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Sample records for enhance u-par staining

  1. Inhibition of uPAR-TGFβ crosstalk blocks MSC-dependent EMT in melanoma cells.

    Science.gov (United States)

    Laurenzana, Anna; Biagioni, Alessio; Bianchini, Francesca; Peppicelli, Silvia; Chillà, Anastasia; Margheri, Francesca; Luciani, Cristina; Pimpinelli, Nicola; Del Rosso, Mario; Calorini, Lido; Fibbi, Gabriella

    2015-07-01

    The capacity of cancer cells to undergo epithelial-to-mesenchymal transition (EMT) is now considered a hallmark of tumor progression, and it is known that interactions between cancer cells and mesenchymal stem cells (MSCs) of tumor microenvironment may promote this program. Herein, we demonstrate that MSC-conditioned medium (MSC-CM) is a potent inducer of EMT in melanoma cells. The EMT profile acquired by MSC-CM-exposed melanoma cells is characterized by an enhanced level of mesenchymal markers, including TGFβ/TGFβ-receptors system upregulation, by increased invasiveness and uPAR expression, and in vivo tumor growth. Silencing TGFβ in MSC is found to abrogate ability of MSC to promote EMT characteristics in melanoma cells, together with uPAR expression, and this finding is strengthened using an antagonist peptide of TGFβRIII, the so-called P17. Finally, we demonstrate that the uPAR antisense oligonucleotide (uPAR aODN) may inhibit EMT of melanoma cells either stimulated by exogenous TGFβ or MSC-CM. Thus, uPAR upregulation in melanoma cells exposed to MSC-medium drives TGFβ-mediated EMT. On the whole, TGFβ/uPAR dangerous liaison in cancer cell/MSC interactions may disclose a new strategy to abrogate melanoma progression. Mesenchymal stem cell (MSC)-conditioned medium induces EMT-like profile in melanoma. MSC-derived TGFβ promotes uPAR and TGFβ/TGFβ-receptor upregulation in melanoma. TGFβ gene silencing in MSCs downregulates uPAR expression and EMT in melanoma. uPAR downregulation prevents MSC-induced EMT-like profile in melanoma cells. Inhibition of the dangerous TGFβ/uPAR relationship might abrogate melanoma progression.

  2. Prognosis in adenocarcinomas of lower oesophagus, gastro-oesophageal junction and cardia evaluated by uPAR-immunohistochemistry

    DEFF Research Database (Denmark)

    Laerum, Ole Didrik; Ovrebo, Kjell; Skarstein, Arne

    2012-01-01

    Adenocarcinomas of lower oesophagus, gastro-oesophageal junction and cardia in humans are highly invasive tumours with poor prognosis. The localisation of urokinase-type plasminogen activator receptor (uPAR) was determined in 66 patients; 60 with adenocarcinomas and six cases with Barrett......'s oesophagus. uPAR was expressed in nearly all cases of invasive adenocarcinomas by populations of cancer cells, macrophages and myofibroblasts at both the invasion front and the tumour core. In areas with high-grade dysplasia or with Barrett's metaplasia adjacent to the tumour tissue, no u......PAR-positivity on immmunohistochemically stained specimens, a significant association was found between poor overall survival and high uPAR-score for cancer cells in the tumour core and for macrophages peripherally at the tumour invasion zone. In multivariate analysis, these two uPAR-scores were confirmed as highly significant prognostic...

  3. Tumour microenvironments induce expression of urokinase plasminogen activator receptor (uPAR and concomitant activation of gelatinolytic enzymes.

    Directory of Open Access Journals (Sweden)

    Synnøve Magnussen

    Full Text Available The urokinase plasminogen activator receptor (uPAR is associated with poor prognosis in oral squamous cell carcinoma (OSCC, and increased expression of uPAR is often found at the invasive tumour front. The aim of the current study was to elucidate the role of uPAR in invasion and metastasis of OSCC, and the effects of various tumour microenvironments in these processes. Furthermore, we wanted to study whether the cells' expression level of uPAR affected the activity of gelatinolytic enzymes.The Plaur gene was both overexpressed and knocked-down in the murine OSCC cell line AT84. Tongue and skin tumours were established in syngeneic mice, and cells were also studied in an ex vivo leiomyoma invasion model. Soluble factors derived from leiomyoma tissue, as well as purified extracellular matrix (ECM proteins, were assessed for their ability to affect uPAR expression, glycosylation and cleavage. Activity of gelatinolytic enzymes in the tissues were assessed by in situ zymography.We found that increased levels of uPAR did not induce tumour invasion or metastasis. However, cells expressing low endogenous levels of uPAR in vitro up-regulated uPAR expression both in tongue, skin and leiomyoma tissue. Various ECM proteins had no effect on uPAR expression, while soluble factors originating from the leiomyoma tissue increased both the expression and glycosylation of uPAR, and possibly also affected the proteolytic processing of uPAR. Tumours with high levels of uPAR, as well as cells invading leiomyoma tissue with up-regulated uPAR expression, all displayed enhanced activity of gelatinolytic enzymes.Although high levels of uPAR are not sufficient to induce invasion and metastasis, the activity of gelatinolytic enzymes was increased. Furthermore, several tumour microenvironments have the capacity to induce up-regulation of uPAR expression, and soluble factors in the tumour microenvironment may have an important role in the regulation of posttranslational

  4. Copenhagen uPAR prostate cancer (CuPCa) database

    DEFF Research Database (Denmark)

    Lippert, Solvej; Berg, Kasper D; Høyer-Hansen, Gunilla

    2016-01-01

    AIM: Urokinase plasminogen activator receptor (uPAR) plays a central role during cancer invasion by facilitating pericellular proteolysis. We initiated the prospective 'Copenhagen uPAR Prostate Cancer' study to investigate the significance of uPAR levels in prostate cancer (PCa) patients. METHODS...

  5. Targeting of peptide conjugated magnetic nanoparticles to urokinase plasminogen activator receptor (uPAR) expressing cells

    DEFF Research Database (Denmark)

    Hansen, Line; Unmack Larsen, Esben Kjær; Nielsen, Erik Holm

    2013-01-01

    patient prognosis shared by several cancers including breast, colorectal, and gastric cancers. Conjugation of a uPAR specific targeting peptide onto polyethylene glycol (PEG) coated USPIO nanoparticles by click chemistry resulted in a five times higher uptake in vitro in a uPAR positive cell line compared......Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles are currently being used as a magnetic resonance imaging (MRI) contrast agent in vivo, mainly by their passive accumulation in tissues of interest. However, a higher specificity can ideally be achieved when the nanoparticles are targeted...... towards cell specific receptors and this may also facilitate specific drug delivery by an enhanced target-mediated endocytosis. We report efficient peptide-mediated targeting of magnetic nanoparticles to cells expressing the urokinase plasminogen activator receptor (uPAR), a surface biomarker for poor...

  6. uPAR as anti-cancer target

    DEFF Research Database (Denmark)

    Lund, Ida K; Illemann, Martin; Thurison, Tine

    2011-01-01

    , and a potential diagnostic and predictive impact of the different uPAR forms has been reported. Hence, pericellular proteolysis seems to be a suitable target for anti-cancer therapy and numerous approaches have been pursued. Targeting of this process may be achieved by preventing the binding of uPA to u...... using mouse monoclonal antibodies (mAbs) against mouse uPA or uPAR. These reagents will target uPA and uPAR in both stromal cells and cancer cells, and their therapeutic potential can now be assessed in syngenic mouse cancer models....

  7. First-in-human uPAR PET

    DEFF Research Database (Denmark)

    Persson, Morten; Skovgaard, Dorthe; Brandt-Larsen, Malene

    2015-01-01

    A first-in-human clinical trial with Positron Emission Tomography (PET) imaging of the urokinase-type plasminogen activator receptor (uPAR) in patients with breast, prostate and bladder cancer, is described. uPAR is expressed in many types of human cancers and the expression is predictive...... of invasion, metastasis and indicates poor prognosis. uPAR PET imaging therefore holds promise to be a new and innovative method for improved cancer diagnosis, staging and individual risk stratification. The uPAR specific peptide AE105 was conjugated to the macrocyclic chelator DOTA and labeled with (64)Cu...... for targeted molecular imaging with PET. The safety, pharmacokinetic, biodistribution profile and radiation dosimetry after a single intravenous dose of (64)Cu-DOTA-AE105 were assessed by serial PET and computed tomography (CT) in 4 prostate, 3 breast and 3 bladder cancer patients. Safety assessment...

  8. Peptide-Based Optical uPAR Imaging for Surgery

    DEFF Research Database (Denmark)

    Juhl, Karina; Christensen, Anders; Persson, Morten

    2016-01-01

    Near infrared intra-operative optical imaging is an emerging technique with clear implications for improved cancer surgery by enabling a more distinct delineation of the tumor margins during resection. This modality has the potential to increase the number of patients having a curative radical...... tumor resection. In the present study, a new uPAR-targeted fluorescent probe was developed and the in vivo applicability was evaluated in a human xenograft mouse model. Most human carcinomas express high level of uPAR in the tumor-stromal interface of invasive lesions and uPAR is therefore considered...... an ideal target for intra-operative imaging. Conjugation of the flourophor indocyanine green (ICG) to the uPAR agonist (AE105) provides an optical imaging ligand with sufficiently high receptor affinity to allow for a specific receptor targeting in vivo. For in vivo testing, human glioblastoma xenograft...

  9. The urokinase receptor (uPAR) and the uPAR-associated protein (uPARAP/Endo180)

    DEFF Research Database (Denmark)

    Behrendt, Niels

    2004-01-01

    The breakdown of the barriers formed by extracellular matrix proteins is a pre-requisite for all processes of tissue remodeling. Matrix degradation reactions take part in specific physiological events in the healthy organism but also represent a crucial step in cancer invasion. These degradation...... processes involve a highly organized interplay between proteases and their cellular binding sites as well as specific substrates and internalization receptors. This review article is focused on two components, the urokinase plasminogen activator receptor (uPAR) and the uPAR-associated protein (uPARAP, also...... designated Endo180), that are considered crucially engaged in matrix degradation. uPAR and uPARAP have highly diverse functions, but on certain cell types they interact with each other in a process that is still incompletely understood. uPAR is a glycosyl-phosphatidylinositol-anchored glycoprotein...

  10. Urokinase-type plasminogen activator receptor (uPAR) on tumor-associated macrophages is a marker of poor prognosis in colorectal cancer

    DEFF Research Database (Denmark)

    Illemann, Martin; Laerum, Ole Didrik; Hasselby, Jane Preuss

    2014-01-01

    Patients were identified from a population-based prospective study of 4990 individuals with symptoms associated with colorectal cancer (CRC). A total of 244 CRC tissue samples were available for immunohistochemical staining of uPAR, semiquantitatively scored at the invasive front, and in the tumor.......0009]) and cancer cells (HR=1.49, [95% CI: 1.01-2.20, P = 0.047]) located in the tumor core were significantly associated to overall survival. In a multivariate analysis, uPAR-positive versus uPAR-negative macrophages located in the tumor core showed the best separation of patients with positive score associated...... significantly associated to overall survival (HR = 1.81 [95% CI: 1.08-3.01, P = 0.023]). Membrane-bound uPAR showed additive effects with the circulating uPAR(I) and stage, giving a hazard ratio of 12 between low and high scores. Thus, combining stage, uPAR(I) in blood and uPAR on macrophages in the tumor core...

  11. Intact and cleaved uPAR forms: diagnostic and prognostic value in cancer

    DEFF Research Database (Denmark)

    Rasch, M.G.; Lund, I.K.; Hoyer-Hansen, G.

    2008-01-01

    PA-catalyzed cleavage of uPAR is fast on the cell surface, when uPA is bound to a neighboring uPAR molecule. uPAR can be shed from the cell surface. However, the soluble form cannot be cleaved by uPA. Glycolipid-anchored and soluble forms of intact, uPAR(I-III), and cleaved receptor, uPAR(II-III) and uPAR(I), have been...... identified in tissue and body fluids. It is well-established, that the total amount of all uPAR forms is a strong prognostic marker in different types of cancer. Using immunoassays, measuring the individual uPAR forms, has revealed that the cleaved uPAR forms are even stronger prognostic markers and have...

  12. 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 Masson's trichrome stained equivalent show the viability of the method.

  13. uPAR Targeted Radionuclide Therapy with 177Lu-DOTA-AE105 Inhibits Dissemination of Metastatic Prostate Cancer

    DEFF Research Database (Denmark)

    Persson, Morten; Juhl, Karina; Rasmussen, Palle

    2014-01-01

    activity: a uPAR-targeting probe (177Lu-DOTA-AE105) and a nonbinding control (177Lu-DOTA-AE105mut). Both uPAR flow cytometry and ELISA confirmed high expression levels of the target uPAR in PC-3M-LUC2.luc cells, and cell binding studies using 177Lu-DOTA-AE105 resulted in a specific binding with an IC50....... Moreover, we found a significantly longer metastatic-free survival, with 65% of all mice without any disseminated metastatic lesions present at 65 days after first treatment dose (p = 0.047). In contrast, only 30% of all mice in the combined control groups treated with 177Lu-DOTA-AE105mut or vehicle were...... without metastatic lesions. No treatment-induced toxicity was observed during the study as evaluated by observing animal weight and H&E staining of kidney tissue (dose-limiting organ). Finally, uPAR PET imaging using 64Cu-DOTA-AE105 detected all small, disseminated metastatic foci when compared...

  14. The Urokinase Receptor (uPAR) Facilitates Clearance of Borrelia burgdorferi

    NARCIS (Netherlands)

    Hovius, Joppe W. R.; Bijlsma, Maarten F.; van der Windt, Gerritje J. W.; Wiersinga, W. Joost; Boukens, Bastiaan J. D.; Coumou, Jeroen; Oei, Anneke; de Beer, Regina; de Vos, Alex F.; van 't Veer, Cornelis; van Dam, Alje P.; Wang, Penghua; Fikrig, Erol; Levi, Marcel M.; Roelofs, Joris J. T. H.; van der Poll, Tom

    2009-01-01

    The causative agent of Lyme borreliosis, the spirochete Borrelia burgdorferi, has been shown to induce expression of the urokinase receptor (uPAR); however, the role of uPAR in the immune response against Borrelia has never been investigated. uPAR not only acts as a proteinase receptor, but can

  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. First (18)F-labeled ligand for PET imaging of uPAR

    DEFF Research Database (Denmark)

    Persson, Morten; Liu, Hongguang; Madsen, Jacob

    2013-01-01

    Urokinase-type plasminogen activator receptor (uPAR) is overexpressed in human prostate cancer and uPAR has been found to be associated with metastatic disease and poor prognosis. AE105 is a small linear peptide with high binding affinity to uPAR. We synthesized an N-terminal NOTA-conjugated vers......-conjugated version (NOTA-AE105) for development of the first (18)F-labeled uPAR positron-emission-tomography PET ligand using the Al(18)F radiolabeling method. In this study, the potential of (18)F-AlF-NOTA-AE105 to specifically target uPAR-positive prostate tumors was investigated....

  17. Urokinase-type plasminogen activator receptor (uPAR) as a promising new imaging target

    DEFF Research Database (Denmark)

    Persson, Morten; Kjaer, Andreas

    2013-01-01

    Urokinase-type plasminogen activator receptor (uPAR) has been shown to be of special importance during cancer invasion and metastasis. However, currently, tissue samples are needed for measurement of uPAR expression limiting the potential as a clinical routine. Therefore, non-invasive methods are...... will discuss the potential future clinical application for uPAR imaging as a new imaging biomarker....

  18. The urokinase receptor (uPAR) facilitates clearance of Borrelia burgdorferi.

    OpenAIRE

    Hovius, Joppe W. R; Bijlsma, Maarten F.; van der Windt, Gerritje J.W.; W Joost Wiersinga; Bastiaan J D Boukens; Jeroen Coumou; Anneke Oei; Regina de Beer; de Vos, Alex F; Veer, Cornelis van 't; van Dam, Alje P.; Penghua Wang; Erol Fikrig; Levi, Marcel M; Roelofs, Joris J. T. H.

    2009-01-01

    The causative agent of Lyme borreliosis, the spirochete Borrelia burgdorferi, has been shown to induce expression of the urokinase receptor (uPAR); however, the role of uPAR in the immune response against Borrelia has never been investigated. uPAR not only acts as a proteinase receptor, but can also, dependently or independently of ligation to uPA, directly affect leukocyte function. We here demonstrate that uPAR is upregulated on murine and human leukocytes upon exposure to B. burgdorferi bo...

  19. Multispectral image enhancement for H&E stained pathological tissue specimens

    Science.gov (United States)

    Bautista, Pinky A.; Abe, Tokiya; Yamaguchi, Masahiro; Ohyama, Nagaaki; Yagi, Yukako

    2008-03-01

    The presence of a liver disease such as cirrhosis can be determined by examining the proliferation of collagen fiber from a tissue slide stained with special stain such as the Masson's trichrome(MT) stain. Collagen fiber and smooth muscle, which are both stained the same in an H&E stained slide, are stained blue and pink respectively in an MT-stained slide. In this paper we show that with multispectral imaging the difference between collagen fiber and smooth muscle can be visualized even from an H&E stained image. In the method M KL bases are derived using the spectral data of those H&E stained tissue components which can be easily differentiated from each other, i.e. nucleus, cytoplasm, red blood cells, etc. and based on the spectral residual error of fiber weighting factors are determined to enhance spectral features at certain wavelengths. Results of our experiment demonstrate the capability of multispectral imaging and its advantage compared to the conventional RGB imaging systems to delineate tissue structures with subtle colorimetric difference.

  20. Suppression of uPAR retards radiation-induced invasion and migration mediated by integrin β1/FAK signaling in medulloblastoma.

    Directory of Open Access Journals (Sweden)

    Arun Kumar Nalla

    2010-09-01

    Full Text Available Despite effective radiotherapy for the initial stages of cancer, several studies have reported the recurrence of various cancers, including medulloblastoma. Here, we attempt to capitalize on the radiation-induced aggressive behavior of medulloblastoma cells by comparing the extracellular protease activity and the expression pattern of molecules, known to be involved in cell adhesion, migration and invasion, between non-irradiated and irradiated cells.We identified an increase in invasion and migration of irradiated compared to non-irradiated medulloblastoma cells. RT-PCR analysis confirmed increased expression of uPA, uPAR, focal adhesion kinase (FAK, N-Cadherin and integrin subunits (e.g., α3, α5 and β1 in irradiated cells. Furthermore, we noticed a ∼2-fold increase in tyrosine phosphorylation of FAK in irradiated cells. Immunoprecipitation studies confirmed increased interaction of integrin β1 and FAK in irradiated cells. In addition, our results show that overexpression of uPAR in cancer cells can mimic radiation-induced activation of FAK signaling. Moreover, by inhibiting FAK phosphorylation, we were able to reduce the radiation-induced invasiveness of the cancer cells. In this vein, we studied the effect of siRNA-mediated knockdown of uPAR on cell migration and adhesion in irradiated and non-irradiated medulloblastoma cells. Downregulation of uPAR reduced the radiation-induced adhesion, migration and invasion of the irradiated cells, primarily by inhibiting phosphorylation of FAK, Paxillin and Rac-1/Cdc42. As observed from the immunoprecipitation studies, uPAR knockdown reduced interaction among the focal adhesion molecules, such as FAK, Paxillin and p130Cas, which are known to play key roles in cancer metastasis. Pretreatment with uPAR shRNA expressing construct reduced uPAR and phospho FAK expression levels in pre-established medulloblastoma in nude mice.Taken together, our results show that radiation enhances uPAR-mediated FAK

  1. Digital staining for histopathology multispectral images by the combined application of spectral enhancement and spectral transformation.

    Science.gov (United States)

    Bautista, Pinky A; Yagi, Yukako

    2011-01-01

    In this paper we introduced a digital staining method for histopathology images captured with an n-band multispectral camera. The method consisted of two major processes: enhancement of the original spectral transmittance and the transformation of the enhanced transmittance to its target spectral configuration. Enhancement is accomplished by shifting the original transmittance with the scaled difference between the original transmittance and the transmittance estimated with m dominant principal component (PC) vectors;the m-PC vectors were determined from the transmittance samples of the background image. Transformation of the enhanced transmittance to the target spectral configuration was done using an nxn transformation matrix, which was derived by applying a least square method to the enhanced and target spectral training data samples of the different tissue components. Experimental results on the digital conversion of a hematoxylin and eosin (H&E) stained multispectral image to its Masson's trichrome stained (MT) equivalent shows the viability of the method.

  2. 177-lu labeled peptide for site-specific uPAR-targeting

    DEFF Research Database (Denmark)

    2015-01-01

    There is provided a 177-Lu labelled peptide for site-specific targeting of the Urokinase Plasminogen Activator Receptor (uPAR) thereby enabling treatment of a cancer disease associated with high uPAR expression; e.g. treatment of colorectal cancer by administering to a patient an effective amount...... of the 177-Lu labelled peptide....

  3. Enhancement of seminal stains using background correction algorithm with colour filters.

    Science.gov (United States)

    Lee, Wee Chuen; Khoo, Bee Ee; Abdullah, Ahmad Fahmi Lim

    2016-06-01

    Evidence in crime scenes available in the form of biological stains which cannot be visualized during naked eye examination can be detected by imaging their fluorescence using a combination of excitation lights and suitable filters. These combinations selectively allow the passage of fluorescence light emitted from the targeted stains. However, interference from the fluorescence generated by many of the surface materials bearing the stains often renders it difficult to visualize the stains during forensic photography. This report describes the use of background correction algorithm (BCA) to enhance the visibility of seminal stain, a biological evidence that fluoresces. While earlier reports described the use of narrow band-pass filters for other fluorescing evidences, here, we utilize BCA to enhance images captured using commonly available colour filters, yellow, orange and red. Mean-based contrast adjustment was incorporated into BCA to adjust the background brightness for achieving similarity of images' background appearance, a crucial step for ensuring success while implementing BCA. Experiment results demonstrated the effectiveness of our proposed colour filters' approach using the improved BCA in enhancing the visibility of seminal stains in varying dilutions on selected surfaces. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  4. Color Swapping to Enhance Breast Cancer Digital Images Qualities Using Stain Normalization

    Science.gov (United States)

    Muhimmah, Izzati; Puspasari Wijaya, Dhina; Indrayanti

    2017-03-01

    Histopathology is the disease diagnosis by means of the visual examination of tissues under the microscope. The virtually transparent tissue sections were prepared using a number of colored histochemical stains bound selectively to the cellular components. A variation of colors comes to be a problem in histopathology based upon the microscope lighting for the range of factors. This research aimed to investigate an image enhancement by applying a nonlinear mapping approach to stain normalization and histogram equalization for contrast enhancement. Validation was carried out in 59 datasets with 96.6% accordance and expert justification.

  5. Identification of uPAR-positive chemoresistant cells in small cell lung cancer.

    Directory of Open Access Journals (Sweden)

    Margarita Gutova

    Full Text Available BACKGROUND: The urokinase plasminogen activator (uPA and its receptor (uPAR/CD87 are major regulators of extracellular matrix degradation and are involved in cell migration and invasion under physiological and pathological conditions. The uPA/uPAR system has been of great interest in cancer research because it is involved in the development of most invasive cancer phenotypes and is a strong predictor of poor patient survival. However, little is known about the role of uPA/uPAR in small cell lung cancer (SCLC, the most aggressive type of lung cancer. We therefore determined whether uPA and uPAR are involved in generation of drug resistant SCLC cell phenotype. METHODS AND FINDINGS: We screened six human SCLC cell lines for surface markers for putative stem and cancer cells. We used fluorescence-activated cell sorting (FACS, fluorescence microscopy and clonogenic assays to demonstrate uPAR expression in a subpopulation of cells derived from primary and metastatic SCLC cell lines. Cytotoxic assays were used to determine the sensitivity of uPAR-positive and uPAR-negative cells to chemotherapeutic agents. The uPAR-positive cells in all SCLC lines demonstrated multi-drug resistance, high clonogenic activity and co-expression of CD44 and MDR1, putative cancer stem cell markers. CONCLUSIONS: These data suggest that uPAR-positive cells may define a functionally important population of cancer cells in SCLC, which are resistant to traditional chemotherapies, and could serve as critical targets for more effective therapeutic interventions in SCLC.

  6. The urokinase receptor (uPAR facilitates clearance of Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Joppe W R Hovius

    2009-05-01

    Full Text Available The causative agent of Lyme borreliosis, the spirochete Borrelia burgdorferi, has been shown to induce expression of the urokinase receptor (uPAR; however, the role of uPAR in the immune response against Borrelia has never been investigated. uPAR not only acts as a proteinase receptor, but can also, dependently or independently of ligation to uPA, directly affect leukocyte function. We here demonstrate that uPAR is upregulated on murine and human leukocytes upon exposure to B. burgdorferi both in vitro as well as in vivo. Notably, B. burgdorferi-inoculated C57BL/6 uPAR knock-out mice harbored significantly higher Borrelia numbers compared to WT controls. This was associated with impaired phagocytotic capacity of B. burgdorferi by uPAR knock-out leukocytes in vitro. B. burgdorferi numbers in vivo, and phagocytotic capacity in vitro, were unaltered in uPA, tPA (low fibrinolytic activity and PAI-1 (high fibrinolytic activity knock-out mice compared to WT controls. Strikingly, in uPAR knock-out mice partially backcrossed to a B. burgdorferi susceptible C3H/HeN background, higher B. burgdorferi numbers were associated with more severe carditis and increased local TLR2 and IL-1beta mRNA expression. In conclusion, in B. burgdorferi infection, uPAR is required for phagocytosis and adequate eradication of the spirochete from the heart by a mechanism that is independent of binding of uPAR to uPA or its role in the fibrinolytic system.

  7. The Urokinase Receptor (uPAR) Facilitates Clearance of Borrelia burgdorferi

    Science.gov (United States)

    Hovius, Joppe W. R.; Bijlsma, Maarten F.; van der Windt, Gerritje J. W.; Wiersinga, W. Joost; Boukens, Bastiaan J. D.; Coumou, Jeroen; Oei, Anneke; de Beer, Regina; de Vos, Alex F.; Veer, Cornelis van 't; van Dam, Alje P.; Wang, Penghua; Fikrig, Erol; Levi, Marcel M.; Roelofs, Joris J. T. H.; van der Poll, Tom

    2009-01-01

    The causative agent of Lyme borreliosis, the spirochete Borrelia burgdorferi, has been shown to induce expression of the urokinase receptor (uPAR); however, the role of uPAR in the immune response against Borrelia has never been investigated. uPAR not only acts as a proteinase receptor, but can also, dependently or independently of ligation to uPA, directly affect leukocyte function. We here demonstrate that uPAR is upregulated on murine and human leukocytes upon exposure to B. burgdorferi both in vitro as well as in vivo. Notably, B. burgdorferi-inoculated C57BL/6 uPAR knock-out mice harbored significantly higher Borrelia numbers compared to WT controls. This was associated with impaired phagocytotic capacity of B. burgdorferi by uPAR knock-out leukocytes in vitro. B. burgdorferi numbers in vivo, and phagocytotic capacity in vitro, were unaltered in uPA, tPA (low fibrinolytic activity) and PAI-1 (high fibrinolytic activity) knock-out mice compared to WT controls. Strikingly, in uPAR knock-out mice partially backcrossed to a B. burgdorferi susceptible C3H/HeN background, higher B. burgdorferi numbers were associated with more severe carditis and increased local TLR2 and IL-1β mRNA expression. In conclusion, in B. burgdorferi infection, uPAR is required for phagocytosis and adequate eradication of the spirochete from the heart by a mechanism that is independent of binding of uPAR to uPA or its role in the fibrinolytic system. PMID:19461880

  8. Data on the regulation of moesin and merlin by the urokinase receptor (uPAR): Model explaining distal activation of integrins by uPAR.

    Science.gov (United States)

    Degryse, Bernard; Britto, Mishan; Shan, Chun Xu; Wallace, Robert G; Rochfort, Keith D; Cummins, Philip M; Meade, Gerardene; Murphy, Ronan P

    2017-12-01

    The data presented herein are connected to our research article (doi: 10.1016/j.biocel.2017.04.012) [1], in which we investigated the functional connections between the urokinase receptor (uPAR), and the ezrin/radixin/moesin (ERM) proteins, moesin and merlin [1]. Firstly, a model of action is proposed that enlightens how uPAR regulates distal integrins. In addition, data show the effects of expressing wild-type moesin or permanently active T558D mutant of moesin on angiogenesis and morphology of human aortic endothelial cells (HAEC). Additional data compare the effects of urokinase (uPA, the main ligand of uPAR) on the same cells. Lastly, we provide technical data demonstrating the effects of specific siRNA for moesin and merlin on moesin and merlin expression, respectively.

  9. Data on the regulation of moesin and merlin by the urokinase receptor (uPAR: Model explaining distal activation of integrins by uPAR

    Directory of Open Access Journals (Sweden)

    Bernard Degryse

    2017-12-01

    Full Text Available The data presented herein are connected to our research article (doi: 10.1016/j.biocel.2017.04.012 [1], in which we investigated the functional connections between the urokinase receptor (uPAR, and the ezrin/radixin/moesin (ERM proteins, moesin and merlin [1]. Firstly, a model of action is proposed that enlightens how uPAR regulates distal integrins. In addition, data show the effects of expressing wild-type moesin or permanently active T558D mutant of moesin on angiogenesis and morphology of human aortic endothelial cells (HAEC. Additional data compare the effects of urokinase (uPA, the main ligand of uPAR on the same cells. Lastly, we provide technical data demonstrating the effects of specific siRNA for moesin and merlin on moesin and merlin expression, respectively. Keywords: Urokinase receptor, Moesin, Merlin, Angiogenesis, siRNA

  10. uPAR Expression Pattern in Patients with Urothelial Carcinoma of the Bladder

    DEFF Research Database (Denmark)

    Dohn, Line Hammer; Pappot, Helle; Iversen, Benedikte Richter

    2015-01-01

    The objective of the present study was to confirm the expression and localisation pattern of the urokinase-type plasminogen activator receptor (uPAR) focusing on its possible clinical relevance in patients with urothelial neoplasia of the bladder. uPAR is a central molecule in tissue remodelling...... during cancer invasion and metastasis and is an established prognostic marker in various cancer diseases other than bladder cancer. Formalin-fixed and paraffin-embedded tumour-tissue blocks from 186 patients treated with radical cystectomy were analysed. uPAR expression was scored as either negative...

  11. The superoxide scavenger TEMPOL induces urokinase receptor (uPAR expression in human prostate cancer cells

    Directory of Open Access Journals (Sweden)

    Francis Joseph

    2006-06-01

    Full Text Available Abstract There is little understanding of the effect that reactive oxygen metabolites have on cellular behavior during the processes of invasion and metastasis. These oxygen metabolites could interact with a number of targets modulating their function such as enzymes involved in basement membrane dissolution, adhesion molecules involved in motility or receptors involved in proliferation. We investigated the effect of increased scavenging of superoxide anions on the expression of the urokinase receptor (uPAR in PC-3M human prostate cancer cells. Urokinase receptor is a GPI-linked cell surface molecule which mediates multiple functions including adhesion, proliferation and pericellular proteolysis. Addition of the superoxide scavenger 4-hydroxy-2,2,6,6-tetramethylpiperidinyloxy (TEMPOL to PC-3M cultures stimulated expression of uPAR protein peaking between 48 and 72 hours. Cell surface expression of the uPAR was also increased. Surprisingly, uPAR transcript levels increased only slightly and this mild increase did not coincide with the striking degree of protein increase. This disparity indicates that the TEMPOL effect on uPAR occurs through a post-transcriptional mechanism. TEMPOL presence in PC-3M cultures reduced intracellular superoxide-type species by 75% as assayed by NBT dye conversion; however this reduction significantly diminished within hours following TEMPOL removal. The time gap between TEMPOL treatment and peak uPAR protein expression suggests that reduction of reactive oxygen metabolites in prostate cancer cells initiates a multistep pathway which requires several hours to culminate in uPAR induction. These findings reveal a novel pathway for uPAR regulation involving reactive oxygens such as superoxide anion.

  12. Chemical enhancement of footwear impressions in blood on fabric - part 3: amino acid staining.

    Science.gov (United States)

    Farrugia, Kevin J; Bandey, Helen; Savage, Kathleen; NicDaéid, Niamh

    2013-03-01

    Enhancement of footwear impressions, using ninhydrin or ninhydrin analogues is not considered common practice and such techniques are generally used to target amino acids present in fingermarks where the reaction gives rise to colour and possibly fluorescence. Ninhydrin and two of its analogues were used for the enhancement of footwear impressions in blood on various types, colours and porosities of fabric. Test footwear impressions on fabric were prepared using a specifically built rig to minimise the variability between each impression. Ninhydrin enhancement of footwear impressions in blood on light coloured fabric yielded good enhancement results, however the contrast was weak or non-existent on dark coloured fabrics. Other ninhydrin analogues which have the advantage of fluorescence failed to enhance the impressions in blood on all fabrics. The sequential treatment of impressions in blood on fabric with other blood enhancing reagents (e.g. protein stains and heme reagents) was also investigated. Copyright © 2012 Forensic Science Society. All rights reserved.

  13. uPA/uPAR system activation drives a glycolytic phenotype in melanoma cells.

    Science.gov (United States)

    Laurenzana, Anna; Chillà, Anastasia; Luciani, Cristina; Peppicelli, Silvia; Biagioni, Alessio; Bianchini, Francesca; Tenedini, Elena; Torre, Eugenio; Mocali, Alessandra; Calorini, Lido; Margheri, Francesca; Fibbi, Gabriella; Del Rosso, Mario

    2017-09-15

    In this manuscript, we show the involvement of the uPA/uPAR system in the regulation of aerobic glycolysis of melanoma cells. uPAR over-expression in human melanoma cells controls an invasive and glycolytic phenotype in normoxic conditions. uPAR down-regulation by siRNA or its uncoupling from integrins, and hence from integrin-linked tyrosine kinase receptors (IL-TKRs), by an antagonist peptide induced a striking inhibition of the PI3K/AKT/mTOR/HIF1α pathway, resulting into impairment of glucose uptake, decrease of several glycolytic enzymes and of PKM2, a checkpoint that controls metabolism of cancer cells. Further, binding of uPA to uPAR regulates expression of molecules that govern cell invasion, including extracellular matrix metallo-proteinases inducer (EMPPRIN) and enolase, a glycolytyc enzyme that also serves as a plasminogen receptor, thus providing a common denominator between tumor metabolism and phenotypic invasive features. Such effects depend on the α5β1-integrin-mediated uPAR connection with EGFR in melanoma cells with engagement of the PI3K-mTOR-HIFα pathway. HIF-1α trans-activates genes whose products mediate tumor invasion and glycolysis, thus providing the common denominator between melanoma metabolism and its invasive features. These findings unveil a unrecognized interaction between the invasion-related uPAR and IL-TKRs in the control of glycolysis and disclose a new pharmacological target (i.e., uPAR/IL-TKRs axis) for the therapy of melanoma. © 2017 UICC.

  14. Clathrin and LRP-1-independent constitutive endocytosis and recycling of uPAR.

    Directory of Open Access Journals (Sweden)

    Katia Cortese

    Full Text Available BACKGROUND: The urokinase receptor (uPAR/CD87 is highly expressed in malignant tumours. uPAR, as a GPI anchored protein, is preferentially located at the cell surface, where it interacts with its ligands urokinase (uPA and the extracellular matrix protein vitronectin, thus promoting plasmin generation, cell-matrix interactions and intracellular signalling events. Interaction with a complex formed by uPA and its inhibitor PAI-1 induces cell surface down regulation and recycling of the receptor via the clathrin-coated pathway, a process dependent on the association to LRP-1. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we have found that along with the ligand-induced down-regulation, uPAR also internalizes and recycles constitutively through a second pathway that is independent of LRP-1 and clathrin but shares some properties with macropinocytosis. The ligand-independent route is amiloride-sensitive, does not require uPAR partitioning into lipid rafts, is independent of the activity of small GTPases RhoA, Rac1 and Cdc42, and does not require PI3K activity. Constitutively endocytosed uPAR is found in EEA1 positive early/recycling endosomes but does not reach lysosomes in the absence of ligands. Electron microscopy analysis reveals the presence of uPAR in ruffling domains at the cell surface, in macropinosome-like vesicles and in endosomal compartments. CONCLUSIONS/SIGNIFICANCE: These results indicate that, in addition to the ligand-induced endocytosis of uPAR, efficient surface expression and membrane trafficking might also be driven by an uncommon macropinocytic mechanism coupled with rapid recycling to the cell surface.

  15. Enhanced Port Wine Stain Lightening Achieved with Combined Treatment of Selective Photothermolysis and Imiquimod

    Science.gov (United States)

    Tremaine, Anne Marie; Armstrong, Jennifer; Huang, Yu-Chih; Elkeeb, Leila; Ortiz, Arisa; Harris, Ronald; Choi, Bernard; Kelly, Kristen M.

    2012-01-01

    Background Pulsed dye laser is the gold standard for treatment of port wine stain birthmarks but multiple treatments are required and complete resolution is often not achieved. Post-treatment vessel recurrence is thought to be a factor that limits efficacy of pulsed dye laser treatment of port wine stains. Imiquimod 5% cream is an immunomodulator with anti-angiogenic effects. Objective To determine if application of imiquimod 5% cream after pulsed dye laser improves treatment outcome. Methods Healthy patients with port wine stains (n = 24) were treated with pulsed dye laser and then randomized to apply post-treatment placebo or imiquimod 5% cream for 8 weeks. Chromameter measurements (CIE L*a*b* colorspace) for 57 port wine stain sites (multiple sites per subject) were taken at baseline and compared with measurements taken 8 weeks post-treatment. The change in a* and ΔE were measured to quantify treatment outcome. Results Two subjects developed minor skin irritation. Other adverse effects weren't noted. Average Δa* was 0.43 for pulsed dye laser + placebo sites (n = 25) and 1.27 for pulsed dye laser + imiquimod sites (n = 32) (p value = 0.0294) indicating a greater reduction in erythema with imiquimod. Average ΔE was 2.59 for pulsed dye laser + placebo and 4.08 for pulsed dye laser + imiquimod (p value = 0.0363), again indicating a greater color improvement with imiquimod. Limitations Effects were evaluated after a single treatment and duration of effect is unknown. Conclusion Combined selective photothermolysis and anti-angiogenic therapy may enhance port wine stain treatment efficacy. PMID:22244840

  16. Identification of a new epitope in uPAR as a target for the cancer therapeutic monoclonal antibody ATN-658, a structural homolog of the uPAR binding integrin CD11b (αM.

    Directory of Open Access Journals (Sweden)

    Xiang Xu

    Full Text Available The urokinase plasminogen activator receptor (uPAR plays a role in tumor progression and has been proposed as a target for the treatment of cancer. We recently described the development of a novel humanized monoclonal antibody that targets uPAR and has anti-tumor activity in multiple xenograft animal tumor models. This antibody, ATN-658, does not inhibit ligand binding (i.e. uPA and vitronectin to uPAR and its mechanism of action remains unclear. As a first step in understanding the anti-tumor activity of ATN-658, we set out to identify the epitope on uPAR to which ATN-658 binds. Guided by comparisons between primate and human uPAR, epitope mapping studies were performed using several orthogonal techniques. Systematic site directed and alanine scanning mutagenesis identified the region of aa 268-275 of uPAR as the epitope for ATN-658. No known function has previously been attributed to this epitope Structural insights into epitope recognition were obtained from structural studies of the Fab fragment of ATN-658 bound to uPAR. The structure shows that the ATN-658 binds to the DIII domain of uPAR, close to the C-terminus of the receptor, corroborating the epitope mapping results. Intriguingly, when bound to uPAR, the complementarity determining region (CDR regions of ATN-658 closely mimic the binding regions of the integrin CD11b (αM, a previously identified uPAR ligand thought to be involved in leukocyte rolling, migration and complement fixation with no known role in tumor progression of solid tumors. These studies reveal a new functional epitope on uPAR involved in tumor progression and demonstrate a previously unrecognized strategy for the therapeutic targeting of uPAR.

  17. PET imaging of urokinase-type plasminogen activator receptor (uPAR) in prostate cancer

    DEFF Research Database (Denmark)

    Skovgaard, Dorthe; Persson, Morten; Kjaer, Andreas

    2016-01-01

    (intact/cleaved forms)—provides independent additional clinical information to that contributed by PSA, Gleason score, and other relevant pathological and clinical parameters. In this respect, non-invasive molecular imaging by positron emission tomography (PET) offers a very attractive technology platform......, which can provide the required quantitative information on the uPAR expression profile, without the need for invasive procedures and the risk of missing the target due to tumor heterogeneity. These observations support non-invasive PET imaging of uPAR in PC as a clinically relevant diagnostic...

  18. Endothelial progenitor cell-dependent angiogenesis requires localization of the full-length form of uPAR in caveolae.

    Science.gov (United States)

    Margheri, Francesca; Chillà, Anastasia; Laurenzana, Anna; Serratì, Simona; Mazzanti, Benedetta; Saccardi, Riccardo; Santosuosso, Michela; Danza, Giovanna; Sturli, Niccolò; Rosati, Fabiana; Magnelli, Lucia; Papucci, Laura; Calorini, Lido; Bianchini, Francesca; Del Rosso, Mario; Fibbi, Gabriella

    2011-09-29

    Endothelial urokinase-type plasminogen activator receptor (uPAR) is thought to provide a regulatory mechanism in angiogenesis. Here we studied the proangiogenic role of uPAR in endothelial colony-forming cells (ECFCs), a cell population identified in human umbilical blood that embodies all of the properties of an endothelial progenitor cell matched with a high proliferative rate. By using caveolae-disrupting agents and by caveolin-1 silencing, we have shown that the angiogenic properties of ECFCs depend on caveolae integrity and on the presence of full-length uPAR in such specialized membrane invaginations. Inhibition of uPAR expression by antisense oligonucleotides promoted caveolae disruption, suggesting that uPAR is an inducer of caveolae organization. Vascular endothelial growth factor (VEGF) promoted accumulation of uPAR in ECFC caveolae in its undegraded form. We also demonstrated that VEGF-dependent ERK phosphorylation required integrity of caveolae as well as caveolar uPAR expression. VEGF activity depends on inhibition of ECFC MMP12 production, which results in impairment of MMP12-dependent uPAR truncation. Further, MMP12 overexpression in ECFC inhibited vascularization in vitro and in vivo. Our data suggest that intratumor homing of ECFCs suitably engineered to overexpress MMP12 could have the chance to control uPAR-dependent activities required for tumor angiogenesis and malignant cells spreading.

  19. uPAR-targeted multimodal tracer for pre- and intraoperative imaging in cancer surgery

    NARCIS (Netherlands)

    Boonstra, M.C.; Van Driel, P.B.A.A.; Van Willigen, D.M.; Stammes, M.A.; Prevoo, H.A.J.M.; Tummers, Q.R.J.G.; Mazar, A.P.; Beekman, F.J.; Kuppen, P.J.K.; Van de Velde, C.J.H.; Löwik, C.W.G.M.; Frangioni, J.V.; Van Leeuwen, F.W.B.; Sier, C.F.M.; Vahrmeijer, A.L.

    2015-01-01

    Pre- and intraoperative diagnostic techniques facilitating tumor staging are of paramount importance in colorectal cancer surgery. The urokinase receptor (uPAR) plays an important role in the development of cancer, tumor invasion, angiogenesis, and metastasis and over-expression is found in the

  20. The interaction between uPAR and vitronectin triggers ligand-independent adhesion signalling by integrins

    Science.gov (United States)

    Ferraris, Gian Maria Sarra; Schulte, Carsten; Buttiglione, Valentina; De Lorenzi, Valentina; Piontini, Andrea; Galluzzi, Massimiliano; Podestà, Alessandro; Madsen, Chris D; Sidenius, Nicolai

    2014-01-01

    The urokinase-type plasminogen activator receptor (uPAR) is a non-integrin vitronectin (VN) cell adhesion receptor linked to the plasma membrane by a glycolipid anchor. Through structure–function analyses of uPAR, VN and integrins, we document that uPAR-mediated cell adhesion to VN triggers a novel type of integrin signalling that is independent of integrin–matrix engagement. The signalling is fully active on VN mutants deficient in integrin binding site and is also efficiently transduced by integrins deficient in ligand binding. Although integrin ligation is dispensable, signalling is crucially dependent upon an active conformation of the integrin and its association with intracellular adaptors such as talin. This non-canonical integrin signalling is not restricted to uPAR as it poses no structural constraints to the receptor mediating cell attachment. In contrast to canonical integrin signalling, where integrins form direct mechanical links between the ECM and the cytoskeleton, the molecular mechanism enabling the crosstalk between non-integrin adhesion receptors and integrins is dependent upon membrane tension. This suggests that for this type of signalling, the membrane represents a critical component of the molecular clutch. PMID:25168639

  1. Structure and ligand interactions of the urokinase receptor (uPAR)

    DEFF Research Database (Denmark)

    Kjaergaard, Magnus; Hansen, Line V.; Jacobsen, Benedikte

    2008-01-01

    The urokinase-type plasminogen activator receptor (uPAR or CD87) is a glycolipid-anchored membrane glycoprotein, which is responsible for focalizing plasminogen activation to the cell surface through its high-affinity binding to the serine protease uPA. This tight interaction (KD less than 1 nM) ...

  2. Rational targeting of the urokinase receptor (uPAR): development of antagonists and non-invasive imaging probes

    DEFF Research Database (Denmark)

    Kriegbaum, M C; Persson, M; Haldager, L

    2011-01-01

    -ray crystallography and surface plasmon resonance studies. Importantly, these structural studies also define possible druggable target sites in uPAR for small molecules and provide guidelines for the development of reporter groups applicable for non-invasive molecular imaging of uPAR expression in vivo by positron...

  3. Multispectral Enhancement Method to Increase the Visual Differences of Tissue Structures in Stained Histopathology Images

    Directory of Open Access Journals (Sweden)

    Pinky A. Bautista

    2012-01-01

    Full Text Available In this paper we proposed a multispectral enhancement scheme in which the spectral colors of the stained tissue-structure of interest and its background can be independently modified by the user to further improve their visualization and color discrimination. The colors of the background objects are modified by transforming their N-band spectra through an NxN transformation matrix, which is derived by mapping the representative samples of their original spectra to the spectra of their target colors using least mean square method. On the other hand, the color of the tissue structure of interest is modified by modulating the transformed spectra with the sum of the pixel’s spectral residual-errors at specific bands weighted through an NxN weighting matrix; the spectral error is derived by taking the difference between the pixel’s original spectrum and its reconstructed spectrum using the first M dominant principal component vectors in principal component analysis. Promising results were obtained on the visualization of the collagen fiber and the non-collagen tissue structures, e.g., nuclei, cytoplasm and red blood cells (RBC, in a hematoxylin and eosin (H&E stained image.

  4. Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering.

    Science.gov (United States)

    Xu, Shuping; Ji, Xiaohui; Xu, Weiqing; Li, Xiaoling; Wang, Lianying; Bai, Yubai; Zhao, Bing; Ozaki, Yukihiro

    2004-01-01

    This paper reports a novel immunoassay based on surface-enhanced Raman scattering (SERS) and immunogold labelling with silver staining enhancement. Immunoreactions between immunogold colloids modified by a Raman-active probe molecule (e.g., 4-mercaptobenzoic acid) and antigens, which were captured by antibody-assembled chips such as silicon or quartz, were detected via SERS signals of Raman-active probe molecule. All the self-assembled steps were subjected to the measurements of ultraviolet-visible (UV-vis) spectra to monitor the formation of a sandwich structure onto a substrate. The immunoassay was performed by a sandwich structure consisting of three layers. The first layer was composed of immobilized antibody molecules of mouse polyclonal antibody against Hepatitis B virus surface antigen (PAb) on a silicon or quartz substrate. The second layer was the complementary Hepatitis B virus surface antigen (Antigen) molecules captured by PAb on the substrate. The third layer was composed of the probe-labelling immunogold nanoparticles, which were modified by mouse monoclonal antibody against Hepatitis B virus surface antigen (MAb) and 4-mercaptobenzoic acid (MBA) as the Raman-active probe on the surface of gold colloids. After silver staining enhancement, the antigen is identified by a SERS spectrum of MBA. A working curve of the intensity of a SERS signal at 1585 cm(-1) due to the [small nu](8a) aromatic ring vibration of MBA versus the concentration of analyte (Antigen) was obtained and the non-optimized detection limit for the Hepatitis B virus surface antigen was found to be as low as 0.5 [micro sign]g mL(-1).

  5. uPAR-targeted multimodal tracer for pre- and intraoperative imaging in cancer surgery.

    Science.gov (United States)

    Boonstra, Martin C; van Driel, Pieter B A A; van Willigen, Danny M; Stammes, Marieke A; Prevoo, Hendrica A J M; Tummers, Quirijn R J G; Mazar, Andrew P; Beekman, Freek J; Kuppen, Peter J K; van de Velde, Cornelis J H; Löwik, Clemens W G M; Frangioni, John V; van Leeuwen, Fijs W B; Sier, Cornelis F M; Vahrmeijer, Alexander L

    2015-06-10

    Pre- and intraoperative diagnostic techniques facilitating tumor staging are of paramount importance in colorectal cancer surgery. The urokinase receptor (uPAR) plays an important role in the development of cancer, tumor invasion, angiogenesis, and metastasis and over-expression is found in the majority of carcinomas. This study aims to develop the first clinically relevant anti-uPAR antibody-based imaging agent that combines nuclear (111In) and real-time near-infrared (NIR) fluorescent imaging (ZW800-1). Conjugation and binding capacities were investigated and validated in vitro using spectrophotometry and cell-based assays. In vivo, three human colorectal xenograft models were used including an orthotopic peritoneal carcinomatosis model to image small tumors. Nuclear and NIR fluorescent signals showed clear tumor delineation between 24h and 72h post-injection, with highest tumor-to-background ratios of 5.0 ± 1.3 at 72h using fluorescence and 4.2 ± 0.1 at 24h with radioactivity. 1-2 mm sized tumors could be clearly recognized by their fluorescent rim. This study showed the feasibility of an uPAR-recognizing multimodal agent to visualize tumors during image-guided resections using NIR fluorescence, whereas its nuclear component assisted in the pre-operative non-invasive recognition of tumors using SPECT imaging. This strategy can assist in surgical planning and subsequent precision surgery to reduce the number of incomplete resections.

  6. 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%].

  7. A flexible multidomain structure drives the function of the urokinase-type plasminogen activator receptor (uPAR)

    DEFF Research Database (Denmark)

    Mertens, Haydyn D.T.; Kjærgaard, Magnus; Mysling, Simon

    2012-01-01

    -deuterium exchange, and surface plasmon resonance to develop a structural model describing the allosteric regulation of uPAR. We show that the flexibility of its N-terminal domain provides the key for understanding this allosteric mechanism. Importantly, our model has direct implications for understanding uPAR-assisted...... cell adhesion and migration as well as for translational research including targeted intervention therapy and non-invasive tumor imaging in vivo....

  8. Enhanced port-wine stain lightening achieved with combined treatment of selective photothermolysis and imiquimod.

    Science.gov (United States)

    Tremaine, Anne Marie; Armstrong, Jennifer; Huang, Yu-Chih; Elkeeb, Leila; Ortiz, Arisa; Harris, Ronald; Choi, Bernard; Kelly, Kristen M

    2012-04-01

    Pulsed dye laser (PDL) is the gold standard for treatment of port-wine stain (PWS) birthmarks but multiple treatments are required and complete resolution is often not achieved. Posttreatment vessel recurrence is thought to be a factor that limits efficacy of PDL treatment of PWS. Imiquimod 5% cream is an immunomodulator with antiangiogenic effects. We sought to determine if application of imiquimod 5% cream after PDL improves treatment outcome. Healthy individuals with PWS (n = 24) were treated with PDL and then randomized to apply posttreatment placebo or imiquimod 5% cream for 8 weeks. Chromameter measurements (Commission Internationale de l'Eclairage L∗a∗b∗ colorspace) for 57 PWS sites (multiple sites per patient) were taken at baseline and compared with measurements taken 8 weeks posttreatment. The Δa∗ (change in erythema) and ΔE (difference in color between normal-appearing skin and PWS skin) were measured to quantify treatment outcome. Two patients developed minor skin irritation. Other adverse effects were not noted. Average ∆a∗ was 0.43 for PDL + placebo sites (n = 25) and 1.27 for PDL + imiquimod sites (n = 32) (P value = .0294) indicating a greater reduction in erythema with imiquimod. Average ∆E was 2.59 for PDL + placebo and 4.08 for PDL + imiquimod (P value = .0363), again indicating a greater color improvement with imiquimod. Effects were evaluated after a single treatment and duration of effect is unknown. Combined selective photothermolysis and antiangiogenic therapy may enhance PWS treatment efficacy. Copyright © 2011 American Academy of Dermatology, Inc. Published by Mosby, Inc. All rights reserved.

  9. Structure, function and expression on blood and bone marrow cells of the urokinase-type plasminogen activator receptor, uPAR

    DEFF Research Database (Denmark)

    Plesner, T; Behrendt, N; Ploug, M

    1997-01-01

    ), most importantly PAI-1 and PAI-2, providing means of temporally restricting the process of plasminogen activation. In addition to its role in plasminogen activation, compelling evidence has demonstrated a role for uPAR in cell-cell and cell-extracellular matrix adhesion, both directly and indirectly. u......PAR is directly involved in binding to the extracellular matrix molecule, vitronectin, and the affinity of this binding is increased when uPAR is occupied by (pro-)uPA. A more indirect but presumably very important role of uPAR in cell adhesion seems to be mediated through interactions between uPAR and beta 1......-type plasminogen activator (uPAR) is expressed as a differentiation antigen on cells of the myelomonocytic lineage and as an activation antigen on monocytes and T lymphocytes. Neutrophils contain intracellular reservoirs of uPAR that are translocated to the plasma membrane upon activation, and neutrophils from...

  10. HER2 and uPAR cooperativity contribute to metastatic phenotype of HER2-positive breast cancer

    Science.gov (United States)

    Chandran, Vineesh Indira; Eppenberger-Castori, Serenella; Venkatesh, Thejaswini; Vine, Kara Lea; Ranson, Marie

    2015-01-01

    Human epidermal growth factor receptor type 2 (HER2)-positive breast carcinoma is highly aggressive and mostly metastatic in nature though curable/manageable in part by molecular targeted therapy. Recent evidence suggests a subtype of cells within HER2-positive breast tumors that concomitantly expresses the urokinase plasminogen activator receptor (uPAR) with inherent stem cell/mesenchymal-like properties promoting tumor cell motility and a metastatic phenotype. This HER-positive/uPAR-positive subtype may be partially responsible for the failure of HER2-targeted treatment strategies. Herein we discuss and substantiate the cumulative preclinical and clinical evidence on HER2-uPAR cooperativity in terms of gene co-amplification and/or mRNA/protein co-overexpression. We then propose a regulatory signaling model that we hypothesize to maintain upregulation and cooperativity between HER2 and uPAR in aggressive breast cancer. An improved understanding of the HER2/uPAR interaction in breast cancer will provide critical biomolecular information that may help better predict disease course and response to therapy. PMID:25897424

  11. Enhanced port-wine stain lightening achieved with combined treatment of selective photothermolysis and imiquimod

    OpenAIRE

    Tremaine, AM; Armstrong, J; Huang, YC; Elkeeb, L; Ortiz, A; Harris, R.; Choi, B.; Kelly, KM

    2012-01-01

    Background: Pulsed dye laser (PDL) is the gold standard for treatment of port-wine stain (PWS) birthmarks but multiple treatments are required and complete resolution is often not achieved. Posttreatment vessel recurrence is thought to be a factor that limits efficacy of PDL treatment of PWS. Imiquimod 5% cream is an immunomodulator with antiangiogenic effects. Objective: We sought to determine if application of imiquimod 5% cream after PDL improves treatment outcome. Methods: Healthy individ...

  12. uPAR as anti-cancer target: evaluation of biomarker potential, histological localization, and antibody-based therapy

    DEFF Research Database (Denmark)

    Lund, Ida K; Illemann, Martin; Sørensen, Tine Thurison

    2011-01-01

    Degradation of proteins in the extracellular matrix is crucial for the multistep process of cancer invasion and metastasis. Compelling evidence has demonstrated the urokinase receptor (uPAR) and its cognate ligand, the urokinase plasminogen activator (uPA), to play critical roles in the concerted......, and a potential diagnostic and predictive impact of the different uPAR forms has been reported. Hence, pericellular proteolysis seems to be a suitable target for anti-cancer therapy and numerous approaches have been pursued. Targeting of this process may be achieved by preventing the binding of uPA to u......PAR on the cell surface and/or by direct inhibition of the catalytic activity of uPA. Both strategies have been pursued and inhibition of these functions has shown effect in xenogenic cancer models. Pericellular proteolysis has also been inhibited in vivo in mouse models of wound healing and hepatic fibrinolysis...

  13. Stabilizing a Flexible Interdomain Hinge Region Harboring the SMB Binding Site Drives uPAR into Its Closed Conformation

    DEFF Research Database (Denmark)

    Zhao, Baoyu; Gandhi, Sonu; Yuan, Cai

    2015-01-01

    that the reciprocal stabilization is indeed also possible. By surface plasmon resonance studies we show that these mAbs and vitronectin have overlapping binding sites on uPAR and that they share Arg(91) as hotspot residue in their binding interfaces. The crystal structure solved for one of these uPAR•mAb complexes...... of the conformational status of uPAR and its occupancy with uPA. This is to the best of our knowledge the first study, showing that the dynamic assembly of the three LU domains in uPAR(wt) can be driven towards the closed form by an external ligand, which is not engaging the hydrophobic uPA-binding cavity...

  14. uPA binding sites on domain 2+3 on uPAR and antibodies reactive therewith

    DEFF Research Database (Denmark)

    1996-01-01

    Sammlung von Mikroorganismen und Zellkulturen (DSM) with accession numbers DSM ACC2178 and DSM ACC2179 under the terms and conditions of the Budapest Treaty. The invention furthermore relates to a method for detecting or quantifying u-PAR or a glycosylation variant of u-PAR in a sample by use...... of an antibody according to the invention. Furthermore, the invention relates to a method for the manufacture of a therapeutic agent for preventing or counteracting localized proteolytic activity using of an antibody of the invention as well as to use of an antibody according to the invention for the preparation...

  15. Rational targeting of the urokinase receptor (uPAR): development of antagonists and non-invasive imaging probes

    DEFF Research Database (Denmark)

    Kriegbaum, Mette Camilla; Persson, Morten; Haldager, L

    2011-01-01

    In the last two decades, the urokinase-type plasminogen activator receptor (uPAR) has been implicated in a number of human pathologies such as cancer, bacterial infections, and paroxysmal nocturnal hemoglobinuria. The primary function of this glycolipid-anchored receptor is to focalize uPA-mediat......In the last two decades, the urokinase-type plasminogen activator receptor (uPAR) has been implicated in a number of human pathologies such as cancer, bacterial infections, and paroxysmal nocturnal hemoglobinuria. The primary function of this glycolipid-anchored receptor is to focalize u...

  16. Urokinase-type plasminogen activator receptor (uPAR) expression is associated with T-stage and survival in urothelial carcinoma of the bladder

    DEFF Research Database (Denmark)

    Dohn, Line Hammer; Illemann, Martin; Høyer-Hansen, Gunilla

    2015-01-01

    , and myofibroblasts at the invasive front and tumor core, respectively. Statistical analyses were performed to evaluate the association of uPAR localization and score with clinicopathologic covariates and survival. RESULTS: uPAR positivity was seen in 122/137 (89%) and 118/149 (74%) of the neoplasias at the invasive...... front and tumor core, respectively. uPAR was primarily expressed by myofibroblasts and macrophages in the surrounding stroma as well as some cancer cells. A significant association between uPAR positivity and T-stage as well as grade was found for all 3 cell types in tumor core (P ≤ 0.04 for all...

  17. uPAR-targeted optical near-infrared (NIR) fluorescence imaging and PET for image-guided surgery in head and neck cancer

    DEFF Research Database (Denmark)

    Christensen, Anders; Juhl, Karina; Persson, Morten

    2017-01-01

    . Histological analysis showed co-localization of the fluorescent signal, uPAR expression and tumor deposits. In addition, the feasibility of uPAR-guided robotic cancer surgery was demonstrated. Also, uPAR-PET imaging showed a clear and localized signal in the tongue tumors. CONCLUSIONS: This study demonstrated...... the feasibility of combining two uPAR-targeted probes in a preclinical head and neck cancer model. The PET modality provided preoperative non-invasive tumor imaging and the optical modality allowed for real-time fluorescence-guided tumor detection and resection. Clinical translation of this platform seems...

  18. Autometallographic (AMG) technique used for enhancement of the Golgi-Cox staining gives good contrast andhigh resolution of dendrites and spines

    DEFF Research Database (Denmark)

    Orlowski, Dariusz

    Despite the existence of many newer staining methods, Golgi staining still remains the primary method forvisualization of the dendrites and spines. The black deposit in the Golgi-Cox impregnated cells is a Mercuricsulphide, therefore autometallographic (AMG) technique which is used for visualizat......Despite the existence of many newer staining methods, Golgi staining still remains the primary method forvisualization of the dendrites and spines. The black deposit in the Golgi-Cox impregnated cells is a Mercuricsulphide, therefore autometallographic (AMG) technique which is used...... for visualization of the metals and metalsulphides/selenides in tissue may be used to enhance the Golgi-Cox staining. We demonstrated accordingly thatuse of AMG enhancement method on the Golgi-Cox staining gives good contrast and high resolution of dendritesand spines. Moreover, this method is cheaper and more...

  19. Enhanced antigen detection in immunohistochemical staining using a 'digitized' chimeric antibody.

    Science.gov (United States)

    Eng, Hui-Yan; Wang, Cheng-I; Xue, Yuezhen; Lee, Chia-Yin; Zulkifli, Sarah Binte; Chiam, Poh-Cheang; Ghadessy, Farid J; Lane, David P

    2016-01-01

    The immunohistochemical (IHC) staining of mouse tissue sections using antibodies of mouse origin can result in high nonspecific background due to the staining of endogenous immunoglobulins (Igs) by enzyme-conjugated secondary antibodies. In order to obviate this issue, we developed a chimeric mouse-human anti-p53 monoclonal antibody (MH242) by grafting the variable regions of a known mouse antibody into a human Ig scaffold. This facilitated use of an anti-human secondary antibody, and resulted in near-zero background when compared with its parental mouse monoclonal antibody (PAb242). Furthermore, the chimeric antibody enabled reproducible detection of mutant p53 (homozygous R172H) expression in mouse tissue, an observation hitherto largely equivocal based on the use of existing antibodies. The approach we describe leads to the generation of tractable antibody reagents, whose integrity can be readily verified through DNA sequencing of expressor plasmids. The wide-spread adoption of such 'digitized' antibodies should reduce experimental disparities that can commonly arise through variations in antibody quality. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  20. A Gram Stain Hands-On Workshop Enhances First Year Medical Students' Technique Competency in Comprehension and Memorization.

    Directory of Open Access Journals (Sweden)

    Matthew S Delfiner

    Full Text Available Laboratory diagnostic tests have an essential role in patient care, and the increasing number of medical and health professions schools focusing on teaching laboratory medicine to pre-clinical students reflects this importance. However, data validating the pedagogical methods that best influence students' comprehension and interpretation of diagnostic tests have not been well described. The Gram stain is a simple yet significant and frequently used diagnostic test in the clinical setting that helps classify bacteria into two major groups, Gram positive and negative, based on their cell wall structure.We used this technique to assess which educational strategies may improve students' learning and competency in medical diagnostic techniques. Hence, in this randomized controlled study, we compared the effectiveness of several educational strategies (e.g. workshop, discussion, or lecture in first year medical students' competency in comprehension and interpretation of the Gram stain procedure. We demonstrated that a hands-on practical workshop significantly enhances students' competency in memorization and overall comprehension of the technique. Interestingly, most students irrespective of their cohort showed difficulty in answering Gram stain-related analytical questions, suggesting that more emphasis should be allocated by the instructors to clearly explain the interpretation of the diagnostic test results to students in medical and health professional schools.This proof of principle study highlights the need of practical experiences on laboratory medical techniques during pre-clinical training to facilitate future medical doctors' and healthcare professionals' basic understanding and competency in diagnostic testing for better patient care.

  1. A Gram Stain Hands-On Workshop Enhances First Year Medical Students' Technique Competency in Comprehension and Memorization

    Science.gov (United States)

    Delfiner, Matthew S.; Martinez, Luis R.; Pavia, Charles S.

    2016-01-01

    Background Laboratory diagnostic tests have an essential role in patient care, and the increasing number of medical and health professions schools focusing on teaching laboratory medicine to pre-clinical students reflects this importance. However, data validating the pedagogical methods that best influence students’ comprehension and interpretation of diagnostic tests have not been well described. The Gram stain is a simple yet significant and frequently used diagnostic test in the clinical setting that helps classify bacteria into two major groups, Gram positive and negative, based on their cell wall structure. Methods and Findings We used this technique to assess which educational strategies may improve students’ learning and competency in medical diagnostic techniques. Hence, in this randomized controlled study, we compared the effectiveness of several educational strategies (e.g. workshop, discussion, or lecture) in first year medical students’ competency in comprehension and interpretation of the Gram stain procedure. We demonstrated that a hands-on practical workshop significantly enhances students’ competency in memorization and overall comprehension of the technique. Interestingly, most students irrespective of their cohort showed difficulty in answering Gram stain-related analytical questions, suggesting that more emphasis should be allocated by the instructors to clearly explain the interpretation of the diagnostic test results to students in medical and health professional schools. Conclusion This proof of principle study highlights the need of practical experiences on laboratory medical techniques during pre-clinical training to facilitate future medical doctors’ and healthcare professionals’ basic understanding and competency in diagnostic testing for better patient care. PMID:27711118

  2. Optimization and enhancement of H&E stained microscopical images by applying bilinear interpolation method on lab color mode.

    Science.gov (United States)

    Kuru, Kaya

    2014-02-06

    Hematoxylin & Eosin (H&E) is a widely employed technique in pathology and histology to distinguish nuclei and cytoplasm in tissues by staining them in different colors. This procedure helps to ease the diagnosis by enhancing contrast through digital microscopes. However, microscopic digital images obtained from this technique usually suffer from uneven lighting, i.e. poor Koehler illumination. Several off-the-shelf methods particularly established to correct this problem along with some popular general commercial tools have been examined to find out a robust solution. First, the characteristics of uneven lighting in pathological images obtained from the H&E technique are revealed, and then how the quality of these images can be improved by employing bilinear interpolation based approach applied on the channels of Lab color mode is explored without losing any essential detail, especially for the color information of nuclei (hematoxylin stained sections). Second, an approach to enhance the nuclei details that are a fundamental part of diagnosis and crucially needed by the pathologists who work with digital images is demonstrated. Merits of the proposed methodology are substantiated on sample microscopic images. The results show that the proposed methodology not only remedies the deficiencies of H&E microscopical images, but also enhances delicate details. Non-uniform illumination problems in H&E microscopical images can be corrected without compromising crucial details that are essential for revealing the features of tissue samples.

  3. Improved PET Imaging of uPAR expression using new Cu-64-labeled cross-bridged peptide ligands

    DEFF Research Database (Denmark)

    Persson, Morten; Hosseini, Masood; Madsen, Jacob

    2013-01-01

    The correlation between uPAR expression, cancer cell invasion and metastases is now well-established and has prompted the development of a number of uPAR PET imaging agents, which could potentially identify cancer patients with invasive and metastatic lesions. In the present study, we synthesized......, the more stable of the new uPAR PET tracers, (64)Cu-CB-TE2A-PA-AE105, exhibits a significantly reduced liver uptake compared to (64)Cu-DOTA-AE105 as well as (64)Cu-CB-TE2A-AE105, (p...... and characterized two new cross-bridged (64)Cu-labeled peptide conjugates for PET imaging of uPAR and performed a head-to-head comparison with the corresponding and more conventionally used DOTA conjugate. Based on in-source laser-induced reduction of chelated Cu(II) to Cu(I), we now demonstrate the following...... ranking with respect to the chemical inertness of their complexed Cu ions: DOTA-AE105 95%) were achieved in all cases by incubation at 95ºC. In vivo, they display identical tumor uptake after 1h, but differ significantly after 22 hrs, where the DOTA-AE105 uptake remains surprisingly high. Importantly...

  4. Plasma levels of intact and cleaved urokinase plasminogen activator receptor (uPAR) in men with clinically localised prostate cancer

    DEFF Research Database (Denmark)

    Kristensen, Gitte; Berg, Kasper Drimer; Lippert, Solvej

    2017-01-01

    analysis and quantified using the areas under the ROC curve (AUC).Results: All soluble uPAR levels were significantly (p=0.03) higher in patients with N1 disease compared with patients with N0/x disease. ROC curves including clinical tumour stage, biopsy Gleason score, prostate-specific antigen and percent...

  5. In vivo antimetastatic effects of uPAR retargeted measles virus in syngeneic and xenograft models of mammary cancer

    Science.gov (United States)

    Jing, Yuqi; Bejarano, Marcela Toro; Zaias, Julia; Merchan, Jaime R.

    2014-01-01

    Purpose The urokinase receptor (uPAR) plays a critical role in breast cancer (BC) progression and metastases, and is a validated target for novel therapies. The current study investigates the effects of MV-uPA, an oncolytic measles virus fully retargeted against uPAR in syngeneic and xenograft BC metastases models. Methods In vitro replication and cytotoxicity of MVs retargeted against human (MV-h-uPA) or mouse (MV-m-uPA) uPAR were assessed in human and murine cancer and non-cancer mammary epithelial cells. The in vivo effects of species-specific uPAR retargeted MVs were assessed in syngeneic and xenograft models of experimental metastases, established by intravenous administration of luciferase expressing 4T1 or MDA-MD-231 cells. Metastases progression was assessed by in vivo bioluminescence imaging. Tumor targeting was evaluated by qRT-PCR of MV-N, rescue of viable viral particles and immunostaining of MV particles in lungs from tumor bearing mice. Results In vitro, MV-h-uPA and MV-m-uPA selectively infected, replicated and induced cytotoxicity in cancer compared to non-cancer cells in a species-specific manner. In vivo, MV-m-uPA delayed 4T1 lung metastases progression and prolonged survival. These effects were associated with identification of viable viral particles, viral RNA and detection of MV-N by immunostaining from lung tissues in treated mice. In the human MDA-MB-231 metastases model, intravenous administration of MV-h-uPA markedly inhibited metastases progression and significantly improved survival, compared to controls. No significant treatment related toxicity was observed in treated mice. Conclusions The above preclinical findings strongly suggest that uPAR retargeted measles virotherapy is a novel and feasible systemic therapy strategy against metastatic breast cancer. PMID:25519042

  6. Mapping the topographic epitope landscape on the urokinase plasminogen activator receptor (uPAR) by surface plasmon resonance and X-ray crystallography

    DEFF Research Database (Denmark)

    Zhao, Baoyu; Gandhi, Sonu; Yuan, Cai

    2015-01-01

    ]) and to the general mapping of the topographic epitope landscape on uPAR. The methods required to achieve these data include: (1) recombinant expression and purification of a uPAR-hybrid protein trapped in the desired conformation [patent; WO 2013/020898 A12013]; (2) developing monoclonal antibodies with unique...

  7. Selective abrogation of the uPA-uPAR interaction in vivo reveals a novel role in suppression of fibrin-associated inflammation

    DEFF Research Database (Denmark)

    Connolly, Brian M; Choi, Eun Young; Gårdsvoll, Henrik

    2010-01-01

    of the domain. Analysis of Plau(GFDhu/GFDhu) mice revealed an unanticipated role of the uPA-uPAR interaction in suppressing inflammation secondary to fibrin deposition. In contrast, leukocyte recruitment and tissue regeneration were unaffected by the loss of uPA binding to uPAR. This study identifies...

  8. Development and preclinical evaluation of a new viewing filter system to control reflection and enhance dye staining during vitrectomy.

    Science.gov (United States)

    Enaida, Hiroshi; Hachisuka, Yoshiyuki; Yoshinaga, Yukiyasu; Ikeda, Yasuhiro; Hisatomi, Toshio; Yoshida, Shigeo; Oshima, Yusuke; Kadonosono, Kazuaki; Ishibashi, Tatsuro

    2013-02-01

    We developed a new artificial image enhancement system and evaluated its usefulness in controlling intraoperative reflection and enhancing of Brilliant Blue G (BBG) staining. The system was composed of three kinds of filters (a polarizing filter, a blue-enhancing filter, and a sharp-cut filter Y) and attached to the inferior surface of the operating microscope. Twenty-seven post-mortem extracted porcine eyes were used for a series of examinations. We performed surgery using the 23G-vitrectomy system with a halogen light and xenon lights and compared the reduction of intraoperative reflection under air condition and visibility and BBG contrast with and without this system. The evaluation of images was calculated in CIE 1976 (L*, a*, b*) color space (CIELAB) carried out by ImageJ software. The transmission of each filter and absorbance of BBG was measured by a spectrophotometer. We measured spectral irradiance at each wavelength about each filter from each light source with a spectroradiometer. Under both light sources, intraoperative reflection was controlled using a polarizing (PL) filter or combination of filters under air condition. Evaluation of the value of L* within the cutter surface was changed by 37.8 % under the halogen light, and 61.6 % (averaged) under the xenon light with inserted filters versus no filter. The BBG intensity difference was obtained with sharp-cut Y filter under both light source and PL with blue enhancing filter under the halogen light using each L*, a*, b* parameter with statistically significant (p system can reduce intraoperative reflections under the air condition and obtain an excellent BBG staining intensity induced by various light sources.

  9. Selective abrogation of the uPA-uPAR interaction in vivo reveals a novel role in suppression of fibrin-associated inflammation

    DEFF Research Database (Denmark)

    Connolly, Brian M; Choi, Eun Young; Gårdsvoll, Henrik

    2010-01-01

    the interaction between endogenous uPA and uPAR is selectively abrogated, whereas other functions of both the protease and its receptor are retained. Specifically, we introduced 4 amino acid substitutions into the growth factor domain (GFD) of uPA that abrogate uPAR binding while preserving the overall structure...... of the domain. Analysis of Plau(GFDhu/GFDhu) mice revealed an unanticipated role of the uPA-uPAR interaction in suppressing inflammation secondary to fibrin deposition. In contrast, leukocyte recruitment and tissue regeneration were unaffected by the loss of uPA binding to uPAR. This study identifies...... a principal in vivo role of the uPA-uPAR interaction in cell-associated fibrinolysis critical for suppression of fibrin accumulation and fibrin-associated inflammation and provides a valuable model for further exploration of this multifunctional receptor....

  10. Factor XII and uPAR upregulate neutrophil functions to influence wound healing.

    Science.gov (United States)

    Stavrou, Evi X; Fang, Chao; Bane, Kara L; Long, Andy T; Naudin, Clément; Kucukal, Erdem; Gandhi, Agharnan; Brett-Morris, Adina; Mumaw, Michele M; Izadmehr, Sudeh; Merkulova, Alona; Reynolds, Cindy C; Alhalabi, Omar; Nayak, Lalitha; Yu, Wen-Mei; Qu, Cheng-Kui; Meyerson, Howard J; Dubyak, George R; Gurkan, Umut A; Nieman, Marvin T; Sen Gupta, Anirban; Renné, Thomas; Schmaier, Alvin H

    2018-01-29

    Coagulation factor XII (FXII) deficiency is associated with decreased neutrophil migration, but the mechanisms remain uncharacterized. Here, we examine how FXII contributes to the inflammatory response. In 2 models of sterile inflammation, FXII-deficient mice (F12-/-) had fewer neutrophils recruited than WT mice. We discovered that neutrophils produced a pool of FXII that is functionally distinct from hepatic-derived FXII and contributes to neutrophil trafficking at sites of inflammation. FXII signals in neutrophils through urokinase plasminogen activator receptor-mediated (uPAR-mediated) Akt2 phosphorylation at S474 (pAktS474). Downstream of pAkt2S474, FXII stimulation of neutrophils upregulated surface expression of αMβ2 integrin, increased intracellular calcium, and promoted extracellular DNA release. The sum of these activities contributed to neutrophil cell adhesion, migration, and release of neutrophil extracellular traps in a process called NETosis. Decreased neutrophil signaling in F12-/- mice resulted in less inflammation and faster wound healing. Targeting hepatic F12 with siRNA did not affect neutrophil migration, whereas WT BM transplanted into F12-/- hosts was sufficient to correct the neutrophil migration defect in F12-/- mice and restore wound inflammation. Importantly, these activities were a zymogen FXII function and independent of FXIIa and contact activation, highlighting that FXII has a sophisticated role in vivo that has not been previously appreciated.

  11. Localization of uPAR and MMP-9 in lipid rafts is critical for migration, invasion and angiogenesis in human breast cancer cells

    Directory of Open Access Journals (Sweden)

    Estes Norman

    2010-11-01

    Full Text Available Abstract Background uPAR and MMP-9, which play critical roles in tumor cell invasion, migration and angiogenesis, have been shown to be associated with lipid rafts. Methods To investigate whether cholesterol could regulate uPAR and MMP-9 in breast carcinoma, we used MβCD (methyl beta cyclodextrin, which extracts cholesterol from lipid rafts to disrupt lipid rafts and studied its effect on breast cancer cell migration, invasion, angiogenesis and signaling. Results Morphological evidence showed the association of uPAR with lipid rafts in breast carcinoma cells. MβCD treatment significantly reduced the colocalization of uPAR and MMP-9 with lipid raft markers and also significantly reduced uPAR and MMP-9 at both the protein and mRNA levels. Spheroid migration and invasion assays showed inhibition of breast carcinoma cell migration and invasion after MβCD treatment. In vitro angiogenesis studies showed a significant decrease in the angiogenic potential of cells pretreated with MβCD. MβCD treatment significantly reduced the levels of MMP-9 and uPAR in raft fractions of MDA-MB-231 and ZR 751 cells. Phosphorylated forms of Src, FAK, Cav, Akt and ERK were significantly inhibited upon MβCD treatment. Increased levels of soluble uPAR were observed upon MβCD treatment. Cholesterol supplementation restored uPAR expression to basal levels in breast carcinoma cell lines. Increased colocalization of uPAR with the lysosomal marker LAMP1 was observed in MβCD-treated cells when compared with untreated cells. Conclusion Taken together, our results suggest that cholesterol levels in lipid rafts are critical for the migration, invasion, and angiogenesis of breast carcinoma cells and could be a critical regulatory factor in these cancer cell processes mediated by uPAR and MMP-9.

  12. Mapping the topographic epitope landscape on the urokinase plasminogen activator receptor (uPAR by surface plasmon resonance and X-ray crystallography

    Directory of Open Access Journals (Sweden)

    Baoyu Zhao

    2015-12-01

    Full Text Available The urokinase-type plasminogen activator receptor (uPAR or CD87 is a glycolipid-anchored membrane protein often expressed in the microenvironment of invasive solid cancers and high levels are generally associated with poor patient prognosis (Kriegbaum et al., 2011 [1]. uPAR is organized as a dynamic modular protein structure composed of three homologous Ly6/uPAR domains (LU.This internally flexible protein structure of uPAR enables an allosteric regulation of the interactions with its two principal ligands: the serine protease urokinase-type plasminogen activator (uPA and the provisional matrix protein vitronectin (Vn (Mertens et al., 2012; Gårdsvoll et al., 2011; Madsen et al., 2007 [2–4]. The data presented here relates to the non-covalent trapping of one of these biologically relevant uPAR-conformations by a novel class of monoclonal antibodies (Zhao et al., 2015 [5] and to the general mapping of the topographic epitope landscape on uPAR. The methods required to achieve these data include: (1 recombinant expression and purification of a uPAR-hybrid protein trapped in the desired conformation [patent; WO 2013/020898 A12013]; (2 developing monoclonal antibodies with unique specificities using this protein as antigen; (3 mapping the functional epitope on uPAR for these mAbs by surface plasmon resonance with a complete library of purified single-site uPAR mutants (Zhao et al., 2015; Gårdsvoll et al., 2006 [5,6]; and finally (4 solving the three-dimensional structures for one of these mAbs by X-ray crystallography alone and in complex with uPAR [deposited in the PDB database as 4QTH and 4QTI, respectively].

  13. Advances in Automated Plankton Imaging: Enhanced Throughput, Automated Staining, and Extended Deployment Modes for Imaging FlowCytobot

    Science.gov (United States)

    Sosik, H. M.; Olson, R. J.; Brownlee, E.; Brosnahan, M.; Crockford, E. T.; Peacock, E.; Shalapyonok, A.

    2016-12-01

    Imaging FlowCytobot (IFCB) was developed to fill a need for automated identification and monitoring of nano- and microplankton, especially phytoplankton in the size range 10 200 micrometer, which are important in coastal blooms (including harmful algal blooms). IFCB uses a combination of flow cytometric and video technology to capture high resolution (1 micrometer) images of suspended particles. This proven, now commercially available, submersible instrument technology has been deployed in fixed time series locations for extended periods (months to years) and in shipboard laboratories where underway water is automatically analyzed during surveys. Building from these successes, we have now constructed and evaluated three new prototype IFCB designs that extend measurement and deployment capabilities. To improve cell counting statistics without degrading image quality, a high throughput version (IFCB-HT) incorporates in-flow acoustic focusing to non-disruptively pre-concentrate cells before the measurement area of the flow cell. To extend imaging to all heterotrophic cells (even those that do not exhibit chlorophyll fluorescence), Staining IFCB (IFCB-S) incorporates automated addition of a live-cell fluorescent stain (fluorescein diacetate) to samples before analysis. A horizontally-oriented IFCB-AV design addresses the need for spatial surveying from surface autonomous vehicles, including design features that reliably eliminate air bubbles and mitigate wave motion impacts. Laboratory evaluation and test deployments in waters near Woods Hole show the efficacy of each of these enhanced IFCB designs.

  14. Myofibre segmentation in H&E stained adult skeletal muscle images using coherence-enhancing diffusion filtering.

    Science.gov (United States)

    Strange, Harry; Scott, Ian; Zwiggelaar, Reyer

    2014-10-29

    The correct segmentation of myofibres in histological muscle biopsy images is a critical step in the automatic analysis process. Errors occurring as a result of incorrect segmentations have a compounding effect on latter morphometric analysis and as such it is vital that the fibres are correctly segmented. This paper presents a new automatic approach to myofibre segmentation in H&E stained adult skeletal muscle images that is based on Coherence-Enhancing Diffusion filtering. The procedure can be broadly divided into four steps: 1) pre-processing of the images to extract only the eosinophilic structures, 2) performing of Coherence-Enhancing Diffusion filtering to enhance the myofibre boundaries whilst smoothing the interior regions, 3) morphological filtering to connect unconnected boundary regions and remove noise, and 4) marker controlled watershed transform to split touching fibres. The method has been tested on a set of adult cases with a total of 2,832 fibres. Evaluation was done in terms of segmentation accuracy and other clinical metrics. The results show that the proposed approach achieves a segmentation accuracy of 89% which is a significant improvement over existing methods.

  15. Perilesional Inflammation in Neurocysticercosis - Relationship Between Contrast-Enhanced Magnetic Resonance Imaging, Evans Blue Staining and Histopathology in the Pig Model.

    Science.gov (United States)

    Cangalaya, Carla; Bustos, Javier A; Calcina, Juan; Vargas-Calla, Ana; Suarez, Diego; Gonzalez, Armando E; Chacaltana, Juan; Guerra-Giraldez, Cristina; Mahanty, Siddhartha; Nash, Theodore E; García, Hector H

    2016-07-01

    Disease manifestations in neurocysticercosis (NCC) are frequently due to inflammation of degenerating Taenia solium brain cysts. Exacerbated inflammation post anthelmintic treatment is associated with leakage of the blood brain barrier (BBB) using Evans blue (EB) staining. How well EB extravasation into the brain correlates with magnetic resonance imaging (MRI) using gadolinium (Gd) enhancement as a contrast agent and pericystic inflammation was analyzed in pigs harboring brain cysts of Taenia solium. Three groups of 4 naturally infected pigs were assessed. The first and second groups were treated with both praziquantel plus albendazole and sacrificed two and five days post treatment, respectively. A third untreated group remained untreated. Pigs were injected with EB two hours prior to evaluation by Gd-enhanced T1-MRI, and euthanized. The EB staining for each cyst capsule was scored (EB grades were 0: 0%; 1: up to 50%; 2: over 50% but less than 100%; 3: 100%). Similarly, the Gd enhancement around each cyst was qualitatively and quantitatively scored from the MRI. The extent of pericystic inflammation on histology was scored in increasing severity as IS1, IS2, IS3 and IS4. Grade 3 EB staining and enhancement was only seen in treated capsules. Also, treated groups had higher Gd intensity than the untreated group. Grades of enhancement correlated significantly with Gd enhancement intensity. EB staining was correlated with Gd enhancement intensity and with IS4 in the treated groups. These correlations were stronger in internally located cysts compared to superficial cysts in treated groups. EB staining and Gd enhancement strongly correlate. The intensity of enhancement determined by MRI is a good indication of the degree of inflammation. Similarly, EB staining highly correlates with the degree of inflammation and may be applied to study inflammation in the pig model of NCC.

  16. Number and brightness image analysis reveals ATF-induced dimerization kinetics of uPAR in the cell membrane.

    Science.gov (United States)

    Hellriegel, Christian; Caiolfa, Valeria R; Corti, Valeria; Sidenius, Nicolai; Zamai, Moreno

    2011-09-01

    We studied the molecular forms of the GPI-anchored urokinase plasminogen activator receptor (uPAR-mEGFP) in the human embryo kidney (HEK293) cell membrane and demonstrated that the binding of the amino-terminal fragment (ATF) of urokinase plasminogen activator is sufficient to induce the dimerization of the receptor. We followed the association kinetics and determined precisely the dimeric stoichiometry of uPAR-mEGFP complexes by applying number and brightness (N&B) image analysis. N&B is a novel fluctuation-based approach for measuring the molecular brightness of fluorophores in an image time sequence in live cells. Because N&B is very sensitive to long-term temporal fluctuations and photobleaching, we have introduced a filtering protocol that corrects for these important sources of error. Critical experimental parameters in N&B analysis are illustrated and analyzed by simulation studies. Control experiments are based on mEGFP-GPI, mEGFP-mEGFP-GPI, and mCherry-GPI, expressed in HEK293. This work provides a first direct demonstration of the dimerization of uPAR in live cells. We also provide the first methodological guide on N&B to discern minor changes in molecular composition such as those due to dimerization events, which are involved in fundamental cell signaling mechanisms.

  17. Wavelength dependence of the time course of fluorescence enhancement and photobleaching during irradiation of ethidium bromide-stained nuclei

    Directory of Open Access Journals (Sweden)

    L Galassi

    2009-12-01

    Full Text Available The variation of fluorescence during irradiation of ethidium bromide-stained nuclei with the 458 nm argon laser line was measured at different wavelengths throughout the emission spectrum. When glycerol was used as a mountant, photoenhancement of fluorescence was observed at all wavelengths, but was greater at the shorter wavelengths. Fluorescence increased by almost one order of magnitude at 500 nm after 40 s of irradiation, compared with only about 10% at wavelengths longer than 600 nm after 2-3 s. In nuclei mounted in phosphate buffer, an initial photoenhancement of fluorescence was detected only at the shorter wavelengths, while continuous photobleaching was observed in the rest of the emission spectrum. When the spectra are normalized to maximum, so as to eliminate the effect of the concurrent photobleaching, it appears that the difference between the time course of fluorescence variation in buffer and glycerol depends largely on the lower photobleaching rate in glycerol. The photoenhancement of fluorescence at shorter wavelengths was found to consist of a band peaking at 485-491 nm in glycerol and at 495-496 nm in buffer. Attenuation of the inner-filter effect contributes minimally to the enhancement of fluores- cence at shorter wavelengths. Since the dimer is known to be non fluorescent, the light-induced disaggregation of dimers to monomers cannot be an explanation for the large increase of fluorescence at the shorter wavelengths. The same laser beam that was used to excite the fluorescence of stained nuclei was also used for monitoring the concomitant variation of transmitted light, from which the variation of absorptance during irradiation was computed. While the expected decrease of absorptance was observed in glycerol, reflecting the photodestruction of the fluorophore, in buffer solution an unexpected initial increase was found, which may reflect the accumulation of an absorbing photoproduct.

  18. Enhancing Students` Speaking Skill through Task-Based Language Teaching (TBLT at English Tadris Department of STAIN Kerinci

    Directory of Open Access Journals (Sweden)

    Heri Mudra

    2016-02-01

    Full Text Available This paper reports a classroom action research which conducted in an EFL classroom. The problem of this study is that teaching and learning process tends to be monotonous due to the single method used by English teachers. The learners` speaking course is familiarized with English structures. It requires a communicative and constructive method such as TBLT. The purpose of this study is to describe the effectiveness of TBLT in enhancing students` speaking skill. 30 EFL learners at the seventh semester at STAIN Kerinci took a part in this study. The instruments used to collect the data were speaking test, observation, and field-note. The results of the study showed that there were 2 cycles needed to implement the method. The process of teaching and learning in the first cycle indicates that TBLT improved learners` speaking skill, though some problems were needed to be solved. Unlike the cycle I, the process in the cycle II was improved in term of learners` speaking score and their motivation to attend the course if compared with those in cycle I. So, it is concluded that TBLT is an appropriate method to improve learners` speaking skill.

  19. Group A streptococcal surface GAPDH, SDH, recognizes uPAR/CD87 as its receptor on the human pharyngeal cell and mediates bacterial adherence to host cells.

    Science.gov (United States)

    Jin, Hong; Song, Youngmia P; Boel, Gregory; Kochar, Jaspreet; Pancholi, Vijay

    2005-07-01

    Streptococcal surface dehydrogenase (SDH) is a multifunctional, anchorless protein present on the surface of group A Streptococcus (GAS). It plays a regulatory role in GAS-mediated intracellular signaling events in human pharyngeal cells. Using ligand-binding assays, we have identified an approximately 55 kDa protein as an SDH-specific receptor protein on the surface of Detroit human pharyngeal cells. LC-MS/MS analyses identified this SDH-binding pharyngeal cell-surface-exposed membrane-bound protein as uPAR (urokinase plasminogen activator receptor)/CD87. Ligand-binding assays also revealed that only the N-terminal domain (D1) of uPAR bound to SDH. uPAR-D1 more specifically bound to the C-terminal alpha-helix and two immediate flanking regions of the S-loop of the SDH molecule. Site-directed mutagenesis in GAS resulting in SDH with altered C-terminal ends, and the removal of uPAR from pharyngeal cells by phosphatidylinositol-phopsholipase C treatment decreased GAS ability to adhere to pharyngeal cells. When compared to uninfected Detroit pharyngeal cells, GAS-infected pharyngeal cells showed a transient but a significant increase in the expression of uPAR-specific mRNA, and a prolonged recycling process of uPAR on the cell surface. Together, these results indicate that the specific streptococcal surface protein-pharyngeal cell receptor interaction mediated by SDH and uPAR is modulated during GAS infection of human pharyngeal cells. This interaction significantly contributes to bacterial adherence and thus may play a significant role in GAS pathogenesis by regulating intracellular signaling events in pharyngeal cells.

  20. Inflammation and uPAR-Expression in Colorectal Liver Metastases in Relation to Growth Pattern and Neo-adjuvant Therapy

    DEFF Research Database (Denmark)

    Eefsen, R L; Engelholm, L; Alpizar-Alpizar, W

    2015-01-01

    Proteolytic activity and inflammation in the tumour microenvironment affects cancer progression. In colorectal cancer (CRC) liver metastases it has been observed that three different immune profiles are present, as well as proteolytic activity, determined by the expression of urokinase-type plasm......Proteolytic activity and inflammation in the tumour microenvironment affects cancer progression. In colorectal cancer (CRC) liver metastases it has been observed that three different immune profiles are present, as well as proteolytic activity, determined by the expression of urokinase......-type plasminogen activator (uPAR).The main objectives of this study were to investigate uPAR expression and the density of macrophages (CD68) and T cells (CD3) as markers of inflammation in resected CRC liver metastases, where patients were neo-adjuvantly treated with chemotherapy with or without the angiogenesis...... inhibitor bevacizumab. Chemonaive patients served as a control group. The markers were correlated to growth patterns (GP) of liver metastases, i.e. desmoplastic, pushing and replacement GP. It was hypothesised that differences in proteolysis and inflammation could reflect tumour specific growth and therapy...

  1. Estradiol attenuates EGF-induced rapid uPAR mobilization and cell migration via the G-protein-coupled receptor 30 in ovarian cancer cells

    DEFF Research Database (Denmark)

    Henic, Emir; Noskova, Vera; Høyer-Hansen, Gunilla

    2009-01-01

    and cell migration in ovarian cancer cells and further to identify the ER involved.We used 7 ovarian cancer cell lines, cell migration assay, cellular binding of (125)I-uPA, cellular degradation of (125)I-uPA/PAI-1 complex, enzyme-linked immunosorbent assay for uPAR, solid-phase enzyme immunoassay...

  2. Port-Wine Stains

    Science.gov (United States)

    ... for the Flu Vaccine? Eating Disorders Arrhythmias Port-Wine Stains KidsHealth > For Parents > Port-Wine Stains Print ... Manchas de vino de oporto What Are Port-Wine Stains? A port-wine stain is a type ...

  3. The uPA/uPAR system regulates the bioavailability of PDGF-DD: implications for tumour growth.

    Science.gov (United States)

    Ehnman, M; Li, H; Fredriksson, L; Pietras, K; Eriksson, U

    2009-01-29

    Members of the platelet-derived growth factor (PDGF) family are mitogens for cells of mesenchymal origin and have important functions during embryonic development, blood vessel maturation, fibrotic diseases and cancer. In contrast to the two classical PDGFs, the novel and less well-characterized members, PDGF-CC and PDGF-DD, are latent factors that need to be processed extracellularly by activating proteases, before they can mediate PDGF receptor activation. Here, we elucidate the structural requirements for urokinase plasminogen activator (uPA)-mediated activation of PDGF-DD, as well as the intricate interplay with uPA receptor (uPAR) signalling. Furthermore, we show that activated PDGF-DD, in comparison to latent, more potently transforms NIH/3T3 cells in vitro. Conversely, xenograft studies in nude mice demonstrate that cells expressing latent PDGF-DD are more tumorigenic than those expressing activated PDGF-DD. These findings imply that a fine-tuned proteolytic activation, in the local milieu, controls PDGF-DD bioavailability. Moreover, we suggest that proteolytic activation of PDGF-DD reveals a retention motif mediating interactions with pericellular components. Our proposed mechanism, where uPA not only generates active PDGF-DD, but also regulates its spatial distribution, provides novel insights into the biological function of PDGF-DD.

  4. Dosimetry of 64Cu-DOTA-AE105, a PET tracer for uPAR imaging

    DEFF Research Database (Denmark)

    Persson, Morten; El Ali, Henrik H.; Binderup, Tina

    2014-01-01

    were then exported to OLINDA software for computation of the human absorbed doses. The residence times as well as effective dose equivalent for male and female could be obtained for each organ. To validate this approach, of human projection using mouse data, five mice received iv tail injection......64Cu-DOTA-AE105 is a novel positron emission tomography (PET) tracer specific to the human urokinase-type plasminogen activator receptor (uPAR). In preparation of using this tracer in humans, as a new promising method to distinguish between indolent and aggressive cancers, we have performed PET...... the heart wall to receive the highest dose (0.0918mSv/MBq) followed by the liver (0.0815mSv/MBq), All other organs/tissue were estimated to receive doses in the range of 0.02–0.04mSv/MBq. The mean effective whole-body dose of 64Cu-DOTA-AE105 was estimated to be 0.0317mSv/MBq. Relatively good correlation...

  5. Structure-driven design of radionuclide tracers for non-invasive imaging of uPAR and targeted radiotherapy. The tale of a synthetic peptide antagonist

    DEFF Research Database (Denmark)

    Ploug, Michael

    2013-01-01

    as for monitoring the effects of such treatments by non-invasive imaging using e.g. positron emission tomography. This mini-review will focus on recent advancements in translational research devoted to non-invasive targeting of uPAR, with a view to molecular imaging of its expression in live individuals as well...... as specific eradication of these cells by targeted radiotherapy....

  6. Optimization of construct design and fermentation strategy for the production of bioactive ATF-SAP, a saporin based anti-tumoral uPAR-targeted chimera.

    Science.gov (United States)

    Errico Provenzano, Alfredo; Posteri, Riccardo; Giansanti, Francesco; Angelucci, Francesco; Flavell, Sopsamorn U; Flavell, David J; Fabbrini, Maria Serena; Porro, Danilo; Ippoliti, Rodolfo; Ceriotti, Aldo; Branduardi, Paola; Vago, Riccardo

    2016-11-14

    The big challenge in any anti-tumor therapeutic approach is represented by the development of drugs selectively acting on the target with limited side effects, that exploit the unique characteristics of malignant cells. The urokinase (urokinase-type plasminogen activator, uPA) and its receptor uPAR have been identified as preferential target candidates since they play a key role in the evolution of neoplasms and are associated with neoplasm aggressiveness and poor clinical outcome in several different tumor types. To selectively target uPAR over-expressing cancer cells, we prepared a set of chimeric proteins (ATF-SAP) formed by the human amino terminal fragments (ATF) of uPA and the plant ribosome inactivating protein saporin (SAP). Codon-usage optimization was used to increase the expression levels of the chimera in the methylotrophic yeast Pichia pastoris. We then moved the bioprocess to bioreactors and demonstrated that the fed-batch production of the recombinant protein can be successfully achieved, obtaining homogeneous discrete batches of the desired constructs. We also determined the cytotoxic activity of the obtained batch of ATF-SAP which was specifically cytotoxic for U937 leukemia cells, while another construct containing a catalytically inactive mutant form of SAP showed no activity. Our results demonstrate that the uPAR-targeted, saporin-based recombinant fusion ATF-SAP can be produced in a fed-batch fermentation with full retention of the molecules selective cytotoxicity and hence therapeutic potential.

  7. Pericardial fluid Gram stain

    Science.gov (United States)

    ... a smear. A series of special stains are applied to the sample. This is called a Gram stain . A laboratory specialist looks at the stained slide under the microscope, checking for bacteria. The color, size, and shape of the cells ...

  8. Valor diagnóstico de las proteínas uPAR en sangre para el cáncer gástrico en Guatemala

    Directory of Open Access Journals (Sweden)

    Irmgardt A. Wellmann

    2017-03-01

    Full Text Available El cáncer gástrico es la neoplasia más frecuente del tubo digestivo, Guatemala posee tasas de incidencia y de mortalidad altas. La infección producida por Helicobacter pylori se ha establecido como una de las fuerzas impulsoras de la carcinogénesis gástrica y se ha determinado que la infección con cepas que expresen el factor de virulencia CagA está asociado con lesiones atróficas y precancerosas. El análisis por reacción en cadena de la polimerasa en tiempo real de biopsias del cuerpo gástrico, ha mostrado un incremento significativo de la expresión del activador del plasminógeno de tipo uroquinasa (uPA, su receptor (uPAR y su inhibidor (PAI-I en pacientes positivos para H. pylori. En esta investigación se planteó determinar el valor diagnóstico de las variantes uPAR en sangre como marcador del riesgo de cáncer gástrico en Guatemala, así como su asociación con la infección de cepas de H. pylori virulentas dentro de la población. Para ello se realizaron pruebas serológicas de H. pylori, CagA y la cuantificación de las proteínas uPAR a los participantes (casos y controles, evidenciándose que la prevalencia de participantes con serología positiva para cepas de H. pylori virulentas asciende a 54.9% y el 18% corresponde a verdaderos negativos para H. pylori cagA negativos. Se demostró que existe diferencia significativa en las medias de las cuantificaciones de uPAR entre casos y controles. La curva ROC mostró que resulta razonable plantear que la cuantificación de uPAR es una prueba diagnóstica con capacidad aceptable de discriminar pacientes con y sin cáncer gástrico.

  9. Comparison of photoacoustically derived hemoglobin and oxygenation measurements with contrast-enhanced ultrasound estimated vascularity and immunohistochemical staining in a breast cancer model.

    Science.gov (United States)

    Eisenbrey, John R; Merton, Daniel A; Marshall, Andrew; Liu, Ji-Bin; Fox, Traci B; Sridharan, Anush; Forsberg, Flemming

    2015-01-01

    In this preliminary study, we compared two noninvasive techniques for imaging intratumoral physiological conditions to immunohistochemical staining in a murine breast cancer model. MDA-MB-231 tumors were implanted in the mammary pad of 11 nude rats. Ultrasound and photoacoustic (PA) scanning were performed using a Vevo 2100 scanner (Visualsonics, Toronto, Canada). Contrast-enhanced ultrasound (CEUS) was used to create maximum intensity projections as a measure of tumor vascularity. PAs were used to determine total hemoglobin signal (HbT), oxygenation levels in detected blood (SO2 Avg), and oxygenation levels over the entire tumor area (SO2 Tot). Tumors were then stained for vascular endothelial growth factor (VEGF), cyclooxygenase-2 (Cox-2), and the platelet endothelial cell adhesion molecule CD31. Correlations between findings were analyzed using Pearson's coefficient. Significant correlation was observed between CEUS-derived vascularity measurements and both PA indicators of blood volume (r = 0.49 for HbT, r = 0.50 for SO2 Tot). Cox-2 showed significant negative correlation with SO2 Avg (r = -0.49, p = 0.020) and SO2 Tot (r = -0.43, p = 0.047), while CD31 showed significant negative correlation with CEUS-derived vascularity (r = -0.47, p = 0.036). However, no significant correlation was observed between VEGF expression and any imaging modality (p > 0.08). Photoacoustically derived HbT and SO2 Tot may be a good indicator of tumor fractional vascularity. While CEUS correlates with CD31 expression, photoacoustically derived SO2 Avg appears to be a better predictor of Cox-2 expression. © The Author(s) 2014.

  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. Joint fluid Gram stain

    Science.gov (United States)

    ... called a smear. Several different colored stains are applied to the sample. The laboratory personnel will look at the stained smear under a microscope to see if bacteria are present. The color, size, and shape of ...

  12. 68Ga-labeling and in vivo evaluation of a uPAR binding DOTA- and NODAGA-conjugated peptide for PET imaging of invasive cancers

    DEFF Research Database (Denmark)

    Persson, Morten; Madsen, Jacob; Østergaard, Søren

    2012-01-01

    INTRODUCTION: The urokinase-type plasminogen activator receptor (uPAR) is a well-established biomarker for tumor aggressiveness and metastatic potential. DOTA-AE105 and DOTA-AE105-NH(2) labeled with (64)Cu have previously been demonstrated to be able to noninvasively monitor uPAR expression using...... positron emission tomography (PET) in human cancer xenograft mice models. Here we introduce (68)Ga-DOTA-AE105-NH(2) and (68)Ga-NODAGA-AE105-NH(2) and evaluate their imaging properties using small-animal PET. METHODS: Synthesis of DOTA-AE105-NH(2) and NODAGA-AE105-NH(2) was based on solid-phase peptide...... peptides and identical to AE105. Labeling of DOTA-AE105-NH(2) and NODAGA-AE105-NH(2) with (68)Ga was done at 95°C and room temperature, respectively. The highest radiochemical yield and purity were obtained using fractionated elution, whereas a negative effect of acetone on labeling efficiency for NODAGA...

  13. Safety, dosimetry and tumor detection ability of 68Ga-NOTA-AE105 - a novel radioligand for uPAR PET imaging

    DEFF Research Database (Denmark)

    Skovgaard, Dorthe; Persson, Morten; Brandt-Larsen, Malene

    2017-01-01

    safety and biodistribution in normal tissues and uptake in tumor lesions. METHODS: Ten patients (6 PC, 2 BC and 2 UBC) received a single intravenous dose of (68)Ga-NOTA-AE105 (154 + 59 MBq; range 48-208 MBq). The biodistribution and radiation dosimetry were assessed by serial PET/CT scans (10 minutes, 1...... and 2 hours p.i.). Safety assessment included measurements of vital signs with regular intervals during the imaging sessions and laboratory blood screening tests performed before and after injection. In a subgroup of patients, the in vivo stability of (68)Ga-NOTA-AE105 was determined in collected blood...... therapeutic and diagnostic strategies. In the study, uPAR Positron Emission Tomography (PET) imaging using a (68)Ga-labelled version of the uPAR targeting peptide (AE105) was investigated in a group of patients with BC, PC and UBC. The aim of this first-in-humans, Phase I, clinical trial was to investigate...

  14. Enhancing pathogen identification in patients with meningitis and a negative Gram stain using the BioFire FilmArray(®) Meningitis/Encephalitis panel.

    Science.gov (United States)

    Wootton, Susan H; Aguilera, Elizabeth; Salazar, Lucrecia; Hemmert, Andrew C; Hasbun, Rodrigo

    2016-04-21

    Meningitis with a negative cerebrospinal (CSF) Gram stain represents a diagnostic and therapeutic challenge. The purpose of our study was to evaluate the performance of the BioFire FilmArray(®) Meningitis/Encephalitis (FA ME) panel in patients presenting with community-acquired meningitis with a negative Gram stain. CSF from 48 patients with community-acquired meningitis with a negative Gram stain admitted to four hospitals in Houston, TX underwent additional testing by the FA ME. FA ME results were compared to results obtained as part of routine evaluation. The panel detected pathogens not previously identified in 11 (22.9 %) of 48, but did not detect pathogens identified by standard technique (West Nile virus, Histoplasma) in 5 (15.2 %) patients. Rapid testing for the most common pathogens causing meningitis will aid in the diagnosis and treatment of patients with meningitis.

  15. 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)

  16. 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...

  17. Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

    Science.gov (United States)

    Shi, S R; Key, M E; Kalra, K L

    1991-06-01

    We describe a new approach for retrieval of antigens from formalin-fixed, paraffin-embedded tissues and their subsequent staining by immunohistochemical techniques. This method of antigen retrieval is based on microwave heating of tissue sections attached to microscope slides to temperatures up to 100 degrees C in the presence of metal solutions. Among 52 monoclonal and polyclonal antibodies tested by this method, 39 antibodies demonstrated a significant increase in immunostaining, nine antibodies showed no change, and four antibodies showed reduced immunostaining. In particular, excellent immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. These results showed that after antigen retrieval: (a) enzyme predigestion of tissues could be omitted; (b) incubation times of primary antibodies could be significantly reduced, or dilutions of primary antibodies could be increased; (c) adequate staining could be achieved in long-term formalin-fixed tissues that failed to stain by conventional methods; and (d) certain antibodies which were typically unreactive with formalin-fixed tissues gave excellent staining.

  18. Tissue Staining (Chromoscopy of the Gastrointestinal Tract

    Directory of Open Access Journals (Sweden)

    M Brian Fennerty

    1999-01-01

    Full Text Available Tissue staining, or chomoscopy, is used as an adjunctive technique during gastrointestinal endoscopy. Chemical agents are applied to the gastrointestinal mucosal surface to identify specific epithelia or to enhance the mucosal surface characteristics of the gastrointestinal epithelium. This aids in the recognition of subtle lesions (ie, polyps or allows directed targeting of biopsies (ie, sprue or Barrett’s esophagus to increase the yield of endoscopic diagnostic accuracy. The four endoscopic tissue-staining techniques in use are vital staining, contrast staining (chromoscopy, reactive staining and tattooing. Some of the agents used for endoscopic tissue staining and the uses of chromoscopy in identifying pathology of the esophagus, stomach, small bowel and colon during endoscopy are discussed.

  19. Fluorescent Europium Chelate Stain

    Science.gov (United States)

    Scaff, W. L.; Dyer, D. L.; Mori, K.

    1969-01-01

    The europium chelate of 4,4,4-trifluoro-1-(2-thienyl)-1,3-butanedione (thenoyl-trifluoroacetone; TTA) is firmly bound to microorganisms. It fluoresces brightly at 613 nm with activation at 340 nm. Cells may be stained with 10−3m chelate in 50% ethyl alcohol, followed by washing with 50% ethyl alcohol. Equal or better stains are produced with 10−3m aqueous europium salt, water wash, and 10−2m aqueous TTA. A noncomplexing buffer should be used to maintain the pH at 6.5 to 6.8. Images PMID:4181107

  20. Shimmering Stained Glass.

    Science.gov (United States)

    Simon, Gail Murray

    1998-01-01

    Presents an art lesson for fifth- and sixth-graders where they create a translucent design of colored cellophane on black paper inspired by the stained-glass windows of the Middle Ages and the artwork of Lewis Comfort Tiffany. Enables the students to become crafts people rather than just observers of the past. (CMK)

  1. 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.

  2. Análise florística e estrutural de sistemas silviagrícolas em Tomé-Açu, Pará

    Directory of Open Access Journals (Sweden)

    Édson Luis Bolfe

    2011-10-01

    Full Text Available O objetivo deste trabalho foi analisar a composição florística e estrutural de sistemas silviagrícolas em Tomé-Açu, Pará. Os dados dendrométricos foram obtidos por inventário em 40 parcelas amostrais, com três unidades cada uma, no total de 120 unidades de 10x10 m. Foi inventariada a média de 1.424,3 indivíduos por hectare, pertencentes a 27 famílias e a 54 espécies. Tendo-se considerado a variabilidade dos estágios vegetativos, os diferentes sistemas silviagrícolas (SAF foram divididos em quatro classes hierárquicas: SAF 1, SAF 2, SAF 3, e SAF 4, para estabelecer um sistema de classificação passível de ser utilizado em outras avaliações de campo e em classificações digitais por meio do sensoriamento remoto. Espécies observadas em outros sistemas da região amazônica também foram relevantes para este estudo, especialmente Theobroma cacao, T. grandiflorum e Euterpe oleracea que, juntas, apresentaram médias de frequência relativa de 51%, densidade relativa de 69,2%, dominância relativa de 50,1% e índice de valor de importância de 56,8%. Os dados médios de diversidade florística, abundância, área basal e valor de importância indicam os sistemas silviagrícolas da região de Tomé-Açu como sistemas de produção com potencial econômico e ambiental, se adotado manejo adequado e racional.

  3. Degradation of internalized αvβ5 integrin is controlled by uPAR bound uPA: effect on β1 integrin activity and α-SMA stress fiber assembly.

    Directory of Open Access Journals (Sweden)

    Lingyan Wang

    Full Text Available Myofibroblasts (Mfs that persist in a healing wound promote extracellular matrix (ECM accumulation and excessive tissue contraction. Increased levels of integrin αvβ5 promote the Mf phenotype and other fibrotic markers. Previously we reported that maintaining uPA (urokinase plasminogen activator bound to its cell-surface receptor, uPAR prevented TGFβ-induced Mf differentiation. We now demonstrate that uPA/uPAR controls integrin β5 protein levels and in turn, the Mf phenotype. When cell-surface uPA was increased, integrin β5 levels were reduced (61%. In contrast, when uPA/uPAR was silenced, integrin β5 total and cell-surface levels were increased (2-4 fold. Integrin β5 accumulation resulted from a significant decrease in β5 ubiquitination leading to a decrease in the degradation rate of internalized β5. uPA-silencing also induced α-SMA stress fiber organization in cells that were seeded on collagen, increased cell area (1.7 fold, and increased integrin β1 binding to the collagen matrix, with reduced activation of β1. Elevated cell-surface integrin β5 was necessary for these changes after uPA-silencing since blocking αvβ5 function reversed these effects. Our data support a novel mechanism by which downregulation of uPA/uPAR results in increased integrin αvβ5 cell-surface protein levels that regulate the activity of β1 integrins, promoting characteristics of the persistent Mf.

  4. Digital staining of pathological tissue specimens using spectral transmittance

    Science.gov (United States)

    Bautista, Pinky A.; Abe, Tokiya; Yamaguchi, Masahiro; Yagi, Yukako; Ohyama, Nagaaki

    2005-04-01

    Staining of tissue specimens is a classical procedure in pathological diagnosis to enhance the contrast between tissue components such that identification and classification of these components can be easily performed. In this paper, a framework for digital staining of pathological specimens using the information derived from the L-band spectral transmittance of various pathological tissue components is introduced, particularly the transformation of a Hematoxylin and Eosin (HE) stained specimen to its Masson-Trichrome (MT) stained counterpart. The digital staining framework involves the classification of tissue components, which are highlighted when the specimen is actually stained with MT stain, e.g. fibrosis, from the HE-stained image; and the linear mapping between specific sets of HE and MT stained transmittance spectra through pseudo-inverse procedure to produce the LxL transformation matrices that will be used to transform the HE stained transmittance to its equivalent MT stained transmittance configuration. To generate the digitally stained image, the decisions of multiple quadratic classifiers are pooled to form the weighting factors for the transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) for the implementation of digital staining in the pathological context.

  5. Effectiveness of unsedated transnasal endoscopy with white-light, flexible spectral imaging color enhancement, and lugol staining for esophageal cancer screening in high-risk patients.

    Science.gov (United States)

    Arantes, Vitor; Albuquerque, Walton; Salles, Jose Maria Porcaro; Freitas Dias, Carlos Alberto; Alberti, Luiz Ronaldo; Kahaleh, Michel; Ferrari, Teresa Cristina Abreu; Coelho, Luiz Gonzaga Vaz

    2013-04-01

    Transnasal endoscopy (TNE) has been proposed to screen for esophageal squamous cell cancer (ESCC) in Asia. This study aimed to assess the feasibility and tolerance of Brazilian patients to undergo unsedated TNE for screening, the prevalence of ESCC in this population, and the effectiveness of white-light endoscopy (WLE) and digital chromoendoscopy [flexible spectral imaging color enhancement (FICE)] to diagnose esophageal neoplasia. This was a diagnostic test study that enrolled patients with head and neck squamous cell cancer (HNSCC) referred to ESCC screening. Patients' tolerance was rated by a numeric pain intensity scale. Interventions included unsedated TNE with WLE and FICE examination of the esophagus, in a tandem manner with blinded operators, followed by lugol chromoscopy. Performance of WLE and FICE for neoplasia detection was compared with the reference standard (lugol chromoscopy plus histology). A total of 106 patients were recruited. TNE was feasible in 99.1%, and 92% of the patients rated the discomfort as absent or minimal. Thirteen ESCC were detected (12.3%), with 10 early cancers (77%). The tests showed an excellent performance and there was no difference between WLE (sensitivity 92.3%, specificity 98.9%, accuracy 98.1%, area under curve 0.995) and FICE (sensitivity 100%, specificity 98.9%, accuracy 99%, area under curve 0.956) for esophageal neoplasia detection. Unsedated TNE is a feasible, well accepted, and efficient diagnostic tool for the screening of ESCC. The elevated rate of esophageal neoplasia strengthens the recommendations to screen patients with HNSCC. The yields of WLE and FICE were similar for ESCC detection.

  6. The Social Function of Staining

    OpenAIRE

    GÜNDÜZ, Alev

    2016-01-01

    The place of staining which is established in today’s cosmetic perception as a creative way forbeauty, charm and attraction, had been split the path of the fed formatting of the social necessity ofthe past. The adventure of staining person who constantly reshapes the area based for needing, initiallypointed to similar meanings in the mind and body of every individual in the society. A personwho finds self-expression using non-verbal superior language by staining; has created a new sourceof id...

  7. Gram stain of skin lesion

    Science.gov (United States)

    Skin lesion gram stain ... skin sore. This procedure is called a skin lesion biopsy . Before the biopsy, your provider will numb ... means bacteria have been found in the skin lesion. Further tests are needed to confirm the results. ...

  8. Retraction: "Down-regulation of uPA and uPAR by 3,3'-diindolylmethane contributes to the inhibition of cell growth and migration of breast cancer cells" by Ahmad et al.

    Science.gov (United States)

    2016-08-01

    The above article, published online on August 19, 2009 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor in Chief, Gary S. Stein, and Wiley Periodicals, Inc. The retraction has been agreed following an investigation from Wayne State University involving the third author and the corresponding author that found Figure 5C to be inappropriately re-used and re-labeled. REFERENCE Ahmad A, Kong D, Wang Z, Sarkar SH, Banerjee S, Sarkar FH. 2009. Down-regulation of uPA and uPAR by 3,3'-diindolylmethane contributes to the inhibition of cell growth and migration of breast cancer cells. J Cell Biochem 108:916-925; doi: 10.1002/jcb.22323. © 2016 Wiley Periodicals, Inc.

  9. Special stains in Mohs surgery.

    Science.gov (United States)

    Miller, Christopher J; Sobanko, Joseph F; Zhu, Xiaodong; Nunnciato, Terri; Urban, Christopher R

    2011-04-01

    The excellent cure rates associated with Mohs micrographic surgery depend on accurate interpretation of complete and high-quality microscopic frozen sections. Reliable interpretation of microscopic slides is only possible if the surgeon can distinguish tumor cells from surrounding normal tissue. By highlighting tumor cells with a chromogen that is visible on light microscopy, immunostaining allows the Mohs surgeon to distinguish tumor from normal cells in these challenging scenarios. This article focuses on practical aspects involving the most commonly used immunostains in dermatologic surgery, including MART-1 for melanocytic neoplasms, cytokeratin stains for keratinocytic neoplasms, and CD34 stains for dermatofibrosarcoma protuberans. Copyright © 2011 Elsevier Inc. All rights reserved.

  10. Modified elastic tissue-Masson trichrome stain.

    Science.gov (United States)

    Garvey, W

    1984-07-01

    A combined elastic tissue-Masson technique is presented which stains elastic fibers of all sizes, nuclei and connective tissue. The modified elastic tissue stain consists of hematoxylin, ferric chloride and Verhoeff's iodine; nuclei and elastic fibers are stained blue-black in six minutes without differentiation. By contrast, cytoplasmic elements are stained red, (Biebrich scarlet-acid fuchsin) and collagen is stained green (light green) or blue (aniline blue). The entire staining procedure takes approximately one hour.

  11. Accelerated staining technique using kitchen microwave oven.

    Science.gov (United States)

    Mukunda, Archana; Narayan, T V; Shreedhar, Balasundhari; Shashidhara, R; Mohanty, Leeky; Shenoy, Sadhana

    2015-01-01

    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. 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. 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. 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.

  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. 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.

  14. 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.

  15. Aspectos econômicos da produção e do risco nos sistemas agroflorestais e nos sistemas tradicionais de produçâo agrícola em tomé-açu, Pará - 2001 a 2003 Economic aspects of production under risk conditions in agroforestry systems and traditional agricultural systems in tomé-açu, Pará - 2001 to 2003

    Directory of Open Access Journals (Sweden)

    Luiz Benedito Varela

    2009-02-01

    Full Text Available Neste artigo, analisaram-se os fatores determinantes da produção dinâmica dos sistemas agroflorestais (SAF e dos sistemas tradicionais de produção agrícola (ST, sob condições de risco, em pequenas e médias unidades produtivas nipo-brasileiras localizadas no Município de Tomé-Açu, Pará, no período de 2001 a 2003. Os resultados indicaram que todos os fatores, exceto a mão-de-obra contratada e as máquinas e equipamentos, afetam diretamente o Valor Bruto da Produção (VBP dos SAF e dos ST; a variável dummy apresentou diferença cumulativa a menor no VBP dos SAF, de um ano para outro. A função de risco estimada apontou que os SAF apresentaram menor risco que os ST, evidenciando-se que a aplicação de insumos era fonte de redução de risco, mas a tecnologia adotada precisa ser adequada, pois se apresenta como fator de aumento de risco nos dois sistemas. Além disso, a dummy indicou que os SAF exibiram menor nível de risco que os ST. Nesse contexto, os resultados deixaram claro, ainda, que os produtores nipo-brasileiros eram avessos ao risco.This article analyzes the determinants of the dynamic production inputs of the agroforestry systems (SAF and traditional agricultural systems (ST under risk conditions, in small and medium farms in Tome-Açu, Pará, by Brazilian-Japanese producers from 2001 to 2003. The dynamic regression model results showed that all the inputs, except labor and machinery, have positive impacts on the current VBP of the two systems analyzed. Furthermore, the dummy variable shows a minor accumulative difference of the SAF's VBP in comparison with ST's VBP, from one period (year to another. The estimated risk function indicates that the use of fertilizer and pesticides reduce the risk level of the two systems, but the current technology needs to be adequate to other production inputs, since it appears as a factor of increased risk to the two systems. In addition, the dummy variable indicates that the SAF

  16. 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.

  17. [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.

  18. 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.

  19. A stain on Italian reforms

    CERN Multimedia

    2001-01-01

    "Italy's principal funding agency has missed an opportunity to enhance the prestige of its institutes. In appointing its first crop of new directors, it has conspicuously avoided some candidates of the highest calibre" (1/2 page).

  20. [Quadri-chromic staining in routine cutaneous histopathology (hematoxylin, eosin, saffron and astra blue)].

    Science.gov (United States)

    Cribier, B; Grosshans, E

    1995-01-01

    The routine histologic stains used in dermatopathology are usually hematoxylin-eosin-safran. In order to stain the mucins, specific dyes are necessary. The purpose of this study was to perform a routine quadrichromic stain which included Astra blue as specific dye of mucins. The applications of this hematoxylin-eosin-safran-Astra blue stain were evaluated after 5 years of systematic use. The quadrichromic stain was realized by Varistain (Shandon). The time of coloration was 15 minutes for Astra blue pH 2.5, 10 minutes for hematoxylin, 45 seconds for eosin and 4 minutes for safran. A total of 56,000 samples were analyzed using the quadrichromic stain. When mucin was present, it was colored in blue and the contrast with the other dyes was enhanced. Astra blue stained specifically the mucin deposits and it did not modify the other colours. The mucin deposits present in primary and secondary cutaneous mucinoses were most often visible after quadrichromic stain. Because of the variability of the mucin composition, the Astra blue did not react with all mucins. This stain was particularly useful in extramammary Paget's disease and in atrophic malignant papulosis. This new stain was easy to perform and allowed a specific coloration of mucin deposits in many cases. The use of four colours therefore improves the results of a routine stain, with a high quality of coloration.

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

    African Journals Online (AJOL)

    BackgroundModified Giemsa staining has been favoured by many researchers because it is easy to perform but, like many other stains, demonstration of the bacteria depends on its morphology. It has been arged in some research circles that some of the organisms in the gastric mucosa may not be true H.pylori.

  2. Stains and Stories: Latent narrative in worn clothing

    OpenAIRE

    Goldsmith, Shelly

    2008-01-01

    Stains and Stories is a series of textile-based installations and limited-edition prints, which form part of an ongoing project to examine perceived latent matter (memories and experiences) in worn clothing. The work seeks to present complex ideas about easily accessible objects (clothes), in order to provoke contemplation about human experience and enhance psychological knowledge. I consulted with Dr. Alison Fendley, Senior Biologist at the Forensic Science Service, in developing approa...

  3. Negative staining and cryo-negative staining of macromolecules and viruses for TEM.

    Science.gov (United States)

    De Carlo, Sacha; Harris, J Robin

    2011-02-01

    In this review we cover the technical background to negative staining of biomolecules and viruses, and then expand upon the different possibilities and limitations. Topics range from conventional air-dry negative staining of samples adsorbed to carbon support films, the variant termed the "negative staining-carbon film" technique and negative staining of samples spread across the holes of holey-carbon support films, to a consideration of dynamic/time-dependent negative staining. For each of these approaches examples of attainable data are given. The cryo-negative staining technique for the specimen preparation of frozen-hydrated/vitrified samples is also presented. A detailed protocol to successfully achieve cryo-negative staining with ammonium molybdate is given, as well as examples of data, which support the claim that cryo-negative staining provides a useful approach for the high-resolution study of macromolecular and viral structure. Copyright © 2009 Elsevier Ltd. All rights reserved.

  4. Janus Green B as a rapid, vital stain for peripheral nerves and chordotonal organs in insects.

    Science.gov (United States)

    Yack, J E

    1993-08-01

    Effective staining of peripheral nerves in live insects is achieved with the vital stain Janus Green B. A working solution of 0.02% Janus Green B in saline is briefly applied to the exposed peripheral nervous system. The stain is then decanted and the dissection flooded with fresh saline, resulting in whole nerves being stained dark blue in contrast to surrounding tissues. This simple and reliable technique is useful in describing the distribution of nerves to their peripheral innervation sites, and in locating small nerve branches for extracellular physiological recordings. The stain is also shown to be useful as a means of enhancing the contrast between scolopale caps and surrounding tissues in chordotonal organs, staining chordotonal organ attachment strands, and the crista acustica (tympanal organ) of crickets and katydids. The advantages of Janus Green B over traditional peripheral nerve strains, in addition to its shortcomings, are discussed.

  5. Bacterial endotoxin enhances colorectal cancer cell adhesion and invasion through TLR-4 and NF-kappaB-dependent activation of the urokinase plasminogen activator system.

    LENUS (Irish Health Repository)

    Killeen, S D

    2009-05-19

    Perioperative exposure to lipopolysaccharide (LPS) is associated with accelerated metastatic colorectal tumour growth. LPS directly affects cells through Toll-like receptor 4 (TLR-4) and the transcription factor NF-kappaB. The urokinase plasminogen activator (u-PA) system is intimately implicated in tumour cell extracellular matrix (ECM) interactions fundamental to tumour progression. Thus we sought to determine if LPS directly induces accelerated tumour cell ECM adhesion and invasion through activation of the u-PA system and to elucidate the cellular pathways involved. Human colorectal tumour cell lines were stimulated with LPS. u-PA concentration, u-PA activity, active u-PA, surface urokinase plasminogen activator receptor (u-PAR) and TLR-4 expression were assessed by ELISA, colorimetric assay, western blot analysis and flow cytometry respectively. In vitro tumour cell vitronectin adhesion and ECM invasion were analysed by vitronectin adhesion assay and ECM invasion chambers. u-PA and u-PAR function was inhibited with anti u-PA antibodies or the selective u-PA inhibitors amiloride or WXC-340, TLR-4 by TLR-4-blocking antibodies and NF-kappaB by the selective NF-kappaB inhibitor SN-50. LPS upregulates u-PA and u-PAR in a dose-dependent manner, enhancing in vitro tumour cell vitronectin adhesion and ECM invasion by >40% (P<0.01). These effects were ameliorated by u-PA and u-PAR inhibition. LPS activates NF-kappaB through TLR-4. TLR-4 and NF-kappaB inhibition ameliorated LPS-enhanced u-PA and u-PAR expression, tumour cell vitronectin adhesion and ECM invasion. LPS promotes tumour cell ECM adhesion and invasion through activation of the u-PA system in a TLR-4- and NF-kappaB-dependent manner.

  6. [Comparison of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection].

    Science.gov (United States)

    Huang, Da-Ke; Zhang, Yu-Xia; Man, Su-Qin; Yu, Fa-Zhi; Shen, Ji-Jia

    2012-08-01

    To compare the effects of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection. A model of hepatic granuloma and fibrosis was established by infecting mice with S. japonicum cercariae, then the hepatic specimens were taken and Van-Gieson staining and Masson trichrome staining were performed. Eventually, the area of granuloma and fibrosis were measured by imaging analysis software. When the time of staining was 3-7 min, there was no significant difference of the fibrosis areas between the two methods (P > 0.05); when the time of staining was more than 10 min, the staining area showed by Masson's staining was significantly larger than that showed by Van-Gieson staining, and the difference was statistically significant (P Masson trichrome staining, therefore Van-Gieson staining is a better method to display collagen.

  7. Intracellular and juxtacellular staining with biocytin.

    Science.gov (United States)

    Wilson, Charles J; Sachdev, R N S

    2004-05-01

    Many physiological studies require microscopic examination of the recorded neuron for identification. This unit describes how intracellular and extracellular recording can be combined with single-neuron staining to enable sequential physiological and morphological studies.

  8. 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

  9. Selection and application of exterior stains for wood

    Science.gov (United States)

    R. Sam. Williams; William C. Feist

    1999-01-01

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

  10. [Exogenous tooth discoloration in children: black stains].

    Science.gov (United States)

    Bandon, D; Chabane-Lemboub, A; Le Gall, M

    2011-12-01

    Black-stains are a coloring frequently met in pediatric dentistry. They can be medically diagnosed as 1-mm borders or unfinished lines formed by a dark exogenous substance which follows the gingival festoon of bet coronary (in cervical third of the crown) temporary teeth and permanent, or they can appear in like points or dark spots. They are caused by bacteria anaerobic chromogenous. The dominant responsible species are actinomyces. Blacks-stains are ferrous depots, formed following a chemical interaction on the surface of the tooth between sulphide of hydrogen (under the effect of the anaerobic bacteria which are producing hydrogen) and the iron contained in the saliva (by a healthy diet) or that released by red blood corpuscles (in case of bloody gums). Black-stains are a shape of characteristic dental plaque by its flora with trend to calcify. It contains an insoluble iron salt with a content raised in calcium and in inorganic phosphor. The coloring Black-stain is a mild pathology and has no incidence on the vitality of the tooth. Certainly these spots are unsightly. The dental surgeon in current practice can deprive them. The pediatrician plays a leading role in the diagnosis and advice to parents and patients affected by these stains. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  11. Campylobacter enteritis: early diagnosis with Gram's stain.

    Science.gov (United States)

    Ho, D D; Ault, M J; Ault, M A; Murata, G H

    1982-10-01

    Campylobacter jejuni has become one of the most important causes of infectious diarrhea in the United States. We examined the utility of Gram's stain of stool for the rapid presumptive diagnosis of Campylobacter enteritis in a large, urban hospital and found that this test has a sensitivity of 43.5% and a specificity of 99.4%. We believe that Gram's stain of stool could be used to direct the early management of up to one half of patients infected with this pathogen.

  12. Solid acid catalysts: Stain and shine

    Science.gov (United States)

    Chen, Peng

    2011-11-01

    Catalyst particles for fluid catalytic cracking are vital for the oil-refinery industry, but their activity is hard to diagnose because of their inter- and intra-particle structural inhomogeneity. With fluorescence confocal microscopy and selective staining, one can now pinpoint the catalytic activity within single catalyst particles from an industrial reactor.

  13. Autofluorescence of routinely hematoxylin and eosin- stained ...

    African Journals Online (AJOL)

    SERVER

    2008-03-04

    Mar 4, 2008 ... ... Hosokawa S, Nagaike K, Tagawa T (2004). A new immunofluoro-staining method using red fluorescence of PerCP on formalin-fixed paraffin-embedded tissues. J. Immunol. Methods, 293: 143-151. Rotomskis R, Streckyte G (2004). Fluorescence diagnostics of tumors. Medicina (Kaunas), 40: 1219-1230.

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

    African Journals Online (AJOL)

    1991-03-16

    Mar 16, 1991 ... T. F. H. G. JACKSON, PH.D. V. GATHIRAM, F,C.P.(S.A.). Department of Medical Microbiology, University of Natal,. Durban. J. VAN DEN ENDE, F.F. PATH. (S.A.). A=pted 30 Mat 1990. have similar sizes,-shapes and staining characteristics to yeasts and other coccidia, diagnostic difficulties can be expected.

  15. 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…

  16. 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

  17. 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

  18. Canker Stain Affects Delaware Sycamores Pest Alert

    Science.gov (United States)

    Alan Iskra; Gary Schwetz; Michael Valenti

    2001-01-01

    An often fatal disease of American sycamore (Platanus occidentalis), known as canker stain, is caused by the fungus, Ceratocystis fimbriata f.sp. platani. This fungus, indigenous to the United States, occurs in urban and forested areas from New Jersey to Georgia and west to Missouri and Louisiana. Other trees affected are the Oriental plane (Platanus orientalis) and...

  19. 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... solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures of synthetic or natural dyes or nondye chemicals in solutions used in staining cells and tissues for diagnostic...

  20. Newer applications of the histological stain prepared from Pterocarpus santalinus.

    Science.gov (United States)

    Sen Gupta, P C; Mukherjee, A K

    1981-03-01

    A histological stain prepared from the heartwood of Pterocarpus santalinus Linn. has been found to be an excellent nuclear stain for various cells of animal and plant origin. As an elastic tissue stain, the results are comparable to standard elastic tissue stains. The striations of voluntary muscle fibers are well shown. The Nissl granules and fibers of cranial nerves in the pons are visualized. When counterstained with light green, it differentially stains muscle and fibrous tissue. The stain can be used as counterstain with certain histochemical procedures with satisfactory results. The preparation and use of this versatile stain are described.

  1. 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

  2. Stain removal and whitening by baking soda dentifrice: A review of literature.

    Science.gov (United States)

    Li, Yiming

    2017-11-01

    Tooth discoloration may be caused by intrinsic or extrinsic stains or a combination of both. There are 2 major approaches to removing the stains, including the chemical mechanism using peroxides for tooth bleaching and the mechanical mechanism using abrasives in prophylactic pastes and dentifrices to remove stains, resulting in a whitening effect. Attempts have also been made to add a low concentration of peroxides to dentifrices to enhance their abrasive cleaning to remove tooth stains. This article provides a review of both in vitro and clinical studies on stain removal and whitening effect of dentifrices containing sodium bicarbonate (baking soda). In recent years, whitening dentifrices have become popular because of little additional effort for use, ease of availability, low cost, and accumulated evidence of clinical efficacy and safety in the literature. Advances in research and technology have led to innovative formulations of dentifrices using baking soda as the sole abrasive or a component of an abrasive system. Baking soda is biologically compatible with acid-buffering capacities, antibacterial at high concentrations, and has a relatively lower abrasivity. The evidence available in the literature indicates that baking soda-based dentifrices are effective and safe for tooth stain removal and consequently whitening. A number of clinical studies have also shown that baking soda-based dentifrices are more effective in stain removal and whitening than some non-baking soda-containing dentifrices with a higher abrasivity. So far, research efforts have mainly focused on stain removal and tooth-whitening efficacy and clinical safety of baking soda dentifrices used with manual toothbrushes, with only a few studies investigating their effects using powered toothbrushes, for which further research is encouraged. As part of a daily oral hygiene practice, baking soda-based dentifrice is a desirable, alternative or additional measure for tooth stain removal and whitening

  3. Isolation, Culture, and Staining of Single Myofibers.

    Science.gov (United States)

    Gallot, Yann Simon; Hindi, Sajedah M; Mann, Aman K; Kumar, Ashok

    2016-10-05

    Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.

  4. Ki-67 Membranous Staining: Biologically Relevant or an Artifact of Multiplexed Immunofluorescent Staining.

    Science.gov (United States)

    Wang, Dan; Pang, Zhengyu; Clarke, Gina M; Nofech-Mozes, Sharon; Liu, Kela; Cheung, Alison M Y; Filkins, Robert J; Yaffe, Martin J

    2016-07-01

    In the process of developing a multiplex of 8 common breast cancer biomarkers (Her2/neu, estrogen receptor, progesterone receptor, Ki-67, aldehyde dehydrogenase-1, NaK-ATPase, cytokeratin 8/18, and myosin smooth muscle) on a single formalin-fixed paraffin-embedded slide using a sequential staining, imaging, and dye bleaching technology developed by General Electric Company, membranous Ki-67 staining was observed and colocalized with Her2/neu staining. Using immunohistochemistry as gold standards, we discovered that membranous Ki-67 was an artifact caused by the binding of cyanine 5-conjugated rabbit polyclonal Ki-67 antibody to a secondary cyanine 3-conjugated donkey anti-rabbit antibody which was previously applied and bound to rabbit Her2/neu antibody in our multiplexing experiment. After blocking with rabbit serum, a successful protocol for 8 biomarker multiplexing without cross-reactivity of antibodies from the same species was developed.

  5. Fat tissue staining and photodynamic/photothermal effects

    Science.gov (United States)

    Tuchin, Valery V.; Altshuler, Gregory B.; Yanina, Irina Yu.; Kochubey, Vyacheslav I.; Simonenko, Georgy V.

    2010-02-01

    Cellulite is considered as a disease of the subcutaneous fat layer that appears mostly in women and consists of changes in fat cell accumulation together with disturbed lymphatic drainage, affecting the external appearance of the skin. The photodynamic and selective photothermal treatments may provide reduction the volume of regional or sitespecific accumulations of subcutaneous adipose tissue on the cellular level. We hypothesize that light irradiation of stained fat tissue at selected temperature leads to fat cell lypolytic activity (the enhancement of lipolysis of cell triglycerides due to expression of lipase activity and cell release of free fat acids (FFAs) due to temporal cell membrane porosity), and cell killing due to apoptosis caused by the induced fat cell stress and/or limited cell necrosis.

  6. [Histochemical staining using silver salts using a microwave oven].

    Science.gov (United States)

    Balaton, A

    1987-01-01

    Some metallic impregnations--Fontana-Masson, Warthin-Starry, Grocott's methenamine silver, Grimelius' and Dieterle's stains have been modified to use a microwave oven. Microwave bombardment markedly reduces the staining times and produces a cleaner background.

  7. 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.

  8. 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...

  9. A Digital Staining Algorithm for Optical Coherence Tomography Images of the Optic Nerve Head

    Science.gov (United States)

    Mari, Jean-Martial; Aung, Tin; Cheng, Ching-Yu; Strouthidis, Nicholas G.; Girard, Michaël J. A.

    2017-01-01

    Purpose To digitally stain spectral-domain optical coherence tomography (OCT) images of the optic nerve head (ONH), and highlight either connective or neural tissues. Methods OCT volumes of the ONH were acquired from one eye of 10 healthy subjects. We processed all volumes with adaptive compensation to remove shadows and enhance deep tissue visibility. For each ONH, we identified the four most dissimilar pixel-intensity histograms, each of which was assumed to represent a tissue group. These four histograms formed a vector basis on which we ‘projected' each OCT volume in order to generate four digitally stained volumes P1 to P4. Digital staining was also verified using a digital phantom, and compared with k-means clustering for three and four clusters. Results Digital staining was able to isolate three regions of interest from the proposed phantom. For the ONH, the digitally stained images P1 highlighted mostly connective tissues, as demonstrated through an excellent contrast increase across the anterior lamina cribrosa boundary (3.6 ± 0.6 times). P2 highlighted the nerve fiber layer and the prelamina, P3 the remaining layers of the retina, and P4 the image background. Further, digital staining was able to separate ONH tissue layers that were not well separated by k-means clustering. Conclusion We have described an algorithm that can digitally stain connective and neural tissues in OCT images of the ONH. Translational Relevance Because connective and neural tissues are considerably altered in glaucoma, digital staining of the ONH tissues may be of interest in the clinical management of this pathology. PMID:28174676

  10. Comparison of verdeluz orange G and modified Gallego stains.

    Science.gov (United States)

    Kunche, A; Kiresur, M A; Ananthaneni, A; Guduru, V S; Puneeth, H K; Bagalad, B

    2017-11-21

    Tumors of the oral cavity include combinations of hard and soft tissues that may be difficult to identify using routine hematoxylin and eosin (H & E) staining. Although combination stains can demonstrate hard and soft tissues, trichrome stains, such as VanGieson and Masson, cannot differentiate dental hard tissues, such as dentin, cementum and osteoid. Modified Gallegos (MGS) and verdeluz orange G-acid fuchsin (VOF) stains can differentiate components of teeth. We used 10 tissue sections of decalcified bone and 10 pathologic tissue sections that contained different calcified tissues including peripheral ossifying fibroma, odontoma, central ossifying fibroma and cemento-ossifying fibroma. Sections were stained with H & E, VOF or MGS. H and E stained both hard tissues pink. VOF stained bone purple-red, cementum red and collagen blue. MGS stained bone green-blue, cementum red and collagen blue. VOF staining intensity and differentiation was better than MGS staining. VOF staining demonstrated hard tissue components distinctly and exhibited good contrast with the surrounding connective tissue. VOF also is a simple, single step, rapid staining procedure.

  11. Histological study on the staining potentials of Aqueous extract of ...

    African Journals Online (AJOL)

    Histological study on the staining potentials of Aqueous extract of Ceratonia Siliqua bark. EE Okpidu, AU Okon, GP Oyadonghan, LA Ogbodo, ECN Onyenekwe. Abstract. This study was designed to determine the staining potentials of aqueous extract of Ceratonia Siliqua bark adapted for the first time as a counter stain in ...

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

    African Journals Online (AJOL)

    SITWALA COMPUTERS

    5 and the cyclospora species. Their life cycle requires an external intermediate host, usually an animal, ..... Rigor and Franco though observed a superiority of the MZN stain above the trichrome stain ... of disease in diseased population hence its relevance in diagnosis, was noted to be high in trichrome and auramine stains.

  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. Enhanced

    Directory of Open Access Journals (Sweden)

    Martin I. Bayala

    2014-06-01

    Full Text Available Land Surface Temperature (LST is a key parameter in the energy balance model. However, the spatial resolution of the retrieved LST from sensors with high temporal resolution is not accurate enough to be used in local-scale studies. To explore the LST–Normalised Difference Vegetation Index relationship potential and obtain thermal images with high spatial resolution, six enhanced image sharpening techniques were assessed: the disaggregation procedure for radiometric surface temperatures (TsHARP, the Dry Edge Quadratic Function, the Difference of Edges (Ts∗DL and three models supported by the relationship of surface temperature and water stress of vegetation (Normalised Difference Water Index, Normalised Difference Infrared Index and Soil wetness index. Energy Balance Station data and in situ measurements were used to validate the enhanced LST images over a mixed agricultural landscape in the sub-humid Pampean Region of Argentina (PRA, during 2006–2010. Landsat Thematic Mapper (TM and Moderate Resolution Imaging Spectroradiometer (EOS-MODIS thermal datasets were assessed for different spatial resolutions (e.g., 960, 720 and 240 m and the performances were compared with global and local TsHARP procedures. Results suggest that the Ts∗DL technique is the most adequate for simulating LST to high spatial resolution over the heterogeneous landscape of a sub-humid region, showing an average root mean square error of less than 1 K.

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

    Science.gov (United States)

    Böthig, Antonia M; Hayn-Leichsenring, Gregor U

    2017-01-01

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

  17. Electrostatic control of the coffee stain effect

    Science.gov (United States)

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

    2013-11-01

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

  18. Erbium doped stain etched porous silicon

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez-Diaz, B. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Diaz-Herrera, B. [Departamento de Energia Fotovoltaica, Instituto Tecnologico de Energias Renovables (ITER), Poligono Industrial de Granadilla, 38611 S/C Tenerife (Spain); Guerrero-Lemus, R. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain)], E-mail: rglemus@ull.es; Mendez-Ramos, J.; Rodriguez, V.D. [Departamento de Fisica Fundamental, Experimental Electronica y Sistemas, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Hernandez-Rodriguez, C. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Martinez-Duart, J.M. [Departamento de Fisica Aplicada, C-XII, Universidad Autonoma de Madrid, 28049 Cantoblanco, Madrid (Spain)

    2008-01-15

    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{sub 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{sup 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.

  19. Microwave oven-based technique for immunofluorescent staining of paraffin-embedded tissues.

    Science.gov (United States)

    Long, Delwin J; Buggs, Colleen

    2008-02-01

    Immunohistochemical analysis of formalin-fixed paraffin-embedded tissues can be challenging due to potential modifications of protein structure by exposure to formalin. Heat-induced antigen retrieval techniques can reverse reactions between formalin and proteins that block antibody recognition. Interactions between antibodies and antigens are further enhanced by microwave irradiation, which has simplified immunohistochemical staining protocols. In this report, we modify a technique for antigen retrieval and immunofluorescent staining of formalin-fixed paraffin-embedded tissues by showing that it works well with several antibodies and buffers. This microwave-assisted method for antigen retrieval and immunofluorescent staining eliminates the need for blocking reagents and extended washes, which greatly simplifies the protocol allowing one to complete the analysis in less than 3 h.

  20. Deoxyribonuclease I (DNase I) typing from semen stains: low enzyme activity in vaginal fluids does not interfere with seminal DNase I typing from mixture stains.

    Science.gov (United States)

    Sawazaki, K; Yasuda, T; Nadano, D; Iida, R; Takeshita, H; Uchide, K; Kishi, K

    1993-09-01

    We describe the use of deoxyribonuclease I (DNase I) polymorphism for individualization of semen in body fluid stain mixtures, as a means of providing new and more useful information to practicing forensic biologists as a genetic marker. We have already reported that human DNase I isozyme patterns from different subjects are classifiable into ten groups. Isoelectric focusing of DNase I isozymes on polyacrylamide gel (IEF-PAGE, pH 3.5 to 5) was accomplished using a 0.5 mm thick gel. Pretreatment of semen samples with neuraminidase enhanced the isozyme band resolution and sensitivity. Activity detection using the dried agarose film overlay (DAFO) procedure was reliable, sensitive and simple, with high resolution, and the phenotypes of DNase I were determined in semen stains of about 0.3 microL stored at room temperature for up to a year in most of the samples tested. The DNase I types in semen stains were correlated with the types found in the corresponding blood and urine samples, although most of the vaginal fluid samples had no typable DNase I activity. This is considerably advantageous for seminal individualization from body fluid mixture stains in criminal cases. An evaluation of DNase I typing by IEF-PAGE and DAFO was also performed on casework samples submitted to our laboratory, and the results showed that DNase I was expected to be one of the most useful individualization marker of semen in practical application.

  1. Light microscopy with differential staining techniques for the characterisation and discrimination of insects versus marine arthropods processed animal proteins.

    Science.gov (United States)

    Ottoboni, Matteo; Tretola, Marco; Cheli, Federica; Marchis, Daniela; Veys, Pascal; Baeten, Vincent; Pinotti, Luciano

    2017-08-01

    The aim of this study was to evaluate the use of light microscopy with differential staining techniques for the discrimination of insect material from marine arthropods - classified as fishmeal. Specifically, three samples of single-species insect material, Hermetia illucens (HI), Bombyx mori (BM) and Tenebrio molitor (TM), and two samples of marine arthropods, shrimp material and krill, were analysed and compared after staining by two reagents to enhance fragment identification. Alizarin Red (AR) and Chlorazol Black (CB), which react respectively with calcium salts and chitin, were tested for their potential efficacy in distinguishing between insect and marine materials. Results indicated that AR failed to stain HI, BM and TM materials. By contrast, the three insect species materials tested were stained by CB. When shrimp fragments and krill were considered, AR and CB stained marine materials reddish-pink and light blue to black, respectively. By combining these results, it can be suggested that CB staining may efficiently be used to mark insect materials; AR does stain shrimp fragments but does not stain the tested insect material, indicating a possible approach for discriminating between insects and marine arthropods. However, since the present study was performed on pure materials and a small set of samples, possible implementation of this technique still needs to be confirmed in complex matrices such as compound feed.

  2. An automated double staining procedure for bone and cartilage.

    Science.gov (United States)

    Miller, D M; Tarpley, J

    1996-03-01

    Differential skeletal staining is an important part of developmental toxicologic studies. Traditionally these studies have required time-consuming differentiation of one or both stains used and careful attention to the maceration step to prevent specimen destruction. We present a fully automated protocol which does not require differentiation of either dye and incorporates a controlled maceration step which is highly reproducible. This has resulted in high quality staining that is reproducible, stable, and can be done in volume with minimal personnel time. The process involves the staining of skinned, eviscerated specimens fixed in 95% ethanol. Using an automated tissue processor, the specimen is stained in alcian blue for 24 hr, macerated in 3% potassium hydroxide for 24 hr and stained with murexide for 24 hr. The specimens are cleared and preserved in glycerol. Within three days specimens have red stained bone and blue stained cartilage. The procedure was developed using 20-day-old Sprague-Dawley rat fetuses to evaluate the feasibility of using the procedure for teratology studies involving the fetal skeleton. Evenly stained specimens can be examined within three days and stored for years without loss of staining.

  3. New Methylene Blue Stain for Malaria Detection on Thin Smears

    Directory of Open Access Journals (Sweden)

    Himanshu D. Mulay

    2017-01-01

    Full Text Available Background: Malaria is the most important parasitic infection of man. Microscopy remains gold standard in malaria diagnosis. Management of malaria requires rapid detection of parasite in human blood. Hence there is a need to develop another diagnostic method with less limitation, which will address this issue. Aim and Objectives: To find a low cost reliable and accurate method for malaria detection on peripheral smear. Material and Methods: A prospective study of 40 cases was done. Two thin smears were prepared for each case; one was stained with Leishman stain and other with new methylene blue stain and examined under oil immersion. The smears were examined individually by two pathologists and results were prepared. Different parasitic morphologic forms were looked for. Parasitemia percentage was calculated. We also compared number of fields required to diagnose with both stains in positive cases. Results: In this study we found that 25 (83.3% cases were detected in less than 50 fields using New Methylene blue stain against 18 (60.0% cases with Leishman stain. We also found 100% sensitivity and specificity for New Methylene blue stain, whereas Leishman stain showed 90% sensitivity and specificity of 85%. Conclusion: The detection of malaria parasite was considerably easy with New Methylene Blue stain and required less time in comparison with Leishman stain.

  4. [Application of histochemical staining in diagnosis of osteosarcomas].

    Science.gov (United States)

    Li, Qing; Gong, Xi-qi; Ma, Fu-cheng; Zhao, Yi-ling; Zhu, Xiao-hui

    2005-08-01

    To study the histochemical staining in the diagnosis of osteosarcoma. To compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification. With modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining. The modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.

  5. 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.

  6. 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.

  7. 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.

  8. Near-UV laser treatment of extrinsic dental enamel stains.

    Science.gov (United States)

    Schoenly, J E; Seka, W; Featherstone, J D B; Rechmann, P

    2012-04-01

    The selective ablation of extrinsic dental enamel stains using a 400-nm laser is evaluated at several fluences for completely removing stains with minimal damage to the underlying enamel. A frequency-doubled Ti:sapphire laser (400-nm wavelength, 60-nanosecond pulse duration, 10-Hz repetition rate) was used to treat 10 extracted human teeth with extrinsic enamel staining. Each tooth was irradiated perpendicular to the surface in a back-and-forth motion over a 1-mm length using an ∼300-µm-diam 10th-order super-Gaussian beam with fluences ranging from 0.8 to 6.4 J/cm(2) . Laser triangulation determined stain depth and volume removed by measuring 3D surface images before and after irradiation. Scanning electron microscopy evaluated the surface roughness of enamel following stain removal. Fluorescence spectroscopy measured spectra of unbleached and photobleached stains in the spectral range of 600-800 nm. Extrinsic enamel stains are removed with laser fluences between 0.8 and 6.4 J/cm(2) . Stains removed on sound enamel leave behind a smooth enamel surface. Stain removal in areas with signs of earlier cariogenic acid attacks resulted in isolated and randomly located laser-induced, 50-µm-diam enamel pits. These pits contain 0.5-µm diam, smooth craters indicative of heat transfer from the stain to the enamel and subsequent melting and water droplet ejection. Ablation stalling of enamel stains is typically observed at low fluences (Laser ablation of extrinsic enamel stains at 400 nm is observed to be most efficient above 3 J/cm(2) with minimal damage to the underlying enamel. Unsound underlying enamel is also observed to be selectively removed after irradiation. Copyright © 2012 Wiley Periodicals, Inc.

  9. 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

  10. 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...... sources makes it difficult to decide which equipment is the best for treating port-wine stains....

  11. 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...

  12. 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.

  13. 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

  14. Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components.

    Science.gov (United States)

    Belaldavar, C; Hallikerimath, S; Angadi, P V; Kale, A D

    2014-11-01

    The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues.

  15. 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...

  16. Christendom's Narratives and the Stained Glass Designs of Yusuf ...

    African Journals Online (AJOL)

    Stained glass paintings, the most distinctive accessory in the interior repertoire of Christian ecclesiastic spaces, is the subject matter of this study. This paper attempts a recast of Christendom's narratives in the stained glass designs of Yusuf Cameron Adebayo Grillo as the distinctive overarching mechanism of the ...

  17. Standardization in biological staining. The influence of dye manufacturing

    DEFF Research Database (Denmark)

    Lyon, H

    2000-01-01

    for biological staining, standardization of all procedures and reagents is mandatory. In this paper, I focus particularly on dyes and consider the possibilities for obtaining standardized dyes. In general practice, most biological staining takes place with available commercial dyes. These dyes may or may...

  18. Diagnostic utility of the genital Gram stain in ED patients.

    Science.gov (United States)

    Stefanski, Peter; Hafner, John W; Riley, Shanda L; Sunga, Kharmene L Y; Schaefer, Timothy J

    2010-01-01

    The study aimed to determine the diagnostic usefulness of the genital Gram stain in an emergency department (ED) population. A linked-query of an urban, tertiary-care, university- affiliated hospital laboratory database was conducted for all completed Chlamydia trachomatis and Neisseria gonorrhoeae DNA probes, Trichomonas vaginalis wet preps, and genital Gram stains performed on ED patient visits between January and December 2004. Positive criteria for a Gram stain included greater than 10 white blood cells per high-power field, gram-negative intracellular/extracellular diplococci (suggesting N gonorrhoeae), clue cells (suggesting T vaginalis), or direct visualization of T vaginalis organisms. DNA probes were used as the gold standard definition for N gonorrhoeae and C trachomatis infection. Of 1511 initially eligible ED visits, 941 were analyzed (genital Gram stain and DNA probe results both present), with a prevalence of either C trachomatis or N gonorrhoeae of 11.4%. A positive genital Gram stain was 75.7% sensitive and 43.3% specific in diagnosing either C trachomatis and/or N gonorrhoeae infection, and 80.4% sensitive and 32.2% specific when the positive cutoff was lowered to more than 5 white blood cells/high-power field. No Gram stains were positive for T vaginalis (with 47 positive wet mounts), and clue cells were noted on 117 Gram stains (11.6%). Gram stains in isolation lack sufficient diagnostic ability to detect either C trachomatis or N gonorrhoeae infection in the ED.

  19. 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 ...

  20. 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 ...

  1. [Lugol staining for esophageal carcinoma and influence of radiotherapy].

    Science.gov (United States)

    Miyamoto, H; Adachi, W; Koike, S; Koide, N; Iida, F

    1993-01-01

    For evaluating the endoscopic staining of the esophageal carcinoma with lugol solution, 50 patients who underwent esophagectomy for carcinoma were subjected to this study. Among the 50 patients, 21 were radiated before surgery and 29 were not radiated. 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 non-radiation group, but both areas of 28.6% cases disagreed in radiation group. On the second step, 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-radiation group and 71.4% in the radiation group. As one of the causes of the large non-corresponding rate in the radiation group, radiation esophagitis was demonstrated. It can be finally concluded that the reliability of endoscopic lugol staining is reduced by preoperative irradiation.

  2. Are port wines stains a feature of tuberous sclerosis?

    Science.gov (United States)

    Ben-Amitai, D; Halachmi, S; Lapidoth, M

    2011-07-01

    Tuberous sclerosis complex is a multisystem inherited disorder characterized by the development of tumour-like growths in brain, skin and other organs. Although cutaneous vascular anomalies are not considered a common manifestation, we have encountered co-occurrence of port wine stains and tuberous sclerosis. To assess the prevalence of port wine stain in patients with previously diagnosed tuberous sclerosis. All cases diagnosed with tuberous sclerosis at two tertiary care centres from 2000 to 2009 were reviewed. Cases with clinically documented port wine stains were included for evaluation. Of 24 patients diagnosed with tuberous sclerosis, three (12.5%) had clinically evident port wine stains. The prevalence of port wine stains in this series of tuberous sclerosis patients was significantly higher than the 0.3% prevalence of port wine stain in the general population. Port wine stain rate in this population was significantly greater than the expected rate. Further studies are needed to assess the frequency of port wine stains in tuberous sclerosis and to clarify whether the finding should be added to the list of cutaneous features of tuberous sclerosis. © 2010 The Authors. Journal of the European Academy of Dermatology and Venereology © 2010 European Academy of Dermatology and Venereology.

  3. THIONIN STAINING OF PARAFFIN AND PLASTIC EMBEDDED SECTIONS OF CARTILAGE

    NARCIS (Netherlands)

    BULSTRA, SK; DRUKKER, J; KUIJER, R; BUURMAN, WA; VANDERLINDEN, AJ

    The usefulness of thionin for staining cartilage sections embedded in glycol methacrylate (GMA) and the effect of decalcification on cartilage sections embedded in paraffin and GMA were assessed. Short decalcification periods using 5% formic acid or 10% EDTA did not influence the staining properties

  4. Mercury-selenium interactions in relation to histochemical staining of mercury in the rat liver

    DEFF Research Database (Denmark)

    Baatrup, E; Thorlacius-Ussing, O; Nielsen, H L

    1989-01-01

    micrograms of Se g-1 body weight as sodium [75Se]selenite. All the rats were killed 1 h later. Homogenized liver samples were prepared for mercury analysis by two different methods: alkaline digestion and ultrasonic disintegration. Quantitative chemical analysis based on benzene extraction......Selenium has been suggested to enhance the histochemical staining of mercury when sections of tissue are subjected to the silver-enhancement method. In the present study, histochemical staining patterns of mercury in tissue sections of rat livers were compared with the actual content of organic...... and inorganic Hg in the livers, in both the presence and the absence of Se. Rats were injected intravenously with 5 micrograms of Hg g-1 body weight as methyl [203Hg] mercury chloride (MeHg) or as [203Hg]mercuric chloride (Hg2+). After 2 h, half the rats received an additional intraperitoneal injection of 2...

  5. 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.

  6. Homogeneous luminescent stain etched porous silicon elaborated by a new multi-step stain etching method

    Energy Technology Data Exchange (ETDEWEB)

    Hajji, M., E-mail: mhajji2001@yahoo.fr [Laboratoire de Photovoltaïque, Centre de Recherche et des Technologies de l’Energie, Technopôle de Borj-Cédria BP 95, Hammam-Lif 2050 (Tunisia); Institut Supérieur d’Electronique et de Communication de Sfax, route Menzel Chaker Km 0.5, BP 868, Sfax 3018 (Tunisia); Khalifa, M.; Slama, S. Ben; Ezzaouia, H. [Laboratoire de Photovoltaïque, Centre de Recherche et des Technologies de l’Energie, Technopôle de Borj-Cédria BP 95, Hammam-Lif 2050 (Tunisia)

    2013-11-01

    This paper presents a new method to produce porous silicon which derived from the conventional stain etching (SE) method. But instead of one etching step that leads to formation of porous layer, the substrate is subjected to an initial etching step with a duration Δt{sub 0} followed by a number of supplementary short steps that differs from a layer to another. The duration of the initial step is just the necessary time to have a homogenous porous layer on the whole surface of the substrate. It was found that this duration is largely dependent of the doping type and level of the silicon substrate. The duration of supplementary steps was kept as short as possible to prevent the formation of bubbles on the silicon surface during silicon dissolution which leads generally to inhomogeneous porous layers. It is found from surface investigation by atomic force microscopy (AFM) that multistep stain etching (MS-SE) method allows to produce homogeneous porous silicon nanostructures compared to the conventional SE method. The chemical composition of the obtained porous layers has been evaluated using Fourier transform infrared spectroscopy (FTIR). Photoluminescence (PL) measurement shows that porous layers produced by SE and MS-SE methods have comparable spectra indicating that those layers are composed of nanocrystallites with comparable sizes. But the intensity of photoluminescence of layer elaborated by MS-SE method is higher than that elaborated by the SE method. Total reflectance characteristics show that the presented method allows the production of porous silicon layers with controllable thicknesses and optical properties. Results for porous silicon layers elaborated on heavily doped n-type silicon show that the reflectance can be reduced to values less than 3% in the major part of the spectrum.

  7. 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

  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. 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.

  10. 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.

  11. Visualising the 3D microstructure of stained and native intervertebral discs using X-ray microtomography.

    Science.gov (United States)

    Disney, C M; Madi, K; Bodey, A J; Lee, P D; Hoyland, J A; Sherratt, M J

    2017-11-24

    Intervertebral disc degeneration (IVDD) is linked to low back pain. Microstructural changes during degeneration have previously been imaged using 2D sectioning techniques and 3D methods which are limited to small specimens and prone to inducing artefacts from sample preparation. This study explores micro computed X-ray tomography (microCT) methods with the aim of resolving IVD 3D microstructure whilst minimising sample preparation artefacts. Low X-ray absorption contrast in non-mineralised tissue can be enhanced using staining and phase contrast techniques. A step-wise approach, including comparing three stains, was used to develop microCT for bovine tail IVD using laboratory and synchrotron sources. Staining successfully contrasted collagenous structures; however not all regions were stained and the procedure induced macroscopic structural changes. Phase contrast microCT of chemically fixed yet unstained samples resolved the nucleus pulposus, annulus fibrosus and constituent lamellae, and finer structures including collagen bundles and cross-bridges. Using the same imaging methods native tissue scans were of slightly lower contrast but free from sample processing artefacts. In the future these methods may be used to characterise structural remodelling in soft (non-calcified) tissues and to conduct in situ studies of native loaded tissues and constructs to characterise their 3D mechanical properties.

  12. An in vitro screening assay for dental stain cleaning.

    Science.gov (United States)

    Wang, Changxiang; Lucas, Robert; Smith, Anthony J; Cooper, Paul R

    2017-01-09

    The present study aimed to develop an in vitro model for stain removal from natural enamel for the assessment and comparison of oral hygiene products. Bovine teeth (n = 8 per group) were ground/polished to provide flat enamel specimens and ferric-tannate deposits were precipitated onto the enamel surfaces. The ferric-tannate stained enamel specimens were brushed using an in vitro tooth-brushing simulator with slurries containing commercially available toothpaste products, dental abrasive particles, and sodium tripolyphosphate (STP) solutions of different concentrations. The colour of the enamel surfaces was measured using a spectrophotometer before and after stain application as well as after the brushing treatments. Differences in stain removal efficacy were found between the toothpastes categorised as whitening and non-whitening comprising of different types of dental abrasives (hydrated silica and alumina). A mean value of 27% for stain removal was detected for the three non-whitening toothpastes and 59% of stain removal was detected for the three whitening toothpastes after 1000 strokes. Compared with the slurry with Zeodent 113 abrasive alone, the addition of STP provided better performance for stain removal under the same brushing conditions (mean value of 62% for Zeodent 113 abrasive alone and 72% with the addition of 5% (w/w) STP after 1000 strokes). No difference was evident between the STP concentration of 5% (w/w) and 10% (w/w). The ferric-tannate/bovine enamel model reported here provides good stain retention, is rapidly and easily prepared, and is shown to be progressively and reproducibly sensitive to toothbrushing using different toothpastes and surfactant/chelating agent solutions. Importantly, it provides good discrimination between various oral hygiene products. The stain removal assay reported here has considerable potential to enable comparative assessments of different toothpaste types in terms of their cleaning capabilities.

  13. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    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......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...

  14. News from the Biological Stain Commission No. 11

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2012-01-01

    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......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...

  15. Clinical studies to determine the effectiveness of a whitening toothpaste at reducing stain (using a forced stain model).

    Science.gov (United States)

    Moran, J; Claydon, N C A; Addy, M; Newcombe, R

    2005-02-01

    Two single centre, randomized single-blind, crossover studies were performed, to compare the effect of a test toothpaste with a conventional fluoride paste in the inhibition and removal of extrinsic dental stain promoted by repeated chlorhexidine/tea rinses. These studies used 24 subjects in each of two separate clinical trials. On the Friday before each trial period, the subjects received a prophylaxis to remove all staining, plaque and calculus deposits. On the following Monday, subjects were checked whether they were stain free and then under direct supervision they rinsed with a 0.2% chlorhexidine mouthrinse, immediately followed by a rinse with a warm black tea solution. This cycle was repeated hourly eight times throughout the day and on the following days until the Friday. In addition subjects also received daily a single toothpaste slurry rinse or control water rinse in the morning and lunchtime. No other form of oral hygiene was permitted during this period. On the Friday, both stain area and intensity was assessed using the Lobene Stain Index. For the stain removal study, stain was promoted again using chlorhexidine and tea rinses. After 4 days, stain was measured both prior to and immediately after brushing with the allocated toothpaste for 2 min. Subjects were then instructed to use the toothbrush at home according to their normal oral hygiene practices. On the following Wednesday, the amount of stain present was re-assessed. Each subject subsequently received a thorough prophylaxis to remove all plaque calculus and staining before starting the following periods of the study. The study showed no difference in the ability of the test whitening toothpaste, control toothpaste and water control at inhibiting stain. There was also only a small difference (3.5% for product of area and intensity) between the ability of the two toothpastes to help remove stain after a single brushing. The difference was however in favour of the test product which approached a

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

    African Journals Online (AJOL)

    Rukevwe S. Abraka

    2016-09-14

    Sep 14, 2016 ... After fixation of specimens, the cut- up was done, specimens were put in cassettes ... Rinse in stock solution of acetic acid. 2. Stain in giemsa .... analysis of tendon collagen using two-dimensional polarized light microscopy.

  17. Dye purity and dye standardization for biological staining

    DEFF Research Database (Denmark)

    Lyon, H O

    2002-01-01

    This review starts with a short discussion of what is meant by a pure dye and a brief description of how dyes are produced. A listing of the types of impurities encountered in dyes is made, followed by technical investigations and assessments of dyes and their impurities including methods...... 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....

  18. Spectral analysis of blood stains at the crime scene

    NARCIS (Netherlands)

    Edelman, G.J.

    2014-01-01

    In this thesis, we propose the use of several optical techniques for the detection, identification, and age estimation of blood stains. We explore the visible, near infrared, and mid infrared wavelength range for this purpose.

  19. 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.

  20. News from the Biological Stain Commission, No. 17

    DEFF Research Database (Denmark)

    Lyon, H O

    2016-01-01

    In the 17(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 20(th) meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test...... systems held on October 15 - 17, 2014 in Toronto, Canada, and from the 29(th) meeting of CEN/TC 140 In vitro diagnostic medical devices held on February 3, 2015 in Berlin, Germany....

  1. Immunohistochemical CD3 staining detects additional patients with celiac disease.

    Science.gov (United States)

    Mubarak, Amani; Wolters, Victorien M; Houwen, Roderick H J; ten Kate, Fiebo J W

    2015-06-28

    To investigate whether performing immunohistochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease. Biopsies obtained from 159 children were stained by hematoxylin and eosin (HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently. Differences in evaluation between the routine HE sections and CD3 staining were present in 20 (12.6%) cases. In 10 (6.3%) patients the diagnosis of celiac disease (Marsh II and III) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10 (12.3%) patients, the detection of sole intra-epithelial lymphocytosis (Marsh I) improved. Nine patients were found to have Marsh I on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh I was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections. Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.

  2. 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....

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

    Science.gov (United States)

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

    2014-09-01

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

  4. MANAJEMEN SARANA DAN PRASARANA PENDIDIKAN DI STAIN PAMEKASAN

    Directory of Open Access Journals (Sweden)

    M. Muchlis Solichin

    2011-07-01

    Full Text Available Mediums management and pre-mediums represent an absolute done in an higher education institute, because Mediums and premediums in education management represent the absolut condition in the effort to reach the target which is expected. Thereby, Every the education organizer have to pay attention and conscripting the mind and energy to carry out education management that is professional and fulfill Standard National Education ( SNP. This Research copes to comprehend the mediums and pre-mediums management of education in STAIN Pamekasan, because during this time of mediums and basic mediums management are not yet showing its idealitas. This research is focussed at; a How mediums and pre-mediums menegement in STAIN Pamekasan ?,and b what Factors influencing mediums and pre-mediums management in STAIN Pamekasan ?. This research uses the qualitative type by using observation, interview, and documentation method. Based the rearch done, to be expressed that the first of STAIN Pamekasan conduct mediums and pre-mediums manegement still have the centralization character of top down, either in the case of planning, organizational, observation, and assessment of mediums and pre-mediums management owned, second in some cases of STAIN Pamekasan do not yet manage the mediums and pre-mediums management because they are caused by factor is its lack of management professionalism, either when doing the planning, organizational, treatment and observation or evaluation. Based the matter above, hence, suggested that STAIN Pamekasan carry out the mediums and pre-mediums management of education professionally.

  5. Lugol staining pattern and histology of esophageal lesions.

    Science.gov (United States)

    Mori, M; Adachi, Y; Matsushima, T; Matsuda, H; Kuwano, H; Sugimachi, K

    1993-05-01

    To analyze the relationship between Lugol unstained areas and their histologic features, we applied the Lugol test to 24 specimens of resected esophagus. The staining patterns were graded into four types: grade I, hyperstaining; grade II, normal greenish brown staining; grade III, less intense staining; and grade IV, unstained. Most of the grade IV lesions were invasive carcinomas, carcinomas in situ, or severe dysplasia. The carcinomas in situ and the intraepithelial extension of the carcinomas, which were difficult to detect, were clearly shown as grade IV. On the other hand, moderate to mild dysplasia or atrophy showed grade III staining. Grade IV lesions showed well-demarcated sharp margins, whereas grade III lesions showed ill-demarcated dull margins. The grade III carcinomas, however, by the Lugol test, showed well-demarcated margins. Histologic evaluation disclosed that the staining intensity reflected well the thickness of the glycogen-containing cell layer in the lesion. The sharpness of the margin reflected the abrupt or gradual change from the glycogen-containing to non-containing cell layers. These findings suggest 1) the usefulness of the staining pattern of the Lugol test for the diagnosis of esophageal lesions such as squamous cell carcinoma and severe dysplasia, and 2) the usefulness of the Lugol test for precise delineation of the proximal resection line during surgery of esophageal carcinomas with unexpected wide extension.

  6. Effect of finishing and polishing on the color stability of a composite resin immersed in staining solutions

    Directory of Open Access Journals (Sweden)

    Maiara Justo Polli

    2015-01-01

    Full Text Available Objective: To evaluate the influence of finishing/polishing methods and staining solutions using different immersion periods on the color stability of a microhybrid composite resin. Materials and Methods: Ninety specimens were fabricated using a stainless steel mold and polyester strips. The samples were randomly divided into five groups according to the finishing and polishing performed: Control group (no surface treatment was performed, Diamond Pro group, Diamond burs group, Enhance group, and SiC paper group. After finishing and polishing, six samples from each group were immersed in coffee, red wine, or water for 30 days. The color measurements were obtained using digital photography before immersion and after 7, 15, and 30 days of immersion. The red, green, and blue values provided by the Adobe Photoshop software were converted into CIELab values. A three-way analysis of variance and Tukey's test were used for statistical analysis (P ≤ 0.05. Results: The finishing and polishing methods, staining solutions, immersion times, and their interaction had statistically significant effects on the color change (P = 0.00. Coffee and red wine caused intense staining. Among the polishing methods, the highest color change value was observed in the control group (P < 0.05 and the Diamond Pro disks provided the most stain-resistant surfaces (P ≤ 0.05. Conclusion: The finishing and polishing method, staining solution, and immersion time influences the color stability. Finishing and polishing should be applied to obtain a more stain-resistant surface.

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

    Science.gov (United States)

    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.

  8. 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 (pstaining 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

  9. Clinical Study to Assess the Stain Removal Effectiveness of a Tooth Whitening Regimen with Added Whitening Booster.

    Science.gov (United States)

    Ghassemi, A; Hooper, W; Vorwerk, L; Patel, V; Sheth, J

    2015-01-01

    This randomized, controlled clinical trial was conducted to assess the extrinsic stain reduction achieved by brushing with a whitening dentifrice and powered toothbrush, and to determine whether the addition of a whitening booster paste to this regimen would enhance its stain reducing effectiveness. Sixty qualifying subjects were randomly assigned either to Regimen One, a whitening dentifrice (Arm & Hammer Truly Radiant [TR] toothpaste] and powered toothbrush (Arm & Hammer Truly Radiant [TR] Extra Whitening Spinbrush); Regimen Two, the dentifrice and powered toothbrush with the addition of a whitening booster; or Regimen Three, a negative control (Colgate Cavity Protection toothpaste and an ADA standard manual brush). They were instructed in the use of their assigned products and then brushed unsupervised at home for two minutes, twice daily, for 14 days. Extrinsic tooth stain was assessed at baseline and after two, five, and 14 days using a Modified Lobene Stain Index (MLSI) with Lobene inclusion criteria of ≥ 1.5. All three treatment groups had statistically significant (p toothpaste and Truly Radiant Spinbrush provides safe and effective stain removal that can be further enhanced by the addition of the whitening booster.

  10. CD3 immunohistochemical staining in diagnosis of lymphocytic colitis.

    Science.gov (United States)

    Fiehn, Anne-Marie Kanstrup; Engel, Ulla; Holck, Susanne; Munck, Lars Kristian; Engel, Peter Johan Heiberg

    2016-02-01

    Microscopic colitis (MC) is a common cause of chronic watery diarrhea. Traditionally, MC encompasses the 2 subgroups lymphocytic colitis (LC) and collagenous colitis, but recently, an additional subgroup, MC incomplete, has been introduced. Distinguishing between the subgroups relies exclusively on histopathologic evaluation. In the present study, 4 pathologists evaluated 156 archived biopsies originally diagnosed as LC or LC incomplete (LCi). Each pathologist assigned a diagnosis of LC, LCi, or nonspecific inflammation to all cases at 2 independent assessments. At the first assessment, hematoxylin and eosin (HE) stainings were available. At the second assessment, a supplementary CD3 immunohistochemical staining was also available. The aim was to evaluate whether a supplementary CD3 would increase the diagnostic agreement among pathologists, and whether a CD3 stain would change the diagnosis based on HE staining only. After the complete assessment, the cases were divided into 3 groups, that is, full agreement, partial agreement, and disagreement. The CD3 staining increased the number of cases with full agreement from 60 to 78. One hundred thirty-one cases with agreement or partial diagnostic agreement based on HE + CD3 were compared with the HE diagnoses. In 44 (34%) of 131 cases, CD3 changed the diagnosis. Cases assigned to the LCi category based on HE were often changed by a supplementary CD3. Conclusively, it is recommended to use a CD3 before giving the histopathologic diagnosis of LCi. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Identification and quantification of microplastics using Nile Red staining.

    Science.gov (United States)

    Shim, Won Joon; Song, Young Kyoung; Hong, Sang Hee; Jang, Mi

    2016-12-15

    We investigated the applicability of Nile Red (NR), a fluorescent dye, for microplastic analysis, and determined the optimal staining conditions. Five mg/L NR solution in n-hexane effectively stained plastics, and they were easily recognized in green fluorescence. The NR staining method was successfully applied to micro-sized polyethylene, polypropylene, polystyrene, polycarbonate, polyurethane, and poly(ethylene-vinyl acetate), except for polyvinylchloride, polyamide and polyester. The recovery rate of polyethylene (100-300μm) spiked to pretreated natural sand was 98% in the NR stating method, which was not significantly (p<0.05) different with FT-IR identification. The NR staining method was suitable for discriminating fragmented polypropylene particles from large numbers of sand particles in laboratory weathering test samples. The method is straightforward and quick for identifying and quantifying polymer particles in the laboratory controlled samples. Further studies, however, are necessary to investigate the application of NR staining to field samples with organic remnants. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Usefulness of the stool Wright's stain in the emergency department.

    Science.gov (United States)

    DuBois, D; Binder, L; Nelson, B

    1988-01-01

    A prospective study was conducted to determine if a Wright's stain of stool specimen to detect fecal leukocytes was accurate in predicting the presence of a bacterial pathogen on stool culture. Entry criteria were patient age greater than or equal to 3 months and diarrhea of greater than 1 day. The patient population was drawn from an urban county hospital emergency department on the Texas-Mexican border. A total of 69 patients were evaluated by both routine stool culture and stool Wright's stain. Twenty-three were evaluated for parasitic pathogens. There were seventeen cultures positive for bacterial pathogens and twenty-three positive Wright's stains. Bacterial isolates included Shigella, Salmonella and Campylobacter. Also detected were Giardia, Shistosoma, Blastocytis and Cryptosporidium. The sensitivity of a Wright's stain positive for fecal leukocytes for the presence of a bacterial pathogen by culture was 82%, with a specificity of 83%. These were significantly correlated with a positive culture for a bacterial pathogen (P less than .01). The predictive value of a positive result was 61%, and predictive value of a negative result was 94%, for bacterial pathogens. The Wright's stain is a useful tool for the early presumptive diagnosis of infectious bacterial diarrhea in the emergency department.

  13. When one plus one equals more than two - a novel stain for renal biopsies is a combination of two classical stains

    OpenAIRE

    Brodsky, Sergey V.; Albaward, Alia; Satoskar, Anjali A.; Nadasdy, Gyongyi; Nadasdy, Tibor

    2010-01-01

    Histologic evaluation of renal biopsies includes multiple ancillary stains, including Periodic acid-Schiff ’s (PAS) and Masson’s trichrome (Trichrome). Herein we report an innovative doublestain, derived from two standard stains (PAS and Trichrome). This novel stain not only has advantages of both ancestor stains, but became more distinguishable and colorful, when basement membranes stain darkviolet, whereas the interstitial collagen remains blue. This allows the pa...

  14. MEGARA Optics: stain removal in PBM2Y prisms

    Science.gov (United States)

    Aguirre-Aguirre, D.; Izazaga-Pérez, R.; Villalobos-Mendoza, B.; Carrasco, E.; Gil de Paz, A.; Gallego, J.; Iglesias, J.

    2017-01-01

    MEGARA is the new integral-field and multi-object optical spectrograph for the GTC. For medium and high resolution, the dispersive elements are volume phase holographic gratings, sandwiched between two flat windows and two prisms of high optical precision. The prisms are made of Ohara PBM2Y optical glass. After the prisms polishing process, some stains appeared on the surfaces. For this, in this work is shown the comparative study of five different products (muriatic acid, paint remover, sodium hydroxide, aqua regia and rare earth liquid polish) used for trying to eliminate the stains of the HR MEGARA prisms. It was found that by polishing with the hands the affected area, and using a towel like a kind of pad, and polish during five minutes using rare earth, the stains disappear completely affecting only a 5% the rms of the surface quality. Not so the use of the other products that did not show any apparent result.

  15. A Staining Protocol for Identifying Secondary Compounds in Myrtaceae

    Directory of Open Access Journals (Sweden)

    Hernan A. Retamales

    2014-10-01

    Full Text Available 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.

  16. 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

  17. 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.

  18. 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.

  19. Positive staining for cellulose in oral pulse granuloma

    DEFF Research Database (Denmark)

    Virkkunen, Sirke; Wolff, Henrik; Haglund, Caj

    2017-01-01

    the hyaline rings contain cellulose. Study Design: Using a newly developed staining method for cellulose, we studied 18 histologic samples diagnosed as OPG, in addition to 3 samples originally diagnosed as "normal" foreign body reactions. In our study, visualization of cellulose is based on its specific...... binding to the carbohydrate binding module of β-1,4-glycanase. Results: All samples diagnosed as OPG were positive for cellulose staining localized in hyaline rings. In addition, 1 lesion (of 3), first diagnosed as a foreign body reaction without the presence of hyaline rings, was positive for cellulose...... by horseradish peroxidase staining. Conclusions: We show for the first time that cellulose is present in OPG lesions, indicating that cellulose might be the initial cause of formation of these lesions....

  20. Extrinsic Stain Removal Effectiveness of a New Whitening Dentifrice.

    Science.gov (United States)

    Ghassemi, A; Vorwerk, L; Hooper, W; Cirigliano, A; DeSciscio, P; Nathoo, S

    2015-01-01

    This study was conducted to evaluate the effectiveness of Arm & Hammer (A&H) Truly Radiant Rejuvenating toothpaste in removing extrinsic tooth stain compared to that of a conventional fluoride/silica-containing dentifrice. This was a randomized, examiner-blind, parallel-design study with two groups of subjects who brushed unsupervised with their assigned dentifrice for two minutes, twice daily, for five days. Extrinsic stain was measured on the labial surfaces of the eight incisor teeth by the Modified Lobene Stain Index (MLSI) at baseline and following five days of product use. After balancing for baseline MLSI, beverage and tobacco use, fifty-four healthy adults with existing stain were randomly distributed into two comparable groups: Arm and Hammer Truly Radiant Rejuvenating toothpaste or Colgate Cavity Protection toothpaste (negative control). Within-treatment comparisons between baseline and day five were made using matched-pair t-tests, and between-treatment comparisons of MSLI scores were performed using ANCOVA, with baseline scores as covariates. Twenty-eight subjects in the Truly Radiant Rejuvenating toothpaste group and twenty-six subjects in the negative control group completed the study. The groups had comparable mean scores at baseline (p > 0.05). The Truly Radiant Rejuvenating toothpaste produced a statistically significant 23.1% total (composite) stain reduction from baseline after five days of product use (p 0.05). Between-treatment analysis showed statistically significantly (p toothpaste compared to the Colgate control following five days of product use. There were no adverse events reported during the study. The A&H Truly Radiant Rejuvenating toothpaste is safe and effective in reducing extrinsic stain compared to a regular toothpaste control.

  1. Chemical aspects of santalin as a histological stain.

    Science.gov (United States)

    Banerjee, A; Mukherjee, A K

    1981-03-01

    Recent research on the chemical nature of the red dyes isolated from Pterocarpus santalinus and certain West African plants, viz., Baphia nitida, Pterocarpus osun and Pterocarpus soyauxii, have been reviewed. P. santalinus contains santalins A, B and C, but no santarubin. Santalins and santarubins have been found in P. osun, P. soyauxii and B. nitida. The structural formulae of the santalins are presented and their differences from santarubins indicated. Santalins A and B have some similarities in structure with hematein. This is probably responsible for their staining properties; the possible mechanism of staining is discussed.

  2. DETECTION OF TISSUE MYCOBACTERIUM TUBERCULOSIS BY DIFFERENTIATING IMMUNOPEROXIDASE STAINING

    Directory of Open Access Journals (Sweden)

    A. P. Lysenko

    2014-01-01

    Full Text Available Staining impression smears from organ and tissues with peroxidase conjugated antibodies to Mycobacterium tuberculosis complex antigens, followed by visualization with diaminobenzidine and Kinyoun stains, ensured the painting of acid-resistant Mycobacterium tuberculosis forms to rubin red, acid-susceptible ones to brown, and tissue cells and microorganisms of other species to blue. Typical bacilli were absent in the lymph nodes of patients and animals with latent infection, but acid-resistant (rubin-red granular forms were encountered in the granulomatous masses. Brown fat cells containing mycobacterial antigens, as well as acid-susceptible granular, reticular, fungoid, and rod-like forms were also found in considerable quantities.

  3. Influence of stain etching on low minority carrier lifetime areas of multicrystalline silicon for solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Montesdeoca-Santana, A. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38206 La Laguna (Spain); Fraunhofer Institute for Solar Energy Systems, Laboratory and Servicecenter Gelsenkirchen, Auf der Reihe 2, 45884 Gelsenkirchen (Germany); Gonzalez-Diaz, B. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38206 La Laguna (Spain); Departamento de Energia Fotovoltaica, Instituto Tecnologico y de Energias Renovables. Poligono Industrial de Granadilla s/n, 38600 San Isidro-Granadilla de Abona (Spain); Jimenez-Rodriguez, E. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38206 La Laguna (Spain); Ziegler, J. [Fraunhofer Institute for Solar Energy Systems, Laboratory- and Servicecenter Gelsenkirchen. Auf der Reihe 2, 45884 Gelsenkirchen (Germany); Velazquez, J.J. [Departamento de Fisica Fundamental y Experimental, Electronica y Sistemas, Universidad de La Laguna. Avda. Astrofisico Francisco Sanchez, 38206 La Laguna (Spain); Hohage, S.; Borchert, D. [Fraunhofer Institute for Solar Energy Systems, Laboratory and Servicecenter Gelsenkirchen. Auf der Reihe 2, 45884 Gelsenkirchen (Germany); Guerrero-Lemus, R., E-mail: rglemus@ull.es [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38206 La Laguna (Spain)

    2011-11-15

    Highlights: > An enhanced minority carrier lifetime at extended defects in multicrystalline silicon is observed with the use of HF/HNO{sub 3} stain etching to texture the surface. > FTIR analysis shows no influence of oxide passivation in this effect. > SEM images show a preferential etching at extended defects suggesting smoothing at defects as one of the causes for the reduced recombination activity. > LBIC images show a reduction in IQE at extended defects in HF/HNO{sub 3} textured multicrystalline solar cells. - Abstract: In this work the use of HF/HNO{sub 3} solutions for texturing silicon-based solar cell substrates by stain etching and the influence of texturing on minority carrier lifetimes are studied. Stain etching is currently used to decrease the reflectance and, subsequently improve the photogenerated current of the cells, but also produces nanostructures on the silicon surface. In the textured samples it has been observed that an improvement on the minority carrier lifetime with respect to the samples treated with a conventional saw damage etching process is produced on grain boundaries and defects, and the origin of this effect has been discussed.

  4. Effects of the Gram stain on microspheres from thermal polyamino acids.

    Science.gov (United States)

    FOX, S W; YUYAMA, S

    1963-02-01

    Fox, Sidney W. (The Florida State University, Tallahassee) and Shuhei Yuyama. Effects of the Gram stain on microspheres from thermal polyamino acids. J. Bacteriol. 85:279-283. 1963.-Microspheres produced from acid proteinoid accept the Gram stain. The stain is negative, but microspheres produced from mixtures containing a sufficient proportion of lysine proteinoid stain positive. Microspheres produced from mixtures containing the appropriate proportions contain individuals which stain positive and others which stain negative.

  5. EFFECTS OF THE GRAM STAIN ON MICROSPHERES FROM THERMAL POLYAMINO ACIDS1

    Science.gov (United States)

    Fox, Sidney W.; Yuyama, Shuhei

    1963-01-01

    Fox, Sidney W. (The Florida State University, Tallahassee) and Shuhei Yuyama. Effects of the Gram stain on microspheres from thermal polyamino acids. J. Bacteriol. 85:279–283. 1963.—Microspheres produced from acid proteinoid accept the Gram stain. The stain is negative, but microspheres produced from mixtures containing a sufficient proportion of lysine proteinoid stain positive. Microspheres produced from mixtures containing the appropriate proportions contain individuals which stain positive and others which stain negative. Images PMID:13959050

  6. Crystal violet stain as a selective stain for the assessment of mitotic figures in oral epithelial dysplasia and oral squamous cell carcinoma.

    Science.gov (United States)

    Jadhav, Kiran B; Ahmed Mujib, B R; Gupta, Nidhi

    2012-01-01

    Assessment of mitotic figures (MFs) is routinely practiced as prognostic indicator in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC), but identification of MFs poses a problem in terms of staining characteristics. To evaluate effectiveness of crystal violet stain for staining of MFs and its comparison with hematoxylin and eosin (H and E) stain. Study sample includes archival tissues embedded in paraffin blocks diagnosed as OED (n = 30) and OSCC (n = 30). The control group comprised of tissue specimen from oral mucosa of healthy volunteers (n = 30). Two serial sections of each tissue specimen were stained separately with H and E stain and 1% crystal violet stain. The stained sections were observed under microscope for identification and counting of MFs. Data obtained was statistically analyzed by using the Man-Whitney U test. A significant increase in number of MFs was observed in OED and OSCC in comparison with normal oral mucosa. There was a highly significant increase in number of MFs in crystal violet stained tissue sections when compared with H and E stain. Metaphase is the most commonly observed phase of mitosis in crystal violet stain when compared with H and E stain for all three groups. Crystal violet stain can be considered as selective stain for mitotic figures.

  7. Comparative assessment of seller's staining test (SST) and direct ...

    African Journals Online (AJOL)

    Background: Rabies causes 55, 000 annual human deaths globally and about 10,000 people are exposed annually in Nigeria. Diagnosis of animal rabies in most African countries has been by direct microscopic examination. In Nigeria, the Seller's stain test (SST) was employed until 2009. Before then, both SST and dFAT ...

  8. Image analysis of dye stained patterns in soils

    Science.gov (United States)

    Bogner, Christina; Trancón y Widemann, Baltasar; Lange, Holger

    2013-04-01

    Quality of surface water and groundwater is directly affected by flow processes in the unsaturated zone. In general, it is difficult to measure or model water flow. Indeed, parametrization of hydrological models is problematic and often no unique solution exists. To visualise flow patterns in soils directly dye tracer studies can be done. These experiments provide images of stained soil profiles and their evaluation demands knowledge in hydrology as well as in image analysis and statistics. First, these photographs are converted to binary images classifying the pixels in dye stained and non-stained ones. Then, some feature extraction is necessary to discern relevant hydrological information. In our study we propose to use several index functions to extract different (ideally complementary) features. We associate each image row with a feature vector (i.e. a certain number of image function values) and use these features to cluster the image rows to identify similar image areas. Because images of stained profiles might have different reasonable clusterings, we calculate multiple consensus clusterings. An expert can explore these different solutions and base his/her interpretation of predominant flow mechanisms on quantitative (objective) criteria. The complete workflow from reading-in binary images to final clusterings has been implemented in the free R system, a language and environment for statistical computing. The calculation of image indices is part of our own package Indigo, manipulation of binary images, clustering and visualization of results are done using either build-in facilities in R, additional R packages or the LATEX system.

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

    African Journals Online (AJOL)

    Rukevwe S. Abraka

    2016-09-14

    Sep 14, 2016 ... the tissue physical structure (for example, tightly versus loosely packed), and the amino acid composition of the elements of the tissue (Kiernan, 2002). ... colleagues (Sweat et al., 1964) to seek a better method. Picrosirius red F3BA was found to consistently stain thin collagen fibers, did not fade, and was ...

  10. The Stained Glass Paintings of Nigeria's Prime Artists, YCA Grillo

    African Journals Online (AJOL)

    Nneka Umera-Okeke

    Abstract. Many lamps same Light' investigates the place of agency in the transmutation of indigenous imageries in the art works of the pictorial turn. Through an investigation that entailed an empirical analysis of the works of two Nigerian prime stained glass artists, Yusuf Grillo and David Dale, this study established that in ...

  11. Activity staining method of chitinase on chitin agar plate through ...

    African Journals Online (AJOL)

    A method for detection of chitinase activity on chitin agar plate after polyacrylamide gel electrophoresis is described. Different staining dyes such as calcofluor white M2R, fluorescein isothiocyanate, rhodamine B, ruthenium red and congo red were separately incorporated in chitin agar plates. After running polyacrylamide ...

  12. Modelling multiple laser pulses for port wine stain treatment

    NARCIS (Netherlands)

    Verkruysse, W.; van Gemert, M. J.; Smithies, D. J.; Nelson, J. S.

    2000-01-01

    Many port wine stains (PWS) are still resistant to pulsed dye laser treatment. However, anecdotal information suggests that multiple-pulse laser irradiation improves patient outcome. Our aims in this note are to explain the underlying mechanism and estimate the possible thermal effects of multiple

  13. News from the Biological Stain Commission No. 5

    DEFF Research Database (Denmark)

    Lyon, H O; Dapson, R W

    2009-01-01

    In this fifth issue of News from the Biological Stain Commission (BSC), under the heading of Regulatory Affairs, the BSC's International Affairs Committee provides more information from the meeting of the International Standards Organization ISO/TC 212 Committee that took place on June 2-4, 2008 ...

  14. 'Many Lamps Same Light': The Stained Glass Paintings of Nigeria's ...

    African Journals Online (AJOL)

    Many lamps same Light' investigates the place of agency in the transmutation of indigenous imageries in the art works of the pictorial turn. Through an investigation that entailed an empirical analysis of the works of two Nigerian prime stained glass artists, Yusuf Grillo and David Dale, this study established that in spite of ...

  15. Amalgam stained dentin: a proper substrate for bonding resin composite?

    NARCIS (Netherlands)

    Scholtanus, J.D.

    2016-01-01

    Nowadays the use of dental amalgam is mostly abandoned and substituted by tooth colored resin composites that can be bonded to teeth tissues by adhesive techniques. The aim of this thesis was to find out whether dark stained dentin, as often observed after removal of amalgam restorations and

  16. Biocytin staining of glia and neurons in brain slices.

    Science.gov (United States)

    Kang, Jian

    2014-09-02

    This protocol describes the use of biocytin to visualize and distinguish the morphology of glia and neurons in rat brain slices. Patch pipettes are used to load biocytin into different cell types. The slices are subsequently fixed, stained, and mounted in preparation for imaging. © 2014 Cold Spring Harbor Laboratory Press.

  17. Credibility of Chromomycin A3 Staining in Prediction of Fertility

    Directory of Open Access Journals (Sweden)

    Roshanak Aboutorabi

    2009-01-01

    Full Text Available Background: Chromomycin A3 (CMA3 staining has been used to assess protamine deficiency.The aim of this study was to determine credibility of CMA3 along with semen parameters forassessment of fertility potential.Materials and Methods: Semen analysis and CMA3 staining were carried out on 234 fertile and178 subfertile individuals. Semen analysis was assessed according to WHO criteria. Protaminedeficiency was assessed by CMA3 staining.Results: Means, range of variables, coefficients of correlation and receiver operating characteristic(ROC analyses of semen parameters and protamine deficiency were determined. Mean values ofthree main sperm parameters and the percentage of sperm with negative CMA3 were significantlydifferent between fertile and sub fertile groups. The results of CMA3 assessment showed significantcorrelation with sperm density, percentage of motility and normal morphology in the total population,while in the subfertile group the results of CMA3 showed significant correlation with sperm densityand normal morphology. However in fertile men, the only significant correlation was observedbetween sperm with negative CMA3 and normal morphology. ROC analyses revealed that CMA3staining has a higher potential to predict fertility status, compared to semen parameters.Conclusion: Assessment of protamine deficiency could be considered as one of the complementarytests along with semen analysis for assessment of fertility.

  18. Fluorescent multiple staining and CASA system to assess boar ...

    African Journals Online (AJOL)

    The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA). The results obtained from this staining were compared with spermatozoa motility ...

  19. Black stain and dental caries in Filipino schoolchildren.

    NARCIS (Netherlands)

    Heinrich-Weltzien, R.; Monse, B.; Palenstein Helderman, W.H. van

    2009-01-01

    Black stain is defined as dark pigmented exogenous substance in lines or dots parallel to the gingival margin and firmly adherent to the enamel at the cervical third of the tooth crowns in the primary and permanent dentition. OBJECTIVES: This study was conducted to assess the prevalence of black

  20. Gross Morphology and Localization of Adenohypophyseal Cells in Camel (Camelus dromedarius Using A New Combination of Stains

    Directory of Open Access Journals (Sweden)

    S. A. S. Jaspal, Z. U. Rahman* and A. M. Cheema

    2011-01-01

    Full Text Available Thirty normal camels (Camelus dromedarius were selected for gross morphological and modified staining of anterior pituitary. Camels were divided in three age groups viz 2-4, 5-10 and above 10 years. Pituitary weight, length, width and circumference were recorded before preservation and at midsegittal cutting. Pituitary weight increased significantly as these animals grew older. Male had heavier pituitary as compared to female. Higher pituitary weight was observed in old as compared to young camel. Sections (4m of camel pituitary gland were stained with “Phosphotungstic acid haematoxylin-Orange G-Acid fuchsin-Light green” combination of dyes. This combination of acidic and basic dyes showed affinity to their respective adenohypophyseal cells and proved a suitable combination for differentiation of adenohypophyseal cells and architectural pattern of pituitary gland. Use of Lugol’s Iodine and sodium thiosulphate solution caused mercury fixation which ultimately enhanced the staining of camel adenohypophysis. The whole pituitary presented a brilliant appearance of clarity, enabling cell counts to be performed easily, purely with reference to the colors of adenohypophyseal cell types. This method can be applied for differential staining of adenohypophysis and with good cytology results to the hypophysis of many mammals. The method also provides a sharp contrast between cellular and connective tissue components. With this staining technique, the quantitative and qualitative characteristics of different adenohypophyseal cell types at various functional and hormonal stages, under certain physiological and pathological conditions can also be studied.

  1. Role of histochemical stains in differentiating hemangioma and vascular malformation

    Directory of Open Access Journals (Sweden)

    Ruchir Jitendra Patel

    2016-01-01

    Full Text Available Background: Benign vascular lesions such as vascular malformation and hemangioma at times pose difficulty in diagnosis both for clinicians and pathologists. Vascular malformations are difficult to treat while hemangiomas resolve spontaneously in most instances. There are instances when vascular malformations, especially arteriovenous malformations (AVMs have been misdiagnosed as hemangiomas and vice-versa. Clinical and radiological correlation with histopathological confirmation of these anomalies is important for the management of these lesionsAim: The aim was to study the histological characteristics of hemangiomas and vascular malformations and to study the utility of histochemical stains in their diagnosis. Materials and Methods: We retrospectively studied fifty cases retrieved from the records of Department of Pathology which were diagnosed as hemangioma (n=32 and vascular malformation (n=18 on Hematoxylin and Eosin (H and E stain over a period of 18 months. The cases were analyzed based on findings of histochemical stains such as Verhoeff-van Gieson (VVG, Masson's trichrome (MT, and toluidine blue. Results: After reviewing all the cases with the use of histochemical stains, two of the three cases originally diagnosed as hemangioma turned out to be AVM and one to be venous malformation. An increased number of intra-lesional nerves were found in 16 of 19 cases of AVM and in both cases of venous and lymphatic malformation. Hemangiomas did not show increase in nerve bundles. Mast cells were found to be increased in proliferating hemangiomas and pyogenic granulomas as compared to AVMs. Conclusion: Hemangiomas and vascular malformations should be clearly differentiated to reduce the risk of treatment failure and recurrence. With the use of histochemical stains such as VVG, MT and toluidine blue, the diagnostic difficulty can be reduced and definitive diagnosis is possible.

  2. A method for rapid quantitative assessment of biofilms with biomolecular staining and image analysis.

    Science.gov (United States)

    Larimer, Curtis; Winder, Eric; Jeters, Robert; Prowant, Matthew; Nettleship, Ian; Addleman, Raymond Shane; Bonheyo, George T

    2016-01-01

    The accumulation of bacteria in surface-attached biofilms can be detrimental to human health, dental hygiene, and many industrial processes. Natural biofilms are soft and often transparent, and they have heterogeneous biological composition and structure over micro- and macroscales. As a result, it is challenging to quantify the spatial distribution and overall intensity of biofilms. In this work, a new method was developed to enhance the visibility and quantification of bacterial biofilms. First, broad-spectrum biomolecular staining was used to enhance the visibility of the cells, nucleic acids, and proteins that make up biofilms. Then, an image analysis algorithm was developed to objectively and quantitatively measure biofilm accumulation from digital photographs and results were compared to independent measurements of cell density. This new method was used to quantify the growth intensity of Pseudomonas putida biofilms as they grew over time. This method is simple and fast, and can quantify biofilm growth over a large area with approximately the same precision as the more laborious cell counting method. Stained and processed images facilitate assessment of spatial heterogeneity of a biofilm across a surface. This new approach to biofilm analysis could be applied in studies of natural, industrial, and environmental biofilms.

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

    Science.gov (United States)

    Work, Thierry M.; Farah, Yael

    2014-01-01

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

  4. Cytological detection of spermatozoa: comparison of three staining methods.

    Science.gov (United States)

    Allery, J P; Telmon, N; Mieusset, R; Blanc, A; Rougé, D

    2001-03-01

    Sperm detection can be an important factor in confirming sexual assault in cases of rape. This paper compares three of the most commonly used staining methods cited in the scientific literature: Christmas tree. hematoxylin-eosin, and alkaline fuchsin. The population studied was composed of 174 consenting women seen at the Male Infertility Center in Toulouse. France. The date of their last sexual intercourse was accurately known. Alkaline fuchsin did not seem effective in detecting spermatozoa in vaginal samples. Compared with hematoxylin-eosin, Christmas tree stain appeared to be the most useful test in the first 72 h. Two external factors were associated with decreased detection of spermatozoa: time since in tercourse and sperm volume.

  5. 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...... results on the abundance, size distribution and bacterial colonization of ABSP therefore reflect general patterns of TEP. The abundance of ABSP in the size range 3-162 urn and retained by 3 um pore size filters averaged 3.6 ± 2.49 x 10s ml"1 (± SD), which is among the highest concentrations reported...... for comparable size spectra of TEP. On average, 35 % of ABSP (by number) were colonized by bacteria and 8.6 x 105 bacteria ml"1 lake water were attached to ABSP, which corresponds to 7% of the total bacterial abundance....

  6. Standard test method for determination of resistance to staining

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2004-01-01

    1.1 This test method is intended to determine the resistance to staining of ceramic tile surfaces. 1.2 The resistance to staining is determined by maintaining test solutions in contact with ceramic tile surfaces for a specified period of time. After exposure, the surface is cleaned in a defined manner, and the test specimens are inspected visually for change. 1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  7. [Standardization of Blastocystis hominis diagnosis using different staining techniques].

    Science.gov (United States)

    Eymael, Dayane; Schuh, Graziela Maria; Tavares, Rejane Giacomelli

    2010-01-01

    The present study was carried out from March to May 2008, with the aim of evaluating the effectiveness of different techniques for diagnosing Blastocystis hominis in a sample of the population attended at the Biomedicine Laboratory of Feevale University, Novo Hamburgo, Rio Grande do Sul. On hundred feces samples from children and adults were evaluated. After collection, the samples were subjected to the techniques of spontaneous sedimentation (HPJ), sedimentation in formalin-ether (Ritchie) and staining by means of Gram and May-Grünwald-Giemsa (MGG). The presence of Blastocystis hominis was observed in 40 samples, when staining techniques were used (MGG and Gram), while sedimentation techniques were less efficient (32 positive samples using the Ritchie technique and 20 positive samples using the HPJ technique). Our results demonstrate that HPJ was less efficient than the other methods, thus indicating the need to include laboratory techniques that enable parasite identification on a routine basis.

  8. Identification of active fluorescence stained bacteria by Raman spectroscopy

    Science.gov (United States)

    Krause, Mario; Beyer, Beatrice; Pietsch, Christian; Radt, Benno; Harz, Michaela; Rösch, Petra; Popp, Jürgen

    2008-04-01

    Microorganisms can be found everywhere e.g. in food both as useful ingredients or harmful contaminations causing food spoilage. Therefore, a fast and easy to handle analysis method is needed to detect bacteria in different kinds of samples like meat, juice or air to decide if the sample is contaminated by harmful microorganisms. Conventional identification methods in microbiology require always cultivation and therefore are time consuming. In this contribution we present an analysis approach to identify fluorescence stained bacteria on strain level by means of Raman spectroscopy. The stained bacteria are highlighted and can be localized easier against a complex sample environment e.g. in food. The use of Raman spectroscopy in combination with chemometrical methods allows the identification of single bacteria within minutes.

  9. USE OF VITAL STAINING IN STORED HUMAN PLATELETS MORPHOFUNCTIONAL ANALYSIS

    Directory of Open Access Journals (Sweden)

    M. S. Makarov

    2014-01-01

    Full Text Available Apheresis and pooled platelet concentrates, stored at 22°C during 5 days, were studied with morho-functional platelet rate analysis, based on vital cell staining and registration with fluorescent microscope. It was revealed that apheresis and pooled PC had, on the average, normal values of morphological and functional parameters. On the other hand, both PC kept MFPR of cells only for 2 days storage. Longer PC storage caused the significant decay of morphological and functional platelet parameters.

  10. Development of new staining technology "eastern blotting" using monoclonal antibody.

    Science.gov (United States)

    Morinaga, Osamu; Shoyama, Yukihiro

    2011-03-01

    Ginsenosides contained in Panax species were separated by silica gel TLC blotted to a polyvinylidene difluoride (PVDF) membrane which was dipped in a sodium periodide (NaIO(4)) solution and reacted with protein, preparing a ginsenoside-protein conjugate for binding a ginsenoside on a PVDF membrane. The blotted spots were stained by anti-ginsenoside-Rb1 monoclonal antibody (MAb) and anti-ginsenoside-Rg1MAb, respectively. The newly established immunostaining method, eastern blotting was applied for the determination of ginsenosides possessing protopanaxadiol and/or protopanaxatriol. Double staining of eastern blotting for ginsenosides using anti-ginsenoside-Rb1 MAb and anti-ginsenoside-Rg1 MAb promoted complete identification of ginsenosides in Panax species. This technique has been devised for the chromatographic separation and identification of ginsenosides using polyethersulfone (PES) membrane. It caused an acceptable separation of ginsenoside-Rb1, -Rc and -Rd in various ginseng extracts. Newly developed technique is quite simple and applies for immunoassay system. Ginsenosides separated using a PES membrane were directly treated with a NaIO(4) solution and then reacted with bovine serum albumin (BSA) for making a ginsenoside-protein conjugate. After the blocking, anti-ginsenoside-Rb1 MAb recognized a ginsenoside on a PES membrane and then a sec-ond antibody labeled with enzyme reacted to the first antibody. Finally a substrate was oxidized with the enzyme and de-veloped the staining of ginsenosides. The staining spots of ginsenosides on membrane were quantitatively evaluated by NIH Image indicating at least 62.5 ng of each ginsenoside-Rb1, -Rc and -Rd were detected with clarity. The determination range of three ginsenosides was from 0.125 to 2.0 µg of direct amount on PES membrane.

  11. Corneal blood staining following autologous blood injection for hypotony maculopathy.

    Science.gov (United States)

    Ayyala, R S; Urban, R C; Krishnamurthy, M S; Mendelblatt, D J

    1997-10-01

    Hypotony is a common complication following trabeculectomy in which antimetabolites are used. Autologous blood injection is an accepted form of treatment for hypotony that occurs secondary to overfiltration; however, injection into the filtering bleb has been associated with a rise in intraocular pressure for some patients with chronic postoperative hypotony. The authors describe a patient in whom corneal blood staining with raised intraocular pressure and loss of vision occurred as a result of autologous blood injection.

  12. Solid-Color Stains on Western Redcedar and Redwood Siding

    Science.gov (United States)

    Mark Knaebe

    2013-01-01

    You have decided to put wood siding on your new house. Several questions are probably going through your mind: “What’s the best type of wood?” “Should I use paint or stain?” “Should I apply the finish before or after I install the siding?”

  13. Cervicovaginal microbial flora in methenamine silver staining method

    Directory of Open Access Journals (Sweden)

    Noushin Afshar Moghaddam

    2006-12-01

    Full Text Available BACKGROUND: Vagina like all other mucosal organs owns its especial bacterial/microbial flora. Though may be pathogen in other circumstances, members of vaginal normal flora do not cause disease on healthy vaginal mucosa. In this study, we tried to determine the relationship between microscopic findings on Methenamine silver stained cervicovaginal smears and clinical symptoms. METHODS: A total of 389 cervicovaginal smears were examined cytologically from April to August 2005, among which 103 satisfactory smears of patients who were normally menstruating were subsequently selected. The originally Papanicolaou–stained smears were stained with Methenamine silver method. The cervicovaginal flora in symptomatic and asymptomatic patients was classified into four groups. The relationship between the type of genital flora and the presence of Candida or Actinomyces spp was also determined. Data were analyzed with SPSS software using Chi–square test. RESULTS: In 103 evaluated patients, 46 (44.7% were symptomatic and the rest were asymptomatic. The most prevalent genital microbial flora in both symptomatic (21.7% and asymptomatic (37.9% patients was type II (Lactobacilli. Microbial frequency differences were significant for types II (P = 0.034 and III (P = 0.039 in both groups. Coexistence of microbial flora of type I (P = 0.02 and type IV (P = 0.033 with Candida was statistically significant. Coexistence of all types of microbial flora with Actinomyces was not proved significant. CONCLUSIONS: Symptomatic women, except those with potential pathogens, tend to have Lactobacillus flora. Therefore, it is advisable that all Lactobacilli types be investigated through microbiological methods in symptomatic patients. In silver stained slides, there was a clear relationship between the type of vaginal microbial flora and the presence of Candida spp. KEY WORDS: Microbial flora, cervicovaginal smears, methenamine silver, symptomatic, asymptomatic.

  14. Adaptations of Goldner's Masson trichrome stain for the study of undecalcified plastic embedded bone.

    Science.gov (United States)

    Gruber, H E

    1992-01-01

    Specialized adaptations for application of Goldner's Masson trichrome stain to plastic embedded undecalcified bone specimens are presented. This stain can be used successfully on methyl-glycol methacrylate, glycol methacrylate and Spurr embedded bones. The stain affords the advantage of good cellular staining due to the hematoxylin component with concomitant sharp discrimination of mature bone matrix which stains green, immature new bone matrix which stains red, and calcified cartilage which stains very pale green. Use of red filters during photomicrography aids in bone-osteoid discrimination in black and white photographs.

  15. EVALUASI DALAM PEMBELAJARAN BAHASA ARAB DI STAIN PAMEKASAN

    Directory of Open Access Journals (Sweden)

    Maimun ---

    2011-07-01

    Full Text Available Evaluating study of arab Ianguage in STAIN Pamekasan represents one of the among study activity chain starts from study planning process, study execution and last is study evaluation. Evaluate at ability target (maharah language which must be evaluated, and also at elements (‘anashir language. As have been tolerated, that in arab Ianguage study at least there are four abilities (maharah which must be mastered by educative participant to get predicate that he is one who have ability in the arab Ianguage field. The Maharah is Maharah Istima'(ability to correct reading, maharah al-Kalam (ability to converse, maharah Kitabah (ability to write, and maharah al-Qiraah (ability to read. Methodologically, study evaluation process of arab Ianguage in STAIN Pamekasan researched by using the qualitative approach with research type of case which its target is all currator lecturers of Arab Ianguage subject. Research result is obtained as follows: a Step of assessment starting from preparation, execution, data-processing and follow-up, b evaluation form intended here is some instruments used by all lecturer to get information about college student efficacy, in STAIN Pamekasan the pattern mentioned becomes two, that is tes form and non tes form, c all lecturers more tend to using one approach of Ianguage tes, that is integrative approach

  16. Survival of Staphylococcus pseudintermedius in modified Romanowsky staining solutions.

    Science.gov (United States)

    Misan, Angus; Chan, Wei Yee; Trott, Darren; Hill, Peter B

    2017-08-01

    Stains that are used regularly for patient-side diagnosis to rapidly identify bacterial and fungal infections could become contaminated by common pathogens, such as Staphylococcus pseudintermedius, during slide immersion. To determine whether the inoculation of S. pseudintermedius into modified Romanowsky type stains (Quick Dip ® ) results in viable bacterial contamination and whether this is influenced by the addition of organic debris (canine hair and skin). A clinical isolate of S. pseudintermedius was inoculated into clean and organically contaminated Quick Dip ® solutions (methanol fixative, eosin, methylene blue), and positive (broth) and negative (bleach) controls. Each solution was tested for the presence of viable bacteria by counting the number of colony forming units (CFU/mL) at various time points. Solutions also were examined under high power microscopy to count the number of visible bacteria at each time point. Staphylococcus pseudintermedius was able to survive in the clean and contaminated Quick Dip ® stains for at least one hour, but by 24 h no viable bacteria remained. Survival of the bacteria was not supported in the fixative at any time point. Staphylococcus pseudintermedius remained visible under high power microscopy for up to 2 weeks in all organically contaminated solutions of the Quick Dip ® set. Staphylococcus pseudintermedius only remains viable in eosin and methylene blue for short periods of time, but the prolonged visibility of dead organisms could theoretically lead to the misdiagnosis of cytology samples. © 2017 ESVD and ACVD.

  17. Positive staining for cellulose in oral pulse granuloma.

    Science.gov (United States)

    Virkkunen, Sirke; Wolff, Henrik; Haglund, Caj; Højgaard, Casper; Winther, Jakob Rahr; Willemoës, Martin; Vogel, Ulla; Hagström, Jaana

    2017-04-01

    Oral pulse granuloma (OPG) is an oral inflammatory lesion characterized by the presence of hyaline rings with numerous multinucleated giant cells. The etiopathogenesis of this lesion is thus far unclear, as is the composition of the hyaline rings. Our aim was to investigate whether the hyaline rings contain cellulose. Using a newly developed staining method for cellulose, we studied 18 histologic samples diagnosed as OPG, in addition to 3 samples originally diagnosed as "normal" foreign body reactions. In our study, visualization of cellulose is based on its specific binding to the carbohydrate binding module of β-1,4-glycanase. All samples diagnosed as OPG were positive for cellulose staining localized in hyaline rings. In addition, 1 lesion (of 3), first diagnosed as a foreign body reaction without the presence of hyaline rings, was positive for cellulose by horseradish peroxidase staining. We show for the first time that cellulose is present in OPG lesions, indicating that cellulose might be the initial cause of formation of these lesions. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Meibomian orifices and Marx's line. Studied by triple vital staining.

    Science.gov (United States)

    Norn, M

    1985-12-01

    The ciliary margins of the lower lids have been vital stained by the lipid-specific Sudan III powder, fluorescein 0.1% and the bottom of the lacrimal river (Marx's line) by lissamine green 1% in 100 cases. The Meibomian orifices are situated in a straight row just in front of the Marx's line in the lipid phase. With increasing age (greater than 50 years) the orifices are more often displaced and also discharge their lipid in the depth of the aqueous phase. The number averaged 21.5 in the lipid phase and 1.7 in the aqueous phase. Active orifices staining with lipid were found in 45% of all orifices in normals, independent of age, and were increased in conjunctivitis in the lipid phase. Lissamine green-stained orifices were independent of age, phase and diagnosis. The anterior edge of Marx's line may run an irregular course in elderly normals (greater than 50 years), significantly more often in conjunctivitis and blepharitis.

  19. ``Gold corrosion'': red stains on a gold Austrian Ducat

    Science.gov (United States)

    Gusmano, G.; Montanari, R.; Kaciulis, S.; Montesperelli, G.; Denk, R.

    Stains of different colours have been observed on historic and modern gold coins in several countries. An Austrian Ducat at the Kunsthistorisches Museum in Vienna has developed some red spots on its surface over the years. The same defects have also been observed in modern coins of higher gold purity. The spots have been examined by OM, SEM, EDS, XPS and AES. Optical microscopy showed that ``red'' defects exhibit in fact a nuance of colours. The surface analysis put in evidence the presence in the stains, in addition to gold, of silver and sulphur. The values of the modified Auger parameter α' of silver correspond to those of Ag2S; thus, it can be assumed that the stains are composed of silver sulphide (Ag2S). It was not possible to determine whether the presence of silver on the surface is due to segregation towards the surface or to external particles of silver embedded in the matrix. Depth profiling performed on modern coins suffering from the same problem allowed us to demonstrate that the nuance of colours is due to the inhomogeneous thickness of the spots. Moreover, it was demonstrated that spots are formed by two layers: an outer layer of silver sulphide and an inner layer of silver.

  20. Stain guided mean-shift filtering in automatic detection of human tissue nuclei

    Directory of Open Access Journals (Sweden)

    Yu Zhou

    2013-01-01

    Full Text Available Background: As a critical technique in a digital pathology laboratory, automatic nuclear detection has been investigated for more than one decade. Conventional methods work on the raw images directly whose color/intensity homogeneity within tissue/cell areas are undermined due to artefacts such as uneven staining, making the subsequent binarization process prone to error. This paper concerns detecting cell nuclei automatically from digital pathology images by enhancing the color homogeneity as a pre-processing step. Methods: Unlike previous watershed based algorithms relying on post-processing of the watershed, we present a new method that incorporates the staining information of pathological slides in the analysis. This pre-processing step strengthens the color homogeneity within the nuclear areas as well as the background areas, while keeping the nuclear edges sharp. Proof of convergence for the proposed algorithm is also provided. After pre-processing, Otsu′s threshold is applied to binarize the image, which is further segmented via watershed. To keep a proper compromise between removing overlapping and avoiding over-segmentation, a naive Bayes classifier is designed to refine the splits suggested by the watershed segmentation. Results: The method is validated with 10 sets of 1000 × 1000 pathology images of lymphoma from one digital slide. The mean precision and recall rates are 87% and 91%, corresponding to a mean F-score equal to 89%. Standard deviations for these performance indicators are 5.1%, 1.6% and 3.2% respectively. Conclusion: The precision/recall performance obtained indicates that the proposed method outperforms several other alternatives. In particular, for nuclear detection, stain guided mean-shift (SGMS is more effective than the direct application of mean-shift in pre-processing. Our experiments also show that pre-processing the digital pathology images with SGMS gives better results than conventional watershed algorithms

  1. Analysis of the Microbiota of Black Stain in the Primary Dentition

    OpenAIRE

    Yue Li; Qian Zhang; Fangfei Zhang; Ruoxi Liu; He Liu; Feng Chen

    2015-01-01

    Black tooth stain is a characteristic extrinsic discoloration commonly seen on the cervical enamel following the contour of the gingiva. To investigate the relationship between black tooth stain and the oral microbiota, we used 16S rRNA gene sequencing to compare the microbial composition of dental plaque and saliva among caries-free children with and without black stain. Dental plaque and saliva, as well as black stain, were sampled from 10 children with and 15 children without black stain. ...

  2. A COMPARATIVE STUDY TO SEE THE UTILITY OF MODIFIED ULTRAFAST PAPANICOLAOU (MUFP STAIN OVER STANDARD PAP STAIN IN ROUTINE FNA SMEARS

    Directory of Open Access Journals (Sweden)

    Ruchi Khajuria

    2016-07-01

    Full Text Available BACKGROUND Pap stain is an excellent method to review the cytological specimen; however, it is time consuming and costly. Various modifications have been developed in Pap stain of which latest is Modified Ultrafast Pap (MUFP stain which is hybrid of the technique by Romanowsky and conventional Pap stain to reduce the staining time to 90 seconds. AIM Aim of this study was to assess the feasibility and applicability of MUFP stain in fine needle aspiration smears of various organs. MATERIAL AND METHODS This prospective study was carried out in the cytopathology laboratory of GMC, Jammu for a period of 6 months from December 2015 to May 2016. A total no of 200 specimens were collected. The samples included 80 lymph node aspiration samples, 40 thyroid FNA samples, 50 breast FNA samples, 25 soft tissue aspirations and 5 salivary gland aspirations. Two smears were kept for fixation in 95% ethanol for staining with standard Pap stain and 2 were air dried for MUFP staining. RESULTS A correct diagnosis was achieved in all the cases. Background was similar in both staining methods. However, well-preserved cell morphology, crisp nuclear outline, good overall staining were well seen with MUFP method when compared with the standard Pap method. CONCLUSION The findings of this study support the use of MUFP method in cytology laboratory over standard Pap method.

  3. Characterization of HIFU ablation using DNA fragmentation labeling as apoptosis stain

    Science.gov (United States)

    Anquez, Jeremie; Corréas, Jean-Michel; Pau, Bernard; Lacoste, François; Yon, Sylvain

    2012-11-01

    The goal of this work was to compare modalities to precisely quantify the extent of thermally induced lesions: gross pathology vs. histopathology vs. devascularization. Liver areas of 14 rabbits were targeted with HIFU and RF ablations in an acute study. Contrast enhanced computorized tomography (CE-CT) scan images were acquired two hours after HIFU and RF treatment to obtain the devascularized volumes of the livers. The animals were then euthanized and deep frozen. The livers were sliced and each slice was photographed and stacked yielding a volume of gross pathology. The volume VGP of the HIFU lesions were derived. The area AGP of the lesions were computed on a particular slice. The lesions were segmented as hypo intense (devascularized) regions on CE-CT images and their volumes VC were computed. The ratios VC/VGP were computed for all the HIFU lesions on all the 14 subjects with a mean value of 1.2. Histology was performed on the livers using Hematoxyline Eosine Staining (HES) and DNA Fragmentation labeling (TUNEL® technology) which characterizes apoptosis. Apoptotic regions of area AT were segmented on the images stained by TUNEL®. No necrosis was identified on the HES data. While TUNEL® did not mark the cores of the RF lesions as apoptotic, the periphery of HIFU and RF lesions was always recognized with TUNEL® as apoptotic. The ratio AGP/AT was computed. The mean value was 0.95 and 0.25 for HIFU and RF lesions respectively. These findings show that the devascularized territory seen on CE-CT scan coincide with the coagulated territories seen with gross pathology. Those actually correspond to cells in apoptosis. It is confirmed that HES stain does not show necrosis 2 hours after thermal ablation. TUNEL® technology for DNA fragmentation labeling appears as a useful marker for thermally induced acute lesions in the liver.

  4. Quantification of polysaccharides fixed to Gram stained slides using lactophenol cotton blue and digital image processing.

    Science.gov (United States)

    Ericksen, Bryan

    2015-01-01

    Dark blue rings and circles emerged when the non-specific polysaccharide stain lactophenol cotton blue was added to Gram stained slides. The dark blue staining is attributable to the presence of capsular polysaccharides and bacterial slime associated with clumps of Gram-negative bacteria.  Since all bacterial cells are glycosylated and concentrate polysaccharides from the media, the majority of cells stain light blue. The contrast between dark and light staining is sufficient to enable a digital image processing thresholding technique to be quantitative with little background noise. Prior to the addition of lactophenol cotton blue, the Gram-stained slides appeared unremarkable, lacking ubiquitous clumps or stained polysaccharides.  Adding lactophenol cotton blue to Gram stained slides is a quick and inexpensive way to screen cell cultures for bacterial slime, clumps and biofilms that are invisible using the Gram stain alone.

  5. Port wine stains: laser treatment and nursing management.

    Science.gov (United States)

    Walker, P S

    In 1960, an American physicist Theodore Maiman observed the first laser in action. Since then laser technology has progressed at an extraordinary rate. This article shows how one type of laser has touched the lives of thousands of people. The Candela tunable dye laser has given hope to people with port wine stain birthmarks. It also shows the duties and responsibilities of the nurse working with lasers and the possibility of the extended role of the nurse in the field of skin laser therapy.

  6. Weathering effects on materials from historical stained glass windows

    Directory of Open Access Journals (Sweden)

    García-Heras, M.

    2003-06-01

    Full Text Available A selection of materials (stained glasses, lead cames, support elements and putty from historical stained glass windows of different periods (13th-19th centuries have been studied. Optical microscopy, scanning electron microscopy, energy dispersive X-ray spectrometry and X-ray diffraction were used as characterization techniques. Degradation of historical stained glass windows is due to the particular chemical composition oftlie materials used for their production: stained glasses, lead network, metallic support elements and refilling putty. However, the presence of a given chemical composition is not the only factor involved in the degradation process. It is necessary the occurrence of other external factors that contribute to the development and progress of alteration problems in the materials mentioned above. The presence of gaseous pollution in the air produces a negative interaction with the surface of the stained glass windows materials. Firstly, the stained glasses and the grisailles begin a dealkalinisation process and a silica gel layer is formed during the early contact between the glasses and the wet environment. After that, insoluble salt deposits and corrosion crusts are formed as a consequence of a deeper chemical attack which results in a depolymerisation of the glass network. The lead cames and the metallic support elements are also altered by weathering. Such materials are oxidized and both pits and crusts appear on their surfaces. The transport of ions and other substances from the corrosion crusts of the metallic elements gives rise new deposits upon the stained glasses, which could intensify their own degradation processes. The putty experiments a noticeable shrinkage and cracking. Likewise, adverse environmental conditions favour the transport of putty substances towards the other materials of the stained glass window, thereby increasing the crusts thickness and adding elements that contribute to the total alteration of the

  7. Stain-free histopathology by programmable supercontinuum pulses

    DEFF Research Database (Denmark)

    Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry

    2016-01-01

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

  8. Visualization of latent blood stains using visible reflectance hyperspectral imaging and chemometrics.

    Science.gov (United States)

    Edelman, Gerda J; van Leeuwen, Ton G; Aalders, Maurice C

    2015-01-01

    The detection of latent traces is an important aspect of crime scene investigation. Blood stains on black backgrounds can be visualized using chemiluminescence, which is invasive and requires a darkened room, or near-infrared photography, for which investigators need to change filters manually to optimize contrast. We demonstrated the performance of visible reflectance hyperspectral imaging (400-720 nm) for this purpose. Several processing methods were evaluated: single wavelength bands, ratio images, principal component analysis (PCA), and "SIMPLe-to-use Interactive Self-modeling Mixture Analysis" (SIMPLISMA). Using these methods, we were able to enhance the contrast between blood stains and 12 different fabrics. On black cotton, blood dilutions were visible with a minimal concentration of 25% of whole blood. The hyperspectral camera system used in this study is portable and wireless, which makes it suitable for crime scene use. The described technique is noncontact and nondestructive, so all traces are preserved for further analysis. © 2014 American Academy of Forensic Sciences.

  9. Stain Resistance of Cotton Fabrics before and after Finishing with Admicellar Polymerization

    Directory of Open Access Journals (Sweden)

    Srinivas Hanumansetty

    2012-03-01

    Full Text Available Environmental concerns related to perfluoroctanoic acid (PFOA led to a re-examination of the methods for imparting stain resistance and stain repellency to textiles. Non-PFOA fluoropolymer finishes have been formed on cotton knits by admicellar polymerization, a surface analogue of emulsion polymerization. Fabric samples were characterized by a drop test, contact angle measurements, SEM, elemental analysis and durability studies. Stain resistance and stain release properties were assessed by reflectance and AATCC tests with results comparing favorably with swatches from commercially available garments. Admicellar polymerization enabled the formation of durable finishes that exhibited high performance in stain resistance and stain repellency.

  10. Authenticity screening of stained glass windows using optical spectroscopy

    Science.gov (United States)

    Meulebroeck, Wendy; Wouters, Hilde; Nys, Karin; Thienpont, Hugo

    2016-01-01

    Civilized societies should safeguard their heritage as it plays an important role in community building. Moreover, past technologies often inspire new technology. Authenticity is besides conservation and restoration a key aspect in preserving our past, for example in museums when exposing showpieces. The classification of being authentic relies on an interdisciplinary approach integrating art historical and archaeological research complemented with applied research. In recent decades analytical dating tools are based on determining the raw materials used. However, the traditional applied non-portable, chemical techniques are destructive and time-consuming. Since museums oftentimes only consent to research actions which are completely non-destructive, optical spectroscopy might offer a solution. As a case-study we apply this technique on two stained glass panels for which the 14th century dating is nowadays questioned. With this research we were able to identify how simultaneous mapping of spectral signatures measured with a low cost optical spectrum analyser unveils information regarding the production period. The significance of this research extends beyond the re-dating of these panels to the 19th century as it provides an instant tool enabling immediate answering authenticity questions during the conservation process of stained glass, thereby providing the necessary data for solving deontological questions about heritage preservation. PMID:27883056

  11. Dye staining and excavation of a lateral preferential flow network

    Directory of Open Access Journals (Sweden)

    A. E. Anderson

    2009-06-01

    Full Text Available Preferential flow paths have been found to be important for runoff generation, solute transport, and slope stability in many areas around the world. Although many studies have identified the particular characteristics of individual features and measured the runoff generation and solute transport within hillslopes, very few studies have determined how individual features are hydraulically connected at a hillslope scale. In this study, we used dye staining and excavation to determine the morphology and spatial pattern of a preferential flow network over a large scale (30 m. We explore the feasibility of extending small-scale dye staining techniques to the hillslope scale. We determine the lateral preferential flow paths that are active during the steady-state flow conditions and their interaction with the surrounding soil matrix. We also calculate the velocities of the flow through each cross-section of the hillslope and compare them to hillslope scale applied tracer measurements. Finally, we investigate the relationship between the contributing area and the characteristics of the preferential flow paths. The experiment revealed that larger contributing areas coincided with highly developed and hydraulically connected preferential flow paths that had flow with little interaction with the surrounding soil matrix. We found evidence of subsurface erosion and deposition of soil and organic material laterally and vertically within the soil. These results are important because they add to the understanding of the runoff generation, solute transport, and slope stability of preferential flow-dominated hillslopes.

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

    Science.gov (United States)

    Vilarigues, M.; Fernandes, P.; Alves, L. C.; da Silva, R. C.

    2009-06-01

    Stained glass fragments from the 15th, 16th and 20th centuries, belonging to Mosteiro de Santa Maria da Vitória, 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 (μ-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 μ-FTIR (Fourier Transform Infra Red) spectroscopy in combination with the data generated by μ-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.

  13. Authenticity screening of stained glass windows using optical spectroscopy

    Science.gov (United States)

    Meulebroeck, Wendy; Wouters, Hilde; Nys, Karin; Thienpont, Hugo

    2016-11-01

    Civilized societies should safeguard their heritage as it plays an important role in community building. Moreover, past technologies often inspire new technology. Authenticity is besides conservation and restoration a key aspect in preserving our past, for example in museums when exposing showpieces. The classification of being authentic relies on an interdisciplinary approach integrating art historical and archaeological research complemented with applied research. In recent decades analytical dating tools are based on determining the raw materials used. However, the traditional applied non-portable, chemical techniques are destructive and time-consuming. Since museums oftentimes only consent to research actions which are completely non-destructive, optical spectroscopy might offer a solution. As a case-study we apply this technique on two stained glass panels for which the 14th century dating is nowadays questioned. With this research we were able to identify how simultaneous mapping of spectral signatures measured with a low cost optical spectrum analyser unveils information regarding the production period. The significance of this research extends beyond the re-dating of these panels to the 19th century as it provides an instant tool enabling immediate answering authenticity questions during the conservation process of stained glass, thereby providing the necessary data for solving deontological questions about heritage preservation.

  14. Visualizing Peripheral Nerve Regeneration by Whole Mount Staining

    Science.gov (United States)

    Dun, Xin-peng; Parkinson, David B.

    2015-01-01

    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. PMID:25738874

  15. 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.

  16. Color stability of ceramic brackets immersed in potentially staining solutions

    Directory of Open Access Journals (Sweden)

    Bruna Coser Guignone

    2015-08-01

    Full Text Available OBJECTIVE: To assess the color stability of five types of ceramic brackets after immersion in potentially staining solutions.METHODS: Ninety brackets were divided into 5 groups (n = 18 according to brackets commercial brands and the solutions in which they were immersed (coffee, red wine, coke and artificial saliva. The brackets assessed were Transcend (3M/Unitek, Monrovia, CA, USA, Radiance (American Orthodontics, Sheboygan, WI, USA, Mystique (GAC International Inc., Bohemia, NY, USA and Luxi II (Rocky Mountain Orthodontics, Denver, CO, USA. Chromatic changes were analyzed with the aid of a reflectance spectrophotometer and by visual inspection at five specific time intervals. Assessment periods were as received from the manufacturer (T0, 24 hours (T1, 72 hours (T2, as well as 7 days (T3 and 14 days (T4 of immersion in the aforementioned solutions. Results were submitted to statistical analysis with ANOVA and Bonferroni correction, as well as to a multivariate profile analysis for independent and paired samples with significance level set at 5%.RESULTS: The duration of the immersion period influenced color alteration of all tested brackets, even though these changes could not always be visually observed. Different behaviors were observed for each immersion solution; however, brackets immersed in one solution progressed similarly despite minor variations.CONCLUSIONS: Staining became more intense over time and all brackets underwent color alterations when immersed in the aforementioned solutions.

  17. Antibody staining in C. elegans using "freeze-cracking".

    Science.gov (United States)

    Duerr, Janet S

    2013-10-14

    To stain C. elegans with antibodies, the relatively impermeable cuticle must be bypassed by chemical or mechanical methods. "Freeze-cracking" is one method used to physically pull the cuticle from nematodes by compressing nematodes between two adherent slides, freezing them, and pulling the slides apart. Freeze-cracking provides a simple and rapid way to gain access to the tissues without chemical treatment and can be used with a variety of fixatives. However, it leads to the loss of many of the specimens and the required compression mechanically distorts the sample. Practice is required to maximize recovery of samples with good morphology. Freeze-cracking can be optimized for specific fixation conditions, recovery of samples, or low non-specific staining, but not for all parameters at once. For antibodies that require very hard fixation conditions and tolerate the chemical treatments needed to chemically permeabilize the cuticle, treatment of intact nematodes in solution may be preferred. If the antibody requires a lighter fix or if the optimum fixation conditions are unknown, freeze-cracking provides a very useful way to rapidly assay the antibody and can yield specific subcellular and cellular localization information for the antigen of interest.

  18. Color stability and staining of silorane after prolonged chemical challenges

    DEFF Research Database (Denmark)

    de Jesus, Vivian CBR; Martinelli, Nata Luiz; Poli-Frederico, Regina Célia

    Objectives: The purpose of this study was to investigate the effect of prolonged chemical challenges on color stability and staining susceptibility of a silorane-based composite material when compared to methacrylate-based composites. Methods: Cylindrical specimens (n=24) were fabricated from...... methacrylate (Filtek Z250, 3M ESPE; Filtek Z350XT, 3M ESPE; Master Fill, Biodinâmica) or silorane-based (Filtek P90, 3M ESPE) composite materials. Initial color was registered in a spectrophotometer. Specimens were divided in four groups and individually stored at 37°C in 0.02N citric acid, 0.02N phosphoric...... acid, 75% ethanol or distilled water (control) for 7, 14, 21, and 180 days, when new measurements were performed. A staining test was performed (n=12) after 21 days of chemical challenge by immersion in coffee during 3 weeks at 37°C. Color changes (¿E) were characterized using the CIEL*a*b* color...

  19. Lugol staining and histological evaluation of esophageal mucosa in achalasia.

    Science.gov (United States)

    Yamamuro, Elisa Miki; Cecconello, Ivan; Iriya, Kiyoshi; El Ibrahim, Roberto; Rodrigues, Joaquim Gama; Pinotti, Henrique Walter

    2006-01-01

    Esophageal cancer in achalasia is often diagnosed in the advanced stage, which makes for a poor prognosis. Therefore, the aim of this study was to evaluate the macroscopic and histological features of the esophageal mucosa in order to improve the early detection of cancer. We studied the macroscopic features of esophageal mucosa using Lugol's solution and compared them with histological analysis of the entire mucosa in 20 esophagectomy specimens resected for achalasia. Intraepithelial neoplasia, when detected, was selected for DNA ploidy analysis through static cytometry. Macroscopically, the mucosa showed opacification and/or diffuse irregularities in 19 specimens. Advanced squamous cell carcinoma was diagnosed in 2 cases. Using Lugol, the esophageal mucosa acquired irregular brownish color. Clear unstained areas were circumscribed in 5 esophagi. They were macroscopically defined as ulcer, neoplasia (2 cases) and mucosal irregularities (2 cases). The histological analysis showed ulcer, squamous cell carcinoma (2 cases), Barrett's esophagus and esophagitis, respectively. The histological study of the stained mucosa revealed minute foci of DNA aneuploid intraepithelial neoplasia in 4 cases. Macroscopic examination using Lugol failed to identify minute foci of early carcinoma. The stained mucosa does not exclude the esophageal cancer risk in achalasia.

  20. Enhanced discrimination of benign from malignant prostatic disease by selective measurements of cleaved forms of urokinase receptor in serum

    DEFF Research Database (Denmark)

    Piironen, Timo; Haese, Alexander; Huland, Hartwig

    2006-01-01

    PAR(II-III) were higher in PCa than in benign disease. In men with tPSA between 2 and 10 microg/L, the combination of %fPSA with the ratio uPAR(I)/uPAR(I-III) had a greater area under the ROC curve (0.73) than did %fPSA (0.68). CONCLUSIONS: Specific measurements of different uPAR forms in serum improve...

  1. Circular Mixture Modeling of Color Distribution for Blind Stain Separation in Pathology Images.

    Science.gov (United States)

    Li, Xingyu; Plataniotis, Konstantinos N

    2017-01-01

    In digital pathology, to address color variation and histological component colocalization in pathology images, stain decomposition is usually performed preceding spectral normalization and tissue component segmentation. This paper examines the problem of stain decomposition, which is a naturally nonnegative matrix factorization (NMF) problem in algebra, and introduces a systematical and analytical solution consisting of a circular color analysis module and an NMF-based computation module. Unlike the paradigm of existing stain decomposition algorithms where stain proportions are computed from estimated stain spectra using a matrix inverse operation directly, the introduced solution estimates stain spectra and stain depths via probabilistic reasoning individually. Since the proposed method pays extra attentions to achromatic pixels in color analysis and stain co-occurrence in pixel clustering, it achieves consistent and reliable stain decomposition with minimum decomposition residue. Particularly, aware of the periodic and angular nature of hue, we propose the use of a circular von Mises mixture model to analyze the hue distribution, and provide a complete color-based pixel soft-clustering solution to address color mixing introduced by stain overlap. This innovation combined with saturation-weighted computation makes our study effective for weak stains and broad-spectrum stains. Extensive experimentation on multiple public pathology datasets suggests that our approach outperforms state-of-the-art blind stain separation methods in terms of decomposition effectiveness.

  2. A color-code for glycosaminoglycans identification by means of polyacrylamide gel electrophoresis stained with the cationic carbocyanine dye Stains-all.

    Science.gov (United States)

    Andrade, João Pedro Souza; Oliveira, Caroline Pacheco; Tovar, Ana Maria Freire; Mourão, Paulo Antonio de Souza; Vilanova, Eduardo

    2018-02-01

    Cationic dyes such as toluidin blue are commonly employed to visualize glycosaminoglycans (GAGs) on electrophoresis gels; however, the carbocyanine-based dye Stains-all have been increasingly used to stain the non-sulfated hyaluronic acid and other GAGs in submicrogram quantities. In this short communication, we demonstrate that Stains-all is able to stain the most common GAGs on polyacrylamide gels with distinct and contrasting colors in a reproducible manner. We also show that this staining method is useful to identify GAGs present both in mixtures and in submicrogram quantities. Therefore, Stains-all has shown to be useful in identifying GAGs on polyacrylamide gels with basis on their specific colors, at least on screening level. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Automatic and robust segmentation of endoscopic OCT images and optical staining.

    Science.gov (United States)

    Zhang, Jianlin; Yuan, Wu; Liang, Wenxuan; Yu, Shanyong; Liang, Yanmei; Xu, Zhiyong; Wei, Yuxing; Li, Xingde

    2017-05-01

    We report a generic method for automatic segmentation of endoscopic optical coherence tomography (OCT) images. In this method, OCT images are first processed with L1 -L0 norm minimization based de-noising and smoothing algorithms to increase the signal-to-noise ratio (SNR) and enhance the contrast between adjacent layers. The smoothed images are then formulated into cost graphs based on their vertical gradients. After that, tissue-layer segmentation is performed with the shortest path search algorithm. The efficacy and capability of this method are demonstrated by automatically and robustly identifying all five interested layers of guinea pig esophagus from in vivo endoscopic OCT images. Furthermore, thanks to the ultrahigh resolution, high SNR of endoscopic OCT images and the high segmentation accuracy, this method permits in vivo optical staining histology and facilitates quantitative analysis of tissue geometric properties, which can be very useful for studying tissue pathologies and potentially aiding clinical diagnosis in real time.

  4. 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.

  5. 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

    2017-06-21

    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.

  6. 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.

  7. 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

    Since the introduction of the methyl green-pyronin Y procedure as a differential histological stain more than 100 years ago, the method has become a histochemical procedure for differential demonstration of DNA and RNA. Numerous variants of the procedure have been suggested, and a number of hypot...... of methyl green and pyronin Y and by the pH of the staining solution....... 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...

  8. Out damn spot!--a study of the removal of dental material stains from cloth.

    Science.gov (United States)

    Cunningham, J L; Bolas, A

    1995-01-01

    Stains to white cotton caused by clinical dental materials were subjected to washing and dry cleaning processes. Most stains were successfully removed with the exception of a siloxane impression polymer.

  9. Flow microfluorometric and spectrophotofluorometric analysis of DNA staining in mammalian cells

    Energy Technology Data Exchange (ETDEWEB)

    Crissman, H.A.; Stevenson, A.P.; Kissane, R.J.

    1977-01-01

    The effects of various fixative agents, pH, ionic strength, stain concentration, and magnesium concentration on DNA staining with the antibiotics olivomycin, chromomycin A3, and mithramycin were examined with DNA in solution and in mammalian cells.

  10. A modified staining technique for arbuscular mycorrhiza compatible with molecular probes.

    Science.gov (United States)

    Pitet, M; Camprubí, A; Calvet, C; Estaún, V

    2009-02-01

    The effects of the different steps of the root staining on the arbuscular mycorrhizal (AM) fungal rDNA extraction and amplification have been assessed. The results obtained using molecular techniques are compared with those obtained from fresh, non-stained leek roots. A modified staining procedure that eliminates heating, the use of hydrochloric acid and trypan blue, has been proved to be the most adequate to observe the AM colonisation in different plant species with/without lignified roots allowing at the same time the subsequent rDNA extraction and amplification from the stained roots. The staining technique decreased the sensitivity of the process and a higher number of roots had to be used to obtain enough material for a positive amplification. The extraction and amplification process was reliable up to 3 days after staining. A week after staining, the amplification was not dependable and after 2 weeks there was no amplification from stained material.

  11. Staining ability and biocompatibility of brilliant blue G: preclinical study of brilliant blue G as an adjunct for capsular staining.

    Science.gov (United States)

    Hisatomi, Toshio; Enaida, Hiroshi; Matsumoto, Hiroyoshi; Kagimoto, Tadahisa; Ueno, Akifumi; Hata, Yasuaki; Kubota, Toshiaki; Goto, Yoshinobu; Ishibashi, Tatsuro

    2006-04-01

    To evaluate the effectiveness and biocompatibility of brilliant blue G (BBG) for capsular visualization for continuous curvilinear capsulorrhexis. The capsular staining ability of BBG was evaluated at graded concentrations of 10.0, 1.0, 0.5, 0.25, 0.1, and 0.01 mg/mL in enucleated pig's eyes. The biocompatibility of BBG was assessed in rat's eyes for 2 months. The eyes were analyzed using light, fluorescence, transmission electron, and scanning electron microscopy. TUNEL (terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling) was used to detect apoptotic cells, and endothelial cell counts were analyzed using scanning electron microscopy. The results were compared using indocyanine green and trypan blue. The BBG improved capsular visualization, and a complete capsulorrhexis could be performed. In the rat model, no apparent toxic effect was observed using biomicroscopy during 2 months. Histologically, BBG showed satisfactory biocompatibility. Apoptotic cell death of the endothelial cells was detected in only the trypan blue group. In contrast to BBG, indocyanine green and trypan blue showed degeneration of corneal endothelial cells using transmission and scanning electron microscopy. The BBG contributed to better capsular visualization and caused no apparent complications to the corneal endothelium.Clinical Relevance The BBG is effective and safe capsular staining for continuous curvilinear capsulorrhexis.

  12. Staining of non-carious human coronal dentin by caries dyes.

    Science.gov (United States)

    Boston, Daniel W; Liao, Janice

    2004-01-01

    This study tested the hypothesis that commercially available caries dyes stain non-carious human coronal dentin in freshly extracted teeth. Multiple sections were cut from 10 non-carious and two control carious teeth using a water-cooled saw. Each section was stained with one of five caries dyes. The location of staining, if any, was noted and the staining intensity was scored on a four-point scale. One of the sections from each tooth was subsequently decalcified and processed for observation under a light microscope using four histologic staining techniques to evaluate morphology, collagen distribution and bacterial content. The association between the stain intensity scores on the undecalcified sections and the five dyes was evaluated using the Kruskal-Wallis One-Way ANOVA by Ranks test. Outer carious dentin in the control specimens stained intensely with each of the five dyes. In the undecalcified, non-carious sections, all had at least one area of staining. However, this staining could be differentiated from the intensity of dye staining in the carious controls, except in two instances. The association between stain intensity scores and the five dyes was not statistically significant. In the histologic sections, numerous bacteria were seen within the dentinal tubules of carious lesions of the two control specimens; however, no bacteria were found in any of the sections from non-carious specimens. Histologically, no differences were observed in the morphology or staining pattern within mantle or circumpulpal dentin in areas stained with caries dye, and in only one unique instance within the main body of the dentin. These results suggest that the five dyes evaluated in this study can stain non-carious dentin, however, this stain can be differentiated from the staining of outer carious dentin in vitro.

  13. Maternal and fetal characteristics associated with meconium-stained amniotic fluid

    DEFF Research Database (Denmark)

    Balchin, Imelda; Whittaker, John C; Lamont, Ronald F

    2011-01-01

    To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF.......To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF....

  14. DEMONSTRATION OF GLOMERULAR LESIONS ON LIGHT MICROSCOPY WITH SILVER METHENAMINE AND MASSON'S TRICHROME STAINING

    OpenAIRE

    岡島, 馨; 山下, 隆司; 松永, 健司; 河原, 信吾; 中島, 充; 平, 康二; 上辻, 秀和

    1992-01-01

    In order to observe the precise morphological changes of glomeruli on lightmicroscopic level, silver methenamine and Masson's trichrome (SM-MT) staining was employed on renal biopsy sections from children with various glomerular diseases. With the SM-MT staining, basement membrane stains black, red blood cells red, freshfibrinoid material bright red and protein deposits are stained red in various shades depending on the composition and age of the deposits. There was a good contrast betweenbas...

  15. Staining clinical specimens for acid-fast bacilli by means of a mechanical conveyor system.

    Science.gov (United States)

    Heimer, G V; Joseph, N; Taylor, C E

    1978-02-01

    A Cyto-Tek staining machine designed for Papanicolau staining of cervical smears has been adapted for auramine staining of smears of sputum and other clinical specimens for acid-fast bacilli. The results compared favourably with those obtained by a manual method of staining. Contamination of negative smears by acid-fast bacilli from positive smears was not found. Since this study the machine has continued to be in routine use with a considerable saving in labour.

  16. Staining of rat thyroid parafollicular (C-) cells with the Sevier-Munger silver technique.

    Science.gov (United States)

    Wilander, E; Juntti-Berggren, L; Lundqvist, M; Grimelius, L

    1980-09-01

    The argyrophil silver stain of Grimelius and of Sevier-Munger was studied in rat thyroid parafollicular (C-) cells after formation and Bouin fixation. The strongest and most easily reproducible staining reaction was obtained in tissue fixed with Bouin's fluid and stained by the Sevier-Munger technique. The identify of the argyrophil cell as the calcitonin-containing C-cells was established by re-staining the same sections with calcitonin antibodies according to the method of Sternberger.

  17. Influence of stains on lesion contrast in the pits and fissures of tooth occlusal surfaces from 800-1600-nm

    Science.gov (United States)

    Almaz, Elias C.; Simon, Jacob C.; Fried, Daniel; Darling, Cynthia L.

    2016-02-01

    For over one hundred years, x-rays have served as a cornerstone of dentistry. Dental radiographic imaging technologies have constantly improved, however, detecting occlusal lesions remains as one of the greatest challenges due to the low sensitivity of radiographs and the overlap of enamel. Once detected, occlusal lesions have penetrated far into the dentin, necessitating invasive restorative treatment. The adoption of near-infrared (NIR) systems in dentistry introduces the potential for early detection of occlusal lesions. Commercially available NIR systems for intra-oral applications currently operate near 800-nm; however, extrinsic stains may interfere with the detection of demineralization of the underlying enamel surface. Higher wavelengths such as 1300-nm render stains nearly transparent and enhances the contrast of sound enamel to demineralized enamel. This novel finding promotes minimally invasive dentistry and allows oral health professionals the ability to detect, image, track, and monitor early lesions without repeated exposure to ionizing radiation nor invasive treatment.

  18. Identification of dentin phosphophoryn localization by histochemical stainings.

    Science.gov (United States)

    Takagi, Y; Fujisawa, R; Sasaki, S

    1986-01-01

    Phosphophoryn, the most abundant of the dentin non-collagenous proteins, has been considered to be related in function to the mineralization process. In the present study, identification of dentin phosphophoryn localization was attempted using newly developed, precautionary histological methods by which phosphophoryn was retained in the sections during the specimen preparation and stained selectively in situ. Phosphophoryn was found to be present widely in all of the calcified dentin except the mantle dentin, the external, first-formed portion of dentin, but was not found in the predentin, the inner, uncalcified layer of dentin. These results indicate that phosphophoryn is apparently related to the mineral phase of calcified dentin and that the mineralization process of mantle dentin, which is formed before the odontoblasts are fully differentiated, may be different from that of circumpulpal dentin.

  19. Conjugates of a Photoactivated Rhodamine with Biopolymers for Cell Staining

    Directory of Open Access Journals (Sweden)

    Sergei Yu. Zaitsev

    2014-01-01

    Full Text Available Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan (“Chitosan-PFD” and histone H1 (“Histone H1.3-PFD”. The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK. Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes (“caged” dyes for microscopic probing of biological objects. Thus, the synthesized “Chitosan-PFD” and “Histone H1-PFD” have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.

  20. Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections.

    Science.gov (United States)

    Brajkovic, Saska; Dupouy, Diego G; de Leval, Laurence; Gijs, Martin Am

    2017-08-01

    Frozen sections (FS) of tumor samples represent a cornerstone of pathological intraoperative consultation and have an important role in the microscopic analysis of specimens during surgery. So far, immunohistochemical (IHC) stainings on FS have been demonstrated for a few markers using manual methods. Microfluidic technologies have proven to bring substantial improvement in many fields of diagnostics, though only a few microfluidic devices have been designed to improve the performance of IHC assays. In this work, we show optimization of a complete pan-cytokeratin chromogenic immunostaining protocol on FS using a microfluidic tissue processor into a protocol taking <12 min. Our results showed specificity and low levels of background. The dimensions of the microfluidic prototype device are compatible with the space constraints of an intraoperative pathology laboratory. We therefore anticipate that the adoption of microfluidic technologies in the field of surgical pathology can significantly improve the way FSs influence surgical procedures.

  1. Surface discoloration of composite resins: Effects of staining and bleaching.

    Science.gov (United States)

    Poggio, Claudio; Beltrami, Riccardo; Scribante, Andrea; Colombo, Marco; Chiesa, Marco

    2012-09-01

    The purpose of this in vitro study was to evaluate surface discoloration of three microhybrid composite resins (Esthet•X HD, Clearfil AP-X, Gradia Direct) and five nanohybrid composite resins (Ceram•X, GC Kalore, G-aenial, Grandio, GrandioSO), after staining and bleaching procedures. The composite resins were polymerized with a curing light (Celalux II, Voco, Cuxhaven, Germany) into 160 silicon molds (6,4 mm in diameter and 2 mm in thickness) to obtain identical specimens. Twenty samples for each composite resin were prepared. The specimens were polished using an automated polishing machine with the sequence of 600-, 800-, 1000-grit abrasive paper under water irrigation. The specimens were immersed in tea and distilled water: the specimens were dipped for 20 min, once a day (every 24 h), for 14 days into the drinks. The specimens were then bleached with carbamide peroxide at 17% (Perfect Bleach-Voco). The color of specimens was measured with a spectrophotometer according to the CIE L(*)a(*)b(*) system after light-polymerization of composite resin specimens, after 7 days, after 14 days, and after bleaching. The color difference h index (DEab(*)) between each measurement was calculated. Statistical analysis was made using analysis of variance (ANOVA). All specimens showed a significant increase in staining with a similar trend and no significant differences between microhybrid and nanohybrid composite resins. After whitening procedures, materials tested showed both significant and unsignificant differences of the h index. Microhybrid and nanohybrid composite resins had similar in vitro surface discoloration in tea. After bleaching, discoloration was removed from some composite resins tested.

  2. Surface discoloration of composite resins: Effects of staining and bleaching

    Directory of Open Access Journals (Sweden)

    Claudio Poggio

    2012-01-01

    Full Text Available Background: The purpose of this in vitro study was to evaluate surface discoloration of three microhybrid composite resins (Esthet·X HD, Clearfil AP-X, Gradia Direct and five nanohybrid composite resins (Ceram·X, GC Kalore, G-aenial, Grandio, GrandioSO, after staining and bleaching procedures. Materials and Methods: The composite resins were polymerized with a curing light (Celalux II, Voco, Cuxhaven, Germany into 160 silicon molds (6,4 mm in diameter and 2 mm in thickness to obtain identical specimens. Twenty samples for each composite resin were prepared. The specimens were polished using an automated polishing machine with the sequence of 600-, 800-, 1000-grit abrasive paper under water irrigation. The specimens were immersed in tea and distilled water: the specimens were dipped for 20 min, once a day (every 24 h, for 14 days into the drinks. The specimens were then bleached with carbamide peroxide at 17% (Perfect Bleach-Voco. The color of specimens was measured with a spectrophotometer according to the CIE LFNx01aFNx01bFNx01 system after light-polymerization of composite resin specimens, after 7 days, after 14 days, and after bleaching. The color difference h index (DE abFNx01 between each measurement was calculated. Statistical analysis was made using analysis of variance (ANOVA. Results: All specimens showed a significant increase in staining with a similar trend and no significant differences between microhybrid and nanohybrid composite resins. After whitening procedures, materials tested showed both significant and unsignificant differences of the h index. Conclusions: Microhybrid and nanohybrid composite resins had similar in vitro surface discoloration in tea. After bleaching, discoloration was removed from some composite resins tested.

  3. Microfluidic Cell Cycle Analysis of Spread Cells by DAPI Staining

    Directory of Open Access Journals (Sweden)

    Jing Sun

    2017-01-01

    Full Text Available Single-cell cell cycle analysis is an emerging technique that requires detailed exploration of the image analysis process. In this study, we established a microfluidic single-cell cell cycle analysis method that can analyze cells in small numbers and in situ on a microfluidic chip. In addition, factors that influenced the analysis were carefully investigated. U87 or HeLa cells were seeded and attached to microfluidic channels before measurement. Cell nucleic DNA was imaged by 4′-6-diamidino-2-phenylindole (DAPI staining under a fluorescent microscope and subsequently fluorescent intensities of the cell nuclei DNA were converted to depict histograms for cell cycle phases. DAPI concentration, microscopic magnification, exposure time and cell number were examined for optimal cell cycle analysis conditions. The results showed that as few as a few hundred cells could be measured by DAPI staining in the range of 0.4–0.6 μg/mL to depict histograms with typical cell cycle phase distribution. Microscopic magnification during image acquisition, however, could distort the phase distribution. Exposure time did not significantly affect the cell cycle analysis. Furthermore, cell cycle inhibitor rapamycin treatment changed the cell cycle phase distribution as expected. In conclusion, a method for microfluidic single-cell cell cycle analysis of spread cells in situ was developed. Factors such as dye concentration and microscopic magnification had more influence on cell cycle phase distribution. Further studies will focus on detail differentiation of cell cycle phases and the application of such a method for biological meanings.

  4. Multi-stained whole slide image alignment in digital pathology

    Science.gov (United States)

    Déniz, Oscar; Toomey, David; Conway, Catherine; Bueno, Gloria

    2015-03-01

    In Digital Pathology, one of the most simple and yet most useful feature is the ability to view serial sections of tissue simultaneously on a computer monitor. This enables the pathologist to evaluate the histology and expression of multiple markers for a patient in a single review. However, the rate limiting step in this process is the time taken for the pathologist to open each individual image, align the sections within the viewer, with a maximum of four slides at a time, and then manually move around the section. In addition, due to tissue processing and pre-analytical steps, sections with different stains have non-linear variations between the two acquisitions, that is, they will stretch and change shape from section to section. To date, no solution has come close to a workable solution to automatically align the serial sections into one composite image. This research work address this problem to obtain an automated serial section alignment tool enabling the pathologists to simply scroll through the various sections in a single viewer. To this aim a multi-resolution intensity-based registration method using mutual information as a similarity metric, an optimizer based on an evolutionary process and a bilinear transformation has been used. To characterize the performance of the algorithm 40 cases x 5 different serial sections stained with hematoxiline-eosine (HE), estrogen receptor (ER), progesterone receptor (PR), Ki67 and human epidermal growth factor receptor 2 (Her2), have been considered. The qualitative results obtained are promising, with average computation time of 26.4s for up to 14660x5799 images running interpreted code.

  5. p53 immunohistochemical staining patterns in oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    G Dundy

    2016-09-01

    Full Text Available Background: Mutation of p53 gene is one of the most common events in oral carcinogenesis. Accumulation of p53 protein has also been detected in premalignant lesions.Materials and Methods:  This study included 40 biopsy samples, which were received in department of pathology, Sarojini Naidu Medical College, Agra, to ascertain p53 expression by immunohistochemically, in patients with oral squamous cell carcinomas and to correlate its expression with histological grade, different sites in oral cavity and tobacco intake/smoking habits.Results: Out of 40 biopsies of oral mucosa, 03 showed normal oral mucosa and 37 were diagnosed as squamous cell carcinoma (SCC, most patients were in 5th and 6th decade and majority (86.5% of oral SCC were males with buccal mucosa being the most common site. There was a statistically significant difference in p53 expression between oral SCC and normal oral mucosa (p value <0.05. Of total 37 cases, 12 cases were well differentiated type, 16 moderately differentiated and 09 of poorly differentiated type of SCC. In each category, about two thirds were positive for p53 staining. Out of total 37 cases of oral SCC, 64.9% were positive and 35.1% were negative for p53 expression, 34 cases had positive history of tobacco intake/smoking habits, of which 23 cases were positive while 11 cases were negative for p53 staining.Conclusion: Abnormal p53 protein was detected in 64.9% of oral squamous cell carcinoma, but not in normal oral mucosa. p53 expression was associated with malignant transformation of oral mucosa. 

  6. Combined beta-galactosidase and immunogold/silver staining for immunohistochemistry and DNA in situ hybridization

    NARCIS (Netherlands)

    van den Brink, W.; van der Loos, C.; Volkers, H.; Lauwen, R.; van den Berg, F.; Houthoff, H. J.; Das, P. K.

    1990-01-01

    A combination of beta-galactosidase enzyme and the immunogold/silver staining method was studied for evaluation of double-staining experiments. Applications are shown for immunohistochemical double staining using two monoclonal antibodies and for combined immunohistochemistry and DNA in situ

  7. A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria

    Science.gov (United States)

    Minnerath, Jeanne M.; Roland, Jenna M.; Rossi, Lucas C.; Weishalla, Steven R.; Wolf, Melissa M.

    2009-01-01

    Gram staining bacteria is a fundamental technique introduced in general biology and microbiology laboratory courses. Two common problems students encounter when Gram staining bacteria are (1) having a difficult time locating bacterial cells on the microscope slide and (2) over-decolorizing bacterial cells during the staining procedure such that…

  8. Complete staining of human spermatozoa and immature germ cells combined with phase contrast microscopy

    DEFF Research Database (Denmark)

    Michael, A Y; Drejer, J O; Bagger, P V

    1987-01-01

    A method combining Janus green B and Thymol blue stains the anterior part of the head, the nuclear membrane, middle piece, and tail of spermatozoa light green and the nucleus deep purple. The method provides excellent stained preparations for the evaluation of sperm morphology by phase contrast...... microscopy. It produces significantly less abnormal spermatozoa compared with the Papanicolaou stain....

  9. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty under...

  10. Detection of outdoor mould staining as biofinish on oil treated wood

    NARCIS (Netherlands)

    Nieuwenhuijzen, E.J. van; Sailer, M.F.; Gobakken, L.R.; Adan, O.C.G.; Punt, P.J.; Samson, R.A.

    2015-01-01

    Stains on wood are often unwanted in outdoor applications, dark stain formation however is essential to the development of a new protective, self-healing and decorative biotreatment for wood. The biotreatment is based on the formation of surface covering mould staining on linseed oil treated pine

  11. Comparison of quantitative light-induced fluorescence (QLF) and digital imaging applied for the detection and quantification of staining and stain removal on teeth.

    Science.gov (United States)

    Adeyemi, A A; Jarad, F D; Pender, N; Higham, S M

    2006-08-01

    This study compares the use of QLF with digital imaging in the detection and quantification of the development and removal of stain on teeth. Two experimental phases, tooth staining and tooth whitening, conducted in vitro on labial 12 mm(2) enamel windows made on ten extracted bovine teeth, developed stains in 6-min cycles (2 min in each solution) using artificial saliva, chlorhexidine and tea solutions and removed them using sodium perborate monohydrate in 2-min cycle monitored at the end of each cycle with QLF (Inspektor Research Systems, NL) and digital photography (Fuji, Japan). The stain values were quantified as DeltaQ derived from QLF and DeltaE from digital imaging. This was observed by the two methods correlated with Pearson correlation coefficient (r). Regression equations (R(2)) were also obtained. For both staining and stain removal there was a statistically significant (p<0.01) reverse correlation between DeltaQ values for QLF (r=-0.924, R(2)=85.4%) and DeltaE values for digital imaging (r=-0.994, R(2)=98.8%), respectively. QLF showed a high correlation with digital imaging as a technique for detecting and monitoring tooth stains and tooth whitening in vitro. The potential for QLF with further development as a tool for monitoring staining and whitening of teeth may be possible in vivo in addition to the diagnostic ability for caries detection.

  12. A novel contrast stain for the rapid diagnosis of pityriasis versicolor: A comparison of Chicago Sky Blue 6B stain, potassium hydroxide mount and culture

    Directory of Open Access Journals (Sweden)

    Nikita Lodha

    2015-01-01

    Full Text Available Background: The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. Aims and Objectives: This study was done to compare the utility of a novel contrast stain (CSB stain with KOH mount and culture. Materials and Methods: Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1 KOH mount and CSB stain for direct microscopic examination and (2 culture using Sabouraud′s dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen′s Kappa statistic was performed to determine consistency (agreement among the different modalities. Observations and Results: Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%, 92 (92% and 56 (56% patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%. Statistically significant fair agreement was found between CSB stain and KOH mount (94% with κ=0.38, P < 0.001. Negligible agreement was found between CSB stain and culture (66%, κ=0.199, P = 0.001 as well as between KOH mount and culture (64%, κ=0.051, P = 0.107. Conclusion: CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount.

  13. Aggrandizing oral submucous fibrosis grading using an adjunct special stain: A pilot study.

    Science.gov (United States)

    Reshma, V; Varsha, B K; Rakesh, P; Radhika, M B; Soumya, M; D'Mello, Sarah

    2016-01-01

    Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features though could be identified in routine hematoxylin and eosin (H and E) staining; they could be better appreciated in special stains. This pilot study is an attempt to identify a single special stain that can act as an adjunct to H and E stain to help grade this potentially malignant disease. To assess if special stains can improvise on differentiating the various histological changes seen in OSMF and to accordingly grade OSMF cases. Formalin-fixed paraffin-embedded tissue sections of OSMF-10 cases of each grade (n = 30). Three special stains: Van-Gieson, Mallory's trichrome and Masson trichrome. The results obtained were tabulated and statistically analyzed using Chi-square test. The thickness and degree of keratinization were best detected in Mallory's stain (100%) and were statistically significant; the subepithelial changes were better detected using special stains, especially Mallory's stain (100%). The changes in collagen fibers were better visualized in all three special stains but were not statistically significant. The changes in blood vessels were better detected in Van-Gieson's and Mallory's stain; the obtained results were statistically significant. The degree of fibrosis between muscle bundles could be detected in all the three special stains, but when compared the results were not statistically significant. The questionable areas of muscle degeneration, especially in deeper connective tissue were better detected in Mallory's (43%) and Masson's stain (43%) as compared to Van-Gieson stain (14%) and the results obtained were statistically significant. The inflammatory cells and dysplastic features are better visualized in routine H and E stains. Pathogenesis of OSMF is related to fibro-elastic and muscle degenerative changes in the connective tissue followed by secondary changes in

  14. A comparison of visible wavelength reflectance hyperspectral imaging and Acid Black 1 for the detection and identification of blood stained fingerprints.

    Science.gov (United States)

    Cadd, Samuel; Li, Bo; Beveridge, Peter; O Hare, William T; Campbell, Andrew; Islam, Meez

    2016-07-01

    Bloodstains are often encountered at scenes of violent crime and have significant forensic value for criminal investigations. Blood is one of the most commonly encountered types of biological evidence and is the most commonly observed fingerprint contaminant. Presumptive tests are used to test blood stain and blood stained fingerprints are targeted with chemical enhancement methods, such as acid stains, including Acid Black 1, Acid Violet 17 or Acid Yellow 7. Although these techniques successfully visualise ridge detail, they are destructive, do not confirm the presence of blood and can have a negative impact on DNA sampling. A novel application of visible wavelength hyperspectral imaging (HSI) is used for the non-contact, non-destructive detection and identification of blood stained fingerprints on white tiles both before and after wet chemical enhancement using Acid Black 1. The identification was obtained in a non-contact and non-destructive manner, based on the unique visible absorption spectrum of haemoglobin between 400 and 500nm. Results from the exploration of the selectivity of the setup to detect blood against ten other non-blood protein contaminants are also presented. A direct comparison of the effectiveness of HSI with chemical enhancement using Acid Black 1 on white tiles is also shown. Copyright © 2016 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.

  15. Coffee-stain growth dynamics on dry and wet surfaces

    Science.gov (United States)

    Boulogne, François; Ingremeau, François; Stone, Howard A.

    2017-02-01

    The drying of a drop containing particles often results in the accumulation of the particles at the contact line. In this work, we investigate the drying of an aqueous colloidal drop surrounded by a hydrogel that is also evaporating. We combine theoretical and experimental studies to understand how the surrounding vapor concentration affects the particle deposit during the constant radius evaporation mode. In addition to the common case of evaporation on an otherwise dry surface, we show that in a configuration where liquid is evaporating from a flat surface around the drop, the singularity of the evaporative flux at the contact line is suppressed and the drop evaporation is homogeneous. For both conditions, we derive the velocity field and we establish the temporal evolution of the number of particles accumulated at the contact line. We predict the growth dynamics of the stain and the drying timescales. Thus, dry and wet conditions are compared with experimental results and we highlight that only the dynamics is modified by the evaporation conditions, not the final accumulation at the contact line.

  16. Akreditasi Perpustakaan Perguruan Tinggi: Pengalaman Perpustakaan STAIN Kediri

    Directory of Open Access Journals (Sweden)

    Komarudin Komarudin

    2016-07-01

    Abstract; The importance of quality has been a concern of college library librarian. National Library has compiled standards can be used as a minimum level college library quality. A form of formal recognition of compliance with these standards is by accrediting library. Accreditation aims to improve accredited institution so useful to build a library quality. As stipulated by Law Decree(UU No. 43 of 2007 and Government Regulation (PPNo. 24 of 2014, the National Library has the National Library Accreditation Agency (LAP-N. Accredited certificate can obtain a library based on the number of components weighted values of service, cooperation, collection, organization of library materials, human resources, building / space and infrastructure, budget, library management and maintenance of library collection. The experience of STAIN Kediri library in carrying out the library accreditation including : make a plan of accreditation activities, form preparation team of accreditation, perform self assessments, set up support files, send a letter of application and data file support, assessment accreditation forms, prepare for site assessment and carry out the acreditation. The main thing is the accreditation is a culture of quality. Hope to obtain the best value of accreditation lies in the culture of quality.

  17. 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.

  18. Polarization-based non-staining cell detection.

    Science.gov (United States)

    Zhang, M; Ihida-Stansbury, K; Van der Spiegel, J; Engheta, N

    2012-11-05

    Polarization is an important characteristic of electromagnetic waves, which can not be detected by either the human visual system or traditional image sensors. Motivated by various animal species with polarization vision as well as by the prospect of improving the image quality of the imaging systems, we are exploring the potential of polarization for microscope imaging. The most powerful techniques for molecule monitoring requires complex preprocessing for labeling the sample with different dyes. In this paper, we propose a cell detection method using polarization imaging without any need for staining target cell samples with any chemical dye. The motivation for this work is to develop an optical imaging technique that is simple and that can be used on live cells. The polarization sensitivity of cell samples is studied in this paper. A definition for the quantity called "polarization deviation" is proposed in order to identify clearer the difference between target cells and the background. Based on the polarization deviation detection method, a three-parameter polarization imaging method is employed to further simplify the image capture procedure for the proposed label-free cell detection. A color imaging methodology based on the well-known color space is utilized in order to represent the captured polarization information using computer graphics.

  19. Preoperative iodine staining may complicate the demarcation of esophageal carcinoma.

    Science.gov (United States)

    Asada-Hirayama, Itsuko; Ono, Satoshi; Kodashima, Shinya; Niimi, Keiko; Mochizuki, Satoshi; Yamamichi, Nobutake; Fujishiro, Mitsuhiro; Matsusaka, Keisuke; Fukayama, Masashi; Koike, Kazuhiko

    2013-07-01

    A 53-year-old man was suspected of having an esophageal neoplasm. An endoscopic examination including Lugol chromoendoscopy suggested an esophageal squamous cell neoplasm limited to the lamina propria. A targeted biopsy showed atypical squamous cells, and an endoscopic submucosal dissection was performed 22 days after the previous endoscopy. Although a single 40 mm unstained area was observed by preoperative Lugol chromoendoscopy, intraoperative endoscopy revealed a 25 mm iodine-unstained area, with small unstained areas scattered on the oral side. We included the small unstained areas in the extent of the resection through assessment by preoperative endoscopy. Histopathologically, the tumor extent appeared to coincide with the preoperative assessment. Tumor cells were found in the basal-parabasal layers of the mucosa, in which small unstained areas were scattered, although the superficial layers exhibited well-differentiated cells containing glycogen in the cytoplasm. Although Lugol chromoendoscopy, which can induce chemical esophagitis, is widely used, re-epithelialization after mucosal damage by preoperative iodine staining may complicate the intraoperative demarcation of tumors.

  20. Ultrafast dynamics of epicocconone, a second generation fluorescent protein stain.

    Science.gov (United States)

    Chatterjee, Soumit; Burai, Tarak Nath; Karuso, Peter; Datta, Anindya

    2011-09-15

    Femtosecond upconversion experiment has been carried out for epicocconone and its butylamine adduct in acetonitrile and tert-butanol. An ultrafast component is found to dominate the decay of fluorescence of epicocconone in acetonitrile solution. Upon reacting with butylamine, a model for the epicocconone-protein adduct, this ultrafast component remains almost unaffected but an additional rise time occurs, indicating the formation of a highly emissive species from the locally excited state. This phenomenon is central to the extraordinary applications of epicocconone in biotechnology. The magnitude of the rise time of the butylamine adduct is similar to that of the longer component of the decay of epicocconone in acetonitrile, suggesting that the dynamics of epicocconone and its butylamine adduct are similar. The ultrafast component is slowed upon increasing the viscosity of the solvent. This results in a marked increase in quantum yield and suggests that it corresponds to rapid bond isomerization, leading to a nonradiative decay. Surprisingly, in water/sucrose mixtures, the ultrafast component remains unaffected but there is still an increase in quantum yield, suggesting that there are at least two nonradiative pathways, one involving bond isomerization and another involving proton transfer. The correct interpretation of these data will allow the design of second generation protein stains based on the epicocconone scaffold with increased quantum yields and photostability. © 2011 American Chemical Society

  1. Review of the extrinsic stain removal and enamel/dentine abrasion by a calcium carbonate and perlite containing whitening toothpaste.

    Science.gov (United States)

    Joiner, Andrew

    2006-08-01

    There has been an increase in the demand from consumers and patients for products that whiten teeth. To meet this demand, a whitening toothpaste containing calcium carbonate and perlite as the abrasive system and an efficacious fluoride source has recently been launched. The aim of the current paper is to review the toothpaste's stain removal efficacy and its effects on enamel and dentine wear. It has been shown to be effective at removing model extrinsic stain in vitro. Further, it has been shown to be more effective in removing naturally occurring extrinsic tooth stain than a silica non-whitening control toothpaste after two weeks of twice daily brushing in a parallel group, double-blind clinical study using 152 adult volunteers. In addition, the enhanced whitening effect did not give a clinically relevant level of wear to enamel or a significant increase in dentine wear compared to marketed non-whitening toothpaste formulations, as shown by using an in situ type model with ex vivo brushing.

  2. Blood culture gram stain, acridine orange stain and direct sensitivity-based antimicrobial therapy of bloodstream infection in patients with trauma

    Directory of Open Access Journals (Sweden)

    Behera B

    2010-01-01

    Full Text Available Purpose: The purpose of this study was to ascertain if the simple practice of Gram stain, acridine orange stain and direct sensitivity determination of positive blood culture bottles could be used to guide early and appropriate treatment in trauma patients with clinical suspicion of sepsis. The study also aimed to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. Findings from consecutive episodes of blood stream infection at an Apex Trauma centre over a 12-month period are summarized. Materials and Methods: A total of 509 consecutive positive blood cultures were subjected to Gram staining. AO staining was done in BacT/ALERT-positive Gram-stain negative blood cultures. Direct sensitivity was performed from 369 blood culture broths, showing single type of growth in Gram and acridine orange staining. Results of direct sensitivity were compared to conventional sensitivity for errors. Results: No ′very major′ discrepancy was found in this study. About 5.2 and 1.8% minor error rates were noted in gram-positive and gram-negative bacteria, respectively, while comparing the two methods. Most of the discrepancies in gram-negative bacteria were noted in β lactam - β lactamase inhibitor combinations. Direct sensitivity testing was not reliable for reporting of methicillin and vancomycin resistance in Staphylococci. Conclusions: Gram stain result together with direct sensitivity testing is required for optimizing initial antimicrobial therapy in trauma patients with clinical suspicion of sepsis. Gram staining and AO staining proved particularly helpful in the early detection of candidaemia.

  3. Hyperspectral imaging of the crime scene for detection and identification of blood stains

    Science.gov (United States)

    Edelman, G. J.; van Leeuwen, T. G.; Aalders, M. C. G.

    2013-05-01

    Blood stains are an important source of information in forensic investigations. Extraction of DNA may lead to the identification of victims or suspects, while the blood stain pattern may reveal useful information for the reconstruction of a crime. Consequently, techniques for the detection and identification of blood stains are ideally non-destructive in order not to hamper both DNA and the blood stain pattern analysis. Currently, forensic investigators mainly detect and identify blood stains using chemical or optical methods, which are often either destructive or subject to human interpretation. We demonstrated the feasibility of hyperspectral imaging of the crime scene to detect and identify blood stains remotely. Blood stains outside the human body comprise the main chromophores oxy-hemoglobin, methemoglobin and hemichrome. Consequently, the reflectance spectra of blood stains are influenced by the composite of the optical properties of the individual chromophores and the substrate. Using the coefficient of determination between a non-linear least squares multi-component fit and the measured spectra blood stains were successfully distinguished from other substances visually resembling blood (e.g. ketchup, red wine and lip stick) with a sensitivity of 100 % and a specificity of 85 %. The practical applicability of this technique was demonstrated at a mock crime scene, where blood stains were successfully identified automatically.

  4. Bulk acid-fast staining of sputum smears: time to end a taboo.

    Science.gov (United States)

    Kam, K M; Yip, C W; Tang, H S; Van Deun, A

    2009-09-01

    A high-throughput laboratory routinely performing fluorescence microscopy for acid-fast bacilli (AFB) smear with automated bulk staining. To determine the risk of false-positive AFB sputum smears from bulk staining showing as smear-positive, culture-negative specimens, or a decrease in smear- and culture-positives. Direct AFB smear and Löwenstein-Jensen culture were performed for a total of 39,350 routine sputum specimens. Of these, 6633 were randomly selected for individual AFB staining, while the remaining 32,717 were processed by bulk machine staining. Positives for smear and culture were compared. Overall, 111 specimens yielded a positive individually stained smear; of these, 100 (90.1%, 95%CI 83.0-95.0) were also culture-positive compared to 504/543 smear-positives after bulk staining (92.8%, 95%CI 90.6-95.0). The proportions of smear-positive, culture-negative and smear- and culture-positive specimens were respectively 1.8% vs. 2.2% and 90.1% vs. 92.8%, for individual and bulk staining (non-significant). The risk of transferring AFB from positive to negative smears during bulk AFB staining is negligible, if it occurs at all. Bulk staining should not be discouraged, as even in low-income countries this method will save significant resources, particularly manpower, and improve staining results in laboratories with a high workload.

  5. Discrimination of p53 immunohistochemistry-positive tumors by its staining pattern in gastric cancer

    Science.gov (United States)

    Ando, Koji; Oki, Eiji; Saeki, Hiroshi; Yan, Zhao; Tsuda, Yasuo; Hidaka, Gen; Kasagi, Yuta; Otsu, Hajime; Kawano, Hiroyuki; Kitao, Hiroyuki; Morita, Masaru; Maehara, Yoshihiko

    2015-01-01

    Immunohistochemistry staining of p53 is a cheap and simple method to detect aberrant function of p53. However, there are some discrepancies between the result of immunohistochemistry staining and mutation analysis. This study attempted to find a new definition of p53 staining by its staining pattern. Immunohistochemistry staining of p53 and TP53 gene mutation analysis were performed in 148 gastric cancer patients. Also SNP-CGH array analysis was conducted to four cases. Positive staining of p53 was observed in 88 (59.5%) tumors. Tumors with positive p53 staining showed malignant features compared to negative tumors. Mutation of TP53 gene was observed in 29 (19.6%) tumors with higher age and differentiated type. In positive p53 tumors, two types could be distinguished; aberrant type and scattered type. With comparison to TP53 gene mutation analysis, all the scattered type had wild-type TP53 gene (P = 0.0003). SNP-CGH array showed that scattered-type tumors had no change in the structure of chromosome 17. P53-scattered-type staining tumors may reflect a functionally active nonmutated TP53 gene. In interpretation of p53 immunohistochemistry staining, distinguishing p53-positive tumors by their staining pattern may be important in gastric cancer. PMID:25354498

  6. Staining of in vivo subsurface degradation in dental composites with silver nitrate

    Energy Technology Data Exchange (ETDEWEB)

    Mair, L.H. (Univ. of Liverpool (England))

    1991-03-01

    A previously reported technique for staining areas of degradation in dental composite restorations was evaluated in 51 removed restorations. The staining reagent was silver nitrate, which penetrated the degraded subsurface as ionic silver and was subsequently developed into colored deposits of metallic silver. Several artefacts were recognized that resulted in an apparent image of subsurface stain. Most importantly, the presence of a layer of adsorbed silver on the edge of the specimen exaggerated the extent of staining. In order for the true depth of stain to be determined, thin sections of the materials should first be examined with a stereomicroscope to distinguish any contribution from adsorbed silver on the specimen edge. With this regimen, no stain was present in 41% of the restorations, and in a further 30%, the depth of stain was less than 50 microns. In two composites, the depth of stain was greater than 900 microns, and in a number of specimens, localized stain was found in association with attrition scars. Energy-dispersive x-ray analysis indicated that the amount of silver present in the degraded layers was very small. Overall, the results indicated that the staining technique is useful in the study of composite degradation.

  7. Rapid Reticulin Fiber Staining Method is Helpful for the Diagnosis of Pituitary Adenoma in Frozen Section.

    Science.gov (United States)

    Noh, Songmi; Kim, Sun Ho; Cho, Nam Hoon; Kim, Se Hoon

    2015-05-01

    Approximately 90% of neoplasms found in the sellar region are adenoma of the pituitary gland. The use of frozen sections for the diagnosis of pituitary adenomas has an accuracy of 90% and is useful in evaluating complete tumor removal. However, it is sometimes difficult to diagnose pituitary adenomas using frozen sections because of the small sample size and marked artifact, and the contiguity of the pituitary adenoma with normal pituitary gland tissue. In this study, we evaluated the use of our modified reticulin stain to make correct decision in frozen section with reduced stain time and investigated the objective diagnostic criteria of pituitary adenoma with reticulin stain. We used Gomori's silver impregnation methods to stain reticulin fibers in frozen pituitary gland sections of 36 samples from 24 patients. We modified the conventional staining method by reducing the overall staining time. We diagnosed pituitary lesion according to our interpretation criteria and compared the results to those of the conventional method and findings of hematoxylin and eosin-stained slides. Reticulin fiber staining of normal adenohypophysis outlines the supporting stroma around the blood vessels and shows regular of the gland meshwork interconnecting the capillaries. In contrast, reticulin fiber staining of the adenomatous tissue shows loss of meshwork or frequent fragmentation. Our modified reticulin stain is more rapid than the established method and shows similar levels of accuracy. Independent evaluation by two pathologists showed discrepancies in diagnosis in four out of 36 cases with modified reticulin stain. Our rapid modified reticulin staining method for frozen sections may be useful as a diagnostic tool for pituitary adenomas and can complement routine hematoxylin and eosin staining.

  8. Modified fields' stain: ideal to differentiate Dientamoeba fragilis and Blastocystis sp.

    Science.gov (United States)

    Ragavan, Anitamalar Devi; Govind, Suresh Kumar

    2015-03-01

    Dientamoeba fragilis, a trichomonad parasite is usually found in the gastrointestinal tract of human, and it is known to be the cause for gastrointestinal disease. The parasite is globally distributed and mostly found in rural and urban areas. The parasite is found in humans and nonhuman primates such as the macaques, baboons, and gorillas. Often, the parasite is confused with another largely found organism in stools called Blastocystis sp. especially when seen directly under light microscopy on culture samples containing both parasites. Both sometimes are seen with two nuclei with sizes tending to be similar which complicates identification. Stools were collected fresh from nine previously diagnosed persons infected with D. fragilis who also were found to be positive for Blastocystis sp. Samples were then cultured in Loeffler's medium and were stained with Giemsa, iron hematoxylin, and modified Fields' (MF) stain, respectively. D. fragilis was differentiated from Blastocystis sp. when stained with MF stain by the presence of a thinner outer membrane with clearly demarcated nuclei in the center of the cell whilst Blastocystis sp. had a darker and thicker stained outer membrane with the presence of two nuclei. The staining contrast was more evident with modified Fields' stain when compared with the other two. The simplicity in preparing the stain as well as the speed of the staining procedure make MF stain an ideal alternate. The modified Fields' stain is faster and easier to prepare when compared to the other two stains. MF stain provides a better contrast differentiating the two organisms and therefore provides a more reliable diagnostic method to precisely identify one from the other especially when cultures show mixed infections.

  9. Tumor Budding Detection by Immunohistochemical Staining is Not Superior to Hematoxylin and Eosin Staining for Predicting Lymph Node Metastasis in pT1 Colorectal Cancer.

    Science.gov (United States)

    Okamura, Takuma; Shimada, Yoshifumi; Nogami, Hitoshi; Kameyama, Hitoshi; Kobayashi, Takashi; Kosugi, Shin-ichi; Wakai, Toshifumi; Ajioka, Yoichi

    2016-05-01

    Tumor budding is recognized as an important risk factor for lymph node metastasis in pT1 colorectal cancer. Immunohistochemical staining for cytokeratin has the potential to improve the objective diagnosis of tumor budding over detection based on hematoxylin and eosin staining. However, it remains unclear whether tumor budding detected by immunohistochemical staining is a significant predictor of lymph node metastasis in pT1 colorectal cancer. The purpose of this study was to clarify the clinical significance of tumor budding detected by immunohistochemical staining in comparison with that detected by hematoxylin and eosin staining. This was a retrospective study. The study was conducted at Niigata University Medical & Dental Hospital. We enrolled 265 patients with pT1 colorectal cancer who underwent surgery with lymph node dissection. Tumor budding was evaluated by both hematoxylin and eosin and immunohistochemical staining with the use of CAM5.2 antibody. Receiver operating characteristic curve analyses were conducted to determine the optimal cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining. Univariate and multivariate analyses were performed to identify the significant factors for predicting lymph node metastasis. Receiver operating characteristic curve analyses revealed that the cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining for predicting lymph node metastases were 5 and 8. On multivariate analysis, histopathological differentiation (OR, 6.21; 95% CI, 1.16-33.33; p = 0.03) and tumor budding detected by hematoxylin and eosin staining (OR, 4.91; 95% CI, 1.64-14.66; p = 0.004) were significant predictors for lymph node metastasis; however, tumor budding detected by CAM5.2 staining was not a significant predictor. This study was limited by potential selection bias because surgically resected specimens were collected instead of endoscopically resected specimens. Tumor budding detected by

  10. Ziehl-Neelsen staining technique can diagnose paragonimiasis.

    Science.gov (United States)

    Slesak, Günther; Inthalad, Saythong; Basy, Phadsana; Keomanivong, Dalaphone; Phoutsavath, Ounheaun; Khampoui, Somchaivang; Grosrenaud, Aude; Amstutz, Vincent; Barennes, Hubert; Buisson, Yves; Odermatt, Peter

    2011-05-01

    We evaluated the Ziehl-Neelsen staining (ZNS) technique for the diagnosis of paragonimiasis in Laos and compared different modifications of the ZNS techniques. WE APPLIED THE FOLLOWING APPROACH: We (1) examined a paragonimiasis index case's sputum with wet film direct examination (WF) and ZNS; (2) re-examined stored ZNS slides from two provinces; (3) compared prospectively WF, ZNS, and formalin-ether concentration technique (FECT) for sputum examination of patients with chronic cough; and (4) compared different ZNS procedures. Finally, we assessed excess direct costs associated with the use of different diagnostic techniques. Paragonimus eggs were clearly visible in WF and ZNS sputum samples of the index case. They appeared brownish-reddish in ZNS and were detected in 6 of 263 archived ZNS slides corresponding to 5 patients. One hundred sputum samples from 43 patients were examined with three techniques, which revealed that 6 patients had paragonimiasis (13 positive samples). Sensitivity per slide of the FECT, ZNS and the WF technique was 84.6 (p = 0.48), 76.9 (p = 0.25) and 61.5% (p = 0.07), respectively. Percentage of fragmented eggs was below 19% and did not differ between techniques (p = 0.13). Additional operational costs per slide were 0 (ZNS), 0.10 US$ (WF), and 0.79 US$ (FECT). ZNS heated for five minutes contained less eggs than briefly heated slides (29 eggs per slide [eps] vs. 42 eps, p = 0.01). Bloodstained sputum portions contained more eggs than unstained parts (3.3 eps vs. 0.7 eps, p = 0.016). Paragonimus eggs can easily be detected in today's widely used ZNS of sputum slides. The ZNS technique appears superior to the standard WF sputum examination for paragonimiasis and eliminates the risk of tuberculosis transmission. Our findings suggest that ZNS sputum slides should also be examined routinely for Paragonimus eggs. ZNS technique has potential in epidemiological research on paragonimiasis.

  11. AUTHENTIC MATERIALS IN EXTENSIVE READING CLASS AT STAIN PONOROGO

    Directory of Open Access Journals (Sweden)

    Dhinuk Puspita Kirana

    2013-12-01

    Full Text Available It is widely believed that English Foreign Language (EFL learners need to develop their language proficiency by getting so much input. Moreover, students need to be familiarized with the real English us­age where real forms of communication and cultural knowledge are crucially exposed. Teaching through authentic materials will make the learners feel that they are learning a real language which is used by the real native speakers for real communication. incorporating au­thentic materials helps students acquire an effective communicative competence in the language focus. The research intended to describe the implementation of authentic materials in extensive reading class, the problems arise and the students’ responses toward the authen­tic materials in extensive reading class. The design of the research was Descriptive Qualitative method and the research subject was the lecturer of Extensive Reading class and 33 students in B class of the fourth semester of STAIN Ponorogo who took Extensive Read­ing subject. The instruments used were in the form of observation sheet, interview guideline and questionnaire. The implementation of authentic materials in extensive reading class covered some procedures into three main phases namely (1 Pre­ Activity, (2 Main­ Activity and (3 Post­Activity. The activities in main activity are as follows: (a Pre­ Activity; (b Whilst ­Activity; and (3 The language focus stage. There were problems arose during the implementation in terms of complicated planning, more time allocation and some disinterested students. Finally, the students showed significantly positive attitude toward the implementation of authentic materials in extensive reading class.

  12. The Stained Glass Paintings of Nigeria's Prime Artists, YCA Grillo

    African Journals Online (AJOL)

    Nneka Umera-Okeke

    That environment is enhanced by the visual culture implicit in story-telling traditions; and her world of art, crafts, body and architectural decoration traditions. However, Dale in a most profound way deepens his iconographical detours by a universalisation of imagery. Dale goes beyond the Nigerian art world, basking thereof ...

  13. The chemical stain removal properties of 'whitening' toothpaste products: studies in vitro.

    Science.gov (United States)

    Sharif, N; MacDonald, E; Hughes, J; Newcombe, R G; Addy, M

    2000-06-10

    A considerable number of toothpastes are available as tooth whitening products. Most appear to contain ingredients that might remove extrinsic stains rather than change natural tooth colour. Extrinsic stain removal could be achieved by physical or chemical means. The purpose of this study was to measure the chemical stain removal properties of a range of whitening toothpaste products and experimental formulations using a standardised method in vitro. 5 separate studies were conducted involving a total of 39 agents of which 28 were whitening products, 7 were experimental formulations, 2 were oxidising mouthrinses used as positive controls, 1 was a popular fluoride toothpaste product as a benchmark control, and 1 was water as the negative control. The formulations and controls varied in each study. The stain model was saliva/chlorhexidine/tea stain developed on optically clear acrylic to an optical density of at least 2.0. Groups of stained specimens were exposed to standard slurries or solutions of each test agent for 1 minute periods up to 5 minutes. Optical density readings were taken at each 1 minute time point. Analyses were based on per cent stain remaining after 5 minutes and time to 75% stain remaining. 3 toothpaste products achieved 100% stain removal by 5 minutes; 2 of these in 3 out of 4 studies in which they were used. 4 experimental formulations also achieved 100% stain removal. In general agents with high total stain removal also had short times to 75% stain remaining. The majority of agents tested had low total chemical stain removal and prolonged times to 75% stain remaining. A few agents were little different from water and several similar in effect to the conventional fluoride toothpaste. This method in vitro tests agents under the best case scenario conditions for chemical stain removal. Only a small number of the whitening toothpaste products have good chemical stain removal potential; the majority are unlikely to achieve their claimed benefits

  14. Standardization of stain used for diagnosing erythrocytic inclusion body syndrome (EIBS)

    Science.gov (United States)

    1987-01-01

    Erythrocytic inclusion body syndrome (EIBS), a viral erythrocytic necrosis (VEN)-like disease, has been observed in several areas in the Northwest. This virus disease is clinically diagnosed by microscopic examination of blood smears for intracytoplasmic erythrocytic inclusion bodies. Fish biologists involved in EIBS diagnostic work have been using several types of hematological stains. It became apparent that standardization of the staining procedure was needed. Comparative tests were conducted on blood smears and kidney imprints with the following commonly used blood stains: (1) Leishman-Giesma, (2) Pinacyanol chloride, (3) Powell 's Giemsa, (4) Harleco's Giemsa, (5) Diff Quik differential stain, (6) Wright's.Pinacyanol chloride stain was found to be the most consistent. The following staining procedure is recommended.

  15. Unsupervised color normalisation for H and E stained histopathology image analysis

    Science.gov (United States)

    Celis, Raúl; Romero, Eduardo

    2015-12-01

    In histology, each dye component attempts to specifically characterise different microscopic structures. In the case of the Hematoxylin-Eosin (H&E) stain, universally used for routine examination, quantitative analysis may often require the inspection of different morphological signatures related mainly to nuclei patterns, but also to stroma distribution. Nevertheless, computer systems for automatic diagnosis are often fraught by color variations ranging from the capturing device to the laboratory specific staining protocol and stains. This paper presents a novel colour normalisation method for H&E stained histopathology images. This method is based upon the opponent process theory and blindly estimates the best color basis for the Hematoxylin and Eosin stains without relying on prior knowledge. Stain Normalisation and Color Separation are transversal to any Framework of Histopathology Image Analysis.

  16. COMPARISON OF PERMANENT STAINING METHODS FOR THE LABORATORY DIAGNOSIS OF TRICHOMONIASIS.

    Science.gov (United States)

    Menezes, Camila Braz; Mello, Mariana dos Santos; Tasca, Tiana

    2016-01-01

    Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis.

  17. An improved silver staining procedure for schizodeme analysis in polyacrylamide gradient gels

    Directory of Open Access Journals (Sweden)

    Antonio M. Gonçalves

    1990-03-01

    Full Text Available A simple protocol is described for the silver staining of polyacrylamide gradient gels used for the separation of restriction fragments of kinetoplast DNA [schizodeme analysis of trypanosomatids (Morel et al., 1980]. The method overcomes the problems of non-uniform staining and strong background color which are frequently encountered when conventional protocols for silver staining of linear gels. The method described has proven to be of general applicability for DNA, RNA and protein separations in gradient gels.

  18. Staining and in vitro toxicity of dithizone with canine, porcine, and bovine islets.

    Science.gov (United States)

    Clark, S A; Borland, K M; Sherman, S D; Rusack, T C; Chick, W L

    1994-01-01

    Dithizone (DTZ) is a recognized diabetogenic agent in vivo, and a supravital stain commonly used for identification of islets to be used for transplantation. In the present studies, we compared DTZ staining of freshly isolated and cultured canine, bovine, and porcine islets, and the effect of DTZ on the function and viability of islets. Incubation with DTZ resulted in staining of canine and porcine islets, but no discernible staining with bovine islets. Insulin content of porcine, canine, and bovine islet was 2.0 +/- 0.2, 2.2 +/- 0.3, and 1.9 +/- 0.2 mU/EIN, indicating a lack of correspondence of DTZ staining and insulin content. Seven days of culture with canine islets resulted in > or = 50% reduction of DTZ stained cells. Exposure to DTZ at 50 micrograms/mL resulted in a maximal number of stained cells in preparations of purified islets (80-85%; counted after dispersion), a lower percentage of cells stained faintly at 20 micrograms/mL (50-55%), with no discernible staining at 10 micrograms/mL. Prolonged exposure of islets (4-48 h) to 20 micrograms/mL DTZ led to reduced insulin secretion and islet cell death. Incubation of canine or porcine islets with 100 micrograms/mL of DTZ for 0.5 h resulted in a dramatic loss of viability and diminished insulin secretory function, which was not reversed with continued culture. The concentration dependence of toxic effects paralleled the concentration dependence of cellular staining. The minimally effective staining concentration (20 micrograms/mL) also resulted in a loss of viability. An additional assessment of DTZ toxicity was made using the RIN-38 beta-cell line, which shows no discernible staining with DTZ.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. DYES AND CHEMICALS USED IN BIOMATERIAL STUDY AS STAINS FOR INVERTEBRATES

    OpenAIRE

    LONKAR, S; KEDAR, G; KOTANGALE, J; KALE, MANOJ

    2011-01-01

    Most of the dyes used in histology and cytology are manufactured for use in the textile industry, printing, food, cosmetics and other colorant industries. Chemicals used in the study include dyes and stains; the stains used are eosin, acetocarmine, rose bengal, magnesium chloride, magnesium sulphate, cocaine, menthol, propylene phenoxetol, osmic mercuric chloride, mercuric chloride, acetic acid, glycerin alcohol, phenoxetol, nitric acid, potassium cyanide. The dyes and stains as chemicals are...

  20. Niagara sky blue 6B--a new stain for granulocytic cells.

    Science.gov (United States)

    Kass, L

    1980-12-01

    Using an acidified solution of Niagara sky blue 6B, granules in mature granulocytic cells from peripheral blood and bone marrow stained bright blue to blue-green. Granules in immature granulocytes like myelocytes and promyelocytes stained purple. In leukemic myeloblasts and leukemic monocytes, granular-appearing structures stained purple. In leukemic lymphoblasts, purple granular structures were not visualized. As such, Niagara sky blue 6B can be used to identify granulocytic cells at all maturational stages.

  1. Nile blue A as a fluorescent stain for poly-beta-hydroxybutyrate.

    OpenAIRE

    Ostle, A G; Holt, J G

    1982-01-01

    Poly-beta-hydroxybutyrate granules exhibited a strong orange fluorescence when stained with Nile blue A. Heat-fixed cells were treated with 1% Nile blue A for 10 min and were observed at an excitation wavelength of 460 nm. Glycogen and polyphosphate did not stain. Nile blue A appears to be a more specific stain for poly-beta-hydroxybutyrate than Sudan black B.

  2. Detection of Entamoeba histolytica in experimentally induced amoebic liver abscess: comparison of three staining methods.

    Science.gov (United States)

    Ning, Tan Zi; Kin, Wong Weng; Mustafa, Shaymoli; Ahmed, Arefuddin; Noordin, Rahmah; Cheong, Tan Gim; Alfonso, Olivos-Garcia; Huat, Lim Boon

    2012-01-01

    To compare the efficacy of three different tissue stains, namely haematoxylin and eosin (H&E), periodic-acid Schiff (PAS) and immunohistochemical (IHC) stains for detection of Entamoeba histolytica (E. histolytica) trophozoites in abscessed liver tissues of hamster. Amoebic liver abscess was experimentally induced in a hamster by injecting 1 × 10(6) of axenically cultured virulent E. histolytica trophozoites (HM1-IMSS strain) into the portal vein. After a week post-inoculation, the hamster was sacrificed and the liver tissue sections were stained with H&E, PAS and IHC stains to detect the amoebic trophozoite. The three stains revealed tissue necrosis and amoebic trophozoites, but with varying clarity. H&E and PAS stained the trophozoites pink and magenta, respectively, however it was difficult to differentiate the stained trophozoites from the macrophages because of their similarity in size and morphology. On the other hand, IHC stain revealed distinct brown appearance of the trophozoites in the infected liver tissues. It can be concluded that out of the three stains, IHC is the best for identification of E. histolytica trophozoites in tissue sections.

  3. Skeletal examination by double staining for ossified bone and cartilaginous tissue.

    Science.gov (United States)

    Schneider, Steffen

    2013-01-01

    The assessment of developmental toxicology data is a critical aspect of hazard evaluation for pharmaceuticals and environmental chemicals. Skeletal examination is an essential part of prenatal developmental toxicity studies of chemicals as well as pesticides and comprises evaluation of both cartilaginous and ossified skeletal components. Various techniques are published in the literature to process and double-stain skeletons of common laboratory animals which are all based on staining of the cartilage with Alcian Blue and staining of ossified bones with Alizarin Red S along with maceration (clearing) of the surrounding soft tissue. The staining of the cartilage allows the examination to assess ossified structures and their underlying cartilage in a single step.

  4. A standard tissue as a control for histochemical and immunohistochemical staining

    Science.gov (United States)

    Otali, D; Fredenburgh, J; Oelschlager, DK; Grizzle, WE

    2017-01-01

    The variable quality of histochemical and immunohistochemical staining of tissues may be attributed to pre-analytical and analytical variables. Both categories of variables frequently are undefined or inadequately controlled during specimen collection and preparation. Pre-analytical variables may alter the molecular composition of tissues, which results in variable staining; such variations may cause problems when different tissues are used as staining controls. We developed a standard tissue for use as a staining control. Our standard tissue contains five components: 1) nine combined human cell lines mixed with stroma from human spleen; 2) a squamous cancer cell line, A431; 3) fungus; 4) transverse sections of the mosquitofish and 5) normal human spleen. The first three components were embedded in HistoGel™ and all components were processed to paraffin and used to construct a single standard paraffin block. The muscles of mosquitofish and arteries of the spleen are positive controls for eosin staining, while other tissues are useful for assessing hematoxylin staining. The mosquitofish tissues also are excellent controls for the Masson trichrome stain and all mucin-related histochemical stains that we tested. The goblet cells of the intestine and skin stained strongly with Alcian blue, pH 2.5 (AB-2.5), mucicarmine, colloidal iron, periodic acid Schiff (PAS) or PAS-hematoxylin (PASH) and combination stains such as colloidal iron-PASH. Cell lines were not useful for evaluating histochemical stains except for PASH. The splenic stroma was a useful control for AB-2.5; however, eosin and mucin stains stained cell lines poorly, probably due to their rapid growth and associated loss of some differentiated characteristics such as production of mucins. Nevertheless, the cell lines were a critical control for immunohistochemical stains. Immunostaining of specific cell lines was consistent with the presence of markers, e.g., EGFr in DU145 cells. The cell lines expressed a

  5. Gram's stain of peritoneal fluid is rarely helpful in the evaluation of the ascites patient.

    Science.gov (United States)

    Chinnock, Brian; Fox, Casey; Hendey, Gregory W

    2009-07-01

    We determine the sensitivity, specificity, and clinical utility of the Gram's stain of peritoneal fluid in patients undergoing paracentesis. We conducted a retrospective chart review of all peritoneal fluid analyses in a 3-year period in an urban 3-hospital system. Peritoneal dialysis and diagnostic peritoneal lavage patients were excluded. Data collected included Gram's stain, cell count, and culture results. Spontaneous bacterial peritonitis was defined as an absolute neutrophil count greater than 250 cells/mm(3). In patients with a positive Gram's stain result, charts were reviewed for antibiotic changes. Primary outcome measures were the sensitivity, specificity, and predictive values of the Gram's stain for the detection of spontaneous bacterial peritonitis. Of 796 fluid samples, Gram's stain demonstrated an organism in 31 (3.9%; 95% confidence interval [CI] 2.6% to 5.4%). Gram stain had a sensitivity of 10% (95% CI 6% to 15%), specificity of 97.5% (95% CI 96.7% to 98.3%), positive predictive value of 48% (95% CI 32% to 65%), and negative predictive value of 81.3% (95% CI 80.7% to 82.0%) in the detection of spontaneous bacterial peritonitis. Antibiotic treatment of spontaneous bacterial peritonitis was changed after Gram's stain results in only 1 case, and 16 of 31 positive Gram's stain results occurred in patients without spontaneous bacterial peritonitis, likely representing contaminants. The Gram's stain result was rarely positive in patients undergoing paracentesis, and when positive, it rarely provided clinically useful information.

  6. A Simple Procedure for the Evaluation of Bone Vitality by Staining with a Tetrazolium Salt

    Directory of Open Access Journals (Sweden)

    René Schiffner

    2017-07-01

    Full Text Available Presently, no intra-operative method for a direct assessment of bone vitality exists. Therefore, we set out to test the applicability of tetrazolium-based staining on bone samples. The explanted femoral heads of 37 patients were used to obtain either cancellous bone fragments or bone slices. Samples were stained with 2,3,5-triphenyl-2H-tetrazolium chloride (TTC or 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (thiazolyl blue, MTT at different times (one to twelve hours after explantation. Staining was quantified either spectrophotometrically after extraction of the dyes or by densitometric image analysis. TTC-staining of cancellous bone fragments and bone slices, respectively, indicated the detectability of vital cells in both types of samples in a window of up to six hours after explantation. Staining intensity at later time-points was indistinguishable from the staining of untreated samples or sodium azide treated samples, which represent dead cells. In contrast, MTT-staining of bone slices revealed intense unspecific staining, which obscured the evaluation of the vitality of the samples. The lack of a detectable increase of colour intensity in TTC-stained bone samples, which were treated more than six hours after explantation, corresponds to reduced fracture healing. The described simple procedure could provide a basis for an intraoperative decision by the orthopaedic surgeon.

  7. Selective staining of gastric biopsies for H. pylori does not affect detection rates or turnaround time and improves cost compared to reflexive staining.

    Science.gov (United States)

    Decker, Lauren; Routh, Joshua Keith; Snider, Jessica Sara; Hanson, Joshua Anspach

    2017-01-01

    To evaluate how reflexive versus selective H. pylori stains affect detection rates, turnaround time (TAT), and cost savings in a real life practice environment following an institutional policy change. The aforementioned parameters were evaluated in all cases in the year preceding and the year following an institutional policy change from reflexive to selective staining. 1497 patients comprised the reflexive stain (RS) group of which 228 (15.2%) were H. pylori positive. 1629 patients comprised the selective stain (SS) group of which 237 (14.5%) were H. pylori positive. There was no significant difference in H. pylori detection rates between the RS and SS groups (OR=0.95, 95% CI=0.78-1.15, p=0.59). TATs were similarly equivalent with a mean of 52.4h for the RS cohort and 53.7h for the SS cohort (p=0.344), both of which included a resident preview day. We calculated an average laboratory cost savings of $11.68 per case, which saved our department over $15,000 (37%) in the year following the policy change. Our results support a policy of selective staining for H. pylori as opposed to reflexive staining and go on to show that laboratories that change their policy can expect to generate cost savings without compromising detection rates or TAT. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Comparative analysis of methods for accurate recognition of cells through nuclei staining of Ki-67 in neuroblastoma and estrogen/progesterone status staining in breast cancer.

    Science.gov (United States)

    Markiewicz, Tomasz; Wisniewski, Piotr; Osowski, Stanislaw; Patera, Janusz; Kozlowski, Wojciech; Koktysz, Robert

    2009-02-01

    To compare 2 automatic systems for the recognition and counting of 2 different families of cells through nuclei staining: Ki-67 in neuroblastoma and estrogen/progesterone (ER/PR) status staining in breast cancer. Morphology-based segmentation strategies and the Support Vector Machine approach have been used for the accurate extraction and recognition of the cells. To achieve the highest possible accuracy, 2 specialized systems specially suited for Ki-67 and ER/PR staining have been developed. The testing set of histologic slides of Ki-67 and ER/PR staining has been assessed by our system and the results compared to the score of a human expert. The results are in good agreement. The average differences are within the acceptable limits of 10%. The main advantage of the system is its absolute repeatability of scores. The proposed computer-assisted automatic system of cell extraction and recognition through nuclei staining has confirmed sufficient accuracy for the tested images and may provide a useful tool for cell recognition and counting on the basis of histologic slides with Ki-67 and ER/PR staining.

  9. Ziehl-Neelsen staining technique can diagnose paragonimiasis.

    Directory of Open Access Journals (Sweden)

    Günther Slesak

    2011-05-01

    Full Text Available BACKGROUND: We evaluated the Ziehl-Neelsen staining (ZNS technique for the diagnosis of paragonimiasis in Laos and compared different modifications of the ZNS techniques. METHODOLOGY: WE APPLIED THE FOLLOWING APPROACH: We (1 examined a paragonimiasis index case's sputum with wet film direct examination (WF and ZNS; (2 re-examined stored ZNS slides from two provinces; (3 compared prospectively WF, ZNS, and formalin-ether concentration technique (FECT for sputum examination of patients with chronic cough; and (4 compared different ZNS procedures. Finally, we assessed excess direct costs associated with the use of different diagnostic techniques. PRINCIPAL FINDINGS: Paragonimus eggs were clearly visible in WF and ZNS sputum samples of the index case. They appeared brownish-reddish in ZNS and were detected in 6 of 263 archived ZNS slides corresponding to 5 patients. One hundred sputum samples from 43 patients were examined with three techniques, which revealed that 6 patients had paragonimiasis (13 positive samples. Sensitivity per slide of the FECT, ZNS and the WF technique was 84.6 (p = 0.48, 76.9 (p = 0.25 and 61.5% (p = 0.07, respectively. Percentage of fragmented eggs was below 19% and did not differ between techniques (p = 0.13. Additional operational costs per slide were 0 (ZNS, 0.10 US$ (WF, and 0.79 US$ (FECT. ZNS heated for five minutes contained less eggs than briefly heated slides (29 eggs per slide [eps] vs. 42 eps, p = 0.01. Bloodstained sputum portions contained more eggs than unstained parts (3.3 eps vs. 0.7 eps, p = 0.016. CONCLUSIONS/SIGNIFICANCE: Paragonimus eggs can easily be detected in today's widely used ZNS of sputum slides. The ZNS technique appears superior to the standard WF sputum examination for paragonimiasis and eliminates the risk of tuberculosis transmission. Our findings suggest that ZNS sputum slides should also be examined routinely for Paragonimus eggs. ZNS technique has potential in epidemiological research on

  10. Modified PAS stain: A new diagnostic method for onychomycosis.

    Science.gov (United States)

    Hajar, Tamar; Fernández-Martínez, Ramon; Moreno-Coutiño, Gabriela; Vásquez Del Mercado, Elsa; Arenas, Roberto

    2016-01-01

    Onychomycosis is the most common nail disease and represents around 50% of nail disorders. Accurate diagnosis with adequate evidence is ideal before starting any treatment. Current diagnostic methods offer low specificity and sensitivity. To create a new method for the diagnosis of onychomycosis, and to compare its sensitivity and specificity with the existing methods. One hundred and ninety-two samples with clinical suspicion of onychomycosis were included and underwent modified PAS stain (M-PAS), KOH/chlorazol black (KOH/CB) and culture testing. Sensitivity, specificity, positive and negative predictive values were calculated. In 152 out of 192 samples (79.2%) fungi structures were found in at least one of the three tests performed, and the patients were diagnosed with onychomycosis; 40 samples out of 192 (20.8%) were negative. Using M-PAS, filaments and/or spores were seen in 143 samples from the 152 positive (94%); 39 of them were negative to KOH/CB and positive to M-PAS (25.6%). With KOH/CB, filaments and/or spores were seen in 113 cases from the 152 positive samples (73.8% of the onychomycosis cases). Thirty-five cultures were positive, of which 77% were identified as Trichophyton rubrum; 117 onychomycosis cases were diagnosed despite the negative culture (76.9%). M-PAS showed 92.5% sensitivity and 55.55% specificity, a 67.5% positive predictive value and a 81.6% negative productive value. This procedure, a combination of the existing methods to diagnose onychomycosis, KOH/CB together with a nail clipping biopsy, proved to have high sensitivity, as well as being rapid, easy, inexpensive and readily available in most hospital settings. M-PAS allowed us to diagnose 39 cases (25.6% of the cases of onychomycosis) that were false negative using only KOH/CB and culture. Copyright © 2014 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  11. Staining of proteins in gels with Coomassie G-250 without organic solvent and acetic acid.

    Science.gov (United States)

    Lawrence, Ann-Marie; Besir, H Uuml Seyin

    2009-08-14

    In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents (Methanol, Ethanol or 2-Propanol) and acetic acid are used for fixation, staining and destaining of proteins in a gel after SDS-PAGE. To speed up the procedure, heating the staining solution in the microwave oven for a short time is frequently used. This usually results in evaporation of toxic or hazardous Methanol, Ethanol or 2-Propanol and a strong smell of acetic acid in the lab which should be avoided due to safety considerations. In a protocol originally published in two patent applications by E.M. Wondrak (US2001046709 (A1), US6319720 (B1)), an alternative composition of the staining solution is described in which no organic solvent or acid is used. The CBB is dissolved in bidistilled water (60-80 mg of CBB G-250 per liter) and 35 mM HCl is added as the only other compound in the staining solution. The CBB staining of the gel is done after SDS-PAGE and thorough washing of the gel in bidistilled water. By heating the gel during the washing and staining steps, the process can be finished faster and no toxic or hazardous compounds are evaporating. The staining of proteins occurs already within 1 minute after heating the gel in staining solution and is fully developed after 15-30 min with a slightly blue background that is destained completely by prolonged washing of the stained gel in bidistilled water, without affecting the stained protein bands.

  12. Photoacoustic image patterns of breast carcinoma and comparisons with Magnetic Resonance Imaging and vascular stained histopathology

    Science.gov (United States)

    Heijblom, M.; Piras, D.; Brinkhuis, M.; van Hespen, J. C. G.; van den Engh, F. M.; van der Schaaf, M.; Klaase, J. M.; van Leeuwen, T. G.; Steenbergen, W.; Manohar, S.

    2015-07-01

    Photoacoustic (optoacoustic) imaging can visualize vasculature deep in tissue using the high contrast of hemoglobin to light, with the high-resolution possible with ultrasound detection. Since angiogenesis, one of the hallmarks of cancer, leads to increased vascularity, photoacoustics holds promise in imaging breast cancer as shown in proof-of-principle studies. Here for the first time, we investigate if there are specific photoacoustic appearances of breast malignancies which can be related to the tumor vascularity, using an upgraded research imaging system, the Twente Photoacoustic Mammoscope. In addition to comparisons with x-ray and ultrasound images, in subsets of cases the photoacoustic images were compared with MR images, and with vascular staining in histopathology. We were able to identify lesions in suspect breasts at the expected locations in 28 of 29 cases. We discovered generally three types of photoacoustic appearances reminiscent of contrast enhancement types reported in MR imaging of breast malignancies, and first insights were gained into the relationship with tumor vascularity.

  13. Photoacoustic image patterns of breast carcinoma and comparisons with Magnetic Resonance Imaging and vascular stained histopathology

    Science.gov (United States)

    Heijblom, M.; Piras, D.; Brinkhuis, M.; van Hespen, J. C. G.; van den Engh, F. M.; van der Schaaf, M.; Klaase, J. M.; van Leeuwen, T. G.; Steenbergen, W.; Manohar, S.

    2015-01-01

    Photoacoustic (optoacoustic) imaging can visualize vasculature deep in tissue using the high contrast of hemoglobin to light, with the high-resolution possible with ultrasound detection. Since angiogenesis, one of the hallmarks of cancer, leads to increased vascularity, photoacoustics holds promise in imaging breast cancer as shown in proof-of-principle studies. Here for the first time, we investigate if there are specific photoacoustic appearances of breast malignancies which can be related to the tumor vascularity, using an upgraded research imaging system, the Twente Photoacoustic Mammoscope. In addition to comparisons with x-ray and ultrasound images, in subsets of cases the photoacoustic images were compared with MR images, and with vascular staining in histopathology. We were able to identify lesions in suspect breasts at the expected locations in 28 of 29 cases. We discovered generally three types of photoacoustic appearances reminiscent of contrast enhancement types reported in MR imaging of breast malignancies, and first insights were gained into the relationship with tumor vascularity. PMID:26159440

  14. β-Amyloid precursor protein staining of the brain in sudden infant and early childhood death.

    Science.gov (United States)

    Jensen, Lisbeth Lund; Banner, Jytte; Ulhøi, Benedicte Parm; Byard, Roger W

    2014-06-01

    To develop and validate a scoring method for assessing β-amyloid precursor protein (APP) staining in cerebral white matter and to investigate the occurrence, amount and deposition pattern based on the cause of death in infants and young children. Archival cerebral tissue was examined from a total of 176 cases (0 to 3 years of age). Each of the APP-stained sections was graded according to a simple scoring system based on the number and type of changes in eight anatomical regions. Examination of the sections revealed some degree of APP staining in 95% of the cases. The highest mean APP scores were found in cases of head trauma, and the lowest scores were found in the cases of drowning. APP staining, although sometimes minimal, was found in all 48 cases of and sudden infant death syndrome (SIDS). Patterns of APP staining (the amount and distribution) were different in cases of head trauma, infection and SIDS but were similar in the SIDS and asphyxia groups. This study demonstrates the use of an integrated scoring system that was developed to assess APP staining in the brain. APP staining was seen in a high proportion of cases, including relatively sudden deaths. The amount of APP was significantly higher in cases of trauma than in nontraumatic deaths. However, APP was detected within all groups. The pattern of APP staining was similar in infants who had died of SIDS and from mechanical asphyxia. © 2013 British Neuropathological Society.

  15. Application of Digital Image Analysis and Flow Cytometry To Enumerate Marine Viruses Stained with SYBR Gold†

    Science.gov (United States)

    Chen, Feng; Lu, Jing-rang; Binder, Brian J.; Liu, Ying-chun; Hodson, Robert E.

    2001-01-01

    A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of SYBR Gold-stained viruses was about twice that of SYBR Green I-stained viruses. Digital images of SYBR Gold-stained viral particles were processed to enumerate the concentration of viral particles by using digital image analysis software. Estimates of viral concentration based on digitized images were 1.3 times higher than those based on direct counting by epifluorescence microscopy. Direct epifluorescence counts of SYBR Gold-stained viral particles were in turn about 1.34 times higher than those estimated by the transmission electron microscope method. Bacteriophage lysates stained with SYBR Gold formed a distinct population in flow cytometric signatures. Flow cytometric analysis revealed at least four viral subpopulations for a Lake Erie sample and two subpopulations for a Georgia coastal sample. Flow cytometry-based viral counts for various types of samples averaged 1.1 times higher than direct epifluorescence microscopic counts. The potential application of digital image analysis and flow cytometry for rapid and accurate measurement of viral abundance in aquatic environments is discussed. PMID:11157214

  16. Comparative stain removal properties of four commercially available denture cleaning products: an in vitro study.

    Science.gov (United States)

    Alam, M; Jagger, R; Vowles, R; Moran, J

    2011-02-01

    Formulations of commercially available denture cleaners vary widely. Unfortunately, comparative data to suggest which products are the most effective can become invalid as newer products are introduced or formulations are changed. The aim of this in vitro study was to measure the stain removal properties of four currently available denture cleaners. Stain was deposited on multi-well polystyrene saliva coated microplates using multiple chlorhexidine and tea solutions. Following drying, each stained well was exposed to a solution of denture cleaner, dried again and the amount of stain remaining measured using a microplate reader. The cleaning procedure was repeated with further multiple exposures of the wells to solutions of the denture cleaners. All denture cleaners removed stain better than water used as a control. At five cleaning cycles only one of the cleaners (Superdrug Cleaning Powder) had removed 100% of the stain. At 30 cycles three of the cleaners had removed 100% of the stain. All the commercial denture cleaners removed stain. Superdrug Cleaning Powder, which contains sodium percarbonate and sodium lauryl sulphate, was particularly effective. © 2010 John Wiley & Sons A/S.

  17. Masson trichrome stain helps differentiate myofibroma from smooth muscle lesions in the head and neck region.

    Science.gov (United States)

    Chang, Julia Yu Fong; Kessler, Harvey P

    2008-10-01

    Myofibromas are well described in the head and neck region, but differentiating them from smooth muscle lesions is still difficult using smooth muscle immunohistochemical stains. This study evaluated the usefulness of the Masson trichrome stain in differentiating myofibromas from smooth muscle lesions in the head and neck region. Samples of 11 oral myofibromas, two leiomyomas, one angioleiomyoma, and one smooth muscle hamartoma were retrieved from our archives. Immunohistochemistry and Masson trichrome stains were performed on tissue sections of these lesions. All 11 oral myofibromas, seven from male patients and four from female patients, were solitary myofibromas. The patients' mean age at diagnosis was 32.8 years. Oral myofibromas occurred most commonly on the gingiva (four cases) and in the mandible (three cases). With the Masson trichrome stain, the smooth muscle cell cytoplasm was stained red, while the collagenous fibrous tissue was stained blue. Myofibromas and smooth muscle lesions demonstrated different characteristic patterns with the Masson trichrome stain. Myofibromas were composed of a much more collagenous stroma intermixed with the spindle cells. Thick fibrous bundles with random, irregularly intersecting angles were prominent in myofibromas. Smooth muscle lesions showed only minimal delicate fibrous tissue surrounding the smooth muscle cells and in the septa between the smooth muscle masses. On low-power view, red masses of smooth muscle tumor surrounded by blue fibrous tissue were observed. The Masson trichrome stain can be a useful tool to differentiate myofibromas from smooth muscle lesions, but immunohistochemical methods to rule out other spindle cell lesions are still needed.

  18. Evidence from dithizone and selenium zinc histochemistry that perivascular mossy fiber boutons stain preferentially "in vivo".

    Science.gov (United States)

    Howell, G A; Frederickson, C J; Danscher, G

    1989-01-01

    This paper describes a perivascular staining pattern that is obtained when dithizone or sodium selenite are used to label zinc intravitally. Our observations indicate that the perivascular staining is a result of zinc labeling in mossy fiber boutons adjacent to capillaries and suggest that there might be a special blood brain barrier in the mossy fiber regions.

  19. Standardization and standards for dyes and stains used in biology and medicine

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2007-01-01

    The reasons for standardization and the preparation of standards for dyes and stains are presented. The national, regional and international standardization agencies are described in detail prior to a consideration of why standards should be prepared for the field of biomedical staining. An outli...

  20. Red alder kitchen cabinets—How does application of commercial stains influence customer choice?

    Science.gov (United States)

    David Nicholls; Joseph. Roos

    2007-01-01

    A better understanding of consumer reaction and preferences for red alder (Alnus rubra Bong.) secondary products will help Alaska producers in entering new markets. In this study, red alder kitchen cabinets were commercially stained to six different levels and displayed at home shows in Portland, Oregon, and Anchorage, Alaska. The stains simulated...

  1. Identification and age estimation of blood stains on colored backgrounds by near infrared spectroscopy

    NARCIS (Netherlands)

    Edelman, G.; Manti, V.; Ruth, van S.M.; Leeuwen, van T.; Aalders, M.

    2012-01-01

    Non-destructive identification and subsequent age estimation of blood stains are significant steps in forensic casework. The latter can provide important information on the temporal aspects of a crime. As previously shown, visible spectroscopy of blood stains on white backgrounds can successfully be

  2. [The identification of barbituric acid derivatives in the old blood stains on textiles].

    Science.gov (United States)

    Kirichek, A V; Shabalina, A E; Rassinskaya, L A

    Thus article was designed to report a few cases of the identification of barbituric acid derivatives in the old blood stains on the clothes and other textiles. The data presented give evidence that barbiturates are capable of persisting in dry blood stains during rather a long period. The authors emphasize the necessity of mandatory control investigations to avoid obtaining the false positive results.

  3. 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 ...

  4. Use of hydrogen peroxide-egg albumin to eliminate nonspecific staining in immunoperoxidase techniques.

    Science.gov (United States)

    Zehr, D R

    1978-05-01

    The application of hydrogen peroxide and egg albumin to paraffin sections before immunostaining prevents nonspecific staining by immunoperoxidase techniques. This method is more effective than pretreating secions with normal sera, or using either egg albumin or hydrogen peroxide separately, or using diluted antisera with prolonged incuabations in the staining procedure.

  5. Improved method for silver staining of glycoproteins in thin sodium dodecyl sulfate polyacrylamide gels

    DEFF Research Database (Denmark)

    Møller, H J; Poulsen, J H

    1995-01-01

    A method for detection of glycoproteins in thin sodium dodecyl sulfate polyacrylamide gels was developed by a combination of (i) initial periodic acid oxidation/Alcian blue staining and (ii) subsequent staining with silver nitrate. The procedure allowed detection of as little as 1.6 ng of alpha 1...

  6. Differentiating neurotized melanocytic nevi from neurofibromas using Melan-A (MART-1) immunohistochemical stain.

    Science.gov (United States)

    Chen, Yumei; Klonowski, Paul W; Lind, Anne C; Lu, Dongsi

    2012-07-01

    Neurotized melanocytic nevi and neurofibromas are common, benign cutaneous neoplasms. Usually they are histologically distinct from each other; however, neurotized melanocytic nevi and neurofibromas can be clinically and histologically similar. To determine whether Melan-A (MART-1) immunohistochemical stain is sufficient to differentiate neurotized melanocytic nevi from neurofibromas. Forty-nine consecutive specimens of melanocytic nevi with neurotization and 49 specimens of neurofibromas were selected. We used antibodies against Melan-A, S100, and neurofilament protein. All of the melanocytic nevi showed Melan-A staining within the neurotized areas, with most of the areas staining strongly positive, whereas all the neurofibromas were completely absent of Melan-A stain. All of the nevi, including the neurotized areas, stained strongly and diffusely for S100, whereas all the neurofibromas showed a distinctive, sharp, wavy pattern of S100 staining. Neurofilament protein showed scattered staining of both melanocytic nevi and neurofibromas. Our data indicate that Melan-A immunohistochemical staining is helpful in differentiating neurotized melanocytic nevi from neurofibromas when distinction on histomorphology alone is difficult.

  7. Stained glass painting during the second half of the 20th century in Latvia, its tendency development in nowadays

    OpenAIRE

    Groza, Vineta

    2012-01-01

    Stained glass painting during the second half of the 20th century in Latvia, its tendency development in nowadays. The stained glass panel ranks among the most interesting fields of monumental decorative art. In Soviet glass art history Latvian stained glass artists occupy the foremost places in this technique. The classical stained glass technique was combined with painting and sandblasting elements, which were incorporated into geometric monumental compositions. Latvian stained glass artist...

  8. Automated differential staining for cartilage and bone in whole mount preparations of vertebrates.

    Science.gov (United States)

    Rousseaux, C G

    1985-09-01

    An automated, rapid procedure for differential staining of cartilage and bone of vertebrates is described. The process involves rapid, complete staining of freshly skinned, eviscerated specimens after 30 sec immersion in a 70 C water bath, fixation in formol acetic alcohol and a rinse in 70% alcohol. Using an automatic tissue processor, the specimen is stained in alcian blue for 24 hr and macerated in 3% potassium hydroxide for 8 hr. Staining in alizarin red with maceration in 3% potassium hydroxide is completed manually. The specimens are cleared and preserved in glycerol. Good quality evenly stained specimens can be examined in less than three days and up to 600 fetuses can be processed in less than five days.

  9. [Influence of staining and mounting methods on Cryptomeria japonica and Cupressaceae pollen counts].

    Science.gov (United States)

    Nakayama, T; Nakayama, H

    2001-07-01

    The staining and mounting method for measuring air-borne pollen differs at each institute resulting in discrepancies. We examined influence of staining and mounting methods on pollen counts of Cryptomeria japonica and Cupressaceae. Two Durham type pollen collection instruments, stored at the same place and holding slides coated with white vaseline, were exposed to the air for 24 hours. Pollen was counted using Calberla staining (C method) or gentiana-violet-glycerin jelly staining (G method). Results showed: 1) C method showed more variety than G method for measuring the pollen counts from the beginning to the end of the pollen season; 2) a significant coincidence was observed between counts measured by C and G methods (p pollen counts and that pollen count reports should include the collection and staining methods.

  10. Uncovering the origin of the black stains in Lascaux Cave in France.

    Science.gov (United States)

    Saiz-Jimenez, Cesareo; Miller, Ana Z; Martin-Sanchez, Pedro M; Hernandez-Marine, Mariona

    2012-12-01

    Lascaux Cave in France was discovered in 1940. Since being opened to visitors the cave has suffered three major microbial outbreaks. The current problem is the fast dissemination of black stains which are threatening the Palaeolithic paintings. Previous data pointed to the involvement of new fungal species in the formation of black stains on the rock walls and ceiling. However, it appears that there could be other reasons for the formation of different and extensive black stains coating the surface of the clayey sediments. Our analyses reveal that black stains on clayey sediments are mainly produced by Acremonium nepalense, a manganese oxide-depositing fungus, widely distributed in the cave. Thus, in Lascaux Cave, the black stains have a dual origin: on limestone rocks they are mainly produced by the accumulation of fungal melanins, and on clayey sediments by the biogenic deposition of black manganese oxides. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  11. Proposals for best-quality immunohistochemical staining of paraffin-embedded brain tissue slides in forensics.

    Science.gov (United States)

    Trautz, Florian; Dreßler, Jan; Stassart, Ruth; Müller, Wolf; Ondruschka, Benjamin

    2018-01-03

    Immunohistochemistry (IHC) has become an integral part in forensic histopathology over the last decades. However, the underlying methods for IHC vary greatly depending on the institution, creating a lack of comparability. The aim of this study was to assess the optimal approach for different technical aspects of IHC, in order to improve and standardize this procedure. Therefore, qualitative results from manual and automatic IHC staining of brain samples were compared, as well as potential differences in suitability of common IHC glass slides. Further, possibilities of image digitalization and connected issues were investigated. In our study, automatic staining showed more consistent staining results, compared to manual staining procedures. Digitalization and digital post-processing facilitated direct analysis and analysis for reproducibility considerably. No differences were found for different commercially available microscopic glass slides regarding suitability of IHC brain researches, but a certain rate of tissue loss should be expected during the staining process.

  12. Picrosirius red staining: a useful tool to appraise collagen networks in normal and pathological tissues.

    Science.gov (United States)

    Lattouf, Raed; Younes, Ronald; Lutomski, Didier; Naaman, Nada; Godeau, Gaston; Senni, Karim; Changotade, Sylvie

    2014-10-01

    Specific staining of the extracellular matrix components is especially helpful in studying tissue remodeling, particularly in the case of connective tissue pathologies. As developed by Junqueira and colleagues in 1979, specific staining by Picrosirius red is one of the most important stains to study collagen networks in different tissues. Under polarized light, collagen bundles appear green, red or yellow, and are easily differentiated from the black background, thus allowing for quantitative morphometric analysis. As Junqueira and colleagues point out, many studies use color staining to differentiate collagen bundles and to specify collagen types, yet other studies report that polarized colors only reflect fiber thickness and packing. Using a simple histological example, our study illustrates the inability of Picrosirius red staining to differentiate collagen types, since the absorbed amount of polarized light by this dye strictly depends on the orientation of the collagen bundles. © The Author(s) 2014.

  13. Use of urinary gram stain for detection of urinary tract infection in infants.

    Science.gov (United States)

    Lockhart, G R; Lewander, W J; Cimini, D M; Josephson, S L; Linakis, J G

    1995-01-01

    To determine whether Gram stain of urine is more sensitive than urinalysis in detecting urinary tract infection in infants. Prospective series. Urban teaching hospital emergency department. Two hundred seven infants 6 months old or less, from whom a catheterized or suprapubically aspirated urine specimen was obtained for culture. Urinary Gram stain, culture, and urinalysis were performed. With culture results as the validating standard, the Gram stain sensitivity, specificity, and predictive values were compared with urinalysis, including leukocyte esterase, nitrite, pyuria, and bacteriuria. The prevalence of positive cultures was 8.7% (18 of 207). Gram stain had higher sensitivity than overall urinalysis (94% versus 67%, P stain appears to be more reliable than urinalysis in detecting urinary tract infection in young infants.

  14. Supravital dithizone staining in the isolation of human and rat pancreatic islets

    DEFF Research Database (Denmark)

    Hansen, W A; Christie, M R; Kahn, R

    1989-01-01

    no effect on insulin release in tissue culture, on acute responses to stimulatory glucose concentrations or on the insulin content of cells. These results suggest that dithizone staining can assist in the identification of islets from the human pancreas and may prove to be a useful tool in developing......Dithizone, a zinc chelating agent, is known to selectively stain the islets of Langerhans in the pancreas. In the present study, we have used this stain to aid the identification of islets in material obtained by collagenase digestion of human pancreas. Islets were shown to rapidly and reversibly...... stain red on incubation with dithizone solution. Tissue selected on the basis of dithizone staining was shown to contain insulin-positive cells and to accumulate insulin in the medium during a subsequent period in tissue culture. Experiments with rat islets indicated that the dithizone treatment had...

  15. Automation of a Nile red staining assay enables high throughput quantification of microalgal lipid production.

    Science.gov (United States)

    Morschett, Holger; Wiechert, Wolfgang; Oldiges, Marco

    2016-02-09

    Within the context of microalgal lipid production for biofuels and bulk chemical applications, specialized higher throughput devices for small scale parallelized cultivation are expected to boost the time efficiency of phototrophic bioprocess development. However, the increasing number of possible experiments is directly coupled to the demand for lipid quantification protocols that enable reliably measuring large sets of samples within short time and that can deal with the reduced sample volume typically generated at screening scale. To meet these demands, a dye based assay was established using a liquid handling robot to provide reproducible high throughput quantification of lipids with minimized hands-on-time. Lipid production was monitored using the fluorescent dye Nile red with dimethyl sulfoxide as solvent facilitating dye permeation. The staining kinetics of cells at different concentrations and physiological states were investigated to successfully down-scale the assay to 96 well microtiter plates. Gravimetric calibration against a well-established extractive protocol enabled absolute quantification of intracellular lipids improving precision from ±8 to ±2 % on average. Implementation into an automated liquid handling platform allows for measuring up to 48 samples within 6.5 h, reducing hands-on-time to a third compared to manual operation. Moreover, it was shown that automation enhances accuracy and precision compared to manual preparation. It was revealed that established protocols relying on optical density or cell number for biomass adjustion prior to staining may suffer from errors due to significant changes of the cells' optical and physiological properties during cultivation. Alternatively, the biovolume was used as a measure for biomass concentration so that errors from morphological changes can be excluded. The newly established assay proved to be applicable for absolute quantification of algal lipids avoiding limitations of currently established

  16. [Comparison of four different staining methods for ear cytology of dogs with otitis externa].

    Science.gov (United States)

    Bouassiba, C; Osthold, W; Mueller, R S

    2013-01-01

    Cytological examination is crucial for the diagnosis and classification of canine otitis externa. Staining should reveal micro-organisms as perpetuating factors of otitis externa. The aim of the study was to compare four different staining methods (Diff-Quik®, Diff-Quik® after dipping in acetone, Gram Quick stain® and a commercial rapid stain for otitis externa) for ear cytology of dogs with otitis externa and to investigate the agreement of cytology and culture. In a study evaluating dogs with otitis externa, five ear swabs (one for culture and four for cytology) were taken from the horizontal part of the external auditory canal of 224 affected ears and compared semi-quantitatively. Diff-Quik® with and without prior dipping in acetone as well as the Gram Quick stain® displayed a high degree of agreement in the detection of micro-organisms (cocci p = 0.2366; rods p = 0.4832; yeasts p = 0.1574), while the commercial otitis rapid stain revealed significantly less micro-organisms (p   70%; the agreement was lower with the commercial otitis rapid stain. The quickest and easiest method was staining with Diff-Quik®. Diff-Quik® with or without prior dipping in acetone and the Gram Quick stain® had a high agreement in the detection of microorganisms and can thus be considered nearly equivalent for the diagnosis of otitis externa infectiosa. The commercial otitis rapid stain is less reliable. Based on this study Diff-Quik® can be recommended for the routine cytology of ear swabs. Additionally, a culture may be indicated and must be interpreted in the context of the cytology.

  17. A Randomized Controlled Clinical Trial to Evaluate Extrinsic Stain Removal of a Whitening Dentifrice.

    Science.gov (United States)

    Terézhalmy, Géza; He, Tao; Anastasia, Mary Kay; Eusebio, Rachelle

    2016-12-01

    To evaluate the extrinsic stain removal efficacy of a new whitening dentifrice containing sodium hexametaphosphate (SHMP) over a two-week period. This study used a controlled and randomized, examiner-blind, single-center, two-treatment, parallel group design. Subjects with visible extrinsic dental stain on facial surfaces of their anterior teeth, and meeting all study criteria, were entered into the trial. The test group received the whitening dentifrice with sodium fluoride and SHMP and an ADA reference soft manual toothbrush. Subjects in the control group received a dental prophylaxis after the initial examination at Baseline and were instructed to use their usual oral hygiene products at home. Subjects returned at Day 3 and Week 2 for re-evaluation of extrinsic dental stain. Extrinsic stain was measured using the Interproximal Modified Lobene (IML) Stain Index; safety was assessed based on clinical examination. Fifty subjects (mean age 32.0 years) completed the study, with 25 in each group. Statistically significant reductions in composite stain for whole tooth, as well as interproximal, gingival, and body surfaces were observed for both groups at Day 3 and Week 2 (p 0.3). At Day 3, median percent reductions in composite IML stain from Baseline were 98% for the prophylaxis group and 100% for the test dentifrice group. At Week 2, median percent reductions in composite IML stain were 100% compared to Baseline for both groups. No adverse events were reported for either group. The whitening dentifrice demonstrated a statistically significant reduction in IML stain after three days and two weeks of use relative to baseline. Stain reduction with the toothpaste was comparable to a dental prophylaxis.

  18. The staining pattern of brilliant blue G during macular hole surgery: a clinicopathologic study.

    Science.gov (United States)

    Steel, David H W; Dinah, Christiana; Madi, Haifa A; Magdi, Haifa; White, Kathryn; Rees, Jon

    2014-08-14

    To describe the intraoperative staining pattern of the internal limiting membrane (ILM)-specific dye Brilliant Blue G (BBG) in a cohort of patients with idiopathic macular holes; to analyze the associations of the staining pattern with pre- and postoperative variables and to correlate the staining pattern with transmission electron microscopy (TEM) of the excised ILM. Fifty-five consecutive patients were studied. The staining pattern was divided into three subtypes based on the intraoperative appearance. The presence of a narrow rim of nonstaining around the macular hole (MH) edge was noted and measured. In the final 21 patients, the excised ILM was examined with TEM. The pattern of staining observed was categorized as uniform in 33 patients (60%), patchy nonstaining in 17 (31%), and no visible staining in 5 (9%). The staining pattern correlated with the MH stage. In the patients with uniform or patchy staining, a nonstaining rim was observed in 26 (52%) of the 50. The presence of a rim was associated with a greater hole diameter and lower postoperative visual acuity. The stain pattern correlated significantly with the amount of cellular tissue on the vitreous side of the ILM on TEM, with a greater proportion of multicellular layer membranes and new collagen in the incomplete staining groups. A variety of nonstaining patterns around macular holes can be observed using BBG, and these patterns correlate to the amount of cellular tissue on the vitreous side of the ILM seen histologically. These patterns could be used to guide the ILM peeling requirement or extent in future studies. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

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

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    J. Rieger

    2013-07-01

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

  20. Automatic disease screening method using image processing for dried blood microfluidic drop stain pattern recognition.

    Science.gov (United States)

    Sikarwar, Basant S; Roy, Mukesh; Ranjan, Priya; Goyal, Ayush

    2016-07-01

    This paper examines programmed automatic recognition of infection from samples of dried stains of micro-scale drops of patient blood. This technique has the upside of being low-cost and less-intrusive and not requiring puncturing the patient with a needle for drawing blood, which is especially critical for infants and the matured. It also does not require expensive pathological blood test laboratory equipment. The method is shown in this work to be successful for ailment identification in patients suffering from tuberculosis and anaemia. Illness affects the physical properties of blood, which thus influence the samples of dried micro-scale blood drop stains. For instance, if a patient has a severe drop in platelet count, which is often the case of dengue or malaria patients, the blood's physical property of viscosity drops substantially, i.e. the blood is thinner. Thus, the blood micro-scale drop stain samples can be utilised for diagnosing maladies. This paper presents programmed automatic examination of the dried micro-scale drop blood stain designs utilising an algorithm based on pattern recognition. The samples of micro-scale blood drop stains of ordinary non-infected people are clearly recognisable as well as the samples of micro-scale blood drop stains of sick people, due to key distinguishing features. As a contextual analysis, the micro-scale blood drop stains of patients infected with tuberculosis have been contrasted with the micro-scale blood drop stains of typical normal healthy people. The paper dives into the fundamental flow mechanics behind how the samples of the dried micro-scale blood drop stain is shaped. What has been found is a thick ring like feature in the dried micro-scale blood drop stains of non-ailing people and thin shape like lines in the dried micro-scale blood drop stains of patients with anaemia or tuberculosis disease. The ring like feature at the periphery is caused by an outward stream conveying suspended particles to the edge

  1. Automation-assisted cervical cancer screening in manual liquid-based cytology with hematoxylin and eosin staining.

    Science.gov (United States)

    Zhang, Ling; Kong, Hui; Ting Chin, Chien; Liu, Shaoxiong; Fan, Xinmin; Wang, Tianfu; Chen, Siping

    2014-03-01

    Current automation-assisted technologies for screening cervical cancer mainly rely on automated liquid-based cytology slides with proprietary stain. This is not a cost-efficient approach to be utilized in developing countries. In this article, we propose the first automation-assisted system to screen cervical cancer in manual liquid-based cytology (MLBC) slides with hematoxylin and eosin (H&E) stain, which is inexpensive and more applicable in developing countries. This system consists of three main modules: image acquisition, cell segmentation, and cell classification. First, an autofocusing scheme is proposed to find the global maximum of the focus curve by iteratively comparing image qualities of specific locations. On the autofocused images, the multiway graph cut (GC) is performed globally on the a* channel enhanced image to obtain cytoplasm segmentation. The nuclei, especially abnormal nuclei, are robustly segmented by using GC adaptively and locally. Two concave-based approaches are integrated to split the touching nuclei. To classify the segmented cells, features are selected and preprocessed to improve the sensitivity, and contextual and cytoplasm information are introduced to improve the specificity. Experiments on 26 consecutive image stacks demonstrated that the dynamic autofocusing accuracy was 2.06 μm. On 21 cervical cell images with nonideal imaging condition and pathology, our segmentation method achieved a 93% accuracy for cytoplasm, and a 87.3% F-measure for nuclei, both outperformed state of the art works in terms of accuracy. Additional clinical trials showed that both the sensitivity (88.1%) and the specificity (100%) of our system are satisfyingly high. These results proved the feasibility of automation-assisted cervical cancer screening in MLBC slides with H&E stain, which is highly desirable in community health centers and small hospitals. © 2013 International Society for Advancement of Cytometry.

  2. A laboratory investigation of stain removal from enamel surface: comparative efficacy of three electric toothbrushes.

    Science.gov (United States)

    Schemehorn, B R; Henry, G M

    1996-07-01

    To evaluate in a controlled laboratory investigation the efficacy of three electric toothbrushes with respect to removal of extrinsic dental stain from labial enamel surfaces of bovine permanent central incisors. Enamel specimens were treated to produce a uniformly stained pellicle film. Staining was assessed photometrically, before and after brushing 16 specimens per group for 4.5 minutes with either the Braun Oral-B Plaque Remover (D7), the Braun Oral-B Ultra Plaque Remover (D9) or a high frequency electric toothbrush. A manual brush with the ADA reference abrasive was used as a positive control, and this was compared with the three electric toothbrushes, using a dentifrice slurry prepared from Crest toothpaste. Tension on the enamel specimens was set at 150 g, except for the high frequency toothbrush which was set at the manufacturer's recommended tension of 50 g. Stain removal was expressed as a cleaning ratio (CR), where larger values represented greater removal of stained pellicle. Compared with the ADA control (CR = 100), the D9 was the most effective (CR = 98) removing significantly more stain (P toothbrush was found to be the least effective (CR = 48), removing significantly less stain than any of the other toothbrushes (P < 0.05).

  3. 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.

  4. A procedure for Alcian blue staining of mucins on polyvinylidene difluoride membranes.

    Science.gov (United States)

    Dong, Weijie; Matsuno, Yu-ki; Kameyama, Akihiko

    2012-10-16

    The isolation and characterization of mucins are critically important for obtaining insight into the molecular pathology of various diseases, including cancers and cystic fibrosis. Recently, we developed a novel membrane electrophoretic method, supported molecular matrix electrophoresis (SMME), which separates mucins on a polyvinylidene difluoride (PVDF) membrane impregnated with a hydrophilic polymer. Alcian blue staining is widely used to visualize mucopolysaccharides and acidic mucins on both blotted membranes and SMME membranes; however, this method cannot be used to stain mucins with a low acidic glycan content. Meanwhile, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, but is incompatible with glycan analysis, which is indispensable for mucin characterizations. Here we describe a novel staining method, designated succinylation-Alcian blue staining, for visualizing mucins on a PVDF membrane. This method can visualize mucins regardless of the acidic residue content and shows a sensitivity 2-fold higher than that of Pro-Q Emerald 488, a fluorescent periodate Schiff-base stain. Furthermore, we demonstrate the compatibility of this novel staining procedure with glycan analysis using porcine gastric mucin as a model mucin.

  5. Rapid alkaline methylene blue supravital staining for assessment of anterior segment infections.

    Science.gov (United States)

    Kiuchi, Katsuji

    2016-01-01

    To present the Löffler's alkaline methylene blue technique of staining eye discharges in eyes with anterior segment infections. The Löffler's alkaline methylene blue staining method is a simple staining technique that can be used to differentiate bacterial, viral, and fungal infections. It is a cationic dye that stains cells blue because the positively charged dye is attracted to negatively charged particles such as polyphosphates, DNAs, and RNAs. Specimens collected from patients by swabbing are smeared onto microscope slides and the methylene blue solution is dropped on the slide. The slide is covered with a glass cover slip and examined under a microscope. The entire time from the collection to the viewing is about 30 seconds. Histopathological images of the conjunctival epithelial cells and neutrophils in eye discharges were dyed blue and the nuclei were stained more intensely blue. Bacterial infections consisted mainly of neutrophils, and viral infections consisted mainly of lymphocytes. Löffler's alkaline methylene blue staining can be done in about 30 seconds for diagnosis. Even though this is a one color stain, it is possible to infer the cause of the infection by detection of the absence of bacteria and/or fungi in context of the differential distribution of neutrophils and lymphocytes.

  6. Abelardo Gallego (1879-1930) and his contributions to histotechnology: the Gallego stains.

    Science.gov (United States)

    Ortiz-Hidalgo, Carlos

    2011-02-01

    The Gallego general tissue stain is used in histology and histopathology and stains beautifully connective tissue, in which all nuclei are stained in magenta red, epithelial cytoplasm in red-yellow, connective tissue is stained in brilliant green, muscle in olive green, and keratinized epithelium and blood in grass green. There is also Gallego's method for differential staining of elastic tissue that enables a complete differentiation between elastic and collagenous connective tissue. The elastic tissue is stained in brilliant fuschin red, while collagen is stained in brilliant green. Abelardo Gallego was born in Spain on September 10, 1879. He was appointed as professor of Pharmacology and Therapeutics at the Faculty of Veterinary Medicine at the University of Santiago de Compostela, Spain, and in 1921 became full-time professor of Histology and Pathology in Madrid supported by Ramón y Cajal. Gallego conducted extensive research into veterinary pathology. Don Abelardo was a quiet, reserved, but friendly, sensitive, optimistic and persistent person. Weakened by a bout of bronchitis, Gallego died aged 51 on February 3, 1930, and was buried at the Almudena Cemetery, in Madrid. Gallego was one of the most outstanding scientists of Spanish veterinary medicine and some of his achievements are reviewed. