowed us to presume the main sites of biosynthesis and accumulation of trans-rubber. Furthermore, a new technical approach, the preparation of sections using an electronic freezing unit and the in situ analysis of polyisoprene using FT-IR microspectroscopy, is demonstrated to be a promising method for determining the accumulation of polyisoprene as well as other metabolites.

  14. Syphilitic lymphadenitis clinically and histologically mimicking lymphogranuloma venereum.

    Science.gov (United States)

    Wessels, Annesu; Bamford, Colleen; Lewis, David; Martini, Markus; Wainwright, Helen

    2016-04-19

    An inguinal lymph node was discovered incidentally during surgery for a suspected strangulated inguinal hernia. The patient had recently been treated for candidal balanoposthitis and was known to have a paraphimosis. A new foreskin ulcer was discovered when he was admitted for the hernia surgery. The lymph node histology showed stellate abscesses suggestive of lymphogranuloma venereum (LGV). Chlamydial serologic tests were negative. As the histological appearance and clinical details provided were thought to suggest LGV, tissue was also sent for a real-time quadriplex polymerase chain reaction assay. This was used to screen for Chlamydia trachomatis in conjunction with other genital ulcer-related pathogens. The assay was negative for C. trachomatis, but positive for Treponema pallidum. Further histochemical staining of the histological specimen confirmed the presence of spirochaetes.

  15. Propidium iodide (PI) stains Nissl bodies and may serve as a quick marker for total neuronal cell count.

    Science.gov (United States)

    Niu, Junfei; Li, Chunman; Wu, Haihui; Feng, Xianling; Su, Qingning; Li, Shihe; Zhang, Lihong; Yew, David Tai Wai; Cho, Eric Yu Pang; Sha, Ou

    2015-03-01

    Propidium iodide (PI) reacts with both DNA and RNA and is a commonly used fluorescent reagent for nucleic acid staining. The aim of the study was to compare the cellular staining patterns of PI with that of Nissl staining in rat nervous tissues and to report a modified staining method that selectively labels Nissl bodies in neurons. Cryosections and paraffin sections of different tissues of normal Sprague-Dawley rats, including trigeminal ganglia, dorsal root ganglia, spinal cord, liver, and small intestine, were stained by either PI or the hematoxylin and eosin method. Some sections were treated with RNase or DNase before the above staining, and some were double stained with PI and a Nissl stain. The sections were observed by light, fluorescence or confocal microscopy. Results showed strong PI signals detected as patterns of granules in the neuronal cytoplasm of all nervous tissues, whereas the staining of neuronal nuclei was weaker. In contrast, nuclei of neuroglial cells were strongly stained by PI, while the cytoplasm was not obviously stained. Pretreatment of the neural tissue with RNase abolished the PI signals. Furthermore, the PI positive granules in neuronal cytoplasm co-localized with Nissl bodies stained by the fluorescent Nissl stain. When the tissue was pretreated with DNase, PI only stained the cytoplasmic granules of neurons, but not that of glial cells. Our results show that PI stains Nissl bodies and may serve as an economical and convenient neuron marker for neuronal cell counting when specific neural markers such as antibodies are not readily available. Copyright © 2015. Published by Elsevier GmbH.

  16. Secondary syphilis in HIV positive individuals: correlation with histopathologic findings, CD4 counts, and quantity of treponemes in microscopic sections.

    Science.gov (United States)

    Rosa, Gabriela; Procop, Gary W; Schold, Jesse D; Piliang, Melissa P

    2016-10-01

    Although syphilis is uncommon, infection rates are much higher in HIV-infected individuals than the general population. A proposed explanation is impaired cellular immunity with HIV infection. A search of one institution yielded 10 patients with a diagnosis of secondary syphilis on skin biopsy, positive syphilis serology and available CD4 counts. We evaluated 11 biopsies from the 10 patients. We correlated the patients' CD4 counts with the histologic findings and with the number of treponemes on skin biopsies, highlighted by immunohistochemistry (IHC). We also compared the detection of spirochetes in silver stained sections (e.g. Warthin-Starry) with T. pallidum IHC. All biopsies were assessed for various histologic features. The sensitivity of IHC to detect treponemes was 64% and of silver stain was 9% (p-value 0.04). The number of treponemes on the biopsies was determined by IHC. High numbers of spirochetes (i.e. >100 per 10 hpf) were only seen in patients with CD4 counts less than 250 cells/ml. The most consistent histologic finding was a moderate to severe lymphoplasmacytic infiltrate. Although the study is small, it appears that a higher number of spirochetes is associated with CD4 counts less than 250 cell/ml. The T. pallidum IHC stain was vastly superior to the Warthin-Starry stain. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. DRAQ5 and Eosin ('D&E') as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues.

    Science.gov (United States)

    Elfer, Katherine N; Sholl, Andrew B; Wang, Mei; Tulman, David B; Mandava, Sree H; Lee, Benjamin R; Brown, J Quincy

    2016-01-01

    Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA), is too time- and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E") enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis. These results indicate

  18. DRAQ5 and Eosin ('D&E' as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues.

    Directory of Open Access Journals (Sweden)

    Katherine N Elfer

    Full Text Available Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA, is too time- and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E" enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis

  19. Evidence of specialized tissue in human interatrial septum: histological, immunohistochemical and ultrastructural findings.

    Science.gov (United States)

    Mitrofanova, Lubov B; Gorshkov, Andrey N; Lebedev, Dmitry S; Mikhaylov, Evgeny N

    2014-01-01

    There is a paucity of information on structural organization of muscular bundles in the interatrial septum (IAS). The aim was to investigate histologic and ultrastructural organization of muscular bundles in human IAS, including fossa ovalis (FO) and flap valve. Macroscopic and light microscopy evaluations of IAS were performed from postmortem studies of 40 patients. Twenty three IAS specimens underwent serial transverse sectioning, and 17--longitudinal sectioning. The transverse sections from 10 patients were immunolabeled for HCN4, Caveolin3 and Connexin43. IAS specimens from 6 other patients underwent electron microscopy. In all IAS specimens sections the FO, its rims and the flap valve had muscle fibers consisting of working cardiac myocytes. Besides the typical cardiomyocytes there were unusual cells: tortuous and horseshoe-shaped intertangled myocytes, small and large rounded myocytes with pale cytoplasm. The cells were aggregated in a definite structure in 38 (95%) cases, which was surrounded by fibro-fatty tissue. The height of the structure on transverse sections positively correlated with age (P = 0.03) and AF history (P = 0.045). Immunohistochemistry showed positive staining of the cells for HCN4 and Caveolin3. Electron microscopy identified cells with characteristics similar to electrical conduction cells. Specialized conduction cells in human IAS have been identified, specifically in the FO and its flap valve. The cells are aggregated in a structure, which is surrounded by fibrous and fatty tissue. Further investigations are warranted to explore electrophysiological characteristics of this structure.

  20. Evidence of specialized tissue in human interatrial septum: histological, immunohistochemical and ultrastructural findings.

    Directory of Open Access Journals (Sweden)

    Lubov B Mitrofanova

    Full Text Available There is a paucity of information on structural organization of muscular bundles in the interatrial septum (IAS. The aim was to investigate histologic and ultrastructural organization of muscular bundles in human IAS, including fossa ovalis (FO and flap valve.Macroscopic and light microscopy evaluations of IAS were performed from postmortem studies of 40 patients. Twenty three IAS specimens underwent serial transverse sectioning, and 17--longitudinal sectioning. The transverse sections from 10 patients were immunolabeled for HCN4, Caveolin3 and Connexin43. IAS specimens from 6 other patients underwent electron microscopy.In all IAS specimens sections the FO, its rims and the flap valve had muscle fibers consisting of working cardiac myocytes. Besides the typical cardiomyocytes there were unusual cells: tortuous and horseshoe-shaped intertangled myocytes, small and large rounded myocytes with pale cytoplasm. The cells were aggregated in a definite structure in 38 (95% cases, which was surrounded by fibro-fatty tissue. The height of the structure on transverse sections positively correlated with age (P = 0.03 and AF history (P = 0.045. Immunohistochemistry showed positive staining of the cells for HCN4 and Caveolin3. Electron microscopy identified cells with characteristics similar to electrical conduction cells.Specialized conduction cells in human IAS have been identified, specifically in the FO and its flap valve. The cells are aggregated in a structure, which is surrounded by fibrous and fatty tissue. Further investigations are warranted to explore electrophysiological characteristics of this structure.

  1. Lectins stain cells differentially in the coral, Montipora capitata

    Science.gov (United States)

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  2. Calcium hydroxylapatite treatment of human skin: evidence of collagen turnover through picrosirius red staining and circularly polarized microscopy

    Directory of Open Access Journals (Sweden)

    Zerbinati N

    2018-01-01

    Full Text Available Nicola Zerbinati,1 Alberto Calligaro2 1Department of Surgical and Morphological Sciences, University of Insubria (Varese and Polyspecialist Medical Center, Pavia, 2Department of Public Health, Experimental and Forensic Medicine, Unit of Histology and Embryology, University of Pavia, Pavia, Italy Background: Calcium hydroxylapatite (CaHA, Radiesse® is a biocompatible, injectable filler for facial soft-tissue augmentation that provides volume to tissues, followed by a process of neocollagenesis for improved skin quality. Objective: To examine the effects of CaHA treatment on the molecular organization of collagen using a combination of picrosirius red staining and circularly polarized light microscopy.Methods: Five subjects received subdermal injection of 0.3 mL of CaHA in tissues scheduled for removal during abdominoplasty 2 months later. Tissue specimens from the CaHA injection site and a control untreated area were obtained from excised skin at the time of surgery. Processed tissue sections were stained with picrosirius red solution 0.1% and visualized under circularly polarized light microscopy for identification of thick mature (type I and thin newly formed (type III collagen fibers. Pixel signals from both the control and CaHA-treated areas were extracted from the images, and morphometric computerized hue analysis was performed to provide a quantitative evaluation of mature and newly formed collagen fibers.Results: Under picrosirius red staining and circularly polarized light microscopy, green/yellow areas (thin newly formed collagen type III were visible among the collagen fibers in tissue sections from the area of CaHA injection. In contrast, the majority of the collagen fibers appeared red (thick mature collagen type I in control tissues. Morphometric analysis confirmed that, following CaHA treatment, the proportion of fibers represented by thin newly formed collagen type III increased significantly (p<0.01 in comparison with the

  3. Selection and application of exterior stains for wood

    Science.gov (United States)

    R. Sam. Williams; William C. Feist

    1999-01-01

    Exterior stains for wood protect the wood surface from sunlight and moisture. Because stains are formulated to penetrate the wood surface, they are not prone to crack or peel as can film-forming finishes, such as paints. This publication describes the properties of stains and wood, methods for applying stains, and the expected service life of stains.

  4. Staining Methods for Normal and Regenerative Myelin in the Nervous System.

    Science.gov (United States)

    Carriel, Víctor; Campos, Antonio; Alaminos, Miguel; Raimondo, Stefania; Geuna, Stefano

    2017-01-01

    Histochemical techniques enable the specific identification of myelin by light microscopy. Here we describe three histochemical methods for the staining of myelin suitable for formalin-fixed and paraffin-embedded materials. The first method is conventional luxol fast blue (LFB) method which stains myelin in blue and Nissl bodies and mast cells in purple. The second method is a LBF-based method called MCOLL, which specifically stains the myelin as well the collagen fibers and cells, giving an integrated overview of the histology and myelin content of the tissue. Finally, we describe the osmium tetroxide method, which consist in the osmication of previously fixed tissues. Osmication is performed prior the embedding of tissues in paraffin giving a permanent positive reaction for myelin as well as other lipids present in the tissue.

  5. Influence of Helicobacter pylori Colonization on Histological Grading of Chronic Gastritis in Korean Patients with Peptic Ulcer

    Science.gov (United States)

    Park, Joongwon; Kim, Mi Kyung; Park, Sill Moo

    1995-01-01

    Objectives: We conducted an analysis of correlation between histological grading of chronic gastritis and the presence of H. pylori infection to investigate if H. pylori influences histological severity of chronic gastritis in Korean patients with peptic ulcers. Methods: Gastroscopic antral biopsy specimens and peripheral venous blood were taken from 80 patients with gastric or duodenal ulcers. H. pylori was identified microscopically in sections with Giemsa staining and quantitative grading of cultured H. pylori was reported on a scale 0 to 3. The histopathological features of biopsy specimens were reported according to the Sydney classification of chronic gastritis. Serum gastritis and pepsinogen concentrations were measured by radioimmunoassay. Results: H. pylori was identified in 62.5% (20 of 32 GU, 30 of 48 DU) of the study group. Gastric clonization rate of H. pylori did not increased with age. Forty of 50 biopsy specimens with H. pylori and also 23 of 30 biopsy specimens without H. pylori showed active chronic gastritis. There was no significant correlation overall between the presence of H. pylori and histological grading of chronic gastritis, including activity, and also no association was found between the quantitative grading of H. pylori and the histological grading of chronic gastritis. With and without H. pylori, a mean of serum gastritis concentration (79.4±43.0 pg/ml and 80.2±31.9 pg/ml) showed no significant difference, but a mean of serum pepsinogen concentration (87.7±41.6 ng/ml and 119±34.4 ng/ml) showed significant difference between the populations with and without H. pylori (p=0.001) Conclusions: The influence of H. pylori on histological grading of chronic gastritis in Korean is less than that in prior studies of Western countries, and further investigation of pathogenesis of H. pylori in chronic gastritis and peptic ulceration is necessary. PMID:7495770

  6. Histological grading patterns in patients of cutaneous leishmaniasis

    International Nuclear Information System (INIS)

    Saleem, K.; Ayaz, B.; Shaikh, A.

    2007-01-01

    To determine the histological grading patterns in a cohort of hospitalized patients of cutaneous leishmaniasis. One hundred patients of Cutaneous Leishmaniasis (CL), admitted in dermatology wards at PNS Shifa Hospital, Karachi, were examined. Only admitted patients of all ages and both sexes were included in the study. Patients of CL, who had received or were receiving systemic treatment were excluded. The lesions having marked secondary bacterial infection were also excluded. Initial diagnosis was clinical. History of being to an endemic area supported the diagnosis. The lesions were divided in two groups. Early, with duration less than 03 months and late, with duration between 3 and 12 months. The clinical lesions were noted as nodules, plaques, ulcers, crusted ulcers, lupoid lesions and plaques with scarring. Three types of skin smears (slit skin smear, saline aspirate smear and dab smear) were taken and examined with Giemsa stain. Cultures were performed on Nicolle-Novy-MacNeal (NNN) culture medium from Defense Scientific and Technology Organization (DESTO) Lab., Pakistan. Incisional skin biopsies were done. The biopsy specimens were examined by hemotoxylin and eosin stain (H and E stain). The number of Leishmania Tropica (LT) bodies was graded according to modified Ridley's parasitic index 1983. Clinical features were correlated with the histological patterns. Five histological patterns were identified in current study: 1) diffuse dermal infiltration without necrosis, 2) patchy dermal infiltration, 3) diffuse dermal infiltration with necrosis, 4) early reactive granuloma formation and 5) established epithelioid granuloma formation. LT bodies were identified in 75% of cases. Epidermal features were non-specific. The early lesions presented with diffuse infiltrate and late lesions showed granuloma formation. Five distinct types of histological patterns of CL have been recognized in this study. The early lesions presented with diffuse infiltrate and late lesions

  7. Intrastromal Injection of China Painting Ink in Corneas of Male Rabbits: Clinical and Histological Study.

    Science.gov (United States)

    Alsmman Hassan, Alahmady Hamad; Abd Elhaliem Soliman, Nesreen Gamal-Eldeen

    2016-01-01

    Background. Many patients with corneal opacity or complicated cataract in blind eye ask for cosmoses. In this study we tried to investigate the staining of corneas of male rabbits by Rotring China painting ink and to study the histological changes. Method. 10 eyes of 10 male Baladi Egyptian rabbits were injected (0.1 mL) intrastromally in the cornea by the use of China painting ink (Rotring Tinta China) through insulin syringe (27-gauge needle) by single injection; clinical follow-up is for 6 months and lastly the rabbits were scarified and the stained eyes were enucleated for histological analysis. Results. Clinically the stain was stable in color and distribution in corneas with no major complications. Histological results of the stained rabbit corneas showed blackish pigmentation in the corneal stroma without any inflammatory cellular infiltration. Some fibroblast cells had pigment granules in their cytoplasm in the adjacent layers. Conclusion. Corneal staining by China painting ink is effective and safe in staining of male rabbits cornea; however further study in human corneas with longer follow-up period is advisable.

  8. Intrastromal Injection of China Painting Ink in Corneas of Male Rabbits: Clinical and Histological Study

    Directory of Open Access Journals (Sweden)

    Alahmady Hamad Alsmman Hassan

    2016-01-01

    Full Text Available Background. Many patients with corneal opacity or complicated cataract in blind eye ask for cosmoses. In this study we tried to investigate the staining of corneas of male rabbits by Rotring China painting ink and to study the histological changes. Method. 10 eyes of 10 male Baladi Egyptian rabbits were injected (0.1 mL intrastromally in the cornea by the use of China painting ink (Rotring Tinta China through insulin syringe (27-gauge needle by single injection; clinical follow-up is for 6 months and lastly the rabbits were scarified and the stained eyes were enucleated for histological analysis. Results. Clinically the stain was stable in color and distribution in corneas with no major complications. Histological results of the stained rabbit corneas showed blackish pigmentation in the corneal stroma without any inflammatory cellular infiltration. Some fibroblast cells had pigment granules in their cytoplasm in the adjacent layers. Conclusion. Corneal staining by China painting ink is effective and safe in staining of male rabbits cornea; however further study in human corneas with longer follow-up period is advisable.

  9. Comparism of Various Staining Techniques in the Diagnosis of ...

    African Journals Online (AJOL)

    SITWALA COMPUTERS

    external intermediate host, usually an animal, in which sporogenesis and oocyst ... the parasite was detected in 111 of the samples stained,. 100(90.0%) of which .... screen stained slide was the auramine fluorochrome stain. The widely used ...

  10. A quantitative comparison of morphological and histological characteristics of collagen in the rabbit medial collateral ligament.

    Science.gov (United States)

    Wan, Chao; Hao, Zhixiu; Wen, Shizhu

    2013-12-01

    Collagen fiber is one of the critical factors in determining mechanical properties of ligaments and both the morphological and histological characteristics of collagen have been widely studied. However, there was still no consensus about whether the morphological characteristics of collagen correlated with its histological characteristics in physiological ligaments. Rabbit medial collateral ligaments (MCLs) were measured under a transmission electron microscope and a polarized light microscope plus picrosirius red-staining to obtain the distributions of collagen fibril diameters and types at different anatomical sites of rabbit MCLs, respectively. The correlation between the fibril diameter and type was determined by a correlation analysis. The collagen fibril diameters at the different anatomical sites had different distributions (unimodal or bimodal) and mean fibril diameters were found to increase significantly from the anterior part to the posterior part (P=0.0482) as well as from the proximal to the distal sections (P=0.0208). Type I collagen in the core portion of MCLs was significantly less than at the other four peripheral areas (P0.05). The low coefficient in the correlation analysis (r=0.3759) demonstrated collagen fibril diameters had no correlation with collagen types. This may provide a new view of collagen types in studying the mechanical behavior of ligaments. Copyright © 2013 Elsevier GmbH. All rights reserved.

  11. Slide-free histology via MUSE: UV surface excitation microscopy for imaging unsectioned tissue (Conference Presentation)

    Science.gov (United States)

    Levenson, Richard M.; Harmany, Zachary; Demos, Stavros G.; Fereidouni, Farzad

    2016-03-01

    Widely used methods for preparing and viewing tissue specimens at microscopic resolution have not changed for over a century. They provide high-quality images but can involve time-frames of hours or even weeks, depending on logistics. There is increasing interest in slide-free methods for rapid tissue analysis that can both decrease turn-around times and reduce costs. One new approach is MUSE (microscopy with UV surface excitation), which exploits the shallow penetration of UV light to excite fluorescent signals from only the most superficial tissue elements. The method is non-destructive, and eliminates requirement for conventional histology processing, formalin fixation, paraffin embedding, or thin sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography optics. MUSE generates diagnostic-quality histological images that can be rendered to resemble conventional hematoxylin- and eosin-stained samples, with enhanced topographical information, from fresh or fixed, but unsectioned tissue, rapidly, with high resolution, simply and inexpensively. We anticipate that there could be widespread adoption in research facilities, hospital-based and stand-alone clinical settings, in local or regional pathology labs, as well as in low-resource environments.

  12. The Evaluation of Epiphyseal Plate Histological Changes in Osteopetrotic op/op Mice.

    Directory of Open Access Journals (Sweden)

    Aligholi Sobhani

    2003-09-01

    Full Text Available This study was designed for evaluation of epiphyseal plate histological changes of femur bones in osteopetrotic op/op mice.In this study 5 osteopetrotic op/op mice which were purchased from the commercial source were used.The animals were killed by overdose of chloroform and their femur bones were extracted. The bones were fixed in 10% formaldehyde and decalcified by HCl (0.6N, and routine histological processing were performed. The sections were stained by H&E methods and studied by conventional light microscopy. The results showed that, proliferative zone (PZ and especially hypertrophic zone (HZ were much thickened. In the ossification zone, trabecular bones were irregular and atypical osteoblast cells were observed. The osteoclast cells were not attached to trabecular bones. The bone marrow cavity was restricted and bone marrow cells were poor and scattered. Findings of the present investigation are similar to those reported about epiphyseal plate in osteosclerotic (OC mice in which epiphyseal plate especially hypertrophic zone was thickened and chondrocytes were not substituted for osteoblasts in calcified cartilage area. Also, osteoclast cells had been inactive or absent in OC mice. For prevention of other complication due to the epiphyseal plate changes in new borne, suitable and punctually treatment protocols such as prescription of Macrophage Colony Stimulating-Factor (MCS-F could be useful.

  13. Mobile phone radiation during pubertal development has no effect on testicular histology in rats.

    Science.gov (United States)

    Tumkaya, Levent; Kalkan, Yildiray; Bas, Orhan; Yilmaz, Adnan

    2016-02-01

    Mobile phones are extensively used throughout the world. There is a growing concern about the possible public health hazards posed by electromagnetic radiation emitted from mobile phones. Potential health risk applies particularly to the most intensive mobile phone users-typically, young people. The aim of this study was to investigate the effects of mobile phone exposure to the testes, by assessing the histopathological and biochemical changes in the testicular germ cells of rats during pubertal development. A total of 12 male Sprague Dawley rats were used. The study group (n = 6) was exposed to a mobile phone for 1 h a day for 45 days, while the control group (n = 6) remained unexposed. The testes were processed with routine paraffin histology and sectioned. They were stained with hematoxylin-eosin, caspase 3, and Ki-67 and then photographed. No changes were observed between the groups (p > 0.05). The interstitial connective tissue and cells of the exposed group were of normal morphology. No abnormalities in the histological appearance of the seminiferous tubules, including the spermatogenic cycle stage, were observed. Our study demonstrated that mobile phones with a low specific absorption rate have no harmful effects on pubertal rat testicles. © The Author(s) 2013.

  14. Histological scoring and associated risk factors of non-alcoholic fatty liver disease.

    Science.gov (United States)

    Majid, N; Ali, Z; Rahman, M R; Akhter, A; Rajib, R C; Ahmad, F; Sharmin, S; Akond, A K; Huq, N

    2013-10-01

    Non alcoholic steatohepatitis is a hepatic disorder with histological features of alcohol induced liver disease that occurs in individual who do not consume significant alcohol. Liver biopsy is an important part of the evaluation in term of both grade & stage. A cross sectional study was carried out in the department of Pathology, Dhaka Medical College, Dhaka & department of Hepatology, Bangabandhu Sheikh Mujib Medical University (BSMMU) from July 2007 to June 2009. Total 55 adult subjects of both sex were included on the basis of predefined inclusion & exclusion criteria in this study to evaluate the histological pattern of non alcoholic fatty liver disease (NAFLD) and its correlation with risk factors. Liver biopsy was done and H & E and Masson's Trichrome stain slides were examined to evaluate the grade and stage of NAFLD. Scoring and semiquantitative assessment of steatosis and NAFLD severity was done according to Kleiner scale known as NAFLD activity score (NAS). The results of Pearson correlation showed only BMI and triglyceride level significantly correlated with NAS score. The results of Spearman's rank correlation showed that BMI, central obesity, triglyceridaemia and age significantly correlated with staging of fibrosis. The results of multiple regression analysis showed that variation of NAS depend on BMI and triglyceride level. The study also revealed that risk factors contributed about 29% risk for the occurrence of non alcoholic steatohepatitis.

  15. The anterior cruciate ligament-lateral meniscus complex: A histological study.

    Science.gov (United States)

    Furumatsu, Takayuki; Kodama, Yuya; Maehara, Ami; Miyazawa, Shinichi; Fujii, Masataka; Tanaka, Takaaki; Inoue, Hiroto; Ozaki, Toshifumi

    2016-01-01

    The anterior root of the lateral meniscus (LM) dives underneath the tibial attachment of the anterior cruciate ligament (ACL). Although the distinct role of meniscal attachments has been investigated, the relationship between the LM anterior insertion (LMAI) and ACL tibial insertion (ACLTI) remains unclear. This study histologically analyzed the LMAI and ACLTI. Samples were divided into four regions in an anterior-to-posterior direction. Histological measurements of these insertion sites were performed using safranin O-stained coronal sections. Distribution and signal densities of type I and II collagen were quantified. The ACLTI and LMAI formed the ACL-LM complex via fiber connections. The anterior part of the ACLTI had a widespread attachment composed of dense fibers. Attachment fibers of the LMAI became dense and wide gradually at the middle-to-posterior region. The ACL-LM transition zone (ALTZ) was observed between the LMAI and the lateral border of the ACLTI at the middle part of the ACL tibial footprint. Type II collagen density of the LMAI was higher than that of the ACLTI and ALTZ. Our results can help create an accurate tibial bone tunnel within the dense ACL attachment during ACL reconstruction surgery.

  16. Reconsidering the H&E stain as the gold standard in assessing the depth of burn wounds.

    Science.gov (United States)

    Rosenberg, Arlene S

    2017-12-01

    While histological examination is considered by most as the gold standard for burn depth assessment, it has no practical use in the clinical setting. It has, however, been used in the research setting, as a mean for evaluating emerging techniques of depth measurement. Due to the limitations of the H&E stain, other stains have also been explored, such as lactate dehydrogenase (LDH), as presented in this issue, in "Improving the Histologic Characterization of Burn Depth." As the determination of burn depth is not a typical subject in dermatopathology, a summary of selected techniques and the possible role for the LDH stain in future research, is described herein. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Characterization of paint layers and stained glasses

    International Nuclear Information System (INIS)

    Biagi Maino, D.; Ciancabilla, L.; Gandolfi, G.; Maino, G.; Bruni, S.; Ferriani, S.; Visparelli, D.

    2000-01-01

    Due to the higher magnification with respect to traditional optical microscopes, the scanning electron microscopy (SEM) has been extensively applied in recent years to the investigation of elemental composition of many different types of artistic objects. The back-scattered and secondary electrons produced when the SEM electron beam hits the sample can be detected and converted in electronic signals which give rise to images of the scanned area. These images can be recorded in digital format and stored on a computer for subsequent processing. Moreover, in addition to the back-scattered and secondary electrons, the impact of the electron beam on the sample produces a X-ray spectrum, which can be further processed and analysed using an X-ray spectrometer coupled to the SEM. Therefore, it is possible to yield the chemical composition of the sample, analogously to the X-ray fluorescence analysis. Moreover, the transmission electron microscopy (TEM) can be applied to the characterization of paint layers, making ultra-thin sections in which the paint and ground layers are preserved intact. In comparison with usual SEM measurements, the TEM technique is more precise, because of the higher spatial resolution in both the microanalysis and diffraction modes, of the order of 10-20 nm. This precision allows unique identification of each component in the layer and determination of the crystallographic structure, thus characterizing even the smallest particles of each pigment and pointing out minor components. It is then possible to establish whether a pigment is natural, manufactured, its origin as well as approximate datations. In this work, we describe the research activities performed in the laboratory recently established at the ENEA (Italian National Agency for New Technology, Energy and Environment) Applied Physics Division, where dedicated SEM and TEM are connected by a suitable imaging system to a powerful computing system for image acquisition and processing. Use has

  18. Pleural and Pulmonary Staining at Inferior Phrenic Arteriography Mimicking a Tumor Staining of Hepatocellular Carcinoma

    International Nuclear Information System (INIS)

    Lee, Deok Hee; Hwang, Jae Cheol; Lim, Soo Mee; Yoon, Hyun-Ki; Sung, Kyu-Bo; Song, Ho-Young

    2000-01-01

    Purpose: To describe the findings of pleural and pulmonary staining of the inferior phrenic artery, which can be confused with tumor staining during transarterial chemoembolization (TACE) of hepatoma.Methods: Fifteen patients who showed pleural and pulmonary staining without relationship to hepatic masses at inferior phrenic arteriography were enrolled. The staining was noted at initial TACE (n = 8), at successive TACE (n = 5), and after hepatic surgery (n = 2). The angiographic pattern, the presence of pleural change on computed tomography (CT), and clinical history were evaluated.Results: Draining pulmonary veins were seen in all cases. The lower margin of the staining corresponded to the lower margin of the pleura in 10 patients. CT showed pleural and/or pulmonary abnormalities in all cases. After embolization of the inferior phrenic artery, the accumulation of iodized oil in the lung was noted.Conclusion: Understanding the CT and angiographic findings of pleural and pulmonary staining during TACE may help differentiate benign staining from tumor staining

  19. CD8+ T cell infiltration in breast and colon cancer: A histologic and statistical analysis.

    Directory of Open Access Journals (Sweden)

    James Ziai

    Full Text Available The prevalence of cytotoxic tumor infiltrating lymphocytes (TILs has demonstrated prognostic value in multiple tumor types. In particular, CD8 counts (in combination with CD3 and CD45RO have been shown to be superior to traditional UICC staging in colon cancer patients and higher total CD8 counts have been associated with better survival in breast cancer patients. However, immune infiltrate heterogeneity can lead to potentially significant misrepresentations of marker prevalence in routine histologic sections. We examined step sections of breast and colorectal cancer samples for CD8+ T cell prevalence by standard chromogenic immunohistochemistry to determine marker variability and inform practice of T cell biomarker assessment in formalin-fixed, paraffin-embedded (FFPE tissue samples. Stained sections were digitally imaged and CD8+ lymphocytes within defined regions of interest (ROI including the tumor and surrounding stroma were enumerated. Statistical analyses of CD8+ cell count variability using a linear model/ANOVA framework between patients as well as between levels within a patient sample were performed. Our results show that CD8+ T-cell distribution is highly homogeneous within a standard tissue sample in both colorectal and breast carcinomas. As such, cytotoxic T cell prevalence by immunohistochemistry on a single level or even from a subsample of biopsy fragments taken from that level can be considered representative of cytotoxic T cell infiltration for the entire tumor section within the block. These findings support the technical validity of biomarker strategies relying on CD8 immunohistochemistry.

  20. A comparative assessment of commonly employed staining ...

    African Journals Online (AJOL)

    Following an increase in the number of reports of Cryptosporidium infections and the problems encountered in detecting these organisms in faecal smears, a comparative assessment of a modification of the Sheather's flotation technique and other commonly employed staining procedures proved the modified Sheather's ...

  1. Photoacoustic imaging of port-wine stains

    NARCIS (Netherlands)

    Kolkman, Roy G. M.; Mulder, Miranda J.; Glade, Conrad P.; Steenbergen, Wiendelt; van Leeuwen, Ton G.

    2008-01-01

    BACKGROUND AND OBJECTIVE: To optimize laser therapy of port-wine stains (PWSs), information about the vasculature as well as lesion depth is valuable. In this study we investigated the use of photoacoustic imaging (PAI) to obtain this information. STUDY DESIGN/MATERIALS AND METHODS: PAI uses pulsed

  2. Photoacoustic Imaging of Port-Wine Stains

    NARCIS (Netherlands)

    Kolkman, R.G.M.; Mulder, M.J.; Mulder, Miranda J.; Glade, Conrad P.; Steenbergen, Wiendelt; van Leeuwen, Ton

    2008-01-01

    Background and Objective: To optimize laser therapy of port-wine stains (PWSs), information about the vasculature as well as lesion depth is valuable. In this study we investigated the use of photoacoustic imaging (PAI) to obtain this information. - Study Design/Materials and Methods: PAI uses

  3. Histological image classification using biologically interpretable shape-based features

    International Nuclear Information System (INIS)

    Kothari, Sonal; Phan, John H; Young, Andrew N; Wang, May D

    2013-01-01

    Automatic cancer diagnostic systems based on histological image classification are important for improving therapeutic decisions. Previous studies propose textural and morphological features for such systems. These features capture patterns in histological images that are useful for both cancer grading and subtyping. However, because many of these features lack a clear biological interpretation, pathologists may be reluctant to adopt these features for clinical diagnosis. We examine the utility of biologically interpretable shape-based features for classification of histological renal tumor images. Using Fourier shape descriptors, we extract shape-based features that capture the distribution of stain-enhanced cellular and tissue structures in each image and evaluate these features using a multi-class prediction model. We compare the predictive performance of the shape-based diagnostic model to that of traditional models, i.e., using textural, morphological and topological features. The shape-based model, with an average accuracy of 77%, outperforms or complements traditional models. We identify the most informative shapes for each renal tumor subtype from the top-selected features. Results suggest that these shapes are not only accurate diagnostic features, but also correlate with known biological characteristics of renal tumors. Shape-based analysis of histological renal tumor images accurately classifies disease subtypes and reveals biologically insightful discriminatory features. This method for shape-based analysis can be extended to other histological datasets to aid pathologists in diagnostic and therapeutic decisions

  4. Correlation between ICDAS and histology: Differences between stereomicroscopy and microradiography with contrast solution as histological techniques.

    Directory of Open Access Journals (Sweden)

    Samara de Azevedo Gomes Campos

    Full Text Available Detection of occlusal caries with visual examination using ICDAS correlates strongly with histology under stereomicroscopy (SM, but dentin aspects under SM are ambiguous regarding mineral content. Thus, our aim was to test two null hypotheses: SM and microradiography result in similar correlations between ICDAS and histology; SM and microradiography result in similar positive (PPV and negative predictive values (NPV of ICDAS cut-off 1-2 (scores 0-2 as sound with histological threshold D3 (demineralization in the inner third of dentin. Occlusal surfaces of extracted permanent teeth (n = 115 were scored using ICDAS. Undemineralized ground sections were histologically scored using both SM without contrast solution and microradiography after immersion in Thoulet's solution 1.47 for 24 h (MRC. Correlation between ICDAS and histology differed from SM (0.782 to MRC (0.511 (p = 0.0002, with a large effect size "q" of 0.49 (95% CI: 0.638/0.338. For ICDAS cut-off 1-2 and D3, PPV from MRC (0.56 was higher than that from SM (0.28 (p< 0.00001; effect size h = 0.81, and NPV from MRC (0.72 was lower than that from SM (1,00 (p < 0.00001; effect size h = 1.58. In conclusion, SM overestimated the correlation between ICDAS and lesion depth, and underestimated the number of occlusal surfaces with ICDAS cut-off 1-2 and deep dentin demineralization.

  5. Effectiveness of Vascular Markers (Immunohistochemical Stains) in Soft Tissue Sarcomas.

    Science.gov (United States)

    Naeem, Namra; Mushtaq, Sajid; Akhter, Noreen; Hussain, Mudassar; Hassan, Usman

    2018-05-01

    To ascertain the effectiveness of IHC markers of vascular origin like CD31, CD34, FLI1 and ERG in vascular soft tissue sarcomas including angiosarcomas, Kaposi sarcomas, epithelioid hemangioendothelioma and a non-vascular soft tissue sarcoma (Epithelioid sarcoma). Descriptive study. Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore, from 2011 to 2017. Diagnosed cases of angiosarcomas (n=48), epithelioid hemangioendothelioma (n=9), Kaposi sarcoma (n=9) and epithelioid sarcoma (n=20) were selected. Immunohistochemical staining as performed on formalin fixed paraffin embedded sections. The sections were stained for the following markers: CD34 (VENTANA clone Q Bend 10), CD31 (Leica clone 1 A 10), FLI1 (CELL MARQUE clone MRQ-1) and ERG (CELL MARQUE clone EP111). A complete panel of CD34, CD31 and ERG was applied on 8/48 cases of angiosarcomas with triple positivity in 6 cases. Eight cases showed positivity for only CD31 and ERG and 2 cases showed positivity for only ERG. A complete panel of CD34, CD31 and ERG was applied on 3/9 cases of epithelioid hemangioendothelioma with positivity for all markers in 2 cases. Combined positivity for ERG and CD34 was seen in 2 cases and on 4 cases only CD31 immunohistochemical was solely applied with 100% positivity. FLI1 was not applied on any case. Among 9 cases of Kaposi sarcoma, ERG, CD34 and CD31 in combination were applied on only 1 case with triple positivity. Remaining cases show positivity for either CD34, CD31 or FLI1. Majority of cases of epithelioid sarcomas were diagnosed on the basis of cytokeratin and CD34 positivity with loss of INI1. The other vascular markers showed negativity in all cases. Among these four markers, ERG immunohistochemical stain is highly effective for endothelial differentiation due to its specific nuclear staining pattern in normal blood vessel endothelial cells (internal control) as well as neoplastic cells of vascular tumors and lack of background staining.

