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Sample records for stained histological sections

  1. Histological stain evaluation for machine learning applications

    Directory of Open Access Journals (Sweden)

    Jimmy C Azar

    2013-01-01

    Full Text Available Aims: A methodology for quantitative comparison of histological stains based on their classification and clustering performance, which may facilitate the choice of histological stains for automatic pattern and image analysis. Background: Machine learning and image analysis are becoming increasingly important in pathology applications for automatic analysis of histological tissue samples. Pathologists rely on multiple, contrasting stains to analyze tissue samples, but histological stains are developed for visual analysis and are not always ideal for automatic analysis. Materials and Methods: Thirteen different histological stains were used to stain adjacent prostate tissue sections from radical prostatectomies. We evaluate the stains for both supervised and unsupervised classification of stain/tissue combinations. For supervised classification we measure the error rate of nonlinear support vector machines, and for unsupervised classification we use the Rand index and the F-measure to assess the clustering results of a Gaussian mixture model based on expectation-maximization. Finally, we investigate class separability measures based on scatter criteria. Results: A methodology for quantitative evaluation of histological stains in terms of their classification and clustering efficacy that aims at improving segmentation and color decomposition. We demonstrate that for a specific tissue type, certain stains perform consistently better than others according to objective error criteria. Conclusions: The choice of histological stain for automatic analysis must be based on its classification and clustering performance, which are indicators of the performance of automatic segmentation of tissue into morphological components, which in turn may be the basis for diagnosis.

  2. 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

  3. Fully automated registration of vibrational microspectroscopic images in histologically stained tissue sections.

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    Yang, Chen; Niedieker, Daniel; Grosserüschkamp, Frederik; Horn, Melanie; Tannapfel, Andrea; Kallenbach-Thieltges, Angela; Gerwert, Klaus; Mosig, Axel

    2015-11-25

    In recent years, hyperspectral microscopy techniques such as infrared or Raman microscopy have been applied successfully for diagnostic purposes. In many of the corresponding studies, it is common practice to measure one and the same sample under different types of microscopes. Any joint analysis of the two image modalities requires to overlay the images, so that identical positions in the sample are located at the same coordinate in both images. This step, commonly referred to as image registration, has typically been performed manually in the lack of established automated computational registration tools. We propose a corresponding registration algorithm that addresses this registration problem, and demonstrate the robustness of our approach in different constellations of microscopes. First, we deal with subregion registration of Fourier Transform Infrared (FTIR) microscopic images in whole-slide histopathological staining images. Second, we register FTIR imaged cores of tissue microarrays in their histopathologically stained counterparts, and finally perform registration of Coherent anti-Stokes Raman spectroscopic (CARS) images within histopathological staining images. Our validation involves a large variety of samples obtained from colon, bladder, and lung tissue on three different types of microscopes, and demonstrates that our proposed method works fully automated and highly robust in different constellations of microscopes involving diverse types of tissue samples.

  4. Lawsonia inermis And Hibiscus sabdariffa : Posible Histological Stains

    African Journals Online (AJOL)

    The ability of various concentrations of aqueous extracts of Lawsonia inermis and Hibiscus sabdariffa to stain histological tissues was demonstrated. The results with sections of tongue and kidney of the laboratory rat, cut at 6microns thickness showed that only the cellular cytoplasm was stained. However, combinations of ...

  5. A novel histological examination with dynamic three-dimensional reconstruction from multiple immunohistochemically stained sections of a PD-L1-positive colon cancer.

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    Korehisa, Shotaro; Ikeda, Tetsuo; Okano, Shinji; Saeki, Hiroshi; Oki, Eiji; Oda, Yoshinao; Hashizume, Makoto; Maehara, Yoshihiko

    2018-03-01

    Programmed cell death-ligand 1 (PD-L1) expression is observed in patients with microsatellite instability-high (MSI-H) colon cancer, which is susceptible to immune checkpoint blockade. The aim of this study was to investigate the interrelationship between PD-L1-positive cells and cytotoxic T cells, lymphatic vessels and vascular endothelium by using histological examination with the three-dimensional (3D) reconstruction of a PD-L1-positive colon cancer. Serial sections of MSI-H colon cancer tissue were stained with haematoxylin and eosin (H&E) and Masson trichrome stains; immunohistochemical analysis of PD-L1, CD8, D2-40 and CD31 was performed. Several 3D models of MSI-H colon cancer were reconstructed with a 3D data visualisation system. Moreover, 18 serial sections were stained with PD-L1, cytokeratin AE1/AE3, CD45, CD31, CD68 and H&E in the same case to confirm that PD-L1 was expressed on tumour cells, CD31-positive cells and macrophages in the invasive frontal region. Notably, there was a peak in the expression of PD-L1 and CD31 in the invasive frontal region. D2-40-positive cells were abundant in the overall tumour stroma, and CD8-positive cells infiltrated the tumour parenchyma. PD-L1 was expressed on tumour cells in the parenchyma and other cells in the stroma. Additional staining of 18 consecutive sections revealed that the other cells were CD68-positive and CD45-positive macrophages and CD31-positive proliferating vascular endothelial cells. We confirmed that PD-L1 was highly expressed in the invasive frontal region in 3D models of MSI-H colon cancer tissue. This method can be useful for accurately evaluating the localisation of immune checkpoint molecules. © 2017 John Wiley & Sons Ltd.

  6. The standard Romanowsky-Giemsa stain in histology.

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    Wittekind, D; Schulte, E; Schmidt, G; Frank, G

    1991-01-01

    A new and technically simple Romanowsky-Giemsa (RG) stain is proposed as a standardized technique for use in histology. An RG stock solution (pure azure B 7.5 g/l, eosin Y as eosinic acid 1.2 g/l in dimethylsulfoxide) is diluted to form the working solution with HEPES-buffer, pH 6. Staining time is 30-90 min after formol-calcium solution (or 2-4 hr after formaldehyde-organic acid mixtures). The resulting overstained sections are to be differentiated. A tannic acid-acetic acid combination in an isopropanol-water mixture was found to give optimum results within 100 sec. Subsequent dehydration is in isopropanol only. The staining pattern obtained is polychrome. The distribution of colors in detail is influenced by the modes of pre- and posttreatment. Of practical interest is the development of green and greenish blue colors on collagen fibrils which contrast strongly against the pink of sarcoplasm. For this and other reasons, this RG stain version seems suitable to replace the trichrome Gomori-type trichrome stains under appropriate processing conditions.

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

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    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.

  8. 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.

  9. 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.

  10. Microwave-stimulated staining of plastic embedded bone marrow sections with the Romanowsky-Giemsa stain: improved staining patterns.

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    Boon, M E; Kok, L P; Moorlag, H E; Gerrits, P O; Suurmeijer, A J

    1987-07-01

    Staining plastic sections with the Romanowsky-Giemsa method is both time-consuming and difficult. This paper reports how the staining time can be reduced to 25 min using microwave irradiation of the staining solution. It is shown that staining results depend on the fixative used, staining temperature, dye concentration and pH of the staining solution as well as on several parameters of the microwave irradiation technique. The staining patterns are improved when compared with those obtained by conventional staining of plastic sections. The colors are more brilliant and greater contrasts are observed. Basophilia, polychromasia, and orthochromasia accompanying red cell maturation are more pronounced. For white cell maturation the initial appearance of specific granules (neutrophil, basophil, and eosinophil) is more evident. Thus, cell classification is easily accomplished using the described technique. It is suggested that microwave-stimulated staining be considered for routine use.

  11. 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.

  12. Real-time histological imaging of kidneys stained with food dyes using multiphoton microscopy.

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    Nagao, Yasuaki; Kimura, Kazushi; Wang, Shujie; Fujiwara, Takeshi; Mizoguchi, Akira

    2015-10-01

    We have developed a real-time imaging technique for diagnosis of kidney diseases which is composed of two steps, staining renal cells safely with food dyes and optical sectioning of living renal tissue to obtain histological images by multiphoton microscopy (MPM). Here, we demonstrated that the MPM imaging with food dyes, including erythrosine and indigo carmine, could be used as fluorescent agents to visualize renal functions and structures such as glomerular bloodstreams, glomerular filtration, and morphology of glomeruli and renal tubules. We also showed that the kidneys of IgA nephropathy model-mice stained with the food dyes presented histopathological characteristics different from those observed in normal kidneys. The use of the food dyes enhances the quality of tissue images obtained by MPM and offers the potential to contribute to a clinical real-time diagnosis of kidney diseases. © 2015 Wiley Periodicals, Inc.

  13. [The staining of decalcified histological bone slices with the improved Romanowsky-Giemsa solution].

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    Drescher, B

    1990-01-01

    This article presents a modified Giemsa-staining. The dye Azure B-Eosinate (Serva, Heidelberg) reformed according to Wittekind was applied to paraffin-slices of decalcified bones of rabbits. Besides a contrast staining on the whole significant details in the histological mounting of bone can be exposed.

  14. A hardware approach for histological and histopathological digital image stain normalization.

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    Şerbănescu, Mircea Sebastian; Pleşea, Iancu Emil

    2015-01-01

    Advances in technology made the migration of pathological diagnosis to digital slides possible. As the need for objectivity and automation emerged, new computer software algorithms were proposed. Computer algorithms demand accurate color and intensity values in order to provide reliable results. The tissue samples undergo several processing steps from histological preparation to digitalization, which cannot be completely standardized. Thus, non-standardized input data generates unreliable output data. In this article, we discuss a new computational normalization algorithm for histopathological stained slides that uses a hardware color marker. The marker is added to the glass slide together with the tissue section, exposed to all the processing steps and altered in the same manner as the biological material of interest, thus becoming a solid color marker for image normalization. The results of the proposed method are numerically and perceptually tested in order to prove the advantages of the method. We conclude that our combined hardware-software technique for staining normalization of digital slides is superior to the existing methods based on only software normalization, and that its implementation will tackle not only the acquisition errors but also the technical errors that may occur during the staining process.

  15. Extracts of Pterocarpus osun as a histological stain for collagen fibres

    African Journals Online (AJOL)

    The staining ability of Pterocarpus osun extract on tissue sections was determined. 2 kg of P. osun stem was dried, milled to obtain a fine powder and a red pigment extracted from the powder with 1 L of 70% ethanol at 78°C for 24 h. The alcoholic and acidic extracts were used to stain tissue sections. Collagen fibres, red ...

  16. Use of Romanowsky type (Diff-3) stain for detecting Helicobacter pylori in smears and tissue sections.

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    Zaitoun, A M

    1992-05-01

    A Romanowsky type (Diff-3) stain was used for identifying Helicobacter pylori in gastric biopsy specimens from 50 patients with ulcer and non-ulcer dyspepsia. Air dried smears were prepared from fresh biopsy tissue and histological sections were prepared from paraffin wax processed tissue. The Diff-3 technique is accomplished in five steps and takes about 30 seconds. Results using the Diff-3 stain correlated 100% with those using the Giemsa stain. The Diff-3 stain is reliable, simple, rapid, easy and clean, and smears prepared from fresh biopsy tissue can be examined and an immediate report given. The method is recommended for the identification of H pylori in smears prepared from fresh tissue as well as in sections prepared from processed tissue.

  17. A contribution to the histological study of hypercementosis using metal staining.

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    Lia, R C; Lauand, F; Marcantonio, E; Neto, C B

    1978-01-01

    After studying hypercementosis, in detail using metallic staining procedures, the authors reached the following conclusions: (1) The histology of hypercementosis, observed by metallic staining reveals remarkable and characteristic microscopical alterations. (2) The Rio Hortega12 procedures, the mitochondria technique of Polak,10 and the method of Achucarro-Hortega13 are of great value for the study of the morphology of cementum. (3) Among the technical procedures used, we consider the 1916 method of Achucarro-Hortega as affording the best view of cementocytes. (4) The Sharpey fibers, revealing great irregularity in course, shape and size, and frequent interruptions could be clearly observed; heavily stained lacunae were frequently observed along the cementum-dentin border; a direct deposit of cellular cementum on the dentin in the middle third of the root was frequently observed. Curiously, an acellular cementum deposit next to a cellular cementum layer could be observed; we also noted numerous cementocytes of different shapes and sizes, at times exhibiting alterations of a degenerative type, particularly fragmentation of protoplasmic prolongations. (5) The techniques used also showed the frequency of blood vessels in the texture of hyperplastic cementum and the presence of capillaries near cementum-dentin junction.

  18. Staining protocol for the histological study of sea anemones (Anthozoa: Actiniaria with recommendations for anesthesia and fixation of specimens

    Directory of Open Access Journals (Sweden)

    Carlos Spano

    2013-11-01

    Full Text Available Many of the characteristics used in sea anemone taxonomy can only be examined through histological sections. Since there is no standardized procedure for this purpose, various anesthesia and fixation techniques applied to specimens of the intertidal species Anthopleura hermaphroditica and Bunodactis hermafroditica are discussed. Additionally, further modifications are proposed to the Masson's trichrome method according to the results obtained on these species. The combined effect of the short application of menthol crystals, together with small doses of MgCl2 were the most satisfactory anesthetics for maintaining the specimens expanded. The best preparations were obtained from samples fixed for several months in 8% seawater formalin; however, in order to achieve a good differentiation of the tissue, mordanting the samples with Bouin's fixative was necessary. Besides being a fast method, the modified Masson's trichrome gives very good contrasts between the epithelia and the mesoglea, and allows controlling the timing of differentiation during staining. The present paper includes suggestions and precautions and thus offers practical help for the histological study of sea anemones.

  19. 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

    The need for the standardization of reagents and methods used in the histology laboratory is demonstrated. After definitions of dyes, stains, and chromogenic reagents, existing standards and standards organizations are discussed. This is followed by practical instructions on how to standardize dyes...

  20. Refractile mycobacteria in Romanowsky-stained bone marrow smears. A comparison of acid-fast-stained tissue sections and Romanowsky-stained smears.

    Science.gov (United States)

    Torlakovic, E; Clayton, F; Ames, E D

    1992-03-01

    The appearance of mycobacteria was studied in Wright-stained bone marrow preparations of human immunodeficiency virus-infected patients and compared with acid-fast-stained trephine biopsy sections and culture results. Mycobacterium avium complex in Romanowsky-stained preparations may be seen as extracellular and intracellular clear or red refractile beaded rods and nonrefractile "negative images." Refractile mycobacteria were seen in 17 of 20 culture-positive cases. Acid-fast stain of the trephine biopsy demonstrated organisms in only 11 of the 20 cases. Thus, six cases were culture positive and contained refractile rods but had no acid-fast organisms on the trephine biopsy. No false-positive results were seen with Romanowsky stain; the three false-negative results for refractility also were negative with acid-fast stain. Examination of Romanowsky-stained smears or imprints for refractile mycobacteria provides a reliable and sensitive method to identify mycobacteria in this population. Romanowsky-stained bone marrow aspirate and imprint smears should be examined for refractile bacilli when mycobacterial infection is suspected.

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

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    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. Buffered Romanowsky-Giemsa method for formalin fixed, paraffin embedded sections: taming a traditional stain.

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    Stefanović, D; Samardžija, G; Redžek, A; Arnaut, M; Nikin, Z; Stefanović, M

    2017-01-01

    Romanowsky-Giemsa (RG) stains were devised during the 19th century for identifying plasmodia parasites in blood smears. Later, RG stains became standard procedures for hematology and cytology. Numerous attempts have been made to apply RG staining to formalin-fixed paraffin-embedded tissue sections, with varied success. Most published work on this topic described RG staining methods in which sections were overstained, then subjected to acid differentiation; unfortunately, the differentiation step often caused inconsistent staining outcomes. If staining is performed under optimal conditions with control of dye concentration, pH, solution temperature and staining time, no differentiation is required. We used RG and 0.002 M buffer, pH 42, for staining and washing sections. All steps were performed at room temperature. After staining and air drying, sections were washed in 96-100% ethanol to remove extraneous stain. Finally, sections were washed in xylene and mounted using DPX. Staining results were similar to routine hemalum and eosin (H & E) staining. Nuclei were blue; intensity depended largely on chromatin density. RNA-rich sites were purple. Collagen fibers, keratin, muscle cells, erythrocytes and white matter of the central nervous system were stained pinkish and reddish hues. Cartilage matrix, mast cell granules and areas of myxomatous degeneration were purple. Sulfate-rich mucins were stained pale blue, while those lacking sulfate groups were unstained. Deposits of hemosiderin, lipofuscin and melanin were greenish, and calcium deposits were blue. Helicobacter pylori bacteria were violet to purple. The advantages of the method are its close similarity to H & E staining and technical simplicity. Hemosiderin, H. pylori, mast cell granules, melanin and specific granules of different hematopoietic cells, which are invisible or barely distinguishable by H & E staining, are visualized. Other advantages over previous RG stains include shorter staining time and avoidance

  3. Comparison of RNA extraction kits and histological stains for laser capture microdissected prostate tissue

    NARCIS (Netherlands)

    K. Kolijn (Kimberley); G.J.H.L. Leenders (Geert)

    2016-01-01

    markdownabstractBackground: Laser capture microdissection offers unique possibilities for the isolation of specific cell populations or histological structures. However, isolation of RNA from microdissected tissue is challenging due to degradation and minimal yield of RNA during laser capture

  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. Method for histological preparation of bone sections containing titanium implants.

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    Hipp, J A; Brunski, J B; Cochran, G V

    1987-07-01

    A thin sectioning technique involving hand grinding has been developed to produce 20-40-microns-thick sections of bone-titanium implant sites. Components include: 1) surface staining of sections prior to mounting on slides so bone labels (oxytetracycline-HCl and 2,4-bis(N,N-dicarbomethyl)aminomethylfluorescein (DCAF] can be seen in sections viewed with transmitted light, 2) a pneumatic sample press for bonding sections to slides with a thin, uniform glue line and without trapped air bubbles, and 3) bonding methyl methacrylate embedded sections to clear acrylic slides with methyl methacrylate monomer to provide enhanced bond strength and grinding properties compared to those obtainable with glass slides. Sample cracking and distortion is minimized and the tissue-implant interface can be kept intact. The expense of start-up equipment for this technique is minimal.

  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. 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.

  8. Immunogold Staining of London Resin (LR) White Sections for Transmission Electron Microscopy (TEM).

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    Skepper, Jeremy N; Powell, Janet M

    2008-06-01

    INTRODUCTIONIn post-embedding methods of immunogold staining, the cells or tissues are fixed chemically or cryoimmobilized, dehydrated, and embedded in epoxy or acrylic resins. Thin sections (50-70 nm in thickness) are cut using an ultramicrotome with a diamond knife, using a water bath to collect the sections as they slide off the knife. The sections are stretched with solvent vapor or a heat source and collected onto either bare or plastic-coated nickel grids. The sections are then stained immunochemically with primary antibodies raised against antigens exposed on the surface of the sections. The primary antibodies are visualized by staining immunochemically with secondary antibodies raised against the species and isotype of the primary antibodies, conjugated to colloidal gold particles. The immunochemically stained sections are then contrast stained with salts of uranium (uranyl acetate) and lead (lead citrate) to reveal the ultrastructure of the cells, and are finally viewed by transmission electron microscopy (TEM). LR White was introduced as a low-toxicity alternative to epoxy resins, which frequently contained carcinogens. Unlike the simplest acrylic resins, in which monomers are polymerized to form long chains, the LR resins contain aromatic cross-linkers to improve the stability of the sections under the electron beam. LR White and Gold both have very low viscosity and readily penetrate, even into dense tissue. In this protocol, aldehyde-fixed tissue is dehydrated in ethanol, impregnated in LR White resin and polymerized under vacuum or in a nitrogen atmosphere before sectioning and immunogold staining.

  9. 3D reconstruction of digitized histological sections for vasculature quantification in the mouse hind limb

    Science.gov (United States)

    Xu, Yiwen; Pickering, J. Geoffrey; Nong, Zengxuan; Gibson, Eli; Ward, Aaron D.

    2014-03-01

    In contrast to imaging modalities such as magnetic resonance imaging and micro computed tomography, digital histology reveals multiple stained tissue features at high resolution (0.25μm/pixel). However, the two-dimensional (2D) nature of histology challenges three-dimensional (3D) quantification and visualization of the different tissue components, cellular structures, and subcellular elements. This limitation is particularly relevant to the vasculature, which has a complex and variable structure within tissues. The objective of this study was to perform a fully automated 3D reconstruction of histology tissue in the mouse hind limb preserving the accurate systemic orientation of the tissues, stained with hematoxylin and immunostained for smooth muscle α actin. We performed a 3D reconstruction using pairwise rigid registrations of 5μm thick, paraffin-embedded serial sections, digitized at 0.25μm/pixel. Each registration was performed using the iterative closest points algorithm on blood vessel landmarks. Landmarks were vessel centroids, determined according to a signed distance map of each pixel to a decision boundary in hue-saturation-value color space; this decision boundary was determined based on manual annotation of a separate training set. Cell nuclei were then automatically extracted and corresponded to refine the vessel landmark registration. Homologous nucleus landmark pairs appearing on not more than two adjacent slides were chosen to avoid registrations which force curved or non-sectionorthogonal structures to be straight and section-orthogonal. The median accumulated target registration errors ± interquartile ranges for the vessel landmark registration, and the nucleus landmark refinement were 43.4+/-42.8μm and 2.9+/-1.7μm, respectively (pextracted vasculature and nuclei.

  10. Automatic segmentation of histological structures in mammary gland tissue sections.

    Science.gov (United States)

    Fernandez-Gonzalez, R; Deschamps, T; Idica, A; Malladi, R; Ortiz de Solorzano, C

    2004-01-01

    Real-time three-dimensional (3-D) reconstruction of epithelial structures in human mammary gland tissue blocks mapped with selected markers would be an extremely helpful tool for diagnosing breast cancer and planning treatment. Besides its clear clinical application, this tool could also shed a great deal of light on the molecular basis of the initiation and progression of breast cancer. We present a framework for real-time segmentation of epithelial structures in two-dimensional (2-D) images of sections of normal and neoplastic mammary gland tissue blocks. Complete 3-D rendering of the tissue can then be done by surface rendering of the structures detected in consecutive sections of the blocks. Paraffin-embedded or frozen tissue blocks are first sliced and sections are stained with hematoxylin and eosin. The sections are then imaged using conventional bright-field microscopy and their background corrected using a phantom image. We then use the fast-marching algorithm to roughly extract the contours of the different morphological structures in the images. The result is then refined with the level-set method, which converges to an accurate (subpixel) solution for the segmentation problem. Finally, our system stacks together the 2-D results obtained in order to reconstruct a 3-D representation of the entire tissue block under study. Our method is illustrated with results from the segmentation of human and mouse mammary gland tissue samples. (c) 2004 Society of Photo-Optical Instrumentation Engineers.

  11. Automatic segmentation of histological structures in mammary gland tissue sections

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Gonzalez, Rodrigo; Deschamps, Thomas; Idica, Adam K.; Malladi, Ravikanth; Ortiz de Solorzano, Carlos

    2004-02-17

    Real-time three-dimensional (3D) reconstruction of epithelial structures in human mammary gland tissue blocks mapped with selected markers would be an extremely helpful tool for breast cancer diagnosis and treatment planning. Besides its clear clinical application, this tool could also shed a great deal of light on the molecular basis of breast cancer initiation and progression. In this paper we present a framework for real-time segmentation of epithelial structures in two-dimensional (2D) images of sections of normal and neoplastic mammary gland tissue blocks. Complete 3D rendering of the tissue can then be done by surface rendering of the structures detected in consecutive sections of the blocks. Paraffin embedded or frozen tissue blocks are first sliced, and sections are stained with Hematoxylin and Eosin. The sections are then imaged using conventional bright field microscopy and their background is corrected using a phantom image. We then use the Fast-Marching algorithm to roughly extract the contours of the different morphological structures in the images. The result is then refined with the Level-Set method which converges to an accurate (sub-pixel) solution for the segmentation problem. Finally, our system stacks together the 2D results obtained in order to reconstruct a 3D representation of the entire tissue block under study. Our method is illustrated with results from the segmentation of human and mouse mammary gland tissue samples.

  12. Segmentation of elastic fibres in images of vessel wall sections stained with Weigert's resorcin-fuchsin.

    Science.gov (United States)

    Hernández-Morera, Pablo; Travieso-González, Carlos M; Castaño-González, Irene; Mompeó-Corredera, Blanca; Ortega-Santana, Francisco

    2017-04-01

    The elastic fibres are an essential component of the extracellular matrix in blood vessel walls that allows a long-range of deformability and passive recoil without energy input. The quantitative determination of elastic fibres will provide information on the state of the vascular wall and to determine the role and behaviour of this key structural element in different physiological and pathological vascular processes. We present a segmentation method to identify and quantify elastic fibres based on a local threshold technique and some morphological characteristics measured on the segmented objects that facilitate the discrimination between elastic fibres and other image components. The morphological characteristics analysed are the thickness and the length of an object. The segmentation method was evaluated using an image database of vein sections stained with Weigert's resorcin-fuchsin. The performance results are based on a ground truth generated manually resulting in values of sensitivity greater than 80% with the exception in two samples, and specificity values above 90% for all samples. Medical specialists carried out a visual evaluation where the observations indicate a general agreement on the segmentation results' visual quality, and the consistency between the methodology proposed and the subjective observation of the doctors for the evaluation of pathological changes in vessel wall. The proposed methodology provides more objective measurements than the qualitative methods traditionally used in the histological analysis, with a significant potential for this method to be used as a diagnostic aid for many other vascular pathological conditions and in similar tissues such as skin and mucous membranes. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. 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...

  14. Automatic segmentation of histological structures in normal and neoplastic mammary gland tissue sections

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Gonzalez, Rodrigo; Deschamps, Thomas; Idica, Adam K.; Malladi, Ravi; Ortiz de Solorzano, Carlos

    2003-01-18

    In this paper we present a scheme for real time segmentation of histological structures in microscopic images of normal and neoplastic mammary gland sections. Paraffin embedded or frozen tissue blocks are sliced, and sections are stained with hematoxylin and eosin (H&E). The sections are then imaged using conventional bright field microscopy. The background of the images is corrected by arithmetic manipulation using a ''phantom.'' Then we use the fast marching method with a speed function that depends on the brightness gradient of the image to obtain a preliminary approximation to the boundaries of the structures of interest within a region of interest (ROI) of the entire section manually selected by the user. We use the result of the fast marching method as the initial condition for the level set motion equation. We run this last method for a few steps and obtain the final result of the segmentation. These results can be connected from section to section to build a three-dimensional reconstruction of the entire tissue block that we are studying.

  15. Automatic segementation of histological structures in normal and neoplastic mammary gland tissue sections

    Science.gov (United States)

    Fernandez-Gonzalez, Rodrigo; Deschamps, Thomas; Idica, Adam; Malladi, Ravikanth; Ortiz de Solorzano, Carlos

    2003-07-01

    In this paper we present a scheme for real time segmentation of histological structures in microscopic images of normal and neoplastic mammary gland sections. Paraffin embedded or frozen tissue blocks are sliced, and sections are stained with hematoxylin and eosin (H&E). The sections are then imaged using conventional bright field microscopy. The background of the images is corrected by arithmetic manipulation using a "phantom." Then we use the fast marching method with a speed function that depends on the brightness gradient of the image to obtain a preliminary approximation to the boundaries of the structures of interest within a region of interest (ROI) of the entire section manually selected by the user. We use the result of the fast marching method as the initial condition for the level set motion equation. We run this last method for a few steps and obtain the final result of the segmentation. These results can be connected from section to section to build a three-dimensional reconstruction of the entire tissue block that we are studying.

  16. Automated classification of inflammation in colon histological sections based on digital microscopy and advanced image analysis.

    Science.gov (United States)

    Ficsor, Levente; Varga, Viktor Sebestyén; Tagscherer, Attila; Tulassay, Zsolt; Molnar, Bela

    2008-03-01

    Automated and quantitative histological analysis can improve diagnostic efficacy in colon sections. Our objective was to develop a parameter set for automated classification of aspecific colitis, ulcerative colitis, and Crohn's disease using digital slides, tissue cytometric parameters, and virtual microscopy. Routinely processed hematoxylin-and-eosin-stained histological sections from specimens that showed normal mucosa (24 cases), aspecific colitis (11 cases), ulcerative colitis (25 cases), and Crohn's disease (9 cases) diagnosed by conventional optical microscopy were scanned and digitized in high resolution (0.24 mum/pixel). Thirty-eight cytometric parameters based on morphometry were determined on cells, glands, and superficial epithelium. Fourteen tissue cytometric parameters based on ratios of tissue compartments were counted as well. Leave-one-out discriminant analysis was used for classification of the samples groups. Cellular morphometric features showed no significant differences in these benign colon alterations. However, gland related morphological differences (Gland Shape) for normal mucosa, ulcerative colitis, and aspecific colitis were found (P parameters showed significant differences (P parameters were the ratio of cell number in glands and in the whole slide, biopsy/gland surface ratio. These differences resulted in 88% overall accuracy in the classification. Crohn's disease could be discriminated only in 56%. Automated virtual microscopy can be used to classify colon mucosa as normal, ulcerative colitis, and aspecific colitis with reasonable accuracy. Further developments of dedicated parameters are necessary to identify Crohn's disease on digital slides. Copyright 2008 International Society for Analytical Cytology.

  17. STAINING SECTIONS OF WATER-MISCIBLE RESINS .1. EFFECTS OF THE MOLECULAR-SIZE OF THE STAIN, AND OF RESIN CROSS-LINKING, ON THE STAINING OF GLYCOL METHACRYLATE EMBEDDED TISSUES

    NARCIS (Netherlands)

    GERRITS, PO; HOROBIN, RW; WRIGHT, DJ

    1990-01-01

    Penetration of hydrophilic acid and basic dyes into sections cut from glycol methacrylate (GMA)-embedded tissues was studied; as were the effects on such staining of superficial coatings of thin layers of GMA. Dye size was a major factor in controlling penetration of resin and staining of tissues.

  18. 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 muscle fibres, such as cross-sectional area, are critical factors that determine the health and function (e.g. quality) of the muscle. However, at this time, quantification of muscle characteristics, especially from haematoxylin and eosin stained slides, is still a manual or semi-automatic process....... This procedure is labour-intensive and time-consuming. In this paper, we have developed and validated an automatic image segmentation algorithm that is not only efficient but also accurate. Our proposed automatic segmentation algorithm for haematoxylin and eosin stained skeletal muscle cross-sections consists...... 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...

  19. 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...

  20. Fully automated registration of vibrational microspectroscopic images in histologically stained tissue sections

    OpenAIRE

    Yang, Chen; Niedieker, Daniel; Gro?er?schkamp, Frederik; Horn, Melanie; Tannapfel, Andrea; Kallenbach-Thieltges, Angela; Gerwert, Klaus; Mosig, Axel

    2015-01-01

    Background In recent years, hyperspectral microscopy techniques such as infrared or Raman microscopy have been applied successfully for diagnostic purposes. In many of the corresponding studies, it is common practice to measure one and the same sample under different types of microscopes. Any joint analysis of the two image modalities requires to overlay the images, so that identical positions in the sample are located at the same coordinate in both images. This step, commonly referred to as ...

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

    OpenAIRE

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

    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 abdominoperineal resection. In addition, 12 cadavers were used to obtain cross-sectional anatomical sections. The images were correlated with the histology and anatomy of the resected rectal specimens as well as with the cross-sectional a...

  2. 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.

  3. 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)

  4. 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

  5. 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.

  6. 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.

  7. 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

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

    Science.gov (United States)

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

    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

  9. Alexandrite Laser for the Treatment of Resistant and Hypertrophic Port Wine Stains: A Clinical, Histological and Histochemical Study.

    Science.gov (United States)

    Grillo, E; González-Muñoz, P; Boixeda, P; Cuevas, A; Vañó, S; Jaén, P

    2016-09-01

    Port wine stains (PWSs) are commonly treated by the pulsed dye laser. However, treatment of hypertrophic or resistant PWSs is a major therapeutic challenge. The long-pulsed Alexandrite laser could be a safe and effective treatment for resistant PWSs, due to an increase depth of penetration of 50-75% over PDL. The aim of this study was to assess the efficacy and safety of a long-pulsed Alexandrite laser in patients with hypertrophic, dark and/or resistant PWSs. Pink pale resistant PWS were excluded from the study. Twenty-one patients (age 20-75 years), phototypes I-IV on the Fitzpatrick scale, with PDL dark resistant PWSs were treated with long-pulsed Alexandrite laser. We excluded high phototypes and PDL pink resistant PWSs. All patients were treated with 3 laser sessions at settings of 3-ms pulse duration, 10-mm spot, 35-55J/cm(2), with cooling (Dynamic Cooling Device 50ms with delay 30ms). Laser sessions were repeated approximately every 2 months. Three dermatologists evaluated treatment effectiveness by means of photographs of the patients before and after laser treatment (scale from 0 to 4). Adverse events were registered. Patient satisfaction was also assessed (scale from 0 to 10). Mean global improvement was rated as 2.28. Long-lasting side effects included minimal scarring after blistering in 1 patient. Mean patient satisfaction was 8.5. Our study concludes that long-pulsed Alexandrite laser was effective for treatment of resistant PWSs, although the therapeutical window is narrow with this treatment. Copyright © 2016 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.

  10. 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

  11. 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

  12. 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

  13. [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.

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

    NARCIS (Netherlands)

    Geffen, G.J. van; Moayeri, N.; Bruhn, J.; Scheffer, G.J.; Chan, V.W.; Groen, G.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

  15. 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.

  16. 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

  17. 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

  18. Correlation Between Ultrasound Imaging, Cross-Sectional Anatomy, and Histology of the Brachial Plexus A Review

    NARCIS (Netherlands)

    van Geffen, Geert J.; Moayeri, Nizar; Bruhn, Joergen; 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

  19. Semi-Automated method of analysis of horizontal histological sections of skin for objective evaluation of fractional devices.

    Science.gov (United States)

    Zelickson, Brian D; Walgrave, Susan E; Al-Arashi, Munir Yahya H; Altshuler, Gregory B; Yaroslavsky, Ilya V; Childs, James J; Cohen, Rich H; Erofeev, Andrei V; Depina, Erminaldo F; Smirnov, Mikhail Z; Kist, David A; Tabatadze, David R

    2009-11-01

    The treatment of skin with fractional devices creates columns of micro-ablation or micro-denaturation depending on the device. Since the geometric profiles of thermal damage depend on the treatment parameters or physical properties of the treated tissue, the size of these columns may vary from a few microns to a few millimeters. For objective evaluation of the damage profiles generated by fractional devices, this report describes an innovative and efficient method of processing and evaluating horizontal sections of skin using a novel software program. Ex vivo porcine skin was treated with the Lux1540/10, Lux1540 Zoom and Lux2940 with 500 optics. Horizontal (radial) sections of biopsies were obtained and processed with H&E and NBTC staining. Digital images of the histologic sections were taken in either transmission or reflection illumination and were processed using the SAFHIR program. NBTC- and H&E-stained horizontal sections of ex vivo skin treated with ablative and non-ablative fractional devices were obtained. Geometric parameters, such as depth, diameter, and width of the coagulated layer (if applicable), and micro-columns of thermal damage, were evaluated using the SAFHIR software. The feasibility of objective comparison of the performance of two different fractional devices was demonstrated. The proposed methodology provides a comprehensive, objective, and efficient approach for the comparison of various fractional devices. Correlation of device settings with the objective dimensions of post-treatment damage profiles serve as a powerful tool for the prediction and modulation of clinical response. Copyright 2009 Wiley-Liss, Inc.

  20. 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.

  1. 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.

  2. 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.

  3. [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

  4. 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......-risk prostate cancer. Two hundred ten patients who awaited curative indented therapy were included. We first performed a standard pathological examination of the LN, followed by an extended pathological examination of the patients who were LN negative in the standard examination. The extended pathological...... indicate that an extended pathological examination of LN will improve the staging of intermediate- and high-risk prostate cancer patients; however, we acknowledge that it is both costly and time consuming. We do not recommend the use of cytokeratin staining in routine staining because...

  5. 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.

  6. 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

  7. 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.

  8. Comparison of pump-probe and hyperspectral imaging in unstained histology sections of pigmented lesions.

    Science.gov (United States)

    Wilson, Jesse W; Robles, Francisco E; Deb, Sanghamitra; Warren, Warren S; Fischer, Martin C

    2017-08-01

    Microscopic variations in melanin composition can be mapped through linear and nonlinear optical responses. Though instrumentation to measure linear attenuation is simple and inexpensive, the nonlinear response provides more degrees of freedom with which to spectroscopically resolve pigments. The objective of this study is to assess differences in imaging melanin contrast by comparing hyperspectral (linear) versus pump-probe (nonlinear) microscopy of unstained histology sections of pigmented lesions. The images and analysis we have presented here show that pump-probe uncovers a greater variation in pigment composition, compared with hyperspectral microscopy, and that the two methods yield complimentary biochemical information.

  9. 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

  10. Adiposoft: automated software for the analysis of white adipose tissue cellularity in histological sections.

    Science.gov (United States)

    Galarraga, Miguel; Campión, Javier; Muñoz-Barrutia, Arrate; Boqué, Noemí; Moreno, Haritz; Martínez, José Alfredo; Milagro, Fermín; Ortiz-de-Solórzano, Carlos

    2012-12-01

    The accurate estimation of the number and size of cells provides relevant information on the kinetics of growth and the physiological status of a given tissue or organ. Here, we present Adiposoft, a fully automated open-source software for the analysis of white adipose tissue cellularity in histological sections. First, we describe the sequence of image analysis routines implemented by the program. Then, we evaluate our software by comparing it with other adipose tissue quantification methods, namely, with the manual analysis of cells in histological sections (used as gold standard) and with the automated analysis of cells in suspension, the most commonly used method. Our results show significant concordance between Adiposoft and the other two methods. We also demonstrate the ability of the proposed method to distinguish the cellular composition of three different rat fat depots. Moreover, we found high correlation and low disagreement between Adiposoft and the manual delineation of cells. We conclude that Adiposoft provides accurate results while considerably reducing the amount of time and effort required for the analysis.

  11. Re-purposing of histological tissue sections for corroborative western blot analysis of hypothalamic metabolic neuropeptide expression following delineation of transactivated structures by Fos immuno-mapping.

    Science.gov (United States)

    Alenazi, Fahaad S H; Ibrahim, Baher A; Briski, Karen P

    2015-04-01

    Fos immunocytochemistry is a valuable anatomical mapping tool for distinguishing cells within complex tissues that undergo genomic activation, but it is seldom paired with corroborative molecular analytical techniques. Due to preparatory requirements that include protein cross-linking for specimen sectioning, histological tissue sections are regarded as unsuitable for those methods. Our studies show that pharmacological activation of the hindbrain energy sensor AMPK by AICAR elicits estradiol (E)-dependent patterns of Fos immunolabeling of hypothalamic metabolic loci. Here, Western blotting was applied to hypothalamic tissue removed from histological sections of E- versus oil (O)-implanted ovariectomized (OVX) female rat brain to measure levels of metabolic transmitters associated with Fos-positive structures. In both E and O rats, AICAR treatment elicited alterations in pro-opiomelanocortin, neuropeptide Y, SF-1, and orexin-A neuropeptide expression that coincided with patterns of Fos labeling of structures containing neurons that synthesize these neurotransmitters, e.g. arcuate and ventromedial nuclei and lateral hypothalamic area. O, but not E animals also exhibited parallel augmentation of tissue corticotropin-releasing hormone neuropeptide levels and paraventricular nucleus Fos staining. Data demonstrate the utility of immunoblot analysis as a follow-through technique to capitalize on Fos mapping of transactivation sites in the brain. Findings that induction of Fos immunoreactivity coincides with adjustments in hypothalamic metabolic neuropeptide expression affirms that this functional indicator reflects changes in neurotransmission in pathways governing metabolic outflow. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. 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.

  13. 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

  14. 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

  15. Statistical evaluation of methods for quantifying gene expression by autoradiography in histological sections

    Directory of Open Access Journals (Sweden)

    Lazic Stanley E

    2009-01-01

    Full Text Available Abstract Background In situ hybridisation (ISH combined with autoradiography is a standard method of measuring the amount of gene expression in histological sections, but the methods used to quantify gene expression in the resulting digital images vary greatly between studies and can potentially give conflicting results. Results The present study examines commonly used methods for analysing ISH images and demonstrates that these methods are not optimal. Image segmentation based on thresholding can be subject to floor-effects and lead to biased results. In addition, including the area of the structure or region of interest in the calculation of gene expression can lead to a large loss of precision and can also introduce bias. Finally, converting grey level pixel intensities to optical densities or units of radioactivity is unnecessary for most applications and can lead to data with poor statistical properties. A modification of an existing method for selecting the structure or region of interest is introduced which performs better than alternative methods in terms of bias and precision. Conclusion Based on these results, suggestions are made to reduce bias, increase precision, and ultimately provide more meaningful results of gene expression data.

  16. Methyl green-pyronin Y staining of nucleic acids: studies on the effects of staining time, dye composition and diffusion rates

    DEFF Research Database (Denmark)

    Prentø, P; Lyon, H O

    2003-01-01

    of hypotheses have been put forward concerning kinetics and binding mechanisms. Using both filter paper models containing DNA, RNA or heparin and histological sections, we have attempted to evaluate the kinetics of staining and the role of staining time for methyl green and pyronin Y by applying the dyes...

  17. Microdissection of stained archival tissue.

    Science.gov (United States)

    Gupta, S K; Douglas-Jones, A G; Morgan, J M

    1997-08-01

    In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns sections are dewaxed, stained with haematoxylin and eosin, dried, covered with Sellotape, and the tissue cut out using a scalpel blade under direct visual control. The method is quick, eliminates problems of operator tremor, preserves the architecture of the micro-dissected tissue (for photographic documentation) and requires no special equipment. The presence of Sellotape and adhesive in the reaction mixture has no detrimental effect on the ability to extract DNA or to perform PCR.

  18. 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…

  19. Triaging borderline/mild dyskaryotic Pap cytology with p16/Ki-67 dual-stained cytology testing: Cross-sectional and longitudinal outcome study

    NARCIS (Netherlands)

    M.H. Uijterwaal (Margot); B.I. Witte (Birgit); F.J. van Kemenade (Folkert); D.C. Rijkaart (Dorien); R. de Ridder (Rogier); J. Berkhof (Johannes); G.A.M.A. Balfoort-Van Der Meij (G. A M A); M.C.G. Bleeker; P.J.F. Snijders (Peter); C.J.L.M. Meijer (Chris J. L.)

    2014-01-01

    textabstractBackground: Women with borderline/mildly dyskaryotic (BMD) cytology smears are currently followed up with repeat testing at 6 and 18 months. The objective of this study is to analyse the cross-sectional and longitudinal performance of p16/Ki-67 dual-stained cytology for the detection of

  20. 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.

  1. Image registration of ex-vivo MRI to sparsely sectioned histology of hippocampal and neocortical temporal lobe specimens.

    Science.gov (United States)

    Goubran, Maged; Crukley, Cathie; de Ribaupierre, Sandrine; Peters, Terence M; Khan, Ali R

    2013-12-01

    Intractable or drug-resistant epilepsy occurs in up to 30% of epilepsy patients, with many of these patients undergoing surgical excision of the affected brain region to achieve seizure control. Recent magnetic resonance imaging (MRI) sequences and analysis techniques have the potential to detect abnormalities not identified with diagnostic MRI protocols. Prospective studies involving pre-operative imaging and collection of surgically-resected tissue provide a unique opportunity for verification and tuning of these image analysis techniques, since direct comparison can be made against histopathology, and can lead to better prediction of surgical outcomes and potentially less invasive procedures. To carry out MRI and histology comparison, spatial correspondence between the MR images and the histology images must be found. Towards this goal, a novel pipeline is presented here for bringing ex-vivo MRI of surgically-resected temporal lobe specimens and digital histology into spatial correspondence. The sparsely-sectioned histology images represent a challenge for 3D reconstruction which we address with a combined 3D and 2D registration algorithm that alternates between slice-based and volume-based registration with the ex-vivo MRI. We evaluated our registration method on specimens resected from patients undergoing anterior temporal lobectomy (N=7) and found our method to have a mean target registration error of 0.76±0.66 and 0.98±0.60 mm for hippocampal and neocortical specimens respectively. This work allows for the spatially-local comparison of histology with post-operative MRI and paves the way for eventual correlation with pre-operative MRI image analysis techniques. © 2013.

  2. 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.

  3. A possible explanation for tissue separation observed in histological sections after regenerative periodontal therapy

    Directory of Open Access Journals (Sweden)

    Firas Kabartai

    2017-01-01

    Full Text Available Introduction: Tissue separation after regenerative periodontal therapy is a common histological observation which is thought to be an artifact. However, several articles have indicated that it can be a serious problem directly related to the weak attachment of the newly formed cementum. The Hypothesis: Tissue separation after periodontal regeneration can be the consequence of losing the fibrous attachment from the cement line between new cementum and the root surface. Evaluation of the Hypothesis: A comparison was made between the cementum–dentin junction, the cement line after root resorption and the cement line after periodontal regeneration because they represent the only means by which the body binds the cementum to root surface. After losing the fibrous attachment from the cement line, the stresses may concentrate at the coronal part of the regenerated tissue leading to the development of tissue separation at that level.

  4. Correlation of nuclear morphometry of breast cancer in histological sections with clinicopathological features and prognosis.

    Science.gov (United States)

    Abdalla, Fathi; Boder, Jamela; Markus, Rabia; Hashmi, Hussein; Buhmeida, Abdelbaset; Collan, Yrjö

    2009-05-01

    The relation of nuclear morphometry measurements with clinicopathological features was evaluated along with prognosis in invasive female breast carcinoma in Libyan patients. Data was compared with corresponding results on Finnish, and Nigerian female breast cancer patients. Histological samples from 131 patients of breast carcinoma were retrospectively studied by computerized nuclear morphometry. In each case, 50 nuclei were measured and the mean nuclear morphometric features were calculated and compared with different clinicopathological features, and patient's survival. There was statistically significant correlation between the mean nuclear area (MNA) and most clinicopathological features, with the strongest association observed for nuclear grade (pparameters can identify the aggressive tumor phenotype and provide significant prognostic information in predicting survival and tumors at risk of progression. The cut-off (71.0 mum(2)) of MNA might be applied as quantitative criterium for Libyan nuclear grading to separate patients into good and poor prognosis groups.

  5. Tanycytic Ependymoma: A Challenging Histological Diagnosis

    Directory of Open Access Journals (Sweden)

    Khaled M. Krisht

    2013-01-01

    Full Text Available Tanycytic ependymoma is a rare form of ependymoma that usually arises in the intramedullary spine. It has a unique histology emphasized by the inconspicuous ependymal pattern of cells and close resemblance to schwannoma and astrocytoma. The authors report a 50-year-old man with a cervical tanycytic ependymoma that was initially thought to be a schwannoma. The frozen histology section showed spindle cells with oval and elongated nuclei with occasional hemosiderin deposits present suggesting a preliminary diagnosis of schwannoma. Immunohistochemical staining of the permanent section revealed strong immunoreactivity for glial fibrillary acidic protein with intermittent S-100 positivity, confirming that the tumor was a tanycytic ependymoma. This underlines the challenges involved in making an accurate diagnosis and demonstrates that careful and detailed histological inspection with immunohistochemical stains and ultrastructural microscopy may be necessary to distinguish tanycytic ependymoma from other neoplasms.

  6. 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…

  7. [Detection of damaged myocardial cells in histological sections by photochemical fluorochroming].

    Science.gov (United States)

    Tsimmerman, V G; Tsellarius, Iu G

    1976-01-01

    The effect of photochemical fluorochromizing used in the method suggested consists in the following: after the exposure of sections to short-wave ultraviolet irradiation the preparations start to fluorescence intensively in the visible region with the long-wave the myocardium of mice, dogs and section materials showed that following photochemical fluorochroming the intensity of lumenescence of damaged cells of the myocardium increased markedly as compared with that of intact cells. The authors recommend to use the described method in diagnosis of early stages of ischemic lesions of the myocardium.

  8. Correlation Among Ultrasound, Cross-Sectional Anatomy, and Histology of the Sciatic Nerve A Review

    NARCIS (Netherlands)

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

    2010-01-01

    Background and Objectives: 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

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

    NARCIS (Netherlands)

    Moayeri, N.; Geffen, G.J. van; Bruhn, J.; Chan, V.W.; Groen, G.J.

    2010-01-01

    BACKGROUND AND OBJECTIVES: 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

  10. Evaluation and Comparison of Frequency of Pulp Stones in Histologic Sections of Severe Chronic Periodontitis Affected Teeth and Healthy Teeth

    Directory of Open Access Journals (Sweden)

    F vaziri

    2017-01-01

    Full Text Available Introduction: Relationship between periodontal infection and pulp pathosis has been debated for many years. This human case control study was performed to evaluate the possible effects of moderate to advanced chronic periodontitis on pulp stone formation and pulpal calcifications. Methods: Current study is an experimental case control study.  Samples included 23 hopeless permanent teeth, affected with periodontitis in the case group and 23 periodontally healthy teeth in the control group, which were extracted for reasons like full denture treatment plan. After extraction and sectioning the apical 2-3 mm of root, teeth were immersed in 10% Formalin solution for 7 days. Then, teeth were decalcified in 5% HNO3 for 10 days. After implementation of tissue preparation steps, sections were made of apical, middle and coronal part of root of each sample. After histological processing, sections were examined by an experienced pathologist under a light microscope. Prescence of pulp stone was investigated in the sections and the results of the two groups were compared with each other. Results: Frequency of pulp stone showed no significant differences between the case and control groups in non of sections. (P-value= 0.730, 0.300, 0.760. Conclusion: Inflammation of periodontium in chronic periodontitis has no influence on pulp stone formation.

  11. Digital image analysis of Ki67 proliferation index in breast cancer using virtual dual staining on whole tissue sections: clinical validation and inter-platform agreement.

    Science.gov (United States)

    Koopman, Timco; Buikema, Henk J; Hollema, Harry; de Bock, Geertruida H; van der Vegt, Bert

    2018-01-18

    The Ki67 proliferation index is a prognostic and predictive marker in breast cancer. Manual scoring is prone to inter- and intra-observer variability. The aims of this study were to clinically validate digital image analysis (DIA) of Ki67 using virtual dual staining (VDS) on whole tissue sections and to assess inter-platform agreement between two independent DIA platforms. Serial whole tissue sections of 154 consecutive invasive breast carcinomas were stained for Ki67 and cytokeratin 8/18 with immunohistochemistry in a clinical setting. Ki67 proliferation index was determined using two independent DIA platforms, implementing VDS to identify tumor tissue. Manual Ki67 score was determined using a standardized manual counting protocol. Inter-observer agreement between manual and DIA scores and inter-platform agreement between both DIA platforms were determined and calculated using Spearman's correlation coefficients. Correlations and agreement were assessed with scatterplots and Bland-Altman plots. Spearman's correlation coefficients were 0.94 (p Ki67 proliferation index in breast cancer, as an alternative to manual scoring of whole sections in clinical practice. Inter-platform agreement between two different DIA platforms was excellent, suggesting vendor-independent clinical implementability.

  12. Clinical and histological characterization of oral pemphigus lesions in patients with skin diseases: a cross sectional study from Sudan

    Science.gov (United States)

    2013-01-01

    Background Pemphigus is a rare group of life-threatening mucocutaneous autoimmune blistering diseases. Frequently, oral lesions precede the cutaneous ones. This study aimed to describe clinical and histological features of oral pemphigus lesions in patients with skin disease has been canceled aged 18 years and above, attending outpatient’s facility of Khartoum Teaching Hospital - Dermatology Clinic, Sudan. In addition, the study aimed to assess the diagnostic significance of routine histolopathology along with immunohistochemical (IHC) examination of formalin-fixed, paraffin-embedded biopsy specimens in patients with oral pemphigus. Methods A cross-sectional hospital-based study was conducted from October 2008 to January 2009. A total of 588 patients with confirmed skin has been canceled disease diagnosis completed an oral examination and a personal interview. Clinical evaluations supported with histopathology were the methods of diagnosis. IHC was used to confirm the diagnosis. Location, size, and pain of oral lesions were used to measure the oral disease activity. Results Twenty-one patients were diagnosed with pemphigus vulgaris (PV), 19 of them (mean age: 43.0; range: 20–72 yrs) presented with oral manifestations. Pemphigus foliaceus was diagnosed in one patient. In PV, female: male ratio was 1.1:1.0. Buccal mucosa was the most commonly affected site. Exclusive oral lesions were detected in 14.2% (3/21). In patients who experienced both skin and oral lesion during their life time, 50.0% (9/18) had oral mucosa as the initial site of involvement, 33.3% (6/18) had skin as the primary site, and simultaneous involvement of both skin and oral mucosa was reported by 5.5% (1/18). Two patients did not provide information regarding the initial site of involvement. Oral lesion activity score was higher in those who reported to live outside Khartoum state, were outdoor workers, had lower education and belonged to Central and Western tribes compared with their

  13. Optical Monte Carlo modeling of a true portwine stain anatomy

    Science.gov (United States)

    Barton, Jennifer K.; Pfefer, T. Joshua; Welch, Ashley J.; Smithies, Derek J.; Nelson, Jerry; van Gemert, Martin J.

    1998-04-01

    A unique Monte Carlo program capable of accommodating an arbitrarily complex geometry was used to determine the energy deposition in a true port wine stain anatomy. Serial histologic sections taken from a biopsy of a dark red, laser therapy resistant stain were digitized and used to create the program input for simulation at wavelengths of 532 and 585 nm. At both wavelengths, the greatest energy deposition occurred in the superficial blood vessels, and subsequently decreased with depth as the laser beam was attenuated. However, more energy was deposited in the epidermis and superficial blood vessels at 532 nm than at 585 nm.

  14. 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...... and analysed with a computerized method. Trabecular bone volume (BV/TV) and connectivity density (CD) were estimated in both modalities, whereas trabecular bone pattern factor (TBPf) was estimated on the histological sections only. Trabecular thickness (Tb.Th), number (Tb.N) and separation (Tb...

  15. 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.

  16. 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

  17. Topographical anatomy of the pronator teres muscle and median nerve: a study using histological sections of human fetuses.

    Science.gov (United States)

    Abe, Shinichi; Tomita, Naomitsu; Yamamoto, Masahito; Sato, Minako; Abe, Hiroshi; Murakami, Gen; Rodríguez-Vázquez, Jose Francisco

    2017-01-01

    The median nerve passes through the humeral and ulnar heads of the pronator teres muscle (PT), although variations such as absence of the ulnar head may exist. We observed histological sections of the upper extremity from 24 embryos and fetuses. In the early stage, the PT extended between the radius and the medial epicondyle of the humerus, but no candidate for the ulnar head was found. In mid-term fetuses, the ulnar margin of the PT was attached to the elbow joint capsule. Moreover, in late-stage fetuses, a small deep part of the PT arose from the thick joint capsule of the humero-ulnar joint near the coronoid process of the ulna. This joint capsule also provided the most proximal origin of the flexor digitorum profundus muscle. Therefore, we considered fetal PT origin from the capsule as a likely candidate for the ulnar head. Consequently, the PT seemed to develop from a single anlage through which the median nerve passed, but later - possibly after birth - a small PT origin from the joint capsule appeared to obtain an aponeurosis connecting the muscle fiber to the ulna. This secondary change in PT morphology might explain the muscle variation seen in adults.

  18. Pleural fluid Gram stain

    Science.gov (United States)

    Gram stain of pleural fluid ... mixing it with a violet stain (called a Gram stain). A laboratory specialist uses a microscope to ... reveals an abnormal collection of pleural fluid. The Gram stain can help identify the bacteria that might ...

  19. High thickness histological sections as alternative to study the three-dimensional microscopic human sub-cortical neuroanatomy.

    Science.gov (United States)

    Alho, Eduardo Joaquim Lopes; Alho, Ana Tereza Di Lorenzo; Grinberg, Lea; Amaro, Edson; Dos Santos, Gláucia Aparecida Bento; da Silva, Rafael Emídio; Neves, Ricardo Caires; Alegro, Maryana; Coelho, Daniel Boari; Teixeira, Manoel Jacobsen; Fonoff, Erich Talamoni; Heinsen, Helmut

    2017-11-01

    Stereotaxy is based on the precise image-guided spatial localization of targets within the human brain. Even with the recent advances in MRI technology, histological examination renders different (and complementary) information of the nervous tissue. Although several maps have been selected as a basis for correlating imaging results with the anatomical locations of sub-cortical structures, technical limitations interfere in a point-to-point correlation between imaging and anatomy due to the lack of precise correction for post-mortem tissue deformations caused by tissue fixation and processing. We present an alternative method to parcellate human brain cytoarchitectural regions, minimizing deformations caused by post-mortem and tissue-processing artifacts and enhancing segmentation by means of modified high thickness histological techniques and registration with MRI of the same specimen and into MNI space (ICBM152). A three-dimensional (3D) histological atlas of the human thalamus, basal ganglia, and basal forebrain cholinergic system is displayed. Structure's segmentations were performed in high-resolution dark-field and light-field microscopy. Bidimensional non-linear registration of the histological slices was followed by 3D registration with in situ MRI of the same subject. Manual and automated registration procedures were adopted and compared. To evaluate the quality of the registration procedures, Dice similarity coefficient and normalized weighted spectral distance were calculated and the results indicate good overlap between registered volumes and a small shape difference between them in both manual and automated registration methods. High thickness high-resolution histological slices in combination with registration to in situ MRI of the same subject provide an effective alternative method to study nuclear boundaries in the human brain, enhancing segmentation and demanding less resources and time for tissue processing than traditional methods.

  20. 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.

  1. New Tetrachromic VOF Stain (Type III-G.S for Normal and Pathological Fish Tissues

    Directory of Open Access Journals (Sweden)

    C Sarasquete

    2005-06-01

    Full Text Available A new VOF Type III-G.S stain was applied to histological sections of different organs and tissues of healthy and pathological larvae, juvenile and adult fish species (Solea senegalensis; Sparus aurata; Diplodus sargo; Pagrus auriga; Argyrosomus regius and Halobatrachus didactylus. In comparison to the original Gutiérrez´VOF stain, more acid dyes of contrasting colours and polychromatic/metachromatic properties were incorporated as essential constituents of the tetrachromic VOF stain. This facilitates the selective staining of different basic tissues and improves the morphological analysis of histochemical approaches of the cell components. The VOF-Type III G.S stain is composed of a mixture of several dyes of varying size and molecular weight (Orange G< acid Fuchsin< Light green

  2. Periodic acid incubation can replace hydrochloric acid hydrolysis and trypsin digestion in immunogold--silver staining of bromodeoxyuridine incorporation in plastic sections and allows the PAS reaction

    NARCIS (Netherlands)

    van de Kant, H. J.; de rooij, D. G.

    1992-01-01

    We have examined the possibility of improving the present methods of detecting bromodeoxyuridine (BrdU) and for combining the PAS reaction with the BrdU detection by means of immunogold-silver staining (IGSS). This was done in testes fixed in Carnoy or Bouin, and in parts of the small intestine

  3. A histochemical study of tissue eosinophilia in oral squamous cell carcinoma using Congo red staining.

    Science.gov (United States)

    Joshi, Priya Shirish; Kaijkar, Manasi S

    2013-11-01

    Tumor associated tissue eosinophilia is believed to play a significant role in the biological behavior of the carcinoma. Eosinophil infiltrate in association with the head and neck squamous cell carcinoma (SCC) have been reviewed from time-to-time. The significance of such an association has been variably thought to be either a potential diagnostic tool for stromal invasion or as a prognostic indicator. We aimed to evaluate the efficacy of Congo red staining to differentiate eosinophils in the inflammatory infiltrate in oral squamous cell carcinoma (OSCC) and whether this eosinophilia is associated with the histologic grading in OSCC. The eosinophil infiltration in hematoxylin and eosin (H and E) and Congo red stained sections of 50 biopsies of OSCC were examined. The eosinophil distribution was quantitatively evaluated in both sections as either diffuse or focal and scored as mild, moderate and severe grades. The average number of eosinophils obtained in OSCC stained by H and E and Congo red were then statistically compared by univariate analysis carried out using Student's t-test. P Congo red stain over H and E stain to differentiate eosinophils was excellent and found to be statistically significant (P Congo red staining showed a high sensitivity in staining eosinophils over routine H and E. This staining technique could therefore provide an adjunct to routine H and E in evaluating eosinophils in dysplasia and OSCC cases.

  4. 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.

  5. 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 ...

  6. Treatment influencing down-staging in EORTC Melanoma Group sentinel node histological protocol compared with complete step-sectioning

    DEFF Research Database (Denmark)

    Riber-Hansen, Rikke; Hastrup, Nina; Clemmensen, Ole

    2012-01-01

    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....

  7. Prognostic Classification of Early Ovarian Cancer Based on very Low Dimensionality Adaptive Texture Feature Vectors from Cell Nuclei from Monolayers and Histological Sections

    Directory of Open Access Journals (Sweden)

    Birgitte Nielsen

    2001-01-01

    Full Text Available In order to study the prognostic value of quantifying the chromatin structure of cell nuclei from patients with early ovarian cancer, low dimensionality adaptive fractal and Gray Level Cooccurrence Matrix texture feature vectors were extracted from nuclei images of monolayers and histological sections. Each light microscopy nucleus image was divided into a peripheral and a central part, representing 30% and 70% of the total area of the nucleus, respectively. Textural features were then extracted from the peripheral and central parts of the nuclei images. The adaptive feature extraction was based on Class Difference Matrices and Class Distance Matrices. These matrices were useful to illustrate the difference in chromatin texture between the good and bad prognosis classes of ovarian samples. Class Difference and Distance Matrices also clearly illustrated the difference in texture between the peripheral and central parts of cell nuclei. Both when working with nuclei images from monolayers and from histological sections it seems useful to extract separate features from the peripheral and central parts of the nuclei images.

  8. 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.

  9. Automated segmentation of atherosclerotic histology based on pattern classification

    Directory of Open Access Journals (Sweden)

    Arna van Engelen

    2013-01-01

    Full Text Available Background: Histology sections provide accurate information on atherosclerotic plaque composition, and are used in various applications. To our knowledge, no automated systems for plaque component segmentation in histology sections currently exist. Materials and Methods: We perform pixel-wise classification of fibrous, lipid, and necrotic tissue in Elastica Von Gieson-stained histology sections, using features based on color channel intensity and local image texture and structure. We compare an approach where we train on independent data to an approach where we train on one or two sections per specimen in order to segment the remaining sections. We evaluate the results on segmentation accuracy in histology, and we use the obtained histology segmentations to train plaque component classification methods in ex vivo Magnetic resonance imaging (MRI and in vivo MRI and computed tomography (CT. Results: In leave-one-specimen-out experiments on 176 histology slices of 13 plaques, a pixel-wise accuracy of 75.7 ± 6.8% was obtained. This increased to 77.6 ± 6.5% when two manually annotated slices of the specimen to be segmented were used for training. Rank correlations of relative component volumes with manually annotated volumes were high in this situation (P = 0.82-0.98. Using the obtained histology segmentations to train plaque component classification methods in ex vivo MRI and in vivo MRI and CT resulted in similar image segmentations for training on the automated histology segmentations as for training on a fully manual ground truth. The size of the lipid-rich necrotic core was significantly smaller when training on fully automated histology segmentations than when manually annotated histology sections were used. This difference was reduced and not statistically significant when one or two slices per section were manually annotated for histology segmentation. Conclusions: Good histology segmentations can be obtained by automated segmentation

  10. Acid-fast stain

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003766.htm Acid-fast stain To use the sharing features on this page, please enable JavaScript. The acid-fast stain is a laboratory test that determines ...

  11. Histologic characteristics of Campylobacter pylori (Helicobacter pylori) mediated gastritis.

    Science.gov (United States)

    Madan, E; Kemp, J; Westblom, T U; Chaffin, J; Foster, A M

    1990-01-01

    One hundred-nineteen specimens were reviewed to determine whether or not there were histologic changes specific for Campylobacter pylori (CP), (Helicobacter pylori) mediated gastritis. Hematoxylin and eosin (H&E), Brown-Hopp, and Wright-Giemsa stained sections were examined independently by two pathologists for (a) the presence of acute cryptitis, (b) percent and degree of crypt involvement, and (c) spectrum of inflammatory cells within the lamina propriae. The amount of mucus was quantified on the Periodic Acid Schift (PAS)-Alcian Blue stain sections. Changes in the character of the mucus were noted by using both the PAS-Alcian Blue and the High Iron Diamine-Alcian Blue. A positive specimen for Campylobacter pylori (CP+), (Helicobacter pylori) was defined as one in which curved or spiral shaped microbes were identified on Wright-Giemsa and Brown-Hopp stain. Seventy-eight specimens were CP+ and 41 CP-. Statistically significant histologic findings included the extent and degree of superficial cryptitis and the preponderance of plasma cells in CP+ cases. These findings confirm aspects seen in an animal model and suggest that there is an histologic pattern consistent with C. pylori (Helicobacter pylori) mediated gastritis.

  12. 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.

  13. Stabilized Romanowsky blood stain.

    Science.gov (United States)

    Gilliland, J W; Dean, W W; Stastny, M; Lubrano, G J

    1979-05-01

    It has been shown that the degradation of thiazine dyes which normally occurs in methanolic solution, as in the case of Romanowsky blood stains, can be prevented by making the solution acidic. In a certain range of acidity, the stain precipitates in the form of monothiazine eosinate, but by making the solution sufficiently acidic, eosin is protonated and the precipitate cannot form. These observations have been used to develop a blood stain which is stable, even at elevated temperatures, for several months. For use the stain is neutralized by a specially formulated fixative solution.

  14. 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.

  15. 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... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures of...

  16. 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.

  17. 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

  18. 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...

  19. Endocervical gram stain

    Science.gov (United States)

    ... no symptoms. Alternative Names Gram stain of cervix; Gram stain of cervical secretions References Marrazzo JM, Apicella MA. Neisseria gonorrhoeae (gonorrhea). In: Bennett JE, Dolin R, Blaser MJ, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, Updated Edition . 8th ...

  20. Dramatic Stained Glass.

    Science.gov (United States)

    Prater, Michael

    2002-01-01

    Describes an art project that is appropriate for students in fifth through twelfth grade in which they create Gothic-style stained-glass windows. Discusses how college students majoring in elementary education created stained-glass windows. Addresses how to adapt this lesson for younger students. (CMK)

  1. 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 ...

  2. 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.

  3. Indirect immunofluorescence staining of cultured neural cells.

    Science.gov (United States)

    Barbierato, Massimo; Argentini, Carla; Skaper, Stephen D

    2012-01-01

    Immunofluorescence is a technique allowing the visualization of a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye such as fluorescein isothiocyanate. There are two major types of immunofluorescence staining methods: (1) direct immunofluorescence staining in which the primary antibody is labeled with fluorescence dye and (2) indirect immunofluorescence staining in which a secondary antibody labeled with fluorochrome is used to recognize a primary antibody. This chapter describes procedures for the application of indirect immunofluorescence staining to neural cells in culture.

  4. Histological Characteristics of Bruises with Different Age

    Science.gov (United States)

    Kostadinova-Petrova, Irena; Mitevska, Elida; Janeska, Biljana

    2017-01-01

    BACKGROUND: In forensics bruises as injuries take an important part in the interpretation of the causes of death. Since activating the inflammatory response of the body in their formation, histological analysis of the bruised tissue can provide data on the determination the time when the injury occurred. AIM: The aim of this study is to compare the histological features of 1-day and 5-days old bruises. MATERIAL AND METHODS: Bruised human skin samples, 1-day old in group A and 5-day-old in group B, obtained at autopsy from individuals who died from a violent death, were analyzed in this study. The qualitative microscopic analysis was performed on serial paraffin sections of tissues stained with Hematoxylin-eosin and Pearls Prussian Blue method, using a light microscope connected to a digital camera. RESULTS: Qualitative histological analysis of the studied group A presented with fresh bruises, less than 24 hours old, showed ruptured smaller vessels and extravasated red blood cells in the connective tissue of the skin, with subsequent expansion and infiltration of fibrous septa of the skin. In the area of bleeding an initial infiltration by macrophages was observed. In the studied group B, presented with bruises 3-7 days old, histological analysis showed a marked presence of hemosiderin-laden macrophages and presence of hematoidin granules in the area of bleeding, as well as ruptured small blood vessels and red blood cells extravasation in the dilated fibrous septa. CONCLUSION: A detailed analysis of tissue changes in bruises every day from the initiation until their recovery, a detailed description of the histological finding can be given, which will be supported in the precise determination of the age of the injuries themselves. PMID:29362602

  5. Stool Gram stain

    Science.gov (United States)

    ... of stool; Feces Gram stain References Allos BM. Campylobacter infections. In: Goldman L, Schafer AI, eds. Goldman- ... Bacterial Infections Read more Foodborne Illness Read more Gastroenteritis Read more A.D.A.M., Inc. is ...

  6. Myeloperoxidase staining in the diagnosis of aggressive periodontitis.

    Science.gov (United States)

    Singh, Sukhdeep; Acharya, Anirudh B; Kumar, S C Veerendra

    2011-04-01

    To evaluate neutrophil myeloperoxidase (MPO) staining procedure as a reliable, affordable and easily available diagnostic assay for aggressive periodontitis. Fifteen subjects were recruited in the study wherein five each were diagnosed as aggressive periodontitis and chronic periodontitis respectively, and five were periodontally healthy. Three millilitres (ml) of venous blood was collected using Vacutainers containing ethylene diamine tetra acetate (EDTA) and was subjected to MPO staining procedure. Histological picture was evaluated using a visual analogue scale (VAS). MPO stained specimen of all the patients showed positive MPO staining of the neutrophils. The intensity of the stain of MPO granules was more in aggressive periodontitis specimen as compared to the chronic periodontitis patient specimen and healthy subject specimen. The staining characteristics were comparable for chronic periodontitis patients and healthy subject. This study shows that there is a potential and probable place for MPO staining as an economical, relatively convenient and easily available assay in the diagnosis of aggressive periodontitis.

  7. 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

  8. "Stained Glass" Landscape Windows

    Science.gov (United States)

    Vannata, Janine

    2008-01-01

    Both adults and children alike marvel at the grand vivid stained-glass windows created by American artist Louis Comfort Tiffany. Today he is commonly recognized as one of America's most influential designers and artists throughout the last nineteenth and early twentieth century. In the lesson described in this article, students created their own…

  9. 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.

  10. 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/ ...

  11. Histology of the Ovary of the Laying Hen (Gallus domesticus).

    Science.gov (United States)

    Apperson, K Denise; Bird, Karyn E; Cherian, Gita; Löhr, Christiane V

    2017-12-11

    The laying hen ( Gallus domesticus ) is a robust animal model for epithelial ovarian cancer. The use of animal models is critical in identifying early disease markers and developing and testing chemotherapies. We describe the microscopic characteristics of the normally functioning laying hen ovary and proximal oviduct to establish baselines from which lesions associated with ovarian cancer can be more readily identified. Ovaries and oviducts were collected from 18-month-old laying hens ( n = 18) and fixed in 10% neutral buffered formalin. Hematoxylin- and eosin-stained sections were examined by light microscopy. Both post-ovulatory follicular regression and atresia of small follicles produce remnant clusters of vacuolated cells with no histological evidence that scar tissue persists. Infiltrates of heterophils are associated with atresia of small follicles, a relationship not previously documented in laying hen ovaries. Because these tissues can be mistaken for cancerous lesions, we present a detailed histological description of remnant Wolffian tissues in the laying hen ovary. Immunohistochemical staining for pancytokeratin produced a positive response in ovarian surface epithelium and staining for vimentin produced a positive response in granulosa cells of follicles. Epithelial cells lining glands of the remnant epoöphoron had a positive response to both pancytokeratin and vimentin, a result also observed in women.

  12. Immunoperoxidase staining for involucrin: a potential diagnostic aid in cervicovaginal pathology.

    Science.gov (United States)

    Warhol, M J; Antonioli, D A; Pinkus, G S; Burke, L; Rice, R H

    1982-12-01

    Involucrin, a protein subunit of keratinocyte cross-linked envelopes, is a distinctive marker for suprabasal differentiation in stratified squamous epithelium. Immunoperoxidase staining for involucrin was used to evaluate paraffin sections of tissue obtained by colposcopically directed biopsies of infectious, metaplastic, and dysplastic lesions of the cervix and vagina. Areas of normal squamous epithelium, papillary and flat condyloma acuminatum, and mature and immature squamous metaplasia showed positive staining in 99 per cent of samples lacking significant inflammation and in 60 per cent of those with moderate or severe inflammation. In contrast, only 19 per cent of the squamous cell dysplasias, even those without much inflammation, showed positive staining, and no area with moderate or severe inflammation showed positive staining. These findings indicate that expression of involucrin is modulated by cellular pathologic features and microenvironment. We suggest that immunoperoxidase staining for involucrin may be useful in distinguishing mild dysplasia from immature metaplasia and flat condyloma in some biopsy specimens in which routine histologic examination yields an indeterminate diagnosis.

  13. 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 ...

  14. 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] ...

  15. 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

  16. A comparison of different embalming fluids on the quality of histological preservation in human cadavers.

    Science.gov (United States)

    Nicholson, H D; Samalia, L; Gould, M; Hurst, P R; Woodroffe, M

    2005-01-01

    There are significant problems in obtaining normal human material for histology for teaching or research purposes. This study shows that tissue from cadavers embalmed for teaching can be used for routine histology. Twelve cadavers embalmed with four different formalin-containing embalming fluids were used (n = 3 per fluid): (1) formalin mix (10% formalin); (2) Dunedin mix (an alcohol-based fluid containing phenol); (3) Michigan mix (a water-based fluid); and (4) phenoxyethanol mix (an alcohol-based fluid containing phenoxyethanol). Tissue blocks of liver, heart, kidney, skin and skeletal muscle were taken from each cadaver, paraffin embedded, sectioned and stained with haematoxylin and eosin (H & E), Periodic Acid Schiff (PAS), or Mallory trichrome (Malt). Each section was assigned an overall score based on the histological quality of the cellular components of the tissue. Sections were scored from 1 to 3 (1 = poor, 2 = satisfactory, 3 = good). Satisfactory sections were obtained from all cadavers except those embalmed with the Dunedin mix. The Michigan and phenoxyethanol fluids resulted in consistently good quality sections. No significant differences in tissue morphology were observed between the different stains. The clearest morphology was observed in the skin and skeletal muscle sections, and in tissues embalmed with fluids which do not contain phenol.

  17. 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.

  18. Image analysis of vocal fold histology

    Science.gov (United States)

    Reinisch, Lou; Garrett, C. Gaelyn

    2001-05-01

    To visualize the concentration gradients of collagen, elastin and ground substance in histologic sections of vocal folds, an image enhancement scheme was devised. Slides stained with Movat's solution were viewed on a light microscope. The image was digitally photographed. Using commercially available software, all pixels within a color range are selected from the mucosa presented on the image. Using the Movat's pentachrome stain, yellow to yellow-brown pixels represented mature collagen, blue to blue-green pixels represented young collagen (collagen that is not fully cross-linked) and black to dark violet pixels represented elastin. From each of the color range selections, a black and white image was created. The pixels not within the color range were black. The selected pixels within the color range were white. The image was averaged and smoothed to produce 256 levels of gray with less spatial resolution. This new grey-scale image showed the concentration gradient. These images were further enhanced with contour lines surrounding equivalent levels of gray. This technique is helpful to compare the micro-anatomy of the vocal folds. For instance, we find large concentration of the collagen deep in the mucosa and adjacent to the vocalis muscle.

  19. Usefulness of automatic quantification of immunochemical staining on whole tumor sections for correlation with oncological small animal PET studies: an example with cell proliferation, glucose transporter 1 and FDG.

    Science.gov (United States)

    Aide, Nicolas; Labiche, Alexandre; Herlin, Paulette; Paciencia, Maria; Poulain, Laurent; Dutoit, Soizic; Montravers, Françoise; Gauduchon, Pascal; Chasle, Jacques

    2008-09-01

    To highlight the use of automatic quantification of immunochemical staining on digitized images of whole tumor sections in preclinical positron emission tomography (PET) studies. Xenografted human testicular tumors (36) were imaged with 2-deoxy-2[F-18]fluoro-D: -glucose (FDG) small animal PET (SA-PET). Tumor cell proliferation and glucose transportation were assessed with cyclin A and Glut-1 immunostaining. Tumor slides were digitized and processed with PixCyt software enabling whole slide quantification, then compared with junior and senior pathologist manual scoring. Manual and automatic quantification results were correlated to FDG uptake. For cyclin A, inter- and intra-observer agreement for manual scoring was 0.52 and 0.72 and concordance between senior pathologist and automatic quantification was 0.84. Correlations between Tumor/Background ratio and tumor cell proliferation assessed by automatic quantification, junior and senior pathologists were 0.75, 0.55, and 0.61, respectively. Correlation between Tumor/Background ratio and Glut-1 assessed by automatic quantification was 0.74. Automatic quantification of immunostaining is a valuable tool to overcome inter- and intra-observer variability for correlation of cell proliferation or other markers with tumor tracer uptake.

  20. 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

  1. 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

  2. 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.

  3. Advantage of affinity histochemistry combined with histology to investigate death causes: indications from sample cases.

    Science.gov (United States)

    Bacci, Stefano; DeFraia, Beatrice; Romagnoli, Paolo; Bonelli, Aurelio

    2011-11-01

    Mast cell histochemistry has been proposed in addition to classic histological methods to estimate the course of traumatic events before and after death. We have addressed the utility of this approach on nine victims of different types of trauma. Sections of wounded skin were stained with hematoxylin and eosin and with fluorescent avidin to tag mast cells. Mast cell numbers were evaluated by both direct and digitalized counts. Intact skin was used as control. The results on mast cells implemented the findings upon hematoxylin and eosin stain and helped to put the wounds and death in chronological sequence. Digitalized morphometry allowed to reduce intra- and inter-observer variation. We conclude that combined histological and histochemical analyses can be of practical use in forensic pathology, that a preliminary setting of the reference values is needed for each laboratory, and that image analysis can be of help for the quantification of the results. © 2011 American Academy of Forensic Sciences.

  4. Ghost mycobacteria on Gram stain.

    Science.gov (United States)

    Trifiro, S; Bourgault, A M; Lebel, F; René, P

    1990-01-01

    The Gram stain is a key tool in diagnostic microbiology. Its usefulness with respect to mycobacteria is undefined. The neutrality of mycobacteria other than Mycobacterium tuberculosis on Gram staining of various clinical specimens is described. Images PMID:1688872

  5. Ghost mycobacteria on Gram stain.

    OpenAIRE

    Trifiro, S; Bourgault, A M; Lebel, F; René, P

    1990-01-01

    The Gram stain is a key tool in diagnostic microbiology. Its usefulness with respect to mycobacteria is undefined. The neutrality of mycobacteria other than Mycobacterium tuberculosis on Gram staining of various clinical specimens is described.

  6. Gram stain of urethral discharge

    Science.gov (United States)

    Urethral discharge Gram stain; Urethritis - Gram stain ... Augenbraun MH, McCormack WM. Urethritis. In: Bennett JE, Dolin R, Blaser MJ, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, Updated Edition . ...

  7. Effects of phyto-oestrogens on veal calf prostate histology.

    Science.gov (United States)

    Groot, M J

    2006-08-01

    In veal calf production plant-based proteins are frequently included in milk replacer fed to the animals. Since soy products, which are mostly used, are known for their high levels of phyto-oestrogens, the effects of these feeds on the veal calf prostate were examined. Goal was to determine whether these compounds could interfere with histological screening for oestrogenic growth promoters. In a feeding experiment, four groups of veal calves fed plant-based protein-supplemented milk replacer (PBM), containing 5% soy concentrate, 5% soy isolate, 5% wheat gluten and 2% potato protein, for 4 weeks were compared to animals fed dairy-based control feed (DBM); animals treated with estradiol benzoate, diethylstilbestrol and ethinylestradiol served as positive controls. Daidzein and genistein levels measured in feed and urine showed high levels of genistein and daidzein in the soy isolate and soy concentrate supplemented feeds. Genistein and daidzein were also found in the urine of the animals that were fed these feeds. Haematoxylin-eosin-stained prostate sections of PBM-fed animals showed slight hyperplasia and some dilated tubules as compared to the DBM-fed group, but no metaplasia, which is used for screening for oestrogenic hormones. The positive controls showed extensive squamous metaplasia. Immunohistochemical staining for cytokeratin 5 (using RCK 103 monoclonal antibody) in basal cells showed a normal staining pattern of basal cells in the DBM-fed calves and extensive basal cell proliferation and squamous metaplasia in the oestrogen-treated positive control animals. PBM-fed calves showed no increase of basal cell staining but showed elongations of the basal cells in most animals, sometimes resulting in circular figures. It is concluded that the feeds examined in this study did not interfere with histological screening for oestrogens in male veal calves.

  8. Quantitative three-dimensional evaluation of immunofluorescence staining for large whole mount spheroids with light sheet microscopy.

    Science.gov (United States)

    Smyrek, I; Stelzer, E H K

    2017-02-01

    Three-dimensional cell biology and histology of tissue sections strongly benefit from advanced light microscopy and optimized staining procedures to gather the full three-dimensional information. In particular, the combination of optical clearing with light sheet-based fluorescence microscopy simplifies fast high-quality imaging of thick biological specimens. However, verified in toto immunostaining protocols for large multicellular spheroids or for tissue sections have not been published. We present a method for the verification of immunostaining in three-dimensional spheroids. The analysis relies on three criteria to evaluate the immunostaining quality: quality of the antibody stain specificity, signal intensity achieved by the staining procedure and the correlation of the signal intensity with that of a homogeneously dispersed fluorescent dye. We optimized and investigated variations of five immunostaining protocols for three-dimensional cell biology. Our method is an important contribution to three-dimensional cell biology and the histology of tissues since it allows to evaluate the efficiency of immunostaining protocols for large three-dimensional specimens, and to study the distribution of protein expression and cell types within spheroids and spheroid-specific morphological structures without the need of physical sectioning.

  9. Occult cancer with cervical lymph node metastasis: histologic profile ...

    African Journals Online (AJOL)

    endoscopy biopsy specimen was appreciable. Inadequate immunohistological stains and lack of FDG PET scan may have diminished the histological yield of the blind pan-endoscopy biopsy specimens. Keywords: Cervical nodal metastasis ...

  10. 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.

  11. 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.

  12. 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.

  13. Stain-free histopathology by programmable supercontinuum pulses

    Science.gov (United States)

    Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry; Marjanovic, Marina; Lyngsø, Jens K.; Lægsgaard, Jesper; Chaney, Eric J.; Zhao, Youbo; You, Sixian; Wilson, William L.; Xu, Bingwei; Dantus, Marcos; Boppart, Stephen A.

    2016-08-01

    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-intensive. Here, we introduce a unique optical imaging platform and methodology for label-free multimodal multiphoton microscopy that uses a novel photonic-crystal fibre source to generate tailored chemical contrast based on programmable supercontinuum pulses. We demonstrate the collection of optical signatures of the tumour microenvironment, including evidence of mesoscopic biological organization, tumour cell migration and (lymph-) angiogenesis collected directly from fresh ex vivo mammary tissue. Acquisition of these optical signatures and other cellular or extracellular features, which are largely absent from histologically processed and stained tissue, combined with an adaptable platform for optical alignment-free programmable-contrast imaging, offers the potential to translate stain-free molecular histopathology into routine clinical use.

  14. Toluidine Blue 0.05% Vital Staining for the Diagnosis of Ocular Surface Squamous Neoplasia in Kenya.

    Science.gov (United States)

    Gichuhi, Stephen; Macharia, Ephantus; Kabiru, Joy; Zindamoyen, Alain M'bongo; Rono, Hilary; Ollando, Ernest; Wanyonyi, Leonard; Wachira, Joseph; Munene, Rhoda; Onyuma, Timothy; Jaoko, Walter G; Sagoo, Mandeep S; Weiss, Helen A; Burton, Matthew J

    2015-11-01

    Clinical features are unreliable for distinguishing ocular surface squamous neoplasia (OSSN) from benign conjunctival lesions. To evaluate the adverse effects, accuracy, and interobserver variation of toluidine blue 0.05% vital staining in distinguishing OSSN, confirmed by histopathology, from other conjunctival lesions. Cross-sectional study in Kenya from July 2012 through July 2014 of 419 adults with suspicious conjunctival lesions. Pregnant and breastfeeding women were excluded. Comprehensive ophthalmic slitlamp examination was conducted. Vital staining with toluidine blue 0.05% aqueous solution was performed before surgery. Initial safety testing was conducted on large tumors scheduled for exenteration looking for corneal toxicity on histology before testing smaller tumors. We asked about pain or discomfort after staining and evaluated the cornea at the slitlamp for epithelial defects. Lesions were photographed before and after staining. Diagnosis was confirmed by histopathology. Six examiners assessed photographs from a subset of 100 consecutive participants for staining and made a diagnosis of OSSN vs non-OSSN. Staining was compared with histopathology to estimate sensitivity, specificity, and predictive values. Adverse effects were enumerated. Interobserver agreement was estimated using the κ statistic. A total of 143 of 419 participants (34%) had OSSN by histopathology. The median age of all participants was 37 years (interquartile range, 32-45 years) and 278 (66%) were female. A total of 322 of the 419 participants had positive staining while 2 of 419 were equivocal. There was no histological evidence of corneal toxicity. Mild discomfort was reported by 88 (21%) and mild superficial punctate keratopathy seen in 7 (1.7%). For detecting OSSN, toluidine blue had a sensitivity of 92% (95% CI, 87%-96%), specificity of 31% (95% CI, 25%-36%), positive predictive value of 41% (95% CI, 35%-46%), and negative predictive value of 88% (95% CI, 80%-94%). Interobserver

  15. 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.

  16. Staining properties and stability of a standardised Romanowsky stain.

    Science.gov (United States)

    Marshall, P N; Bentley, S A; Lewis, S M

    1978-03-01

    An evaluation of the standardised Romanowsky stain of Marshall et al. has been made in a routine haematology laboratory. It was noted that this stain had several advantages over the May-Grünwald Giemsa stain used in most British laboratories. These advantages include ease and speed of preparation, a shorter staining time, and reproducibility of results. These results are described in detail. The stability of the stock stain solution and of the 'working' stain (stock + buffer) has been studied by, respectively, thin-layer chromatography and visible spectroscopy. No change was detected in the composition of the stock solution at ambient temperature over a period of six months. Stability was unaffected by the composition of the container (polyethylene, PyrexTM, or soda-glass) or by daylight. The 'working' solution was stable for 3 hours. Thereafter a precipitate is formed, consisting of thiazine dyes and eosin in a molar ratio of approximately 2:1.

  17. 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.

  18. Routine histologic demonstration of helicobacter pylori in gastric ...

    African Journals Online (AJOL)

    Routine histologic demonstration of helicobacter pylori in gastric biopsies: should immunohistochemistry replace giemsa stain? ... Method: Thirty-five consecutive endoscopic gastric biopsies obtained from dyspeptic patients, having a histological diagnosis of chronic gastritis were reviewed. Giemsa and IHC were applied ...

  19. Biological, histological and ultra-structural studies of female mullet ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-07-29

    Jul 29, 2015 ... This study investigates some biological characters (oocyte diameter, fecundity, histological and ultra structural features) of ... histological and fine structure characteristics, it was possible to understand the functional relationship between .... thick were prepared using glass and diamond knives, and stained.

  20. Histological effect of ethanolic leaf extract of Codiaeum variegatum ...

    African Journals Online (AJOL)

    Histological effect of ethanolic leaf extract of Codiaeum variegatum on the cerebrum of adult Wistar rats. ... Journal of Experimental and Clinical Anatomy ... They were sacrificed on the 15th day of the experiment; cerebrum was harvested, processed, and stained using the hematoxylin and eosin histological technique.

  1. A novel approach to the histological diagnosis of pediatric nephrotic syndrome by low vacuum scanning electron microscopy.

    Science.gov (United States)

    Okada, Shinichi; Inaga, Sumire; Kawaba, Yasuo; Hanada, Takuya; Hayashi, Atsushi; Nakane, Hironobu; Naguro, Tomonori; Kaidoh, Toshiyuki; Kanzaki, Susumu

    2014-01-01

    Despite intensive treatment, steroid-resistant nephrotic syndrome (NS) often progresses to endstage renal disease. Therefore, a more accurate and quick histological diagnosis is required to properly treat such patients. The aim of this study was to introduce a novel approach to the histological diagnosis of pediatric NS by low vacuum scanning electron microscopy (LVSEM) and to describe the morphological differences in glomeruli between steroid-sensitive and steroid-resistant NS specimens. The subjects were three patients with steroid-sensitive NS and four patients with steroid-resistant NS. Conventional renal biopsy paraffin sections were stained with platinum-blue (Pt-blue) or periodic acid methenamine silver (PAM) and directly observed under LVSEM at magnifications between ×50 and ×10,000. The Pt-blue-stained sections showed three-dimensional structural alterations in glomerular podocytes and foot processes. PAM-stained sections showed changes in the structure and thickness of the glomerular basement membrane (GBM). Consequently, many round-shaped podocytes and elongated primary foot processes were exclusively recognized in steroid-resistant NS, although irregularities in foot process interdigitation, fusions, effacements, and microvillus transformations were observed in both steroid-sensitive and steroidresistant NS. Irregularities in thickness and the wrinkling of GBMs were clearly detected in steroid-resistant NS. The evaluation by LVSEM is probably useful for the renal histological diagnosis of pediatric NS.

  2. Micro-computed tomography scan and virtual histological slide data for the land planarian Obama otavioi (Platyhelminthes).

    Science.gov (United States)

    Carbayo, Fernando; Lenihan, Jennifer Winifred

    2016-01-01

    We investigated whether images obtained through X-ray micro-computed tomography (μCT) can be used in conjunction with traditional methods for morphological studies of soft-bodied land planarians. μCT is non-invasive and provides true-to-scale three-dimensional imagery at high resolution. We compared μCT-based images of a recently described land planarian species of Obama otavioi (Platyhelminthes) with those obtained from light microphotography of histological sections, most of which were also digitized at high magnification. The specimens studied were collected in 2012. Subsequent μCT-based images of the stained body of a paratype show nearly all morphological features provided by traditional histology, with the exception of particularly minute structures, smaller than 5 μm, such as the sensory pits and single muscle fibers, which are best visible on traditional histological sections. Because the technique is non-destructive, the scanned specimen is preserved without damage. The raw and derivative μCT data and virtual histological sections are freely available in GigaDB. The μCT datasets of these stained soft-bodied organisms reveal images of external and internal structures that support previous taxonomic studies. This technique can be particularly important for non-destructively revealing internal details of whole museum specimens at a faster rate than histology alone. High-resolution virtual histological slides also allow further searches for new, previously unstudied morphological features. The use of X-ray equipment with higher resolution can enable smaller sensory organ and muscle fiber details to be seen. The image sets, μCT-based images and digitized histological slides can be disseminated without the constraints of specimen loans.

  3. Morin Stain Detects Aluminum-Containing Macrophages in Macrophagic Myofasciitis and Vaccination Granuloma With High Sensitivity and Specificity.

    Science.gov (United States)

    Chkheidze, Rati; Burns, Dennis K; White, Charles L; Castro, Diana; Fuller, Julie; Cai, Chunyu

    2017-04-01

    Macrophagic myofasciitis (MMF) is an inflammatory condition associated with the intramuscular (i.m.) injection of aluminum adjuvant-containing vaccines. It is clinically characterized by myalgia, weakness, and chronic fatigue and histologically by aggregates of cohesive macrophages with abundant basophilic, periodic acid-Schiff (PAS)-positive, diastase-resistant granules that percolate through the peri- and endomysium without eliciting substantial myofiber damage. The definitive diagnosis of MMF requires demonstration of aluminum within these macrophages. We evaluated the Morin stain, a simple, 2-step histochemical stain for aluminum, as a confirmatory diagnostic tool for MMF. Among 2270 muscle biopsies processed at UTSW between 2010 and 2015, a total of 12 MMF cases and 1 subcutaneous vaccination granuloma case were identified (11 pediatric, 2 adults). With the Morin stain, all 13 cases showed strong granular reactivity within the cytoplasm of macrophages but not in myofibers or connective tissue. Three cases of inflammatory myopathy with abundant macrophages (IMAM), 8 cases of granulomatous inflammation and 23 other deltoid muscle biopsies used as controls were all negative. Morin stain could be used in both formalin-fixed paraffin-embedded and cryostat sections. Thus, Morin stain detects aluminum with high sensitivity and specificity in human muscle and soft tissue and may improve the diagnostic yield of MMF and vaccination granuloma. © 2017 American Association of Neuropathologists, Inc.

  4. 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.

  5. A new bacterial staining method involving Gram stain with theoretical considerations of the staining mechanism.

    Science.gov (United States)

    Noda, Y; Tôei, K

    1992-01-01

    In order to investigate the mechanism of Gram staining of bacteria, tests with anionic dyes followed by treatment with cationic octyltrimethylammonium (OTMA) were carried out. The study revealed that tetrabromophenolphthalein ethylester (TBPE) gave the most reliable staining of Gram-negative bacteria with negative staining of Gram-positive bacteria. Tests on many species of bacteria showed that TBPE positive bacteria were Gram-negative and vice versa, without exception.

  6. Histological underestimation of a 9-gauge stereotactic vacuum ...

    African Journals Online (AJOL)

    Method: In this cross-sectional study, 9-gauge stereotactic biopsy histology results and breast imaging and reporting data system (BI-RADS) findings of 158 lesions (from 153 patients) were analysed and the histological findings compared with surgical excision histology results (54 lesions) to determine histological ...

  7. Histological changes of the adult albino rats entorhinal cortex under ...

    African Journals Online (AJOL)

    For light microscopy, the brains of 10 animals in each group were processed for paraffin sections and stained by Gallocyanine stain. For electron microscopy, the entorhinal cortex was dissected in 10 brains of each group and processed. Semithin sections were prepared and stained with toluidine blue. Morphometric and ...

  8. Characteristics at haematoxylin and eosin staining of ruptures of the long head of the biceps tendon.

    Science.gov (United States)

    Longo, U G; Franceschi, F; Ruzzini, L; Rabitti, C; Morini, S; Maffulli, N; Denaro, V

    2009-08-01

    To examine the relative prevalence of histological changes that have been found to be associated with the process of tendinopathy in lesions of the tendon of the long head of the biceps brachii and to evaluate the reliability of histopathological evaluation of tendon tissue in lesions of the tendon of the long head of the biceps. Tendon samples were taken from 51 patients (31 men, 20 women; mean age 63.2 years) who underwent arthroscopic release of the long head of the biceps tendon because of refractory biceps tendinopathy and from 5 male patients who died of cardiovascular events (mean age 69.6 years). Histological examination was performed using haematoxylin and eosin staining of sections, which were interpreted using a semiquantitative grading scale assessing fibre structure and arrangement, rounding of the nuclei, regional variations in cellularity, increased vascularity, decreased collagen staining and hyalinisation. The mean (SD) pathological sum score of ruptured tendons was greater than that of control tendons (15.76 (3.11) vs 3.4 (1.9), pruptured tendons; all the variables were significantly different (Mann-Whitney U test 0.05, pbiceps, even at an advanced age and ruptured tendons of the long head of the biceps are clearly part of two distinct populations.

  9. An evaluation of some commerical Romanowsky stains.

    Science.gov (United States)

    Marshall, P N; Bentley, S A; Lewis, S M

    1975-08-01

    The staining properties of 43 commerical Romanowsky-type stains have been studied. Considerable differences in the appearance of stained blood films were observed with different batches of these stains, the staining of red cells being particularly variable. Attempts have been made to correlate staining patterns with stain composition as revealed by thin-layer chromatography and sulphated ash analyses. In this way it has been possible to define some essential requirements for satisfactory staining.

  10. Possibly Active Persistent Thymus Found in a Human Adult Cadaver – A Morpho-histological Study

    Directory of Open Access Journals (Sweden)

    Nayak SB

    2016-01-01

    Full Text Available Thymus is a bilobed organ usually situated in the superior mediastinum. Thymus is normally active until puberty and as age advances it undergoes considerable fibro-fatty degeneration and is replaced by fatty tissue. We found a persistent thymus in an adult male cadaver aged 70 years approximately. It apparently looked healthy. Hence the objective of this study was to know the morpho-histology of a persistent human thymus gland. Associated with this we also found a concurrent absence of isthmus of thyroid gland. Thymus obtained was processed according to the standard procedures and sections were stained with Haematoxylin & Eosin stain to study the age related changes of the thymus gland. Stained sections of thymus revealed a normal, healthy architecture of lobes of thymus. Knowledge regarding these morpho-histological features of the persistent thymus is important for clinicians and radiologists for the differential diagnosis of any mediastinal mass or presence of ectopic thymic tissue before doing any investigative procedure

  11. Association between human papillomavirus (HPV) 16, HPV18, and other HR-HPV viral load and the histological classification of cervical lesions: Results from a large-scale cross-sectional study.

    Science.gov (United States)

    Wu, Zeni; Qin, Yu; Yu, Lulu; Lin, Chunqing; Wang, Hong; Cui, Jianfeng; Liu, Bin; Liao, Yiqun; Warren, De'Andre; Zhang, Xun; Chen, Wen

    2017-03-01

    The relationship between HPV viral load and histological grades in the development of cervical cancer is in argument. It is helpful to better understand the association by quantitatively detecting viral load of HPV16, 18, and a pool of 12 other high-risk HPV type (OT) independently on the samples of precancer and cancer. A cross-sectional study was performed in five medical centers of China. Histological diagnosis made by local pathologists was adjudicated via a pathology expert panel. A fully automated real-time PCR test was used for the measurement of HPV16, 18, OT, and human β-globin gene. A total of 2,513 women [1,341 normal, 209 low grade intraepithelial lesion (LSIL), 392 high grade intraepithelial lesion (HSIL), 520 squamous cell carcinoma (SCC), and 51 adenocarcinoma (ADC)] were included. There is a linear increase in the total 14 HPV viral load with histological grade from normal to SCC. This trend was not observed in HPV18 infection but HPV16. The viral load for OT was low in normal, peaked in LSIL and HSIL, and declined in SCC and ADC. In the co-infection of HPV16 and HPV18, HPV16 viral load was significantly higher than HPV18 in LSIL and HSIL. In co-infection of HPV16 and OT, higher HPV16 viral load was also seen in SCC and ADC. Viral load of HPV16 increases with cervical lesion grade and is predominant in cervical cancer. HPV18 viral load is low in precancer, but going up in cancer. OT viral load shows inverse trend of HPV18. J. Med. Virol. 89:535-541, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. Reliability of a rapid hematology stain for sputum cytology

    OpenAIRE

    Gonçalves, Jéssica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Célia Tânia

    2014-01-01

    Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two w...

  13. Hoffman's violet and dahlia as specific stains for animal chromosomes.

    Science.gov (United States)

    Dutt, M K

    1979-03-01

    The paper deals with staining of the chromosomes of animal testicular materials with two basic dyes, Hoffman's violet and dahlia of the triphenylmethane group, following iodine-dye procedure. The important finding, as presented herein, is that iodinated alcohol after staining can be substituted with various acids, both organic as well as inorganic, all of which act as trapping agent preventing leaching of the dye that binds with the chromosomal DNA. It is clear from this study that RNA is not involved by this process of staining, since treatment of stained sections with cold phosphoric acid at 5 degrees C for 20--25 min and then stained also reveals perfect colouration of the chromosomes without any cytoplasmic staining. The in vitro absorption properties of Hoffman's violet have also been presented herein. The probable mechanism of action of these dyes has been suggested.

  14. The evaluation of Lugol's iodine solution staining combined with endoscope for diagnosis of non-erosive reflux disease in clinic.

    Science.gov (United States)

    Zhao, H; Xue, L; Liu, J; Zhang, H-Y; Zhang, D; Song, X-H; He, Y-X; Yang, T; Li, C-Q; Li, Y-Q

    2013-11-01

    To evaluate the value of Lugol's iodine solution staining combined with endoscope on the diagnosis of non-erosive reflux disease (NERD). A total of 96 gastroesophageal reflux disease patients were selected to participate in this study. The patients were stained on esophageal mucosa by Lugol's iodine solution and examined at routine endoscopy. The shallow staining and/or non-staining group patients were treated with esomeprazole and mosapride citrate, and then the changes in Lugol's iodine staining, Gerd Q (Gerd questionnaire) scoring and histological characters of esophageal mucosa were recorded before and after treatment. As the results, a total of 68 patients were diagnosed as NERD, and 36 of 68 patients were observed with uniform staining and 32 of 68 patients were observed with shallow staining and/or non-staining. After 4 weeks for treatment, 28 of 32 patients with shallow staining and/or non-staining became uniform staining and 4 of 32 patients were still with shallow staining and/or non-staining. Before and after treatment, the Gerd Q scoring of uniform staining groups and shallow staining and/or non-staining groups all had a significant difference (p Lugol's iodine solution staining combined with endoscope. Lugol's iodine solution staining combined with routine endoscopy, Gerd Q scoring and histomorphology can be used to evaluate the diagnosis and therapeutic effect of NERD.

  15. Image-based stained glass.

    Science.gov (United States)

    Brooks, Stephen

    2006-01-01

    We present a method of restyling an image so that it approximates the visual appearance of a work of stained glass. To this end, we develop a novel approach which involves image warping, segmentation, querying, and colorization along with texture synthesis. In our method, a given input image is first segmented. Each segment is subsequently transformed to match real segments of stained glass queried from a database of image exemplars. By using real sources of stained glass, our method produces high quality results in this nascent area of nonphotorealistic rendering. The generation of the stained glass requires only modest amounts of user interaction. This interaction is facilitated with a unique region-merging tool.

  16. Histological and immunohistochemical evaluation of sentinel lymph ...

    African Journals Online (AJOL)

    Histological and immunohistochemical evaluation of sentinel lymph nodes in breast cancer at a tertiary hospital in the Western Cape, South Africa. ... To determine effective histological examination of sentinel lymph node (SLN) sections for the detection of metastatic breast carcinoma. Methods. A prospective hospital-based ...

  17. 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.

  18. 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.

  19. 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...

  20. Microdissection of stained archival tissue.

    OpenAIRE

    Gupta, S K; Douglas-Jones, A G; Morgan, J M

    1997-01-01

    In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns...

  1. Application of VRML for three-dimensional interactive real-time comparison of OCT structures with standard histology

    Science.gov (United States)

    Tadrous, Paul J.; Podoleanu, Adrian Gh.; Dobre, George M.; Stamp, Gordon W. H.

    2003-07-01

    Optical coherence tomography (OCT) is a potentially useful 'optical biopsy' tool in medicine. To realize this potential doctors must be able to interpret OCT images with at least the same accuracy as conventional histology. It is therefore important that some accurate method of comparing OCT images with conventional histology be found. Despite numerous OCT vs. conventional histology studies in the literature the methods used for comparison have, by necessity, been approximate because it is not possible to cut a physical tissue section in the same plane as the OCT optical section (partly due to histology processing artefacts). In this paper we present a method of rendering solid tissue volumes with semi-transparency using the Virtual Reality Modelling Language (VRML) and devise a VRML script which allows any two volume data sets to be manipulated within the same region of virtual space. This allows the structure of a whole volume of tissue imaged with OCT to be directly compared with the serial section reconstruction of the conventionally stained histology. As the whole volume is visualized any corresponding tissue regions can be more easily identified despite tissue processing artefacts. We demonstrate the effectiveness of our method usng an ex vivo biopsy of human breast carcinoma.

  2. 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.

  3. 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...... such as retinal vessel, the foveal depression, ciliary arteries, drusen, and nevi. RESULTS: Intra- and interindividual variations in relative choroidal thickness were comparable between the two modalities. After histologic fixation of the three melanoma eyes, the fullness of the choroidal vessels was reduced...... 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...

  4. Direct gram stain and urease test to detect Helicobacter pylori.

    Science.gov (United States)

    Van Horn, K G; Dworkin, B M

    1990-01-01

    Antral biopsies were obtained by gastrointestinal endoscopy on 143 adult patients with dyspeptic symptoms of gastritis or peptic ulcer disease. A direct Gram stain and a direct urease test were performed on each biopsy in addition to culture. Forty-three biopsies (30%) were considered positive for Helicobacter pylori based on culture or histologic examination, or both. Thirty-one biopsies (72% sensitivity) were positive for both direct tests, whereas 95 of 100 negative cultures were negative for both tests. Thirty-eight of the 43 positive biopsies were Gram stain positive (sensitivity, 88%; specificity, 100%). The direct urease test alone was positive at 4 hr for 29 biopsies (sensitivity, 67%; specificity, 100%) and at 24 hr for 38 biopsies (sensitivity, 74%; specificity, 95%). Rapid presumptive diagnosis of H. pylori in antral biopsies was obtained when at least one direct test, Gram stain or urease, was positive.

  5. 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.

  6. 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.

  7. Temporal lobe association fiber tractography as compared to histology and dissection.

    Science.gov (United States)

    Holl, Nathalie; Noblet, Vincent; Rodrigo, Sébastian; Dietemann, Jean L; Mekhbi, Mustapha Ben; Kehrli, Pierre; Wolfram-Gabel, Renée; Braun, Marc; Kremer, Stéphane

    2011-10-01

    To compare the temporal lobe white matter fiber bundles obtained by diffusion tensor imaging-based tractography to that by histology and dissection, and to study the interindividual variability of the obtained tracts. DTI (diffusion tensor imaging) acquisitions (30 directions) were obtained from nine healthy volunteers. Imaging post-processing was performed with FSL (FMRIB Software Library) software. Uncinate fasciculus, longitudinal inferior fasciculus and optic radiations were tracked after positioning of the region of interest (ROI) in predetermined anatomical landmarks. Histological data were obtained by cutting 15 µm coronal sections in one left brain hemisphere and staining with modified Heidenhain-Woelcke myelin stain. Dissection was performed on the left brain hemisphere prepared in accordance with the Klingler method. Tractography of each bundle was compared to histology and dissection data. To highlight the interindividual variability of the considered fiber tracts, all the images were affinely registered on an arbitrarily chosen reference image by considering the B0 images. Fiber tracts were then warped according to the corresponding estimated transformation and an average fiber tract image was then computed. Our results demonstrated a good concordance between tractography of the temporal lobe white matter bundles and dissection and histological data. The interindividual reproducibility of each tract seemed to be good, particularly in the middle part. The variability was more important at both ends, probably in relation to the dispersion of fiber bundles. Diffusion tensor imaging-based tractography of temporal lobe white matter tracts seemed to be in accordance with histological and dissection data. Taking into account some limitations, it could be of particular interest for the presurgical planning of temporal lobectomy.

  8. 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

  9. 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....

  10. Plant Histology: Clearing and the Optical Section.

    Science.gov (United States)

    Freeman, H. E.

    1985-01-01

    Clearing is a simple and rapid technique in which 75 percent lactic acid is used to remove pigments and cytoplasmic contents of fresh leaves, enabling microscopic view of various internal leaf layers. Procedures for using the technique (which helps students gain a more thorough understanding of plant anatomy) are given. (DH)

  11. 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…

  12. Salt stains from evaporating droplets

    NARCIS (Netherlands)

    Shahidzadeh, N.; Schut, M.F.L.; Desarnaud, J.; Prat, M.; Bonn, D.

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls, but also very important in many applications such as purification of pharmaceuticals, deicing of

  13. 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.

  14. 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.

  15. Lugol staining for esophageal carcinoma and influence of radiotherapy on it

    International Nuclear Information System (INIS)

    Miyamoto, Hideo; Adachi, Wataru; Koike, Shoichiro; Koide, Naohiko; Iida, Futoshi

    1993-01-01

    To evaluate the endoscopic staining of esophageal carcinoma with lugol solution, 50 patients who underwent esophagectomy for carcinoma were subjected to this study. Among the 50 patients, 21 received irradiation before surgery. The findings of the lugol staining were compared between endoscopic staining and staining on removed specimens. Non-staining area demonstrated by endoscopic procedure almost agreed with that by the procedure on removed specimen in the non-irradiated group, but both areas of 28.6% cases disagreed in the irradiated group. The extent of non-staining area demonstrated by the procedure of removed specimen was compared with histological extent of carcinoma. The non-staining area on the removed specimen was more extended than histological extent of carcinoma; 10.3% in the non-irradiated group and 71.4% in the irradiated group. As one of the causes of the large non-corresponding rate in the irradiated group, radiation esophagitis was demonstrated. It can be finally concluded that the reliability of endoscopic lugol staining is reduced by preoperative irradiation. (author)

  16. 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.

  17. 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.

  18. 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.

  19. Standardization of the Romanowsky-Giemsa stain: the influence of staining time on the RG-staining pattern.

    Science.gov (United States)

    Schulte, E

    1987-01-01

    In this paper the influence of staining time on the staining pattern of Romanowsky-Giemsa (RG) type stains is investigated. Smears of rabbit bone marrow and of human venous blood were stained with azure B-eosin Y, azure A-eosin Y and with the cationic dyes alone under varying conditions of staining time and dye concentration. The stained smears were investigated by integrating microdensitometry. DNA-polyacrylamide (PAA) model films were stained with azure A-eosin Y, the extinction of the stained model films was determined by spectrophotometry. With increasing staining time the color of the cell nuclei changed from blue to an intense purple, the texture of the nuclear chromatin became more prominent. Prolonged staining resulted in over-staining of the cell nuclei with loss of a distinct chromatin texture. Besides such factors as dye concentration and pH of the staining solution standardization of staining time may be considered necessary for the reproducibility of the RG staining pattern.

  20. 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

  1. 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

  2. Clinical relevance of the utilization of vital Lugol's iodine staining in detection and diagnosis of oral cancer and dysplasia.

    Science.gov (United States)

    Elimairi, Imad; Altay, Mehmet Ali; Abdoun, Omer; Elimairi, Amr; Tozoglu, Sinan; Baur, Dale A; Quereshy, Faisal

    2017-03-01

    The aim of this study is to assess the clinical relevance of the utilization of vital Lugol's iodine staining in detection of oral cancer and dysplastic lesions as well as demarcation of the extent of these lesions. A prospective, cross-sectional, hospital-based study was performed in Khartoum Teaching Dental Hospital, Sudan. Suspicious oral epithelial lesions indicating incisional biopsy were stained with 10 % Lugol's iodine solution and were clinically designated as "negative" for dysplasia or neoplasia when no unstained lesion (USL) area was observed, or as "positive" when a USL area is observed. Incisional biopsies involving both the unstained portion and the stained portion were obtained and histologically evaluated for definitive diagnosis. Forty-five biopsies were obtained from 28 patients. Histopathological examination of the specimens confirmed some degree of dysplasia or neoplasia in all clinically positive specimens with oral squamous cell carcinoma (OSCC) being the most common definitive diagnosis (17 patients, 60.7 %). Clinical relevance level of vital Lugol's iodine staining in detecting oral cancer and dysplasia was found to be 90.9 % (SE = 0.05, P = 0.05). The findings of this study showed that Lugol's iodine is an easy, safe, and effective method of visualizing oral epithelial lesions and differentiating epithelial carcinoma and dysplasia from other benign mucosal lesions. Despite certain limitations, its use is of great value in detection and diagnosis of oral cancer and dysplasia. Lugol's iodine staining can effectively be used in detection of the dysplastic and malignant superficial lesions of the oral epithelium.

  3. 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

  4. 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.

  5. Application of NeuroTrace staining in the fresh frozen brain samples to laser microdissection combined with quantitative RT-PCR analysis.

    Science.gov (United States)

    Benner, Seico; Kakeyama, Masaki; Endo, Toshihiro; Yoshioka, Wataru; Tohyama, Chiharu

    2015-06-20

    The heterogeneity of the brain requires appropriate molecular biological approaches to account for its morphological complexity. Laser-assisted microdissection followed by transcript profiling by quantitative determination has been reported to be an optimal methodology. Nevertheless, not all brain regions can be identified easily without staining, restricting the accuracy and efficiency in sampling. The aim of the present study was to validate whether fixation and staining treatments are suitable for quantitative transcript expression analysis in laser microdissection (LMD) samples. Quantitative RT-PCR was used to determine the absolute transcript expression levels and profiles of samples obtained from the hippocampal dentate gyrus from fresh frozen mice brain sections that had been fixed with ethanol and stained with NeuroTrace. The results were compared with those obtained from unfixed and unstained samples. We found that the quantitative relationship of transcript expression levels between various housekeeping genes and immediate early genes was preserved, although the preparation compromised the yield of the transcripts. In addition, histological and molecular integrities of the fixed and stained specimens were preserved for at least a week at room temperature. Based on the lobe specific profiles of transcripts in the anterior and posterior lobes of the pituitary, we confirmed that no cross-contamination on transcription expressions occurred as a result of the fixation and staining. We have provided detailed information of the procedures on ethanol fixation followed by NeuroTrace staining on the absolute quantitative RT-PCR analysis using microdissected fresh frozen mouse brain tissues. The present study demonstrated that quantitative transcript expression analysis can be conducted reliably on stained tissues. This method is suitable for applications in basic and clinical studies on particular transcript expressions in various regions of the brain.

  6. The effect of decalcifying solutions on hemosiderin staining.

    Science.gov (United States)

    Byard, Roger W; Bellis, Maria

    2010-09-01

    To determine whether routine decalcification may reduce the amount of stainable iron that is visible on tissue sections, samples of liver and lung tissue with excessive iron stores were placed in three standard decalcifying solutions (i) formic acid [33%], formaldehyde [4%], and NaCl [0.85%]; (ii) formic acid [30%], formaldehyde [4%], and water; and (iii) nitric acid [5%] for 24, 48, 72, and 96 h. After exposure to the decalcifying solutions, the tissues were stained with Perls stain. The slides were examined blind and the intensity of iron staining was scored semiquantitatively from 0 to 3+. The trend in all samples over the course of the experiment (96 h) was for reduction in the intensity of hemosiderin staining. As the amount of stainable hemosiderin in tissues may be significantly altered by decalcification, the absence of hemosiderin in tissues adjacent to a fracture site does not necessarily indicate that the injury was acute. © 2010 American Academy of Forensic Sciences.

  7. Novel Non-Toxic Xylene Substitute (SBO) for Histology | Kunhua ...

    African Journals Online (AJOL)

    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.

  8. 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.

  9. Fetal Outcome in Meconium Stained Deliveries

    Science.gov (United States)

    Mundhra, Rajlaxmi; Agarwal, Manika

    2013-01-01

    Objective: To evaluate the foetal outcome in Meconium Stained Amniotic Fluid (MSAF). Material and Methods: This prospective observational study was carried out in the Department of Obstetrics and Gynaecology, North Eastern Indira Gandhi Regional Institute of Health And Medical Sciences, Shillong, India, over a period of eighteen months, from January 2010 to June 2011. A total of 355 pregnant women who had completed more than 37 weeks of gestation, with singleton pregnancies and cephalic presentations were included in this study. One hundred and sixty five cases with MSAF, were thus selected and they were compared with 190 randomly selected controls. Results: Among 165 cases, 27.88 % of the cases had regular visits to the Institute at least 3 times previously, 72.12% cases had no previous visit at all. Primigravidas accounted for a majority of cases and approximately 50% cases had gestational ages of more than 40 weeks Pregnancies complicated with pregnancy induced hypertension had statistically significant higher rates of meconium staining among cases (16.97%), as compared to those among controls (7.89%). 21.81% cases had foetal heart rate abnormalities, as were detected by electronic foetal monitoring and presence of foetal bradycardia was statistically higher in cases compared to that in controls. Casearean section rates were nearly double in cases (49.09%). Neonatal outcome was poor in terms of low Apgar score at birth, birth asphyxia, Meconium Aspiration Syndrome (MAS) and increased neonatal admission among cases as compared to that among controls. Conclusion: Meconium stained amniotic fluid is really worrisome from both, obstetrician’s and paediatrician’s points of view, as it increases the caesarean rates, causes birth asphyxia, MAS and increases neonatal intensive care unit admissions. PMID:24551662

  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. An assessment of the immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol in mammalian tissues

    Energy Technology Data Exchange (ETDEWEB)

    Morley, M.; Gee, D.J.

    1982-10-01

    The use of the indirect immunofluorescence technique as a method for demonstrating the histological localization of tetrahydrocannabinol (delta-THC) has been examined. The experimental protocol was designed in order that optimal staining conditions with respect to temperature, the length of time of incubations and washes, and the dilution of the antisera should be defined. No marked differences were detected between frozen sections of liver from normal and delta-THC-injected mice. Results from radiotracer experiments using human liver suggest that the success of the method is dependent upon the solubility characteristics of the antigen-antibody complex.

  12. 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.

  13. Understanding microwave-stimulated Romanowsky--Giemsa staining of plastic embedded bone marrow.

    Science.gov (United States)

    Horobin, R W; Boon, M E

    1988-01-01

    Bone marrow smears were made and fixed in methanol or formaldehyde. Marrow sections of various thicknesses were also prepared from formaldehyde fixed marrows embedded in paraffin or plastic (glycol methacrylate). The different smears and sections were then stained by a Romanowsky--Giemsa procedure. Some specimens were stained using a standard microwave-stimulated method previously used diagnostically. The effects of technical variations were studied, including degree of microwave irradiation and the staining time. Comparisons of the resulting staining outcomes showed that microwave stimulated Romanowsky--Giemsa staining of plastic sections is a rate controlled process. Unusual aspects of the staining pattern of plastic sections (namely the purple basophilic cytoplasms and nucleoli, and blue chromatin) are due to microwave stimulation and formaldehyde fixation respectively.

  14. Epidermization in the esophageal mucosa: unusual epithelial changes clearly detected by Lugol's staining.

    Science.gov (United States)

    Nakanishi, Y; Ochiai, A; Shimoda, T; Yamaguchi, H; Tachimori, Y; Kato, H; Watanabe, H; Hirohashi, S

    1997-05-01

    A 58-year-old Japanese man with superficial esophageal cancer accompanied by unusual epithelial changes, including esophageal mucosal epidermization, is reported. Staining with Lugol's iodine clearly showed irregular unstained lesions, which could not be seen clearly macroscopically, in the resected specimen. Histologic examination of the irregular unstained areas showed definite granular and horny layers regarded as epidermization, acanthosis with slight nuclear enlargement, and epithelial atrophy. The immunohistochemical staining patterns of keratins in the epidermized and atrophic lesions were similar to those in the epidermis, and the keratin staining patterns of the acanthotic lesion were similar to those of the oral epithelium.

  15. 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.

  16. Dentatorubrothalamic tract localization with postmortem MR diffusion tractography compared to histological 3D reconstruction.

    Science.gov (United States)

    Mollink, J; van Baarsen, K M; Dederen, P J W C; Foxley, S; Miller, K L; Jbabdi, S; Slump, C H; Grotenhuis, J A; Kleinnijenhuis, M; van Cappellen van Walsum, A M

    2016-09-01

    Diffusion-weighted imaging (DWI) tractography is a technique with great potential to characterize the in vivo anatomical position and integrity of white matter tracts. Tractography, however, remains an estimation of white matter tracts, and false-positive and false-negative rates are not available. The goal of the present study was to compare postmortem tractography of the dentatorubrothalamic tract (DRTT) by its 3D histological reconstruction, to estimate the reliability of the tractography algorithm in this specific tract. Recent studies have shown that the cerebellum is involved in cognitive, language and emotional functions besides its role in motor control. However, the exact working mechanism of the cerebellum is still to be elucidated. As the DRTT is the main output tract it is of special interest for the neuroscience and clinical community. A postmortem human brain specimen was scanned on a 7T MRI scanner using a diffusion-weighted steady-state free precession sequence. Tractography was performed with PROBTRACKX. The specimen was subsequently serially sectioned and stained for myelin using a modified Heidenhain-Woelke staining. Image registration permitted the 3D reconstruction of the histological sections and comparison with MRI. The spatial concordance between the two modalities was evaluated using ROC analysis and a similarity index (SI). ROC curves showed a high sensitivity and specificity in general. Highest measures were observed in the superior cerebellar peduncle with an SI of 0.72. Less overlap was found in the decussation of the DRTT at the level of the mesencephalon. The study demonstrates high spatial accuracy of postmortem probabilistic tractography of the DRTT when compared to a 3D histological reconstruction. This gives hopeful prospect for studying structure-function correlations in patients with cerebellar disorders using tractography of the DRTT.

  17. Comparison of immunohistochemical and modified Giemsa stains ...

    African Journals Online (AJOL)

    In 2 cases immunostain could not demonstrate the bacteria but they were identified with modified Giemsa stain while in 5 cases the bacteria were identified by immunostain but not with modified Giemsa stain. The sensitivity of modified Giemsa stain was 85% (CI 66.5-98.8) while the specificity was 89% (CI 60.4 – 97.8).

  18. [Diagnostic stain of helminth eggs (author's transl)].

    Science.gov (United States)

    Cerva, L

    1976-12-01

    A description is given of a diagnostic method for the staining of eggs and larvae of intestinal helminth in smears of both fresh and fixed stool samples. The contents of the eggs and larvae stain red, the background various shades of blue. The most contrasting staining was obtained with thin-walled eggs.

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

    International Nuclear Information System (INIS)

    Petushi, Sokol; Garcia, Fernando U; Haber, Marian M; Katsinis, Constantine; Tozeren, Aydin

    2006-01-01

    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. 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. 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. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. The effect of some fixatives on the staining ability of Sorghum bicolor ...

    African Journals Online (AJOL)

    user

    2006-10-20

    Oct 20, 2006 ... The effect of some fixatives on the staining reactions of the extracts of Sorghum bicolor on tissue sections was studied in order to identify the most appropriate fixative for the stain. Tissue sections taken at postmortem were fixed in 10% formol saline, Carnoy's fluid, Bouin's fluid, Formol sublimate,.

  5. 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.

  6. Histologic classification of gliomas

    NARCIS (Netherlands)

    Perry, Arie; Wesseling, Pieter|info:eu-repo/dai/nl/157872866

    2016-01-01

    Gliomas form a heterogeneous group of tumors of the central nervous system (CNS) and are traditionally classified based on histologic type and malignancy grade. Most gliomas, the diffuse gliomas, show extensive infiltration in the CNS parenchyma. Diffuse gliomas can be further typed as astrocytic,

  7. 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.

  8. 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

  9. 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)

  10. Reliable and durable Golgi staining of brain tissue from human autopsies and experimental animals.

    Science.gov (United States)

    Rosoklija, Gorazd B; Petrushevski, Vladimir M; Stankov, Aleksandar; Dika, Ani; Jakovski, Zlatko; Pavlovski, Goran; Davcheva, Natasha; Lipkin, Richard; Schnieder, Tatiana; Scobie, Kimberley; Duma, Aleksej; Dwork, Andrew J

    2014-06-15

    Golgi stains are notoriously capricious, particularly when applied to human brain. The well-known difficulties, which include complete failure of impregnation, patchy staining, unstable staining, and extensive crystalline deposits in superficial sections, have discouraged many from attempting to use these techniques. A reliable method that produces uniform impregnation in tissue from human autopsies and experimental animals is needed. The method described, "NeoGolgi", modifies previous Golgi-Cox protocols (Glaser and Van der Loos, 1981). Changes include: much longer time (>10 weeks) in Golgi solution, agitation on a slowly rocking platform, more gradual infiltration with Parlodion, more thorough removal of excess staining solution during embedding, and shorter exposure to ammonia after infiltration. The procedure has successfully stained over 220 consecutive frontal or hippocampal blocks from more than 175 consecutive human autopsy cases. Dendritic spines are easily recognized, and background is clear, allowing examination of very thick (200 μm) sections. Stained neurons are evenly distributed within cortical regions. The stain is stable for at least eight years. Most importantly, all stained neurons are apparently well-impregnated, eliminating ambiguity between pathology and poor impregnation that is inherent to other methods. Most methods of Golgi staining are poorly predictable. They often fail completely, staining is patchy, and abnormal morphology is often indistinguishable from poor impregnation. "NeoGolgi" overcomes these problems. Starting with unfixed tissue, it is possible to obtain Golgi staining of predictably high quality in brains from human autopsies and experimental animals. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Morphometric and Histological Analysis of 'Spondylosis Deformans' of Thoracic Region in South-Indian Cadaveric Spines

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    Naveen Kumar

    2017-07-01

    Full Text Available Background: Osteophyte is a bony outgrowth in the vertebral column. Its high prevalence and clinical importance prompted us to conduct this study of thoracic osteophytes. Aim and Objectives: Morphometric and histological study of thoracic osteophytes in the cadaveric vertebral column to understand their development, frequency of occurrence and distribution. Material and Methods: Frequency of occurrence of osteophytes was studied in 50 cadavers of Indian origin over a period of five years. The thoracic part of the vertebral columns were dissected and examined. The vertebral levels of osteophytes, their exact distribution, and morphometric measurements were recorded. A small piece of the osteophyte was removed, processed and stained with Haemetoxylin& Eosin [H & E] stains for histopathological examination. Results: Osteophytes were present in 7 specimens (14%. They were predominantly found on the right side of the lower thoracic vertebral bodies. H & E stained sections of the osteophytes showed features resembling a cancellous bone which strongly indicate that the osteophytes are in development stage, and they develop by the process of endochondral ossification. Conclusion: We found a high incidence of thoracic osteophytes in our study, which mandates further studies in this regard.

  12. Intrapartum amnioinfusion for meconium-stained liquor in developing countries.

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    Moodley, J; Matchaba, P; Payne, A J

    1998-01-01

    Intrapartum amnioinfusion (AI) has been reported to decrease perinatal mortality and morbidity in women with meconium-stained liquor. Such work has not previously been performed at King Edward VIII Hospital (KEH), in a developing country, where the incidence of meconium-stained liquor is said to be extremely high. To establish whether AI during the intrapartum period for meconium-stained liquor decreases Caesarean section rates for fetal distress and decreases perinatal morbidity. Informed consent was obtained from patients in labour who were 3-8 cm dilated, with meconium-staining of the liquor, grades I to III inclusive, and who had a normal cardiotocograph on presentation at term. Sixty patients were included in the trial; 30 had AI. The control group was managed by standard methods. The study group had an amnioinfusion of 0.9% normal saline at 15 ml/min under continuous cardiotocographic monitoring, until a volume of 11 was completed. This was repeated if delivery did not occur within 4 h. The mean pH of umbilical arterial blood was significantly higher in the AI group (7.30 versus 7.23; P = 0.0029). In addition fewer patients in this group developed hypoxic ischaemic encephalopathy (0 versus 2 controls) or meconium aspiration syndrome (1 versus 4 controls). This was not statistically significant. Caesarean section for fetal distress was performed on fewer patients in the AI group (3 versus 7 controls), although this was not statistically significant. These results demonstrate that amnioinfusion is an effective technique for improving the perinatal outcome of pregnancies complicated by meconium-stained liquor in labour. The decrease in Caesarean sections for fetal distress, though not statistically significant in this study, has clinical relevance. Furthermore, this study suggests that amnioinfusion is cost effective in a busy, high-risk labour ward unit and consequently should become standard practice in the management of meconium-stained liquor in labour.

  13. Histological Identification of Propionibacterium acnes in Nonpyogenic Degenerated Intervertebral Discs

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    Ye Yuan

    2017-01-01

    Full Text Available Purpose. Low-virulence anaerobic bacteria, especially the Propionibacterium acnes (P. acnes, have been thought to be a new pathogeny for a series of disc diseases. However, until now, there has been no histological evidence to confirm this link. The purpose of this study was to confirm the presence of P. acnes in nonpyogenic intervertebral discs via histological observation. Method. Degenerated intervertebral discs were harvested from 76 patients with low back pain and/or sciatica but without any symptoms of discitis or spondylodiscitis. The samples were cultured under anaerobic conditions and then examined using 16S rDNA PCR to screen for P. acnes. Samples found to be positive for P. acnes were stained with hematoxylin-eosin (HE and modified Brown-Brenn staining and observed under a microscope. Results. Here, 16 intervertebral discs were found to be positive for P. acnes via 16S rDNA PCR and the prevalence was 21.05% (16/76. Among them, 7 samples had visible microbes stained with HE and modified Brown-Brenn staining. Morphological examination showed the bacteria to be Gram-positive and rod-shaped, so they were considered P. acnes. Conclusion. P. acnes is capable of colonizing some degenerated intervertebral discs without causing discitis, and its presence could be further confirmed by histological evidence. Targeting these bacteria may be a promising therapy method for some disc diseases.

  14. Standardization of Romanowsky stains. The relationship between stain composition and performance.

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    Marshall, P N; Bentley, S A; Lewis, S M

    1978-03-01

    A panel of 17 eminent haematologists has assessed the performance of 5 Romanowsky stains prepared from pure component dyes, comparing the suitability and acceptability of these stains for the preparation of routine blood and bone-marrow films. It was found that the results obtained using the stain described by Marshall et al (1975) were comparable to those obtained using a modification of the stain described by Wittekind et al (1976). The performance of the 3 other stains was less acceptable. Variations in stain formulation have been correlated with stain performance.

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

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    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. Histological and morphometric studies on the dromedary camel epididymis in relation to reproductive activity.

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    Ibrahim, Zarroug Hassan; Singh, Shio Kumar

    2014-10-01

    The reproductive efficiency of male dromedary camels under natural conditions is regarded to be low due to a relatively short breeding season and a long prepubertal period. This study was carried out to investigate the histological and morphometric changes in the Sudanese dromedary camel epididymis in relation to the breeding (rutting) and non-breeding (non-rutting) seasons at monthly intervals up to one year. Specimens were collected from different regions of the epididymis of 48 mature and healthy Sudanese male camels slaughtered at Tambul slaughter house in central Sudan. Paraffin sections of the caput, corpus and cauda regions of the epididymis were stained conventionally for general histology and morphometry. The epididymal stereociliated epithelium showed five types of cells: principal, basal, apical, halo and dark cells and the epithelium was surrounded by a thin layer of lamina propria and a circumtubular smooth muscle coat. No significant seasonal histological or morphometric changes were observed in the caput and corpus epididymides. However, a remarkable decrease in epididymal epithelial thickness and length of stereocilia with consequent increase in luminal diameter was observed in the cauda epididymidis during the rutting season; there was also an increase in the smooth muscle coat thickness. Thus, the results suggest that the epididymis of Sudanese dromedary camel is active throughout the year, but the activity remains low during winter and increases to its maximum during summer, especially during the rainy months. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Histological validation of near-infrared reflectance multispectral imaging technique for caries detection and quantification.

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    Salsone, Silvia; Taylor, Andrew; Gomez, Juliana; Pretty, Iain; Ellwood, Roger; Dickinson, Mark; Lombardo, Giuseppe; Zakian, Christian

    2012-07-01

    Near infrared (NIR) multispectral imaging is a novel noninvasive technique that maps and quantifies dental caries. The technique has the ability to reduce the confounding effect of stain present on teeth. The aim of this study was to develop and validate a quantitative NIR multispectral imaging system for caries detection and assessment against a histological reference standard. The proposed technique is based on spectral imaging at specific wavelengths in the range from 1000 to 1700 nm. A total of 112 extracted teeth (molars and premolars) were used and images of occlusal surfaces at different wavelengths were acquired. Three spectral reflectance images were combined to generate a quantitative lesion map of the tooth. The maximum value of the map at the corresponding histological section was used as the NIR caries score. The NIR caries score significantly correlated with the histological reference standard (Spearman's Coefficient=0.774, p<0.01). Caries detection sensitivities and specificities of 72% and 91% for sound areas, 36% and 79% for lesions on the enamel, and 82% and 69% for lesions in dentin were found. These results suggest that NIR spectral imaging is a novel and promising method for the detection, quantification, and mapping of dental caries.

  18. Histological and morphometric analysis of dilated cardiomyopathy with special reference to collagen IV expression

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    Parul Jain

    2017-01-01

    Full Text Available Introduction: Collagen distribution alterations are well known in dilated cardiomyopathy. There are also changes in microvasculature along with other histomorphorphological features. Aims and Objectives: To study the histomorphological features of DCM along with their quantitative correlation with LVEF. Alterations in collagen IV distribution pattern and microvasculature in DCM were also evaluated. Materials and Methods: The present study includes 34 right ventricular endomyocardial biopsies, 7 explanted native hearts and 41 autopsy control hearts. Sections were taken from lower half of right interventricular septum and stained for H and E, Masson trichrome and immunohistochemistry for CD34, SMA and Collagen IV to study the histological features, pattern of fibrosis, capillary and arteriolar distribution and collagen IV expression respectively. Morphometric analysis was carried out in all cases and controls using Image analysis software Image pro plus 7 and correlated with left ventricular ejection fraction. Results: The histomorphological changes of DCM include myocyte hypertrophy, nucleomegaly, and interstitial fibrosis. Interfiber fibrosis was the commonest. There was evidence of myocarditis, ischemic change and vessel wall alterations. Considerable alteration in Collagen IV distribution was observed with reduction in intensity and proportion of staining around myocytes quantified using Allred scoring against uniform pericellular staining in controls. Morphometric analysis revealed significant increase in nuclear area, myocyte width, percentage of fibrosis and reduction in capillary myocyte ratio in cases as compared to controls. There was no significant difference in arteriolar density. No significant association was observed between morphometric parameters and LVEF. Conclusion: Histomorphological changes in DCM are non-specific. Quantitation of histological parameters cannot be used to predict the disease progression as there was no

  19. Leishman Giemsa cocktail as a new, potentially useful cytological technique comparable to Papanicolaou staining for oral cancer diagnosis.

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    Belgaumi, Ui; Shetty, P

    2013-01-01

    Papanicolaou staining is commonly used for staining exfoliative cytology smears with Romanowsky stains being used sparingly. Leishman Giemsa (LG) cocktail, being a relatively new staining technique, has not been used in exfoliative cytology. This easy, cost-effective and one-step technique warrants further study because of its potential application in screening of oral cancer. To study and evaluate the diagnostic efficiency and reliability of Leishman Giemsa (LG) cocktail in comparison with Papanicolaou (Pap) and May-Grünwald Giemsa (MGG) stains in exfoliated cells for the detection of oral squamous cell carcinoma. Three smears were prepared from each 100 controls (buccal mucosa) and 100 patients, clinically diagnosed with oral squamous cell carcinoma and stained with Pap, MGG and LG cocktail stains. The slides were evaluated for the staining characteristics of nucleus and cytoplasm. The diagnostic efficiency of each stain was evaluated by comparing the cytologic diagnosis of each stain with the histopathological diagnosis. Finally, the diagnostic reliability was evaluated by comparing the three stains with each other and the histologic diagnosis. The data were statistically evaluated with Friedman test, Wilcoxon sign rank test and McNemar chi square test using SPSS15 software. The results from the histologically confirmed cases of squamous cell carcinoma and the number of cases diagnosed by Pap and LG cocktail were almost identical and both were superior to MGG. The P value obtained for the confirmed cases of squamous cell carcinoma in comparison for Pap vs MGG was 0.001, MGG vs LG cocktail was 0.001 and LG cocktail vs Pap was 0.157. Hence, no statistical significant difference was observed between the diagnostic ability of Pap and LG cocktail stains. LG cocktail is an easy, cost-effective and one-step technique comparable to Pap staining; however, it warrants further study in its potential application in screening of oral cancer.

  20. 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

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

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    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

  2. A retrospective study of the clinical, histological, and immunohistochemical manifestations of 5 dogs originally diagnosed histologically as necrotizing scleritis.

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    Denk, Nora; Sandmeyer, Lynne S; Lim, Christine C; Bauer, Bianca S; Grahn, Bruce H

    2012-03-01

    To describe the clinical, histological, and immunohistochemical manifestations of canine necrotizing scleritis. A retrospective examination of the clinical records and samples of ocular tissues from five dogs with a histological diagnosis 'necrotizing scleritis' was completed. Archived, formalin-fixed, paraffin-embedded samples and two control globes were stained with hematoxylin and eosin, Gram, periodic acid-Schiff (PAS) and Masson trichrome stains, and they were immunohistochemically labeled for CD3, CD18, and CD20. Of the five cases reviewed, only two could be confirmed as idiopathic necrotizing scleritis. The other three cases were retrospectively diagnosed as unilateral focal, non-necrotizing scleritis, one as episcleritis and the third was scleritis secondary to a proptosed globe based on our retrospective clinical, histological, and immunohistochemical evaluations. In these two cases, idiopathic necrotizing scleritis manifested as a bilateral, progressive, inflammatory disease of the sclera and cornea that induces significant uveitis. Light microscopic examination confirmed collagen degeneration and granulomatous inflammation. There was no evidence for an infectious etiology based on Gram's and PAS stainings. Immunohistochemical labeling revealed a predominance of B cells in idiopathic, bilateral necrotizing scleritis. Tinctorial staining abnormalities with Masson's trichrome stain were present in scleral collagen of the two cases with idiopathic necrotizing scleritis as well as a case of secondary traumatic scleritis. Based on a limited number of cases, idiopathic canine necrotizing scleritis shares similar histopathological features with non-necrotizing scleritis and episcleritis; however, necrotizing scleritis is B-cell-dominated and bilateral, and significant collagen alterations manifest with Masson's trichrome stain. © 2011 American College of Veterinary Ophthalmologists.

  3. Multicolored stain-free histopathology with coherent Raman imaging.

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    Freudiger, Christian W; Pfannl, Rolf; Orringer, Daniel A; Saar, Brian G; Ji, Minbiao; Zeng, Qing; Ottoboni, Linda; Wei, Ying; Ying, Wei; Waeber, Christian; Sims, John R; De Jager, Philip L; Sagher, Oren; Philbert, Martin A; Xu, Xiaoyin; Kesari, Santosh; Xie, X Sunney; Young, Geoffrey S

    2012-10-01

    Conventional histopathology with hematoxylin & eosin (H&E) has been the gold standard for histopathological diagnosis of a wide range of diseases. However, it is not performed in vivo and requires thin tissue sections obtained after tissue biopsy, which carries risk, particularly in the central nervous system. Here we describe the development of an alternative, multicolored way to visualize tissue in real-time through the use of coherent Raman imaging (CRI), without the use of dyes. CRI relies on intrinsic chemical contrast based on vibrational properties of molecules and intrinsic optical sectioning by nonlinear excitation. We demonstrate that multicolor images originating from CH(2) and CH(3) vibrations of lipids and protein, as well as two-photon absorption of hemoglobin, can be obtained with subcellular resolution from fresh tissue. These stain-free histopathological images show resolutions similar to those obtained by conventional techniques, but do not require tissue fixation, sectioning or staining of the tissue analyzed.

  4. Modified Bismarck brown staining for demonstration of soft tissue mast cells

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    N. Tomov

    2017-09-01

    Full Text Available Bismarck brown staining is a suitable method for demonstration of mast cells in peripheral tissues. However, apart from the intensive color of the mast cell granules, almost no other structures are visible after this staining, which may compromise the results and discourage the investigator. In the present report, we validate the applicability of the Bismarck brown staining of soft tissue and introduce a modification of the method to improve the quality of the histological preparation. Counterstaining with haematoxylin produces specimens with superb contrast and high analytical value. We consider our method involving counterstaining to be superior to the classical Toluidine blue staining, because of the greater contrast of mast cells, which makes evaluation easier, while the preparations are suitable for automated image analysis as well

  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

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    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

  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

    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

  7. Minor salivary gland tumors. A histologic and immunohistochemical study.

    Science.gov (United States)

    Regezi, J A; Lloyd, R V; Zarbo, R J; McClatchey, K D

    1985-01-01

    Clinical, histologic, and immunohistochemical data were reviewed and evaluated on 238 oral minor salivary gland tumors. Benign neoplasms accounted for 65% of the total. Pleomorphic adenomas were the most common of all neoplasms, and mucoepidermoid carcinomas were the most frequently encountered malignancies. Pleomorphic adenomas and adenoid cystic carcinomas stained positive for S-100 protein. Immunohistochemistry was believed to be of potential assistance in diagnosis of salivary gland tumors and in prediction of histogenesis.

  8. Complete androgen insensitivity syndrome: factors influencing gonadal histology including germ cell pathology.

    Science.gov (United States)

    Kaprova-Pleskacova, Jana; Stoop, Hans; Brüggenwirth, Hennie; Cools, Martine; Wolffenbuttel, Katja P; Drop, Stenvert L S; Snajderova, Marta; Lebl, Jan; Oosterhuis, J Wolter; Looijenga, Leendert H J

    2014-05-01

    Patients with complete androgen insensitivity syndrome are at an increased risk for the development of gonadal germ cell cancer. Residual androgen receptor (AR) activity and abnormal gonadal location may influence the survival of atypical germ cells and the development of other histopathological features. To assess this, we evaluated 37 gonads from 19 patients with complete androgen insensitivity (ranging in age from 3 months to 18 years). Histological abnormalities were examined using hematoxylin and eosin-stained sections and sections stained for POU5F1 and KITLG, markers of early changes in germ cells at risk for malignant transformation. Hamartomatous nodules (HNs), Leydig cell hyperplasia (LCH), decreased germ cells, tubular atrophy and stromal fibrosis were more pronounced as age increased (Peffect of inguinal versus abdominal position of the gonads was difficult to assess because inguinal gonads were present primarily in the youngest individuals. In conclusion, many histological changes occur increasingly with age. Expected residual AR activity contributes to better survival of the general germ cell population in (post)pubertal age; however, it did not seem to have an important role in the survival of the germ cells at risk for malignant transformation (defined by POU5F1 positivity and KITLG overexpression) in complete androgen insensitivity. Comparison of the high percentage of patients in our study that were carrying germ cells with delayed maturation or pre-intratubular germ cell neoplasia with previously reported cumulative risk of tumor development in adult patients indicates that not all such precursor lesions in complete androgen insensitivity will progress to invasive germ cell cancer.

  9. Effects of age and diet on rat skin histology.

    Science.gov (United States)

    Thomas, J Regan

    2005-03-01

    To document age-related histologic morphometric changes of rat skin and the effects of calorie restriction on such changes. Fischer 344 rats of three age groups (young, 4 mo; adult, 1 year; old, 24+ months) were procured from ad libitum (AL) diet and calorie-restricted (CR) colonies of the National Institute of Aging and were used for histologic study. Each study group consisted of six animals. Skin samples from the dorsum (DS) and footpad (FP) of these animals were excised and processed for histology with staining techniques for general morphology (hematoxylin-eosin-phloxine) and for differentiation of collagen bundles and elastic fibers (Verhoeff-van Gieson technique). Light microscopic morphometric and stereologic point counting procedures were applied manually to tissue sections to obtain quantitative data on the depth of the epidermis, dermis, and stratum corneum, epidermal nuclear number, and percentage fraction of collagen, elastic fibers, capillaries, and pilosebaceous units. Data were analyzed with two-way of analysis of variance (ANOVA) to determine significant effects of age, diet, and age-diet interaction on these parameters in AL rats and their age-matched cohorts. Significant effects of age, diet, or age-diet interaction were observed in respect of the thickness of epidermis, dermis, stratum corneum of FP, epidermal nuclear number, collagen percentage fraction, and area fraction of capillaries. DS epidermis showed increasing thickness in AL group, but this was reduced in CR rats. A similar trend in DS dermal depth was observed. Fewer capillaries were present in aging CR rats. The DS epidermal nuclear profiles and collagen area fraction also showed effects of diet and age-diet interaction. Aging changes, especially the effect of CR, was more evident in the measured parameters of dorsal skin. No alterations were observed in the distribution of pilosebaceous units and elastic fiber profiles of the skin. The Fischer 344 rat shows many age-related changes

  10. A histological study of peripheral dental caries of equine cheek teeth.

    Science.gov (United States)

    Erridge, Mey E; Cox, Alistair L; Dixon, Padraic M

    2012-01-01

    Peripheral caries of equine cheek teeth is a poorly described disorder and, in particular little is known of its histopathology. Histological examinations of decalcified sections of 21 equine peripheral caries-affected cheek teeth showed two different patterns of cemental lesions; including progressive enlargement of focal, flask-like lesions leading to breakdown of the adjacent cementum, and secondly; a more generalized flaking-off of thin layers of under-run, surface cementum. A thick layer of plaque and food material usually lay on the surface of affected cementum and also within cemental defects. Gram-stained sections showed large numbers of bacteria within the lacunae and canaliculi of affected peripheral cementum and within associated plaque. Pioneer bacteria were also seen within dentinal tubules of adjacent, normal-appearing dentin. Subgingival extension of peripheral caries lesions with localized periodontal destruction was rare. Grading of peripheral caries lesions by gross examination was found to underestimate the severity of the disorder as compared to histological grading.

  11. The nature and frequency of the histological changes found in psoriasis vulgaris.

    Science.gov (United States)

    Beek, C H; van Reede, E C

    1977-01-31

    A total of 272 biopsies (from psoriatic lesions with a diameter of 2-10 cm) were performed on 178 psoriasis patients. Sections from these biopsies were stained with haematoxylin-eosin and by the PAS and trichrome techniques. Each section was investigated for the presence or absence of para- and hyperkeratosis, acanthosis, mitoses, intercellular edema and spongiosis of the epithelial ridges, edema at the top of the connective tissue-papillae, the number of leucocytes penetrating the epidermis at the top of these papillae, classical Munro abscesses, abscesses in the horny layer and Kogoj spongiform pustules, and the results were noted. It was found that the site of the biopsy in the psoriasis lesion, the location of the psoriasis lesion from which material for biopsy was taken and the moment of time during the course of the disease at which the biopsy was performed, had no influence on the histological signs observed in a given patient. In other words, the histological picture of psoriasis plaques from 2-10 cm found in a given patient exhibits a high degree of constancy. Some evidence was found indicating that signs of latent psoriasis may probably be present in the healthy skin of a psoriasis patient. It is generally assumed that the epidermis above the connective tissue-papillae in cases of psoriasis is thinner than normal. Our investigation showed that the reverse is the case: the thickness of the epidermis in the psoriasis efflorescences is significantly greater than in healthy skin nearby in the same patient.

  12. OCT of early dental caries: a comparative study with histology and Raman spectroscopy

    Science.gov (United States)

    Hewko, Mark D.; Choo-Smith, Lin-P'ing; Ko, Alex C.; Leonardi, Lorenzo; Dong, Cecilia C.; Cleghorn, Blaine; Sowa, Michael G.

    2005-03-01

    Early dental caries result from destruction of the tooth's outer mineral matrix by acid-forming bacteria found in dental plaques. Early caries begin as surface disruptions where minerals are leached from the teeth resulting in regions of decreased mineral matrix integrity. Visually, these early carious regions appear as white spots due to the higher backscattering of incident light. With age these areas may become stained by organic compounds. Optical coherence tomography (OCT) examination of human teeth demonstrates a difference in penetration depth of the OCT signal into the carious region in comparison with sound enamel. However, while OCT demonstrates a structural difference in the enamel in the region of the caries, this technique provides little insight into the source of this difference. Raman spectroscopy provides biochemical measures derived from hydroxyapatite within the enamel as well as information on the crystallinity of the enamel matrix. The differences in the biochemical and morphological features of early caries and intact sound enamel are compared. Histological thin sections confirm the observations by OCT morphological imaging while Raman spectroscopy allows for biochemical identification of carious regions by a non-destructive method. Visual examination and conventional radiographic imaging of the intact tooth are used in clinical assessment prior to optical measurements. The combination of OCT, Raman spectroscopy and thin section histology aid in determining the changes that give rise to the visual white spot lesions.

  13. 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

  14. Standard specimens for stain calibration: application to Romanowsky-Giemsa staining.

    Science.gov (United States)

    Turner, J N; Weir, B; Collins, D N

    1990-01-01

    Standardized specimens with reproducible staining properties were fabricated from extracts of biological objects (bovine liver, nucleoprotamine and defatted muscle). The standard specimens were stained with two formulations of the Romanowsky-Giemsa stain (RG), using the same azure B and eosin Y. One formulation used methanol and Sorensen's buffer and the other DMSO and Hepes buffer as solvents. The standard specimens were stained either in the composite stain or in the individual dyes dissolved in the same solvents and at the same concentration as the composite stain. Solution spectroscopy demonstrated different spectra for the two formulations with some wavelength regions varying by more than an order of magnitude. The RG spectra were also very different from those of the individual dyes dissolved at the RG concentration in the respective solvents. The stained standard specimens were analyzed by microspectrophotometry and were found to have spectra similar to those of cell smears. Furthermore, the standard specimens were shown to be a repeatable substrate for stain uptake. The transmitted light intensity from random fields of the same standardized specimen varied +/- 5%. When specimens were stained at the same time, the specimen-to-specimen variation depended on preparation conditions and the measurement wavelength, but was as good as +/- 5% for some conditions. The quantitative stain performance of both formulations was studied and compared. The standardized specimens provide a tool for the quantitative study of staining processes and specimen preparation procedures and for stain calibration.

  15. Histological observation for needle-tissue interactions.

    Science.gov (United States)

    Nakagawa, Yoshiyuki; Koseki, Yoshihiko

    2013-01-01

    We histologically investigated tissue fractures and deformations caused by ex vivo needle insertions. The tissue was formalin-fixed while the needle remained in the tissue. Following removal of the needle, the tissue was microtomed, stained, and observed microscopically. This method enabled observations of cellular and tissular conditions where deformations caused by needle insertions were approximately preserved. For this study, our novel method presents preliminary findings related with tissue fractures and the orientation of needle blade relative to muscle fibers. When the needle blade was perpendicular to the muscle fiber, transfiber fractures and relatively large longitudinal deformations occurred. When the needle blade was parallel to the muscle fiber, interfiber fractures and relatively small longitudinal deformations occurred. This made a significant difference in the resistance force of the needle insertions.

  16. Homogeneously staining regions (HSR) in a human malignant melanoma.

    Science.gov (United States)

    Brieux de Salum, S; Slavutsky, I; Besuschio, S; Pavlovsky, A A

    1984-01-01

    A case of nodular malignant melanoma (level V of Clark's classification) with homogeneously staining regions (HSR) on the long arm of one chromosome #2 is described. Ultrastructural observation of melanosomic and promelanosomic granules near Golgi's vesicles confirmed the histologic diagnosis. Chromosome analysis was performed on nine metaphases from a bone marrow sample and 76 metaphases from culture of the malignant skin tumor. G-banding revealed the presence of a clone with trisomy #8 and another cell line with the HSR marker. This is the first report of HSR in human melanoma cells. As HSR has been found only in malignant cells, we believe that among the many factors that influence the patients' clinical evolution and poor response to treatment, the genic imbalance is of the utmost importance.

  17. Can examination of tissue stained with Oil red O be postponed up to three months?

    Directory of Open Access Journals (Sweden)

    Christoffersen Simone Dorthea

    2014-12-01

    Full Text Available Introduction: As far as we know, there are no known studies on the durability of frozen tissue stained with Oil Red O. The purpose of this study was to examine if the lipid drops in Oil Red O stained tissue keep the original position and color over time (3 months. Further we examined if storage position of the stained tissue makes a difference. Method: We used ten frozen kidney sections stained with Oil Red O. Half of the samples were stored vertically and the other half horizontally, and photos of the same areas were taken within the first 24 hours after staining, and then after 48 hours, 72 hours, 7 days, 14 days, 1 month, 2 months and 3 months respectively. Results and conclusion: No changes in position of the lipids were observed. The color of the staining faded somewhat over time, but it was still possible to distinguish the positive sites from the negative.

  18. 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.

  19. 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.

  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. Purified azure B as a reticulocyte stain.

    Science.gov (United States)

    Marshall, P N; Bentley, S A; Lewis, S M

    1976-01-01

    A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two dyes, varying amounts of an extraneous, particulate dye deposit were present in these preparations, making accuracte counting both tedious and timeconsuming. Purified azure B, on the other hand, gave reproducibly stained, deposit-free preparations. Reticulocyte counts obtained from azure B preparations correlated almost exactly with those determined using new methylene blue. Purified azure B is therefore recommended as a convenient reticulocyte stain for routine use. Images PMID:64475

  2. 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)

  3. Improved staining method for determining the extent of thermal damage to cells.

    Science.gov (United States)

    Sherwood, Margaret E; Flotte, Thomas J

    2007-02-01

    Enzyme histochemical stains of frozen sections have been used by investigators to assess thermal damage. The assessment of thermal damage to cells in lipid-rich tissues such as subcutaneous tissue and sebaceous glands can be difficult due to the quality of frozen sections of such tissues. The purpose of this study is to develop an improved method for this type of evaluation. Thick frozen sections of thermally damaged pig and human skin were stained for lactate dehydrogenase. The sections were fixed in formalin and processed for paraffin-embedded sections. The sections showed well-defined localization of the enzymatic deposits as well as preservation of the tissue architecture. The paraffin-embedded lactate dehydrogenase stained sections provide improved evaluation of thermally damaged tissues, particularly the lipid rich tissues. (c) 2007 Wiley-Liss, Inc.

  4. Stain Removal Assessment of Two Manual Toothbrushes with an Interproximal Tooth Stain Index.

    Science.gov (United States)

    Farrell, Svetlana; Grender, Julie M; Terézhalmy, Geza; Archila, Luis R

    2015-01-01

    To assess a newly developed index to measure interproximal stain and evaluate the stain removal efficacy of two commercially available manual toothbrushes. This was a randomized, examiner-blind, parallel-group, two-treatment clinical trial of two weeks' duration. Subjects qualified for the study if they had an average Modified Lobene Stain Index of ≥ 1.5 from two anterior teeth. At baseline, subjects brushed in front of a mirror for one minute under supervision. All subjects were provided with a standard 0.243% sodium fluoride dentifrice and were randomly assigned either an Oral-B Pulsar manual brush (OBP) or a Colgate Whitening manual brush (CW) to use for two weeks. Stain was reassessed after two weeks of product use. Stain measurements were conducted using the Modified Lobene Stain Index and the new Interproximal Modified Lobene Stain Index, which allows for assessment of stain in hard-to-reach areas using the same area and intensity scales as the Modified Lobene Stain Index. Use of the two manual brushes resulted in statistically significant reductions in surface stain relative to baseline after two weeks of use. Median stain reductions were 78% and 60% for the OBP and CW, respectively, as measured by the Modified Lobene Stain Index. The mean changes in the composite scores from baseline to week two were 1.85 and 1.57 for the two treatment groups, respectively. Statistically significant reductions from baseline were also found for the intensity and extent of stain measures (p brush and 83% reduction with the CW brush. For the gingival sites, the median stain removal percentages were 83% and 50%, respectively For the body region, a median stain removal of 100% was found for both treatment groups. No statistically significant differences were found between the two groups for the mean composite scores for either index. Both manual brushes showed effective stain removal, including interproximal hard-to-reach sites. The Interproximal Modified Lobene Stain Index

  5. Stereomicroscopic examination of stained rectal biopsies in ulcerative colitis and Crohn's disease

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier

    1980-01-01

    Rectal biopsy samples from 22 healthy control subjects, 54 patients with ulcerative colitis, and 34 with Crohn's disease with involvement of the colon or rectum were investigated in a stereomicroscopic study. Samples were stained as whole mounts with Alcian Green before the stereomicroscopic...... the stereomicroscopic findings and the clinical disease activity, the sigmoidoscopic findings, and the histologic activity. Apart from the stereomicroscopic observation of small superficial erosions in one fourth of the biopsies, no changes of diagnostic value were observed in Crohn's disease....

  6. 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

  7. Histological Characterization of Biliary Intraepithelial Neoplasia with respect to Pancreatic Intraepithelial Neoplasia

    Directory of Open Access Journals (Sweden)

    Yasunori Sato

    2014-01-01

    Full Text Available Biliary intraepithelial neoplasia (BilIN is a precursor lesion of hilar/perihilar and extrahepatic cholangiocarcinoma. BilIN represents the process of multistep cholangiocarcinogenesis and is the biliary counterpart of pancreatic intraepithelial neoplasia (PanIN. This study was performed to clarify the histological characteristics of BilIN in relation to PanIN. Using paraffin-embedded tissue sections of surgically resected specimens of cholangiocarcinoma associated with BilIN and pancreatic ductal adenocarcinoma associated with PanIN, immunohistochemical staining was performed using primary antibodies against MUC1, MUC2, MUC5AC, cyclin D1, p21, p53, and S100P. For mucin staining, Alcian blue pH 2.5 was used. Most of the molecules examined here showed similar expression patterns in BilIN and PanIN, in which their expression tended to increase along with the increase in atypia of the epithelial lesions. Significant differences were observed in the increase in mucin production and the expression of S100P in PanIN-1 and the expression of p53 in PanIN-3, when compared with those in BilIN of a corresponding grade. These results suggest that cholangiocarcinoma and pancreatic ductal adenocarcinoma share, at least in part, a common carcinogenic process and further confirm that BilIN can be regarded as the biliary counterpart of PanIN.

  8. 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

  9. 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

  10. The Histological of Ewe Ovarium Post-Intrauterine Transplantation to Pseudopregnancy Rabbit

    OpenAIRE

    Sumarmin, R; Boediono, A; Winarto, A; Yusuf, TL

    2008-01-01

    The objective of this study was to evaluate the histologycal of ewe ovarium post-intrauterin transplantation to pseudopregnancy rabbit. The experiment was concerned with the 1 or 7 days of pseudopregnancy to receive the ewe ovarian transplant. Post transplantation 5, 7 or 9 days of ewe ovarium were recollected. To determine histologically post-intrauterine transplantation of ewe ovarium, the histological preparat was prepared by the paraffin methods followed by HE staining. The result showed ...

  11. On alterations in the refractive index and scattering properties of biological tissue caused by histological processing

    Science.gov (United States)

    Aung, Htet; DeAngelo, Bianca; Soldano, John; Kostyk, Piotr; Rodriguez, Braulio; Xu, M.

    2013-02-01

    Clinical tissue processing such as formalin fixing, paraffin-embedding and histological staining alters significantly the optical properties of the tissue. We document the alterations in the optical properties of prostate cancer tissue specimens in the 500nm to 700nm spectral range caused by histological processing with quantitative differential interference contrast (qDIC) microscopy. A simple model to explain these alterations is presented at the end.

  12. 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.

  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. 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.

  15. 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.

  16. HISTOLOGICAL ANALYSIS AND PERFORMANCE OF SOUR PASSION FRUIT POPULATIONS UNDER DIFFERENT ROOTSTOCKS RESISTANT TO Fusarium spp

    Directory of Open Access Journals (Sweden)

    Moisés Ambrósio

    2018-02-01

    Full Text Available ABSTRACT The present study aimed to describe histological traits of the graft welding process and evaluate the effect of rootstocks on the physical and chemical traits and the productivity of sour passion fruit populations. The experiment was arranged in a randomized block design, in a 4x2 + 4 factorial scheme (cultivars / canopy x rootstocks, three replicates and nine plants per plot. Commercial cultivars and populations and Passiflora alata and P. nitida rootstocks, in addition to ungrafted plants were used as canopy. Cleft grafting was the grafting type adopted. Traits analyzed such as productivity and physical and chemical quality of fruits were submitted to analysis of variance and compared by the Tukey test. The canopy morphology was evaluated according to descriptors of the Ministry of Agriculture, Livestock and Supply. Grafting was carried out for each graft/rootstock combination for histological analysis. Sections were obtained by freehand cuts, stained and mounted on semi-permanent slides, examined under optical microscope and photomicrographed. Rootstocks affected the early cultivation and reduced productivity and number of fruits of passion fruit populations. Grafting did not affect the quality of fruits or the canopy morphology. Considering the complete culture cycle, P. alata species is able to be used as rootstock. The anatomical study demonstrates the occurrence of better compatibility in the connection of P. edulis tissues on P. alata, compared to grafting on P. nitida.

  17. 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.

  18. Effect of sex on histological and histochemical structures of different parts of the kidney in Japanese quail.

    Science.gov (United States)

    Mobini, Behzad; Abdollahi, MohammadHossein

    2016-09-01

    The aim of the present study was to investigate the effect of gender on the histological and histochemical structures of different anatomical regions of the kidney in Japanese quail (Coturnix japonica). Tissue samples from cranial, middle and caudal divisions of each kidney were obtained from 20 male and 20 female adult, healthy Japanese quail. The sections stained with hematoxylin & eosin ( H & E: ), Masson's trichrome, Verhoeff's, Alcian blue (pH 2.5), Periodic acid-Schiff, and Gomori's method for reticulum. Unusual findings of the kidney in Japanese quail were the presence of three types of nephrons, all the connective tissue fibers in capsule and interlobular septa and AB-reactions of the proximal convoluted cells. No significant sex-based differences were found. The various histological structures of the kidney showed no significant differences among different divisions of the left and right kidneys. It is concluded that the general histological and histochemical properties of the kidney in Japanese quail were similar to those of chickens and some other species, but that there were also some differences. One of the major differences was brush border of interdigitating microvilli on luminal surface of collecting ducts in Japanese quail. © 2016 Poultry Science Association Inc.

  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. Potential Value of YAP Staining in Rhabdomyosarcoma.

    Science.gov (United States)

    Ahmed, Atif A; Habeebu, Sultan S; Sherman, Ashley K; Ye, Shui Q; Wood, Nicole; Chastain, Katherine M; Tsokos, Maria G

    2018-03-01

    Rhabdomyosarcoma (RMS) is a common malignancy of soft tissue, subclassified as alveolar (ARMS), pleomorphic (PRMS), spindle cell/sclerosing (SRMS), and embryonal (ERMS) types. The Yes-associated protein (YAP) is a member of the Hippo pathway and a transcriptional regulator that controls cell proliferation. We have studied the immunohistochemical expression of YAP in different RMSs, arranged in tissue microarray (TMA) and whole slide formats. Pertinent clinical data including patient age, gender, tumor location, and clinical stage were collected. Out of 96 TMA cases, 30 cases (31%) were pleomorphic, 27 (28%) were embryonal, 24 (25%) alveolar, and 15 (16%) spindle cell. Positive nuclear YAP staining was seen in the PRMS (17/30, 56.7%), SRMS (7/15, 46.7%), ERMS (19/27 or 70%), and less in ARMS (37.5%). YAP nuclear staining was significantly more prevalent in ERMS than ARMS ( p=0.02). Of the 41 whole slide cases, nuclear staining was detected in all ARMS but was restricted in distribution to 30% staining. These results highlight the role of YAP in RMS tumorigenesis, a fact that can be useful in engineering targeted therapy. Restricted nuclear YAP staining (<30% of cells) may be of value in the diagnosis of ARMS.

  1. M. tuberculosis in Lymph Node Biopsy Paraffin-Embedded Sections

    Directory of Open Access Journals (Sweden)

    Abdurehman Eshete

    2011-01-01

    Full Text Available Background. Tuberculosis lymphadenitis is one of the most common forms of all extrapulmonary tuberculosis. Objective. To evaluate the magnitude of M. tuberculosis from lymph node biopsy paraffin-embedded sections among suspected patients visiting the Jimma University Specialized Hospital. Method. A cross-sectional study design of histological examination among lymph node biopsy paraffin-embedded sections by Ziehl-Neelsen and hematoxylin/eosin staining technique was conducted from December, 2009, to October, 2010, at the Department of Medical Laboratory Science and Pathology. Result. Histopathological examination of the specimens by hematoxylin and eosin staining technique revealed the presence of granulomas. But for the caseation and necrosis they were present in 85% cases of nodal tissue biopsies. From those, 56.7% were from females. The presence of acid-fast bacilli was microscopically confirmed by ZN staining in 37 (61.7% of the nodal tissue biopsies. Conclusion and Recommendation. Tuberculosis lymphadenitis is significantly more common in females. Hence, attention should be given for control and prevention of extrapulmonary tuberculosis.

  2. 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

  3. Utility of Gram stain for the microbiological analysis of burn wound surfaces.

    Science.gov (United States)

    Elsayed, Sameer; Gregson, Daniel B; Lloyd, Tracie; Crichton, Marilyn; Church, Deirdre L

    2003-11-01

    Surface swab cultures have attracted attention as a potential alternative to biopsy histology or quantitative culture methods for microbiological burn wound monitoring. To our knowledge, the utility of adding a Gram-stained slide in this context has not been evaluated previously. To determine the degree of correlation of Gram stain with culture for the microbiological analysis of burn wound surfaces. Prospective laboratory analysis. Urban health region/centralized diagnostic microbiology laboratory. Burn patients hospitalized in any Calgary Health Region burn center from November 2000 to September 2001. Gram stain plus culture of burn wound surface swab specimens obtained during routine dressing changes or based on clinical signs of infection. Degree of correlation (complete, high, partial, none), including weighted kappa statistic (kappa(w)), of Gram stain with culture based on quantitative microscopy and degree of culture growth. A total of 375 specimens from 50 burn patients were evaluated. Of these, 239 were negative by culture and Gram stain, 7 were positive by Gram stain only, 89 were positive by culture only, and 40 were positive by both methods. The degree of complete, high, partial, and no correlation of Gram stain with culture was 70.9% (266/375), 1.1% (4/375), 2.4% (9/375), and 25.6% (96/375), respectively. The degree of correlation for all 375 specimens, as expressed by the weighted kappa statistic, was found to be fair (kappa(w) = 0.32).Conclusion.-The Gram stain is not suitable for the microbiological analysis of burn wound surfaces.

  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. Adversarial Stain Transfer for Histopathology Image Analysis.

    Science.gov (United States)

    Bentaieb, Aicha; Hamarneh, Ghassan

    2018-03-01

    It is generally recognized that color information is central to the automatic and visual analysis of histopathology tissue slides. In practice, pathologists rely on color, which reflects the presence of specific tissue components, to establish a diagnosis. Similarly, automatic histopathology image analysis algorithms rely on color or intensity measures to extract tissue features. With the increasing access to digitized histopathology images, color variation and its implications have become a critical issue. These variations are the result of not only a variety of factors involved in the preparation of tissue slides but also in the digitization process itself. Consequently, different strategies have been proposed to alleviate stain-related tissue inconsistencies in automatic image analysis systems. Such techniques generally rely on collecting color statistics to perform color matching across images. In this work, we propose a different approach for stain normalization that we refer to as stain transfer. We design a discriminative image analysis model equipped with a stain normalization component that transfers stains across datasets. Our model comprises a generative network that learns data set-specific staining properties and image-specific color transformations as well as a task-specific network (e.g., classifier or segmentation network). The model is trained end-to-end using a multi-objective cost function. We evaluate the proposed approach in the context of automatic histopathology image analysis on three data sets and two different analysis tasks: tissue segmentation and classification. The proposed method achieves superior results in terms of accuracy and quality of normalized images compared to various baselines.

  6. Detection Of Concrete Deterioration By Staining

    Science.gov (United States)

    Guthrie, Jr., George D.; Carey, J. William

    1999-09-21

    A method using concentrated aqueous solutions of sodium cobaltinitrite and a rhodamine dye is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR), and to identify degraded concrete which results in a porous or semi-permeable paste due to carbonation or leaching. These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na--K--Ca--Si gels are identified by yellow staining, and alkali-poor, Ca--Si gels are identified by pink staining.

  7. News from the biological stain commission

    DEFF Research Database (Denmark)

    Kiernan, J.A.; Lyon, Hans Oluf

    2008-01-01

    In the three earlier editions of News from the Biological Stain Commission (BSC), under the heading of "Regulatory affairs," the BSC's International Affairs Committee reported on the work of Technical Committee 212, Clinical Laboratory Testing and in Vitro Diagnostic Test Systems of the Internati......In the three earlier editions of News from the Biological Stain Commission (BSC), under the heading of "Regulatory affairs," the BSC's International Affairs Committee reported on the work of Technical Committee 212, Clinical Laboratory Testing and in Vitro Diagnostic Test Systems...

  8. Identification of Cyclospora cayetanensis in stool using different stains.

    Science.gov (United States)

    Negm, A Y

    1998-08-01

    Cyclospora cayetanensis, a newly emerging coccidian protozoa is world-wide in distribution. In the present study, different concentrations and staining techniques were used for identification of Cyclospora. Formol-ether sedimentation and Sheather's sugar flotation were used as concentration techniques and the different stains used were: the modified Ziehl-Neelsen, Giemsa, safranin-methylene blue, modifications of trichrome stain, calcoflour white and finally phenol-auramine. The safranin stain was the best, as it stained all the oocysts of Cyclospora uniformly, besides being rapid and easily applicable in the laboratories. Phenol-auramine stained the oocysts well, where both the wall and internal contents fluoresced brightly. With the calcoflour white stain, only the wall of oocysts took that fluorescent stain. The modified Ziehl-Neelsen stained some of the oocysts well, yet great variability in the staining pattern was noticed. Cyclospora oocysts were not efficiently stained with either trichrome modifications or Giemsa stains.

  9. Histological image segmentation using fast mean shift clustering method.

    Science.gov (United States)

    Wu, Geming; Zhao, Xinyan; Luo, Shuqian; Shi, Hongli

    2015-03-20

    Colour image segmentation is fundamental and critical for quantitative histological image analysis. The complexity of the microstructure and the approach to make histological images results in variable staining and illumination variations. And ultra-high resolution of histological images makes it is hard for image segmentation methods to achieve high-quality segmentation results and low computation cost at the same time. Mean Shift clustering approach is employed for histological image segmentation. Colour histological image is transformed from RGB to CIE L*a*b* colour space, and then a* and b* components are extracted as features. To speed up Mean Shift algorithm, the probability density distribution is estimated in feature space in advance and then the Mean Shift scheme is used to separate the feature space into different regions by finding the density peaks quickly. And an integral scheme is employed to reduce the computation cost of mean shift vector significantly. Finally image pixels are classified into clusters according to which region their features fall into in feature space. Numerical experiments are carried on liver fibrosis histological images. Experimental results demonstrate that Mean Shift clustering achieves more accurate results than k-means but is computational expensive, and the speed of the improved Mean Shift method is comparable to that of k-means while the accuracy of segmentation results is the same as that achieved using standard Mean Shift method. An effective and reliable histological image segmentation approach is proposed in this paper. It employs improved Mean Shift clustering, which is speed up by using probability density distribution estimation and the integral scheme.

  10. 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

  11. Histological evaluation of corneal scar formation in pseudophakic bullous keratopathy.

    Directory of Open Access Journals (Sweden)

    Ting Liu

    Full Text Available PURPOSE: To evaluate histological changes in the corneal stroma in pseudophakic bullous keratopathy. METHODS: Twenty-eight patients (28 eyes with pseudophakic bullous keratopathy underwent therapeutic penetrating keratoplasty at Shandong Eye Institute between January 2006 and November 2011. The patients were divided into two groups according to the duration of bullous keratopathy (1.0 year group, and three buttons from enucleated eyes with choroidal melanoma served as a control. In vivo confocal microscopy examination, hematoxylin-eosin, Masson's trichrome stain and Van Gieson staining were used for microscopic examination. The histological evaluation and scoring of the buttons for morphological changes, including the degree of stromal scars, neovascularization and inflammatory cells within the corneal buttons, were compared. To study the underlying mechanism, connective tissue growth factor (CTGF and TGF-β immunohistochemistry were performed. RESULTS: Confocal microscopy examination and histological evaluation and scoring of the buttons showed that compared with the 1.0 year group (P1.0 year group. CONCLUSIONS: During the progression of pseudophakic bullous keratopathy, stromal scars occurred more often in the patients that had a longer duration of disease. Cytokines such as CTGF and TGF-β1 may play a role in this pathological process and deserve further investigation.

  12. A human visual based binarization technique for histological images

    Science.gov (United States)

    Shreyas, Kamath K. M.; Rajendran, Rahul; Panetta, Karen; Agaian, Sos

    2017-05-01

    In the field of vision-based systems for object detection and classification, thresholding is a key pre-processing step. Thresholding is a well-known technique for image segmentation. Segmentation of medical images, such as Computed Axial Tomography (CAT), Magnetic Resonance Imaging (MRI), X-Ray, Phase Contrast Microscopy, and Histological images, present problems like high variability in terms of the human anatomy and variation in modalities. Recent advances made in computer-aided diagnosis of histological images help facilitate detection and classification of diseases. Since most pathology diagnosis depends on the expertise and ability of the pathologist, there is clearly a need for an automated assessment system. Histological images are stained to a specific color to differentiate each component in the tissue. Segmentation and analysis of such images is problematic, as they present high variability in terms of color and cell clusters. This paper presents an adaptive thresholding technique that aims at segmenting cell structures from Haematoxylin and Eosin stained images. The thresholded result can further be used by pathologists to perform effective diagnosis. The effectiveness of the proposed method is analyzed by visually comparing the results to the state of art thresholding methods such as Otsu, Niblack, Sauvola, Bernsen, and Wolf. Computer simulations demonstrate the efficiency of the proposed method in segmenting critical information.

  13. How Romanowsky stains work and why they remain valuable - including a proposed universal Romanowsky staining mechanism and a rational troubleshooting scheme.

    Science.gov (United States)

    Horobin, R W

    2011-02-01

    An introduction to the nomenclature and concept of "Romanowsky stains" is followed by a brief account of the dyes involved and especially the crucial role of azure B and of the impurity of most commercial dye lots. Technical features of standardized and traditional Romanowsky stains are outlined, e.g., number and ratio of the acidic and basic dyes used, solvent effects, staining times, and fixation effects. The peculiar advantages of Romanowsky staining are noted, namely, the polychromasia achieved in a technically simple manner with the potential for stain intensification of "the color purple." Accounts are provided of a variety of physicochemically relevant topics, namely, acidic and basic dyeing, peculiarities of acidic and basic dye mixtures, consequences of differential staining rates of different cell and tissue components and of different dyes, the chemical significance of "the color purple," the substrate selectivity for purple color formation and its intensification in situ due to a template effect, effects of resin embedding and prior fixation. Based on these physicochemical phenomena, mechanisms for the various Romanowsky staining applications are outlined including for blood, marrow and cytological smears; G-bands of chromosomes; microorganisms and other single-cell entities; and paraffin and resin tissue sections. The common factors involved in these specific mechanisms are pulled together to generate a "universal" generic mechanism for these stains. Certain generic problems of Romanowsky stains are discussed including the instability of solutions of acidic dye-basic dye mixtures, the inherent heterogeneity of polychrome methylene blue, and the resulting problems of standardization. Finally, a rational trouble-shooting scheme is appended.

  14. 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

  15. 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

  16. 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 ...

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

    African Journals Online (AJOL)

    1991-03-16

    Mar 16, 1991 ... methylene blue;8 fluorescence (auramine-phenoI9 and direct immunofluorescence1o. • ll. ); negative staining (periodic acid-. Schiff12. ); and flotation (Sheather's ..... using a direct immunofluorescence assay. Pediatr Infect Dis 1986; 5: 139-142. 12. Horen WP. Detection of Cryptosporidium in human faecal ...

  18. The Language of Stained-Glass Windows

    Science.gov (United States)

    Brew, Charl Anne

    2010-01-01

    The splendor and beauty of stained glass punctuates any room. In this article, the author describes a cross-curriculum project which incorporated the French classes' research and written study of France in the Middle Ages. For the project the author suggested Sainte-Chapelle which is considered a reliquary and was built by Louis IX to house the…

  19. Corneal staining after treatment with topical tetracycline

    NARCIS (Netherlands)

    Lapid-Gortzak, Ruth; Nieuwendaal, Carla P.; Slomovic, Allan R.; Spanjaard, Lodewijk

    2006-01-01

    PURPOSE: The purpose of this paper is to report a case of corneal staining after treatment with topical tetracycline. METHODS: A patient with crystalline keratopathy caused by Streptococcus viridans after corneal transplantation was treated topically with tetracycline eye drops, based on results of

  20. Validation of Romanowsky staining as a novel screening test for the detection of faecal cryptosporidial oocysts.

    Science.gov (United States)

    Brar, A P S; Sood, N K; Singla, L D; Kaur, P; Gupta, K; Sandhu, B S

    2017-03-01

    Cryptosporidiosis is an emerging waterborne protozoan disease and one of the major causes of neonatal diarrhea in humans and animals. But the disease remains under diagnosed due to lack of availability of special stains in majority of laboratories at primary health centers. Therefore, it requires a rapid screening test for routine diagnosis in conventional laboratory set up. In this pursuit, the present study was planned. During this study, fecal samples from 100 representative animals randomly selected from 17 out breaks of bovine calf diarrhea, were stained with modified Ziehl Neelsen staining (mZN) and Leishman's stain to demonstrate cryptosporidial oocysts and for routine fecal examination, respectively. By mZN staining, 25 cases confirmed the presence of cryptosporidial oocysts. However, examination of Leishman's stained fecal smears revealed round hollow unstained bodies resembling cryptosporidia in 20 cases. Therefore, a comparative morphometric analysis was made between the two techniques to determine their relative efficacy in demonstrating cryptosporidia in the feces of affected animals. The analyses showed that the Leishman's stain can be effective in making a presumptive diagnosis of cryptosporidiosis with a little experience. Confirmation of cryptosporidiosis was done by histopathological examination of intestinal sections of calves died during these out breaks. The findings appear to have great clinical value for routine laboratory screening of fecal samples for cryptosporidiosis as conventional Romanowsky stains are readily available and used for multipurpose examination in most of the laboratories at grass root level. Perusal of literature proved this to be the first attempt at easy diagnostics for cryptosporidiosis.

  1. Histopathological and Radiographic Analysis of Dental Follicle of Impacted Teeth Using Modified Gallego’s Stain

    Science.gov (United States)

    Satheesan, Evie; Tamgadge, Avinash; Bhalerao, Sudhir; Periera, Treville

    2016-01-01

    Introduction In the WHO classification of odontogenic tumours, hard tissue formation has been considered as a sub-classification however, this parameter has not been much explored in dental follicle in literature. Epithelial-mesenchymal interactions play an important role in odontogenesis and its associated pathologies; therefore research on dental follicle should also include mesenchymal components along with epithelial components. Additionally, special stains to identify the nature of such depositions in dental follicle have been less explored. Modified Gallego’s stain is such an example which has not been tried in odontogenic lesions which makes this study unique. Aim Aim of this study was to study histopathological variations in dental follicle, the nature of calcification and depositions using Modified Gallego’s stain and to correlate histological features of dental follicle with pericoronal width radiographically. Materials and Methods A prospective histological study of the dental follicles of 50 impacted teeth was carried out to microscopically evaluate the dental follicular tissues for pathological changes, and to correlate it with pericoronal radiolucency. Impacted teeth with pericoronal radiographic width less than 3mm were included in the study and symptomatic teeth were excluded. Further Modified Gallego stain was used to differentiate the nature of hard tissue formation in dental follicle tissues. Results Dental follicle histologically showed pathological changes resembling dentigerous cyst, ameloblastoma, odontogenic fibroma (Simple and WHO Type), clear cell odontogenic tumour, neurofibroma, neurilemmoma and mucoepidermoid carcinoma. Conclusion The dental follicle surrounding an impacted tooth has the potential to differentiate into a wide variety of tissue types, and thus shows the potential for cyst and tumour development which was observed in this study in most of the specimens with normal follicular width radiographically. PMID:27437341

  2. 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.

  3. Clinical and histological findings after intravitreal injection of bevacizumab (Avastin) in a porcine model of choroidal neovascularization

    DEFF Research Database (Denmark)

    Lassota, Nathan; Prause, Jan Ulrik; Scherfig, Erik

    2010-01-01

    PURPOSE: To examine the effect of intravitreally injected bevacizumab (Avastin) on the histological and angiographic morphology of choroidal neovascularization (CNV) in a masked and placebo-controlled animal study. METHODS: Choroidal neovascularization was induced surgically in 11 porcine eyes...... by perforating Bruch's membrane with a retinal perforator. After closure of the ports used for the vitrectomy, which was performed to facilitate the Bruch's membrane rupture, 0.05 ml of either bevacizumab or Ringer-Lactat (placebo) was injected into the vitreous cavity. Eyes were enucleated after 14 days. Fundus...... photographs and fluorescein angiograms (FAs) were obtained immediately prior to enucleation. Sections of formalin- and paraffin-embedded eyes were examined by light microscopy and by immunohistochemical staining. RESULTS: Placebo-injected eyes exhibited the highest propensity to leak, with five of six eyes...

  4. Clinical and histological findings after intravitreal injection of bevacizumsb (Avastin®) in a porcine model of choroidal neovascularisation

    DEFF Research Database (Denmark)

    Lassota, Nathan; Prause, Jan Ulrik; Scherfig, Erik

    2010-01-01

    PURPOSE: To examine the effect of intravitreally injected bevacizumab (Avastin) on the histological and angiographic morphology of choroidal neovascularization (CNV) in a masked and placebo-controlled animal study. METHODS: Choroidal neovascularization was induced surgically in 11 porcine eyes...... by perforating Bruch's membrane with a retinal perforator. After closure of the ports used for the vitrectomy, which was performed to facilitate the Bruch's membrane rupture, 0.05 ml of either bevacizumab or Ringer-Lactat (placebo) was injected into the vitreous cavity. Eyes were enucleated after 14 days. Fundus...... photographs and fluorescein angiograms (FAs) were obtained immediately prior to enucleation. Sections of formalin- and paraffin-embedded eyes were examined by light microscopy and by immunohistochemical staining. RESULTS: Placebo-injected eyes exhibited the highest propensity to leak, with five of six eyes...

  5. 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.

  6. Comparative Assessment of Conventional Papanicolaou and Modified Ultrafast Papanicolaou Stains in Fine Needle Aspiration Samples and Body Fluids.

    Science.gov (United States)

    Arul, P; Eniya, S; Pushparaj, Magesh; Masilamani, Suresh; Kanmani, P; Lingasamy, C

    2018-01-01

    Conventional Papanicolaou (Pap) stain has undergone many modifications; of these, ultrafast Pap stain is the most popular as it shortens the turnaround time of reporting. Application of modified ultrafast Pap (MUFP) stain in the evaluation of fine needle aspiration (FNA) samples and body fluids are scanty. To evaluate the utility of MUFP stain in various FNA samples and body fluids and compare the findings with those of conventional Pap stain. In this cross-sectional study, two wet-fixed and two airdried smears from each sample [301 samples (255 FNA samples and 46 body fluids)] were prepared and stained by the conventional Pap and MUFP stains, respectively. Concordant and discordant rate was calculated. Quality index (QI) of MUFP stain was assessed by background, overall staining, cell morphology, and nuclear characteristics. MUFP-stained smears were also categorized into excellent, good, and fair. The concordance rate for MUFP stain was 100%. QI of MUFP stain for breast, thyroid, lymph node, soft tissue, salivary gland, and body fluids was 0.9, 0.93, 0.95, 1, 0.94, and 1, respectively. Excellent quality of stain was noted in 53.2% and good in 24.6% of the cases allowing easy diagnosis. In 22.2% of fair cases, diagnosis was possible with some difficulties. Our study concluded that MUFP stain could be considered as a rapid and reliable diagnostic tool and can be applied on a regular basis in FNA samples and body fluids to offer immediate diagnosis. However, caution should be taken while reporting certain MUFP-stained smears to avoid over/under diagnosis.

  7. 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.

  8. 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.

  9. A More Accurate Approach to Amyloid Detection and Subtyping: Combining in situ Congo Red Staining and Immunohistochemistry.

    Science.gov (United States)

    Menter, Thomas; Bachmann, Matthias; Grieshaber, Susanne; Tzankov, Alexandar

    2017-01-01

    Amyloidosis is the result of various, differently approachable diseases. It is vital to subtype the amyloid deposits in order to establish and finally treat the underlying disease properly. Besides the classical staining with Congo red, further procedures like immunohistochemical staining are needed for classification. Here, we present a more accurate approach using Congo red/immunohistochemical double staining easily applicable in routine diagnostics. Modifications of the Congo red staining technique and the immunohistochemical procedures were needed in order to combine both staining procedures on one slide. The evaluation was done using conventional light and fluorescence microscopy. By shortening the staining time for Congo red to 10 s and by modification regarding endogenous peroxidase blockage, accurate results could be obtained for evaluating the Congo red/immunohistochemistry double staining using a fluorescence microscope. Sections of 2 μm instead of 4 μm thickness were superior for evaluation, since they increased staining specificity. The combination of Congo red and immunohistochemistry as in situ double staining on one slide is a feasible approach in the diagnosis of amyloidosis. It allows focusing on the fluorescent Congo red-positive areas when evaluating immunohistochemistry, thus avoiding signing out false-positive results. Additionally, it increases the signal-to-noise ratio of the immunohistochemically stained sections on conventional microscopy. © 2016 S. Karger AG, Basel.

  10. Histological Studies on Pancreas of Goose (Anser Albifrons

    Directory of Open Access Journals (Sweden)

    Behzad Mobini

    2011-03-01

    Full Text Available Histological and histochemical studies on the pancreas of goose (Anser albifrons were carried out using special staining and light microscope. The pancreas in goose is serous tubuloacinar gland having exocrine and endocrine part. Smooth muscle fibres were absent in capsule of pancreas. Acinar cells have bizonal shape. Intralobular ducts, Interlobular and main excretory ducts were present within parenchyma. The intralobular ducts were lined with a simple cuboidal epithelium reach interlobular ducts lined with low columnar epithelium. The main excretory ducts were lined by simple to stratified columnar epithelium. The glands inside the connective tissue of the ducts and basophilic staining on the apical surface of pancreatic duct system were found from the interlobular ducts to the main excretory ducts. The endocrine part was consisted of various shapes and sizes of alpha and beta islets. Mixed islets were not observed in the goose pancreas. Parasympathetic ganglia were observed in the exocrine pancreas. No significant differences were noted between males and females.

  11. 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

  12. Offset-sparsity decomposition for enhancement of color microscopic image of stained specimen in histopathology: further results

    Science.gov (United States)

    Kopriva, Ivica; Popović Hadžija, Marijana; Hadžija, Mirko; Aralica, Gorana

    2016-03-01

    Recently, novel data-driven offset-sparsity decomposition (OSD) method was proposed by us to increase colorimetric difference between tissue-structures present in the color microscopic image of stained specimen in histopathology. The OSD method performs additive decomposition of vectorized spectral images into image-adapted offset term and sparse term. Thereby, the sparse term represents an enhanced image. The method was tested on images of the histological slides of human liver stained with hematoxylin and eosin, anti-CD34 monoclonal antibody and Sudan III. Herein, we present further results related to increase of colorimetric difference between tissue structures present in the images of human liver specimens with pancreatic carcinoma metastasis stained with Gomori, CK7, CDX2 and LCA, and with colon carcinoma metastasis stained with Gomori, CK20 and PAN CK. Obtained relative increase of colorimetric difference is in the range [19.36%, 103.94%].

  13. Staining procedure for the detection of microcracks: application to ewe bone.

    Science.gov (United States)

    Portero-Muzy, N R; Chavassieux, P M; Arlot, M E; Chapurlat, R D

    2011-10-01

    Microcracks are one of the determinants of the bone strength and their accumulation may contribute to increased fracture risk. They are detected after bulk staining with various dyes, including basic fuschin, calcein and xylenol orange. The duration of staining usually varies across types of bone and species. The ewe is a large animal with a bone remodeling similar to humans, used as an animal model in bone histomorphometry studies. The aim of the present study was to determine the optimal conditions for bulk staining with xylenol orange of ewe bone. Xylenol orange 5mM in 70% ethanol was applied to iliac crest and vertebral biopsies for 2 or 15 days or 1, 2 or 3 months. After embedding, sections of 40, 50 and 80 μm thick were cut with either a precision diamond wire saw or a microtome. The staining was not visible after 2 or 15 days and was heterogeneous after 1 or 2 months. The quality of 40 and 50 μm thick sections was not preserved compared with those of 80 μm. Microcracks were suitably observed on ewe bone after bulk staining with xylenol orange for 3 months, in 80 μm thick sections. We conclude that the staining procedures should differ when examining ewe or human bone. This may be due to differences in bone matrix composition. Copyright © 2011 Elsevier Inc. All rights reserved.

  14. 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

  15. 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.

  16. Diagnosis of Blastocystis hominis by different staining techniques.

    Science.gov (United States)

    Khalifa, A M

    1999-01-01

    One hundred and fifty stool samples were collected from diarrheic patients of different ages, and examined for Blastocystis hominis by direct smears and concentrated by Sheather's sugar flotation. Staining was done by: Giemsa, two modifications of trichrome stain, modified Ziehl-Neelsen, safranin-methylene blue and two-auramine stains. Out of the 150 cases nine were positive for blastocystosis. The best stains were safranin-methylene blue and modified Ziehl-Neelsen stains. They had the advantage of staining cysts and amoeboid forms besides being rapid and easy to perform. The modified trichrome stains identified 8 ie, less specific and were time consuming. The auramine dyes stained the cyst, both the wall and internal body fluoresced brightly. Giemsa stain was not an efficient stain. Scanning and transmission electron microscopy (SEM, TEM) were performed to study the fine ultrastructure.

  17. 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.

  18. 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.

  19. Microspectrophotometric studies of Romanowsky stained blood cells. II. Comparison of the performance of two standardized stains.

    Science.gov (United States)

    Marshall, P N; Galbraith, W; Navarro, E F; Bacus, J W

    1981-11-01

    This paper describes a comparison of the performance of two standardized Romanowsky blood stains, namely those of Marshall et al. and Wittekind et al., both containing azure B and eosin alone. Stain performance is assessed objectively by the use of three complementary techniques, all based on the visible absorbance spectra of stained cellular substrates. The first of these techniques is a simple comparison of the shapes and heights of the absorbance spectra. The second technique uses the CIE Colorimetric System, and thus permits the quantitation of colour in a manner that agrees with human observation. CIE co-ordinates (chromaticity points, luminance) are calculated directly from absorbance spectra. The third technique is that of spectral subtraction, which yields a set of factors which describe the quantities of component dyes which are bound by the object. This technique, unlike the other two, requires a priori knowledge of the dyes used in the stains, and their spectra when bound to cellular substrates. Although the differences between the two methods are subtle, and hard for the subjective observer to define, the objective methods described here do show statistically significant differences. Wittekind's stain produces less intense staining, except for lymphocyte and monocyte cytoplasms. To the human eye, the differential coloration of these two substrates is more pronounced, but the difference between all nuclei and cytoplasm is less marked. The major difference in the uptake of dye components is in the small quantities of eosin dimer that are bound in this technique.

  20. 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.

  1. 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.

  2. Comparison of methylene blue/gentian violet stain to Gram's stain for the rapid diagnosis of gonococcal urethritis in men.

    Science.gov (United States)

    Taylor, Stephanie N; DiCarlo, Richard P; Martin, David H

    2011-11-01

    We compared a simple, one-step staining procedure using a mixture of methylene blue and gentian violet to Gram stain for the detection of gonococcal urethritis. The sensitivity and specificity of both Gram stain and methylene blue/gentian violet stain were 97.3% and 99.6%, respectively. There was a 100% correlation between the 2 methods.

  3. Morphology and evolutionary biology of the dolphin (Delphinus sp.) brain--MR imaging and conventional histology.

    Science.gov (United States)

    Oelschläger, H H A; Haas-Rioth, M; Fung, C; Ridgway, S H; Knauth, M

    2008-01-01

    Whole brains of the common dolphin (Delphinus delphis) were studied using magnetic resonance imaging (MRI) in parallel with conventional histology. One formalin-fixed brain was documented with a Siemens Trio Magnetic Resonance scanner and compared to three other brains which were embedded in celloidin, sectioned in the three main planes and stained for cells and fibers. The brain of the common dolphin is large, with the telencephalic hemispheres dominating the brain stem. The neocortex is voluminous and the cortical grey matter thin but extremely extended and densely convoluted. There is no olfactory ventricular recess due to the lack of an anterior olfactory system (olfactory bulb and peduncle). No occipital lobe of the telencephalic hemisphere and no posterior horn of the lateral ventricle are present. A pineal organ could not be detected. The brain stem is thick and underlies a very large cerebellum. The hippocampus and mammillary body are small and the fornix is thin; in contrast, the amygdaloid complex is large and the cortex of the limbic lobe is extended. The visual system is well developed but exceeded by the robust auditory system; for example, the inferior colliculus is several times larger than the superior colliculus. Other impressive structures in the brainstem are the peculiar elliptic nucleus, inferior olive, and in the cerebellum the huge paraflocculus and the very large posterior interpositus nucleus. There is good correspondence between MR scans and histological sections. Most of the brain characteristics can be interpreted as morphological correlates to the successful expansion of this species in the marine environment, which was characterized by the development of a powerful sonar system for localization, communication, and acousticomotor navigation. (c) 2007 S. Karger AG, Basel.

  4. Biomechanic and histologic analysis of fibroblastic effects of tendon-to-bone healing by transforming growth factor β1 (TGF-β1) in rotator cuff tears.

    Science.gov (United States)

    Zhang, Chong; Liu, Yu-Jie

    2017-12-01

    To evaluate the effect of transforming growth factor β1 (TGF-β1) on tendon-to-bone reconstruction of rotator cuff tears. Seventy-two rat supraspinatus tendons were transected and reconstructed in situ. At 8 and 16 weeks, specimens of three groups; that is control, L-dose (low dose), and H-dose (high dose) were harvested and underwent a biomechanical test to evaluate the maximum load and stiffness values. Histology sections of the tendon-to-bone interface were identified by hematoxylin-eosin or Masson trichrome stain. Collagen type III was observed by picric acid sirius red staining under polarized light. The level of insulin-like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF) was measured by the enzyme-linked immunosorbent assay (ELISA) method. Collagen type III of the H-dose group had a significant difference in histology structure compared with the L-dose group (P<0.05). The maximum load and stiffness decreased significantly in the control group compared with the values of the L-dose and H-dose groups. The stiffness among the three groups differed significantly at the same postoperative time (P<0.05). Interestingly, progressive reestablishment of collagen type III affected tendon-to-bone healing significantly in the later stages. The H-dose was associated with an increased collagen type III morphology stimulated by TGF-β1.

  5. Presentation Pattern of Histology and Illustrations in the Histology Text Books Commonly Used by the Medical Undergraduates of Bangladesh.

    Science.gov (United States)

    Amin, N F; Shamim, K M

    2017-01-01

    The teaching-learning and assessment of cell and histology have gone through remarkable changes in recent times. Histology text books recommended by renowned medical institutions should be good reflectors of these changes and the present trends. Cell and histology is categorically visual in nature. So, their presentation in text books is likely to show rational and meaningful use of illustrations. From July 2007 to June 2008, this descriptive observational study was done to analyze the presentation pattern of illustrations in two histology text books commonly recommended for the medical undergraduates of Bangladesh through: a) estimating the relative weights given to (i.e., proportions of printed area allotted to) illustrations and text; b) determining the frequencies of 12 'form's of illustration-defining each 'form' on the basis of how it has been created and how it looks like and c) determining the frequencies of 3 'categories' of illustration-on the basis of their utility as teaching-learning and assessment tools. Illustrations have been found to be allotted more than half of the printed area in the two books together. Regarding the 'form' of illustrations, 'schematic diagram and three-dimensional diagram' has the highest mean frequency (30.11%) and followed by 'photomicrograph - routine stain - high power' (17.56%). More than 90% of the illustrations could be categorized either as 'practically the best tool for the purpose' (70.21%) or as 'a complementary tool to another tool' (21.05%) for using in teaching-learning and assessment. By incorporating the findings of the present study with the present-day ideas and trends in cell and histology education in the renowned institutions of the world and with the understanding of the science of medical education, guidelines can be formulated for improving the methods of teaching and assessment in cell and histology for the medical undergraduates of Bangladesh.

  6. 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)

  7. 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.

  8. Histopathological periodic acid-schiff stains of nail clippings as a second-line diagnostic tool in onychomycosis.

    Science.gov (United States)

    Mayer, Eliza; Izhak, Ofer Ben; Bergman, Reuven

    2012-05-01

    The diagnosis of onychomycosis, using direct microscopy and fungal cultures, is often negative despite the presence of disease. Periodic acid-Schiff (PAS) staining of nail clippings, using histopathological processing, may be positive in these cases. It is not always clear, however, whether the fungal elements detected by PAS staining are pathogenic fungi or some are saprophytes. We aimed to study the efficacy of histopathological PAS staining of nail clippings as a second-line diagnostic tool in onychomycosis. The study included 100 consecutive cases in which direct microscopy and fungal cultures from suspected onychomycosis were negative on one occasion or more. The obtained nail clippings were processed for routine histology, stained with hematoxylin and eosin and PAS, and examined microscopically. Of the 100 cases, 38 (38%) showed positive fungal elements. As a result, 9 patients had sought and received oral antifungal therapy and all achieved complete clinical cure. The histological examination also revealed parakeratosis and globules of plasma, which were statistically significantly more common in the fungal infected nail samples. This may indicate an ongoing inflammatory process associated with onychomycosis. Neutrophils and bacteria were not statistically and significantly more common in the fungal infected nails. We conclude that as a second-line diagnostic tool, PAS stain of nail clippings increases markedly the diagnostic yield of onychomycosis and, consequently, the outcome of therapy.

  9. 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...

  10. 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.

  11. A comparative analysis of 7.0-Tesla magnetic resonance imaging and histology measurements of knee articular cartilage in a canine posterolateral knee injury model: a preliminary analysis.

    Science.gov (United States)

    Pepin, Scott R; Griffith, Chad J; Wijdicks, Coen A; Goerke, Ute; McNulty, Margaret A; Parker, Josh B; Carlson, Cathy S; Ellermann, Jutta; LaPrade, Robert F

    2009-11-01

    There has recently been increased interest in the use of 7.0-T magnetic resonance imaging for evaluating articular cartilage degeneration and quantifying the progression of osteoarthritis. The purpose of this study was to evaluate articular cartilage cross-sectional area and maximum thickness in the medial compartment of intact and destabilized canine knees using 7.0-T magnetic resonance images and compare these results with those obtained from the corresponding histologic sections. Controlled laboratory study. Five canines had a surgically created unilateral grade III posterolateral knee injury that was followed for 6 months before euthanasia. The opposite, noninjured knee was used as a control. At necropsy, 3-dimensional gradient echo images of the medial tibial plateau of both knees were obtained using a 7.0-T magnetic resonance imaging scanner. Articular cartilage area and maximum thickness in this site were digitally measured on the magnetic resonance images. The proximal tibias were processed for routine histologic analysis with hematoxylin and eosin staining. Articular cartilage area and maximum thickness were measured in histologic sections corresponding to the sites of the magnetic resonance slices. The magnetic resonance imaging results revealed an increase in articular cartilage area and maximum thickness in surgical knees compared with control knees in all specimens; these changes were significant for both parameters (P .1). These results demonstrate that 7.0-T magnetic resonance imaging provides an alternative method to histology to evaluate early osteoarthritic changes in articular cartilage in a canine model by detecting increases in articular cartilage area. The noninvasive nature of 7.0-T magnetic resonance imaging will allow for in vivo monitoring of osteoarthritis progression and intervention in animal models and humans for osteoarthritis.

  12. 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

  13. [Detection and documentation of masked blood stains with infrared technique].

    Science.gov (United States)

    Du Chesne, A; Bajanowski, T; Brinkmann, B

    1993-01-01

    On dark textiles the visualization of blood stains with the naked eye is either difficult or impossible. In experimental stains and in case work stains we have applied an infrared (IR) video camera in combination with a video printer. As an alternative, an IR goggle was used which could also be connected with a video printer. The results obtained on a variety of different stains and stain carriers are encouraging. Stains showing poor contrast usually become more contrasted. Stains which are partly masked can become complete. Masked stains can become visible. The system is not effective in all combinations of stains and carriers. But it solves a great proportion of formerly problematic cases. Documentation of results is quite easy if a videoprinter is used.

  14. Histological skin morphology enhancement base on molecular hyperspectral imaging technology.

    Science.gov (United States)

    Li, Q; Sun, Z; Wang, Y; Liu, H; Guo, F; Zhu, J

    2014-08-01

    Most traditional skin histological analysis methods are based on the light microscopy images, which can only provide limited information and low contrast results for pathology evaluation. Molecular hyperspectral imaging technology can provide both spatial and spectral information of skin sections, which is a new method for histological skin analysis. The molecular hyperspectral imaging system was developed by coupling an acousto-optic tunable filters adapter to microscopy and the molecular hyperspectral images were analyzed by home-written software with image processing algorithms. Then, the histological structures in skin sections were investigated in several locations to evaluate the potential application of the molecular hyperspectral imaging technique to dermatology. Molecular hyperspectral images of skin sections were obtained. Single-band images, false color images, virtual 3D surface view images, and color-coded spectral clustering results were produced to highlight the skin structures for histological evaluation. Unlike traditional histological analysis with light microscopy, the molecular hyperspectral imaging technology can enhance the visualization of skin structures using their spectral signatures and their gray values. This technology has potential for the diagnosis and histopathologic characterization of different kind of skin cells. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. Robust fadeout profile of an evaporation stain

    Science.gov (United States)

    Witten, T. A.

    2009-06-01

    We propose an explanation for the commonly seen fading in the density of a stain remaining after a droplet has dried on a surface. The density decreases as a power p of the distance from the edge. For thin, dilute drops of general shape this power is determined by a flow stagnation point in the distant interior of the drop. The power p depends on the local evaporation rate J(0) at the stagnation point and the liquid depth h(0) there: p = 1 - 2~ (h(0)/\\bar h)(\\bar J/J(0)) , where \\bar h and \\bar J are averages over the drop surface.

  16. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring

    Directory of Open Access Journals (Sweden)

    Rizzardi Anthony E

    2012-06-01

    Full Text Available Abstract Immunohistochemical (IHC assays performed on formalin-fixed paraffin-embedded (FFPE tissue sections traditionally have been semi-quantified by pathologist visual scoring of staining. IHC is useful for validating biomarkers discovered through genomics methods as large clinical repositories of FFPE specimens support the construction of tissue microarrays (TMAs for high throughput studies. Due to the ubiquitous availability of IHC techniques in clinical laboratories, validated IHC biomarkers may be translated readily into clinical use. However, the method of pathologist semi-quantification is costly, inherently subjective, and produces ordinal rather than continuous variable data. Computer-aided analysis of digitized whole slide images may overcome these limitations. Using TMAs representing 215 ovarian serous carcinoma specimens stained for S100A1, we assessed the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. To evaluate computer-aided image classification, IHC staining within pathologist annotated and software-classified areas of carcinoma were compared for each case. Two metrics for IHC staining were used: the percentage of carcinoma with S100A1 staining (%Pos, and the product of the staining intensity (optical density [OD] of staining multiplied by the percentage of carcinoma with S100A1 staining (OD*%Pos. A comparison of the IHC staining data obtained from manual annotations and software-derived annotations showed strong agreement, indicating that software efficiently classifies carcinomatous areas within IHC slide images. Comparisons of IHC intensity data derived using pixel analysis software versus pathologist visual scoring demonstrated high Spearman correlations of 0.88 for %Pos (p  Virtual slides The virtual slide(s for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1649068103671302

  17. A Histological Autopsy Study of the Thyroid gland in Human ...

    African Journals Online (AJOL)

    SITWALA COMPUTERS

    those which contained autolyzed tissues were excluded from this study. The Paraffin blocked thyroid tissue samples were processed according to standard histopathology laboratory protocols at UTH. This included sectioning and staining with hematoxylin and eosin. The slides were examined at light microscopic level.

  18. The histology of dermal glands of mating Breviceps with comments ...

    African Journals Online (AJOL)

    On arrival in Argentine. some pairs were found. 'glued' together. Skin from dorsal and sternal regions of manually separated adhering pairs was first fixed with. 5OJo GJutaraldehyde/S'ColJidine Buffer (PH 7,9) and later with IOJo Osmium Tetroxide (Os04), and embedded in epoxy resin. Sections of 1 j.lm were stained with.

  19. 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.

  20. Romanowsky staining in cytopathology: history, advantages and limitations.

    Science.gov (United States)

    Krafts, K P; Pambuccian, S E

    2011-04-01

    If the entire discipline of diagnostic cytopathology could be distilled into a single theme, it would be the Papanicolaou stain. Yet it was the Romanowsky stain upon which the discipline of cytopathology was founded. Both stains are used today in the cytopathology laboratory, each for a different and complementary purpose. We trace the history of cytopathological stains and discuss the advantages and limitations of Romanowsky-type stains for cytological evaluation. We also provide suggestions for the advantageous use of Romanowsky-type stains in cytopathology.

  1. 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.

  2. 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.

  3. Histological analysis and 3D reconstruction of winter cereal crowns recovering from freezing: a unique response in oat (Avena sativa L.).

    Science.gov (United States)

    Livingston, David P; Henson, Cynthia A; Tuong, Tan D; Wise, Mitchell L; Tallury, Shyamalrau P; Duke, Stanley H

    2013-01-01

    The crown is the below ground portion of the stem of a grass which contains meristematic cells that give rise to new shoots and roots following winter. To better understand mechanisms of survival from freezing, a histological analysis was performed on rye, wheat, barley and oat plants that had been frozen, thawed and allowed to resume growth under controlled conditions. Extensive tissue disruption and abnormal cell structure was noticed in the center of the crown of all 4 species with relatively normal cells on the outside edge of the crown. A unique visual response was found in oat in the shape of a ring of cells that stained red with Safranin. A tetrazolium analysis indicated that tissues immediately inside this ring were dead and those outside were alive. Fluorescence microscopy revealed that the barrier fluoresced with excitation between 405 and 445 nm. Three dimensional reconstruction of a cross sectional series of images indicated that the red staining cells took on a somewhat spherical shape with regions of no staining where roots entered the crown. Characterizing changes in plants recovering from freezing will help determine the genetic basis for mechanisms involved in this important aspect of winter hardiness.

  4. Effect of LED phototherapy of three distinct wavelengths on fibroblasts on wound healing: a histological study in a rodent model.

    Science.gov (United States)

    de Sousa, Ana Paula Cavalcanti; Santos, Jean N; Dos Reis, João A; Ramos, Taís A; de Souza, José; Cangussú, Maria Cristina T; Pinheiro, Antônio L B

    2010-08-01

    The aim of the present investigation was to evaluate histologically fibroblastic proliferation on dorsal cutaneous wounds in a rodent model treated or not with light-emitting diodes (LEDs) of three wavelengths. Fibroblasts secrete substances essential for wound healing. There are few reports of LED phototherapy on fibroblast proliferation, mainly in vivo. Following approval by the Animal Experimentation Committee of the School of Dentistry of the Federal University of Bahia, we obtained 16 young adult male Wistar rats weighing between 200 and 250 g. Under general anesthesia, one excisional wound was created on the dorsum of each animal; they were then randomly distributed into four groups of four animals each: G0, untreated control; G1, red LED (700 +/- 20 nm, 15 mW, 10 J/cm(2)); G2, green LED (530 +/- 20 nm, 8 mW, 10 J/cm(2)); and G3, blue LED (460 +/- 20 nm, 22 mW, 10 J/cm(2)). The irradiation started immediately after surgery and was repeated every other day for 7 days. Animals were killed 8 days after surgery. The specimens were removed, routinely processed to wax, cut, and stained with hematoxylin/eosin (HE). Fibroblasts were scored by measuring the percentage of these cells occupying the area corresponding to wound healing on stained sections. The quantitative results showed that red LED (700 +/- 20 nm) and green LED (530 +/- 20 nm) showed a significant increase in fibroblast numbers (p LED light is effective in increasing fibroblastic proliferation on rodents.

  5. Influence of sample preparation and reliability of automated numerical refocusing in stain-free analysis of dissected tissues with quantitative phase digital holographic microscopy

    Science.gov (United States)

    Kemper, Björn; Lenz, Philipp; Bettenworth, Dominik; Krausewitz, Philipp; Domagk, Dirk; Ketelhut, Steffi

    2015-05-01

    Digital holographic microscopy (DHM) has been demonstrated to be a versatile tool for high resolution non-destructive quantitative phase imaging of surfaces and multi-modal minimally-invasive monitoring of living cell cultures in-vitro. DHM provides quantitative monitoring of physiological processes through functional imaging and structural analysis which, for example, gives new insight into signalling of cellular water permeability and cell morphology changes due to toxins and infections. Also the analysis of dissected tissues quantitative DHM phase contrast prospects application fields by stain-free imaging and the quantification of tissue density changes. We show that DHM allows imaging of different tissue layers with high contrast in unstained tissue sections. As the investigation of fixed samples represents a very important application field in pathology, we also analyzed the influence of the sample preparation. The retrieved data demonstrate that the quality of quantitative DHM phase images of dissected tissues depends strongly on the fixing method and common staining agents. As in DHM the reconstruction is performed numerically, multi-focus imaging is achieved from a single digital hologram. Thus, we evaluated the automated refocussing feature of DHM for application on different types of dissected tissues and revealed that on moderately stained samples highly reproducible holographic autofocussing can be achieved. Finally, it is demonstrated that alterations of the spatial refractive index distribution in murine and human tissue samples represent a reliable absolute parameter that is related of different degrees of inflammation in experimental colitis and Crohn's disease. This paves the way towards the usage of DHM in digital pathology for automated histological examinations and further studies to elucidate the translational potential of quantitative phase microscopy for the clinical management of patients, e.g., with inflammatory bowel disease.

  6. 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...

  7. Visualizing peripheral nerve regeneration by whole mount staining.

    Directory of Open Access Journals (Sweden)

    Xin-peng Dun

    Full Text Available Peripheral nerve trauma triggers a well characterised sequence of events both proximal and distal to the site of injury. Axons distal to the injury degenerate, Schwann cells convert to a repair supportive phenotype and macrophages enter the nerve to clear myelin and axonal debris. Following these events, axons must regrow through the distal part of the nerve, re-innervate and finally are re-myelinated by Schwann cells. For nerve crush injuries (axonotmesis, in which the integrity of the nerve is maintained, repair may be relatively effective whereas for nerve transection (neurotmesis repair will likely be very poor as few axons may be able to cross between the two parts of the severed nerve, across the newly generated nerve bridge, to enter the distal stump and regenerate. Analysing axon growth and the cell-cell interactions that occur following both nerve crush and cut injuries has largely been carried out by staining sections of nerve tissue, but this has the obvious disadvantage that it is not possible to follow the paths of regenerating axons in three dimensions within the nerve trunk or nerve bridge. To try and solve this problem, we describe the development and use of a novel whole mount staining protocol that allows the analysis of axonal regeneration, Schwann cell-axon interaction and re-vascularisation of the repairing nerve following nerve cut and crush injuries.

  8. 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.

  9. Heparin priming of EUS-FNA needles does not adversely affect tissue cytology or immunohistochemical staining.

    Science.gov (United States)

    Diehl, David L; Mok, Shaffer R S; Khara, Harshit S; Johal, Amitpal S; Kirchner, H Lester; Lin, Fan

    2018-03-01

     Endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) or biopsy (FNB) is an indispensable diagnostic tool. Improvements in needling technique have led to increasing tissue yields. Blood clogging of the needle can cause difficulties with specimen handling and stylet passage, which improves when the needle is primed with heparin before use. However, the effect of heparin on cytology, histology or immunochemistry (IHC) of FNA and FNB specimens is unknown. The goal of the study was to evaluate heparin priming on cytologic/histologic appearance, IHC staining, ease of stylet passage, and specimen bloodiness.  This was a retrospective study of patients undergoing EUS-FNA/FNB. Needle sizes were 25 gauge (g), 22 g, and 19 g. Heparin priming of the needle was done and the stylet replaced ("dry heparin") or suction attached without replacing the stylet ("wet heparin"). Smears and cellblocks were examined by pathologists, and IHC staining were done as needed. Specimen bloodiness was compared with matched controls.  Adequate tissue yields were obtained in all samples (37 heparin, 36 no heparin). Heparin priming did not exhibit negative effects on cytologic or histologic interpretation of the specimens, nor IHC. There was no difference in cellblock bloodiness between the heparin primed needle specimens and the non-heparin control group. .  Heparin priming of EUS-FNA or FNB needles does not negatively affect cytologic or histologic interpretation, nor interfere with IHC. In addition, heparin priming does not increase specimen bloodiness. When the "wet suction" technique is used for EUS-FNA, heparin priming can be used instead of saline priming of the EUS needle.

  10. In vivo histological evaluation of fractional ablative microplasma radio frequency technology using a roller tip: an animal study.

    Science.gov (United States)

    Li, Xiaodan; Fang, Lin; Huang, Luping

    2015-12-01

    The aim of the present study is to investigate the histological characteristics associated with microplasma radio frequency (MPRF) technology in an animal study using different treatment parameters. Two white piglets, aged 6 months, received MPRF treatment using a roller tip; the treatment site was located on the dorsal skin. Four groups of parameters were adopted regarding the performance of the treatment at four zones on the dorsum. Immediately, at 7 days and at 1, 3, and 6 months posttreatment, we observed the healing process and obtained specimens from each treatment zone. Hematoxylin and eosin and Masson stainings of histological sections were performed to assess the degree of tissue injury, the heat effect, the healing process, and neocollagenesis. Heat shock protein (HSP) was also detected using immunohistochemistry. The roller tip generated a fractional treatment, which had a general trend involving an increase in depth and width with increasing pulse energy and decreasing sliding speed. During the wound healing process, dermal neocollagenesis was stimulated, remodeled, and matured gradually. The expression of HSP47 and HPS72 was elevated in the dermis surrounding the microlesions after treatment; it peaked at 1 month posttreatment and became diffuse in the dermis. MPRF is a promising fractional skin resurfacing technique. The roller tip can be used with low risk in the entire treatment zone with rapid healing. An appropriate treatment regimen should be chosen to guarantee therapeutic efficacy and safety.

  11. Histological characterization of Sticholecitha serpentis Prudhoe, 1949 (digenea, bieriidae, sticholecithinae, parasite of Bothrops moojeni Hoge, 1966 (serpentes, viperidae

    Directory of Open Access Journals (Sweden)

    TB. Silva

    2005-12-01

    Full Text Available The aim of this study was to evaluate the morphology of the species Sticholecitha serpentis Prudhoe, 1949 by means of histological procedures. Helminths were previously fixed in Railliet & Henry solution, uncompressed and were included in 2-hydroxyethyl-methacrylate. Longitudinal and transverse serial sections with a 4-mum thickness were performed in a microtome (Leica RM 2165, stained with haematoxylin-eosin and then analyzed in a computerized image analysis system (Qwin Lite 2.5, Leica. Structures of systematic value, such as oral sucker, acetabulum, prepharynx, pharynx, esophagus, intestinal caeca, vitelline glands, ovary, uterus, cirrus pouch and testicles were described. Structures that were poorly visible in total preparations were also observed and described, such as efferent ducts, ejaculatory duct, prostate, seminal vesicle, seminal receptacle, Laurer’s channel, Mehlis’ gland, vitelline ducts, metraterm, genital atrium, digestive glands and excretory vesicle. We demonstrated that histological analysis can supply important data regarding the morphological characterization of S. serpentis and will be able to contribute to systematic studies of trematodes.

  12. Cardiovascular Disease and its Association with Histological Changes of the Left stellate Ganglion

    Directory of Open Access Journals (Sweden)

    Adam Wood

    2010-01-01

    Full Text Available Mounting evidence has demonstrated that the autonomic system plays a role in the morbidity and mortality of certain cardiovascular disease states. Ventricular arrhythmias have been associated with the level of sympathetic activation. We attempted to determine if the presence of fibrosis, a marker for previous ischemic events, correlates with an increase in the number of left stellate ganglion nerve cell bodies which is indicative of hypersympathetic stimulation to the myocardial tissue. Left stellate ganglia were removed, sectioned and prepared using hematoxylin and eosin and Masson's trichrome stain. The interventricular septum of the heart corresponding to the stellate ganglion samples were removed, serially sectioned, and stained with hematoxylin and eosin and Masson's trichrome stain. The samples were described using a grading scale to quantify the percentage of fibrosis. Ganglion nerve cell bodies were then individually counted in three separate high-powered fields. A student's T-test was used to statistically evaluate the data. Stellate ganglions were sampled from 32 cadavers. Fibrosis was present within 72% (23/32 of the interventricular septums that were sampled. Nine interventricular septums were found to be free of fibrosis. For those interventricular septums that were positive for the presence of fibrosis, the mean left stellate ganglion nerve cell bodies was 39.8 (Range: 26-51. For those interventricular septums that were negative for the presence of fibrosis, the mean left stellate ganglion nerve cell bodies was 34.3 (Range: 27-46. The difference between the mean nerve cell bodies for interventricular septums with fibrosis and without fibrosis was found to be statistically significant ( P = 0.048. Histological changes in terms of the number of left stellate ganglion nerve cell bodies seem to be dependent upon the presence of fibrosis within the interventricular septum. Considering fibrosis of the interventricular septum is a marker

  13. Laser therapy in plastic surgery: decolorization in port wine stains

    Science.gov (United States)

    Peszynski-Drews, Cezary; Wolf, Leszek

    1996-03-01

    For the first time laserotherapy is described as a method of port wine stain decolorization in plastic surgery. The authors present their 20-year experience in the treatment of port wine stains with the argon laser and dye laser.

  14. 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.

  15. The degradation of Romanowsky-type blood stains in methanol.

    Science.gov (United States)

    Dean, W W; Stastny, M; Lubrano, G J

    1977-01-01

    The oxidative demethylation of Romanowsky-type stains in methanol has been examined quantitatively with respect to its effect upon the staining of blood smears. Spectral changes in bound dye, observed through two color filters, have been measured for the nuclei and cytoplasm of segmented neutrophils and monocytes utilizing the LARC automated differential analyzer. Stain decomposition in methanol results in a large loss in staining intensity with little change in color. The loss in intensity has been correlated with the observed spectral changes in the degraded stain. High-performance liquid chromatographic analysis of degraded stain samples has shown the products of methanolic degradation to be different from those obtained in aqueous polychroming reactions. To maintain a stain of defined thiazine dye composition and thus defined staining properties, refrigeration is recommended.

  16. 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.

  17. Several staining techniques to enhance the visibility of Acanthamoeba cysts.

    Science.gov (United States)

    El-Sayed, Nagwa Mostafa; Hikal, Wafaa Mohamed

    2015-03-01

    Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of

  18. Histological Value of Duodenal Biopsies

    Directory of Open Access Journals (Sweden)

    Limci Gupta

    2005-01-01

    Full Text Available This study was performed to see the value of histopathological diagnosis in management of patients with duodenal biopsies; to look for correlation of histology and serology in suspected cases of coeliac disease; the reasons for taking duodenal biopsies and whether proper adequate histories are provided on the forms sent with request for histopathological view on duodenal biopsies. Here are the observations of the study followed by the discussion.

  19. 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.

  20. Dye purity and dye standardization for biological staining

    DEFF Research Database (Denmark)

    Lyon, H O

    2002-01-01

    for separating, identifying and assaying dye components. In the second part of the review, descriptions are given of the standardized staining method approach using standard staining methods for assessing stains, and practical responses to stain impurity including commercial quality control, third-party quality...... control and standardization of reagents, protocols and documentation. Finally, reference is made to the current state of affairs in the dye field....

  1. Histological and biochemical evaluations of the liver and kidney of ...

    African Journals Online (AJOL)

    sunny t

    2016-03-16

    Mar 16, 2016 ... confirm the remark through histological section because, significantly elevated levels of ALAT suggest the existence of medical problems such as viral hepatitis, diabetes, congestive heart failure, liver damage, bile duct problems, infectious mononucleosis, or myopathy. So. ALAT is commonly used as a way ...

  2. 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.…

  3. 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

  4. Effects of nicotine on bone during orthodontic tooth movement in male rats. Histological and immunohistochemical study.

    Science.gov (United States)

    Bakathir, Manal A; Linjawi, Amal I; Omar, Samia S; Aboqura, Ahad B; Hassan, Ali H

    2016-10-01

    To investigate the effects of nicotine on orthodontic tooth movement and accompanying histological and immunohistochemical changes in rats. An experimental study conducted at King Abdulaziz University, Jeddah, Saudi Arabia between 2013 and 2014. Thirty-two rats randomly divided into 4 groups. Three were experimental, received daily nicotine injections: group A: 0.37 mg/kg, group B: 0.57 mg/kg, and group C: 0.93 mg/kg. The control group  (group D) received a daily injection of 0.5 mL saline. All rats were subjected to 30 g of orthodontic force on the maxillary left first molars and incisors using a nickel-titanium closed-coil spring. The distance between the 2 teeth was assessed before and after 14 days of force application. Histological, immunohistochemical, and histomorphometric assessments were performed on sections from groups C and D.  Groups C (p less than 0.001) and D (p less than 0.001) showed the significantly greatest and least amounts of tooth movement . The results were statistically dose-dependent. Unbalanced resorption-apposition bone remodeling patterns and increased osteoclast cell distribution were observed in the nicotine group with significantly smaller percentages of bone surface areas mesially and distally (p less than 0.05). Immunohistochemical stains showed low alkaline phosphatase activity and intense tartrate-resistant acid phosphatase activity in the nicotine group.  Nicotine accelerated orthodontic tooth movement with unbalanced bone resorption and apposition patterns around the moving teeth.

  5. 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.

  6. Histological comparison of pulpal inflammation in primary teeth with occlusal or proximal caries.

    Science.gov (United States)

    Kassa, Despoina; Day, Peter; High, Alex; Duggal, Monty

    2009-01-01

    A number of clinical and histological studies have investigated caries-related changes in the primary tooth pulp, but the effect of caries site, as a clinical variable, has not been previously considered. This study sought to compare inflammatory changes within the pulp of primary molars according to the location of the caries lesion (occlusal or proximal). Eighty-three primary molars were extracted under general anaesthesia for caries and/or orthodontic reasons, and were split immediately after removal and fixed in 10% formalin. Teeth were then decalcified, sectioned, and stained with haematoxylin and eosin for histological examination using light microscopy. Caries depth was measured using a graticle, and the site of the caries lesion noted as occlusal or proximal. Samples were further classified into one of five subgroups according to the observed degree of pulpal inflammation. Key findings were that where caries depth was less than 50% of the total dentine thickness, there were no significant differences in inflammatory status according to caries site. In contrast, marked inflammatory changes were significantly more likely throughout the coronal pulp of teeth with proximal caries compared to teeth with occlusal caries where caries depth was equal to, or greater than, 50% of the total dentine thickness (P = 0.017, Fisher's exact test). Primary teeth with proximal carious lesions extending more than 50% through the dentine thickness appear to have more extensive inflammatory pulpal changes than teeth with occlusal caries of a similar depth. This finding has clinical implications and may help inform treatment decisions in the management of primary teeth with deep carious lesions.

  7. Histological study of the biodynamics of iron oxide nanoparticles with different diameters

    Directory of Open Access Journals (Sweden)

    Tsuchiya K

    2011-08-01

    Full Text Available Keiko Tsuchiya1, Norihisa Nitta1, Akinaga Sonoda1, Ayumi Nitta-Seko1, Shinichi Ohta1, Hideji Otani1, Masashi Takahashi1, Kiyoshi Murata1, Katsutoshi Murase2, Satoshi Nohara2, Kenichi Mukaisho31Department of Radiology, Shiga University of Medical Science, Otsu, Shiga, 2The Nagoya Research Laboratory, Meito Sangyo Co, Ltd, Kiyosu, Aichi, 3Department of Pathology, Shiga University of Medical Science, Otsu, Shiga, JapanAbstract: The biodynamics of ultrasmall and small superparamagnetic iron oxide (USPIO and SPIO, respectively particles that were injected intraperitoneally into 36 C57BL/6 mice were investigated chronologically. Their distribution was studied histologically at six time points by measuring iron-positive areas (µm2 in organ sections stained with Prussian blue. The uptake of the differently sized particles was also compared by cultured murine macrophages (J774.1. Iron-positive areas in the liver were significantly larger in the mice injected with USPIO than those injected with SPIO at the first three time points (P < 0.05. The amount of USPIO in the lung parenchyma around the airway was larger than that of SPIO at four time points (P < 0.05; distribution to the lymph nodes was not significantly different. The amount of iron was significantly larger in SPIO- than USPIO-treated cultured cells (P < 0.05. In conclusion, it is suggested that intra peritoneally injected USPIO particles could be used more quickly than SPIO to make Kupffer images of the liver and that both agents could help get lymph node images of similar quality.Keywords: USPIO, SPIO, contrast agent, biodistribution, histology

  8. Value of endoscopic methylene blue and Lugol's iodine double staining and detection of GST-Pi and telomerase in the early diagnosis of esophageal carcinoma.

    Science.gov (United States)

    Zhu, Xuan; Zhang, Shuang-Hong; Zhang, Kun-He; Li, Bi-Ming; Chen, Jiang

    2005-10-21

    To explore the expressions of GST-Pi and telomerase activity in esophageal carcinoma and premalignant lesions and to investigate the value of endoscopic methylene blue (MB) and Lugol's iodine double staining. Seventy-two patients with esophagopathy were sprayed endoscopically with MB and Lugol's iodine in proper order and the areas stained blue and brown, and the area between the blue and brown stains were obtained. Depending on the pattern of mucosal staining, biopsy specimen was obtained. GST-Pi and telomerase activity in specimens were examined by immunohistochemistry and PCR-based silver staining telomeric repeat amplification protocol, respectively. After MB and Lugol's iodine staining, the area between both the colors was obtained in 64 of the 72 patients and the areas were stained blue and brown in all of the 72 patients. Association test of two simultaneous ordinal categorical data showed a correlation between the esophageal mucosal staining and the esophageal histology (PLugol's iodine double staining and detection of GST-Pi and telomerase activity may contribute to the early diagnosis of esophageal carcinoma.

  9. Value of endoscopic methylene blue and Lugol's iodine double staining and detection of GST-Π and telomerase in the early diagnosis of esophageal carcinoma

    Science.gov (United States)

    Zhu, Xuan; Zhang, Shuang-Hong; Zhang, Kun-He; Li, Bi-Ming; Chen, Jiang

    2005-01-01

    AIM: To explore the expressions of GST-Π and telomerase activity in esophageal carcinoma and premalignant lesions and to investigate the value of endoscopic methylene blue (MB) and Lugol's iodine double staining. METHODS: Seventy-two patients with esophagopathy were sprayed endoscopically with MB and Lugol's iodine in proper order and the areas stained blue and brown, and the area between the blue and brown stains were obtained. Depending on the pattern of mucosal staining, biopsy specimen was obtained. GST-Π and telomerase activity in specimens were examined by immunohis-tochemistry and PCR-based silver staining telomeric repeat amplification protocol, respectively. RESULTS: After MB and Lugol's iodine staining, the area between both the colors was obtained in 64 of the 72 patients and the areas were stained blue and brown in all of the 72 patients. Association test of two simultaneous ordinal categorical data showed a correlation between the esophageal mucosal staining and the esophageal histology (PLugol's iodine double staining and detection of GST-Π and telomerase activity may contribute to the early diagnosis of esophageal carcinoma. PMID:16273632

  10. Comparison between Giemsa and Van Geison stains in ...

    African Journals Online (AJOL)

    Trichrome stain (such as Van Geison) is usually used in histopathology laboratory for demonstration of collagenic fibers. Lack of selectivity and tendency of stain to fade makes van Gieson not ideal for collagen demonstration. This study was aimed to compare between Giemsa's and van Gieson's stains in collagen fibers ...

  11. Modified ultrafast Papanicolaou staining technique: A comparative study

    Science.gov (United States)

    Thakur, Moni; Guttikonda, Venkateswara Rao

    2017-01-01

    Introduction: Ultrafast Papanicolaou stain (UFP) was introduced as a hybrid of Romanowsky and Papanicolaou (PAP) stain. It enhances the quality and reduces the time. In the present study, a modified staining technique was adapted where Gill's Hematoxylin was replaced by Harris Hematoxylin. Aims: The aim of the study was to assess the use of the modified ultrafast Papanicolaou (MUFP) stain for fine needle aspiration cytology (FNAC) of head and neck swellings in comparison with the routine PAP stain, hematoxylin and eosin (H and E), and Giemsa. Materials and Methods: Forty FNACs of head and neck swellings were collected. FNAC procedure was performed by standard method; two smears were fixed in 95% propanol and stained with PAP and H and E. Two smears were air dried, 1 was stained with Giemsa, and 1 was rehydrated with normal saline, fixed in alcoholic formalin, and stained with MUFP. Four parameters were considered and scored background, cell morphology, nuclear staining, and overall staining pattern). Results: The quality of MUFP smears were better when compared to routine PAP, H and E, and Giemsa, and was statistically significant by Wilcoxon matched pair test. Conclusions: MUFP stain in comparison to routine PAP, H and E, and Giemsa provides an excellent and suitable alterative in cytological staining for the study of various organs. PMID:28701828

  12. Modified ultrafast Papanicolaou staining technique: A comparative study

    Directory of Open Access Journals (Sweden)

    Moni Thakur

    2017-01-01

    Full Text Available Introduction: Ultrafast Papanicolaou stain (UFP was introduced as a hybrid of Romanowsky and Papanicolaou (PAP stain. It enhances the quality and reduces the time. In the present study, a modified staining technique was adapted where Gill's Hematoxylin was replaced by Harris Hematoxylin. Aims: The aim of the study was to assess the use of the modified ultrafast Papanicolaou (MUFP stain for fine needle aspiration cytology (FNAC of head and neck swellings in comparison with the routine PAP stain, hematoxylin and eosin (H and E, and Giemsa. Materials and Methods: Forty FNACs of head and neck swellings were collected. FNAC procedure was performed by standard method; two smears were fixed in 95% propanol and stained with PAP and H and E. Two smears were air dried, 1 was stained with Giemsa, and 1 was rehydrated with normal saline, fixed in alcoholic formalin, and stained with MUFP. Four parameters were considered and scored background, cell morphology, nuclear staining, and overall staining pattern. Results: The quality of MUFP smears were better when compared to routine PAP, H and E, and Giemsa, and was statistically significant by Wilcoxon matched pair test. Conclusions: MUFP stain in comparison to routine PAP, H and E, and Giemsa provides an excellent and suitable alterative in cytological staining for the study of various organs.

  13. Comparison between Giemsa and Van Geison stains in ...

    African Journals Online (AJOL)

    Rukevwe S. Abraka

    2016-09-14

    Sep 14, 2016 ... Trichrome stain (such as Van Geison) is usually used in histopathology laboratory for demonstration of collagenic fibers. Lack of selectivity and tendency of stain to fade makes van Gieson not ideal for collagen demonstration. This study was aimed to compare between Giemsa's and van Gieson's stains.

  14. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  16. Clinical evaluation of Lugol's iodine staining in the treatment of stage I-II squamous cell carcinoma of the tongue.

    Science.gov (United States)

    Umeda, M; Shigeta, T; Takahashi, H; Minamikawa, T; Komatsubara, H; Oguni, A; Shibuya, Y; Komori, T

    2011-06-01

    Oral squamous cell carcinoma (OSCC) is often surrounded by epithelial dysplasia; leaving it unresected can result in local recurrence. Staining with Lugol's iodine solution detects epithelial dysplasia in oral mucosa, but whether it decreases local recurrence after OSCC surgery is unknown. This study investigated local recurrence rates in patients with early tongue cancer who underwent surgery using Lugol's staining. 93 patients with T1-2N0 tongue SCC underwent partial glossectomy using Lugol's staining during surgery. Resection was performed at least 5mm from the margin of the unstained area. Patients were investigated retrospectively for local recurrence status. Postoperative histology revealed negative surgical margins for SCC or epithelial dysplasia in 81 patients, close margins for SCC in 5, positive margins for mild epithelial dysplasia in 6, and a positive margin for SCC in one. Those with a positive or a close margin for SCC underwent additional resection 2-4 weeks after surgery; one was proved histologically to have residual SCC. No patients developed local recurrence, but 2 died of neck metastasis and 2 of distant metastasis. The 5-year disease specific survival rate was 93.8%. Lugol's staining during surgery can reduce local recurrence and improve survival in patients with early tongue SCC. Copyright © 2011 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  17. Evaluation and histological maturation characteristics of fibrous ...

    African Journals Online (AJOL)

    osseous lesions (FOLS) that are generally considered to be separate entities distinguishable by histologic and radiographic features. The histological maturation of these lesions involves an initial fibrous state, an intermediate mixed and a final ...

  18. 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.

  19. 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... necessary. The test plate should be rocked from side to side a few times to mix the antigen and blood...

  20. 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

  1. 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.

  2. Histological assessment of pulpal responses to resin modified glass ionomer cements in human teeth

    Directory of Open Access Journals (Sweden)

    Ali Eskandarizadeh

    2015-01-01

    Full Text Available Background: The biocompatibility of resin-modified glass ionomers (RMGIs as a lining material is still under question. The present study evaluated the response of the pulp-dentin complex following application of resin-modified glass-ionomer cement, calcium hydroxide and conventional glass-ionomer in deep cavities prepared in human teeth. Materials and Methods: In this controlled clinical trial, 30 deep class V buccal cavities (3 mm × 2 mm × 2 mm were prepared in human premolars treatment planned to be extracted for orthodontic reasons and divided into 3 groups. Groups were lined by a RMGI (Vivaglass, conventional glass Ionomer (Ionocid and calcium hydroxide respectively. The cavities were subsequently filled with amalgam. Each group was then divided into two sub-groups according to time intervals 5 and 30 days. The patients were referred to Kerman Dental School and in accordance with orthodontic treatment plan; premolars were extracted and then prepared for histological assessment. The sections were stained with hematoxylin and eosin and periodic acid Schiff techniques. All of the samples were examined using a number of criteria including odontoblastic changes, inflammatory cells response, reactionary dentin formation and presence of microorganisms. The data were analyzed by Kruskal-Wallis and Mann-Whitney tests. P 0.05. Conclusion: Ionocid and Vivaglass resin-modified glass ionomers can be used as lining materials in human teeth.

  3. Histological and morphometric analyses of seasonal testicular variations in the Jungle Crow (Corvus macrorhynchos).

    Science.gov (United States)

    Islam, Muhammad Nazrul; Zhu, Xiao Bo; Zhu, Ziao Bo; Aoyama, Masato; Sugita, Shoei

    2010-09-01

    A histological and morphometric study was conducted to examine the seasonal testicular variations in the Jungle Crow (Corvus macrorhynchos) of the Kanto area, Japan, from January to July. The paired testes mass, diameter and number of germ cells of the seminiferous tubules, and proportion of seminiferous tubule area and interstitium were examined. Hematoxylin and eosin-stained testis sections and ImageJ Software were used. Paired testes weight was found to increase by 55-fold from January to late March-early May, thereafter declining by 18-fold by June-July. Seminiferous tubule diameter increased fivefold from January to late March-early May, followed a fourfold decrease in June-July. The increase in testes weight correlated well with the increase in the diameter of the seminiferous tubule. In January, the seminiferous tubules constituted 56% of the testicular tissue and the interstitium 44%. During late March-early May, there was very little testicular interstitium (7.9%), and the seminiferous tubules were significantly enlarged (P Jungle Crow has a non-breeding season in January, a pre-breeding season during February-mid March, a main breeding season during late March-early May, a transition period during mid May-late May, and a post-breeding season beginning in June.

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

    Directory of Open Access Journals (Sweden)

    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.  

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  7. STEREOLOGICAL ESTIMATION OF TUBULAR LENGTH FROM THIN VERTICAL SECTIONS

    Directory of Open Access Journals (Sweden)

    Helle V Clausen

    2011-05-01

    Full Text Available In this study tubular structures are represented by stem villous arteries of the human placenta. The architecture of the vascular tree in the human placenta makes it appropriate to use vertical histological sections. Describing tubules, estimates of total length and diameter are informative. The aim of the study was to derive a new stereological estimator of the total length of circular tubules observed in thin vertical sections. Design: Dual perfusion fixed human placentas. Systematic, uniformly random sampling of vertical sections. Five-μm-sections were stained by haematoxylin and eosin (H&E and the vertical axis was identified in all sections. A test system with cycloid test lines was used. Since tubular surface area is proportional to length and diameter, S ∝ πdL, surface-weightening is equivalent to diameter×lengthweightening. As each diameter (extra weight is known, one may eliminate the diameter-weightening by computing the harmonic mean diameter, which is thus the correct, length-weighted mean tubular diameter, dL = d hS . Surface area is estimated in the ordinary way from vertical sections, and with unbiased and robust estimates of S and dL respectively, total length may be estimated L = S / πdhs Conclusion: A new stereological estimator of total length of a circular tubular structure observed in thin vertical sections is presented.

  8. 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

  9. Feulgen type staining of mammalian tissues with Dahlia-SO2.

    Science.gov (United States)

    Dutt, M K

    1977-01-01

    The use of the basic dye, Dahlia, which belongs to triphenylmethane group but without a primary amino group in its molecule has been described as useful in the staining of aldehyde groups of acid hydrolysed DNA in tissue sections following the conventional Feulgen procedure. Dahlia-SO2 prepared with sodium hydrosulphite is highly suitable when used at pH 4-0 to 5-0. The absorption characteristics of the stained nuclei indicate on the peak of maximum absorption at 560 nm, whereas, that of the aqueous dye solution is at 590 nm.

  10. Survival of Pseudomonas aeruginosa in modified Romanowsky staining solutions.

    Science.gov (United States)

    Duffield, Richard; Wong, Hui-San; Trott, Darren J; Hill, Peter B

    2015-08-01

    Anecdotal reports suggest that rapid staining solutions can become contaminated with micro-organisms, especially Pseudomonas aeruginosa. To determine whether inoculation of rapid Romanowsky-type stains with P. aeruginosa results in viable bacterial contamination, which could lead to cross-contamination of slides during cytological staining. Pseudomonas aeruginosa was inoculated into clean and organically contaminated staining solutions (fixative, eosin and methylene blue) and positive (broth) and negative (bleach) control solutions. Subsequent viability and survival were detected by measuring colony-forming units per millilitre at various time points up to 2 weeks. Each sample was stained and microscopically examined to determine whether bacteria were visible. No bacteria could be cultured at any time point from the bleach or fixative solution. In clean eosin and methylene blue staining solutions, viable bacteria were recovered for up to 1 h, but by 24 h all bacteria were dead. In staining solutions contaminated with hair and dead skin cells, bacteria survived in methylene blue for up to 1 h, and viable bacteria persisted in the eosin stain for 2 weeks. In solutions containing viable organisms, the bacteria could be observed by microscopic examination; no bacteria were visible when the solutions contained no viable organisms. Pseudomonas aeruginosa can survive in commonly used staining solutions for variable periods of time, but is unable to proliferate. Although theoretically this might complicate cytological interpretation and subsequent diagnosis, the likelihood of this in clinical practice appears remote when the correct staining technique is used. © 2015 ESVD and ACVD.

  11. 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

  12. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring

    Science.gov (United States)

    2012-01-01

    Abstract Immunohistochemical (IHC) assays performed on formalin-fixed paraffin-embedded (FFPE) tissue sections traditionally have been semi-quantified by pathologist visual scoring of staining. IHC is useful for validating biomarkers discovered through genomics methods as large clinical repositories of FFPE specimens support the construction of tissue microarrays (TMAs) for high throughput studies. Due to the ubiquitous availability of IHC techniques in clinical laboratories, validated IHC biomarkers may be translated readily into clinical use. However, the method of pathologist semi-quantification is costly, inherently subjective, and produces ordinal rather than continuous variable data. Computer-aided analysis of digitized whole slide images may overcome these limitations. Using TMAs representing 215 ovarian serous carcinoma specimens stained for S100A1, we assessed the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. To evaluate computer-aided image classification, IHC staining within pathologist annotated and software-classified areas of carcinoma were compared for each case. Two metrics for IHC staining were used: the percentage of carcinoma with S100A1 staining (%Pos), and the product of the staining intensity (optical density [OD] of staining) multiplied by the percentage of carcinoma with S100A1 staining (OD*%Pos). A comparison of the IHC staining data obtained from manual annotations and software-derived annotations showed strong agreement, indicating that software efficiently classifies carcinomatous areas within IHC slide images. Comparisons of IHC intensity data derived using pixel analysis software versus pathologist visual scoring demonstrated high Spearman correlations of 0.88 for %Pos (p < 0.0001) and 0.90 for OD*%Pos (p < 0.0001). This study demonstrated that computer-aided methods to classify image areas of interest (e.g., carcinomatous areas of tissue specimens) and

  13. Harmonization of the intracellular cytokine staining assay.

    Science.gov (United States)

    Welters, Marij J P; Gouttefangeas, Cécile; Ramwadhdoebe, Tamara H; Letsch, Anne; Ottensmeier, Christian H; Britten, Cedrik M; van der Burg, Sjoerd H

    2012-07-01

    Active immunotherapy for cancer is an accepted treatment modality aiming to reinforce the T-cell response to cancer. T-cell reactivity is measured by various assays and used to guide the clinical development of immunotherapeutics. However, data obtained across different institutions may vary substantially making comparative conclusions difficult. The Cancer Immunotherapy Immunoguiding Program organizes proficiency panels to identify key parameters influencing the outcome of commonly used T-cell assays followed by harmonization. Our successes with IFNγ-ELISPOT and peptide HLA multimer analysis have led to the current study on intracellular cytokine staining (ICS). We report the results of three successive panels evaluating this assay. At the beginning, 3 out of 9 participants (33 %) were able to detect >6 out of 8 known virus-specific T-cell responses in peripheral blood of healthy individuals. This increased to 50 % of the laboratories in the second phase. The reported percentages of cytokine-producing T cells by the different laboratories were highly variable with coefficients of variation well over 60 %. Variability could partially be explained by protocol-related differences in background cytokine production leading to sub-optimal signal-to-noise ratios. The large number of protocol variables prohibited identification of prime guidelines to harmonize the assays. In addition, the gating strategy used to identify reactive T cells had a major impact on assay outcome. Subsequent harmonization of the gating strategy considerably reduced the variability within the group of participants. In conclusion, we propose that first basic guidelines should be applied for gating in ICS experiments before harmonizing assay protocol variables.

  14. Zebrafish embryology and cartilage staining protocols for high school students.

    Science.gov (United States)

    Emran, Farida; Brooks, Jacqueline M; Zimmerman, Steven R; Johnson, Susan L; Lue, Robert A

    2009-06-01

    The Life Sciences-Howard Hughes Medical Institute Outreach Program at Harvard University supports high school science education by offering an on-campus program for students and their teachers to participate in investigative, hands-on laboratory sessions. The outreach program has recently designed and launched a successful zebrafish embryology protocol that we present here. The main objectives of this protocol are to introduce students to zebrafish as a model research organism and to provide students with direct experience with current techniques used in embryological research. The content of the lab is designed to generate discussions on embryology, genetics, fertilization, natural selection, and animal adaptation. The protocol produces reliable results in a time-efficient manner using a minimum of reagents. The protocol presented here consists of three sections: observations of live zebrafish larvae at different developmental stages, cartilage staining of zebrafish larvae, and a mutant hunt involving identification of two zebrafish mutants (nacre and chokh). Here, we describe the protocol, show the results obtained for each section, and suggest possible alternatives for different lab settings.

  15. [Histological and histochemical study of hyperplastic and dysplastic lesions of the large intestine in Cebus Apella (primate) treated with 1,2-dimethylhydrazine].

    Science.gov (United States)

    Sánchez Negrette, M; Borda, J T; Montenegro, M A; Lertora, J W

    1998-01-01

    The main objective of this study was to assess the histological changes of colon ephitelium in Cebus apella induced by 1,2-dimethylhydrazine (DMH) administration. Twelve monkeys, males, (aged x: 30 months) with an average body weight of 2,800 g were utilized. The DMH was injected subcutaneously at 25 mg/kg and continued once a week for 16 weeks. The body weight was assessed once a week during the first 4 months and every 30 days until the end of the experience. Histological changes of intestinal ephitelium and mucins were assessed at the end of the experience in specimens sectioned at 5 microns, stained with Haematoxylin and Eosin, PAS and Alcian blue pH 2.5. The histological and histochemical study permitted to characterize the normal morphology, as well as the mucins characteristics in the three regions: caecum, transverse colon and distal colon. The histological changes in the DMH treated animals were hyperplasia, dysplasia and mucins decreasing. The hyperplastic changes were localizated in glandular crypts, and in the epithelio located over the lymphoid nodules. The dysplastic crypts were observed in the transverse colon and in the last portion of distal colon. These lesions were located in the upper portion as well as the bottom of the mucosa. A decrease of neutral and acids mucopolysaccharides were observed in the crypts. The results of this study suggest that the DMH induced hyperplastic changes in the crypts and in the epithelium located over the lymphoid nodules and dysplastic focus, as well as a decrease of neutral and acids mucopolysaccharides.

  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. 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

  18. 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

  19. TREATMENTS TO MINIMIZE EXTRACTIVES STAIN IN WESTERN RED CEDAR

    OpenAIRE

    Rod Stirling,; Paul I. Morris

    2012-01-01

    Under certain conditions involving uneven exposure to weather, stains related to the extractives can reduce the aesthetic appeal of western red cedar in exterior applications such as fence boards, siding, and sidewall shingles. Selected chemical treatments were evaluated for their ability to inhibit the formation of extractives stain. DDACarbonate, alkyl amine oxide, and combinations thereof delayed extractives stain formation in an accelerated field test, with higher loadings having greater ...

  20. 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 (<3 J/cm(2) ) and is accompanied by a drastic reduction in porphyrin fluorescence from the Soret band. 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.

  1. Studies on the blue-staining fungi of pine wood

    Directory of Open Access Journals (Sweden)

    A. Strzelczyk

    2014-11-01

    Full Text Available The aim of this was were to examine associations of bule-staining fungi which occur on pine wood to determine the interactions between fungi and to check the suscebility of these fungi to commonly used fungieides. The stron antagonism of members of the Trichoderma genus against the blue-staining fungi was demonstrated, Members of genera Pullularia, Hormiscium and Hormodendrum were strongy inhibited by stains of Trichoderma Ophiostoma strains were less susceptible to inhibition by this antagonist.

  2. New versatile staining reagents for biological transmission electron microscopy that substitute for uranyl acetate.

    Science.gov (United States)

    Nakakoshi, Masamichi; Nishioka, Hideo; Katayama, Eisaku

    2011-12-01

    Aqueous uranyl acetate has been extensively used as a superb staining reagent for transmission electron microscopy of biological materials. However, recent regulation of nuclear fuel material severely restricts its use even for purely scientific purposes. Since uranyl salts are hazardous due to biological toxicity and remaining radioactivity, development of safe and non-radioactive substitutes is greatly anticipated. We examined two lanthanide salts, samarium triacetate and gadolinium triacetate, and found that 1-10% solution of these reagents was safe but still possess excellent capability for staining thin sections of plastic-embedded materials of animal and plant origin. Although post-fixation with osmium tetroxide was essential for high-contrast staining, post-staining with lead citrate could be eliminated if a slow-scan CCD camera is available for observation. These lanthanide salts can also be utilized as good negative-staining reagents to study supramolecular architecture of biological macromolecules. They were not as effective as a fixative of protein assembly, reflecting the non-hazardous nature of the reagents.

  3. An experimental teaching-learning program in histology.

    Science.gov (United States)

    Li, Enzhong; Chen, Yanfang

    2011-01-01

    This article presents an experimental teaching and learning program used in a histology course with first-year students in the first term of their studies in the Faculty of Veterinary Medicine at Huanghuai University, China. Eighty students were divided randomly into two sections (n=40/section). Students in one section were taught using a hybrid case-based and lecture style (experimental section) and the second section was taught using a traditional teaching style (control section). To assess achievement, examinations were administered at the end of every textbook chapter and a final examination was carried out at the end of the course. To determine students' attitude to the teaching styles used, a questionnaire survey was conducted at the end of the term. Results showed that students preferred the experimental style. In the experimental section, students' scores were significantly higher than those of students in the control section both in the chapter and the final examinations (pstyle is superior to the traditional style of teaching and learning histology.

  4. Black stain and dental caries in Filipino schoolchildren.

    Science.gov (United States)

    Heinrich-Weltzien, Roswitha; Monse, Bella; van Palenstein Helderman, Wim

    2009-04-01

    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. This study was conducted to assess the prevalence of black stain on teeth of Filipino children and to determine a possible association between black stain and caries levels. The study was designed to test the following hypotheses: (i) the prevalence of black stain does not differ between children from schools with oral health intervention programs and those from schools without an intervention program, (ii) the prevalence of black stain does not differ in children attending easily accessible and remote schools, (iii) caries prevalence and caries experience do not differ in children with and without black stain and (iv) the caries distribution at the surface level does not differ in children with and without black stain. In total, 32 elementary schools were included. 19 schools with a comprehensive school-based preventive oral health program, seven schools with a basic preventive program and six control schools. All sixth graders of these schools (n=1748) aged 11.7+/-1.1 years were clinically examined for black stain. DMFT was assessed in 1121 children by seven calibrated dentists using WHO criteria. DMFS was scored in 627 children by two calibrated dentists. Black stain was found in 16% of this population. The prevalence of black stain did not differ significantly between children attending schools with different oral health intervention programs. Thus, hypothesis 1 was accepted. The prevalence of black stain was significantly higher (Pcaries prevalence and caries experience than children without black stain. Thus, hypothesis 3 was rejected. No difference was found in the DMFS pattern of occlusal, smooth and proximal surfaces between children with and without black stain. Thus hypothesis 4 was accepted. The presence of black stain is

  5. Immunofluorescence Staining — EDRN Public Portal

    Science.gov (United States)

    Direct immunofluorescence method is used to detect the deposit of immunoglobulins, complement components, fibrinogen, etc. in tissues. This technique is usually performed on frozen sections. The primary antibody is conjugated to fluorescein binds directly with the antigen and can be detected by the fluorescent tag using a fluorescent microscope.

  6. 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.

  7. Wheat germ agglutinin staining as a suitable method for detection and quantification of fibrosis in cardiac tissue after myocardial infarction

    Directory of Open Access Journals (Sweden)

    B. Emde

    2014-10-01

    Full Text Available The quantification of fibrotic tissue is an important task in the analysis of cardiac remodeling. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. We therefore used FITC-labeled wheat germ agglutinin (WGA, which marks fibrotic tissue in comparable quality as the established picrosirius red (SR staining, for the staining of post myocardial infarction scar tissue. The fibrosis amount was quantified in a histogram-based approach using the non-commercial image processing program ImageJ. Our results clearly demonstrate that WGA-FITC is a suitable marker for cardiac fibrosis in frozen tissue sections. In combination with the histogram-based analysis, this new quantification approach is i easy and fast to perform; ii suitable for raw frozen tissue sections; and iii allows the use of additional antibodies in co-immunostaining. 

  8. 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.

  9. 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

  10. Hepatic involvement in paediatric patients with paracoccidioidomycosis: a histological study.

    Science.gov (United States)

    de Melo Braga, Giselle; Hessel, Gabriel; Pereira, Ricardo Mendes; Escanhoela, Cecília Amélia Fazzio

    2014-01-01

    Paracoccidioidomycosis is a systemic mycosis that is endemic to certain countries in Latin America. This study aimed to describe the histological features of liver involvement in patients with paracoccidioidomycosis aged <16 years of age who were treated between 1980 and 2010, with a diagnosis that was confirmed by detection of the fungus by pathological examination. Liver tissue was obtained from one necropsy and 12 biopsies. Throughout 2007, biopsies were taken from patients with persistent jaundice or portal hypertension, after which biopsies became indicated due to elevated aminotransferase and low albumin levels. Using haematoxylin and eosin (H&E), Masson's trichrome and immunohistochemical (CK7 and CK19) staining, we noted degenerative alterations in bile duct cells and inflammatory injury to the bile ducts in 10 biopsies. Using immunohistochemistry for CK7 and CK19, we observed ductal proliferation in all 12 samples. Bile duct injuries by inflammatory cells might explain the predominant increase in canalicular enzymes; immunohistochemistry is more sensitive in demonstrating ductular reactions and might show changes that are not apparent on H&E staining. © 2013 John Wiley & Sons Ltd.

  11. 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%.

  12. Histological characterization of lip and tentacle nerves in Lymnaea stagnalis.

    Science.gov (United States)

    Nakamura, H; Ito, I; Kojima, S; Fujito, Y; Suzuki, H; Ito, E

    1999-02-01

    The lip and tentacle nerves of the pond snail, Lymnaea stagnalis, were characterized using histological techniques. Anatomical drawings showed the detailed distributions of the superior lip, median lip, and tentacle nerves in the lip and tentacle; in particular it was found that the mouth is mainly innervated by the superior lip nerve. The tentacle nerve was clarified to form a zigzag structure along the extension direction in a shrinking tentacle. By backfilling of the superior lip nerve and/or the median lip nerve with fluorescent dyes, the neurons in the CNS made some clusters, whereas those stained from the tentacle nerve made other clusters. These stained neurons were not part of the central pattern generator or its regulatory neurons for feeding. The present results, therefore, suggest that the superior lip nerve may be employed as a principal factor in the chemosensory transduction from the mouth, and that no direct inputs occur through the lip and tentacle nerves to the central pattern generator or its regulatory neurons for feeding.

  13. Classification of breast cancer histology images using Convolutional Neural Networks.

    Science.gov (United States)

    Araújo, Teresa; Aresta, Guilherme; Castro, Eduardo; Rouco, José; Aguiar, Paulo; Eloy, Catarina; Polónia, António; Campilho, Aurélio

    2017-01-01

    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%.

  14. 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

  15. Keratocyte loss in corneal infection through apoptosis: a histologic study of 59 cases

    Directory of Open Access Journals (Sweden)

    Garg Prashant

    2004-12-01

    Full Text Available Abstract Background Keratocyte loss by apoptosis following epithelial debridement is a well-recognized entity. In a study of corneal buttons obtained from patients of corneal ulcer undergoing therapeutic keratoplasty, we observed loss of keratocytes in the normal appearing corneal stroma, surrounding the zone of inflammation. Based on these observations, we hypothesized that the cell loss in the inflammatory free zone of corneal stroma is by apoptosis that could possibly be a non-specific host response, independent of the nature of infectious agent. Methods To test our hypothesis, in this study, we performed Terminal deoxyribonucleotidyl transferase-mediated d-Uridine 5" triphosphate Nick End Labelling (TUNEL staining on 59 corneal buttons from patients diagnosed as bacterial, fungal, viral and Acanthamoeba keratitis. The corneal sections were reviewed for morphologic changes in the epithelium, stroma, type, degree and depth of inflammation, loss of keratocytes in the surrounding stroma (posterior or peripheral. TUNEL positivity was evaluated in the corneal sections, both in the zone of inflammation as well as the surrounding stroma. A correlation was attempted between the keratocyte loss, histologic, microbiologic and clinical features. Results The corneal tissues were from 59 patients aged between 16 years and 85 years (mean 46 years and included fungal (22, viral (15, bacterial (14 and Acanthamoeba (8 keratitis. The morphological changes in corneal tissues noted were: epithelial ulceration (52, 88.1%, destruction of Bowman's layer (58, 99%, mild to moderate (28; 47.5% to severe inflammation (31; 52.5%. Morphologic evidence of disappearance or reduced number of keratocytic nuclei in the corneal stroma was noted in 49 (83% cases; while the TUNEL positive brown cells were identified in all cases 53/54 (98%, including cases of fungal (19, bacterial (14, viral (13, and Acanthamoeba keratitis. TUNEL staining was located mostly in the deeper stroma

  16. 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.

  17. 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.

  18. Nucleus segmentation in histology images with hierarchical multilevel thresholding

    Science.gov (United States)

    Ahmady Phoulady, Hady; Goldgof, Dmitry B.; Hall, Lawrence O.; Mouton, Peter R.

    2016-03-01

    Automatic segmentation of histological images is an important step for increasing throughput while maintaining high accuracy, avoiding variation from subjective bias, and reducing the costs for diagnosing human illnesses such as cancer and Alzheimer's disease. In this paper, we present a novel method for unsupervised segmentation of cell nuclei in stained histology tissue. Following an initial preprocessing step involving color deconvolution and image reconstruction, the segmentation step consists of multilevel thresholding and a series of morphological operations. The only parameter required for the method is the minimum region size, which is set according to the resolution of the image. Hence, the proposed method requires no training sets or parameter learning. Because the algorithm requires no assumptions or a priori information with regard to cell morphology, the automatic approach is generalizable across a wide range of tissues. Evaluation across a dataset consisting of diverse tissues, including breast, liver, gastric mucosa and bone marrow, shows superior performance over four other recent methods on the same dataset in terms of F-measure with precision and recall of 0.929 and 0.886, respectively.

  19. Anolyte as an alternative bleach for stained cotton fabrics ...

    African Journals Online (AJOL)

    ... as the two- and three-factor interactions. The results from the study indicated that Anolyte was less effective than sodium hypochlorite as a stain remover for blood, tea, soot/mineral oil and blackcurrant juice. It was noted that the temperature of bleach liquids had an influence on the removal of stains by both bleach liquids.

  20. News from the Biological Stain Commission No. 10

    DEFF Research Database (Denmark)

    Lyon, H O

    2011-01-01

    In the 10th 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 meeting of ISO/TC 212/WG 1 held in London, UK, on 16-17 November 2009. Furthermore...

  1. 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...

  2. News from the Biological Stain Commission no. 13

    DEFF Research Database (Denmark)

    Lyon, H O

    2013-01-01

    In the 13(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 first plenary meeting of the International Standards Organization ISO/TC 212 Clinical lab...... laboratory testing and in vitro diagnostic test systems held on 17-19 October 2011 in Las Vegas, Nevada....

  3. 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...

  4. anolyte as an alternative bleach for stained cotton fabrics

    African Journals Online (AJOL)

    user

    Serowe College of Education. Private Bag 009. Serowe. Botswana ... and cons that may affect the environment as well as the quality of the final ..... improved as the pH became higher. Effects of anolyte, sodium hypochlorite and distilled water on the removal of tea stain on cotton. Leverette (2013:1) describes a tea stain as a.

  5. 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...

  6. 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.

  7. The second annular pulley: a histologic examination.

    Science.gov (United States)

    Ellis, F D; Seiler, J G; Sewell, C W

    1995-07-01

    The second annular pulley and underlying flexor tendons from 15 fresh frozen fingers were prepared, sectioned transversely and longitudinally and then stained with hematoxylin-eosin, Verhoeff-Van Gieson (Elastin), Masson's trichrome and for the presence of hyaluronic acid. Three discrete layers of the second annular pulley were identified (1) an outer layer, continuous with the membranous sheath, that was richly vascularized; (2) a middle layer, characterized by radially oriented collagen fibrils and significant elastin fibrils; (3) an inner gliding surface, characterized by longitudinally oriented collagen fibrils and modified fibroblasts secreting hyaluronic acid. The three layers of the pulley seem specialized and have developed based on the functional requirements of the pulley apparatus. The outer layer serves as a source of vascular supply and nutrition, the middle layer maintains structural integrity and is organized to efficiently resist flexor tendon bowstringing, and the innermost layer facilitates efficient excursion of the underlying flexor tendons both by the orientation of the collagen fibers and by the secretion of substances that are thought to minimize friction.

  8. [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.

  9. Diagnostic value of immunoperoxidase staining and immunofluorescence in the study of kidney biopsy specimens.

    Science.gov (United States)

    Jafari, Mohammad; Monsef-Esfahani, Alireza; Solimani, Bahram

    2015-07-01

    This study aimed to determine diagnostic value of immunoperoxidase in comparison with immunofluorescence in the diagnostic assessment of kidney biopsy specimens. Forty-eight kidney biopsy specimens were used to compare a direct immunofluorescence technique with immunoperoxidase techniques on paraffin sections. The sensitivity and specificity were calculated. The kappa statistic for agreement between the two tests was categorized as poor (zero to 0.2), moderate (0.21 to 0.45), good (0.46 to 0.75), and almost perfect concordance (0.76 to 1.0). Compared with immunofluorescence, the immunoperoxidase technique presented a sensitivity of 88.55% and a specificity of 69.22%. Its sensitivity in the staining for IgG, IgM, and IgA was 93.75%, 95.45%, and 76.47%, respectively. The specificity of this test in the staining for IgG, IgM, and IgA was 54.54%, 57.14%, and 96.00%, respectively. The overall kappa value was 0.60 and it was 0.60 for assessing staining intensity. There was a moderate agreement between immunoperoxidase and immunofluorescence in the positive or negative staining for IgG and IgM, as well as a good agreement in the positive or negative staining for IgA. For the staining intensity, the two tests had a good concordance for IgG and IgA and a moderate concordance for IgM. Although immunoperoxidase method has a lower overall diagnostic performance as compared to immunofluorescence, given the good concordance between the two techniques, it can be an alternative method for immunofluorescence study of kidney biopsy specimens, particularly where immunofluorescence fails or is not available.

  10. 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

  11. Remote histology learning from static versus dynamic microscopic images.

    Science.gov (United States)

    Mione, Sylvia; Valcke, Martin; Cornelissen, Maria

    2016-05-06

    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. Digitized microscopy images can be presented in either a static or dynamic format. This study of remote histology education identifies whether dynamic pictures are superior to static images for the acquisition of histological knowledge. Test results of two cohorts of second-year Bachelor in Medicine students at Ghent University were analyzed in two consecutive academic years: Cohort 1 (n = 190) and Cohort 2 (n = 174). Students in Cohort 1 worked with static images whereas students in Cohort 2 were presented with dynamic images. ANCOVA was applied to study differences in microscopy performance scores between the two cohorts, taking into account any possible initial differences in prior knowledge. The results show that practical histology scores are significantly higher with dynamic images as compared to static images (F (1,361) = 15.14, P Association of Anatomists. © 2015 American Association of Anatomists.

  12. Histological evaluation of the root apices of failed endodontic cases

    Directory of Open Access Journals (Sweden)

    Camilla Nicole Pecora

    2015-01-01

    Full Text Available Aim: The aim of this histological study was to evaluate the different causes of the failures of the endodontic treatment. Materials and Methods: Adult patients who were referred for endodontic surgery and for whom root-end resection was considered suitable were invited to participate in this study. The inclusion criteria were: (1 Patients aged between 18 and 65 years; (2 teeth with apical periodontitis that was diagnosed radiographically; (3 the tooth could not be adequately and better managed by root-canal retreatment; and (4 the crown of the tooth was adequately restored. One hundred root apices were surgically removed together with the periapical pathological tissue from 92 patients (56 males and 36 women.Histological sectioning was performed on calcified specimens that were embedded in a suitable medium. Results: The causes of endodontic failure identified through histological evaluation were as follows: Presence of bacterial and debris into canals (51%, apex transposition and overfilling (3%, presence of the isthmus (21%, bacterial colonization of root surface (2%, untreated canals (11%, and filling material outside root canal (4%. There was no significant difference between the distribution of teeth and the causes of endodontic failures (P = 0.32. Conclusions: The present study demonstrated that the most common cause of endodontic failure is the insufficient cleaning of the root canal system. It also explores the effectiveness of histological evaluation of the root apex following root-end resection in exploring the causes of endodontic failures.

  13. 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.

  14. A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. I. Optimalization of the staining procedure

    NARCIS (Netherlands)

    van Noorden, C. J.; Vogels, I. M.; James, J.; Tas, J.

    1982-01-01

    A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual human erythrocytes is described. This staining method can be used for the rapid routine discrimination of patients with a deficiency of the enzyme in its homozygote or heterozygote form, but also

  15. Hypoxia-inducible factor-2a is associated with ABCG2 expression, histology-grade and Ki67 expression in breast invasive ductal carcinoma

    Directory of Open Access Journals (Sweden)

    Xiang Lei

    2012-03-01

    Full Text Available Abstract Background Breast cancer is the most common cancer and the leading cause of cancer mortality in women worldwide. Hypoxia is an important factor involved in the progression of solid tumors and has been associated with various indicators of tumor metabolism, angiogenesis and metastasis. But little is known about the contribution of Hypoxia-Inducible Factor-2a (HIF-2a to the drug resistance and the clinicopathological characteristics in breast cancer. Methods Immunohistochemistry was employed on the tissue microarray paraffin sections of surgically removed samples from 196 invasive breast cancer patients with clinicopathological data. The correlations between the expression of HIF-2a and ABCG2 as well as other patients' clinicopathological data were investigated. Results The results showed that HIF-2a was expressed in different intensities and distributions in the tumor cells of the breast invasive ductal carcinoma. A positive staining for HIF-2a was defined as a brown staining observed mainly in the nucleus. A statistically significant correlation was demonstrated between HIF-2a expression and ABCG2 expression (p = 0.001, histology-grade (p = 0.029, and Ki67 (p = 0. 043 respectively. Conclusion HIF-2a was correlated with ABCG2 expression, histology-grade and Ki67 expression in breast invasive ductal carcinoma. HIF-2a could regulate ABCG2 in breast cancer cells, and could be a novel potential bio-marker to predict chemotherapy effectiveness. The hypoxia/HIF-2a/ABCG2 pathway could be a new mechanism of breast cancer multidrug-resistance. Virtual slides http://www.diagnosticpathology.diagnomx.eu/vs/2965948166714795

  16. 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.

  17. Novel Process for Laser Stain Removal from Archaeological Oil Paintings

    Science.gov (United States)

    El-Nadi, Lotfia; El-Feky, Osama; Abdellatif, Galila; Darwish, Sawsan

    2013-03-01

    Some samples of oil paintings (5 × 5 cm) were prepared on wooden panel with four types of fungi commonly encountered on oil paintings were selected for this study. Each of the fungi is associated with different colored stains. Fungus Alternaria tenuis is associated by a dense black stain, Chetomium globosum by a brownish gray stain, Aspergillus flavus by a yellowish stain, and Fusaruim oxysporum by a pinkish stain. Fungi growing on oil paintings affect the surface characteristics by forming a variety of colored patches typically composed of many complex chemical substances that are produced during metabolic processes. These colored stains may be encrusted in spores, present in mycelium or secreted to a substance such as oil paintings surfaces. While the fungal stains can sometimes be extracted with appropriate solvents, there are some stains that resist solvent extraction entirely. Developing new solvent system that might attack the paint structure, and is time consuming and requires a great deal of trial and error. Mechanical stain removal is also problematic in that it often produces abrasion of the surface, markedly deteriorating the artwork, and is extra ordinarily fine and tedious. For these reasons, we decided to examine an alternative physical technique as a new approach to deal with stain removal. Since the stains are due to the existence of fungi, we thought it a good idea to remove them by singlet oxygen. We applied the photo dynamic process through which the fungi stains were covered with organic dye derivatives in solution under controlled illumination in the lab. The samples were then irradiated by low power Laser light from a He-Ne laser, the dye will be photodecomposed and produce singlet oxygen. We report in this work the results obtained as a function of: - The concentration and types of the organic dye in solution, - The presence of certain amounts of liquids added to the solution, - The scanning speed of the laser beam on the sample surface

  18. 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

  19. vaginal histological changes of the baboon

    African Journals Online (AJOL)

    2009-04-04

    Apr 4, 2009 ... and endogenous (normal circulating hormones) on vaginal histology. In this study, we evaluated the vaginal histological changes in the baboon during the menstrual cycle. MATERIALS AND METHODS. Animal selection: Nine healthy adult female baboons. (Papio anubis) (mean±SD [15.2±2.03 Kg], range ...

  20. Rapid staining techniques in cytopathology: a review and comparison of modified protocols for hematoxylin and eosin, Papanicolaou and Romanowsky stains.

    Science.gov (United States)

    Jörundsson, Einar; Lumsden, John H.; Jacobs, Robert M.

    1999-01-01

    The objective of this study was to review and compare rapid protocols for fixation and staining of cytologic smears. We used fresh surgical specimens from dogs and horses to evaluate and modify, if necessary, previously described rapid staining protocols. Slides were wet-fixed, rehydrated or air-dried. Rapid Papanicolaou, hematoxylin and eosin (H&E), and Romanowsky stains were applied, including modification of Diff-Quick stain. The modified rapid staining protocols were simple to use and gave results within 5 minutes that were comparable to those obtained with traditional methods. Advantages of rehydrated vs wet-fixed smears included consistent preparations, a clean background, and equally good or superior nuclear detail.

  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. 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.

  3. 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.

  4. Microscopic analysis of MTT stained boar sperm cells

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2015-06-01

    Full Text Available The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI Stations is limited.

  5. Are there metallic traces in black extrinsic dental stain?

    Science.gov (United States)

    Parnas, Limor; Chevion, Mordechai; Berenshtein, Eduard; Faibis, Sarit; Moskovitz, Moti

    2013-05-01

    The detection of ferric ions in samples of black extrinsic dental stain led to the idea that it is comprised of insoluble ferric compounds. The present study examined the chemical composition of black extrinsic dental stain. Plaque was collected from 17 children with black extrinsic dental stain (study group A) and from 15 children without black extrinsic stain (control group), using sterile graphite curettes; and from 4 children with black extrinsic stain (study group B), using a standard sterile metal curette. Samples were analyzed with a scanning electron microscope (SEM) and subjected to quantitative chemical analysis (energy dispersive spectrometry). Except for calcium and phosphorus levels, no significant differences were found between the chemical composition of black extrinsic dental stain and dental plaque. Metallic ions were not detected in samples collected with a graphite curette (study group A), but were detected in samples collected with a metal curette (study group B). Metallic ions do not seem to be the origin of black extrinsic dental stain. Previous reports of the presence of metallic ions are probably due to contamination of the samples by the collection method.

  6. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

    Directory of Open Access Journals (Sweden)

    Andreyan Osipov

    2014-04-01

    Full Text Available This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977. The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine.

  7. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

    Science.gov (United States)

    Osipov, Andreyan; Arkhangelskaya, Ekaterina; Vinokurov, Alexei; Smetanina, Nadezhda; Zhavoronkov, Alex; Klokov, Dmitry

    2014-01-01

    This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. PMID:24727376

  8. 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.

  9. 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.

  10. 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.

  11. Diagnostic accuracy of cerebrospinal fluid gram stain in children with suspected bacterial meningitis.

    Science.gov (United States)

    Brizzi, Kate; Hines, Elizabeth M; McGowan, Karin L; Shah, Samir S

    2012-02-01

    This cross-sectional study included 1938 children undergoing lumbar puncture; 21 (1.1%) cases were classified as definite (n = 17) or probable (n = 4) bacterial meningitis. Gram stain sensitivity was 94.1% (95% confidence interval, 71.3%-99.9%) for those with definite meningitis; the positive predictive value was 47.1% (95% confidence interval, 29.8%-64.9%). The sensitivity was 95.2% for those with definite or probable meningitis. Antibiotic pretreatment did not affect results.

  12. 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...... of Regulatory Affairs, the Biological Stain Commission's International Affairs Committee presents information from the opening session of the meeting of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 2-4 June 2010 in Seoul, Republic...

  13. 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.

  14. Histological evaluation in ulcerative colitis

    Science.gov (United States)

    DeRoche, Tom C.; Xiao, Shu-Yuan; Liu, Xiuli

    2014-01-01

    This review summarizes diagnostic problems, challenges and advances in ulcerative colitis (UC). It emphasizes that, although histopathological examination plays a major role in the diagnosis and management of UC, it should always be interpreted in the context of clinical, endoscopic, and radiological findings. Accurate diagnosis requires knowledge of the classic morphological features of UC, as well as a number of atypical pathological presentations that may cause mis-classification of the disease process, either in resection or biopsy specimens. These atypical pathological presentations include rectal sparing and patchiness of disease at initial presentation of UC in pediatric patients or in the setting of medically treated UC, cecal or ascending colon inflammation in left-sided UC, and backwash ileitis in patients with severe ulcerative pancolitis. Loosely formed microgranulomas, with pale foamy histiocytes adjacent to a damaged crypt or eroded surface, should not be interpreted as evidence of Crohn’s disease. Indeterminate colitis should only be used in colectomy specimens as a provisional pathological diagnosis. Patients with UC are at risk for the development of dysplasia and carcinoma; optimal outcomes in UC surveillance programs require familiarity with the diagnostic criteria and challenges relating to UC-associated dysplasia and malignancy. Colon biopsy from UC patients should always be evaluated for dysplasia based on cytological and architectural abnormalities. Accurate interpretation and classification of dysplasia in colon biopsy from UC patients as sporadic adenoma or UC-related dysplasia [flat, adenoma-like, or dysplasia-associated lesion or mass (DALM)] requires clinical and endoscopic correlation. Isolated polypoid dysplastic lesions are considered to be sporadic adenoma if occurring outside areas of histologically proven colitis, or adenoma-like dysplasia if occurring in the diseased segment. Recent data suggest that such lesions may be treated

  15. Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining

    Directory of Open Access Journals (Sweden)

    Haixia Liu

    2017-01-01

    Full Text Available Aim. In routine histopathology, decalcification is an essential step for mineralized tissues. The purpose of this study is to evaluate the effects of different decalcification solutions on the morphological and antigenicity preservation in Sprague Dawley (SD rat femurs. Materials and Methods. Four different decalcification solutions were employed to remove the mineral substances from rat femurs, including 10% neutral buffered EDTA, 3% nitric acid, 5% nitric acid, and 8% hydrochloric acid/formic acid. Shaking and low temperature were used to process the samples. The stainings of hematoxylin-eosin (HE and immunohistochemical (IHC were employed to evaluate the bone morphology and antigenicity. Key Findings. Different decalcification solutions may affect the quality of morphology and the staining of paraffin-embedded sections in pathological examinations. Among four decalcifying solutions, 3% nitric acid is the best decalcifying agent for HE staining. 10% neutral buffered EDTA and 5% nitric acid are the preferred decalcifying agents for IHC staining. Significance. The current study investigated the effects of different decalcifying agents on the preservation of the bone structure and antigenicity, which will help to develop suitable protocols for the analyses of the bony tissue.

  16. Diffusion-weighted MR imaging in laryngeal and hypopharyngeal carcinoma: association between apparent diffusion coefficient and histologic findings.

    Science.gov (United States)

    Driessen, Juliette P; Caldas-Magalhaes, Joana; Janssen, Luuk M; Pameijer, Frank A; Kooij, Nina; Terhaard, Chris H J; Grolman, Wilko; Philippens, Marielle E P

    2014-08-01

    To investigate the relationship between the histologic characteristics of head and neck squamous cell carcinoma and apparent diffusion coefficient (ADC) at diffusion-weighted magnetic resonance (MR) imaging. The institutional ethics committee approved this study and waived informed consent. In head and neck squamous cell carcinoma, local failure after chemotherapy and/or radiation therapy correlates with pretreatment ADC. However, the histopathologic basis of this correlation remains unclear. In this study, 16 patients with head and neck squamous cell carcinoma were enrolled (median age, 60 years; range, 49-78 years). Before undergoing total laryngectomy, patients underwent 1.5-T diffusion-weighted MR imaging. After resection, whole-mount hematoxylin-eosin-stained sections were registered to the MR images. Cellular density; nuclear, cytoplasmic, and stromal area; and nuclear-cytoplasmic ratio within the tumor were calculated by using image-based segmentation on four consecutive slices. Mean ADC of the corresponding tumor region was calculated. Spearman correlations between ADC and histologic characteristics were calculated. ADC was significantly and inversely correlated with cell density (n = 16, r = -0.57, P = .02), nuclear area (n = 12, r = -0.64, P = .03), and nuclear-cytoplasmic ratio (n = 12, r = -0.77, P ≤ .01). ADC was significantly and positively correlated with percentage area of stroma (n = 12, r = 0.69, P = .01). Additionally, the percentage area of stroma was strongly interdependent with the percentage area of nuclei (n = 12, r = -0.97, P ≤ .01). ADC was significantly correlated with cellularity, stromal component, and nuclear-cytoplasmic ratio. The positive correlation of ADC and stromal component suggests that the poor prognostic value of high pretreatment ADC might partly be attributed to the tumor-stroma component, a known predictor of local failure.

  17. Protective effect of Concurrent Administration of Mesna and Galbanum Oil on Histological Structure of Rat Placenta against Cyclophosphamide Toxicity

    Directory of Open Access Journals (Sweden)

    Tayebeh Mohammadi

    2016-07-01

    Full Text Available Background & objectives: Cyclophosphamide is one of the common drugs used for cancer treatment. Cyclophosphamide affects the placenta and it is teratogenic in pregnant women. The purpose of this study was to evaluate the protective effects of Mesna and Galbanum essential oil concurrent administration against cyclophosphamide toxicity in rat placenta tissue. Methods: 22 pregnant female rats were randomly divided into 3 groups: (1 control (normal saline, n=7 rats, (2 cyclophosphamide (15mg/kg, n=7 rats, (3 cyclophosphamide (15mg/kg, Mesna (100mg/kg and Galbanum (200mg/kg, n=8 rats. Medications were administered intraperitoneally on the 13th day of pregnancy. Then, rats were sacrificed on the 20th day of pregnancy. The placenta was separated and fixed after macroscopic evaluation. 5 µm tissue sections were prepared, stained and studied by light microscopy. Results: Cyclophosphamide reduced the placental weight, thickness and the length of the small and large diameters. Histologically, it reduced the thickness of the labyrinth and basal layers, caused necrosis and congestion in these layers, reduced giant cells numbers and caused their nuclear shrinkage (p0.05 but histologically it could increase the labyrinth and basal layers thickness and also the giant cells numbers (p<0.05. It could also prevent nuclear shrinkage, giant cells and spongiotrophoblasts necrosis in the basal layer and reduce congestion in the labyrinth layer (p<0.05. Conclusion: Mesna and Galbanum oil preserved the placenta of pregnant rat against toxic effects of cyclophosphamide probably by its antioxidant features and free radicals deletion.

  18. 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.

  19. Histological evaluation of the biocompatibility of polyurea crosslinked silica aerogel implants in a rat model: a pilot study.

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    Firouzeh Sabri

    Full Text Available BACKGROUND: Aerogels are a versatile group of nanostructured/nanoporous materials with physical and chemical properties that can be adjusted to suit the application of interest. In terms of biomedical applications, aerogels are particularly suitable for implants such as membranes, tissue growth scaffolds, and nerve regeneration and guidance inserts. The mesoporous nature of aerogels can also be used for diffusion based release of drugs that are loaded during the drying stage of the material. From the variety of aerogels polyurea crosslinked silica aerogels have the most potential for future biomedical applications and are explored here. METHODOLOGY: This study assessed the short and long term biocompatibility of polyurea crosslinked silica aerogel implants in a Sprague-Dawley rat model. Implants were inserted at two different locations a subcutaneously (SC, at the dorsum and b intramuscularly (IM, between the gluteus maximus and biceps femoris of the left hind extremity. Nearby muscle and other internal organs were evaluated histologically for inflammation, tissue damage, fibrosis and movement (travel of implant. CONCLUSION/SIGNIFICANCE: In general polyurea crosslinked silica aerogel (PCSA was well tolerated as a subcutaneous and an intramuscular implant in the Sprague-Dawley rat with a maximum incubation time of twenty months. In some cases a thin fibrous capsule surrounded the aerogel implant and was interpreted as a normal response to foreign material. No noticeable toxicity was found in the tissues surrounding the implants nor in distant organs. Comparison was made with control rats without any implants inserted, and animals with suture material present. No obvious or noticeable changes were sustained by the implants at either location. Careful necropsy and tissue histology showed age-related changes only. An effective sterilization technique for PCSA implants as well as staining and sectioning protocol has been established. These studies

  20. 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.

  1. Methylene Blue: The Long and Winding Road from Stain to Brain: Part 1.

    Science.gov (United States)

    Howland, Robert H

    2016-09-01

    Methylene blue, first discovered and used as a dye in the textile industry, has long been used for biological staining in histology, bacteriology, and hematology. Because of its unique physiochemical properties, it was the first synthetic drug used in medicine, having been used to treat malaria more than one century ago. Methylene blue was also one of the first drugs used for the treatment of patients with psychosis at the end of the 19th century and was the lead drug in the serendipitous development of phenothiazine antipsychotic drugs in the mid-20th century. It was studied in bipolar disorder in the 1980s and has been investigated in neurodegenerative disorders in recent years. The history of methylene blue from its discovery as a dye to its use as a stain and then its therapeutic application in medicine is an example of how a drug's use can evolve over time through careful observation, clinical needs, serendipity, and the integration of concepts from different disciplines. [Journal of Psychosocial Nursing and Mental Health Services, 54(9), 21-24.]. Copyright 2016, SLACK Incorporated.

  2. The Gram stain after more than a century.

    Science.gov (United States)

    Popescu, A; Doyle, R J

    1996-05-01

    The Gram stain, the most important stain in microbiology, was described more than a century ago. Only within the past decade, however, has an understanding of its mechanism emerged. It now seems clear that the cell wall of Gram-positive microorganisms is responsible for retention of a crystal violet:iodine complex. In Gram-negative cells, the staining procedures damage the cell surface resulting in loss of dye complexes. Gram-positive microorganisms require a relatively thick cell wall, irrespective of composition, to retain the dye. Therefore, Gram-stainability is a function of the cell wall and is not related to chemistry of cell constituents. This review provides a chronology of the Gram stain and discusses its recently discovered mechanism.

  3. Standardization of the Romanowsky staining procedure: an overview.

    Science.gov (United States)

    Bentley, S A; Marshall, P N; Trobaugh, F E

    1980-01-01

    Commerically available Romanowsky blood stains are variable mixtures of thiazein dyes and brominated fluorescein derivatives with varying degrees of metallic salt contamination in a number of different solvent systems. There is a need for standardized Romanowsky stains of constant composition, which, when used in conjunction with a carefully controlled specimen preparation technique, should give consistent performance. Such a preparation system would be of great value to hematologists in general and would be essential to the validity of data obtained by the digital processing of blood cell images. It is possible to prepare standardized Romanowsky stains as mixtures of two or three dye components, namely, eosin Y, azure B and methylene blue, although azure B has only recently become commercially available at an acceptable degree of purity. The logistic problems of stain standardization are discussed.

  4. The value of intraoperative Gram stain in revision spine surgery.

    Science.gov (United States)

    Shifflett, Grant D; Nwachukwu, Benedict U; Bjerke-Kroll, Benjamin T; Kueper, Janina; Koltsov, Jayme B; Sama, Andrew A; Girardi, Federico P; Cammisa, Frank P; Hughes, Alexander P

    2015-10-01

    Intraoperative cultures and Gram stains are often obtained in cases of revision spine surgery even when clinical signs of infection are not present. The clinical utility and cost-effectiveness of this behavior remain unproven. The aim was to evaluate the clinical utility and cost-effectiveness of routine intraoperative Gram stains in revision spine surgery. This was a retrospective clinical review performed at an academic center in an urban setting. One hundred twenty-nine consecutive adult revision spine surgeries were performed. The outcome measures included intraoperative Gram stains. We retrospectively reviewed the records of 594 consecutive revision spine surgeries performed by four senior surgeons between 2008 and 2013 to identify patients who had operative cultures and Gram stains performed. All revision cases including cervical, thoracic, and lumbar fusion and non-fusion, with and without instrumentation were reviewed. One hundred twenty-nine (21.7%) patients had operative cultures obtained and were included in the study. The most common primary diagnosis code at the time of revision surgery was pseudarthrosis, which was present in 41.9% of cases (54 of 129). Infection was the primary diagnosis in 10.1% (13 of 129) of cases. Operative cultures were obtained in 129 of 595 (21.7%) cases, and 47.3% (61 of 129) were positive. Gram stains were performed in 98 of 129 (76.0%) cases and were positive in 5 of 98 (5.1%) cases. Overall, there was no correlation between revision diagnosis and whether or not a Gram stain was obtained (p=.697). Patients with a history of prior instrumentation were more likely to have a positive Gram stain (pGram staining was found to have a sensitivity of 10.9% (confidence interval [CI] 3.9%-23.6%) and specificity of 100% (CI 93.1%-100%). The positive and negative predictive values were 100% (CI 48.0%-100%) and 57.3% (CI 45.2%-66.2%), respectively. Kappa coefficient was calculated to be 0.1172 (CI 0.0194-0.2151). The cost per discrepant

  5. 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

  6. 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.

  7. NEONATAL OUTCOME IN MECONIUM STAINED DELIVERIES — A PROSPECTIVE STUDY

    OpenAIRE

    RAMAN, TS RAGHU; JAYAPRAKASH, DG

    1997-01-01

    This prospective study analyzes the neonatal outcome in deliveries complicated by meconium stained amniotic fluid. In a study of 1000 live born deliveries, meconium staining of amniotic fluid was seen in 50 (5%) deliveries. Out of these, 20 newborns (40%) developed classical signs of meconium aspiration syndrome and were managed according to a predetermined protocol. Multiparity, term deliveries, use of sedatives in mother, intrauterine growth retardation and prolonged labour were some of the...

  8. News from the Biological Stain Commission no. 15

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2014-01-01

    In the 15(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 plenary meetings of the International Standards Organization ISO/TC 212 Clinical laborat...... laboratory testing and in vitro diagnostic test systems held on August 22-24, 2012 in Berlin, Germany. An additional discussion of the use of food dyes in India also is included....

  9. 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.

  10. Effects of body weight and nutrition on histological mammary development in Holstein heifers.

    Science.gov (United States)

    Daniels, K M; McGilliard, M L; Meyer, M J; Van Amburgh, M E; Capuco, A V; Akers, R M

    2009-02-01

    Our objective was to determine the effects of rate of gain and body weight (BW) on development of the mammary parenchyma. Mammary tissue samples were collected from heifers (n = 72) reared on 1 of 2 dietary treatments (restricted, 650 g/d of daily gain; or elevated, 950 g/d of daily gain) and slaughtered at 100, 150, 200, 250, 300, or 350 kg of BW. Mammary samples were excised, preserved, prepared for histology, and stained with hematoxylin and eosin. Digital images of tissue sections were captured for analysis. Tissue areas occupied by the interlobular and intralobular stroma, epithelium, and lumen were measured (mum(2)). The numbers of epithelial and luminal structures per image were tabulated to measure the complexity of ductal development. Mean percentages of mammary parenchyma occupied by the interlobular stroma, epithelium, lumen, and intralobular stroma were 29, 20, 7, and 43%, respectively. Percentage of area occupied by the intralobular stroma was affected by BW and was lower for 100-kg heifers compared with heifers 200 kg and heavier (33 +/- 4 vs. 46 +/- 4), but the percentage of area occupied by other tissue elements did not differ by BW or treatment, nor was there an interaction. However, the numbers of both epithelial (8.3 +/- 4 vs. 47 +/- 4) and luminal-containing (6 +/- 4 vs. 38 +/- 4) structures per image increased markedly between 100 and 350 kg of BW, irrespective of diet. For heifers slaughtered between 100 and 350 kg of BW, alterations in the rate of gain between 650 and 950 g/d, accomplished by feeding varying amounts of the same diet, had no significant effect on tissue characteristics or the pattern of mammary parenchymal development. These data emphasize the importance of BW and age in determining developmental characteristics of the heifer mammary parenchyma and suggest that the rate of gain per se has a minimal impact on histological development, and thus do not support the hypothesis that rate of gain has a direct negative impact on

  11. 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

  12. A histological procedure to study fungal infection in the wax moth Galleria mellonella

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    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.

  13. A histological procedure to study fungal infection in the wax moth Galleria mellonella.

    Science.gov (United States)

    Perdoni, F; Falleni, M; Tosi, D; Cirasola, D; Romagnoli, S; Braidotti, P; Clementi, E; Bulfamante, G; Borghi, E

    2014-09-23

    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.

  14. Histological validation of high-resolution DTI in human post mortem tissue.

    Science.gov (United States)

    Seehaus, Arne; Roebroeck, Alard; Bastiani, Matteo; Fonseca, Lúcia; Bratzke, Hansjürgen; Lori, Nicolás; Vilanova, Anna; Goebel, Rainer; Galuske, Ralf

    2015-01-01

    Diffusion tensor imaging (DTI) is amongst the simplest mathematical models available for diffusion magnetic resonance imaging, yet still by far the most used one. Despite the success of DTI as an imaging tool for white matter fibers, its anatomical underpinnings on a microstructural basis remain unclear. In this study, we used 65 myelin-stained sections of human premotor cortex to validate modeled fiber orientations and oft used microstructure-sensitive scalar measures of DTI on the level of individual voxels. We performed this validation on high spatial resolution diffusion MRI acquisitions investigating both white and gray matter. We found a very good agreement between DTI and myelin orientations with the majority of voxels showing angular differences less than 10°. The agreement was strongest in white matter, particularly in unidirectional fiber pathways. In gray matter, the agreement was good in the deeper layers highlighting radial fiber directions even at lower fractional anisotropy (FA) compared to white matter. This result has potentially important implications for tractography algorithms applied to high resolution diffusion MRI data if the aim is to move across the gray/white matter boundary. We found strong relationships between myelin microstructure and DTI-based microstructure-sensitive measures. High FA values were linked to high myelin density and a sharply tuned histological orientation profile. Conversely, high values of mean diffusivity (MD) were linked to bimodal or diffuse orientation distributions and low myelin density. At high spatial resolution, DTI-based measures can be highly sensitive to white and gray matter microstructure despite being relatively unspecific to concrete microarchitectural aspects.

  15. Diagnostic capabilities of high-definition white light endoscopy for the diagnosis of gastric intestinal metaplasia and correlation with histologic and clinical data.

    Science.gov (United States)

    Panteris, Vasileios; Nikolopoulou, Stamatina; Lountou, Anastasia; Triantafillidis, John K

    2014-06-01

    The aim of this study was the evaluation of the diagnostic accuracy of a specific high-definition white light endoscopy (HD-WLE) system for the optical recognition of intestinal metaplasia (IM) and the assessment of its correlation with histologic and clinical data. A total of 234 patients undergoing upper gastrointestinal endoscopy in an outpatient endoscopy suite for various indications were prospectively enrolled in this cross-sectional study. Gastric IM was diagnosed on the basis of three mucosal patterns identified using HD-WLE in a per-patient analysis. Histological evaluation was used as the gold standard, and special staining was conducted for subtyping of IM. Main outcome measurements were sensitivity, specificity, and likelihood ratio of HD-WLE and secondary associations with histologic and clinical data. IM was found in 63/234 (27%) patients and low-grade dysplasia in 6/63 patients (9.5%). Sensitivity, specificity, accuracy, and likelihood ratio of all mucosal patterns were 74.6, 94, 88% and 13, respectively. All clinically significant type III IM and dysplasia lesions were endoscopically detected. All nonvisible lesions were of types I and II with mild grade and no dysplasia. Ten patients were considered false positives and the lesions were associated with severe inflammation and antralization. The specific HD-WLE system showed satisfactory accuracy and high specificity during real-time, routine endoscopy practice. Specific mucosal patterns were correlated with level and grade of lesions. The sensitivity of the system is even higher when only clinically significant IM lesions are considered.

  16. 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.

  17. 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...

  18. 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 < .01). Post-hoc analysis reveals that polished specimens had significantly less color change than un-polished specimens for Paradigm C and LAVA Ultimate. E.max had significantly higher 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.

  19. Mixed adenosquamous histology is associated with poorer survival of cervical cancer stage 1b

    DEFF Research Database (Denmark)

    Lauszus, Finn; Tjessem, Ingvild; Al-Far, Hanine M

    2016-01-01

    , and post mortem sections provided follow-up on survival, relapse, and re-admittance. Histopathological evaluation was performed by the same pathologist. Results: Histological evaluation showed that adenosquamous cervical cancer in stage 1b was associated with poorer survival than the pure squamous......, while survival was found to be 89 % (83-95 %) and 86 % (79-93 %), respectively. The women’s age at diagnosis showed no association with histology type or survival....

  20. Histological Types of Polypoid Cutaneous Melanoma II

    OpenAIRE

    Knežević, Fabijan; Duančić, Vjekoslav; Šitić, Sanda; Horvat-Knežević, Anica; Benković, Vesna; Ramić, Snježana; Kostović, Krešimir; Ramljak, Vesna; Velemir Vrdoljak, Danko; Stanec, Mladen; Božović, Angelina

    2007-01-01

    The aim of this study was to ascertain which histological types of melanoma can clinically and morphologically appear as polypoid melanomas. In 645 cases of primary cutaneous melanoma we have analyzed criteria for diagnosis of polypoid cutaneous melanoma and afterwards we have analyzed growth phase in each polypoid melanoma, histological type of atypical melanocytes, the number of epidermal ridges which are occupied by atypical melanocytes, and distribution according to age, sex a...

  1. Histological observations on Montenegro's reaction in man

    OpenAIRE

    Mayrink, Wilson; Schettini, Antonio Pedro M.; Williams, Paul; Raso, Pedro; Magalhães, Paulo Araujo; Lima, Antonio De Oliveira; Melo, Maria Norma; Costa, Carlos Alberto da; Genaro, Odair; Dias, Magno; Michalick, Marilene Suzan Marques

    1989-01-01

    The Montenegro skin test is widely used as a diagnostic method for American cutaneous leishmaniasis (ACL) but little is known about the histological changes that occur in the skin after administration of the antigen. This report is based on histological studies of biopsied material obtained, from inoculation sites, 48 hours after individuals had been given intradermal injections with a standardized Montenegro antigen. The material examined was obtained from four distinctly different test grou...

  2. GRAPHIE: graph based histology image explorer.

    Science.gov (United States)

    Ding, Hao; Wang, Chao; Huang, Kun; Machiraju, Raghu

    2015-01-01

    Histology images comprise one of the important sources of knowledge for phenotyping studies in systems biology. However, the annotation and analyses of histological data have remained a manual, subjective and relatively low-throughput process. We introduce Graph based Histology Image Explorer (GRAPHIE)-a visual analytics tool to explore, annotate and discover potential relationships in histology image collections within a biologically relevant context. The design of GRAPHIE is guided by domain experts' requirements and well-known InfoVis mantras. By representing each image with informative features and then subsequently visualizing the image collection with a graph, GRAPHIE allows users to effectively explore the image collection. The features were designed to capture localized morphological properties in the given tissue specimen. More importantly, users can perform feature selection in an interactive way to improve the visualization of the image collection and the overall annotation process. Finally, the annotation allows for a better prospective examination of datasets as demonstrated in the users study. Thus, our design of GRAPHIE allows for the users to navigate and explore large collections of histology image datasets. We demonstrated the usefulness of our visual analytics approach through two case studies. Both of the cases showed efficient annotation and analysis of histology image collection.

  3. 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.

  4. 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

  5. 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.

  6. The influence of Romanowsky-Giemsa type stains on nuclear and cytoplasmic features of cytological specimens.

    Science.gov (United States)

    Schulte, E; Wittekind, D

    1989-04-01

    The aim of the present study was to compare the staining pattern of the standard azure B-eosin Y stain with commercial May-Grünwald-Giemsa (MGG) stains on cytological specimens by means of high resolution image analysis. Several cytological specimens (blood smears, abdominal serous effusions, bronchial scrape material) were air dried, methanol fixed and stained with the standard azure B-eosin Y stain and with commercial May-Grünwald-Giemsa stains. Integrated optical density (IOD) and colour intensities of cell nuclei and cytoplasm were measured with the IBAS 2000 image analyser. Commercial MGG stains gave much higher coefficients of variation for all parameters than the standard stain. Reproducibility of cell nuclei segmentation versus cytoplasm was significantly better for the standard stain. Contamination of the standard stain with methylene blue partly copied the staining pattern of commercial stains. The standard azure B-eosin Y stain is recommended for high resolution image analysis (HRIA) of cytological samples.

  7. 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.

  8. Leishman-Giemsa Cocktail - Is it an Effective Stain for Air Dried Cytology Smears.

    Science.gov (United States)

    Doddagowda, Shilpa Manigatta; Shashidhar, Hemalatha Anantharamaiah; Prasad, Chinaiah Subramanyam Babu Rajendra

    2017-03-01

    Air dried cytology smears are stained routinely with Romanowsky stains so that the relative cell size, nuclear size, cytoplasmic details, smear background elements and intercellular matrix components are better appreciated. A variety of modified Romanowsky stains are used in cytology. Leishman-Giemsa (LG) cocktail is one of the new staining techniques which can be used for staining the air dried cytology smears. To evaluate the quality of staining of LG cocktail on air dried smears and to compare the quality of staining of LG cocktail with May Grunwald Giemsa (MGG) which is the most commonly used stain in cytology. The present prospective comparative study was carried out with 100 cases and two extra smears were prepared for each case and stained with MGG and LG cocktail stains. The stained slides were blinded and were evaluated for the staining characteristics of the nucleus, cytoplasm and background staining. Based on this, scoring was done by two pathologists independently. Quality Index (QI) was calculated by dividing the scores obtained with the total score possible. LG cocktail stained slides were excellent in cytoplasmic staining, granularity, nuclear morphology, background material staining and overall staining characteristics. QI of LG cocktail was 0.8 while that of MGG was 0.59. Staining of air dried smears by LG cocktail has a good QI. It is also cheaper, requires short duration for staining as compared to MGG. Hence, LG cocktail can be an effective replacement for MGG for staining the air dried cytology smears.

  9. Rinsing paired-agent model (RPAM) to quantify cell-surface receptor concentrations in topical staining applications of thick tissues

    Science.gov (United States)

    Xu, Xiaochun; Wang, Yu; Xiang, Jialing; Liu, Jonathan T. C.; Tichauer, Kenneth M.

    2017-06-01

    Conventional molecular assessment of tissue through histology, if adapted to fresh thicker samples, has the potential to enhance cancer detection in surgical margins and monitoring of 3D cell culture molecular environments. However, in thicker samples, substantial background staining is common despite repeated rinsing, which can significantly reduce image contrast. Recently, ‘paired-agent’ methods—which employ co-administration of a control (untargeted) imaging agent—have been applied to thick-sample staining applications to account for background staining. To date, these methods have included (1) a simple ratiometric method that is relatively insensitive to noise in the data but has accuracy that is dependent on the staining protocol and the characteristics of the sample; and (2) a complex paired-agent kinetic modeling method that is more accurate but is more noise-sensitive and requires a precise serial rinsing protocol. Here, a new simplified mathematical model—the rinsing paired-agent model (RPAM)—is derived and tested that offers a good balance between the previous models, is adaptable to arbitrary rinsing-imaging protocols, and does not require calibration of the imaging system. RPAM is evaluated against previous models and is validated by comparison to estimated concentrations of targeted biomarkers on the surface of 3D cell culture and tumor xenograft models. This work supports the use of RPAM as a preferable model to quantitatively analyze targeted biomarker concentrations in topically stained thick tissues, as it was found to match the accuracy of the complex paired-agent kinetic model while retaining the low noise-sensitivity characteristics of the ratiometric method.

  10. Polymorphous low-grade adenocarcinoma of minor salivary gland. A comparative histologic and immunohistochemical study.

    Science.gov (United States)

    Regezi, J A; Zarbo, R J; Stewart, J C; Courtney, R M

    1991-04-01

    Sixteen polymorphous low-grade adenocarcinomas were reviewed and compared with 17 adenoid cystic carcinomas and with 21 other histologically similar minor salivary gland neoplasms. The polymorphous low-grade adenocarcinomas were for the most part distinctive in their microscopic appearance. Typically they exhibited infiltrative growth by small uniform cells in single-layered ducts. A syncytium of tumor cells was also characteristic, although solid and cribriform patterns were seen, making definitive diagnosis difficult with some tumors. Immunohistochemical staining for S-100 protein, glial fibrillary acidic protein, actin, vimentin, and keratins resulted in relatively distinctive antigenic profiles for the tumors studied. Of significance was strong S-100 protein and weak actin staining of polymorphous low-grade adenocarcinomas, moderate actin staining of adenoid cystic carcinomas, moderate glial fibrillary acidic protein staining of monomorphic adenomas and pleomorphic adenomas, and nonreactivity of monomorphic adenomas for vimentin. It is believed that the immunoprofiles could be useful in the microscopic diagnosis of salivary gland tumors. The identification of antigens found normally in myoepithelial and epithelial cells supports the concept that these tumors are derived from pluripotential reserve cells.

  11. Technique for histological control of surgical margins in lip cancer.

    Science.gov (United States)

    Aoyagi, Satoru; Hata, Hiroo; Homma, Erina; Shimizu, Hiroshi

    2014-04-01

    To preserve oral function and achieve acceptable cosmetic results, intraoperative control of surgical margins with frozen section evaluation may help to determine surgical technique in lip cancer. However, frozen section analysis is usually limited to suspicious areas and has not been systematically performed among surgeons. The accuracy of such analysis for detecting histological surgical margins is highly dependent on the methods used to obtain and analyze the margins. Improving the pathodiagnostic reliability of conventional intraoperative frozen section evaluation is the most important goal of surgical management in our method. We describe the successful use of the "double-blade method" in lip cancer treatment. The technique we describe has the advantage of histologically confirming clear margins in lip cancer. This method appears to be time-saving and easy to apply with existing surgical systems. In addition, this method may be used as an alternative to complete evaluation of lateral surgical margins that is important in planning a suitable surgical reconstruction procedure in lip cancer at many institutions where Mohs micrographic surgery is difficult to perform. © 2014 Japanese Dermatological Association.

  12. A staining protocol for identifying secondary compounds in Myrtaceae1

    Science.gov (United States)

    Retamales, Hernan A.; Scharaschkin, Tanya

    2014-01-01

    • Premise of the study: Here we propose a staining protocol using toluidine blue (TBO) and ruthenium red to reliably identify secondary compounds in the leaves of some species of Myrtaceae. • Methods and Results: Leaves of 10 species representing 10 different genera of Myrtaceae were processed and stained using five different combinations of ruthenium red and TBO. Optimal staining conditions were determined as 1 min of ruthenium red (0.05% aqueous) and 45 s of TBO (0.1% aqueous). Secondary compounds clearly identified under this treatment include mucilage in the mesophyll, polyphenols in the cuticle, lignin in fibers and xylem, tannins and carboxylated polysaccharides in the epidermis, and pectic substances in the primary cell walls. • Conclusions: Potential applications of this protocol include systematic, phytochemical, and ecological investigations in Myrtaceae. It might be applicable to other plant families rich in secondary compounds and could be used as a preliminary screening method for extraction of these elements. PMID:25309840

  13. A staining protocol for identifying secondary compounds in Myrtaceae.

    Science.gov (United States)

    Retamales, Hernan A; Scharaschkin, Tanya

    2014-10-01

    Here we propose a staining protocol using toluidine blue (TBO) and ruthenium red to reliably identify secondary compounds in the leaves of some species of Myrtaceae. • Leaves of 10 species representing 10 different genera of Myrtaceae were processed and stained using five different combinations of ruthenium red and TBO. Optimal staining conditions were determined as 1 min of ruthenium red (0.05% aqueous) and 45 s of TBO (0.1% aqueous). Secondary compounds clearly identified under this treatment include mucilage in the mesophyll, polyphenols in the cuticle, lignin in fibers and xylem, tannins and carboxylated polysaccharides in the epidermis, and pectic substances in the primary cell walls. • Potential applications of this protocol include systematic, phytochemical, and ecological investigations in Myrtaceae. It might be applicable to other plant families rich in secondary compounds and could be used as a preliminary screening method for extraction of these elements.

  14. 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 number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay...... may itself kill sperm, which is likely to confound many common experimental designs in addition to producing artificially low estimates of sperm viability. I further suggest that sperm number should be routinely measured in sperm viability studies, as it may be an important but overlooked source...

  15. Vital staining of coronal dentin in monkey teeth.

    Science.gov (United States)

    Tronstad, L

    1978-04-01

    Vital staining of monkey incisor teeth with the incisal dentin exposed to the oral environment by attrition was carried out, with the use of a number of dyes (pH and redox indicators). There was a distinct staining of the coronal dentin, regardless of which dye was introduced into the pulpal cavity. The exposed dentin was stained like the unaffected dentin, with the exception of a narrow centrally located zone that extended from the tip of the original pulp horn to the incisal edge of the tooth. The suggestion is that this zone is not unstained because of exposure of the dentin to the oral environment, but because it coincides with an area of the tissue where the pulpal ends of the dentinal tubules are blocked by atubular hard tissue normally laid down in the pulp horn of incisor teeth.

  16. 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.

  17. Quantitative histological validation of diffusion MRI fiber orientation distributions in the rat brain.

    Science.gov (United States)

    Leergaard, Trygve B; White, Nathan S; de Crespigny, Alex; Bolstad, Ingeborg; D'Arceuil, Helen; Bjaalie, Jan G; Dale, Anders M

    2010-01-07

    Diffusion MRI (dMRI) is widely used to measure microstructural features of brain white matter, but commonly used dMRI measures have limited capacity to resolve the orientation structure of complex fiber architectures. While several promising new approaches have been proposed, direct quantitative validation of these methods against relevant histological architectures remains missing. In this study, we quantitatively compare neuronal fiber orientation distributions (FODs) derived from ex vivo dMRI data against histological measurements of rat brain myeloarchitecture using manual recordings of individual myelin stained fiber orientations. We show that accurate FOD estimates can be obtained from dMRI data, even in regions with complex architectures of crossing fibers with an intrinsic orientation error of approximately 5-6 degrees in these regions. The reported findings have implications for both clinical and research studies based on dMRI FOD measures, and provide an important biological benchmark for improved FOD reconstruction and fiber tracking methods.

  18. Quantitative histological validation of diffusion MRI fiber orientation distributions in the rat brain.

    Directory of Open Access Journals (Sweden)

    Trygve B Leergaard

    2010-01-01

    Full Text Available Diffusion MRI (dMRI is widely used to measure microstructural features of brain white matter, but commonly used dMRI measures have limited capacity to resolve the orientation structure of complex fiber architectures. While several promising new approaches have been proposed, direct quantitative validation of these methods against relevant histological architectures remains missing. In this study, we quantitatively compare neuronal fiber orientation distributions (FODs derived from ex vivo dMRI data against histological measurements of rat brain myeloarchitecture using manual recordings of individual myelin stained fiber orientations. We show that accurate FOD estimates can be obtained from dMRI data, even in regions with complex architectures of crossing fibers with an intrinsic orientation error of approximately 5-6 degrees in these regions. The reported findings have implications for both clinical and research studies based on dMRI FOD measures, and provide an important biological benchmark for improved FOD reconstruction and fiber tracking methods.

  19. Choroid plexus papilloma of the third ventricle: angiography, preoperative embolization, and histology

    Energy Technology Data Exchange (ETDEWEB)

    Do, H.M. [Interventional Neuroradiology, S-047, Department of Radiology, Stanford University Medical Center (United States); Dept. of Radiology, University of Virginia Health Sciences Center, Charlottesville, VA (United States); Marx, W.F. [Dept. of Radiology, University of Virginia Health Sciences Center, Charlottesville, VA (United States); Khanam, H. [Dept. of Pathology, University of Virginia Health Sciences Center, Charlottesville, VA (United States); Jensen, M.E. [Dept. of Radiology, University of Virginia Health Sciences Center, Charlottesville, VA (United States); Dept. of Neurosurgery, University of Virginia Health Sciences Center, Charlottesville, VA (United States)

    2001-06-01

    We report a unique case of choroid plexus papilloma of the third ventricle in an 8-month-old girl in which preoperative embolization played a salient role in management. Initial surgery was aborted due to excessive bleeding. Cerebral angiography demonstrated enlarged posterior choroidal arteries feeding the tumor, and intense, persistent tumor staining. These vessels were effectively embolized to stasis with polyvinyl alcohol particles. The patient underwent a second craniotomy and complete resection of the tumor with minimal blood loss. Postsurgical histology showed postembolization iatrogenic intratumoral necrosis. (orig.)

  20. Intestinal Bacterial Infection Diagnosed by Histological Examination of Endoscopic Biopsy Specimens

    Directory of Open Access Journals (Sweden)

    Michiko Yuki

    2016-10-01

    Full Text Available Intestinal spirochetosis (IS in humans is characterized by spirochetal microorganisms attached to the luminal surface of the colonic epithelium. In the present case, attached organisms appeared as 3- to 4 μm-thick (average thickness, 3.4 μm basophilic fringes or haze in HE-stained endoscopic biopsy specimens. The basophilic fringes were clearly labeled by Treponema pallidum antiserum. Because IS is relatively rare in developed countries, thin basophilic fringes characteristic of IS are readily overlooked. Thus, the recognition of histological characteristics of this disease is important for its diagnosis.