  6. Histological evaluation of thyroid lesions using a scanning acoustic microscope

    Directory of Open Access Journals (Sweden)

    Miura K

    2014-02-01

    Full Text Available Katsutoshi Miura,1 Hiroyuki Mineta2 1Department of Health Science, Pathology, and Anatomy, 2Department of Otorhinolaryngology, Hamamatsu University School of Medicine, Hamamatsu, Japan Purpose: A scanning acoustic microscope (SAM uses an ultrasound to image an object by plotting the speed-of-sound (SOS through tissues on screen. Because hard tissues result in great SOS, SAM can provide data on the tissue elasticity. This paper investigated the utility of SAM in evaluating thyroid lesions. Methods: Formalin-fixed, paraffin sections were scanned with a 120 MHz transducer. SOS through each area was calculated and plotted on the screen to provide histological images, and SOS of each lesion was compared and statistically analyzed. Results: High-concentrated colloids, red blood cells, and collagen fibers showed great SOS, while low-concentrated colloids, parathyroids, lymph follicles, and epithelial tissues including carcinomas demonstrated lower SOS. SAM clearly discriminated structure of thyroid components corresponding to low magnification of light microscopy. Thyroid tumors were classified into three groups by average SOS: the fast group consisted of follicular adenomas/carcinomas and malignant lymphomas; the slow group contained poorly differentiated/undifferentiated carcinomas; and the intermediate group comprised papillary/medullary carcinomas. Fragmented colloids, irregular-shaped follicles, and desmoplastic reactions were observed in the invasive area of surrounding carcinomas. Conclusion: The SAM imaging method had the following benefits: 1 precise images were acquired in a few minutes without special staining; 2 structural irregularity and desmoplastic reactions, which indicated malignancy, were detected; 3 images reflected tissue elasticity, which was statistically comparable among lesions by SOS; 4 follicular functional activity was predictable by converting colloid concentration to SOS; and 5 tumor classification was predictable by SOS

  7. Staining plastic blocks with triiodide to image cells and soft tissues in backscattered electron SEM of skeletal and dental tissues

    Directory of Open Access Journals (Sweden)

    A Boyde

    2012-07-01

    Full Text Available Backscattered electron scanning electron microscopy (BSE SEM is an invaluable method for studying the histology of the hard, mineralised components of poly-methyl methacrylate (PMMA or other resin embedded skeletal and dental tissues. Intact tissues are studied in micro-milled or polished block faces with an electron-optical section thickness of the order of a half to one micron and with the area of the section as big as a whole – large or small – bone organ. However, BSE SEM does not give information concerning the distribution of uncalcified, ‘soft’, cellular and extracellular matrix components. This can be obtained by confocal microscopy of the same block and the two sorts of images merged but the blocks have to be studied in two microscope systems. The present work shows a new, simple and economic approach to visualising both components by using the triiodide ion in Lugol's iodine solution to stain the block surface prior to the application of any conductive coating – and the latter can be omitted if charging is suppressed by use of poor vacuum conditions in the SEM sample chamber. The method permits the use of archival tissue, and it will be valuable in studies of both normal growth and development and pathological changes in bones and joints, including osteoporosis and osteoarthritis, and tissue adaptation to implants.

  8. Oil Red O and Hematoxylin and Eosin Staining for Quantification of Atherosclerosis Burden in Mouse Aorta and Aortic Root.

    Science.gov (United States)

    Andrés-Manzano, M Jesús; Andrés, Vicente; Dorado, Beatriz

    2015-01-01

    Methods for staining tissues with Oil Red O and hematoxylin-eosin are classical histological techniques that are widely used to quantify atherosclerotic burden in mouse tissues because of their ease of use, reliability, and the large amount of information they provide. These stains can provide quantitative data about the impact of a genetic or environmental factor on atherosclerotic burden and on the initiation, progression, or regression of the disease, and can also be used to evaluate the efficacy of drugs designed to prevent or treat atherosclerosis. This chapter provides protocols for quantifying atherosclerotic burden in mouse aorta and aortic root, including methods for dissection, Oil Red O staining, hematoxylin-eosin staining, and image analysis.

  9. Laser Treatment of Port Wine Stains

    Science.gov (United States)

    Majaron, Boris; Nelson, J. Stuart

    Port wine stain (PWS), also called nevus flammeus, is a congenital, cutaneous vascular malformation involving post-capillary venules which produce a light pink to red to dark-red-violet discoloration of human skin [1]. PWS occurs in an estimated 3 children per 1000 live births, affecting males and females and all racial groups equally [2]. There appears to be no hereditary predilection for PWS within families. There are no known risk factors or ways to prevent PWS.

  10. Effect of a calcium hydroxide-based root canal dressing on periapical repair in dogs: a histological study.

    Science.gov (United States)

    Leonardo, Mario R; Hernandez, Maria E F T; Silva, Léa A B; Tanomaru-Filho, Mário

    2006-11-01

    To compare the periapical repair of teeth with periapical lesion following root canal treatment by using a calcium hydroxide-based intracanal dressing for several time periods or filling in a single visit. After induction of periapical lesions in 4 dogs, the root canals were prepared using 5.25% sodium hypochlorite for irrigation, and animals were separated into 4 experimental groups; in group I, root canals were filled in a single session; in groups II, III, and IV, a calcium hydroxide-based dressing was kept in place for 15, 30, or 180 days, respectively. Root canals from groups I, II, and III were filled with gutta-percha cones and AH Plus sealer. After 180 days, animals were killed and histological sections were stained with hematoxylin-eosin to evaluate periapical repair. Periapical repair was better in groups II, III, and IV (intracanal dressing) compared with group I (single session; P session treatment.

  11. Investigation of black soot staining in houses

    Energy Technology Data Exchange (ETDEWEB)

    Fugler, D. [Canada Mortgage and Housing Corp., Ottawa, ON (Canada)

    2000-07-01

    Air quality investigators are frequently called upon to determine the origin of streaking, staining or soot marks in both new and old homes. Those marks display common characteristics: black marks along baseboards at interior or exterior walls, behind furniture and at doorways; black smudges on window frames and plastic cabinets; and even shadowing of studs on exterior wall drywall in a few cases. In most instances, carbon soot from a combustion source is the culprit. The combustion sources include furnaces, water heaters, fireplaces, gas dryers, gas ranges, smoking, vehicle exhaust and candle burning. Scepticism about candle soot is prevalent among callers. As a result, a study was initiated in homes where occupants burn candles regularly to investigate soot problems. Samples were collected from five homes, and included stained carpets, filters, and swab samples of black dust or soot. All the houses selected for the study had been built within a three-year period. Some samples of candles commonly burned in those homes were burnt in a laboratory. Air quality audits had been performed in the homes and had revealed other potential pollutant sources. Best practices for cost-effective clean up and control of soot were researched in industry information. The tests conducted in the laboratory found materials consistent with candle soot or residue during microscopic investigations, but no link was established with the stained material obtained from the homes. A few tips for homeowners were included concerning candle burning, and tips for builders were also offered. 1 tab.

  12. Histology of the distal dural ring.

    Science.gov (United States)

    Graffeo, Christopher S; Perry, Avital; Copeland, William R; Raghunathan, Aditya; Link, Michael J

    2017-09-01

    The distal dural ring (DDR) is a conserved intracranial anatomic structure marking the boundary point at which the internal carotid artery (ICA) exits the cavernous sinus (CS) and enters the subarachnoid space. Although the CS has been well described in a range of anatomic studies, to our knowledge no prior study has analyzed the histologic relationship between the ICA and DDR. Correspondingly, our objective was to assess the relationship of the DDR to the ICA and determine whether the DDR can be dissected from the ICA and thus divided, or can only be circumferentially trimmed around the artery. The authors examined ten fresh-frozen, adult cadaveric specimens. A standard frontotemporal craniotomy, orbito-optic osteotomy, and extradural anterior clinoidectomy was performed bilaterally. The cavernous ICA, DDR, and supraclinoid ICA were harvested as an en bloc specimen. Specimens formalin-fixed and paraffin-embedded prior to routine histochemical staining with hematoxylin and eosin and Masson trichrome. In all specimens, marked microscopic investment of the DDR throughout the ICA adventitia was noted. Dural collagen fibers extensively permeated the arterial layers superficial to the muscularis propria, with no evidence of a clear separation between the DDR and arterial adventitia. Histologic analysis suggests that the ICA and DDR are highly interrelated, continuous structures, and therefore attempted intraoperative dissection between these structures may carry an elevated risk of injury to the ICA. We correspondingly recommend careful circumferential trimming of the DDR in lieu of direct dissection in cases requiring mobilization of the clinoidal ICA. Clin. Anat. 30:742-746, 2017. © 2017Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  13. A histologic analysis of the effects of stainless steel and titanium implants adjacent to tendons: an experimental rabbit study.

    Science.gov (United States)

    Nazzal, Adam; Lozano-Calderón, Santiago; Jupiter, Jesse B; Rosenzweig, Jaime S; Randolph, Mark A; Lee, Sang Gil P

    2006-09-01

    The current trend is to treat distal radius fractures with open reduction and internal fixation with either titanium or stainless steel plates. Both provide stable fixation; however, there is minimal evidence concerning the soft-tissue response to these materials. Our objective was to evaluate the response of adjacent extensor tendons to titanium and stainless steel in a rabbit in vivo model and to evaluate the influence of time. Forty rabbits were divided into 5 groups of 8 rabbits each. Groups I and II had unilateral osteotomy of the distal radius followed by dorsal fixation with titanium and stainless steel plates, respectively. Groups III and IV had fixation with titanium and stainless steel, respectively, but without osteotomy. Group V had surgical dissection without osteotomy or plates. Two animals per group were killed at 1, 4, 12, and 24 weeks. The specimens (distal radius, plate, overlying soft tissue, and extensor tendon) were harvested en bloc for histologic analysis. For interface preservation between implant and tissues the specimens were embedded in methylmethacrylate, sectioned, and stained with hematoxylin-eosin. Histologic analysis showed a fibrous tissue layer formed over both implants between the plate and the overlying extensor tendons in the groups treated with plating independently of the material and the presence or absence of osteotomy. This fibrous layer contained the majority of debris. Metallic particles were not observed in the tendon or muscle substance of any animals; however, they were visualized in the tenosynovium. Hematoxylin-eosin-stained sections of groups I through IV showed proliferative fibroblasts and metallic particles; however, this layer was not observed in group V. Statistical analysis did not show differences between the groups regarding the number of cells or metallic particles. Our results indicate that both implants generated adjacent reactive inflammatory tissue and particulate debris. There was no difference in cell

  14. Hematoxylin and eosin stain shows a high sensitivity but sub-optimal specificity in demonstrating iron pigment in liver biopsies.

    Science.gov (United States)

    Alwahaibi, Nasar Yousuf; Alkhatri, Azza Sarhan; Kumar, Johanes Selva

    2015-01-01

    Perls' stain is routinely used to demonstrate iron in liver biopsies. We tested the hypothesis that it may be unnecessary in cases, where no iron or another similar pigment was seen on the routine hematoxylin and eosin (H and E) stained section. The aim of this study was to evaluate the efficiency of H and E stain in demonstrating iron in liver biopsies as well as to determine the possibility of replacing Perls' stain with H and E stain. Two hundred pairs of slides of liver biopsies were taken from the archival files of the Department of Pathology from 2006 to 2011. Perls' and H and E slides were independently reviewed for the presence of iron. Hundred and one cases showed the presence of iron using H and E stain. 84 of 86 cases showed positive iron using both Perls' and H and E stains. Seventeen cases were positive using H and E stain but negative with Perls'. Only two cases did not show the presence of iron using H and E stain. Ninety-seven cases were negative using both Perls' and H and E stains. H and E stain showed a sensitivity, specificity, accuracy, positive predictive valve, and negative predictive value of 97.67%, 85.08%, 90.5%, 83.16%, and 97.98%, respectively. We demonstrate that the H and E stain is a sensitive method to detect iron pigment in liver biopsies, particularly when present in large quantities. A negative H and E stain might obviate the need for extra Perls' staining, thus saving costs and shortening report turn-around times.

  15. Grinding and polishing instead of sectioning for the tissue samples with a graft: Implications for light and electron microscopy.

    Science.gov (United States)

    Mukhamadiyarov, Rinat A; Sevostyanova, Victoria V; Shishkova, Daria K; Nokhrin, Andrey V; Sidorova, Olga D; Kutikhin, Anton G

    2016-06-01

    A broad use of the graft replacement requires a detailed investigation of the host-graft interaction, including both histological examination and electron microscopy. A high quality sectioning of the host tissue with a graft seems to be complicated; in addition, it is difficult to examine the same tissue area by both of the mentioned microscopy techniques. To solve these problems, we developed a new technique of epoxy resin embedding with the further grinding, polishing, and staining. Graft-containing tissues prepared by grinding and polishing preserved their structure; however, sectioning frequently required the explantation of the graft and led to tissue disintegration. Moreover, stained samples prepared by grinding and polishing may then be assessed by both light microscopy and backscattered scanning electron microscopy. Therefore, grinding and polishing outperform sectioning when applied to the tissues with a graft. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Automated robust registration of grossly misregistered whole-slide images with varying stains

    Science.gov (United States)

    Litjens, G.; Safferling, K.; Grabe, N.

    2016-03-01

    Cancer diagnosis and pharmaceutical research increasingly depend on the accurate quantification of cancer biomarkers. Identification of biomarkers is usually performed through immunohistochemical staining of cancer sections on glass slides. However, combination of multiple biomarkers from a wide variety of immunohistochemically stained slides is a tedious process in traditional histopathology due to the switching of glass slides and re-identification of regions of interest by pathologists. Digital pathology now allows us to apply image registration algorithms to digitized whole-slides to align the differing immunohistochemical stains automatically. However, registration algorithms need to be robust to changes in color due to differing stains and severe changes in tissue content between slides. In this work we developed a robust registration methodology to allow for fast coarse alignment of multiple immunohistochemical stains to the base hematyoxylin and eosin stained image. We applied HSD color model conversion to obtain a less stain color dependent representation of the whole-slide images. Subsequently, optical density thresholding and connected component analysis were used to identify the relevant regions for registration. Template matching using normalized mutual information was applied to provide initial translation and rotation parameters, after which a cost function-driven affine registration was performed. The algorithm was validated using 40 slides from 10 prostate cancer patients, with landmark registration error as a metric. Median landmark registration error was around 180 microns, which indicates performance is adequate for practical application. None of the registrations failed, indicating the robustness of the algorithm.

  17. Helicobacter pylori detection in chronic gastritis: a comparison of staining methods

    International Nuclear Information System (INIS)

    Ahmad, F.; Khan, I.

    2011-01-01

    Background: Helicobacter pylori is an important cause of chronic gastritis, gastric ulceration and gastric malignancies as gastric carcinoma and MALT lymphoma. Its definitive diagnosis is based on histopathology. Routine H and E stain is not very effective in its detection, immune-stains and fluorescent stains are costly. Need for simple cheap and sensitive stain has always been a topic of hot debate and extensive research. Method: paraffin embedded blocks of all adult patients diagnosed as chronic gastritis/gastric ulceration with no accompanying gastric pathology as hypertrophic gastropathys, and neoplasias were taken into study. Three sections of 4 micron were cut and stained with routine H and E, Giemsa, and Cresyl fast violet. Results: Total number of patients was 50. Out of these 37 (74%) were males and 13 (26%) were females. Mean age of the patients was 50.4 years. Thirty-four percent (34%) were positive in normal H and E stain, 68% were positive in Giemsa and 76% were positive in Cresyl fast violet. Conclusion: Cresyl fast violet is a good stain for diagnosis of H. pylori gastritis. (author)

  18. Quantitative magnetic resonance imaging (MRI) evaluation of cartilage repair after microfracture treatment for full-thickness cartilage defect models in rabbit knee joints: correlations with histological findings

    International Nuclear Information System (INIS)

    Tao, Hongyue; Feng, Xiaoyuan; Chen, Shuang; Li, Hong; Hua, Yinghui; Chen, Zhongqing

    2015-01-01

    To evaluate repair tissue (RT) after microfracture treatment for full-thickness cartilage defect models using quantitative MRI and investigate the correlations between MRI and histological findings. The animal experiment was approved by the Animal Care and Use Committee of our college. Thirty-six full-thickness cartilage defect models in rabbit knee joints were assigned to the microfracture or joint debridement group (as control). Each group consisted of 3-week, 5-week, and 7-week subgroups. MR imaging, including a three-dimensional double-echo steady-state sequence (3D-DESS), and T2 mapping were performed at 3, 5, and 7 weeks postoperatively. The thickness and T2 indices of RT were calculated. After MRI scans at each time point, operation sites were removed to make hematoxylin-eosin (H and E)-stained sections. Histological results were evaluated using the modified O'Driscoll score system. Comparisons were made between the two groups with respect to the MRI and histological findings, and correlation analysis was performed within each group. The thickness index and histological O'Driscoll score of RT in the two groups increased over time, while the T2 index decreased. The thickness index and histological O'Driscoll score of the microfracture group were higher than in the joint debridement group at each time point. The T2 index of the microfracture group was lower than in the joint debridement group at 3 weeks (P = 0.006), while it was higher than in the joint debridement group at 5 and 7 weeks (P = 0.025 and 0.025). The thickness index was positively correlated with the histological O'Driscoll score in both groups (microfracture: r s = 0.745, P s = 0.680, P = 0.002). The T2 index was negatively correlated with the histological O'Driscoll score in both groups (microfracture: r s = -0.715, P = 0.002; joint debridement: r s = -0.826, P < 0.001). Significant improvement over time after microfracture can be expected on the basis of the quantitative MRI finding and

  19. Quantitative magnetic resonance imaging (MRI) evaluation of cartilage repair after microfracture treatment for full-thickness cartilage defect models in rabbit knee joints: correlations with histological findings

    Energy Technology Data Exchange (ETDEWEB)

    Tao, Hongyue; Feng, Xiaoyuan; Chen, Shuang [Fudan University, Department of Radiology, Huashan Hospital, Shanghai (China); Li, Hong; Hua, Yinghui [Fudan University, Department of Sports Medicine, Huashan Hospital, Shanghai (China); Chen, Zhongqing [Fudan University, Department of Pathology, Huashan Hospital, Shanghai (China)

    2014-11-26

    To evaluate repair tissue (RT) after microfracture treatment for full-thickness cartilage defect models using quantitative MRI and investigate the correlations between MRI and histological findings. The animal experiment was approved by the Animal Care and Use Committee of our college. Thirty-six full-thickness cartilage defect models in rabbit knee joints were assigned to the microfracture or joint debridement group (as control). Each group consisted of 3-week, 5-week, and 7-week subgroups. MR imaging, including a three-dimensional double-echo steady-state sequence (3D-DESS), and T2 mapping were performed at 3, 5, and 7 weeks postoperatively. The thickness and T2 indices of RT were calculated. After MRI scans at each time point, operation sites were removed to make hematoxylin-eosin (H and E)-stained sections. Histological results were evaluated using the modified O'Driscoll score system. Comparisons were made between the two groups with respect to the MRI and histological findings, and correlation analysis was performed within each group. The thickness index and histological O'Driscoll score of RT in the two groups increased over time, while the T2 index decreased. The thickness index and histological O'Driscoll score of the microfracture group were higher than in the joint debridement group at each time point. The T2 index of the microfracture group was lower than in the joint debridement group at 3 weeks (P = 0.006), while it was higher than in the joint debridement group at 5 and 7 weeks (P = 0.025 and 0.025). The thickness index was positively correlated with the histological O'Driscoll score in both groups (microfracture: r{sub s} = 0.745, P < 0.001; joint debridement: r{sub s} = 0.680, P = 0.002). The T2 index was negatively correlated with the histological O'Driscoll score in both groups (microfracture: r{sub s} = -0.715, P = 0.002; joint debridement: r{sub s} = -0.826, P < 0.001). Significant improvement over time after

  20. Diagnostic value of HMB-45 and anti-Melan A staining of sentinel lymph nodes with isolated positive cells.

    Science.gov (United States)

    Mahmood, Muhammad N; Lee, Min W; Linden, Michael D; Nathanson, S D; Hornyak, Thomas J; Zarbo, Richard J

    2002-12-01

    Numerous immunohistochemical stains have been employed to detect metastatic melanoma in sentinel lymph node (SLN) biopsies. HMB-45 is considered by some as a specific tool to detect early metastatic melanoma (1). Occasionally, one or two isolated HMB-45-positive cells may cause complications in diagnostic interpretation. The goal of this study was to evaluate the reliability of HMB-45 staining of SLNs with sparse isolated positive cells and to compare its staining with anti-Melan A antibody. HMB-45 and anti-Melan A antibody immunostaining was performed on (Group A) 15 histologically negative SLNs excised from patients with malignant melanoma (MM) and on (Group B) 15 histologically negative SLNs excised from patients with breast carcinoma (BC). None of the patients had clinical evidence of systemic metastasis at the time of SLN biopsy. Five cutaneous biopsies with changes of postinflammatory hyperpigmentation (PIHP) were also stained with both antibodies. HMB-45 staining was repeated in all Group B SLNs after blocking endogenous biotins. Electron-microscopic studies were performed on all cases of PIHP. Isolated HMB-45-stained cells were present in 6 of 15 SLNs removed for MM; 8 of 15 for BC; and 3 of 5 cutaneous biopsies of PIHP. HMB-45 reactivity persisted after blocking endogenous biotins in 6 of 8 positive SLNs from Group B. Anti-Melan A antibody was negative in all SLNs of group A and B and in dermal melanophages of all five cases of PIHP. HMB-45 positivity was demonstrated in histologically negative SLNs and cutaneous biopsies, especially in the milieu of aggregated melanophages. Phagocytosis of premelanosomes by macrophages in the draining lymph nodes may account for isolated cell positivity and can hinder correct diagnostic interpretation. HMB-45 may not be a reliable marker for the detection of micro-metastasis of MM and requires correlation with other immunohistochemical markers, such as anti-Melan A antibody, to enhance specificity.

  1. Naturally acquired bovine besnoitiosis: histological and immunohistochemical findings in acute, subacute, and chronic disease.

    Science.gov (United States)

    Langenmayer, M C; Gollnick, N S; Majzoub-Altweck, M; Scharr, J C; Schares, G; Hermanns, W

    2015-05-01

    The pathogenesis of bovine besnoitiosis, a disease of increasing concern within Europe, is still incompletely understood. In this study, disease progression after natural infection with the causative apicomplexan Besnoitia besnoiti was monitored in histological skin sections of 5 individual female cattle over time. High-frequency skin sampling of 2 cattle with mild and 2 with severe acute, subacute, and chronic besnoitiosis, as well as from 1 animal during subclinical disease, enabled documentation from the beginning of the disease. Skin sections were stained with hematoxylin and eosin and Giemsa, periodic acid-Schiff reaction, and anti-Besnoitia immunohistochemistry. In all 4 clinically affected animals, tachyzoite-like endozoites could be detected for the first time by immunohistochemistry, and tissue cyst evolution was monitored. Besnoitiosis-associated lesions were not detected in the animal showing the subclinical course. Because of the inconsistency of the nomenclature of Besnoitia tissue cyst layers in the literature, a new nomenclature for B. besnoiti cyst wall layers is proposed: tissue cysts consist of a hypertrophied host cell with enlarged nuclei, an intracytoplasmic parasitophorous vacuole with bradyzoites, a sometimes vacuolated inner cyst wall, and an outer cyst wall in more developed cysts. Inner and outer cyst walls can be readily distinguished by using special stains. In 1 animal, extracystic B. besnoiti zoites were immunohistochemically detected during the chronic stage. At necropsy, the 2 severely affected cows displayed large numbers of B. besnoiti cysts in a variety of tissues, including the corium of the claws, contributing mainly to the development of chronic laminitis in these 2 cases. © The Author(s) 2014.

  2. Quantification of tumor morphology via 3D histology: application to oral cavity cancers

    Science.gov (United States)

    Doyle, Scott; Brandwein-Gensler, Margaret; Tomaszewski, John

    2016-03-01

    Traditional histopathology quantifies disease through the study of glass slides, i.e. two-dimensional samples that are representative of the overall process. We hypothesize that 3D reconstruction can enhance our understanding of histopathologic interpretations. To test this hypothesis, we perform a pilot study of the risk model for oral cavity cancer (OCC), which stratifies patients into low-, intermediate-, and high-risk for locoregional disease-free survival. Classification is based on study of hematoxylin and eosin (H and E) stained tissues sampled from the resection specimens. In this model, the Worst Pattern of Invasion (WPOI) is assessed, representing specific architectural features at the interface between cancer and non-cancer tissue. Currently, assessment of WPOI is based on 2D sections of tissue, representing complex 3D structures of tumor growth. We believe that by reconstructing a 3D model of tumor growth and quantifying the tumor-host interface, we can obtain important diagnostic information that is difficult to assess in 2D. Therefore, we introduce a pilot study framework for visualizing tissue architecture and morphology in 3D from serial sections of histopathology. This framework can be used to enhance predictive models for diseases where severity is determined by 3D biological structure. In this work we utilize serial H and E-stained OCC resections obtained from 7 patients exhibiting WPOI-3 (low risk of recurrence) through WPOI-5 (high risk of recurrence). A supervised classifier automatically generates a map of tumor regions on each slide, which are then co-registered using an elastic deformation algorithm. A smooth 3D model of the tumor region is generated from the registered maps, which is suitable for quantitative tumor interface morphology feature extraction. We report our preliminary models created with this system and suggest further enhancements to traditional histology scoring mechanisms that take spatial architecture into consideration.

  3. Histological study of white rhinoceros integument.

    Directory of Open Access Journals (Sweden)

    Jeffrey H Plochocki

    Full Text Available In this study, we report findings from a microscopic analysis of the white rhinoceros (Ceratotherium simum integumentary ultrastructure. Skin samples from the cheek, shoulder, flank and rump were taken from a 46-year-old female southern white rhinoceros and examined using H&E and elastic histological stains. The epidermis was thickest in the flank (1.003 mm followed by the rump, cheek and shoulder. The stratum corneum comprised more than half the epidermal thickness. Numerous melanin granules were found in the basal and spinosum layers. The epidermal-dermal junction was characterized by abundant papillary folds increasing surface contact between integument layers. Most of the dermal thickness consisted of organized collagen bundles with scattered elastic fibers. Collagen fiber bundles were thickest in the flank (210.9 μm followed by shoulder, rump and cheek. Simple coiled sweat glands were present in the dermis, but hair and sebaceous glands were absent. Together, these data suggest the white rhinoceros has a unique integumentary system among large terrestrial herbivores.

  4. Histological study of white rhinoceros integument.

    Science.gov (United States)

    Plochocki, Jeffrey H; Ruiz, Saul; Rodriguez-Sosa, José R; Hall, Margaret I

    2017-01-01

    In this study, we report findings from a microscopic analysis of the white rhinoceros (Ceratotherium simum) integumentary ultrastructure. Skin samples from the cheek, shoulder, flank and rump were taken from a 46-year-old female southern white rhinoceros and examined using H&E and elastic histological stains. The epidermis was thickest in the flank (1.003 mm) followed by the rump, cheek and shoulder. The stratum corneum comprised more than half the epidermal thickness. Numerous melanin granules were found in the basal and spinosum layers. The epidermal-dermal junction was characterized by abundant papillary folds increasing surface contact between integument layers. Most of the dermal thickness consisted of organized collagen bundles with scattered elastic fibers. Collagen fiber bundles were thickest in the flank (210.9 μm) followed by shoulder, rump and cheek. Simple coiled sweat glands were present in the dermis, but hair and sebaceous glands were absent. Together, these data suggest the white rhinoceros has a unique integumentary system among large terrestrial herbivores.

  5. A topical fluorescent analogue for virtual hematoxylin and eosin histology in point-of-care ex vivo microscopy

    Science.gov (United States)

    Elfer, Katherine; Sholl, Andrew; Miller, Christopher; Brown, J. Quincy

    2015-07-01

    Histological assessment of freshly removed tissue specimens requires accurate and fast analysis in clinical procedures such as diagnostic biopsy and surgical tumor resection. Current histological assessment methods are either time-consuming or damage the tissue beyond the ability to re-analyze post-procedure. We demonstrate a novel dual-stain fluorescent analogue to brightfield Hematoxylin and Eosin for in-procedure histopathology that is both time-efficient and preserves the analyzed tissue for later analysis. H&E-like images are created from the combination of DRAQ5 and Eosin applied to human prostate tissue and animal muscle tissue under confocal microscopy. D&E images are pseduocolored to match H&E coloring, showing near-identical features to brightfield H&E of the same tissue. The histological accuracy, short staining time, and tissue preservation aspects of this dual-stain technique demonstrates its potential to be adopted for use in point-of-care pathology.

  6. Fluoro jade-C staining in the assessment of brain injury after deep hypothermia circulatory arrest.

    Science.gov (United States)

    Wang, Ren; Ma, Wei-Guo; Gao, Guo-Dong; Mao, Qun-Xia; Zheng, Jun; Sun, Li-Zhong; Liu, Ying-Long

    2011-02-04

    To evaluate the efficacy of Fluoro Jade-C staining (FJC) in the assessment of brain injury after deep hypothermia circulatory arrest (DHCA). Six healthy adult miniature male pigs underwent DHCA, the rectal temperature was down to 18°C, circulation was stopped , circulatory arrest was maintained for 60 minutes. On postoperative day 1, perfusion-fixation was performed on brain tissue. Cerebral cortex, hippocampus, cerebellum were taken for sampling. FJC, hematoxylin-eosin staining (HE), nissl staining (NISSL), terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) were performed to detect the histological and pathological changes. Histological scores of all slices were ranked. Comparison between the FJC and other techniques was done by analysis of variance (ANOVA) according to histological scores. All animals survived the operation. On the cerebral cortex, in comparison of FJC between HE, NISSL and TUNEL, the p value was 0.90, 0.40, 0.16 respectively (p>0.05). On the hippocampus, the comparison of FJC with HE, NISSL and TUNEL had a p value of 0.12, 0.23, 0.62 respectively (p>0.05). On the cerebellum, in comparing FJC with HE, NISSL and TUNEL, the p value was 0.96, 0.77, 0.96 respectively (p>0.05). On representative regions, the results of FJC were in accordance with that of TUNEL, NISSL and HE. Furthermore, ascertainment of brain injury is easier with FJC. FJC is a reliable and convenient method to assess brain injury after DHCA. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. A Simple and Efficient Method for Preparing Cell Slides and Staining without Using Cytocentrifuge and Cytoclips

    Directory of Open Access Journals (Sweden)

    Xiaotang Hu

    2015-01-01

    Full Text Available Cell staining is a necessary and useful technique for visualizing cell morphology and structure under a microscope. This technique has been used in many areas such as cytology, hematology, oncology, histology, virology, serology, microbiology, cell biology, and immunochemistry. One of the key pieces of equipment for preparing a slide for cell staining is cytology centrifuge (cytocentrifuge such as cytospin. However, many small labs do not have this expensive equipment and its accessory, cytoclips (also expensive relatively, which makes them difficult to study cell cytology. Here we present an alternative method for preparing a slide and cell staining in the absence of a cytocentrifuge (and cytoclips. This method is based on the principle that a regular cell centrifuge can be used to concentrate cells harvested from cell culture and then deposit the concentrated cell suspension to a slide evenly by using a cell spreader, followed by cell staining. The method presented is simple, rapid, economic, and efficient. This method may also avoid a possible change in cell morphology induced by cytocentrifuge.

  8. A rapid method combining Golgi and Nissl staining to study neuronal morphology and cytoarchitecture.

    Science.gov (United States)

    Pilati, Nadia; Barker, Matthew; Panteleimonitis, Sofoklis; Donga, Revers; Hamann, Martine

    2008-06-01

    The Golgi silver impregnation technique gives detailed information on neuronal morphology of the few neurons it labels, whereas the majority remain unstained. In contrast, the Nissl staining technique allows for consistent labeling of the whole neuronal population but gives very limited information on neuronal morphology. Most studies characterizing neuronal cell types in the context of their distribution within the tissue slice tend to use the Golgi silver impregnation technique for neuronal morphology followed by deimpregnation as a prerequisite for showing that neuron's histological location by subsequent Nissl staining. Here, we describe a rapid method combining Golgi silver impregnation with cresyl violet staining that provides a useful and simple approach to combining cellular morphology with cytoarchitecture without the need for deimpregnating the tissue. Our method allowed us to identify neurons of the facial nucleus and the supratrigeminal nucleus, as well as assessing cellular distribution within layers of the dorsal cochlear nucleus. With this method, we also have been able to directly compare morphological characteristics of neuronal somata at the dorsal cochlear nucleus when labeled with cresyl violet with those obtained with the Golgi method, and we found that cresyl violet-labeled cell bodies appear smaller at high cellular densities. Our observation suggests that cresyl violet staining is inadequate to quantify differences in soma sizes.

  9. Methylene Blue-Aided In Vivo Staining of Central Airways during Flexible Bronchoscopy

    Directory of Open Access Journals (Sweden)

    Sabine Zirlik

    2012-01-01

    Full Text Available Background. The early diagnosis of malignant and premalignant changes of the bronchial mucosa remains a major challenge during bronchoscopy. Intravital staining techniques are not new. Previous small case series suggested that analysis of the bronchial mucosal surface using chromoendoscopy allows a prediction between neoplastic and nonneoplastic lesions. Objectives. The aim of the present study was to evaluate chromobronchoscopy as a method to identify malignant and premalignant lesions in the central airways in a prospective manner. Methods. In 26 patients we performed chromoendoscopy with 0.1% methylene blue during ongoing flexible white light bronchoscopy. Circumscribed lesions in central airways were further analyzed by biopsies and histopathologic examination. Results. In the majority of cases neither flat nor polypoid lesions in the central airways were stained by methylene blue. In particular, exophytic growth of lung cancer did not show any specific pattern in chromobronchoscopy. However, a specific dye staining was detected in one case where exophytic growth of metastatic colorectal cancer was present in the right upper lobe. In two other cases, a circumscribed staining was noted in unsuspicious mucosa. But histology revealed inflammation only. Conclusions. In contrast to previous studies, the present findings clearly indicate that chromobronchoscopy is not useful for early detection of malignant or premalignant lesions of the central airways.

  10. Block-Centric Visualization Of Histological Whole Slide Images With Application To Revealing Growth-Patterns Of Early Colorectal Adenomas And Aberrant Crypt Foci

    Directory of Open Access Journals (Sweden)

    Norman Zerbe

    2016-06-01

    The presented methods provide an efficient means to retrieve correspondences and additional spatial information from serial sections of histological slides. They also show good applicability for specimen from different origin. Alignment methods can be applied to generate block-centric visualizations such as parallel and transparent viewing of multiple stains. Moreover, the generated stack videos and 3D models demonstrate the very good accuracy of section alignment even in large series. The visualizations enable pathologists and researchers to grasp the 3D structural relationships in the tissue at a glance, providing an excellent tool to communicate more complex histomorphological findings. Interestingly, we see two kinds of tubular adenomas, which could imply multiple ways to tubular adenoma formation in FAP-patients, possibly akin to the recent observations in mice [3].

  11. Hierarchical patch-based co-registration of differently stained histopathology slides

    Science.gov (United States)

    Yigitsoy, Mehmet; Schmidt, Günter

    2017-03-01

    Over the past decades, digital pathology has emerged as an alternative way of looking at the tissue at subcellular level. It enables multiplexed analysis of different cell types at micron level. Information about cell types can be extracted by staining sections of a tissue block using different markers. However, robust fusion of structural and functional information from different stains is necessary for reproducible multiplexed analysis. Such a fusion can be obtained via image co-registration by establishing spatial correspondences between tissue sections. Spatial correspondences can then be used to transfer various statistics about cell types between sections. However, the multi-modal nature of images and sparse distribution of interesting cell types pose several challenges for the registration of differently stained tissue sections. In this work, we propose a co-registration framework that efficiently addresses such challenges. We present a hierarchical patch-based registration of intensity normalized tissue sections. Preliminary experiments demonstrate the potential of the proposed technique for the fusion of multi-modal information from differently stained digital histopathology sections.

  12. Immunogold staining procedure for the localisation of regulatory peptides.

    Science.gov (United States)

    Varndell, I M; Tapia, F J; Probert, L; Buchan, A M; Gu, J; De Mey, J; Bloom, S R; Polak, J M

    1982-01-01

    The use of protein A- and IgG-conjugated colloidal gold staining methods for the immuno-localisation of peptide hormones and neurotransmitters at light- and electron microscope level are described and discussed. Bright-field and dark-ground illumination modes have been used to visualise the gold-labelled antigenic sites at the light microscope level. Immunogold staining procedures at the ultrastructural level using region-specific antisera have been adopted to localise specific molecular forms of peptides including gastrin (G17 and G34), glucagon and pro-glucagon, insulin and pro-insulin, in normal tissue and in tumours of the gastroenteropancreatic system. Similar methods have been used to demonstrate the heterogeneity of p-type nerves in the enteric nervous system. Vasoactive intestinal polypeptide (VIP) has been localised to granular sites (mean +/- S.D. granule diameter = 98 +/- 19 nm) in nerve terminals of the enteric plexuses and in tumour cells of diarrhoeogenic VIP-producing neoplasias (mean +/- S.D. granule diameter = 126 +/- 37 nm) using immunogold procedures applied to ultraviolet-cured ultrathin sections. Co-localisation of amines and peptides in carotid body type I cells and in chromaffin cells of normal adrenal medulla and phaeochromocytomas has also been demonstrated. Advantages of the immunogold procedures over alternative immunocytochemical techniques are discussed.

  13. Stained glasses under the nuclear microprobe: A window into history

    Energy Technology Data Exchange (ETDEWEB)

    Vilarigues, M. [Dep. de Conservacao e Restauro and R and D Unit Vidro e da Ceramica Para as Artes, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal)], E-mail: mgv@fct.unl.pt; Fernandes, P. [Dep. de Conservacao e Restauro and R and D Unit Vidro e da Ceramica Para as Artes, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Alves, L.C.; Silva, R.C. da [Dep. Fisica, LFI, ITN, E.N.10, 2686-953 Sacavem (Portugal)

    2009-06-15

    Stained glass fragments from the 15th, 16th and 20th centuries, belonging to Mosteiro de Santa Maria da Vitoria, Batalha (Portugal), were characterised non-destructively in a nuclear microprobe. The work aimed at finding the composition of the glasses and glass paintings and relating these with the corresponding production periods. The elemental compositions of the glass fragments were obtained by means of scanning micro-beam Particle Induced X-ray Emission ({mu}-PIXE) spectrometry in selected cross-sections. These were complemented by micro X-Ray fluorescence spectrometry. Characterisation of colour was performed by optical absorption spectroscopy in the UV-vis range, while the corrosion products were identified by optical microscopy and {mu}-FTIR (Fourier Transform Infra Red) spectroscopy in combination with the data generated by {mu}-PIXE. Nuclear microprobe analysis allowed unveiling the compositions and structures, in particular of glass paintings and corrosion products. While it is not surprising that Fe, Cu and Pb were the main elements identified in the grisaille paintings of all studied periods, as well as Ag and Cu found in the glasses decorated with yellow silver painting, their distribution gave important clues on the materials and techniques used to manufacture these stained glasses. Furthermore, it allowed establishing a definite relation between the compositions found and the periods of production, with the added bonus of correctly reassigning the manufacturing period of some samples.

  14. Reordering Histology to Enhance Engagement

    Science.gov (United States)

    Amerongen, Helen

    2011-01-01

    In redesigning the preclinical curriculum and shifting from a discipline-based approach to an organ system-based approach, faculty at the University of Arizona College of Medicine in Tucson took the opportunity to restructure the sequence of introductory histology content to make it more engaging and relevant. In this article, the author describes…

  15. Protective effect of ellagic acid on healing alveolar bone after tooth extraction in rat--a histological and immunohistochemical study.

    Science.gov (United States)

    Al-Obaidi, Mazen M Jamil; Al-Bayaty, Fouad Hussain; Al Batran, Rami; Hassandarvish, Pouya; Rouhollahi, Elham

    2014-09-01

    This study has attempted to evaluate the effects of ellagic acid (EA) on alveolar bone healing after tooth extraction in rats. Twenty-four Sprague Dawley (SD) male rats (200-250g) were selected and were anaesthetised for the extraction of upper left incisor. Then, the rats were divided into two groups, comprising 12 rats each; the first group has been considered as a control group and was given only normal saline, whereas, the second group (treated group) was intragastrically administrated with EA daily once, for 28 days. Then three rats from each group had been selected on 7th, 14th, 21st, and 28th days to dissect their maxilla tissue either for histological observation and homogenisation purposes. The tissues fixed, decalcified and embedded in paraffin. Serial sections of 5μm thickness were prepared and stained with haematoxylin and eosin (H&E) for the histological study. Similar sections were taken for immunohistochemical analysis to assess osteocalcin (OSC) and osteopontin (OPN). Furthermore, Malondialdehyde (MDA) and superoxide dismutase (SOD) were measured in homogenated gingival maxilla tissue of rat by commercial kit. Based on the histological analysis we have identified that, EA treatment has induced earlier trabecular bone deposition in the treated group, resulting in more organised bone matrix on the 14th, 21st, and 28th days after tooth extraction, as against the control group. In comparison to control group, the positive labelling of OSC and OPN of the treated group have been highly expressed in the alveolar socket on 14th, and 21st days, which has indicated a the possibility of formation of new bone trabeculae at the beginning of the mineralisation process, after tooth extraction. In the EA treatment group, lipid per-oxidation (MDA) was significantly decreased (Phealing process in teeth socket of rats. Furthermore, the EA treated group showed a stronger positive immunolabelling for OSC and OPN, when compared with the control group. Copyright © 2014

  16. WITHDRAWN: Amnioinfusion for meconium-stained liquor in labour.

    Science.gov (United States)

    Hofmeyr, G Justus

    2009-01-21

    Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. It is also thought to dilute meconium when present in the amniotic fluid and so reduce the risk of meconium aspiration. However, it may be that the mechanism of effect is that it corrects oligohydramnios (reduced amniotic fluid), for which thick meconium staining is a marker. The objective of this review was to assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. The Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (Issue 3, 2001) were searched. Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium-staining of the amniotic fluid. Eligibility and trial quality were assessed by one reviewer. Twelve studies, most involving small numbers of participants, were included. Under standard perinatal surveillance, amnioinfusion was associated with a reduction in the following: heavy meconium staining of the liquor (relative risk 0.03, 95% confidence interval 0.01 to 0.15); variable fetal heart rate deceleration (relative risk 0.65, 95% confidence interval 0.49 to 0.88); and reduced caesarean section overall (relative risk 0.82, 95% confidence interval 0.69 to 1.97). No perinatal deaths were reported. Under limited perinatal surveillance, amnioinfusion was associated with a reduction in the following: meconium aspiration syndrome (relative risk 0.24, 95% confidence interval 0.12 to 0.48); neonatal hypoxic ischaemic encephalopathy (relative risk 0.07, 95% confidence interval 0.01 to 0.56) and neonatal ventilation or intensive care unit admission (relative risk 0.56, 95% confidence interval 0.39 to 0.79); there was a trend towards reduced perinatal mortality (relative risk 0.34, 95% confidence interval 0.11 to 1.06). Amnioinfusion is associated with improvements in perinatal outcome

  17. High-Throughput Method of Whole-Brain Sectioning, Using the Tape-Transfer Technique.

    Science.gov (United States)

    Pinskiy, Vadim; Jones, Jamie; Tolpygo, Alexander S; Franciotti, Neil; Weber, Kevin; Mitra, Partha P

    2015-01-01

    Cryostat sectioning is a popular but labor-intensive method for preparing histological brain sections. We have developed a modification of the commercially available CryoJane tape collection method that significantly improves the ease of collection and the final quality of the tissue sections. The key modification involves an array of UVLEDs to achieve uniform polymerization of the glass slide and robust adhesion between the section and slide. This report presents system components and detailed procedural steps, and provides examples of end results; that is, 20 μm mouse brain sections that have been successfully processed for routine Nissl, myelin staining, DAB histochemistry, and fluorescence. The method is also suitable for larger brains, such as rat and monkey.

  18. Port wine stain on a child's face (image)

    Science.gov (United States)

    Port wine stains are always present at birth. In an infant, they are flat, pink, vascular lesions. Common locations ... may be present anywhere on the body. Port wine stains may appear in association with other syndromes.

  19. Crosslinking and corneal cryotherapy in acanthamoeba keratitis -- a histological study.

    Science.gov (United States)

    Hager, Tobias; Hasenfus, A; Stachon, T; Seitz, B; Szentmáry, N

    2016-01-01

    Acanthamoeba keratitis is rare, but difficult to treat. Penetrating keratoplasty is performed in therapy-resistant cases. Nevertheless, subsequent recurrences occur in 40 % of the cases. In addition to triple-topical therapy (polyhexamid, propamidinisoethionat, neomycin), treatment alternatives are corneal cryotherapy and/or crosslinking (CXL). The aim of our present histological study was to analyze the persistence of acanthamoebatrophozoites and cysts, the persistence of bacteria, and activation of keratocytes in corneas of acanthamoeba keratitis patients following corneal cryotherapy and/or CXL. We analyzed histologically corneal buttons (from penetrating keratoplasties) of nine patients with acanthamoeba keratitis, following corneal cryotherapy (two patients) or a combination of crosslinking and corneal cryotherapy (seven patients), using haematoxilin–eosin, periodic acid Schiff (PAS), Gram and alpha-smooth muscle actin (alpha-SMA) stainings. Acanthamoeba trophozoites persisted in three corneas after cryotherapy and CXL. Cysts persisted in one of two corneas following corneal cryotherapy and in six of seven corneas after a combination of CXL and cryotherapy. One cornea showed positive Gram staining, but there were no alpha-SMA positive keratocytes in any of the corneas. Crosslinking and corneal cryotherapy have only limited impact on killing of acanthamoeba trophozoites, cysts, or bacteria. Corneal cryotherapy and CXL did not stimulate myofibroblastic transformation of keratocytes.

  20. Carbonic anhydrase XII expression is associated with histologic grade of cervical cancer and superior radiotherapy outcome

    International Nuclear Information System (INIS)

    Yoo, Chong Woo; Nam, Byung-Ho; Kim, Joo-Young; Shin, Hye-Jin; Lim, Hyunsun; Lee, Sun; Lee, Su-Kyoung; Lim, Myong-Cheol; Song, Yong-Jung

    2010-01-01

    To investigate whether expression of carbonic anhydrase XII (CA12) is associated with histologic grade of the tumors and radiotherapy outcomes of the patients with invasive cervical cancer. CA12 expression was examined by immunohistochemical stains in cervical cancer tissues from 183 radiotherapy patients. Histological grading was classified as well (WD), moderately (MD) or poorly differentiated (PD). Oligonucleotide microarray experiment was performed using seven cervical cancer samples to examine differentially expressed genes between WD and PD cervical cancers. The association between CA12 and histological grade was analyzed by chi-square test. CA12 and histological grades were analyzed individually and as combined CA12 and histologic grade categories for effects on survival outcome. Immunohistochemical expression of CA12 was highly associated with the histologic grade of cervical cancer. Lack of CA12 expression was associated with PD histology, with an odds ratio of 3.9 (P = 0.01). Microarray analysis showed a fourfold reduction in CA12 gene expression in PD tumors. CA12 expression was marginally associated with superior disease-free survival. Application of the new combined categories resulted in further discrimination of the prognosis of patients with moderate and poorly differentiated tumor grade. Our study indicates that CA12 may be used as a novel prognostic marker in combination with histologic grade of the tumors

  1. Utility of Modified Ultrafast Papanicolaou Stain in Cytological Diagnosis.

    Science.gov (United States)

    Sinkar, Prachi; Arakeri, Surekha Ulhas

    2017-03-01

    Need for minimal turnaround time for assessing Fine Needle Aspiration Cytology (FNAC) has encouraged innovations in staining techniques that require lesser staining time with unequivocal cell morphology. The standard protocol for conventional Papanicolaou (PAP) stain requires about 40 minutes. To overcome this, Ultrafast Papanicolaou (UFP) stain was introduced which reduces staining time to 90 seconds and also enhances the quality. However, reagents required for this were not easily available hence, Modified Ultrafast Papanicolaou (MUFP) stain was introduced subsequently. To assess the efficacy of MUFP staining by comparing the quality of MUFP stain with conventional PAP stain. FNAC procedure was performed by using 10 ml disposable syringe and 22-23 G needle. Total 131 FNAC cases were studied which were lymph node (30), thyroid (38), breast (22), skin and soft tissue (24), salivary gland (11) and visceral organs (6). Two smears were prepared and stained by MUFP and conventional PAP stain. Scores were given on four parameters: background of smears, overall staining pattern, cell morphology and nuclear staining. Quality Index (QI) was calculated from ratio of total score achieved to maximum score possible. Statistical analysis using chi square test was applied to each of the four parameters before obtaining the QI in both stains. Students t-test was applied to evaluate the efficacy of MUFP in comparison with conventional PAP stain. The QI of MUFP for thyroid, breast, lymph node, skin and soft tissue, salivary gland and visceral organs was 0.89, 0.85, 0.89, 0.83, 0.92, and 0.78 respectively. Compared to conventional PAP stain QI of MUFP smears was better in all except visceral organ cases and was statistically significant. MUFP showed clear red blood cell background, transparent cytoplasm and crisp nuclear features. MUFP is fast, reliable and can be done with locally available reagents with unequivocal morphology which is the need of the hour for a cytopathology set-up.

  2. Discriminative staining methods for the nervous system: luxol fast blue--periodic acid-Schiff--hematoxylin triple stain and subsidiary staining methods.

    Science.gov (United States)

    Goto, N

    1987-09-01

    This paper describes a new series of staining methods which can discriminatively demonstrate every structure of the nervous system, including axons and capillaries, in animal and human materials. Methods described in this paper consist of one primary stain, luxol fast blue-periodic acid Schiff-hematoxylin (LPH) and six different subsidiary staining methods. The LPH triple stain can precisely differentiate the following structures: neurons (Nissl bodies, cytoplasm, nuclear membrane and nucleolus), various kinds of nuclei (glia, ependyma, endothelium, leucocyte, connective tissue, etc.), myelin sheaths, neuronal processes (axons and dendrites), reacted glial cell bodies (protoplasmic astrocytes, foamy cells, etc.), blood vessels (arteries, veins and capillaries), meninges, intervening connective tissue, erythrocytes, lipofuscin granules, amyloid bodies, and others. Subsidiary staining methods are also described briefly. Applications are discussed in the context of staining technology and neuromorphological research.

  3. Vitamin D3 May Ameliorate the Ketoconazole Induced Adrenal Injury: Histological and Immunohistochemical Studies on Albino Rats

    International Nuclear Information System (INIS)

    Khalil, Mahmoud Salah

    2015-01-01

    Ketoconazole (KZ) is used widely for treating the superficial, systemic fungal activities and hyperandrogenemic states. Its uses are limited by its deleterious effect on histological structure and function of the adrenal cortex. This study investigates whether vitamin D3 supplement can ameliorate the morphological changes induced by KZ. Thirty four adult male albino rats were randomized into control group (Group I) which was subdivided into: control 1 (n=7) and control 2 (n=7): In control 1, rats were intraperitoneal (I.P) injected once with 1 ml of polyethylene glycol-400 for 15 consecutive days and control 2 rats were injected I.P with (1 μg/kg) of vitamin D3 for the same period. Group II (n=10): rats were I.P injected with KZ (10 mg/100 g of body weight) once daily for 15 days; Group III (n=10): rats were I.P concomitantly injected with KZ and vitamin D3 similar doses to animals in groups II and control 2 respectively. Blood samples were collected to determine plasma ACTH, corticosterone and aldosterone levels. The right adrenal specimens sections were stained with Haematoxylin & Eosin and Masson Trichrome for histological studies and treated with Bax, Ubiquitin and vitamin D receptors for immunohistochemical studies. KZ induced adrenal cortical morphological changes in forms of disturbed adrenocorticocyte cytological architecture, nuclear changes, and intracellular lipid accumulation. KZ also increased adrenal Bax and Ub but decreased the vitamin D receptors immunopositive staining expression, in addition to increased plasma ACTH as well as decreased corticosterone and aldosterone levels. These changes were ameliorated by supplementing with vitamin D3

  4. CD3 immunohistochemical staining in diagnosis of lymphocytic colitis

    DEFF Research Database (Denmark)

    Fiehn, Anne-Marie Kanstrup; Engel, Ulla; Holck, Susanne

    2016-01-01

    and eosin (HE) stainings were available. At the second assessment, a supplementary CD3 immunohistochemical staining was also available. The aim was to evaluate whether a supplementary CD3 would increase the diagnostic agreement among pathologists, and whether a CD3 stain would change the diagnosis based...

  5. Evaluation of biosafe alternatives as xylene substitutes in hematoxylin and eosin staining procedure: A comparative pilot study

    Science.gov (United States)

    Sravya, Taneeru; Rao, Guttikonda Venkateswara; Kumari, Masabattula Geetha; Sagar, Yerraguntla Vidya; Sivaranjani, Yeluri; Sudheerkanth, Kondamarri

    2018-01-01

    Background: Xylene is synthetic hydrocarbon produced from coal tar known for its wide usage as universal solvent which has many hazardous effects. The aim of this study is to compare the efficacy of xylene-free hematoxylin and eosin (H and E) sections with conventional H and E sections. Materials and Methods: The study included ninety paraffin-embedded tissue blocks. Of these, sixty blocks were processed with sesame oil (xylene alternative) and thirty blocks with xylene. The study sample was divided into three groups. Sixty sections which are taken from sesame oil-processed blocks were stained with xylene-free H and E staining method. In xylene-free staining method, 95% diluted lemon water (Group A) and 1.7% dish washing solution (DWS, Group B) were used as deparaffinizing agents whereas the remaining 30 sections were processed with xylene and stained with conventional H and E staining method (Group C). Slides were scored for the following parameters: (i) nuclear staining (adequate = score 1, inadequate = score 0), (ii) cytoplasmic staining (adequate = score 1, inadequate = score 0), (iii) uniformity (present = score 1, absent = score 0), (iv) clarity (present = score 1, absent = score 0) and (v) intensity (present = score 1, absent = score 0). Score ≤2 was considered inadequate for diagnosis while scores 3–5 were considered adequate for diagnosis. Results: Adequate nuclear staining was noted in 90% of sections of Group A and 100% each in Group B and Group C (P 0.05); adequate uniformity of staining in 53.3% of sections of Group A, 70% in Group B and 83.3% in Group C (P 0.05) and adequate intensity of staining in 76.7% sections of Group A, 93.3% in Group B and 100% in Group C (P < 0.05). Group C sections stained adequate for diagnosis (93.3%) followed by Group B (88.7%) and Group A (78%; P < 0.05). Conclusion: Tissues processed with sesame oil and stained using 1.7% DWS were found to be effective alternative to xylene.

  6. Cell wall staining with Trypan Blue enables quantitative analysis of morphological changes in yeast cells

    Directory of Open Access Journals (Sweden)

    Johannes eLiesche

    2015-02-01

    Full Text Available Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays.

  7. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells.

    Science.gov (United States)

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays.

  8. Quantitative Measurements in the Human Hippocampus and Related Areas: Correspondence between Ex-Vivo MRI and Histological Preparations.

    Directory of Open Access Journals (Sweden)

    José Carlos Delgado-González

    Full Text Available The decrease of volume estimates in different structures of the medial temporal lobe related to memory correlate with the decline of cognitive functions in neurodegenerative diseases. This study presents data on the association between MRI quantitative parameters of medial temporal lobe structures and their quantitative estimate in microscopic examination. Twelve control cases had ex-vivo MRI, and thereafter, the temporal lobe of both hemispheres was sectioned from the pole as far as the level of the splenium of the corpus callosum. Nissl stain was used to establish anatomical boundaries between structures in the medial temporal lobe. The study included morphometrical and stereological estimates of the amygdaloid complex, hippocampus, and temporal horn of the lateral ventricle, as well as different regions of grey and white matter in the temporal lobe. Data showed a close association between morphometric MRI images values and those based on the histological determination of boundaries. Only values in perimeter and circularity of the piamater were different. This correspondence is also revealed by the stereological study, although irregular compartments resulted in a lesser agreement. Neither age ( 65 yr nor hemisphere had any effect. Our results indicate that ex-vivo MRI is highly associated with quantitative information gathered by histological examination, and these data could be used as structural MRI biomarker in neurodegenerative diseases.

  9. A histological and immunohistochemical study of the effects of N-acetyl cysteine on retinopathy of prematurity by modifying insulin-like growth factor-1.

    Science.gov (United States)

    El-Hadidy, A R; El-Mohandes, E M; Asker, S A; Ghonaim, F M

    2016-08-01

    Retinopathy of prematurity (ROP) is a vasoproliferative disorder that occurs in premature infants and may lead to permanent visual impairment. We investigated both the possible protective role of N-acetyl cysteine (NAC) for preventing ROP and the role of IGF-1 in the disorder. Forty-five newborn rats were divided into three groups. Group 1 was raised in room air as controls. Group 2 was exposed to 60% oxygen for 14 days after birth, then transferred to room air. Group 3 was exposed to the same conditions as group 2, but received intraperitoneal injections of NAC on postnatal days 7-17. After 35 days, both eyes of all rats were processed for histology. Some sections were stained with hematoxylin and eosin to assess structural changes and other sections were immunostained to determine the location of IGF-1. Frozen sections also were prepared and stained for adenosine triphosphatase to detect retinal blood vessels. Compared to the controls, more blood vessels, many of which were abnormal, and increased IGF-1 expression were observed in group 2. In group 3, abnormal blood vessels and IGF-1 expression were less evident. NAC appeared to be an effective vascular-protective agent for ROP by decreasing IGF-1 expression.

  10. Verification of the anatomy and newly discovered histology of the G-spot complex.

    Science.gov (United States)

    Ostrzenski, A; Krajewski, P; Ganjei-Azar, P; Wasiutynski, A J; Scheinberg, M N; Tarka, S; Fudalej, M

    2014-10-01

    To expand the anatomical investigations of the G-spot and to assess the G-spot's characteristic histological and immunohistochemical features. An observational study. International multicentre. Eight consecutive fresh human female cadavers. Anterior vaginal wall dissections were executed and G-spot microdissections were performed. All specimens were stained with haematoxylin and eosin (H&E). The tissues of two women were selected at random for immunohistochemical staining. The primary outcome measure was to document the anatomy of the G-spot. The secondary outcome measures were to identify the histology of the G-spot and to determine whether histological samples stained with H&E are sufficient to identify the G-spot. The anatomical existence of the G-spot was identified in all women and was in a diagonal plane. In seven (87.5%) and one (12.5%) of the women the G-spot complex was found on the left or right side, respectively. The G-spot was intimately fused with vessels, creating a complex. A large tangled vein-like vascular structure resembled an arteriovenous malformation and there were a few smaller feeding arteries. A band-like structure protruded from the tail of the G-spot. The size of the G-spot varied. Histologically, the G-spot was determined as a neurovascular complex structure. The neural component contained abundant peripheral nerve bundles and a nerve ganglion. The vascular component comprised large vein-like vessels and smaller feeding arteries. Circular and longitudinal muscles covered the G-complex. The anatomy of the G-spot complex was confirmed. The histology of the G-spot presents as neurovascular tissues with a nerve ganglion. H&E staining is sufficient for the identification of the G-spot complex. © 2014 Royal College of Obstetricians and Gynaecologists.

  11. Centrifuge-operated specimen staining method and apparatus

    Science.gov (United States)

    Clarke, Mark S. F. (Inventor); Feeback, Daniel L. (Inventor)

    1999-01-01

    A method of staining preselected, mounted specimens of either biological or nonbiological material enclosed within a staining chamber where the liquid staining reagents are applied and removed from the staining chamber using hypergravity as the propelling force. In the preferred embodiment, a spacecraft-operated centrifuge and method of diagnosing biological specimens while in orbit, characterized by hermetically sealing a shell assembly. The assembly contains slide stain apparatus with computer control therefor, the operative effect of which is to overcome microgravity, for example on board an International Space Station.

  12. Adapting lean to histology laboratories.

    Science.gov (United States)

    Buesa, René J

    2009-10-01

    Histology laboratories (histolabs) can increase productivity and reduce turnaround time and errors by using any one of several available management tools. After a few years of operation, all histolabs develop workflow problems. Histology laboratories handling more than 20,000 cases per year benefit the most from implementing management tools, as occurred in the 25 facilities summarized in this article. Discontinuous workflow, lack of "pulling" between steps, accepting unavoidable waiting times while working with small batches within work cells, and a workflow with an uneven rate of completion, are some of the adaptations required by the Lean system when it is used in histology because 70% of the tasks are manual and the flow has to be interrupted to add value to the pieces of tissue during tissue processing, no matter how short that step is. After all these adaptations are incorporated, the histolab becomes as "Lean" as it can be, and the qualifier is also a recognition of the effort and personnel involvement in the implementation. Given its service nature, productivity increments do not expand the histolab customer base and could lead to staffing reductions. This is one of the causes of reluctance by some employees for implementing these techniques which are mostly driven by cost reductions sought by insurance companies and administrators, and not necessarily because of a real medical need to reduce the turnaround time. Finally, any histolab wanting to improve its workflow can follow some easy steps presented here as a guide to accomplish that objective. These steps stress the need for the supervisors to insure that the personnel in the histology laboratory are being paid at a comparable rate as other histolabs in the area.

  13. A novel, modernized Golgi-Cox stain optimized for CLARITY cleared tissue.

    Science.gov (United States)

    Kassem, Mustafa S; Fok, Sandra Y Y; Smith, Kristie L; Kuligowski, Michael; Balleine, Bernard W

    2018-01-15

    High resolution neuronal information is extraordinarily useful in understanding the brain's functionality. The development of the Golgi-Cox stain allowed observation of the neuron in its entirety with unrivalled detail. Tissue clearing techniques, e.g., CLARITY and CUBIC, provide the potential to observe entire neuronal circuits intact within tissue and without previous restrictions with regard to section thickness. Here we describe an improved Golgi-Cox stain method, optimised for use with CLARITY and CUBIC that can be used in both fresh and fixed tissue. Using this method, we were able to observe neurons in their entirety within a fraction of the time traditionally taken to clear tissue (48h). We were also able to show for the first-time that Golgi stained tissue is fluorescent when visualized using a multi-photon microscope, allowing us to image synaptic spines with a detail previously unachievable. These novel methods provide cheap and easy to use techniques to investigate the morphology of cellular processes in the brain at a new-found depth, speed, utility and detail, without previous restrictions of time, tissue type and section thickness. This is the first application of a Golgi-Cox stain to cleared brain tissue, it is investigated and discussed in detail, describing different methodologies that may be used, a comparison between the different clearing techniques and lastly the novel interaction of these techniques with this ultra-rapid stain. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Histological transformations of the dental pulp as possible indicator of post mortem interval: a pilot study.

    Science.gov (United States)

    Carrasco, Patricio A; Brizuela, Claudia I; Rodriguez, Ismael A; Muñoz, Samuel; Godoy, Marianela E; Inostroza, Carolina

    2017-10-01

    The correct estimation of the post mortem interval (PMI) can be crucial on the success of a forensic investigation. Diverse methods have been used to estimate PMI, considering physical changes that occur after death, such as mortis algor, livor mortis, among others. Degradation after death of dental pulp is a complex process that has not yet been studied thoroughly. It has been described that pulp RNA degradation could be an indicator of PMI, however this study is limited to 6 days. The tooth is the hardest organ of the human body, and within is confined dental pulp. The pulp morphology is defined as a lax conjunctive tissue with great sensory innervation, abundant microcirculation and great presence of groups of cell types. The aim of this study is to describe the potential use of pulp post mortem alterations to estimate PMI, using a new methodology that will allow obtainment of pulp tissue to be used for histomorphological analysis. The current study will identify potential histological indicators in dental pulp tissue to estimate PMI in time intervals of 24h, 1 month, 3 months and 6 months. This study used 26 teeth from individuals with known PMI of 24h, 1 month, 3 months or 6 months. All samples were manipulated with the new methodology (Carrasco, P. and Inostroza C. inventors; Universidad de los Andes, assignee. Forensic identification, post mortem interval estimation and cause of death determination by recovery of dental tissue. United State patent US 61/826,558 23.05.2013) to extract pulp tissue without the destruction of the tooth. The dental pulp tissues obtained were fixed in formalin for the subsequent generation of histological sections, stained with Hematoxylin Eosin and Masson's Trichrome. All sections were observed under an optical microscope using magnifications of 10× and 40×. The microscopic analysis of the samples showed a progressive transformation of the cellular components and fibers of dental pulp along PMI. These results allowed creating a

  15. Biochemical and histological evidences for the antitumor potential of ...

    African Journals Online (AJOL)

    Biochemical and histological evidences for the antitumor potential of Teucrium Oliverianum and Rhazya stricta in chemically-induced hepatocellular carcinoma. ... Photomicrograph of liver tissue sections of rats in HCC revealed hepatic parenchyma with foci of anaplastic hepatocellular carcinoma as well as other foci of ...

  16. Remote Histology Learning from Static versus Dynamic Microscopic Images

    Science.gov (United States)

    Mione, Sylvia; Valcke, Martin; Cornelissen, Maria

    2016-01-01

    Histology is the study of microscopic structures in normal tissue sections. Curriculum redesign in medicine has led to a decrease in the use of optical microscopes during practical classes. Other imaging solutions have been implemented to facilitate remote learning. With advancements in imaging technologies, learning material can now be digitized.…

  17. Histological Spectrum of Idiopathic Noncirrhotic Portal Hypertension in Liver Biopsies From Dialysis Patients.

    Science.gov (United States)

    Lee, Hwajeong; Ainechi, Sanaz; Singh, Mandeep; Ells, Peter F; Sheehan, Christine E; Lin, Jingmei

    2015-09-01

    Liver biopsy is performed for various indications in dialysis patients. Being a less-common subset, the hepatic pathology in renal dialysis is not well documented. Idiopathic noncirrhotic portal hypertension (INCPH) is a clinical entity associated with unexplained portal hypertension and/or a spectrum of histopathological vascular changes in the liver. After encountering INCPH and vascular changes of INCPH in 2 renal dialysis patients, we sought to further investigate this noteworthy association. A random search for patients on hemodialysis or peritoneal dialysis with liver biopsy was performed. Hematoxylin and eosin, reticulin, trichrome, and CK7 stains were performed on formalin-fixed, paraffin-embedded tissue sections. Histopathological features were reviewed, and the results were correlated with clinical findings. In all, 13 liver biopsies were retrieved. The mean cumulative duration of dialysis was 50 months (range = 17 months to 11 years). All patients had multiple comorbidities. Indications for biopsy were a combination of abnormal liver function tests (6), portal hypertension (4), ascites (3), and possible cirrhosis (3). Two patients with portal hypertension underwent multiple liver biopsies for diagnostic purposes. All (100%) biopsies showed some histological features of INCPH, including narrowed portal venous lumen (9), increased portal vascular channels (8), shunt vessels (3), dilated sinusoids (9), regenerative nodule (5), and features of venous outflow obstruction (3). No cirrhosis was identified. Liver biopsies from patients on dialysis demonstrate histopathological vascular changes of INCPH. Some (31%) patients present with portal hypertension without cirrhosis. The histological changes may be reflective of underlying risk factors for INCPH in this group. © The Author(s) 2015.

  18. Anterior and posterior compartment 3D endovaginal ultrasound anatomy based on direct histologic comparison.

    Science.gov (United States)

    Shobeiri, S Abbas; White, Dena; Quiroz, Lieschen H; Nihira, Mikio A

    2012-08-01

    We used direct histologic comparison to validate the use of 3D endovaginal ultrasound (EVUS) as a novel and emerging technology for evaluating the structures found in the anterior and posterior pelvic floor compartments. A young nulliparous female pelvis specimen was dissected and histologic slides were prepared by making 8-Micron-thick sagittal cuts. The slides were stained with Mallory trichrome and arranged to form large sections encompassing each anterior and posterior sagittal plane. Healthy nulliparous women underwent 3D EVUS to obtain 3D cubes of the anterior and posterior compartments. Two investigators independently evaluated the anterior and posterior midsagittal structures. The investigators mutually viewed the images and calculated urethral and anal sphincter measurements. Thirty-one nulliparous women underwent 3D EVUS; 77% of the participants were Caucasian, with mean age 31.8 [standard deviation (SD) 5.8] and mean body mass index (BMI) of 28.5 (SD 7.9). The following mean (SD) measurements were obtained: urethral length 36 mm (± 5); striated urogenital sphincter area 0.6 cm(2) (± 0.16); longitudinal and circular smooth muscle area 1.1 cm(2) (± 0.4); urethral complex width 14 mm (± 2); urethral complex area 1.3 cm(2) (± 0.4); internal anal sphincter length 26 mm (± 4); internal anal sphincter thickness 3.2 mm (± 0.8); and rectovaginal septum length 31 mm (± 5). The agreement for visualization of structures was as follows: vesical trigone 96% (κ = 0.65), trigonal ring 94% (κ = 0.8), trigonal plate 84% (κ = 0.6); longitudinal and circular smooth muscle 100%; compressor urethra 97% (κ = 0.85); striated urogenital sphincter 97% (κ = 0.85); rectovaginal septum 100%; internal anal sphincter 100%; external anal sphincter subdivisions 100%. Three-dimensional EVUS can be used to visualize structures of the anterior and posterior compartments in nullipara.

  19. Automated Quantification of Hematopoietic Cell – Stromal Cell Interactions in Histological Images of Undecalcified Bone

    Science.gov (United States)

    Zehentmeier, Sandra; Cseresnyes, Zoltan; Escribano Navarro, Juan; Niesner, Raluca A.; Hauser, Anja E.

    2015-01-01

    Confocal microscopy is the method of choice for the analysis of localization of multiple cell types within complex tissues such as the bone marrow. However, the analysis and quantification of cellular localization is difficult, as in many cases it relies on manual counting, thus bearing the risk of introducing a rater-dependent bias and reducing interrater reliability. Moreover, it is often difficult to judge whether the co-localization between two cells results from random positioning, especially when cell types differ strongly in the frequency of their occurrence. Here, a method for unbiased quantification of cellular co-localization in the bone marrow is introduced. The protocol describes the sample preparation used to obtain histological sections of whole murine long bones including the bone marrow, as well as the staining protocol and the acquisition of high-resolution images. An analysis workflow spanning from the recognition of hematopoietic and non-hematopoietic cell types in 2-dimensional (2D) bone marrow images to the quantification of the direct contacts between those cells is presented. This also includes a neighborhood analysis, to obtain information about the cellular microenvironment surrounding a certain cell type. In order to evaluate whether co-localization of two cell types is the mere result of random cell positioning or reflects preferential associations between the cells, a simulation tool which is suitable for testing this hypothesis in the case of hematopoietic as well as stromal cells, is used. This approach is not limited to the bone marrow, and can be extended to other tissues to permit reproducible, quantitative analysis of histological data. PMID:25938636

  20. Novel Molecular Markers of Malignancy in Histologically Normal and Benign Breast

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    Aejaz Nasir

    2011-01-01

    Full Text Available To detect the molecular changes of malignancy in histologically normal breast (HNB tissues, we recently developed a novel 117-gene-malignancy-signature. Here we report validation of our leading malignancy-risk-genes, topoisomerase-2-alpha (TOP2A, minichromosome-maintenance-protein-2 (MCM2 and “budding-uninhibited-by-benzimidazoles-1-homolog-beta” (BUB1B at the protein level. Using our 117-gene malignancy-signature, we classified 18 fresh-frozen HNB tissues from 18 adult female breast cancer patients into HNB-tissues with low-grade (HNB-LGMA; =9 and high-grade molecular abnormality (HNB-HGMA; =9. Archival sections of additional HNB tissues from these patients, and invasive ductal carcinoma (IDC tissues from six other patients were immunostained for these biomarkers. TOP2A/MCM2 expression was assessed as staining index (% and BUB1B expression as H-scores (0–300. Increasing TOP2A, MCM2, and BUB1B protein expression from HNB-LGMA to HNB-HGMA tissues to IDCs validated our microarray-based molecular classification of HNB tissues by immunohistochemistry. We also demonstrated an increasing expression of TOP2A protein on an independent test set of HNB/benign/reductionmammoplasties, atypical-ductal-hyperplasia with and without synchronous breast cancer, DCIS and IDC tissues using a custom tissue microarray (TMA. In conclusion, TOP2A, MCM2, and BUB1B proteins are potential molecular biomarkers of malignancy in histologically normal and benign breast tissues. Larger-scale clinical validation studies are needed to further evaluate the clinical utility of these molecular biomarkers.

  1. Fluorescent Staining of Tea Pathogenic Fungi in Tea Leaves Using Fluorescein-labeled Lectin

    Science.gov (United States)

    Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

    Fluorochrome-labeled lectin, fluorescein conjugated wheat germ agglutinin (F-WGA) was applied to stain tea pathogenic fungi in tea leaf tissue. Infected leaves were fixed and decolorized with a mixture of ethanol and acetic acid, and cleared with 10% KOH for whole mount before staining with F-WGA. Hyphae of Pestalotiopsis longiseta, Pseudocercospora ocellata, Botrytis cinerea and Colletotrichum theae-sinensis fluoresced brightly in whole mount and sectioned samples of infected leaf tissue. In browned tissue, hyphae did not fluoresce frequently in whole mount sample. Autofluorescence of leaf tissue was strong in browned tissue of sections, it was removed by 10% KOH treatment before staining. Penetration hyphae of C. theae-sinensis in cell wall of trichome and hyphae in basal part of trichome did not fluoresced frequently. In whole mount samples of tea leaf infected with Exobasidium vexans and E. reticulatum, hymenia appeared on leaf surface fluoresced, but hyphae in leaf tissue did not fluoresce. In sectioned samples, hyphae fluoresced brightly when sections were treated with 10% KOH before staining.

  2. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Science.gov (United States)

    2010-01-01

    ...-blood test. 3 147.3 Section 147.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... Blood Testing Procedures § 147.3 The stained-antigen, rapid, whole-blood test. 3 3 The procedure described is a modification of the method reported by Schaffer, MacDonald, Hall, and Bunyea, Jour. Amer. Vet...

  3. Histological description of Cerdocyon thous (Linnaeus, 1766 respiratory system

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    Marla P. Rocha

    Full Text Available ABSTRACT: The massive agricultural expansion converted the Cerdocyon thous, a South American native predator, in vulnerable specie. Basic data, such as histological description, are important to raise awareness on animal species, helping on preservation strategies. Considering the difficult in obtain samples, as the euthanasia of wild animals for this purpose is not allowed, data on histology are very scarce or inexistent. The objective of this paper was to provide a detailed histological description of the trachea and bronchial tree of the crab-eating fox Cerdocyon thous (Linnaeus, 1766. The specimens (one adult male and one adult female used were provided by the Federal University of Pelotas (Pelotas, RS, Brazil Rehabilitation Center of Wild Fauna (NURFS. Tissue samples were fixed in 10% formalin and included in paraffin. After slicing, samples were stained with HE (hematoxylin and eosin, PAS (periodic acid-Schiff and resorcin fuchsin. Trachea had an average diameter of 7.87mm, and approximately 57% of the mucosa ciliated pseudo-stratified columnar epithelium was composed of goblet cells, mostly in the dorsal region. Bronchia and bronchioles had a mucosal fold with higher number of goblet cells. Using all these techniques there is no great remarkable differences from C. thous trachea and lung, when compared with the previous described structures for carnivores and most mammals, except for the goblet cells “regionalization”. Described results are important to understand the animal physiological and behavioral habits, allowing the development of preservation and protection strategies.

  4. Histological study of rat masseter muscle following experimental occlusal alteration.

    Science.gov (United States)

    Nishide, N; Baba, S; Hori, N; Nishikawa, H

    2001-03-01

    It has been suggested that occlusal interference results in masticatory muscle dysfunction. In our previous study, occlusal interference reduced the rat masseter energy level during masticatory movements. The purpose of this study was to investigate the histological alterations of rat masseter muscles following experimental occlusal alteration with unilateral bite-raising. A total of eight male adult Wistar rats were equally divided into control and experimental groups. The experimental rats wore bite-raising splints on the unilateral upper molar. However, 4 weeks after the operation, the anterior deep masseter muscles were removed and then stained for succinic acid dehydrogenase (SDH), haematoxylin eosin (HE) and myofibrillar ATPase. Most of the muscle fibres in experimental rats remained intact, although partial histological changes were observed, such as extended connective tissue, appearance of inflammatory cells in the muscle fibres and existence of muscle fibres with central nuclei and central cores. Moreover, the fibre area-fibre frequency histograms of experimental muscle indicated a broad pattern than that of controls. These results indicated that occlusal interference caused histological changes in masseter muscles and that this may be related to the fact that the masseter energy level was reduced during masticatory movements in unilateral bite-raised rats.

  5. Electrostatic control of the coffee stain effect

    Science.gov (United States)

    Wray, Alex; Papageorgiou, Demetrios; Sefiane, Khellil; Matar, Omar

    2013-11-01

    The ``coffee stain effect,'' as first explained by Deegan et al. 1997, has received a great deal of attention amongst modellers and experimentalists in recent years, perhaps due in part to its obvious casual familiarity. However, it maintains interest because of its intriguing reliance on an interplay of a trio of effects: contact line pinning, inhomogeneous mass flux, and resulting capillarity-driven flow. What is more, the effect, and especially its suppression or reversal, find applications in fields as diverse as sample recovery, mass spectroscopy and the printing of Organic LEDs. We examine the motion a nanoparticle-laden droplet deposited on a precursor film, incorporating the effects of capillarity, concentration-dependent rheology, together with a heated substrate and resultant mass flux and Marangoni effects. We allow the substrate to act as an electrode and incorporate a second electrode above the droplet. The potential difference together with a disparity in electrical properties between the two regions results in electrical (Maxwell) stresses at the interface. We show via lubrication theory and via direct numerical simulations that the ring effect typically observed may be suppressed or augmented via appropriate use of electric fields. EPSRC DTG

  6. Erbium doped stain etched porous silicon

    International Nuclear Information System (INIS)

    Gonzalez-Diaz, B.; Diaz-Herrera, B.; Guerrero-Lemus, R.; Mendez-Ramos, J.; Rodriguez, V.D.; Hernandez-Rodriguez, C.; Martinez-Duart, J.M.

    2008-01-01

    In this work a simple erbium doping process applied to stain etched porous silicon layers (PSLs) is proposed. This doping process has been developed for application in porous silicon solar cells, where conventional erbium doping processes are not affordable because of the high processing cost and technical difficulties. The PSLs were formed by immersion in a HF/HNO 3 solution to properly adjust the porosity and pore thickness to an optimal doping of the porous structure. After the formation of the porous structure, the PSLs were analyzed by means of nitrogen BET (Brunauer, Emmett and Teller) area measurements and scanning electron microscopy. Subsequently, the PSLs were immersed in a saturated erbium nitrate solution in order to cover the porous surface. Then, the samples were subjected to a thermal process to activate the Er 3+ ions. Different temperatures and annealing times were used in this process. The photoluminescence of the PSLs was evaluated before and after the doping processes and the composition was analyzed by Fourier transform IR spectroscopy

  7. EVALUATION OF INHOMOGENEITIES IN HISTOLOGICAL STRUCTURES (CARTILAGE, RETINA

    Directory of Open Access Journals (Sweden)

    Lutz Muche

    2011-05-01

    Full Text Available This paper investigates histological tissues by means of image analysis and spatial statistics. For the quantification of cell frequencies and accumulations two statistical characteristics, intensity function and cluster density, are suggested. The samples are histological sections of human articular cartilage and human retina considered in view of changes during the ageing process. The articular cartilage is characterized by continuous changes of both functions, the cell intensity as well as the clusterization. In contrast, the retina is a trilaminar structure formed in the early embryonic stage without changes by ageing.

  8. The histological and histometrical effects of Urtica dioica extract on rat's prostate hyperplasia.

    Science.gov (United States)

    Moradi, Hamid Reza; Erfani Majd, Naeem; Esmaeilzadeh, Saleh; Fatemi Tabatabaei, Sayed Reza

    2015-01-01

    Benign prostatic hyperplasia (BPH) is a common disease in human that gradual overgrowth of the prostate gland leads to impinge on the urethra with impairment in urinary function. Numerous plants improve uncontrolled growth of the prostate gland and improve urinary tract symptoms associated with BPH. In this study, 25 healthy adult male Wistar rats were divided randomly in five groups: G1 (Control group) received ordinary feed without any treatment, G2 received 10 mg kg(-1) testosterone subcutaneously, G3 received 50 mg kg(-1) nettle root extract orally, G4 received 50 mg kg(-1) nettle root extract orally and 10 mg kg(-1) testosterone, G5 received 10 mg kg(-1) almond oil (Almond oil was used as testosterone solvent) subcutaneously. After six weeks, volume and weight of each lobe were measured and samples were taken. The 5 to 6 µm thickness sections were made using paraffin embedding method and stained by hematoxylin and eosin and periodic acid-Schiff. The results showed that prostate volume and ratio of prostate to body weight were increased significantly in the testosterone. Histological and histometrical results showed that dorsal and lateral type 1 and 2 lobes were not changed significantly but the ventral and anterior lobes have changed significantly. Over all, the nettle root could prevent from some of prostatic hyperplasia effects, so that percentage of folded alveoli in ventral lobe reduced insignificantly.

  9. The histological and histometrical effects of Urtica dioica extract on rat’s prostate hyperplasia

    Science.gov (United States)

    Moradi, Hamid Reza; Erfani Majd, Naeem; Esmaeilzadeh, Saleh; Fatemi Tabatabaei, Sayed Reza

    2015-01-01

    Benign prostatic hyperplasia (BPH) is a common disease in human that gradual overgrowth of the prostate gland leads to impinge on the urethra with impairment in urinary function. Numerous plants improve uncontrolled growth of the prostate gland and improve urinary tract symptoms associated with BPH. In this study, 25 healthy adult male Wistar rats were divided randomly in five groups: G1 (Control group) received ordinary feed without any treatment, G2 received 10 mg kg-1 testosterone subcutaneously, G3 received 50 mg kg-1 nettle root extract orally, G4 received 50 mg kg-1 nettle root extract orally and 10 mg kg-1 testosterone, G5 received 10 mg kg-1 almond oil (Almond oil was used as testosterone solvent) subcutaneously. After six weeks, volume and weight of each lobe were measured and samples were taken. The 5 to 6 µm thickness sections were made using paraffin embedding method and stained by hematoxylin and eosin and periodic acid-Schiff. The results showed that prostate volume and ratio of prostate to body weight were increased significantly in the testosterone. Histological and histometrical results showed that dorsal and lateral type 1 and 2 lobes were not changed significantly but the ventral and anterior lobes have changed significantly. Over all, the nettle root could prevent from some of prostatic hyperplasia effects, so that percentage of folded alveoli in ventral lobe reduced insignificantly. PMID:25992248

  10. HISTOLOGICAL STUDY ON INTERNAL ORGANS DEVELOPMENT OF MANGROVE SNAPPER, Lutjanus argentimaculatus LARVAE

    Directory of Open Access Journals (Sweden)

    Ketut Mahardika

    2012-12-01

    Full Text Available Mangrove snapper, Lutjanus argentimaculatus is one of highly economic value of marine fish commodity. Culture of this species has been developed a few years ago. Basic data on biological aspect is still limited whereas those kinds of information are needed. Histological study to determine development of internal organs of the larvae of mangrove snapper was conducted. Amount of 15-20 larvae each from 1 day after DAH hatching (1 DAH up to 30 days after hatching (30 DAH were collected, fixed in Bouin’s solution for 4-6 hours and then transferred into 70% alcohol. The samples were then embedded in paraffin wax and the sections were stained with haematoxylineosin (H&E. Results showed that the eye retina of the mangrove snapper larvae well developed on 3 DAH, whereas their digestive tract was on 5 DAH. Under this condition, the larvae started feeding as showed by present of natural food tissues in the intestine. Development of digestive tract and other internal organs reached their complete form like in adult fish after 20 DAH.

  11. The Effects of Ceftriaxone on Histology, Histomorphometry, and Histochemistry of Testis and Sperm Characteristics in Mice

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    Shapour Hasanzadeh

    2016-08-01

    Full Text Available Background and Objectives: Ceftriaxone is a third-generation cephalosporin antibiotic, which has a broad-spectrum activity against bacteria. Recently, its adverse effects on the reproductive system, was revealed. The aim of this study was to investigate the adverse effects of ceftriaxone on testicular tissue in adult male mice. Methods: A total of 40 adult male mice were randomly divided into 5 groups: Control group received normal saline; the first and second experimental groups, respectively, received ceftriaxone at doses of 20 and 50mg/kg bw for 7 days; and the third and fourth groups, respectively, received 20 and 50mg/kg bw of the drug for 45 days. After preparation of tissue sections, routine and specific staining was performed, and histological, histomorphometric, and histochemical studies were carried out. The data were analyzed by one-way ANOVA and Tukey’s test. Significance level was considered as p0.05 in the interstitial tissue thickness in all the experimental groups compared to the control group. In the experimental groups, Sudan Black and alkaline phosphatase reactions were intense, while PAS reaction was weak. Conclusion: Ceftriaxone damages testicular structure, which includes loss of balance of the spermatogenic cell population and decrease in the spermatogenesis and spermiogenesis processes.  

  12. Histological responses of cutaneous vascular lesions following photodynamic therapy with talaporfin sodium: a chicken comb model.

    Science.gov (United States)

    Ohshiro, Takafumi; Nakajima, Tatsuo; Ogata, Hisao; Kishi, Kazuo

    2009-09-01

    Mono-L-aspartyl chlorin e6 (Talaporfin sodium) is a novel photosensitizer, and is currently being used in photodynamic therapy for various malignant tumors in combination with irradiation with a 664 nm laser. An interesting characteristic of Talaporfin sodium is that the skin photosensitivity after injection of this agent disappears faster than any other existing photosensitizers. This study examined the vascular events that occurred postirradiation in the chicken comb as a capillary malformation model after photosensitization with Talaporfin sodium. A single intravenous bolus injections of Talaporfin sodium was administered to the chickens, and a 1 cm diameter area of the comb of each animal was irradiated with a 664 nm visible red laser. The gross changes in the chicken combs were recorded for 7-14 days after photodynamic therapy. For the histological examination, HE, PTAH and Azan stained sections were analyzed. All treated chicken combs had blanched after photodynamic therapy. Microscopy demonstrated an absence of erythrocytes and the vessel lumina were obliterated, leaving the normal overlying epidermis completely intact. Concomitantly with selective destruction of the capillaries in the target area, moderate invasion of inflammatory cells and a slight increase in the stroma were observed. In the chicken comb model, photodynamic therapy with Talaporfin sodium effectively achieved selective destruction of the microvasculature while leaving the epidermis intact. Our results strongly suggest that photodynamic therapy with Talaporfin sodium could be a feasible method to treat dermal hypervascular lesions.

  13. An Imaging and Histological Study on Intrahepatic Microvascular Passage of Contrast Materials in Rat Liver

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    Qian Xia

    2017-01-01

    Full Text Available Background. Lipiodol has been applied for decades in transarterial chemoembolization to treat liver malignancies, but its intrahepatic pathway through arterioportal shunt (APS in the liver has not been histologically revealed. This rodent experiment was conducted to provide evidence for the pathway of Lipiodol delivered through the hepatic artery (HA but found in the portal vein (PV and to elucidate the observed unidirectional APS. Methods. Thirty rats were divided into 5 groups receiving systemic or local arterial infusion of red-stained iodized oil (RIO or its hydrosoluble substitute barium sulfate suspension (BSS, or infusion of BSS via the PV, monitored by real-time digital radiography. Histomorphology of serial frozen and paraffin sections was performed and quantified. Results. After HA infusion, RIO and BSS appeared extensively in PV lumens with peribiliary vascular plexus (PVP identified as the responsible anastomotic channel. After PV infusion, BSS appeared predominantly in the PV and surrounding sinusoids and to a much lesser extent in the PVP and HA (P<0.001. Fluid mechanics well explains the one-way-valve phenomenon of APS. Conclusions. Intravascularly injected rat livers provide histomorphologic evidences: (1 the PVP exists in between the HA and PV, which is responsible to the APS of Lipiodol; and (2 the intrahepatic vascular inflow appears HA-PVP-PV unidirectional without a physical one-way valve, which can be postulated by the fluid mechanics.

  14. Histological evaluation of the pulp in teeth from dogs with naturally occurring periodontal disease.

    Science.gov (United States)

    Nemec, Ana; Pavlica, Zlatko; Stiblar-Martincic, Draga; Petelin, Milan; Erzen, Damjan; Crossley, David

    2007-12-01

    The purpose of this investigation was to evaluate the pulp of dog teeth affected by advanced periodontal disease. Histological examination was done on demineralized teeth extracted during clinical treatment of mature, client owned small and medium-size breed dogs with either good periodontal health or with advanced naturally occurring periodontal disease. Routinely stained sections from 5 clinically normal teeth and 22 teeth with advanced periodontitis from dogs between 5 and 12-years of age were examined using light microscopy. The pulp cavities of most teeth were narrow with low cellularity and some fibrosis of the pulp. Findings specific to periodontally affected teeth included acute and chronic pulpitis, vascular congestion, and pulp necrosis. A glomus body was identified in the pulp of one tooth and areas of poorly mineralized cementum were seen in both normal and diseased teeth. Age related changes in dog teeth appear similar to those reported for man and the rat. In addition to age related changes, the pulp of dog teeth with advanced periodontal disease were frequently inflamed or necrotic. This may reflect the advanced periodontitis affecting these teeth or a mechanical effect related to excessive tooth mobility. Further study is required to determine the etiology and significance of these findings and to investigate pulp status in less severely diseased teeth.

  15. Histological-morphometric structure of the liver of some cyprinids (Cyprinidae Rafinesque, 1810 from the Zaporizhzhia reservoir

    Directory of Open Access Journals (Sweden)

    T. Sharamok

    2017-12-01

    Full Text Available Purpose. To investigate the state of the liver of wild carp (Cyprinus carpio Linnaeus, 1758 and bream (Abramis brama Linnaeus, 1758 from sites of the Zaporizhzhia reservoir with different human pressures based on histological and cytometric parameters Methodology.The study was carried out in two zones of the Zaporizhzhia reservoir with different degrees and types of human impact - the lower part, located in the agrarian zone (conditionally "environmentally clean" site, and in the Samara Bay with high content of heavy metals in water. The objects of the study were age-4 bream and wild carp. The fish livers for histological studies were obtained from freshly caught fish (in the spring-summer period by anatomical dissection. Fragments of organs of 0.3 – 0.5 cm were taken for preservation. For the preparation of histological preparations, the liver tissue was stained with hematoxylin and eosin. The pictures of histological preparations were made with the aid of a digital camera connected to the microscope. Histologic sections were investigated with 40X magnification using a microscope with the Sciencelab T500 5.17 M digital camera. Findings. Human impact on the hydroelectric system causes signs of hepatocyte hypertrophy and a number of histopathologies in cyprinids in the Samara Bay. The cytometric analysis of the liver showed that the area of hepatocytes in the bay was larger in comparison with the the individuals of similar age from the lower part of the reservoir by 20 and 10% for the wild carp and bream, respectively. In this case, a 14% increase in the small diameter of cells was observed the in hepatocytes of carp, and a 10% increase in large diameter and a 19% increase in nucleus area was observed in bream. The results of the histological invesatigation of the morphological structure of the liver of cyprinids inhabiting the Samara Bay have revealed a number of pathological changes. Fatty liver dystrophy was observed in 37% of bream and 12

  16. Efficacy test of a toothpaste in reducing extrinsic dental stain

    Science.gov (United States)

    Agustanti, A.; Ramadhani, S. A.; Adiatman, M.; Rahardjo, A.; Callea, M.; Yavuz, I.; Maharani, D. A.

    2017-08-01

    This clinical trial compared the external dental stain reduction achieved by tested toothpaste versus placebo in adult patients. In this double-blind, parallel, randomised clinical trial, 45 female volunteers with a mean age of 20 years old were included. All study subjects front teeth were topically applicated with Silver Diamine Fluoride (SDF) to create external dental stains. Subjects were randomized into test (n=22) and control (n=23) groups. Toothpastes were used for two days to analyse the effects of removing external stains on the labial surfaces of all anterior teeth. VITA Easyshade Advance 4.0 was used to measure dental extrinsic stains changes. The analysis showed statistically significant efficacy of the tested toothpaste in reducing external dental stain caused by SDF, comparing to the placebo toothpaste, after one and two days of usage. The tested toothpaste was effective in reducing dental stain.

  17. Post-staining electroblotting for efficient and reliable peptide blotting.

    Science.gov (United States)

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

    Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.

  18. Gram staining in the diagnosis of acute septic arthritis.

    Science.gov (United States)

    Faraj, A A; Omonbude, O D; Godwin, P

    2002-10-01

    This study aimed at determining the sensitivity and specificity of Gram staining of synovial fluid as a diagnostic tool in acute septic arthritis. A retrospective study was made of 22 patients who had arthroscopic lavage following a provisional diagnosis of acute septic arthritis of the knee joint. Gram stains and cultures of the knee aspirates were compared with the clinical and laboratory parameters, to evaluate their usefulness in diagnosing acute arthritis. All patients who had septic arthritis had pain, swelling and limitation of movement. CRP was elevated in 90% of patients. The incidence of elevated white blood cell count was higher in the group of patients with a positive Gram stain study (60%) as compared to patients with a negative Gram stain study (33%). Gram staining sensitivity was 45%. Its specificity was however 100%. Gram staining is an unreliable tool in early decision making in patients requiring urgent surgical drainage and washout.

  19. Techniques for controlling variability in gram staining of obligate anaerobes.

    Science.gov (United States)

    Johnson, M J; Thatcher, E; Cox, M E

    1995-01-01

    Identification of anaerobes recovered from clinical samples is complicated by the fact that certain gram-positive anaerobes routinely stain gram negative; Peptostreptococcus asaccharolyticus, Eubacterium plautii, Clostridium ramosum, Clostridium symbiosum, and Clostridium clostridiiforme are among the nonconformists with regard to conventional Gram-staining procedures. Accurate Gram staining of American Type Culture Collection strains of these anaerobic bacteria is possible by implementing fixing and staining techniques within a gloveless anaerobic chamber. Under anaerobic conditions, gram-positive staining occurred in all test organisms with "quick" fixing techniques with both absolute methanol and formalin. The results support the hypothesis that, when anaerobic bacteria are exposed to oxygen, a breakdown of the physical integrity of the cell wall occurs, introducing Gram stain variability in gram-positive anaerobes. PMID:7538512

  20. Modified Cajal's trichrome stain as a diagnostic aid in the study of epithelial pathology

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    Karpagaselvi Sanjai

    2017-01-01

    Full Text Available Background: Diagnosis of initial epithelial pathology maybe difficult in Squamous Cell Carcinoma (SCC, Carcinoma In Situ and other atypical epithelial malignancies, under routine Haematoxylin and Eosin (H and E stain. The detection of minor basement membrane alterations in doubtful cases is both time consuming and confusing. Aims: To evaluate efficacy of Modified Cajal's Trichrome Stain (CTS in relation to Haematoxylin and Eosin for study of epithelial dysplasia, carcinoma in situ, micro invasive SCC, frank SCC, and SCC in lymph nodes. Materials and Methods: Formalin-fixed, paraffin-embedded tissue blocks of mild epithelial dysplasia (n = 2, moderate epithelial dysplasia (n = 2, severe epithelial dysplasia (n = 4, carcinoma in situ (n = 1, micro-invasive SCC (n = 4, verrucous carcinoma (n = 1, and frank OSCC (n = 5 were stained with CTS and H&E. The sections were compared based on set histopathological criteria. Results and Conclusion: In SCC cases stained with CTS, invasion into connective tissue and keratin pearls were strikingly evident. Depth of invasion could be more accurately determined. Tumour cells in lymph node were intensely contrasted and easily discernible. Thus, CTS is a good differential stain, clearly delineating the epithelial elements from the connective tissue elements visually. This helps in tracing the basement membrane very clearly. It is an economic, rapid and easy to use method which cannot replace Haematoxylin and Eosin stain in cancer diagnosis, but can definitely be used adjunctive to it. Prompt diagnosis is crucial to effective treatment, and this stain assists in early and rapid diagnosis of cancer.

  1. Diagnosing periprosthetic infection: false-positive intraoperative Gram stains.

    Science.gov (United States)

    Oethinger, Margret; Warner, Debra K; Schindler, Susan A; Kobayashi, Hideo; Bauer, Thomas W

    2011-04-01

    Intraoperative Gram stains have a reported low sensitivity but high specificity when used to help diagnose periprosthetic infections. In early 2008, we recognized an unexpectedly high frequency of apparent false-positive Gram stains from revision arthroplasties. The purpose of this report is to describe the cause of these false-positive test results. We calculated the sensitivity and specificity of all intraoperative Gram stains submitted from revision arthroplasty cases during a 3-month interval using microbiologic cultures of the same samples as the gold standard. Methods of specimen harvesting, handling, transport, distribution, specimen processing including tissue grinding/macerating, Gram staining, and interpretation were studied. After a test modification, results of specimens were prospectively collected for a second 3-month interval, and the sensitivity and specificity of intraoperative Gram stains were calculated. The retrospective review of 269 Gram stains submitted from revision arthroplasties indicated historic sensitivity and specificity values of 23% and 92%, respectively. Systematic analysis of all steps of the procedure identified Gram-stained but nonviable bacteria in commercial broth reagents used as diluents for maceration of periprosthetic membranes before Gram staining and culture. Polymerase chain reaction and sequencing showed mixed bacterial DNA. Evaluation of 390 specimens after initiating standardized Millipore filtering of diluent fluid revealed a reduced number of positive Gram stains, yielding 9% sensitivity and 99% specificity. Clusters of false-positive Gram stains have been reported in other clinical conditions. They are apparently rare related to diagnosing periprosthetic infections but have severe consequences if used to guide treatment. Even occasional false-positive Gram stains should prompt review of laboratory methods. Our observations implicate dead bacteria in microbiologic reagents as potential sources of false-positive Gram

  2. Near-UV laser treatment of extrinsic dental enamel stains.

    Science.gov (United States)

    Schoenly, J E; Seka, W; Featherstone, J D B; Rechmann, P

    2012-04-01

    The selective ablation of extrinsic dental enamel stains using a 400-nm laser is evaluated at several fluences for completely removing stains with minimal damage to the underlying enamel. A frequency-doubled Ti:sapphire laser (400-nm wavelength, 60-nanosecond pulse duration, 10-Hz repetition rate) was used to treat 10 extracted human teeth with extrinsic enamel staining. Each tooth was irradiated perpendicular to the surface in a back-and-forth motion over a 1-mm length using an ∼300-µm-diam 10th-order super-Gaussian beam with fluences ranging from 0.8 to 6.4 J/cm(2) . Laser triangulation determined stain depth and volume removed by measuring 3D surface images before and after irradiation. Scanning electron microscopy evaluated the surface roughness of enamel following stain removal. Fluorescence spectroscopy measured spectra of unbleached and photobleached stains in the spectral range of 600-800 nm. Extrinsic enamel stains are removed with laser fluences between 0.8 and 6.4 J/cm(2) . Stains removed on sound enamel leave behind a smooth enamel surface. Stain removal in areas with signs of earlier cariogenic acid attacks resulted in isolated and randomly located laser-induced, 50-µm-diam enamel pits. These pits contain 0.5-µm diam, smooth craters indicative of heat transfer from the stain to the enamel and subsequent melting and water droplet ejection. Ablation stalling of enamel stains is typically observed at low fluences (Laser ablation of extrinsic enamel stains at 400 nm is observed to be most efficient above 3 J/cm(2) with minimal damage to the underlying enamel. Unsound underlying enamel is also observed to be selectively removed after irradiation. Copyright © 2012 Wiley Periodicals, Inc.

  3. Histology of the first fish

    Science.gov (United States)

    Smith, M.P.; Sansom, I.J.; Repetski, J.E.

    1996-01-01

    THE first description of Anatolepis Bockelie & Fortey was from early Ordovician sediments of Ny Friesland, Spitsbergen1,2, but the genus is now known from many localities in North America and Greenland, ranging in age from the Late Cambrian period to the Early Ordovician3-6. Although initially interpreted as an agnathan fish2,3 that predated other representatives7, this has been widely disputed because the available histological data were unconvincing6,8-10 and the scales fell outside the known morphological range of other accepted early vertebrates9-11. Further doubt was cast upon the vertebrate affinity of Anatolepis when specimens from East Greenland were interpreted as the cuticular fragments of aglaspid arthropods6, although this interpretation has also been refuted12. Here we report on the morphology and histology of large collections of Anatolepis, and demonstrate the presence of dentine, a tissue unique to vertebrates, confirming that the taxon is both a vertebrate and the oldest known fish.

  4. Immunostaining for p16(INK4a) used as a conjunctive tool improves interobserver agreement of the histologic diagnosis of cervical intraepithelial neoplasia

    DEFF Research Database (Denmark)

    Horn, L.C.; Reichert, A.; Oster, A.

    2008-01-01

    The quality of cervical histopathology is critical to cervical cancer prevention, cancer treatment, and research programs. On the basis of the histology results further patient management is determined. However, the diagnostic interpretation of histologic hematoxylin-eosin (H&E)-stained slides is...... immunohistochemistry as an adjunct to conventional H&E-stained specimens thus contributes to a more reproducible diagnosis of cervical intraepithelial neoplasia, and may be a valuable aid for the interpretation of cervical histology Udgivelsesdato: 2008/4......The quality of cervical histopathology is critical to cervical cancer prevention, cancer treatment, and research programs. On the basis of the histology results further patient management is determined. However, the diagnostic interpretation of histologic hematoxylin-eosin (H&E)-stained slides......) immunohistochemistry may increase the performance of pathologists in diagnosing squamous lesions in cervical punch and cone biopsies. When using a consecutive p 16(INK4a)-stained slide in conjunction to the H&E-stained slide, interobserver agreement between 6 pathologists improved significantly for both cervical punch...

  5. Utility of Gram staining for diagnosis of Malassezia folliculitis.

    Science.gov (United States)

    Tu, Wei-Ting; Chin, Szu-Ying; Chou, Chia-Lun; Hsu, Che-Yuan; Chen, Yu-Tsung; Liu, Donald; Lee, Woan-Ruoh; Shih, Yi-Hsien

    2018-02-01

    Malassezia folliculitis (MalF) mimics acne vulgaris and bacterial folliculitis in clinical presentations. The role of Gram staining in rapid diagnosis of MalF has not been well studied. In our study, 32 patients were included to investigate the utility of Gram staining for MalF diagnosis. The final diagnoses of MalF were determined according to clinical presentation, pathological result and treatment response to antifungal agents. Our results show that the sensitivity and specificity of Gram staining are 84.6% and 100%, respectively. In conclusion, Gram staining is a rapid, non-invasive, sensitive and specific method for MalF diagnosis. © 2017 Japanese Dermatological Association.

  6. Efficacy of in-house fluorescent stain for fungus

    Directory of Open Access Journals (Sweden)

    K. R. L. Surya Kirani

    2017-01-01

    Full Text Available Context: Mycotic infections are gaining importance in the present day medicine, and definite demonstration of fungus is essential for diagnosis. Small numbers of organisms in the smear can be identified by fluorescence microscopy. Calcofluor white (CFW fluorescent stain is a textile brightener mixed with Evans blue. It is expensive and not easily available. Aims: (1 To assess the efficacy of in-house CFW fluorescent stain for fungus in relation to conventional CFW stain, histopathology, and culture. (2 To determine sensitivity, specificity, negative predictive value (NPV, and positive predictive value (PPV with culture as gold standard. Settings and Design: One hundred cases of suspected dermatophytosis and 15 cases of systemic mycosis were included in the study. Subjects and Methods: The local whitener Ranipal is added with Robin blue, another brightener, and was used to stain teased fungal cultures. Skin, hair, and nails require pretreatment with potassium hydroxide (KOH. Biopsy slides require deparaffinization and pretreatment with KOH before staining. Conventional calcofluor stain, histopathology, and culture were done. Statistical Analysis Used: Statistical analysis was performed using sensitivity, specificity, NPV, and PPV. Results: The results are consistently comparable with conventional stain. The sensitivity was 100%, specificity was 93.3%, NPV was 100%, and PPV was 85.7%. It is also cost effective when compared to commercial stains. Conclusions: In-house stain can be used for screening of fungus in direct samples, biopsies as alternative in resource-constrained laboratories.

  7. Early colonic dysplasia: comparison of differential mucin staining and tritiated thymidine labeling

    International Nuclear Information System (INIS)

    Chabot, J.A.; Colacchio, T.A.

    1985-01-01

    Controversy has arisen regarding the interpretation and significance of histochemical changes in the mucin produced by the globlet cells in colonic mucosa. The shift from sulfomucin to sialomucin, which is readily identified utilizing high iron diamine-alcian blue staining techniques, has been alternately interpreted as a specific, early dysplastic and premalignant change or a nonspecific generalized response to trauma and inflammation, among others. An attempt to clarify this issue was made by comparing mucin changes identified by high iron diamine-alcian blue staining techniques with increases in DNA synthetic activity identified utilizing autoradiographic analysis of tritiated thymidine uptake. Male Holtzman rats were treated with 15 weekly subcutaneous injections of dimethylhydrazine (30 mg/kg per week) (10 rats) or placebo (10 rats). The colons were prepared and fixed, sequential sections were stained with hematoxylin-eosin or high iron diamine-alcian blue, autoradiography was performed. Analyses of labeling index showed no difference in normal background crypts between the control and treatment groups nor in crypts adjacent to those displaying abnormal mucin staining. Crypts with abnormal mucin production (sialomucin dominant) had significantly higher labeling indexes when compared with those of control animals (p less than 0.005). These findings indicate that the shifts in mucin production identified with high iron diamine-alcian blue staining represent crypts with increased and abnormally distributed mitotic activity that is an early dysplastic response to the carcinogenic stimulus

  8. Osteoarthritis in the Knee Joints of Göttingen Minipigs after Resection of the Anterior Cruciate Ligament? Missing Correlation of MRI, Gene and Protein Expression with Histological Scoring.

    Directory of Open Access Journals (Sweden)

    Gregor Reisig

    Full Text Available The Göttingen Minipig (GM is used as large animal model in articular cartilage research. The aim of the study was to introduce osteoarthritis (OA in the GM by resecting the anterior cruciate ligament (ACLR according to Pond and Nuki, verified by histological and magnetic resonance imaging (MRI scoring as well as analysis of gene and protein expression.The eight included skeletally mature female GM were assessed after ACLR in the left and a sham operation in the right knee, which served as control. 26 weeks after surgery the knee joints were scanned using a 3-Tesla high-field MR tomography unit with a 3 T CP Large Flex Coil. Standard proton-density weighted fat saturated sequences in coronal and sagittal direction with a slice thickness of 3 mm were used. The MRI scans were assessed by two radiologists according to a modified WORMS-score, the X-rays of the knee joints by two evaluators. Osteochondral plugs with a diameter of 4mm were taken for histological examination from either the main loading zone or the macroscopic most degenerated parts of the tibia plateau or condyle respectively. The histological sections were blinded and scored by three experts according to Little et al. Gene expression analysis was performed from surrounding cartilage. Expression of adamts4, adamts5, acan, col1A1, col2, il-1ß, mmp1, mmp3, mmp13, vegf was determined by qRT-PCR. Immunohistochemical staining (IH of Col I and II was performed. IH was scored using a 4 point grading (0-no staining; 3-intense staining.Similar signs of OA were evident both in ACLR and sham operated knee joints with the histological scoring result of the ACLR joints with 6.48 ± 5.67 points and the sham joints with 6.86 ± 5.84 points (p = 0.7953 The MRI scoring yielded 0.34 ± 0.89 points for the ACLR and 0.03 ± 0.17 for the sham knee joints. There was no correlation between the histological and MRI scores (r = 0.10021. The gene expression profiles as well as the immunohistochemical findings

  9. Biofilm detection in chronic rhinosinusitis by combined application of hematoxylin-eosin and gram staining.

    Science.gov (United States)

    Tóth, László; Csomor, Péter; Sziklai, István; Karosi, Tamás

    2011-10-01

    The pathomechanism of chronic rhinosinusitis with nasal polyposis (CRS/NP) seems to be unclear. Bacterial-, fungal- and combined biofilms might play a potential role in the pathogenesis of various inflammatory diseases and recently in CRS/NP. A prospective, blinded observational study was performed to confirm that the combination of conventional hematoxylin-eosin (HE) and Gram staining protocols could be used to detect bacterial and fungal biofilms in patients with CRS/NP. A total of 50 patients with CRS/NP undergoing endoscopic sinus surgery (ESS) were analyzed. The negative control group consisted of 12 patients undergoing septoplasty for nasal obstruction without CRS/NP. The nasal polyps and inferior turbinate mucosa specimens applied as negative controls were processed to HE and Gram staining. Biofilm was detected in 44 of 50 patients with CRS/NP and in none of 12 negative controls. In our series, HE method showed an obvious correlation with the results of Gram staining and was allocated to be a good predictor of biofilm existence. It was found that the microscopic structure and thickness of biofilms were strongly associated with the integrity of nasal mucosa and with the characteristics of subepithelial cellular infiltration. This study confirmed the presence of bacterial and fungal biofilms on the surface of NPs obtained from patients with CRS. Since biofilms may affect the severity and recurrence rate of CRS treated by ESS they should be detected histologically. In conclusion, HE staining combined with Gram protocol is a robust and reliable method for the detection of bacterial and fungal biofilms in CRS/NP.

  10. Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms--which stain is suitable?

    Science.gov (United States)

    Netuschil, Lutz; Auschill, Thorsten M; Sculean, Anton; Arweiler, Nicole B

    2014-01-11

    There is confusion over the definition of the term "viability state(s)" of microorganisms. "Viability staining" or "vital staining techniques" are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data. Many terms describe "vitality states" of microorganisms, however, several of them are misleading. Authors define "viable" as "capable to grow". Accordingly, staining methods are substitutes, since no staining can prove viability.The reliability of a commercial "viability" staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the "viability" kit are dependent on the stains' concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique.To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research.Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported. - The nomenclature regarding "viability" and "vitality" should be used carefully.- The manual of the commercial "viability" kit itself points out that the kit is not suitable for natural multispecies biofilm research, as supported by an

  11. Histologic effects of resurfacing lasers.

    Science.gov (United States)

    Freedman, Joshua R; Greene, Ryan M; Green, Jeremy B

    2014-02-01

    By utilizing resurfacing lasers, physicians can significantly improve the appearance of sun-damaged skin, scars, and more. The carbon dioxide and erbium:yttrium-aluminum-garnet lasers were the first ablative resurfacing lasers to offer impressive results although these earlier treatments were associated with significant downtime. Later, nonablative resurfacing lasers such as the neodymium:yttrium-aluminum-garnet laser proved effective, after a series of treatments with less downtime, but with more modest results. The theory of fractional photothermolysis has revolutionized resurfacing laser technology by increasing the safety profile of the devices while delivering clinical efficacy. A review of the histologic and molecular consequences of the resurfacing laser-tissue interaction allows for a better understanding of the devices and their clinical effects. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  12. Quantitative histological models suggest endothermy in plesiosaurs

    Directory of Open Access Journals (Sweden)

    Corinna V. Fleischle

    2018-06-01

    Full Text Available Background Plesiosaurs are marine reptiles that arose in the Late Triassic and survived to the Late Cretaceous. They have a unique and uniform bauplan and are known for their very long neck and hydrofoil-like flippers. Plesiosaurs are among the most successful vertebrate clades in Earth’s history. Based on bone mass decrease and cosmopolitan distribution, both of which affect lifestyle, indications of parental care, and oxygen isotope analyses, evidence for endothermy in plesiosaurs has accumulated. Recent bone histological investigations also provide evidence of fast growth and elevated metabolic rates. However, quantitative estimations of metabolic rates and bone growth rates in plesiosaurs have not been attempted before. Methods Phylogenetic eigenvector maps is a method for estimating trait values from a predictor variable while taking into account phylogenetic relationships. As predictor variable, this study employs vascular density, measured in bone histological sections of fossil eosauropterygians and extant comparative taxa. We quantified vascular density as primary osteon density, thus, the proportion of vascular area (including lamellar infillings of primary osteons to total bone area. Our response variables are bone growth rate (expressed as local bone apposition rate and resting metabolic rate (RMR. Results Our models reveal bone growth rates and RMRs for plesiosaurs that are in the range of birds, suggesting that plesiosaurs were endotherm. Even for basal eosauropterygians we estimate values in the range of mammals or higher. Discussion Our models are influenced by the availability of comparative data, which are lacking for large marine amniotes, potentially skewing our results. However, our statistically robust inference of fast growth and fast metabolism is in accordance with other evidence for plesiosaurian endothermy. Endothermy may explain the success of plesiosaurs consisting in their survival of the end-Triassic extinction

  13. Automatic registration of multi-modal microscopy images for integrative analysis of prostate tissue sections

    International Nuclear Information System (INIS)

    Lippolis, Giuseppe; Edsjö, Anders; Helczynski, Leszek; Bjartell, Anders; Overgaard, Niels Chr

    2013-01-01

    Prostate cancer is one of the leading causes of cancer related deaths. For diagnosis, predicting the outcome of the disease, and for assessing potential new biomarkers, pathologists and researchers routinely analyze histological samples. Morphological and molecular information may be integrated by aligning microscopic histological images in a multiplex fashion. This process is usually time-consuming and results in intra- and inter-user variability. The aim of this study is to investigate the feasibility of using modern image analysis methods for automated alignment of microscopic images from differently stained adjacent paraffin sections from prostatic tissue specimens. Tissue samples, obtained from biopsy or radical prostatectomy, were sectioned and stained with either hematoxylin & eosin (H&E), immunohistochemistry for p63 and AMACR or Time Resolved Fluorescence (TRF) for androgen receptor (AR). Image pairs were aligned allowing for translation, rotation and scaling. The registration was performed automatically by first detecting landmarks in both images, using the scale invariant image transform (SIFT), followed by the well-known RANSAC protocol for finding point correspondences and finally aligned by Procrustes fit. The Registration results were evaluated using both visual and quantitative criteria as defined in the text. Three experiments were carried out. First, images of consecutive tissue sections stained with H&E and p63/AMACR were successfully aligned in 85 of 88 cases (96.6%). The failures occurred in 3 out of 13 cores with highly aggressive cancer (Gleason score ≥ 8). Second, TRF and H&E image pairs were aligned correctly in 103 out of 106 cases (97%). The third experiment considered the alignment of image pairs with the same staining (H&E) coming from a stack of 4 sections. The success rate for alignment dropped from 93.8% in adjacent sections to 22% for sections furthest away. The proposed method is both reliable and fast and therefore well suited

  14. Hemangiomas versus arterio-venous malformations: Role of elastic stains and mast cell density

    Directory of Open Access Journals (Sweden)

    Pravin Pawane

    2014-01-01

    Full Text Available Context: Vascular anomalies present diagnostic challenges to histopathologists. Mulliken and Glowacki′s classification categorizes vascular lesions into hemangiomas and vascular malformations. Aim: This study explored diagnostic clues in the histomorphology of hemangiomas and vascular malformations. Materials and Methods: A total of 120 cases of benign vascular lesions were retrieved from 12 years period. A total of 94 cases, where complete clinical details and representative paraffin sections were available, were included in this study. Hematoxylin and eosin (H and E stain and Verhoeff′s stain for elastic tissue were done on all cases and lesions were classified into hemangiomas or arterio-venous malformations (AVM. Mast cell density in all lesions was calculated from toluidine blue stained sections. Results: Ten cases of hemangiomas were reclassified as AVM on the basis of presence and absence of arteriovenous structures. Intra-lesional nerves were seen in significantly higher number of AVMs compared to hemangiomas. Medium and thick sized nerve bundles were seen in 56% of AVMs, while they were not seen in any of the hemangiomas. Mean mast cell density was significantly higher in proliferating hemangiomas (53.12 ± 27.83 cells/mm 2 compared to involuting hemangiomas (11.43 ± 7.9 cells/mm 2 . Conclusions: Use of elastic tissue stains are useful ancillary tools to distinguish between AVMs and hemangiomas. The presence of arteries and arterioles are an integral part of AVMs. The presence of the intra-lesional nerves can be useful to distinguish between AVMs and hemangiomas even on H and E stained sections. The significantly higher mast cell density seen in proliferating hemangiomas compared with involuting ones, seem to suggest that mast cells play an important role in the natural history of these lesions.

  15. Alcian blue-stained particles in a eutrophic lake

    DEFF Research Database (Denmark)

    Worm, J.; Søndergaard, Morten

    1998-01-01

    We used a neutral solution of Alcian Blue to stain transparent particles in eutrophic Lake Frederiksborg Slotss0, Denmark. Alcian Blue-stained particles (ABSP) appeared to be similar to the so-called transparent exopolymer particles (TEP) identified with an acidic solution of Alcian Blue. Our...

  16. News from the Biological Stain Commission No. 11

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2012-01-01

    The 11th issue of News from the Biological Stain Commission (BSC) provides our first impressions of the REACH and ECHA programs. We intend to give a more thorough account of what these important programs actually mean in later editions of News from the Biological Stain Commission. Under the heading...

  17. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced a nu...

  18. Lasers or light sources for treating port-wine stains

    DEFF Research Database (Denmark)

    Faurschou, Annesofie; Olesen, Anne Braae; Leonardi-Bee, Jo

    2011-01-01

    Port-wine stains are birthmarks caused by malformations of blood vessels in the skin. Port-wine stains manifest themselves in infancy as a flat, red mark and do not regress spontaneously but may, if untreated, become darker and thicker in adult life. The profusion of various lasers and light...

  19. Cellient™ automated cell block versus traditional cell block preparation: a comparison of morphologic features and immunohistochemical staining.

    Science.gov (United States)

    Wagner, David G; Russell, Donna K; Benson, Jenna M; Schneider, Ashley E; Hoda, Rana S; Bonfiglio, Thomas A

    2011-10-01

    Traditional cell block (TCB) sections serve as an important diagnostic adjunct to cytologic smears but are also used today as a reliable preparation for immunohistochemical (IHC) studies. There are many ways to prepare a cell block and the methods continue to be revised. In this study, we compare the TCB with the Cellient™ automated cell block system. Thirty-five cell blocks were obtained from 16 benign and 19 malignant nongynecologic cytology specimens at a large university teaching hospital and prepared according to TCB and Cellient protocols. Cell block sections from both methods were compared for possible differences in various morphologic features and immunohistochemical staining patterns. In the 16 benign cases, no significant morphologic differences were found between the TCB and Cellient cell block sections. For the 19 malignant cases, some noticeable differences in the nuclear chromatin and cellularity were identified, although statistical significance was not attained. Immunohistochemical or special stains were performed on 89% of the malignant cases (17/19). Inadequate cellularity precluded full evaluation in 23% of Cellient cell block IHC preparations (4/17). Of the malignant cases with adequate cellularity (13/17), the immunohistochemical staining patterns from the different methods were identical in 53% of cases. The traditional and Cellient cell block sections showed similar morphologic and immunohistochemical staining patterns. The only significant difference between the two methods concerned the lower overall cell block cellularity identified during immunohistochemical staining in the Cellient cell block sections. Copyright © 2010 Wiley-Liss, Inc.

  20. Decreased mortality associated with prompt Gram staining of blood cultures.

    Science.gov (United States)

    Barenfanger, Joan; Graham, Donald R; Kolluri, Lavanya; Sangwan, Gaurav; Lawhorn, Jerry; Drake, Cheryl A; Verhulst, Steven J; Peterson, Ryan; Moja, Lauren B; Ertmoed, Matthew M; Moja, Ashley B; Shevlin, Douglas W; Vautrain, Robert; Callahan, Charles D

    2008-12-01

    Gram stains of positive blood cultures are the most important factor influencing appropriate therapy. The sooner appropriate therapy is initiated, the better. Therefore, it is reasonable to expect that the sooner Gram stains are performed, the better. To determine the value of timely Gram stains and whether improvement in Gram stain turnaround time (TAT) is feasible, we compared data for matched pairs of patients with cultures processed promptly ( or =1 hour TAT) and then monitored TAT by control charting.In 99 matched pairs, average difference in time to detection of positive blood cultures within a pair of patients was less than 0.1 hour. For the less than 1 hour TAT group, the average TAT and crude mortality were 0.1 hour and 10.1%, respectively; for the 1 hour or longer TAT group, they were 3.3 hours and 19.2%, respectively (P Gram stains.

  1. A vocabulary for the identification and delineation of teratoma tissue components in hematoxylin and eosin-stained samples

    Directory of Open Access Journals (Sweden)

    Ramamurthy Bhagavatula

    2014-01-01

    Full Text Available We propose a methodology for the design of features mimicking the visual cues used by pathologists when identifying tissues in hematoxylin and eosin (H&E-stained samples. Background: H&E staining is the gold standard in clinical histology; it is cheap and universally used, producing a vast number of histopathological samples. While pathologists accurately and consistently identify tissues and their pathologies, it is a time-consuming and expensive task, establishing the need for automated algorithms for improved throughput and robustness. Methods: We use an iterative feedback process to design a histopathology vocabulary (HV, a concise set of features that mimic the visual cues used by pathologists, e.g. "cytoplasm color" or "nucleus density." These features are based in histology and understood by both pathologists and engineers. We compare our HV to several generic texture-feature sets in a pixel-level classification algorithm. Results: Results on delineating and identifying tissues in teratoma tumor samples validate our expert knowledge-based approach. Conclusions: The HV can be an effective tool for identifying and delineating teratoma components from images of H&E-stained tissue samples.

  2. Significance of bone marrow histology in the diagnosis of acute myeloid leukemia

    International Nuclear Information System (INIS)

    Younis, U.; Saba, K.; Aijaz, J.; Bukhari, M.H.; Naeem, S.

    2011-01-01

    Acute Myeloid Leukemia (AML) is a heterogeneous disease. The precise diagnosis requires a careful morphological examination of a well pre-pared bone marrow aspirate along with flow cytometry and genetic analysis wherever required. Traditionally, bone marrow biopsy has not been considered an essential diagnostic modality for AML. The aim of this study was to assess the diagnostic as well as prognostic significance of bone marrow histology in patient with acute myeloid leukemia. Forty (40) patients of AML underwent a bone marrow examination including an aspirate and a trephine biopsy. Air dried films of peripheral blood and aspirates were fixed in methanol and stained with Giemsa. The following cytochemical stains were also applied: PAS, Myeloperoxidase, Non specific esterase, Chloracetate Esterase and Acid Phosphatase, and SBB. Bone marrow biopsy specimens were obtained from post superior iliac crest with a manual trephine and were processed in plastic after decalcification. Results: In all the cases there were better diagnostic clues through histological examination of bone marrow particularly in assessing the cellularity, degree of fibrosis, extent of blast infiltration, percentage of inflammatory cells, dysplastic changes and residual haematopoiesis. All these features were better noted in histological examination of core biopsy. The histological examination provided information additional to that provided by aspirate smears about the bone marrow changes in AML and suggested that some of the features may also have pro-gnostic significance in addition to diagnostic importance. (author)

  3. A histological atlas of the tissues and organs of neotenic and metamorphosed axolotl.

    Science.gov (United States)

    Demircan, Turan; İlhan, Ayşe Elif; Aytürk, Nilüfer; Yıldırım, Berna; Öztürk, Gürkan; Keskin, İlknur

    2016-09-01

    Axolotl (Ambystoma Mexicanum) has been emerging as a promising model in stem cell and regeneration researches due to its exceptional regenerative capacity. Although it represents lifelong lasting neoteny, induction to metamorphosis with thyroid hormones (THs) treatment advances the utilization of Axolotl in various studies. It has been reported that amphibians undergo anatomical and histological remodeling during metamorphosis and this transformation is crucial for adaptation to terrestrial conditions. However, there is no comprehensive histological investigation regarding the morphological alterations of Axolotl organs and tissues throughout the metamorphosis. Here, we reveal the histological differences or resemblances between the neotenic and metamorphic axolotl tissues. In order to examine structural features and cellular organization of Axolotl organs, we performed Hematoxylin & Eosin, Luxol-Fast blue, Masson's trichrome, Alcian blue, Orcein and Weigart's staining. Stained samples from brain, gallbladder, heart, intestine, liver, lung, muscle, skin, spleen, stomach, tail, tongue and vessel were analyzed under the light microscope. Our findings contribute to the validation of the link between newly acquired functions and structural changes of tissues and organs as observed in tail, skin, gallbladder and spleen. We believe that this descriptive work provides new insights for a better histological understanding of both neotenic and metamorphic Axolotl tissues. Copyright © 2016 Elsevier GmbH. All rights reserved.

  4. Early human bone response to laser metal sintering surface topography: a histologic report.

    Science.gov (United States)

    Mangano, Carlo; Piattelli, Adriano; d'Avila, Susana; Iezzi, Giovanna; Mangano, Francesco; Onuma, Tatiana; Shibli, Jamil Awad

    2010-01-01

    This histologic report evaluated the early human bone response to a direct laser metal sintering implant surface retrieved after a short period of healing. A selective laser sintering procedure using a Ti-6Al-4V alloy powder with a particle size of 25-45 microm prepared this surface topography. One experimental microimplant was inserted into the anterior mandible of a patient during conventional implant surgery of the jaw. The microimplant and surrounding tissues were removed after 2 months of unloaded healing and were prepared for histomorphometric analysis. Histologically, the peri-implant bone appeared in close contact with the implant surface, whereas marrow spaces could be detected in other areas along with prominently stained cement lines. The mean of bone-to-implant contact was 69.51%. The results of this histologic report suggest that the laser metal sintering surface could be a promising alternative to conventional implant surface topographies.

  5. Classification of breast cancer histology images using Convolutional Neural Networks.

    Directory of Open Access Journals (Sweden)

    Teresa Araújo

    Full Text Available Breast cancer is one of the main causes of cancer death worldwide. The diagnosis of biopsy tissue with hematoxylin and eosin stained images is non-trivial and specialists often disagree on the final diagnosis. Computer-aided Diagnosis systems contribute to reduce the cost and increase the efficiency of this process. Conventional classification approaches rely on feature extraction methods designed for a specific problem based on field-knowledge. To overcome the many difficulties of the feature-based approaches, deep learning methods are becoming important alternatives. A method for the classification of hematoxylin and eosin stained breast biopsy images using Convolutional Neural Networks (CNNs is proposed. Images are classified in four classes, normal tissue, benign lesion, in situ carcinoma and invasive carcinoma, and in two classes, carcinoma and non-carcinoma. The architecture of the network is designed to retrieve information at different scales, including both nuclei and overall tissue organization. This design allows the extension of the proposed system to whole-slide histology images. The features extracted by the CNN are also used for training a Support Vector Machine classifier. Accuracies of 77.8% for four class and 83.3% for carcinoma/non-carcinoma are achieved. The sensitivity of our method for cancer cases is 95.6%.

  6. Curcuma longa extract as a histological dye for collagen fibres and red blood cells

    Science.gov (United States)

    Avwioro, O G; Onwuka, S K; Moody, J O; Agbedahunsi, J M; Oduola, T; Ekpo, O E; Oladele, A A

    2007-01-01

    Crude ethanolic extract and column chromatographic fractions of the Allepey cultivar of Curcuma longa Roxb, commonly called turmeric (tumeric) in commerce, were used as a stain for tissue sections. Staining was carried out under basic, acidic and neutral media conditions. Inorganic and organic dissolution solvents were used. The stain was used as a counterstain after alum and iron haematoxylins. C. longa stained collagen fibres, cytoplasm, red blood cells and muscle cells yellow. It also stained in a fashion similar to eosin, except for its intense yellow colour. Preliminary phytochemical evaluation of the active column fraction revealed that it contained flavonoids, free anthraquinone and deoxy sugar. A cheap, natural dye can thus be obtained from C. longa. PMID:17451535

  7. Mercury localization in mouse kidney over time: autoradiography versus silver staining

    International Nuclear Information System (INIS)

    Rodier, P.M.; Kates, B.; Simons, R.

    1988-01-01

    Several methods of silver staining have been employed to localize mercury in tissue, under the assumption that the techniques represent total Hg, but recent reports have suggested that these stains are specific for a limited fraction of the Hg present in some samples. Magos et al. hypothesized that the stains actually vary with inorganic mercury content. The purpose of the present study was to compare localization by radiolabeling to localization by one silver stain, the photoemulsion histochemical technique, in tissues prepared to contain a range of levels of total Hg and a range of levels of inorganic Hg. Mice dosed with 8 mg Hg/kg as MeHg were killed 24 hr, 1 week, or 2 weeks after exposure, to allow a decrease in total Hg and an increase in the proportion of demethylated Hg over time. Mice dosed with 4 mg Hg/kg as HgCl 2 provided samples in which all the Hg present was in the inorganic form. Atomic absorption of kidneys of mice dosed with MeHg showed that total Hg fell from 55 micrograms/g to 39 to 25 over 2 weeks, while the inorganic fraction climbed from about 2 to 27 to 35%. Grain counts from autoradiographs of 203 Hg-labeled sections correlated with total Hg content at +0.88, but silver staining was correlated with inorganic Hg content, appearing only at late termination times in MeHg-exposed animals, but soon after dosing in mice exposed to inorganic Hg. The photoemulsion histochemical technique revealed a substance strictly localized in the proximal tubules, while autoradiographs and grain counts showed total Hg to be present throughout the kidney tissue. These results support the contention that silver stains are selective for inorganic Hg

  8. PIONEERING WORK IN COMPUTER VISION FOR HISTOLOGY

    OpenAIRE

    Andrei Daniel Timofte; Irina-Draga Căruntu

    2017-01-01

    The concept of computer-assisted training was firstly implemented in UMF Iasi in 1997, through the development of two pioneering digital instruments, namely the Histology Album and the Oral Histology Album. Both were built for learning purposes and became available, at that time, as Internet resources worldwide visible. The application of this particular e-learning method was dictated by the importance of well-defined visual images in acquiring basic histologic principles. Thus, t...

  9. Detection of alkaline phosphatase in canine cells previously stained with Wright-Giemsa and its utility in differentiating osteosarcoma from other mesenchymal tumors.

    Science.gov (United States)

    Ryseff, Julia K; Bohn, Andrea A

    2012-09-01

    Osteosarcoma (OSA) is a common primary bone tumor in dogs. Demonstration of alkaline phosphatase (ALP) reactivity by tumor cells on unstained slides is useful in differentiating osteosarcoma from other types of sarcoma. However, unstained slides are not always available. The objectives of this study were to evaluate the diagnostic utility of detecting ALP expression in differentiating osteosarcoma from other sarcomas in dogs using cytologic material previously stained with Wright-Giemsa stain and to assess the sensitivity and specificity of ALP expression for diagnosing osteosarcoma using a specific protocol. Archived aspirates of histologically confirmed sarcomas in dogs that had been previously stained with Wright-Giemsa stain were treated with 5-bromo, 4-chloro, 3-indolyl phosphate/nitroblue tetrazolium (BCIP/NBT) as a substrate for ALP. Cells were evaluated for expression of ALP after incubation with BCIP/NBT for 1 hour. Sensitivity and specificity of ALP expression for diagnosis of OSA were calculated. In samples from 83 dogs, cells from 15/17 OSAs and from 4/66 tumors other than OSA (amelanotic melanoma, gastrointestinal stromal tumor, collision tumor, and anaplastic sarcoma) expressed ALP. Sensitivity and specificity of ALP expression detected using BCIP/NBT substrate applied to cells previously stained with Wright-Giemsa stain for OSA were 88 and 94%, respectively. ALP expression detected using BCIP/NBT substrate applied to previously stained cells is useful in differentiating canine OSA from other mesenchymal neoplasms. © 2012 American Society for Veterinary Clinical Pathology.

  10. Amnioinfusion for meconium-stained liquor in labour.

    Science.gov (United States)

    Hofmeyr, G Justus; Xu, Hairong; Eke, Ahizechukwu C

    2014-01-23

    Amnioinfusion is thought to dilute meconium present in the amniotic fluid and so reduce the risk of meconium aspiration. To assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (1 December 2013). Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium staining of the amniotic fluid. Three review authors independently assessed eligibility and trial quality, and extracted data. Fourteen studies of variable quality (4435 women) are included.Subgroup analysis was performed for studies from settings with limited facilities to monitor the baby's condition during labour and intervene effectively, and settings with standard peripartum surveillance.Settings with standard peripartum surveillance: there was considerable heterogeneity for several outcomes. There was no significant reduction in the primary outcomes meconium aspiration syndrome, perinatal death or severe morbidity, and maternal death or severe morbidity. There was a reduction in caesarean sections (CSs) for fetal distress but not overall. Meconium below the vocal cords diagnosed by laryngoscopy was reduced, as was neonatal ventilation or neonatal intensive care unit admission, but there was no significant reduction in perinatal deaths or other morbidity. Planned sensitivity analysis excluding trials with greater risk of bias resulted in an absence of benefits for any of the outcomes studied.Settings with limited peripartum surveillance: three studies were included. In the amnioinfusion group there was a reduction in CS for fetal distress and overall; meconium aspiration syndrome (three studies, 1144 women; risk ratio (RR) 0.17, 95% confidence interval (CI) 0.05 to 0.52); perinatal mortality (three studies, 1151 women; RR 0.24, 95% CI 0.11 to 0.53) and neonatal ventilation or neonatal intensive care unit admission. In one of the studies, meconium

  11. Histologic evaluation of pulpal response to MTA and capsaicin in cats

    Directory of Open Access Journals (Sweden)

    Zahrasadat Madani

    2012-09-01

    Full Text Available Introduction: In direct pup capping, the exposed pulp is directly capped with a capping material to provoke a dentinal bridge formation to seal the exposed area.The aim of this study was to evaluate the pulpal response to MTA and capsaicin in cats. Methods: The sample for this experimental study consisted of 24 canines of cats, weighed approximately 3-4kg. After sedation, the teeth were cut and exposed at 3mm above cementoenamel junction, then divided into two groups: 1 direct pulp capping with MTA 2 direct pulp capping with Capsaicin. The cavities were filled with glass ionomer cement (Fuji IX. The cats were sacrificed in the first, second, and fourth weeks 4 canine teeth were extracted for the purpose of histologic analysis. 6Serial sections were cut parallel to the longitudinal axis of the canines. The sections were stained with hematoxylin & eosin, and were observed and analyzed using a light microscope. Results: In both groups treated with MTA and capsaicin, the inflammation decreased during weeks1, 2, and 4. Less inflammation was seen in MTA group but there was no statistical difference between the two groups (p=0.22. In the group treated with capsaicin, necrosis was observed in every 12 samples, but in the other group treated with MTA, only 1 pulp necrosis was seen in the first week (p=0.000. There was no significant difference in dentinal barrier formation, inflammatory response, and soft tissue changes between the two groups. Conclusions: The results of this study showed that capsaicin can decrease the severity of inflammation, but it is ineffective in dentinal barrier formation.

  12. The Effects of Maternal Hyperthyroidism on Histologic Changes in Parietal Lobe in Rat Embryos

    Directory of Open Access Journals (Sweden)

    Fatemeh Mirsafi

    2017-05-01

    Full Text Available Background Maternal hyperthyroidism causes developmental defects on the nervous system of fetuses. Objectives The present study was designed to study the effects of maternal hyperthyroidism on the development of the parietal lobe in the brain of rat embryos. Methods In this experimental study, thirty Sprague-Dawley rats were randomly divided into three groups. The control group received no injections, the sham group received intraperitoneal injections of distilled water solution containing salt and polysorbate (solvent of levothyroxine, and the experimental group received once-daily, intraperitoneal injections of 0.5 mg/kg levothyroxine for a 10-day period to become hyperthyroid rats. The hyperthyroid rats were then mated, and all pregnant rats were killed on the 20th day of gestation. Fetuses were removed, fixed, and processed for histological procedures. The fetuses were sagitally sectioned at 5 µ thickness and stained with hematoxylin-eosin (H and E technique. The sections were examined using a light microscope and Motic software. Results The results showed no significant difference in the studied variables between the sham and control groups. A significantly increase in body weight and a significant decrease in crown-rump length of embryos was observed in the experimental group when compared to the control group. The mean total thickness of the parietal cortex, ventricular layer, and intermediate layer of embryos showed a significant decrease in the experimental group compared to the control and sham groups. The mean number of cells also showed a significant decrease in the intermediate and ventricular layers in the experimental group compared to the control and sham groups. Conclusions This study showed that maternal hyperthyroidism leads to a reduction in development of the parietal cortex in embryos. Maternal hyperthyroidism can disturb the growth and development of embryos.

  13. Histological and Immunohistochemical Revision of Hepatocellular Adenomas: A Learning Experience

    Directory of Open Access Journals (Sweden)

    S. Fonseca

    2013-01-01

    Full Text Available Light has been shed on the genotype/phenotype correlation in hepatocellular adenoma (HCA recognizing HNF1α-inactivated HCA (H-HCA, inflammatory HCA (IHCA, and β-catenin-activated HCA (b-HCA. We reviewed retrospectively our surgical HCA series to learn how to recognize the different subtypes histopathologically and how to interpret adequately their immunohistochemical staining. From January 1992 to January 2012, 37 patients underwent surgical resection for HCA in our institution. Nine had H-HCA (25% characterized by steatosis and loss of L-FABP expression; 20 had IHCA (55.5% showing CRP and/or SAA expression, sinusoidal dilatation, and variable inflammation; and 1 patient had both H-HCA and IHCA. In 5 patients (14%, b-HCA with GS and β-catenin nuclear positivity was diagnosed, two already with hepatocellular carcinoma. Two cases (5.5% remained unclassified. One of the b-HCA showed also the H-HCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/HNF1α-inactivated HCA, another b-HCA exhibited the IHCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/inflammatory HCA. Interestingly, three patients had underlying vascular abnormalities. Using the recently published criteria enabled us to classify histopathologically our retrospective HCA surgical series with accurate recognition of b-HCA for which we confirm the higher risk of malignant transformation. We also underlined the association between HCA and vascular abnormalities.

  14. Anatomy and histology of the transverse humeral ligament.

    Science.gov (United States)

    Snow, Brian J; Narvy, Steven J; Omid, Reza; Atkinson, Roscoe D; Vangsness, C Thomas

    2013-10-01

    The classic literature describes the transverse humeral ligament (THL) as a distinct anatomic structure with a role in biceps tendon stability; however, recent literature suggests that it is not a distinct anatomic structure. The purpose of this study was to evaluate the gross and microscopic anatomy of the THL, including a specific investigation of the histology of this ligament. Thirty frozen, embalmed cadaveric specimens were dissected to determine the gross anatomy of the THL. Seven specimens were evaluated histologically for the presence of mechanoreceptors and free nerve endings. Two tissue layers were identified in the area described as the THL. In the deep layer, fibers of the subscapularis tendon were found to span the bicipital groove with contributions from the coracohumeral ligament and the supraspinatus tendon. Superficial to this layer was a fibrous fascial covering consisting of distinct bands of tissue. Neurohistology staining revealed the presence of free nerve endings but no mechanoreceptors. This study's findings demonstrate that the THL is a distinct structure continuous with the rotator cuff tendons and the coracohumeral ligament. The finding of free nerve endings in the THL suggests a potential role as a shoulder pain generator. Copyright 2013, SLACK Incorporated.

  15. Effect of Melamine Sponge on Tooth Stain Removal.

    Science.gov (United States)

    Otsuka, Takero; Kawata, Toshitsugu

    2015-01-01

    To investigate the stain removal ability of melamine sponge before aesthetic tooth whitening in extracted teeth. Melamine sponge of thickness 40 mm was compressed and the destruction of the partition wall structure during the compression process was examined under a stereoscopic microscope. An extracted human tooth was cleaned by normal polishing or with melamine sponge for 90 s. To evaluate the stain level, the tooth surfaces were photographed under a stereoscopic microscope at 0, 30, 60 and 90 s. The residual stained region was traced in a high-magnification photograph, and the stain intensity was presented as a change, relative to the intensity before the experiment (0 s). Mechanical cleaning by toothbrushing produced polishing scratches on the tooth surface, whereas use of the melamine sponge resulted in only minimal scratches. As the compression level increased, the stain-removing effect tended to become stronger. Melamine sponge can remove stains from the tooth surface more effectively and less invasively compared to a conventional toothbrush. As no new scratches are made on the tooth surface when using a melamine sponge brush, the risk of re-staining is reduced. Cleaning using a melamine sponge brush can be easily and effectively performed at home and in a dental office.

  16. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    Science.gov (United States)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  17. [Usefulness of sputum Gram staining in community-acquired pneumonia].

    Science.gov (United States)

    Sato, Tadashi; Aoshima, Masahiro; Ohmagari, Norio; Tada, Hiroshi; Chohnabayashi, Naohiko

    2002-07-01

    To evaluate the usefulness of sputum gram staining in community-acquired pneumonia (CAP), we reviewed 144 cases requiring hospitalization in the last 4 years. The sensitivity was 75.5%, specificity 68.2%, positive predictive value 74.1%, negative predictive value 69.8%, positive likelihood ratio 2.37, negative likelihood ratio 0.36 and accuracy 72.2% in 97 cases. Both sputum gram staining and culture were performed. Concerning bacterial pneumonia (65 cases), we compared the Gram staining group (n = 33), which received initial antibiotic treatment, based on sputum gram staining with the Empiric group (n = 32) that received antibiotics empirically. The success rates of the initial antibiotic treatment were 87.9% vs. 78.1% (P = 0.473); mean hospitalization periods were 9.67 vs. 11.75 days (P = 0.053); and periods of intravenous therapy were 6.73 vs. 7.91 days (P = 0.044), respectively. As for initial treatment, penicillins were used in the Gram staining group more frequently (P gram staining is useful for the shortening of the treatment period and the appropriate selection of initial antibiotics in bacterial pneumonia. We believe, therefore, that sputum gram staining is indispensable as a diagnostic tool CAP.

  18. Improved method for combination of immunocytochemistry and Nissl staining.

    Science.gov (United States)

    Kádár, Andrea; Wittmann, Gábor; Liposits, Zsolt; Fekete, Csaba

    2009-10-30

    Nissl staining is a widely used method to study morphology and pathology of neural tissue. After standard immunocytochemistry, the Nissl staining labels only the nucleus of neurons and the characteristic staining of the neuronal perikarya is absent or very weak. We hypothesized that the RNA degradation during the immunocytochemical treatment results in the loss of cytoplasmic staining with Nissl-dyes. To test this hypothesis, we used RNAse-free conditions for all steps of immunostaining. To further prevent the RNA-degradation by RNAse contaminations, the RNAse inhibitor heparin was added to all antibody-containing solutions. The efficiency of Nissl staining after standard and RNAse-free double-labeling immunocytochemistry was compared using antibodies against c-Fos and neuropeptide Y (NPY) on tissues of rats refed after 3 days of fasting. After standard immunocytochemistry, the Nissl-staining labeled the nuclei of neurons and only very faintly the cytoplasm of these cells. The RNAse-free treatment did not alter the distribution of immunoreaction signal, but preserved the staining of neuronal perikarya by the Nissl-dyes. In conclusion, the RNAse-free conditions during immunocytochemistry allow the labeling of neuronal perikarya by Nissl-dyes. The described method facilitates the mapping of immunocytochemical signals and makes possible the light microscopic examination of the innervation of neurons identified by their nuclear protein content.

  19. Silver and Cyanine Staining of Oligonucleotides in Polyacrylamide Gel.

    Science.gov (United States)

    Tang, Weizhong; Zhou, Huafu; Li, Wei

    2015-01-01

    To explore why some oligonucleotides in denaturing polyacrylamide gel could not be silver-stained, 134 different oligonucleotides were analyzed using denaturing polyacrylamide gel electrophoresis stained with silver and asymmetric cyanine. As a result, we found that the sensitivity of oligos (dA), (dC), (dG) and (dT) to silver staining could be ranged as (dA) > (dG) > (dC) > (dT) from high to low. It was unexpected that oligo (dT) was hard to be silver-stained. Moreover, the silver staining of an oligonucleotide containing base T could be partially or completely inhibited by base T. The inhibition of silver staining by base T was a competitive inhibition which could be affected by the amounts of the argyrophil nucleobase and base T, the cis-distance between the argyrophil nucleobase and base T, and the gel concentration. The changes of the intensity of an oligonucleotide band caused by the changes of DNA base composition were diverse and interesting. The intensity of some oligonucleotide bands would significantly change when the changes of DNA base composition accumulated to a certain extent (usually ≥ 4 nt). The sensitivity of cyanine staining of ≤ 11-nt long oligonucleotides could be enhanced about 250-fold by fixing the gels with methanol fixing solution.

  20. A novel washing algorithm for underarm stain removal

    Science.gov (United States)

    Acikgoz Tufan, H.; Gocek, I.; Sahin, U. K.; Erdem, I.

    2017-10-01

    After contacting with human sweat which comprise around 27% sebum, anti-perspirants comprising aluminium chloride or its compounds form a jel-like structure whose solubility in water is very poor. In daily use, this jel-like structure closes sweat pores and hinders wetting of skin by sweat. However, when in contact with garments, they form yellowish stains at the underarm of the garments. These stains are very hard to remove with regular machine washing. In this study, first of all, we focused on understanding and simulating such stain formation on the garments. Two alternative procedures are offered to form jel-like structures. On both procedures, commercially available spray or deo-stick type anti-perspirants, standard acidic and basic sweat solutions and artificial sebum are used to form jel-like structures, and they are applied on fabric in order to get hard stains. Secondly, after simulation of the stain on the fabric, we put our efforts on developing a washing algorithm specifically designed for removal of underarm stains. Eight alternative washing algorithms are offered with varying washing temperature, amounts of detergent, and pre-stain removal procedures. Better algorithm is selected by comparison of Tristimulus Y values after washing.

  1. Darkfield illumination improves microscopic detection of metals in Timm's stained tissue

    DEFF Research Database (Denmark)

    Baatrup, E; Frederickson, C J

    1989-01-01

    Deposits of trace or toxic metals can be quickly identified by light microscopical surveys of tissue sections stained for metals by variants of Timm's silver enhancement method. The present work shows that the small, isolated silver grains that label isolated deposits of metal in tissue are undet...... are undetectable in brightfield light microscopy but are easily detected in darkfield microscopy. Darkfield illumination is therefore recommended for improving the detection of trace or toxic metals in tissue. Udgivelsesdato: 1989-Aug......Deposits of trace or toxic metals can be quickly identified by light microscopical surveys of tissue sections stained for metals by variants of Timm's silver enhancement method. The present work shows that the small, isolated silver grains that label isolated deposits of metal in tissue...

  2. Osteoarthritis - histology and pathogenetic concepts

    International Nuclear Information System (INIS)

    Sulzbacher, I.

    2000-01-01

    Osteoarthitis is the most common joint disease affecting over 60% of the elderly population. It is characterized by the progressive erosion of articular cartilage leading to incapacity of movement. In the great majority of instances, osteoarthritis appears insidiously, without apparent initiating cause. This primary form is usually oligoarticular involving hip, knee, cervical vertebrae, interphalangeal joints of the fingers or tarsometatarsal joints of the feet. In case of an underlying systemic diasease or local injury the cartilage destruction is considered as secondary osteoarthritis. The pathogenesis of primary osteoarthritis suggests an intrinsic disease of cartilage in which biochemical and metabolic alterations result in its breakdown. Within the last decades different models were stablished which also concentrated on other joint structures such as bone or ligaments. Changes of the subchondral bone were found to precide cartilage damage suggesting a primary alteration of the subchondral region. Other studies concentrated on the metabolic activity of chondrocytes in healthy cartilage of osteoarthritis patients. The precise event that leads to these changes is still not clear. This review concentrates on the histological features in the course of the disease and tries to provide a summary on different pathogenetic concepts. (orig.) [de

  3. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain.

    Science.gov (United States)

    Hezel, Marcus; Ebrahimi, Fahim; Koch, Marco; Dehghani, Faramarz

    2012-10-01

    Immunohistochemical visualization of antigens in specimen has evolved to an indispensable technique in biomedical research for investigations of cell morphology and pathology both in bright field and fluorescence microscopy. While there are couple of staining methods that reveal entire cytoarchitecture in bright field microscopy such as Nissl or hemalaun-eosin, there are still limitations in visualizations of cytoarchitecture in fluorescence microscopy. The present study reports a simple staining method that provides the required illustration of cell allocations and cellular composition in fluorescence microscopy in adult and in developing rodent central nervous system using the fluorophore propidium iodide (PI, 5μg/mL). PI is a well-accepted marker for degenerating cells when applied prior to fixation (pre-fixation PI staining). Here, PI was added to the sections after the fixation (post-fixation PI staining). This revised labeling procedure led to similar cytoarchitectural staining patterns in fluorescence microscopy as observed with hemalaun in bright field microscopy. This finding was proven in organotypic hippocampal slice cultures (OHSC) and brain sections obtained from different postnatal developmental stages. Excitotoxically lesioned OHSC subjected to pre-fixation PI staining merely showed brightly labeled condensed nuclei of degenerating neurons. In contrast, post-fixation PI staining additionally revealed extensive labeling of neuronal cell bodies and glial cells within the OHSC, thus allowing visualization of stratification of neuronal layers and cell morphology. Furthermore, post-fixation PI staining was combined with NeuN, calbindin, calretinin, glial fibrillary acidic protein or Griffonia simplicifolia isolectin B4 (IB(4)) in post natal (p1 and p9) and adult rats. In early post-natal brain sections almost all mentioned cellular markers led to an incomplete staining of the native cell organization and resulted in an inaccurate estimation of cell

  4. Standardization in biological staining. The influence of dye manufacturing

    DEFF Research Database (Denmark)

    Lyon, H

    2000-01-01

    not have been subjected to quality assessment either internally by the producer or vendor or externally by independent investigators or organizations such as the Biological Stain Commission. Concerted attempts at standardization in Europe are discussed. The latest results of this work, the European...... standard EN 12376, is presented. This standard is concerned with information supplied by the manufacturer with in vitro diagnostic reagents for biological staining. The standard has been prepared by a Working Group on Staining in Biology under Technical Committee 140, In Vitro Medical Devices...

  5. Intraoperative immunohistochemistry staining of sentinel nodes in breast cancer: Clinical and economical implications

    DEFF Research Database (Denmark)

    Holm, M.; Paaschburg, B.; Balslev, E.

    2008-01-01

    .0001) for isolated tumor cell metastasis, from 56 to 36.4% (p analysis showed an overall...... of the surgical procedures before (n = 706) and after (n = 503) introducing intraoperatice IHC on frozen section. We also did a cost analysis. Intraoperative IHC staining led to a lowering of the late positive SNB rate. Introducing IHC gave a decrease in the late positive rate from 93 to 52% (p

  6. [Effects of surface roughness of bone cements on histological characteristics of induced membranes].

    Science.gov (United States)

    Liu, Hai-Xiao; Xu, Hua-Zi; Zhang, Yu; Hu, Gang; Shen, Yue; Cheng, Xiao-Jie; Peng, Lei

    2012-08-01

    To explore surface roughness of bone cement and surround tissue on histological characteristic of induced membranes. Bone cements with smooth and rough surface were implanted in radius bone defect, intramuscular and subcutaneous sites of rabbits, and formed induced membranes. Membranes were obtained and stained (HE) 6 weeks later. Images of membrane tissue were obtained and analyzed with an automated image analysis system. Five histological parameters of membranes were measured with thickness,area,cell density,ECM density and microvessel density. Double factor variance analysis was used to evaluate the effect of the two factors on histological characteristics of induced membranes. Membranes can be induced by each kind of bone cement and at all the three tissue sites. In histological parameters of thickness,area and micro vessel,there were significant differences among the membranes induced at different tissue sites (P = 0.000, P = 0.000, P = 0.000); whereas, there were no significant differences in histological parameters of cell density and ECM density (P = 0.734, P = 0.638). In all five histological parameters of membranes, there were no significant differences between the membranes induced by bone cements with different surface roughness (P = 0.506, P = 0.185, P = 0.883, P = 0.093, P = 0.918). Surround tissue rather than surface roughness of bone cements can affect the histological characteristics of induced membranes. The fibrocystic number, vascularity, mechanical tension and micro motion of the surround tissue may be closely correlated with the histological characteristics of induced membranes.

  7. The incidence of IgG4-positive plasma cells staining TIN in patients with biopsy-proven tubulointerstitial nephritis.

    Science.gov (United States)

    Mac, Kathy; Wu, Xiao Juan; Mai, Jun; Howlin, Kenneth; Suranyi, Michael; Yong, Jim; Makris, Angela

    2017-06-01

    IgG4 disease is rare. However, IgG4 tubulointerstitial nephritis (TIN) is the most common renal manifestation. IgG4 disease is usually associated with elevated serum IgG4 levels and other organ involvement, low-density renal lesions on enhanced CT imaging and immune activation. The incidence of IgG4-TIN may be underestimated, as staining for IgG4 is not routine. This study sought to describe the prevalence of previously undiagnosed IgG4-TIN. Due to the complexity of the diagnosis, we only attempt to look at IgG4-positive plasma cell TIN as a potential indication for IgG4 renal disease. A retrospective review of native renal biopsies performed between 2002 and 2012 with a primary diagnosis of TIN was selected. Samples for which interstitial nephritis was secondary to a glomerular disease were excluded. The tissues were stained for IgG4 and scored by two blinded observers. Demographic and follow-up details were collected. This study was approved by the local ethics committee. 82 cases of interstitial nephritis from a total of 1238 renal biopsies (2002-2012) were available after staining for further assessment. 12 samples demonstrated staining consistent with the criteria for IgG4-positive plasma cell TIN, of which 3 had mildly positive staining, 7 moderately positive staining and 2 had markedly positive staining. There were no statistically significant differences in the baseline characteristics between the positive and negative staining groups. A number of cases of IgG4-positive plasma cell TIN were observed histologically that had been previously diagnosed as non-specific chronic TIN. IgG4-positive plasma cell TIN made up 1% of all renal biopsies performed over 10 years and 13% of all biopsies demonstrating TIN not related to glomerular disease. IgG4 staining should be considered routinely in biopsies demonstrating primary TIN. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  8. MAL2 and tumor protein D52 (TPD52 are frequently overexpressed in ovarian carcinoma, but differentially associated with histological subtype and patient outcome

    Directory of Open Access Journals (Sweden)

    Fanayan Susan

    2010-09-01

    Full Text Available Abstract Background The four-transmembrane MAL2 protein is frequently overexpressed in breast carcinoma, and MAL2 overexpression is associated with gain of the corresponding locus at chromosome 8q24.12. Independent expression microarray studies predict MAL2 overexpression in ovarian carcinoma, but these had remained unconfirmed. MAL2 binds tumor protein D52 (TPD52, which is frequently overexpressed in ovarian carcinoma, but the clinical significance of MAL2 and TPD52 overexpression was unknown. Methods Immunohistochemical analyses of MAL2 and TPD52 expression were performed using tissue microarray sections including benign, borderline and malignant epithelial ovarian tumours. Inmmunohistochemical staining intensity and distribution was assessed both visually and digitally. Results MAL2 and TPD52 were significantly overexpressed in high-grade serous carcinomas compared with serous borderline tumours. MAL2 expression was highest in serous carcinomas relative to other histological subtypes, whereas TPD52 expression was highest in clear cell carcinomas. MAL2 expression was not related to patient survival, however high-level TPD52 staining was significantly associated with improved overall survival in patients with stage III serous ovarian carcinoma (log-rank test, p Conclusions MAL2 is frequently overexpressed in ovarian carcinoma, and TPD52 overexpression is a favourable independent prognostic marker of potential value in the management of ovarian carcinoma patients.

  9. The spinoglenoid ligament. Anatomy, morphology, and histological findings.

    Science.gov (United States)

    Plancher, Kevin D; Peterson, Robert K; Johnston, Jack C; Luke, Timothy A

    2005-02-01

    Dysfunction of the distal branch of the suprascapular nerve has been reported in athletes involved in throwing or overhead sports. The consistent presence of a dynamic anatomic structure, the spinoglenoid ligament, overlying the nerve in the spinoglenoid notch may be a contributing factor to the dysfunction of this nerve. The purpose of this study was to report the anatomy, morphology, and histological characteristics of the spinoglenoid ligament. The spinoglenoid ligaments of fifty-eight fresh-frozen cadaver shoulders were dissected to evaluate their anatomic dimensions, histological characteristics, and relationship to the suprascapular nerve, the posterior part of the capsule, and the glenoid rim. The spinoglenoid ligament was harvested, with its insertions on the scapular spine and on the capsule and glenoid left intact, for the histological analysis. Dissection revealed that a spinoglenoid ligament was present in all specimens. The ligament was found to form an irregular quadrangular shape. On gross examination, the deep fibers of the ligament extended from the lateral aspect of the scapular spine to the posterior part of the glenoid and the superficial fibers blended with the posterior aspect of the shoulder capsule. Histological sections demonstrated Sharpey fibers inserting into bone at the scapular spine and blending with the posterior aspect of the shoulder capsule to insert into the posterior surface of the glenoid, findings that confirmed the ligamentous nature of this structure. This study revealed the presence of the spinoglenoid ligament in all of the shoulders that were examined, with some variation in the size of the ligament.

  10. Histology of periapical lesions obtained during apical surgery.

    Science.gov (United States)

    Schulz, Malte; von Arx, Thomas; Altermatt, Hans Jörg; Bosshardt, Dieter

    2009-05-01

    The aim of this was to evaluate the histology of periapical lesions in teeth treated with periapical surgery. After root-end resection, the root tip was removed together with the periapical pathological tissue. Histologic sectioning was performed on calcified specimens embedded in methylmethacrylate (MMA) and on demineralized specimens embedded in LR White (Fluka, Buchs, Switzerland). The samples were evaluated with light and transmission electron microscopy (TEM). The histologic findings were classified into periapical abscesses, granulomas, or cystic lesions (true or pocket cysts). The final material comprised 70% granulomas, 23% cysts and 5% abscesses, 1% scar tissues, and 1% keratocysts. Six of 125 samples could not be used. The cystic lesions could not be subdivided into pocket or true cysts. All cysts had an epithelium-lined cavity, two of them with cilia-lined epithelium. These results show the high incidence of periapical granulomas among periapical lesions obtained during apical surgery. Periapical abscesses were a rare occasion. The histologic findings from samples obtained during apical surgery may differ from findings obtained by teeth extractions. A determination between pocket and true apical cysts is hardly possible when collecting samples by apical surgery.

  11. Immunohistochemical positive stained p53 protein in bladder transitional cell carcinoma

    Directory of Open Access Journals (Sweden)

    Halimi Monireh

    2009-04-01

    Full Text Available Background: Molecular genetics and immunopathologic analysis of bladder cancer have shown some abnormalities in a number of genes and proteins that have been implicated in the development and progression of such tumors, mainly in the p53 pathway. Aims: To investigate the rate of positively stained p53 protein in patients with urothelial papillary carcinoma of the bladder (UCB by immunohistochemistry and its relationship with tumor grade, gender and age of the patients. Settings and Design: During the present cross-sectional study, 100 paraffin-embedded specimens of UCB, which were provided from biopsies of the bladder by transurethral access, were immunohistochemically stained and studied for p53 protein from May 2006 to May 2007 in our referral center pathology laboratory. Materials and Methods: First, 4 µm slices of paraffin sections were provided and then stained by the avidin-biotin peroxidase method. The rate of positively stained p53 protein (defined as positive nuclear staining in over 10% of the cells was assessed. This rate was also estimated and compared between grades, genders and age-related groups (< 70 years, ≥70 years. Statistical Analysis: The χ2 , Fisher′s exact test and Mann-Whitney U test were used for comparing. Results: The overall rate of positively stained specimens was 11% for nuclear p53 protein. This rate was significantly higher in females (10/29 vs. 1/71; P < 0.001; odds ratio [OR]: 0.23; 95% confidence interval [CI]: 4.43-306.08, patients with 70 or older than 70 years (8/42 vs. 3/58; P = 0.04; OR: 0.55; 95% CI: 1.07-17.39 and in high-grade tumors (10/58 vs. 1/42; P = 0.02; OR: 0.59; 95% CI: 0.01-0.95. Conclusions: The rate of positively stained p53 protein for UCB was lower in our population. This rate was also higher in females, patients with 70 or older than 70 years and high grade of UCB.

  12. A Historical Perspective on the Identification of Cell Types in Pancreatic Islets of Langerhans by Staining and Histochemical Techniques.

    Science.gov (United States)

    Baskin, Denis G

    2015-08-01

    Before the middle of the previous century, cell types of the pancreatic islets of Langerhans were identified primarily on the basis of their color reactions with histological dyes. At that time, the chemical basis for the staining properties of islet cells in relation to the identity, chemistry and structure of their hormones was not fully understood. Nevertheless, the definitive islet cell types that secrete glucagon, insulin, and somatostatin (A, B, and D cells, respectively) could reliably be differentiated from each other with staining protocols that involved variations of one or more tinctorial techniques, such as the Mallory-Heidenhain azan trichrome, chromium hematoxylin and phloxine, aldehyde fuchsin, and silver impregnation methods, which were popularly used until supplanted by immunohistochemical techniques. Before antibody-based staining methods, the most bona fide histochemical techniques for the identification of islet B cells were based on the detection of sulfhydryl and disulfide groups of insulin. The application of the classical islet tinctorial staining methods for pathophysiological studies and physiological experiments was fundamental to our understanding of islet architecture and the physiological roles of A and B cells in glucose regulation and diabetes. © The Author(s) 2015.

  13. Standardization of fixation, processing and staining methods for the central nervous system of vertebrates.

    Science.gov (United States)

    Aldana Marcos, H J; Ferrari, C C; Benitez, I; Affanni, J M

    1996-12-01

    This paper reports the standardization of methods used for processing and embedding various vertebrate brains of different size in paraffin. Other technical details developed for avoiding frequent difficulties arising during laboratory routine are also reported. Some modifications of the Nissl and Klüver-Barrera staining methods are proposed. These modifications include: 1) a Nissl stain solution with a rapid and efficient action with easier differentiation; 2) the use of a cheap microwave oven for the Klüver-Barrera stain. These procedures have the advantage of permitting Nissl and Klüver-Barrera staining of nervous tissue in about five and fifteen minutes respectively. The proposed procedures have been tested in brains obtained from fish, amphibians, reptiles and mammals of different body sizes. They are the result of our long experience in preparing slides for comparative studies. Serial sections of excellent quality were regularly obtained in all the specimens studied. These standardized methods, being simple and quick, are recommended for routine use in neurobiological laboratories.

  14. Productivity standards for histology laboratories.

    Science.gov (United States)

    Buesa, René J

    2010-04-01

    The information from 221 US histology laboratories (histolabs) and 104 from 24 other countries with workloads from 600 to 116 000 cases per year was used to calculate productivity standards for 23 technical and 27 nontechnical tasks and for 4 types of work flow indicators. The sample includes 254 human, 40 forensic, and 31 veterinary pathology services. Statistical analyses demonstrate that most productivity standards are not different between services or worldwide. The total workload for the US human pathology histolabs averaged 26 061 cases per year, with 54% between 10 000 and less than 30 000. The total workload for 70% of the histolabs from other countries was less than 20 000, with an average of 15 226 cases per year. The fundamental manual technical tasks in the histolab and their productivity standards are as follows: grossing (14 cases per hour), cassetting (54 cassettes per hour), embedding (50 blocks per hour), and cutting (24 blocks per hour). All the other tasks, each with their own productivity standards, can be completed by auxiliary staff or using automatic instruments. Depending on the level of automation of the histolab, all the tasks derived from a workload of 25 cases will require 15.8 to 17.7 hours of work completed by 2.4 to 2.7 employees with 18% of their working time not directly dedicated to the production of diagnostic slides. This article explains how to extrapolate this productivity calculation for any workload and different levels of automation. The overall performance standard for all the tasks, including 8 hours for automated tissue processing, is 3.2 to 3.5 blocks per hour; and its best indicator is the value of the gross work flow productivity that is essentially dependent on how the work is organized. This article also includes productivity standards for forensic and veterinary histolabs, but the staffing benchmarks for histolabs will be the subject of a separate article. Copyright 2010 Elsevier Inc. All rights reserved.

  15. Histological characteristics of diffuse idiopathic skeletal hyperostosis

    NARCIS (Netherlands)

    Kuperus, JS; Westerveld, L Anneloes; Rutges, Joost A; Alblas, Jacqueline; van Rijen, Mattie H; Bleys, Ronald L A W; Oner, F Cumhur; Verlaan, JJ

    Diffuse idiopathic skeletal hyperostosis (DISH) is a predominantly radiographic diagnosis and histological knowledge of DISH is limited. The aim of this study was to describe the histological characteristics of DISH in the spinal column and to study the relation between DISH and intervertebral disc

  16. Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine.

    Science.gov (United States)

    Thomas, Marlon S; Nuñez, Vicente; Upadhyayula, Srigokul; Zielins, Elizabeth R; Bao, Duoduo; Vasquez, Jacob M; Bahmani, Baharak; Vullev, Valentine I

    2010-06-15

    For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.

  17. Microscopic analysis of MTT stained boar sperm cells

    African Journals Online (AJOL)

    tulyasys

    2015-06-08

    2H-tetrazolium bromide is widely used for assessment of cytotoxicity, cell viability, and proliferation studies in cell biology (van Meerloo et al., 2011;. Stockert et al., 2012). The stain is abbreviated as MTT.

  18. Color model comparative analysis for breast cancer diagnosis using H and E stained images

    Science.gov (United States)

    Li, Xingyu; Plataniotis, Konstantinos N.

    2015-03-01

    Digital cancer diagnosis is a research realm where signal processing techniques are used to analyze and to classify color histopathology images. Different from grayscale image analysis of magnetic resonance imaging or X-ray, colors in histopathology images convey large amount of histological information and thus play significant role in cancer diagnosis. Though color information is widely used in histopathology works, as today, there is few study on color model selections for feature extraction in cancer diagnosis schemes. This paper addresses the problem of color space selection for digital cancer classification using H and E stained images, and investigates the effectiveness of various color models (RGB, HSV, CIE L*a*b*, and stain-dependent H and E decomposition model) in breast cancer diagnosis. Particularly, we build a diagnosis framework as a comparison benchmark and take specific concerns of medical decision systems into account in evaluation. The evaluation methodologies include feature discriminate power evaluation and final diagnosis performance comparison. Experimentation on a publicly accessible histopathology image set suggests that the H and E decomposition model outperforms other assessed color spaces. For reasons behind various performance of color spaces, our analysis via mutual information estimation demonstrates that color components in the H and E model are less dependent, and thus most feature discriminate power is collected in one channel instead of spreading out among channels in other color spaces.

  19. Analysis of prevalence and clinical features of multicystic ameloblastoma and its histological subtypes in South Indian sample population: A retrospective study over 13 years

    Directory of Open Access Journals (Sweden)

    Manickam Selvamani

    2014-01-01

    Full Text Available Objective: The study was designed to analyze the frequency and clinical features of multicystic ameloblastoma and its histological variants in South Indian sample population, as there is minute information available in the English literature. Methodology: The study source was the biopsy specimens retrieved from the archives of the Department of Oral and Maxillofacial Pathology, College of Dental Sciences, Davangere, Karnataka, India, during the past 13 years, from 2001 to 2013. Clinical data for the study were obtained from the case records of patients and the analyzed clinical variables were age, gender and anatomical location. Histologically, hematoxylin and eosin stained sections fitting the World Health Organization (2005 criteria for diagnosis of multicystic ameloblastoma were selectively included. Results: Of the 3026 biopsy reports analyzed, 103 cases were odontogenic tumors (3.4% and 58 cases were ameloblastoma. 31 cases of multicystic ameloblastoma, including follicular ameloblastoma (54.8%, acanthomatous ameloblastoma (29%, plexiform ameloblastoma (6.5%, granular cell ameloblastoma (6.5% and desmoplastic ameloblastoma (3.2% were recorded. The age of the patients during the presentation of the lesion was ranging from 21 to 73 years, with a mean of 39.5 years. The most frequent clinical manifestation was swelling, followed by a combination of pain and swelling. In our study, ameloblastoma showed distinct anatomic predilections for occurrence in mandible (96.8% rather than maxilla (3.2%. This study result also indicated that there is geographical variation in the frequency and distribution of ameloblastoma.

  20. A new rapid immunohistochemical staining technique using the EnVision antibody complex.

    Science.gov (United States)

    Kämmerer, U; Kapp, M; Gassel, A M; Richter, T; Tank, C; Dietl, J; Ruck, P

    2001-05-01

    Rapid immunohistochemical investigation, in addition to staining with hematoxylin and eosin, would be useful during intraoperative frozen section diagnosis in some cases. This study was undertaken to investigate whether the recently described EnVision system, a highly sensitive two-step immunohistochemical technique, could be modified for rapid immunostaining of frozen sections. Forty-five primary antibodies were tested on frozen sections from various different tissues. After fixation in acetone for 1 min and air-drying, the sections were incubated for 3 min each with the primary antibody, the EnVision complex (a large number of secondary antibodies and horseradish peroxidase coupled to a dextran backbone), and the chromogen (3,3'diaminobenzidine or 3-amino-9-ethylcarbazole). All reactions were carried out at 37C. Specific staining was seen with 38 antibodies (including HMB-45 and antibodies against keratin, vimentin, leukocyte common antigen, smooth muscle actin, synaptophysin, CD34, CD3, CD20, and prostate-specific antigen). A modification of the EnVision method allows the detection of a broad spectrum of antigens in frozen sections in less than 13 min. This method could be a useful new tool in frozen section diagnosis and research. (J Histochem Cytochem 49:623-630, 2001)

  1. Gram staining for the treatment of peritonsillar abscess.

    Science.gov (United States)

    Takenaka, Yukinori; Takeda, Kazuya; Yoshii, Tadashi; Hashimoto, Michiko; Inohara, Hidenori

    2012-01-01

    Objective. To examine whether Gram staining can influence the choice of antibiotic for the treatment of peritonsillar abscess. Methods. Between 2005 and 2009, a total of 57 cases of peritonsillar abscess were analyzed with regard to cultured bacteria and Gram staining. Results. Only aerobes were cultured in 16% of cases, and only anaerobes were cultured in 51% of cases. Mixed growth of aerobes and anaerobes was observed in 21% of cases. The cultured bacteria were mainly aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. Phagocytosis of bacteria on Gram staining was observed in 9 cases. The bacteria cultured from these cases were aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. The sensitivity of Gram staining for the Gram-positive cocci and Gram-negative rods was 90% and 64%, respectively. The specificity of Gram staining for the Gram-positive cocci and Gram-negative rods was 62% and 76%, respectively. Most of the Gram-positive cocci were sensitive to penicillin, but some of anaerobic Gram-negative rods were resistant to penicillin. Conclusion. When Gram staining shows only Gram-positive cocci, penicillin is the treatment of choice. In other cases, antibiotics effective for the penicillin-resistant organisms should be used.

  2. An improved method for staining cell colonies in clonogenic assays.

    Science.gov (United States)

    Guda, Kishore; Natale, Leanna; Markowitz, Sanford D

    2007-06-01

    Clonogenic assay is a widely used experimental approach to test for the effects of drugs/genes on the growth and proliferative characteristics of cells in vitro. Accurate quantitation of treatment effects in clonogeneic assays depends on the ability to visualize and count cell colonies precisely. We report a novel method (referred as ETeB) for staining cell colonies grown on plastic and specially coated substrates like collagen. Using colon cancer cell lines grown on plastic and collagen, we compared the colony staining efficiencies of the widely used methylene blue, and Ethidium bromide (ETeB) stains. Results show that the ETeB protocol works well on plastic and is extremely effective for staining colonies on collagen when compared to methylene blue. The key features and advantages of ETeB technique are; (a) reduction in background for colonies grown on collagen and possibly other substrates, (b) the whole procedure takes less than a minute, (c) no post-stain washing step is required which eliminates colony losses for cell lines that are loosely adherent, (d) colony visualization and counting can be done immediately following the staining procedure using a standard UV illuminator and software, and (e) the method works across a wide variety of cell lines. The simplicity and robustness of this procedure should warrant its usage in both small and large-scale clonogenic experiments.

  3. MANAJEMEN SARANA DAN PRASARANA PENDIDIKAN DI STAIN PAMEKASAN

    Directory of Open Access Journals (Sweden)

    M. Muchlis Solichin

    2011-07-01

    Full Text Available Mediums management and pre-mediums represent an absolute done in an higher education institute, because Mediums and premediums in education management represent the absolut condition in the effort to reach the target which is expected. Thereby, Every the education organizer have to pay attention and conscripting the mind and energy to carry out education management that is professional and fulfill Standard National Education ( SNP. This Research copes to comprehend the mediums and pre-mediums management of education in STAIN Pamekasan, because during this time of mediums and basic mediums management are not yet showing its idealitas. This research is focussed at; a How mediums and pre-mediums menegement in STAIN Pamekasan ?,and b what Factors influencing mediums and pre-mediums management in STAIN Pamekasan ?. This research uses the qualitative type by using observation, interview, and documentation method. Based the rearch done, to be expressed that the first of STAIN Pamekasan conduct mediums and pre-mediums manegement still have the centralization character of top down, either in the case of planning, organizational, observation, and assessment of mediums and pre-mediums management owned, second in some cases of STAIN Pamekasan do not yet manage the mediums and pre-mediums management because they are caused by factor is its lack of management professionalism, either when doing the planning, organizational, treatment and observation or evaluation. Based the matter above, hence, suggested that STAIN Pamekasan carry out the mediums and pre-mediums management of education professionally.

  4. An optimized color transformation for the analysis of digital images of hematoxylin & eosin stained slides

    Directory of Open Access Journals (Sweden)

    Mark D Zarella

    2015-01-01

    Full Text Available Hematoxylin and eosin (H&E staining is ubiquitous in pathology practice and research. As digital pathology has evolved, the reliance of quantitative methods that make use of H&E images has similarly expanded. For example, cell counting and nuclear morphometry rely on the accurate demarcation of nuclei from other structures and each other. One of the major obstacles to quantitative analysis of H&E images is the high degree of variability observed between different samples and different laboratories. In an effort to characterize this variability, as well as to provide a substrate that can potentially mitigate this factor in quantitative image analysis, we developed a technique to project H&E images into an optimized space more appropriate for many image analysis procedures. We used a decision tree-based support vector machine learning algorithm to classify 44 H&E stained whole slide images of resected breast tumors according to the histological structures that are present. This procedure takes an H&E image as an input and produces a classification map of the image that predicts the likelihood of a pixel belonging to any one of a set of user-defined structures (e.g., cytoplasm, stroma. By reducing these maps into their constituent pixels in color space, an optimal reference vector is obtained for each structure, which identifies the color attributes that maximally distinguish one structure from other elements in the image. We show that tissue structures can be identified using this semi-automated technique. By comparing structure centroids across different images, we obtained a quantitative depiction of H&E variability for each structure. This measurement can potentially be utilized in the laboratory to help calibrate daily staining or identify troublesome slides. Moreover, by aligning reference vectors derived from this technique, images can be transformed in a way that standardizes their color properties and makes them more amenable to image

  5. An optimized color transformation for the analysis of digital images of hematoxylin & eosin stained slides.

    Science.gov (United States)

    Zarella, Mark D; Breen, David E; Plagov, Andrei; Garcia, Fernando U

    2015-01-01

    Hematoxylin and eosin (H&E) staining is ubiquitous in pathology practice and research. As digital pathology has evolved, the reliance of quantitative methods that make use of H&E images has similarly expanded. For example, cell counting and nuclear morphometry rely on the accurate demarcation of nuclei from other structures and each other. One of the major obstacles to quantitative analysis of H&E images is the high degree of variability observed between different samples and different laboratories. In an effort to characterize this variability, as well as to provide a substrate that can potentially mitigate this factor in quantitative image analysis, we developed a technique to project H&E images into an optimized space more appropriate for many image analysis procedures. We used a decision tree-based support vector machine learning algorithm to classify 44 H&E stained whole slide images of resected breast tumors according to the histological structures that are present. This procedure takes an H&E image as an input and produces a classification map of the image that predicts the likelihood of a pixel belonging to any one of a set of user-defined structures (e.g., cytoplasm, stroma). By reducing these maps into their constituent pixels in color space, an optimal reference vector is obtained for each structure, which identifies the color attributes that maximally distinguish one structure from other elements in the image. We show that tissue structures can be identified using this semi-automated technique. By comparing structure centroids across different images, we obtained a quantitative depiction of H&E variability for each structure. This measurement can potentially be utilized in the laboratory to help calibrate daily staining or identify troublesome slides. Moreover, by aligning reference vectors derived from this technique, images can be transformed in a way that standardizes their color properties and makes them more amenable to image processing.

  6. Digital transplantation pathology: combining whole slide imaging, multiplex staining and automated image analysis.

    Science.gov (United States)

    Isse, K; Lesniak, A; Grama, K; Roysam, B; Minervini, M I; Demetris, A J

    2012-01-01

    Conventional histopathology is the gold standard for allograft monitoring, but its value proposition is increasingly questioned. "-Omics" analysis of tissues, peripheral blood and fluids and targeted serologic studies provide mechanistic insights into allograft injury not currently provided by conventional histology. Microscopic biopsy analysis, however, provides valuable and unique information: (a) spatial-temporal relationships; (b) rare events/cells; (c) complex structural context; and (d) integration into a "systems" model. Nevertheless, except for immunostaining, no transformative advancements have "modernized" routine microscopy in over 100 years. Pathologists now team with hardware and software engineers to exploit remarkable developments in digital imaging, nanoparticle multiplex staining, and computational image analysis software to bridge the traditional histology-global "-omic" analyses gap. Included are side-by-side comparisons, objective biopsy finding quantification, multiplexing, automated image analysis, and electronic data and resource sharing. Current utilization for teaching, quality assurance, conferencing, consultations, research and clinical trials is evolving toward implementation for low-volume, high-complexity clinical services like transplantation pathology. Cost, complexities of implementation, fluid/evolving standards, and unsettled medical/legal and regulatory issues remain as challenges. Regardless, challenges will be overcome and these technologies will enable transplant pathologists to increase information extraction from tissue specimens and contribute to cross-platform biomarker discovery for improved outcomes. ©Copyright 2011 The American Society of Transplantation and the American Society of Transplant Surgeons.

  7. Diffusion tensor microscopy data (15.6 μm in-plane of white matter tracts in the human, pig, and rat spinal cord with corresponding tissue histology

    Directory of Open Access Journals (Sweden)

    Jeremy J. Flint

    2016-12-01

    Full Text Available The following article contains nine diffusion tensor imaging (DTI datasets acquired with magnetic resonance microscopy (MRM, 15.6 μm in-plane. All data was collected in the region bordering the ventral horn and white matter of cross sections from the spinal cord enlargements along with each sample׳s corresponding tissue histology. These data are collected in fixed spinal cord sections of varying thicknesses taken from rat (2×21 direction DTI datasets, pig (1×21 direction DTI dataset, and human (5×21 direction DTI datasets + 1×6 direction DTI dataset tissue sources. Following MRM acquisition, the sections were histologically processed using Nissl or Black-Gold II (Histo-Chem Inc., 1BGII myelin stain and imaged again using light microscopy techniques. Methodological procedures are an amalgamation of protocol components described previously (doi:10.1016/j.neuroimage.2010.04.031 [1], doi:10.1016/j.neuroimage.2011.04.052 [2].

  8. Biomechanical and histological effects of augmented soft tissue mobilization therapy on achilles tendinopathy in a rabbit model.

    Science.gov (United States)

    Imai, Kan; Ikoma, Kazuya; Chen, Qingshan; Zhao, Chunfeng; An, Kai-Nan; Gay, Ralph E

    2015-02-01

    Augmented soft tissue mobilization (ASTM) has been used to treat Achilles tendinopathy and is thought to promote collagen fiber realignment and hasten tendon regeneration. The objective of this study was to evaluate the biomechanical and histological effects of ASTM therapy on rabbit Achilles tendons after enzymatically induced injury. This study was a non-human bench controlled research study using a rabbit model. Both Achilles tendons of 12 rabbits were injected with collagenase to produce tendon injury simulating Achilles tendinopathy. One side was then randomly allocated to receive ASTM, while the other received no treatment (control). ASTM was performed on the Achilles tendon on postoperative days 21, 24, 28, 31, 35, and 38. Tendons were harvested 10 days after treatment and examined with dynamic viscoelasticity and light microscopy. Cross-sectional area in the treated tendons was significantly greater than in controls. Storage modulus tended to be lower in the treated tendons but elasticity was not significantly increased. Loss modulus was significantly lower in the treated tendons. There was no significant difference found in tangent delta (loss modulus/storage modulus). Microscopy of control tendons showed that the tendon fibers were wavy and type III collagen was well stained. The tendon fibers of the augmented soft tissue mobilization treated tendons were not wavy and type III collagen was not prevalent. Biomechanical and histological findings showed that the Achilles tendons treated with ASTM had better recovery of biomechanical function than did control tendons. Copyright © 2015 National University of Health Sciences. Published by Elsevier Inc. All rights reserved.

  9. Endometrial Histology of Depomedroxyprogesterone Acetate Users: A Pilot Study

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available Objective. To obtain pilot data on the endometrial histology of Depomedroxyprogesterone acetate (Depo-Provera, DMPA users experiencing breakthrough bleeding (BTB versus users with amenorrhea. To compare the endometrial histology of patients who used DMPA continuously for 3–12 months versus those who used it for 13 months or more. Methods. Cross-sectional study. Endometrial biopsy was obtained from all consenting patients who used DMPA for at least 3 months. Patients were divided into those with BTB in the last 3 months versus those with amenorrhea for at least 3 months. Histology results and duration of therapy were compared. Results. The proportion of women with chronic endometritis, uterine polyps, atrophic, proliferative, or progesterone-dominant endometrium did not differ between those DMPA users with BTB versus those with amenorrhea. Duration of therapy did not correlate with symptoms of BTB or endometrial histology. Chronic endometritis was the most common histologic finding (10/40, 25% and occurred more often in women experiencing BTB (35% versus 15% (RR 1.62 CI 0.91–2.87. Moreover, 45% of women with BTB had received DMPA for more than 12 months. Conclusions. BTB was more common than previously reported in women using DMPA for more than 12 months. Chronic endometritis, which may indicate an underlying infectious or intracavitary anatomic etiology, has not been previously reported as a frequent finding in DMPA users, and may be related to ethnic or other sociodemographic characteristics of our patient population. Further study to elucidate the etiology of chronic endometritis in these patients is warranted.

  10. Gloss and Stain Resistance of Ceramic-Polymer CAD/CAM Restorative Blocks.

    Science.gov (United States)

    Lawson, Nathaniel C; Burgess, John O

    2016-03-01

    To evaluate the gloss and stain resistance of several new ceramic-polymer CAD/CAM blocks Specimens (4 mm) were sectioned from: Enamic (polymer-infused ceramic), LAVA Ultimate (nano-ceramic reinforced polymer), e.max (lithium disilicate), Paradigm C (porcelain), and Paradigm MZ100 (composite). Specimens were wet polished on a polishing wheel to either 320 grit silicon paper (un-polished, N = 8) or 2000 grit silicon carbide papers followed by a 0.05 μm alumina slurry (polished, N = 8). Initial gloss and color (L*a*b*) values were measured. Specimens were stored in a staining solution at 37°C in darkness for 12 days (simulating 1 year). After storage, L*a*b* values re-measured. Change in color was reported as ΔE00 based on the CIEDE2000 formula. Gloss and ΔE00 were analyzed by two-way analysis of variance (ANOVA) (alpha = .05). Separate one-way ANOVA and Tukey post-hoc analyses were performed for both polish conditions and all materials. Two-way ANOVA showed that factors material, polish and their interaction were significant for both gloss and ΔE00 (p gloss and less color change than all other materials. The composition and polish of CAD/CAM materials affects gloss and stain resistance. Ceramic-polymer hybrid materials can achieve the high gloss required for esthetic restorations. These materials should be polished in order to minimize staining. If polished, all of the tested materials exhibited clinically acceptable color changes at 1 year of simulated staining. (J Esthet Restor Dent 28:S40-S45, 2016). © 2015 Wiley Periodicals, Inc.

  11. QUANTITATIVE STUDY OF GASTRIC EPITHELIAL LESIONS BY NUCLEOLAR ORGANIZER REGION STAINING

    Directory of Open Access Journals (Sweden)

    M.R. Arab

    2004-11-01

    Full Text Available Nucleolar organizer regions (NOR are defined as nucleolar components containing a set of argyrophilic proteins which are selectively stained by colloidal silver nitrate staining. Although studies have shown that the number of NOR dots or particles is directly related to the rapidity of cell proliferation in cancer cells, prognostic or diagnostic value of NOR remains controversial. The aim of the present study was to asses the proliferative activity of the NOR in different gastric epithelial lesions. For these purposes 60 biopsy and surgical specimens of stomach from pathology files of Khatamalanbia and Imam Hospitals were chosen. For each patient, 3-5 paraffin sections were prepared and stained by one step colloidal silver nitrate solution. In each section intranuclear dots in 100 cell nuclei were counted by two of authors in randomly selected fields and data were analyzed by ANOVA. Statistical analysis showed significant difference for NOR number between gastritis, different grades of dysplasia and carcinoma. The shape and number of NOR showed a grater variability in carcinoma compared to other lesions. It seems that NOR could reflect the proliferative activity of cells.

  12. Histology study of the somatic embryogenesis in Agave fourcroydes Lem.

    Directory of Open Access Journals (Sweden)

    Silvia Alemán García

    2002-01-01

    Full Text Available A histological study of the structures embryogenic formed from meristem of Agave fourcroydes, with the objective of observing the behavior of the somatic embryogenesis from the induction until the formation of the embryo was carried out. The meristem cultivated in vitro was subculture in modified MS medium the induction. The samples to be included intheresinJB–4weretakenperiodically.Thehistologicalscutsof5-10µmwerecarriedoutwithrotatorymicrotome and strained in 2% toluidine blue. The sections were observed and photographed in light microscopy Axioplan Zeiis with increases of 20X. The results show to the 60 days of culture the presence of cells meristematic pre-embrygenic. Embryos in different phases of formation globular and scutellum were observed in all the mass of the callus in the following subculture. Key words: Agavaceae, in vitro culture, embryo, histology

  13. Quantification of histochemical stains using whole slide imaging: development of a method and demonstration of its usefulness in laboratory quality control.

    Science.gov (United States)

    Gray, Allan; Wright, Alex; Jackson, Pete; Hale, Mike; Treanor, Darren

    2015-03-01

    Histochemical staining of tissue is a fundamental technique in tissue diagnosis and research, but it suffers from significant variability. Efforts to address this include laboratory quality controls and quality assurance schemes, but these rely on subjective interpretation of stain quality, are laborious and have low reproducibility. We aimed (1) to develop a method for histochemical stain quantification using whole slide imaging and image analysis and (2) to demonstrate its usefulness in measuring staining variation. A method to quantify the individual stain components of histochemical stains on virtual slides was developed. It was evaluated for repeatability and reproducibility, then applied to control sections of an appendix to quantify H&E staining (H/E intensities and H:E ratio) between automated staining machines and to measure differences between six regional diagnostic laboratories. The method was validated with laboratories from 0.57 to 0.89. A simple method using whole slide imaging can be used to quantify and compare histochemical staining. This method could be deployed in routine quality assurance and quality control. Work is needed on whole slide imaging devices to improve reproducibility. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  14. Staining for factor VIII related antigen and Ulex europaeus agglutinin I (UEA-I) in 230 tumours. An assessment of their specificity for angiosarcoma and Kaposi's sarcoma.

    Science.gov (United States)

    Leader, M; Collins, M; Patel, J; Henry, K

    1986-11-01

    In this study we examined the staining reactivity of commercially available antisera to factor VIII related antigen (F VIII RAg) and Ulex europaeus agglutinin I (UEA-I) on sections from 230 formalin fixed paraffin embedded tumours. These included 196 sarcomas, 20 carcinomas and 14 angiomas. All angiomas showed positive staining for F VIII RAg; all carcinomas showed negative staining; the vasoformative areas of all angiosarcomas stained positively but only four of six angiosarcomas showed positive staining of their solid areas; of seven Kaposi's sarcomas, all showed positive staining of vessels and six showed positive staining of the spindle cell component. In the remaining 181 non-vascular sarcomas there was a false positive result in four tumours (2.2%), three of which had a history of irradiation. Pre-radiotherapy biopsies of these three tumours stained negatively with anti-F VIII RAg. UEA-I was demonstrated in all the angiomas studied, in all angiosarcomas (including the solid components) and in well-formed vessels of all Kaposi's sarcomas, but only in the spindle cell component of 3/6. However, there was an unacceptably high rate of false positive staining amongst the carcinomas and non-vascular sarcomas. In conclusion, F VIII RAg is a specific but not a sensitive marker of angiosarcomas; UEA-I is a sensitive but not a specific marker of angiosarcomas.

  15. Association of Diet With Skin Histological Features in UV-B-Exposed Mice.

    Science.gov (United States)

    Bhattacharyya, Tapan K; Hsia, Yvonne; Weeks, David M; Dixon, Tatiana K; Lepe, Jessica; Thomas, J Regan

    2017-09-01

    Long-term exposure to solar radiation produces deleterious photoaging of the skin. It is not known if diet can influence skin photoaging. To study the influence of a calorie-restricted diet and an obesity diet in mice exposed to long-term UV-B irradiation to assess if there is an association between diet and histopathological response to UV-B irradiation. In this animal model study in an academic setting, the dorsal skin of SKH1 hairless mice receiving normal, calorie-restricted, and obesity diets was exposed to UV-B irradiation 3 times a week for 10 weeks and were compared with corresponding controls. The mice were placed in the following groups, with 8 animals in each group: (1) intact control (C) with regular diet and no UV-B exposure, (2) intact control with UV-B exposure (CR), (3) calorie-restricted diet (CrC), (4) calorie-restricted diet with UV-B exposure (CrR), (5) obesity diet (OC), and (6) obesity diet with UV-B exposure (OR). The experiment was conducted during October through December 2013. Tissue processing and histological analysis were completed in 2016. Histomorphometric analysis was performed on paraffin-embedded skin sections stained by histological and immunohistochemical methods for estimation of epidermal thickness, epidermal proliferating cell nuclear antigen index, collagen I, elastic fibers, fibroblasts, mast cells, dermal cellularity, and adipose layer ratio. Changes in wrinkles were noted. Hairless female mice (age range, 6-8 weeks) were obtained. With a normal diet, changes from UV-B irradiation occurred in epidermal thickness, epidermal proliferating cell nuclear antigen index, collagen I, elastic fibers, fibroblasts, and mast cells, which were modestly influenced by an obesity diet. Calorie restriction influenced the skin in nonirradiated control animals, with higher values for most variables. After UV-B exposure in animals with calorie restriction, epidermal thickness was increased, but other variables were unaffected. Animals

  16. A histological procedure to study fungal infection in the wax moth Galleria mellonella

    Directory of Open Access Journals (Sweden)

    F. Perdoni

    2014-09-01

    Full Text Available The invertebrate model Galleria mellonella is a widely used factitious host to study the microbial pathogenesis in vivo. However, a specific procedure for the recovery and the processing of the infected tissues, important for a better understanding of the host-pathogen interactions, has not been reported to our knowledge. In the present study we describe a new procedure of fixation and processing of larval tissue that allows studying the larval topographic anatomy and assessing the morphological changes due to the fungal infection. Lepidopteran larvae were infected with Candida albicans strains displaying various biofilm-forming abilities. The whole larvae were then examined for tissue changes by histological techniques. We show that comparing cutting planes, serial transversal sections of paraffin-embedded larva result in better accuracy and information recovering. Using this technique, it was possible to preserve the integrity of G. mellonella internal structures allowing the detailed analysis of morphological differences in different experimental groups (i.e., healthy vs infected larvae. We were also able to study strain-related differences in the pathogenesis of C. albicans by observing the immune response elicited and the invasiveness of two isolates within the larval tissues. In general, by processing the whole larva and optimizing routinely histochemical stainings, it is possible to visualize and analyse infected tissues. Various degrees of pathogenicity (strain- or inoculum-related, and the infection time course can be described in details. Moreover, the host immune response events can be followed throughout the infectious process leading to a comprehensive picture of the studied phenomenon.

  17. Segmentation of histological images and fibrosis identification with a convolutional neural network.

    Science.gov (United States)

    Fu, Xiaohang; Liu, Tong; Xiong, Zhaohan; Smaill, Bruce H; Stiles, Martin K; Zhao, Jichao

    2018-05-16

    Segmentation of histological images is one of the most crucial tasks for many biomedical analyses involving quantification of certain tissue types, such as fibrosis via Masson's trichrome staining. However, challenges are posed by the high variability and complexity of structural features in such images, in addition to imaging artifacts. Further, the conventional approach of manual thresholding is labor-intensive, and highly sensitive to inter- and intra-image intensity variations. An accurate and robust automated segmentation method is of high interest. We propose and evaluate an elegant convolutional neural network (CNN) designed for segmentation of histological images, particularly those with Masson's trichrome stain. The network comprises 11 successive convolutional - rectified linear unit - batch normalization layers. It outperformed state-of-the-art CNNs on a dataset of cardiac histological images (labeling fibrosis, myocytes, and background) with a Dice similarity coefficient of 0.947. With 100 times fewer (only 300,000) trainable parameters than the state-of-the-art, our CNN is less susceptible to overfitting, and is efficient. Additionally, it retains image resolution from input to output, captures fine-grained details, and can be trained end-to-end smoothly. To the best of our knowledge, this is the first deep CNN tailored to the problem of concern, and may potentially be extended to solve similar segmentation tasks to facilitate investigations into pathology and clinical treatment. Copyright © 2018. Published by Elsevier Ltd.

  18. Validation of optical coherence tomography in vivo using cryostat histology

    International Nuclear Information System (INIS)

    Gambichler, T; Moussa, G; Regeniter, P; Kasseck, C; Hofmann, M R; Bechara, F G; Sand, M; Altmeyer, P; Hoffmann, K

    2007-01-01

    We aimed to validate for the first time optical coherence tomography (OCT) measurements of epidermal thickness (ET) using cryopreparation for histology. OCT assessments of ET were performed on healthy skin using the algorithms as follows: first, peak-to-valley analysis of the A-scan (ET-OCT-V), second, line-traced image analysis of the B-scan (ET-OCT-IA). Histology was performed using cryostat sections which were also evaluated using the image analysis (ET-Histo). We selected 114 samples, including B-scans and corresponding histology, for method comparison between ET-OCT-IA and ET-Histo. Forty-two A-scans were available for method comparison between ET-OCT-V and ET-Histo. Bland and Altman plots revealed a marked bias with wide 95% limits of agreement for ET-OCT-V versus ET-Histo. Comparison of ET-OCT-IA versus ET-Histo revealed only a slight bias and narrow 95% limits of agreement. A-scan analysis for ET determination is linked to significant limitations and lacks agreement with histology. By contrast, we observed satisfactory agreement between ET-OCT-IA and ET-Histo indicating that both methods can be utilized interchangeably. OCT using the line-traced image analysis of the B-scan appears to be a valid and relatively practicable method for the determination of ET in vivo. Furthermore, the comparisons with the in vivo OCT profiles demonstrate that cryostat sectioning provides a better preservation of relative and absolute dimensions of skin layers than paraffin embedding. (note)

  19. Intrapartum amnioinfusion in meconium-stained liquor: a case-control study.

    Science.gov (United States)

    Bansal, Neeta; Gupta, Vineeta; Nanda, Anuja; Chaudhary, Priyanka; Tandon, Archna; Behl, Neelima

    2013-06-01

    The aim of this study was to investigate perinatal outcome and the rate of cesarean section (CS) following intrapartum amnioinfusion in women with meconium-stained amniotic fluid (MSAF). A total of 100 women at term in labor with meconium were randomized to infuse transcervical intrapartum amnioinfusion with saline (50) and routine obstetrical care (50). Perinatal outcome and obstetric outcome were recorded and analyzed in both groups by means of Chi-square test. The CS rate due to fetal distress was 40.0 % in the control group and 20.0 % in the study group. The difference was statistically significant (P Amnioinfusion in cases of meconium-stained liquor significantly improved neonatal outcome and CS rate without increasing any maternal and fetal complications.

  20. Red layered medieval stained glass window characterization by means of micro-PIXE technique

    Energy Technology Data Exchange (ETDEWEB)

    Ortega-Feliu, I., E-mail: iofeliu@us.es [Centro Nacional de Aceleradores, Universidad de Sevilla, Avda. Thomas A. Edison 7, 41092 Sevilla (Spain); Gomez-Tubio, B. [Centro Nacional de Aceleradores, Universidad de Sevilla, Avda. Thomas A. Edison 7, 41092 Sevilla (Spain); Departamento de Fisica Aplicada III, Universidad de Sevilla (Spain); Respaldiza, M.A. [Centro Nacional de Aceleradores, Universidad de Sevilla, Avda. Thomas A. Edison 7, 41092 Sevilla (Spain); Departamento de Fisica Atomica, Molecular y Nuclear, Universidad de Sevilla (Spain); Capel, F. [Instituto de Ceramica y Vidrio, Consejo Superior de Investigaciones Cientificas (Spain)

    2011-10-15

    Red layered medieval stained glass windows on a transparent greenish substrate are characteristic of European medieval cathedrals, but few compositional analyses have been performed on the coloured layers. The PIXE technique has been performed on a red layered stained glass window obtained during the restoration works carried out in Las Huelgas Monastery in Burgos (Spain). Protons of 3 MeV with a beam of 4 x 5 {mu}m{sup 2} were used to acquire elemental maps of a cross section of the sample, in order to observe the homogeneity of the layered structure and its substrate. In our work, copper was detected as in other layered glasses but a correspondence with lower amounts of zinc has also been determined. Both elements appear enriched in the red coloured layers, while the other quantified elements have the same relative composition along the sample. Corrosion layers, due to the lead supporting structure of the window, were also found.

  1. Modified Alizarin Red S-Alcian Blue Staining for Reptilian Skeleton

    Directory of Open Access Journals (Sweden)

    Muhammad Ja’far Luthfi

    2016-04-01

    Full Text Available Skeletal staining is an important method in anatomical study. The aim of the research was to develop staining and clearing method of Reptilian skeleton using Alizarin Red S-Alcian Blue. The specimen were eviscerated, fixed, stained, cleared, and keep in glycerine solution. This modified double-staining has successfully stain bone and cartilage of Reptilian.

  2. The Effects of Trifluralin on LH, FSH and Testosterone Hormone Levels and Testis Histological Changes in Adult Rats

    Directory of Open Access Journals (Sweden)

    Mehrdad Shariati

    2008-01-01

    Full Text Available Background: Trifluralin is a herbicide and used in agriculture widely. It enters plants throughdeveloping roots and stops plant cells from division and elongation (meristemic inhibitor.Extensive application of trifluralin to control annual grasses and broadleaf weeds in agriculture,horticulture and horn garden, leads to environmental pollution and its entrance into the food chaincould have determined effects on human and other species. In this research the effects of trifluralinon reproductive parameters of the male rats including serum luteinizing hormone (LH, folliclestimulatinghormone (FSH, testosterone levels, and changes in testicular tissue and body weightwere investigated.Materials and Methods: For this purpose male rats were randomly divided in 5 groups, to includecontrol, sham (received normal saline as a solvent, and three experimental groups which received500, 1000 and 2000 mg/kg oral trifluralin respectively. After 16 days, body and testis weight weremeasured and blood samples were taken from heart and used for measurement of LH, FSH andtestosterone levels. To evaluate histological changes, testes were removed and weighed and, afterobtaining tissue section, stained by HE.Results: Serum testosterone, FSH, and LH levels showed significant decrease in experimentalgroups (p<0.05. There was significant decrease in the number of germinal and somatic cells intestis in experimental groups. There was also a significant decrease in body and testis weight inexperimental groups.Conclusion: It can be concluded that oral administration of trifluralin could decrease gonadotropinsand testosterone hormone levels and also this herbicide could have hazardous effects on testistissue.

  3. Histological Study of the Effects of Ibuprofen on Acute Apical Inflammation Following Root Canal Therapy in Cats

    Directory of Open Access Journals (Sweden)

    Ghoddusi J

    2000-05-01

    Full Text Available Postoperative pain following Root Canal Therapy (RCT is the common problem for patients,"ndentist and dental staff. Regardless of cause it appears that postoperative pain and discomfort is related to"nperiapical inflammation. The purpose of this study was to investigate histologically the effect of"nIbuprofen on acute apical inflammation following RCT. on cat. Fifty-six cuspids teeth of 14 cats were"ndivided into three groups, 6 cats for experimental group, 6 cats for positive control group and 2 cats for"nnegative control group. In first group, to anesthesize the animals each cat received 10 mg/kg Ibuprofen"n36 hours before RCT and repeated every 24 hours. In second group RCT was performed in 24 cat teeth"nwithout any prescription. In negative control groups 2 cats, no RCT were done."nFollowing vital perfusion, in interval of 6, 12, 24 hours, samples were post fixed in 10% formalin and"nfive micron sections were prepared, and stained with H&E and studied under the Mann Whitney"nparameter test. Infilteration of inflamatory cells in 6 hours interval in experimental group was"nsignificantly less than two control groups. In 12 hours samples, however, inflammation in experimental"ngroup reduced, but it was not significant. In 24 hours samples there was no significant difference"nbetween experimental groups and controls. The result of this in-vivo study shows prescription of"nIbuprofen before root canal therapy can reduce inflammatory reaction.

  4. Histological studies on the regeneration of small-intestine epithelium of rats irradiated with sublethal doses of x rays

    Energy Technology Data Exchange (ETDEWEB)

    Lewicki, Z; Figurski, R; Sulikowska, A

    1975-01-01

    The dynamics of regeneration of small-intestine epithelium was studied in rats irradiated with x rays in sublethal doses of 550, 600, or 750 R. Sixty-two irradiated and 22 control animals were used in the experiment. They were killed 1, 2, 4, 6, 8, 14, and 25 days after the irradiation. Specimens of duodenum and jejunum were examined histologically, the sections being stained with H. E. and p.a.S. Already 1 and 2 days after irradiation the intestinal villi became shorter and deformed. The blood vessels were damaged, the enterocytes showed features of degeneration and vacuolization, the epithelium was detached by the exudate which accumulated in the strong. Irradiation markedly disturbed the regeneration of intestinal epithelium in the period from the 1st to the 6th day. Cytological calculations indicate tha on the 1st and 2nd days after irradiation the number of epithelial cells of the villi, and particularly of young cryptal ones, markedly dropped. On the 4th and 6th days increased proliferation of young cryptal cells considerably surpassed the physiological rate. The accompanying disturbances in differentiation consisted in a decreased acidophilic to basophilic cells ration and in retardation of maturation of goblet cells. The absolute number of goblet cells was increased, as well as their proportion to the number of enterocytes.

  5. Histologic characterization of canine dilated cardiomyopathy.

    Science.gov (United States)

    Tidholm, A; Jönsson, L

    2005-01-01

    Dilated cardiomyopathy (DCM), characterized by chamber dilatation and myocardial systolic and diastolic dysfunction, is one of the most common heart diseases in dogs. The clinical diagnosis is based on findings on echocardiographic and Doppler examinations, with the active exclusion of other acquired or congenital heart diseases. However, the echocardiographic criteria for the diagnosis of DCM are not wholly specific for the disease, and histologic examination may be necessary for final diagnosis. Review of reports on histologic findings in dogs with clinically diagnosed DCM reveals two histologically distinct forms of DCM: 1) cardiomyopathy of Boxers and Doberman Pinschers, corresponding to the "fatty infiltration-degenerative" type and 2) the form seen in many giant, large-, and medium-sized breeds, including some Boxers and Doberman Pinschers, classified as the "attenuated wavy fiber" type of DCM. The histologic changes of the attenuated wavy fiber type of DCM may precede clinical and echocardiographic signs of heart disease, thus indicating an early stage of DCM.

  6. SE Marine Mammal Histology/Tissue data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Tissue samples are collected from stranded marine mammals in the Southeastern United States. These tissue samples are examined histologically and evaluated to...

  7. Distortion of the temporary cavity and its influence on staining in firearm barrels.

    Science.gov (United States)

    Schyma, Christian; Müller, Rolf; Brenčičová, Eva; Brünig, Julia

    2018-06-01

    After contact shots to the head, biological traces can be found inside the barrel of the firearm. Experimental protocols to generate this sort of staining, using 12 cm gelatin cubes containing thin foil bags filled with acrylic paint, human blood, and radiocontrast agent, have been developed. Previous research on shots fired at a distance has shown the underlay sustaining these gelatin cubes has an influence on experimental results. This study was conducted to investigate the role of the sustaining base of the gelatin blocks during contact shots, and its influence on the staining result inside firearm barrels. Eighteen contact shots were performed using 22 LR, 32 ACP (7.65 Browning) and 9 mm Luger semi-automatic pistols. With each pistol, shots were fired onto six gelatin cubes; three placed upon a rigid platform and three upon an elastic underlay. The shots were recorded by a high-speed video camera as they penetrated the gelatin cube. Any staining present inside the firearm barrels after the shots were fired was documented by endoscopy. Cross sections of the gelatin blocks were then compared to the high-speed video. It was found that the nature of the staining inside the barrel was not influenced by the underlay sustaining the target model. In the experiment using a 9 mm Luger, the rigid counterfort provoked a visible distortion of the temporary cavity, but, cross sectional analysis of the gelatin cubes did not reveal a relevant influence of the sustaining underlay on the crack length in the gelatin. This could be explained by a secondary expansion of the temporary cavity left by the projectile as a consequence of subsequent inflow of muzzle gases.

  8. Histologic progression in non-hodgkin's lymphoma

    International Nuclear Information System (INIS)

    Hubbard, S.M.; Chabner, B.A.; DeVita, V.T. Jr.; Simon, R.; Berard, C.W.; Jones, R.B.; Garvin, A.J.; Canellos, G.P.; Osborne, C.K.; Young, R.C.

    1982-01-01

    The clinical course and biopsy specimens from 515 consecutive non-Hodgkin's lymphoma patients was evaluated retrospectively in an attempt to determine the clinical importance of documented changes in histology over time. Two-hundred and five of these patients has an initial diagnosis of nodular lymphoma and were reviewed for this anaysis. Sixty-three underwent a repeat biopsy greater than 6 mo after initial diagnosis. In 23 patients, these repeat biopsies revealed a change in histology to a diffuse pattern and/or a change to a larger ''histiocytic'' cell type, while repeat biopsies for the other 40 (63%) disclosd persistence of a nodular pattern and no clear change in basic cell type. Progression from nodular lymphoma to diffuse histiocytic, mixed, or undifferentiated types of lymphomas of Rappaport was found in repeate biopsies obtained from 19 patients (30%). Prognosis for survival following a biopsy that demonstrated histologic change was related to the histology demonstrated at the most recent biopsy and to the response to subsequent drug treatment. Survival following repeat biopsy for these 19 patients was significantly shorter than for the 40 patients whose histology remained nodular (p < 0.001). However, attainment of a complete remission with intensive combination chemotherapy was associated with prolonged survival in eight patients and prolonged disease-free survival in one patient. Since prior treatment may compromise the ability to achieve a complete response to chemotherapy in patients with nodular lymphoma who develop an aggressive diffuse histology, the likelihood of histologic progression must be considered in the design of future clinical trials in nodular lymphoma. Histologic progression does not preclude attainment of a complete response to intensive chemotherapy

  9. [Intrapartum amnioinfusion in patients with meconium-stained amniotic fluid].

    Science.gov (United States)

    Engel, Karina; Samborska, Monika; Bilar, Marek; Sipak-Szmigiel, Olimpia; Ronin-Walknowska, Elzbieta

    2008-09-01

    The aim of the study was to evaluate the effect of intrapartum amnioinfusion in the presence of meconium stained amniotic fluid. 93 women with meconium-stained amniotic fluid were assigned to receive amnioinfusion or no amnioinfusion (128 women). The trials were evaluated for fetal distress syndrome, route of delivery, fetal acidemia, Apgar score at 1 and 5 min., meconium aspiration syndrome, postpartum endometritis and maternal hospital stays. Amnioinfusion in cases of meconium-stained fluid did not improve the number of fetal distress symptoms during fetal heart rate monitoring. Amnioinfusion was associated with a significant decrease of neonatal acidemia although it did not improve Apgar score. In our study amnioinfusion was not associated with reduction in the incidence of neonatal outcome and puerperial complications.

  10. Indocyanine green staining facilitates detection of bleb leakage during trabeculectomy.

    Science.gov (United States)

    Okazaki, Teruhiko; Kiuchi, Takahiro; Kawana, Keisuke; Oshika, Tetsuro

    2007-03-01

    To report a new technique to visualize bleb leakage using indocyanine green (ICG) staining during trabeculectomy. The ICG solution was widely applied over the filtering bleb including the conjunctival wound before completion of trabeculectomy. This procedure was performed in 48 eyes of 44 consecutive patients undergoing trabeculectomy between December 2004 and October 2005. Without staining, bleb leakage was not identified by the direct observation under the operating microscope. ICG staining clearly visualized aqueous leakage from the bleb in 5 eyes (10.4%). The bleb leakage in these eyes was easily repaired with 10-0 nylon sutures, and no eyes, including these 5 cases, showed bleb leakage after surgery. There were no intraoperative and postoperative complications related to ICG application. The application of ICG during trabeculectomy is a simple and useful technique to facilitate detection and repair of the bleb leakage.

  11. MEGARA Optics: stain removal in PBM2Y prisms

    International Nuclear Information System (INIS)

    Aguirre-Aguirre, D; Izazaga-Pérez, R; Carrasco, E; Villalobos-Mendoza, B; De Paz, A Gil; Gallego, J; Iglesias, J

    2017-01-01

    MEGARA is the new integral-field and multi-object optical spectrograph for the GTC. For medium and high resolution, the dispersive elements are volume phase holographic gratings, sandwiched between two flat windows and two prisms of high optical precision. The prisms are made of Ohara PBM2Y optical glass. After the prisms polishing process, some stains appeared on the surfaces. For this, in this work is shown the comparative study of five different products (muriatic acid, paint remover, sodium hydroxide, aqua regia and rare earth liquid polish) used for trying to eliminate the stains of the HR MEGARA prisms. It was found that by polishing with the hands the affected area, and using a towel like a kind of pad, and polish during five minutes using rare earth, the stains disappear completely affecting only a 5% the rms of the surface quality. Not so the use of the other products that did not show any apparent result. (paper)

  12. Chromosome-specific staining to detect genetic rearrangements

    Energy Technology Data Exchange (ETDEWEB)

    Gray, Joe W.; Pinkel, Daniel; Tkachuk, Douglas; Westbrook, Carol

    2013-04-09

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  13. Improvement of malaria diagnostic system based on acridine orange staining.

    Science.gov (United States)

    Kimura, Masatsugu; Teramoto, Isao; Chan, Chim W; Idris, Zulkarnain Md; Kongere, James; Kagaya, Wataru; Kawamoto, Fumihiko; Asada, Ryoko; Isozumi, Rie; Kaneko, Akira

    2018-02-07

    Rapid diagnosis of malaria using acridine orange (AO) staining and a light microscope with a halogen lamp and interference filter was deployed in some malaria-endemic countries. However, it has not been widely adopted because: (1) the lamp was weak as an excitation light and the set-up did not work well under unstable power supply; and, (2) the staining of samples was frequently inconsistent. The halogen lamp was replaced by a low-cost, blue light-emitting diode (LED) lamp. Using a reformulated AO solution, the staining protocol was revised to make use of a concentration gradient instead of uniform staining. To evaluate this new AO diagnostic system, a pilot field study was conducted in the Lake Victoria basin in Kenya. Without staining failure, malaria infection status of about 100 samples was determined on-site per one microscopist per day, using the improved AO diagnostic system. The improved AO diagnosis had both higher overall sensitivity (46.1 vs 38.9%: p = 0.08) and specificity (99.0 vs 96.3%) than the Giemsa method (N = 1018), using PCR diagnosis as the standard. Consistent AO staining of thin blood films and rapid evaluation of malaria parasitaemia with the revised protocol produced superior results relative to the Giemsa method. This AO diagnostic system can be set up easily at low cost using an ordinary light microscope. It may supplement rapid diagnostic tests currently used in clinical settings in malaria-endemic countries, and may be considered as an inexpensive tool for case surveillance in malaria-eliminating countries.

  14. p16/ki-67 dual-stain cytology in the triage of ASCUS and LSIL papanicolaou cytology: results from the European equivocal or mildly abnormal Papanicolaou cytology study.

    Science.gov (United States)

    Schmidt, Dietmar; Bergeron, Christine; Denton, Karin J; Ridder, Ruediger

    2011-06-25

    The objective of this study was to analyze the diagnostic performance of a newly established immunocytochemical dual-stain protocol, which simultaneously detects p16(INK4a) and Ki-67 expression in cervical cytology samples, for identifying high-grade cervical intraepithelial neoplasia (CIN2+) in women with Papanicolaou (Pap) cytology results categorized as atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesions (LSIL). Residual liquid-based cytology material from 776 retrospectively collected ASCUS/LSIL cases that were available from a recent study evaluating p16 cytology and HPV testing were subjected to p16/Ki-67 dual staining. The presence of 1 or more double-immunoreactive cell(s) was regarded as a positive test outcome, irrespective of morphology. Test results were correlated to histology follow-up. Sensitivity of p16/Ki-67 dual-stain cytology for biopsy-confirmed CIN2+ was 92.2% (ASCUS) and 94.2% (LSIL), while specificity rates were 80.6% (ASCUS) and 68.0% (LSIL), respectively. Similar sensitivity/specificity profiles were found for both age groups of women aged aged ≥30 years. Dual-stain cytology showed comparable sensitivity, but significantly higher specificity, when compared with human papillomavirus (HPV) testing. The results of this study show that p16/Ki-67 dual-stain cytology provided a high sensitivity for the detection of underlying CIN2+ in women with ASCUS or LSIL Pap cytology results, comparable to the rates previously reported for HPV testing and p16 single-stain cytology. However, the specificity of this morphology-independent interpretation of p16/Ki-67 dual-stain cytology testing was further improved compared with the earlier p16 single-stain cytology approach, which required morphology interpretation, and it is significantly higher when compared with HPV testing. Copyright © 2011 American Cancer Society.

  15. TANTANGAN DAN PELUANG JURUSAN TADRIS DI IAIN/STAIN

    Directory of Open Access Journals (Sweden)

    Mohammad Kosim

    2009-01-01

    Full Text Available A consequence of dualism in education policy, the implementatin of tadris in IAIN often faces many problems. This paper will describe the history and the development of tadris in IAIN/STAIN. The discussion begins with the origin of tadris in IAIN, problems araise in the implementation of tadris in IAIN/STAIN, including the alumni’s problem. In addition, this article also reveals several opportunities for tadris graduation to develop madrasah as purely religious school to public islamic school as the result of the paradigm change.

  16. News from the Biological Stain Commission no. 12

    DEFF Research Database (Denmark)

    Lyon, H O

    2012-01-01

    In this 12(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the meetings of ISO/TC 212/WG 1 Quality and competence in the medical laboratory and ISO....../TC 212/WG 3 In vitro diagnostic products both held on 2 - 3 June 2010, plus information on the second plenary meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 4 June 2010. All meetings took place in Seoul, Republic of Korea. Finally, information is provided...

  17. Evaluation Method of Accumulated Cooking Oil Stains in Room

    OpenAIRE

    五十嵐, 由利子; 中村, 和吉; 萬羽, 郁子; Igarashi, Yuriko; Nakamura, Kazuyoshi; Banba, Ikuko

    2005-01-01

    The diffusion of the oil mist while cooking affects the entire room, leaving stains on the ceiling and walls. The validity of measuring the color difference of stains was examined by installing teflon plates in the kitchen and the adjacent space with a view to assessing the oil diffusion. Four houses were designated for the examination. In each house, four teflon plates (20×40mm) were installed on the ceiling and walls of the kitchen and the space adjacent to it. Then, one plate was removed a...

  18. MR imaging of multiple fibroadenoma in breast: comparison with color doppler images and histologic findings

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Soo Young; Yang, Ik; Park, Hai Jung; Lee, Yul; Chung, Bong Wha; Ahn, Hye Kyung [Hallym Univ. College of Medicine, Seoul (Korea, Republic of)

    1997-10-01

    To understand the different signal intensities seen on contrast enhanced magnetic resonance imaging (MRI) in multiple fibroadenoma of the breast, and to compare these with color Doppler ultrasonographic (CDUS) and histologic findings. MRI (1.0 Tesla, TIWI, T2WI, 3D-gradient echo dynamic contrast enhancement study) findings of 24 histologically proven cases of fibroadenoma in five patients were evaluated and compared with the histologic components (myxoid, adenomatous, fibrous). In addition, vascular flow, as seen on CDUS and histologic section, was compared. The observed degree of signal intensity waw classified into three groups, as follows: negative, 8.3%, mild to moderate, 54.2%; marked, 37.5%. On histologic section, the greater the fibrotic component, the higher the intensity of MRI enhancement, the greater the glandular component, and the intensity. CDUS showed vascular flow in only one tumor larger than 3cm in diameter. Vascular patterns of tumors on CDUS were dots in mass and detouring pattern, but in this case and in strongly enhanced cases, tumor vascularity-as seen on histologic section-showed no significant increase. Different signal intensities seen on contrast enhanced MRI in multiple fibroadenoma of the breast may be related more to the amount of glandular and fibrotic component than to increased tumor vascularity.

  19. MR imaging of multiple fibroadenoma in breast: comparison with color doppler images and histologic findings

    International Nuclear Information System (INIS)

    Chung, Soo Young; Yang, Ik; Park, Hai Jung; Lee, Yul; Chung, Bong Wha; Ahn, Hye Kyung

    1997-01-01

    To understand the different signal intensities seen on contrast enhanced magnetic resonance imaging (MRI) in multiple fibroadenoma of the breast, and to compare these with color Doppler ultrasonographic (CDUS) and histologic findings. MRI (1.0 Tesla, TIWI, T2WI, 3D-gradient echo dynamic contrast enhancement study) findings of 24 histologically proven cases of fibroadenoma in five patients were evaluated and compared with the histologic components (myxoid, adenomatous, fibrous). In addition, vascular flow, as seen on CDUS and histologic section, was compared. The observed degree of signal intensity waw classified into three groups, as follows: negative, 8.3%, mild to moderate, 54.2%; marked, 37.5%. On histologic section, the greater the fibrotic component, the higher the intensity of MRI enhancement, the greater the glandular component, and the intensity. CDUS showed vascular flow in only one tumor larger than 3cm in diameter. Vascular patterns of tumors on CDUS were dots in mass and detouring pattern, but in this case and in strongly enhanced cases, tumor vascularity-as seen on histologic section-showed no significant increase. Different signal intensities seen on contrast enhanced MRI in multiple fibroadenoma of the breast may be related more to the amount of glandular and fibrotic component than to increased tumor vascularity

  20. Immunohistochemical analysis of medullary breast carcinoma autoantigens in different histological types of breast carcinomas

    Directory of Open Access Journals (Sweden)

    Kostianets Olga

    2012-11-01

    Full Text Available Abstract Background On the past decade a plethora of investigations were directed on identification of molecules involved in breast tumorogenesis, which could represent a powerful tool for monitoring, diagnostics and treatment of this disease. In current study we analyzed six previously identified medullary breast carcinoma autoantigens including LGALS3BP, RAD50, FAM50A, RBPJ, PABPC4, LRRFIP1 with cancer restricted serological profile in different histological types of breast cancer. Methods Semi-quantitative immunohistochemical analysis of 20 tissue samples including medullary breast carcinoma, invasive ductal carcinoma, invasive lobular carcinoma and non-cancerous tissues obtained from patients with fibrocystic disease (each of five was performed using specifically generated polyclonal antibodies. Differences in expression patterns were evaluated considering percent of positively stained cells, insensitivity of staining and subcellular localization in cells of all tissue samples. Results All 6 antigens predominantly expressed in the most cells of all histological types of breast tumors and non-cancerous tissues with slight differences in intensity of staining and subcellular localization. The most significant differences in expression pattern were revealed for RAD50 and LGALS3BP in different histological types of breast cancer and for PABPC4 and FAM50A antigens in immune cells infiltrating breast tumors. Conclusions This pilot study made possible to select 4 antigens LGALS3BP, RAD50, PABPC4, and FAM50A as promising candidates for more comprehensive research as potential molecular markers for breast cancer diagnostics and therapy. Virtual slides The virtual slides’ for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1860649350796892

  1. Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria

    International Nuclear Information System (INIS)

    Fife, D.J.; Bruhn, D.F.; Miller, K.S.; Stoner, D.L.

    2000-01-01

    A fluorescence-labeled wheat germ agglutinin staining technique was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure

  2. An Original Approach for Quantification of Blood Vessels on the Whole Tumour Section

    Directory of Open Access Journals (Sweden)

    Nga Tran Kim

    2003-01-01

    Full Text Available Relative abundance of tumour angiogenesis has been shown to be of clinical relevance in cancers of various locations such as the ovary. Nevertheless, several problems are encountered when quantifying tumour microvessels: (i as many other tumour markers, vascularity pattern is often heterogeneous within the tumour mass and even within the same histological section. As a consequence, an adequate acquisition method must be developed for accurate field sampling. (ii Manual microvessel counting is long, tedious and subject to poor reproducibility. Introduction in routine practice requires a fast, reproducible and reliable automatic image processing. In this study we present an original procedure combining a slide scanner image acquisition and a fully automatic image analysis sequence. The slide scanner offers the advantage of recording an image of the whole histological section for subsequent automatic blood vessel detection and hot spot area location. Microvessel density and surface fraction were measured for the whole section as well as within hot spots. Different immunostaining methods were tested in order to optimise the procedure. Moreover, the method proposed was submitted to a quality control procedure, with reference to interactive identification of microvessels at scanner level. This experiment showed that 93 to 97% of blood vessels were detected, according to the staining protocol used. Colour figures can be viewed on http://www.esacp.org/acp/2003/25‐2/kim.htm.

  3. The human sunburn reaction: histologic and biochemical studies

    International Nuclear Information System (INIS)

    Gilchrest, B.A.; Soter, N.A.; Stoff, J.S.; Mihm, M.C. Jr.

    1981-01-01

    The ultraviolet-induced erythema reaction was investigated histologically and biochemically in four subjects, utilizing suction blister aspirates, analyzed for histamine and prostaglandin E2 (PGE2), and Epon-embedded 1-mu skin biopsy sections from control skin and from irradiated skin at intervals for 72 hours after exposure to a Hanovia lamp. Major histologic alterations in the epidermis included dyskeratotic and vacuolated keratinocytes (sunburn cells), and disappearance of Langerhans cells. In the dermis the major changes were vascular, involving both the superficial and deep venular plexuses. Endothelial cell enlargement was first apparent within 30 minutes of irradiation, peaked at 24 hours, and persisted throughout the 72-hour study period. Mast cell degranulation and associated perivenular edema were first apparent at 1 hour and striking at the onset of erythema, 3 to 4 hours postirradiation; edema was absent and mast cells were again normal in number and granule content at 24 hours. Histamine levels rose approximately fourfold above control values immediately after the onset of erythema and returned to baseline within 24 hours. PGE2 levels were statistically elevated even before the onset of erythema and reached approximately 150% of the control value at 24 hours. These data provide the first evidence that histamine may mediate the early phase of the human sunburn reaction and increase our understanding of its complex histologic and biochemical sequelae

  4. Electron Beam Induced Mass Loss Dependence on Stained Thin Epon Resin Sections

    Czech Academy of Sciences Publication Activity Database

    Skoupý, Radim; Nebesářová, Jana; Krzyžánek, Vladislav

    2016-01-01

    Roč. 22, S3 (2016), s. 926-927 ISSN 1431-9276 R&D Projects: GA ČR(CZ) GA14-20012S; GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 ; RVO:60077344 Keywords : TEM * STEM * EFTEM Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering; JA - Electronics ; Optoelectronics, Electrical Engineering (BC-A) Impact factor: 1.891, year: 2016

  5. Brazilwood, sappanwood, brazilin and the red dye brazilein: from textile dyeing and folk medicine to biological staining and musical instruments.

    Science.gov (United States)

    Dapson, R W; Bain, C L

    2015-01-01

    Brazilin is a nearly colorless dye precursor obtained from the heartwood of several species of trees including brazilwood from Brazil, sappanwood from Asia and the Pacific islands, and to a minor extent from two other species in Central America, northern South America and the Caribbean islands. Its use as a dyeing agent and medicinal in Asia was recorded in the 2(nd) century BC, but was little known in Europe until the 12(th) century AD. Asian supplies were replaced in the 16(th) century AD after the Portuguese discovered vast quantities of trees in what is now Brazil. Overexploitation decimated the brazilwood population to the extent that it never fully recovered. Extensive environmental efforts currently are underway to re-create a viable, sustainable population. Brazilin is structurally similar to the better known hematoxylin, thus is readily oxidized to a colored dye, brazilein, which behaves like hematein. Attachment of the dye to fabric is by hydrogen bonding or in conjunction with certain metallic mordants by coordinative bonding. For histology, most staining procedures involve aluminum (brazalum) for staining nuclei. In addition to textile dyeing and histological staining, brazilin and brazilein have been and still are used extensively in Asian folk medicine to treat a wide variety of disorders. Recent pharmacological studies for the most part have established a scientific basis for these uses and in many cases have elucidated the biochemical pathways involved. The principal use of brazilwood today is for the manufacture of bows for violins and other stringed musical instruments. The dye and other physical properties of the wood combine to produce bows of unsurpassed tonal quality.

  6. Pre-staining thin layer chromatography method for amino acid ...

    African Journals Online (AJOL)

    Jane

    2010-12-13

    Dec 13, 2010 ... inexpensive and the results obtained were clean and reproducible. However, it is suitable for the high throughput screening of amino acid-producing strains. Key words: Thin layer chromatography, pre-staining, amino acid detection. INTRODUCTION. Several analytical techniques have been often used for.

  7. Color and dichroism of silver-stained glasses

    International Nuclear Information System (INIS)

    Molina, Gloria; Murcia, Sonia; Molera, Judit; Roldan, Clodoaldo; Crespo, Daniel; Pradell, Trinitat

    2013-01-01

    Yellow decorations in glasses have been produced since the beginning of the fourteenth century by incorporating metallic silver nanoparticles into the glass (from a few to some tens of nanometers). The optical response of the glass-particles composite is determined by the surface plasmon resonance absorption and scattering of the nanometric metallic particles. Generally, the same color is perceived in reflection and in transmission although dichroic effects are occasionally observed. As silver-stained glasses were designed to be observed in transmission, tuning the transmission color from yellow to red was of technological interest. The relationship between the color observed both in transmission and reflection and the composition and nanostructure of regular (yellow) and dichroic (yellow and red) silver stains from the Renaissance (late fifteenth and sixteenth century, respectively) is related to the presence of a layer (of about 10–20 μm thick) of metallic silver nanoparticles (from few to 100 nm in size). The correlation between the colors observed and the silver stain nanostructure is studied with particular emphasis on the origin of the dichroic behavior. The optical response is computed and compared to the experimental data. Differences in the synthesis parameters responsible for the colors and for the dichroic behavior of the silver stain glasses are proposed. This is essential for the replication of the glass pieces which are required as replacements in the restoration/conservation of the windows but is also of broader interest

  8. Alternate gram staining technique using a fluorescent lectin.

    Science.gov (United States)

    Sizemore, R K; Caldwell, J J; Kendrick, A S

    1990-01-01

    Fluorescence-labeled wheat germ agglutinin binds specifically to N-acetylglucosamine in the outer peptidoglycan layer of gram-positive bacteria. The peptidoglycan layer of gram-negative bacteria is covered by a membrane and is not labeled by the lectin. By exploiting this phenomenon, an alternative Gram staining technique has been developed. Images PMID:1697149

  9. Black stain and dental caries in Filipino schoolchildren.

    NARCIS (Netherlands)

    Heinrich-Weltzien, R.; Monse, B.; Palenstein Helderman, W.H. van

    2009-01-01

    Black stain is defined as dark pigmented exogenous substance in lines or dots parallel to the gingival margin and firmly adherent to the enamel at the cervical third of the tooth crowns in the primary and permanent dentition. OBJECTIVES: This study was conducted to assess the prevalence of black

  10. Amalgam stained dentin: a proper substrate for bonding resin composite?

    NARCIS (Netherlands)

    Scholtanus, J.D.

    2016-01-01

    Nowadays the use of dental amalgam is mostly abandoned and substituted by tooth colored resin composites that can be bonded to teeth tissues by adhesive techniques. The aim of this thesis was to find out whether dark stained dentin, as often observed after removal of amalgam restorations and

  11. Color and dichroism of silver-stained glasses

    Energy Technology Data Exchange (ETDEWEB)

    Molina, Gloria [Universitat Politecnica de Catalunya, Center for Research in NanoEngineering (Spain); Murcia, Sonia [Universidad de Valencia, Instituto de Ciencia de los Materiales (Spain); Molera, Judit [Universitat de Vic, GRTD, Escola Politecnica Superior (Spain); Roldan, Clodoaldo [Universidad de Valencia, Instituto de Ciencia de los Materiales (Spain); Crespo, Daniel; Pradell, Trinitat, E-mail: Trinitat.Pradell@upc.edu [Universitat Politecnica de Catalunya, Center for Research in NanoEngineering (Spain)

    2013-09-15

    Yellow decorations in glasses have been produced since the beginning of the fourteenth century by incorporating metallic silver nanoparticles into the glass (from a few to some tens of nanometers). The optical response of the glass-particles composite is determined by the surface plasmon resonance absorption and scattering of the nanometric metallic particles. Generally, the same color is perceived in reflection and in transmission although dichroic effects are occasionally observed. As silver-stained glasses were designed to be observed in transmission, tuning the transmission color from yellow to red was of technological interest. The relationship between the color observed both in transmission and reflection and the composition and nanostructure of regular (yellow) and dichroic (yellow and red) silver stains from the Renaissance (late fifteenth and sixteenth century, respectively) is related to the presence of a layer (of about 10-20 {mu}m thick) of metallic silver nanoparticles (from few to 100 nm in size). The correlation between the colors observed and the silver stain nanostructure is studied with particular emphasis on the origin of the dichroic behavior. The optical response is computed and compared to the experimental data. Differences in the synthesis parameters responsible for the colors and for the dichroic behavior of the silver stain glasses are proposed. This is essential for the replication of the glass pieces which are required as replacements in the restoration/conservation of the windows but is also of broader interest.

  12. A comparison study of histochemical staining of various tissues after ...

    African Journals Online (AJOL)

    mean =5.28) then Carnoy's (mean = 4.00). For Alcian blue and Perl's Prussian blue, the best staining qualities were obtained by Formalin (mean = 4.76 and 5.64 respectively) followed by Carnoy's (mean = 2.88 and 3.92 respectively).

  13. Christendom's Narratives and the Stained Glass Designs of Yusuf ...

    African Journals Online (AJOL)

    This paper attempts a recast of Christendom's narratives in the stained glass designs of Yusuf Cameron Adebayo Grillo as the distinctive overarching mechanism of the evangelisation paradigm of the post Vatican II Church. It, therefore, draws attention to the delimitation of time frames in the history of the art form. Using the ...

  14. Comparison of various staining techniques in the diagnosis of ...

    African Journals Online (AJOL)

    Journal of Medical and Biomedical Sciences ... This can be achieved via various diagnostic techniques, commonly microscopy in this environment, hence the need to compare the efficacy of the commonly ... The objective of the study is to identify the most effective of the commonly used stains in identifying these parasites.

  15. Laser beam diameter for port wine stain treatment

    NARCIS (Netherlands)

    Keijzer, M.; Pickering, J. W.; van Gemert, M. J.

    1991-01-01

    Optimal port wine stain treatment requires the selective absorption of light by the ectatic blood vessels. We investigated whether deeper blood vessels can be coagulated, without damaging other cutaneous structures, by varying the laser beam diameter. The penetration of the light was simulated with

  16. Borax methylene blue: a spectroscopic and staining study.

    Science.gov (United States)

    Donaldson, P T; Russo, A; Reynolds, C; Lillie, R D

    1978-07-01

    Borax methylene blue is quite stable at room temperatures of 22-25 C. At 30 C polychroming is slow; during 50 days in a water bath at this temperature the absorption peak moves from 665 to 656 nm. At 35 C, the absorption peak reaches 660 nm in 7 days, 654 nm in 14. At 60 C polychroming is rapid, the absorption peak reaching 640-620 nm in 3 days. When the pH of the borax methylene blue solutions, normally about 9.0, is adjusted to pH 6.5, the absorption peak remains at 665 nm even when incubated at 60 C for extended periods. When used as a blood stain 0.4 ml borax methylene blue (1% methylene blue in 1% borax), 4 ml acetone, 2 ml borax-acid phosphate buffer to bring the solution to pH 6.5, and distilled water to make 40 ml, with 0.2 ml 1% eosin added just before using, an excellent Nocht-Giemsa type stain is achieved after 30 minutes staining. The material plasmodia P. falciparum, P. vivax, and P. berghei stain moderate blue with dark red chromatin and green to black pigment granules. The study confirms Malachowski's 1891 results and explains Gautier's 1896-98 failure to duplicate it.

  17. a comparison of modified and standard papanicolaou staining ...

    African Journals Online (AJOL)

    2011-07-07

    Jul 7, 2011 ... modified pap method and standard Papanicolaou method respectively. The staining characteristics in .... alcohol was replaced by 0.5 % acetic acid and also, .... was 37.1, standard deviation of 8.0 and a median of. 36.5 years.

  18. Interlaboratory variability of Ki67 staining in breast cancer

    NARCIS (Netherlands)

    Focke, Cornelia M.; Bürger, Horst; van Diest, Paul J.; Finsterbusch, Kai; Gläser, Doreen; Korsching, Eberhard; Decker, Thomas; Anders, M.; Bollmann, R.; Eiting, Fr; Friedrich, K.; Habeck, J. O.; Haroske, G.; Hinrichs, B.; Behrens, A.; Krause, Lars Udo; Braun-Lang, U.; Lorenzen, J.; Minew, N.; Mlynek-Kersjes, M.; Nenning, H.; Packeisen, J.; Poche-de Vos, F.; Reyher-Klein, S.; Rothacker, D.; Schultz, M.; Sturm, U.; Tawfik, M.; Berghäuser, K. H.; Böcker, W; Cserni, G.; Habedank, S.; Lax, S.; Moinfar, F.; Regitnig, P.; Reiner-Concin, A.; Rüschoff, J.; Varga, Z.; Woziwodski, J.

    2017-01-01

    Background Postanalytic issues of Ki67 assessment in breast cancers like counting method standardisation and interrater bias have been subject of various studies, but little is known about analytic variability of Ki67 staining between pathology labs. Our aim was to study interlaboratory variability

  19. Comparison of different stains in imprint cytology with the conventional diagnostic tools in detection of Helicobacter pylori infection

    Directory of Open Access Journals (Sweden)

    Shashikant Adlekha

    2015-01-01

    Full Text Available Background: Helicobacter pylori (H. pylori infection is one of the most common causes of gastrointestinal morbidity and mortality worldwide. Early diagnosis of infection is imperative for the cure of distressing symptoms and prevention of complications. Objective: The objective of the study is to evaluate imprint cytology of gastric biopsy specimens as a rapid and cost-effective diagnostic tool and its comparison with conventional diagnostic tests. Materials and Methods: Antral biopsies were collected from 130 patients and evaluated for H. pylori infection by imprint cytology and histopathological examination by different stains and rapid urease test (RUT. Histopathological features of gastric biopsy specimens were also assessed. Results: A total of 118 patients showed H. pylori infection by two or more methods. Giemsa histology showed highest sensitivity, specificity, positive predictive value, negative predictive value and Youden′s Index (YI. Among imprint cytology stain methods, toluidine blue showed highest sensitivity and highest YI was obtained for PAP stain. Conclusion: Helicobacter pylori infection is associated with gastric mucosa changes like chronic active gastritis, atrophy, intestinal metaplasia, ulceration and carcinoma. Imprint cytology has high sensitivity and comparable predictive values to conventional diagnostic tools-histopathological examination and RUT in the detection of H. pylori infection.

  20. Effect of amnioinfusion for meconium stained amniotic fluid on perinatal outcome.

    Science.gov (United States)

    Ashfaq, F; Shah, A A

    2004-06-01

    To see the effect of amnioinfusion on perinatal outcome in cases of meconium staining of liquor. This study was conducted in department of Obstetrics and Gynaecology, unit 1, Jinnah Postgraduate Medical Centre, Karachi, from 1st January 1998 to 31st December 2000. Four hundred patients were included in this study, assigning 200 for amnioinfusion and 200 as control. All patients were matched in both the groups with respect to age, antenatal booking, parity, gestational age, stage of labour, colour of amniotic fluid and fetal birth weight. Both the groups were found to be comparable. The rate of Caesarean section was found to be 37% in amnioinfusion group, which collaborates with other international studies. The fetal outcome was better i.e. 91% alive and healthy, after amnioinfusion due to dilution of meconium stained amniotic fluid with physiological solutions. The perinatal outcome was recorded by Apgar score at 5 minutes. The perinatal morbidity and mortality both were significantly lowered and was found to be 6% as compared to 14% in control, which was also noticed by less number of admissions in nursery i.e. 12% and perinatal deaths. The incidence of meconium aspiration syndrome was found to be 56% in control and was reduced to 22% after amnioinfusion in the other arm of the study. These results are very encouraging and suggestion can be safely made that in future amnioinfusion will be the ideal method of preventing fetal distress due to meconium stained amniotic fluid.

  1. Comparative digital cartilage histology for human and common osteoarthritis models

    Directory of Open Access Journals (Sweden)

    Pedersen DR

    2013-02-01

    Full Text Available Douglas R Pedersen, Jessica E Goetz, Gail L Kurriger, James A MartinDepartment of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA, USAPurpose: This study addresses the species-specific and site-specific details of weight-bearing articular cartilage zone depths and chondrocyte distributions among humans and common osteoarthritis (OA animal models using contemporary digital imaging tools. Histological analysis is the gold-standard research tool for evaluating cartilage health, OA severity, and treatment efficacy. Historically, evaluations were made by expert analysts. However, state-of-the-art tools have been developed that allow for digitization of entire histological sections for computer-aided analysis. Large volumes of common digital cartilage metrics directly complement elucidation of trends in OA inducement and concomitant potential treatments.Materials and methods: Sixteen fresh human knees, 26 adult New Zealand rabbit stifles, and 104 bovine lateral plateaus were measured for four cartilage zones and the cell densities within each zone. Each knee was divided into four weight-bearing sites: the medial and lateral plateaus and femoral condyles.Results: One-way analysis of variance followed by pairwise multiple comparisons (Holm–Sidak method at a significance of 0.05 clearly confirmed the variability between cartilage depths at each site, between sites in the same species, and between weight-bearing articular cartilage definitions in different species.Conclusion: The present study clearly demonstrates multisite, multispecies differences in normal weight-bearing articular cartilage, which can be objectively quantified by a common digital histology imaging technique. The clear site-specific differences in normal cartilage must be taken into consideration when characterizing the pathoetiology of OA models. Together, these provide a path to consistently analyze the volume and variety of histologic slides necessarily generated

  2. Echocardiographic versus histologic findings in Marfan syndrome.

    Science.gov (United States)

    Gu, Xiaoyan; He, Yihua; Li, Zhian; Han, Jiancheng; Chen, Jian; Nixon, J V Ian

    2015-02-01

    This retrospective study attempted to establish the prevalence of multiple-valve involvement in Marfan syndrome and to compare echocardiographic with histopathologic findings in Marfan patients undergoing valvular or aortic surgery. We reviewed echocardiograms of 73 Marfan patients who underwent cardiovascular surgery from January 2004 through October 2009. Tissue histology was available for comparison in 29 patients. Among the 73 patients, 66 underwent aortic valve replacement or the Bentall procedure. Histologic findings were available in 29 patients, all of whom had myxomatous degeneration. Of 63 patients with moderate or severe aortic regurgitation as determined by echocardiography, 4 had thickened aortic valves. The echocardiographic findings in 18 patients with mitral involvement included mitral prolapse in 15. Of 11 patients with moderate or severe mitral regurgitation as determined by echocardiography, 4 underwent mitral valve repair and 7 mitral valve replacement. Histologic findings among mitral valve replacement patients showed thickened valve tissue and myxomatous degeneration. Tricuspid involvement was seen echocardiographically in 8 patients, all of whom had tricuspid prolapse. Two patients had severe tricuspid regurgitation, and both underwent repair. Both mitral and tricuspid involvement were seen echocardiographically in 7 patients. Among the 73 patients undergoing cardiac surgery for Marfan syndrome, 66 had moderate or severe aortic regurgitation, although their valves manifested few histologic changes. Eighteen patients had mitral involvement (moderate or severe mitral regurgitation, prolapse, or both), and 8 had tricuspid involvement. Mitral valves were most frequently found to have histologic changes, but the tricuspid valve was invariably involved.

  3. Clinical and Histologic Mimickers of Celiac Disease.

    Science.gov (United States)

    Kamboj, Amrit K; Oxentenko, Amy S

    2017-08-17

    Celiac disease is an autoimmune disorder of the small bowel, classically associated with diarrhea, abdominal pain, and malabsorption. The diagnosis of celiac disease is made when there are compatible clinical features, supportive serologic markers, representative histology from the small bowel, and response to a gluten-free diet. Histologic findings associated with celiac disease include intraepithelial lymphocytosis, crypt hyperplasia, villous atrophy, and a chronic inflammatory cell infiltrate in the lamina propria. It is important to recognize and diagnose celiac disease, as strict adherence to a gluten-free diet can lead to resolution of clinical and histologic manifestations of the disease. However, many other entities can present with clinical and/or histologic features of celiac disease. In this review article, we highlight key clinical and histologic mimickers of celiac disease. The evaluation of a patient with serologically negative enteropathy necessitates a carefully elicited history and detailed review by a pathologist. Medications can mimic celiac disease and should be considered in all patients with a serologically negative enteropathy. Many mimickers of celiac disease have clues to the underlying diagnosis, and many have a targeted therapy. It is necessary to provide patients with a correct diagnosis rather than subject them to a lifetime of an unnecessary gluten-free diet.

  4. [Improvement of Phi bodies stain and its clinical significance].

    Science.gov (United States)

    Gong, Xu-Bo; Lu, Xing-Guo; Yan, Li-Juan; Xiao, Xi-Bin; Wu, Dong; Xu, Gen-Bo; Zhang, Xiao-Hong; Zhao, Xiao-Ying

    2009-02-01

    The aim of this study was to improve the dyeing method of hydroperoxidase (HPO), to analyze the morphologic features of Phi bodies and to evaluate the clinical application of this method. 128 bone marrow or peripheral blood smears from patients with myeloid and lymphoid malignancies were stained by improved HPO staining. The Phi bodies were observed with detection rate of Phi bodies in different leukemias. 69 acute myeloid leukemia (AML) specimens were chosen randomly, the positive rate and the number of Phi bodies between the improved HPO and POX stain based on the same substrate of 3, 3'diaminobenzidine were compared. The results showed that the shape of bundle-like Phi bodies was variable, long or short. while the nubbly Phi bodies often presented oval and smooth. Club-like Phi bodies were found in M(3). The detection rates of bundle-like Phi bodies in AML M(1)-M(5) were 42.9% (6/14), 83.3% (15/18), 92.0% (23/25), 52.3% (11/21), 33.3% (5/15) respectively, and those of nubbly Phi bodies were 28.6% (4/14), 66.7% (12/18), 11.1% (3/25), 33.3% (7/21), 20.0% (3/15) respectively. The detection rate of bundle-like Phi bodies in M(3) was significantly higher than that in (M(1) + M(2)) or (M(4) + M(5)) groups. The detection rate of nubbly Phi bodies in (M(1) + M(2)) group was higher than that in M(3) group. In conclusion, after improvement of staining method, the HPO stain becomes simple, the detection rate of Phi bodies is higher than that by the previous method, the positive granules are more obvious, and the results become stable. This improved method plays an important role in differentiating AML from ALL, subtyping AML, and evaluating the therapeutic results.

  5. Histological organization of the female queen Devario regina (Fowler, 1934 during its juvenile stage

    Directory of Open Access Journals (Sweden)

    Piyakorn Boonyoung

    2016-02-01

    Full Text Available Limited research has been reported in the basic information about the structural organizations of fish organs in their juvenile state that could be used as histopathological biomarkers. Thus, the histological structures of important organs of the female fish Devario regina (Fowler, 1934 during its juvenile stage were exclusively examined using histological and histochemical approaches. Specimens were collected during the fishing season (July and October 2010 from the Tapee river, Thailand. Using histological analysis, the digestive system was distinctly composed of two parts; the digestive tract and accessory organs (liver and pancreas. Based on their histological structure, the epithelial organization of the oral cavity and pharynx was lined by stratified epithelium whereas the intestine was covered by a simple columnar epithelium and contained several goblet cells. The goblet cells were negatively stained with hematoxylin and eosin (H&E and Masson’s Trichrome (MT. In contrast, they were positively stained with Periodic Acid Schiff reaction (PAS and aniline blue (AB. The liver tissue in this fish was composed of polyhedral hepatocytes, with their sinusoids being distinctly located between the hepatocytes. The sinusoids were lined by a simple squamous epithelium. The pancreatic parenchyma mainly consisted of pyramidal cells that rested on a basal lamina in the acinar. Moreover, the pancreatic cells had a basophilic cytoplasm, a distinct basal nucleus and contained large eosinophilic zymogen granules. The excretory system especially referred to the kidney, and was composed of renal tubules and hematopoietic tissue. The female reproductive system was the ovary that was surrounded by a tunica albuginea. The ovary contained oocytes at differential stages of development including oogonia and the previtellogenic stage. Finally, the integument of this species consisted mainly of three layers of epidermis, dermis and hypodermis, respectively.

  6. Detection, mapping, and quantification of single walled carbon nanotubes in histological specimens with photoacoustic microscopy.

    Science.gov (United States)

    Avti, Pramod K; Hu, Song; Favazza, Christopher; Mikos, Antonios G; Jansen, John A; Shroyer, Kenneth R; Wang, Lihong V; Sitharaman, Balaji

    2012-01-01

    In the present study, the efficacy of multi-scale photoacoustic microscopy (PAM) was investigated to detect, map, and quantify trace amounts [nanograms (ng) to micrograms (µg)] of SWCNTs in a variety of histological tissue specimens consisting of cancer and benign tissue biopsies (histological specimens from implanted tissue engineering scaffolds). Optical-resolution (OR) and acoustic-resolution (AR)--Photoacoustic microscopy (PAM) was employed to detect, map and quantify the SWCNTs in a variety of tissue histological specimens and compared with other optical techniques (bright-field optical microscopy, Raman microscopy, near infrared (NIR) fluorescence microscopy). Both optical-resolution and acoustic-resolution PAM, allow the detection and quantification of SWCNTs in histological specimens with scalable spatial resolution and depth penetration. The noise-equivalent detection sensitivity to SWCNTs in the specimens was calculated to be as low as ∼7 pg. Image processing analysis further allowed the mapping, distribution, and quantification of the SWCNTs in the histological sections. The results demonstrate the potential of PAM as a promising imaging technique to detect, map, and quantify SWCNTs in histological specimens, and could complement the capabilities of current optical and electron microscopy techniques in the analysis of histological specimens containing SWCNTs.

  7. Detection, mapping, and quantification of single walled carbon nanotubes in histological specimens with photoacoustic microscopy.

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    Pramod K Avti

    Full Text Available In the present study, the efficacy of multi-scale photoacoustic microscopy (PAM was investigated to detect, map, and quantify trace amounts [nanograms (ng to micrograms (µg] of SWCNTs in a variety of histological tissue specimens consisting of cancer and benign tissue biopsies (histological specimens from implanted tissue engineering scaffolds.Optical-resolution (OR and acoustic-resolution (AR--Photoacoustic microscopy (PAM was employed to detect, map and quantify the SWCNTs in a variety of tissue histological specimens and compared with other optical techniques (bright-field optical microscopy, Raman microscopy, near infrared (NIR fluorescence microscopy.Both optical-resolution and acoustic-resolution PAM, allow the detection and quantification of SWCNTs in histological specimens with scalable spatial resolution and depth penetration. The noise-equivalent detection sensitivity to SWCNTs in the specimens was calculated to be as low as ∼7 pg. Image processing analysis further allowed the mapping, distribution, and quantification of the SWCNTs in the histological sections.The results demonstrate the potential of PAM as a promising imaging technique to detect, map, and quantify SWCNTs in histological specimens, and could complement the capabilities of current optical and electron microscopy techniques in the analysis of histological specimens containing SWCNTs.

  8. Randomised trial of amnioinfusion during labour with meconium stained amniotic fluid.

    Science.gov (United States)

    Rathor, Asmita Muthal; Singh, Ruchira; Ramji, S; Tripathi, Reva

    2002-01-01

    To assess the effect of amnioinfusion during labour with meconium stained amniotic fluid on caesarean section rate and perinatal outcome. Prospective randomised controlled study. A tertiary care teaching hospital in India. Women in labour at term with meconium stained amniotic fluid. Two hundred women in labour with > or = 37 weeks gestation, single cephalic presentation with moderate or thick meconium were randomised to control and amnioinfusion groups at a 1:1 ratio. Amnioinfusion was performed using 500 mL of normal saline over a period of 30 minutes in a study group. The control group received routine care. Both groups had intermittent auscultation of fetal heart rate during labour. The primary outcome measure was caesarean section rate. Secondary outcome measures were meconium aspiration syndrome, 1 minute and 5 minute apgar amnioinfusion group was less than the control group (RR 0.47; 95% CI 0.24-0.93). Amnioinfusion was associated with a significant decrease in the incidence of meconium at the vocal cords (P = 0.001); improvement in 1 minute apgar scores (P Amnioinfusion in an under resourced labour ward decreases caesarean section rates and fetal morbidity.

  9. Immunocytochemical detection of astrocytes in brain slices in combination with Nissl staining.

    Science.gov (United States)

    Korzhevskii, D E; Otellin, V A

    2005-07-01

    The present study was performed to develop a simple and reliable method for the combined staining of specimens to allow the advantages of immunocytochemical detection of astrocytes and assessment of the functional state of neurons by the Nissl method to be assessed simultaneously. The protocol suggested for processing paraffin sections allows preservation of tissue structure at high quality and allows the selective identification of astrocytes with counterstaining of neurons by the Nissl method. The protocol can be used without modification for processing brain specimens from humans and various mammals--except mice and rabbits.

  10. Rat intermediate lobe in culture: a histological and biochemical characterization.

    Science.gov (United States)

    Chronwall, B M; Bishop, J F; Gehlert, D R

    1988-01-01

    The histology, immunohistochemistry, peptide synthesis and secretion as well as proliferation rate of rat intermediate lobe (IL) were studied in primary cultures. The cultures contained two populations of cells: melanotrophs either organized in free floating lobules or in lobules which attached to the dishes and formed a monolayer. Both populations retained their in vivo morphology: polyhedral cells with smooth, ovoid nuclei and a large number of cytoplasmic secretory coated vesicles, a well developed Golgi apparatus, abundant mitochondria and extensive areas of rough endoplasmic reticulum. The melanotrophs stained with varying intensity for alpha-MSH and in situ hybridization showed the presence of pro-opiomelanocortin (POMC) mRNA. 35S-methionine incorporation combined with 2-D gel electrophoresis demonstrated POMC peptide synthesis and radioimmunoassay confirmed its secretion into the medium. 3H-thymidine uptake in the attached melanotrophs was considerably higher than that in the free-floating melanotrophs, demonstrating the dependency of proliferation rate on the cytoarchitecture of the explant. The retention of melanotroph morphology, biosynthetic and proliferative capacity in vitro affords a valid model system for studying POMC gene expression.

  11. Radioresponse of thymomas verified with histologic reponse

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    Ohara, Kiyoshi; Tatsuzaki, Hideo; Okumura, Toshiyuki; Itai, Yuji [Dept. of Radiology, Tsukuba Univ., Tsukuba City (Japan)]|[Inst. of Clinical Medicine, Tsukuba Univ., Tsukuba City (Japan); Fuji, Hiroshi [Dept. of Radiology, Tsukuba Univ., Tsukuba City (Japan); Sugahara, Shinji [Dept. of Radiology, Hitachi General Hospital, Hitachi City (Japan); Akaogi, Eiichi; Onizuka, Masataka; Ishikawa, Shigemi; Mitsui, Kiyofumi [Dept. of Surgery, Tsukuba Univ., Tsukuba City (Japan)]|[Inst. of Clinical Medicine, Tsukuba Univ., Tsukuba City (Japan)

    1998-12-31

    Patterns of radiologic response of 10 thymomas treated by preoperative radiotherapy (RT) (18-20 Gy/2 weeks) were determined in conjunction with histologic response. Changes in tumor volume were evaluated with CT scans obtained 5 to 36 days before and 14 to 24 days after the initiation of RT and before surgery. The extent of tumor volume reduction (TR) varied widely (40-78%), while the mean daily volume decrement expressed as a percentage of the pre-RT tumor volume correlated significantly with the pre-RT tumor volume. Histologically, the tumors, all of which were resected 17 to 33 days after RT initiation, generally consisted of predominant fibrous tissues, rare necrotic foci, and few epithelial cells. The TR did not correlate with pre-RT tumor volume, observation period, histologic subtype, or quantity of remaining epithelial cells. The TR of thymomas does not predict RT impact on tumor cells but does reflect the quantity of inherent tumor stroma. (orig.)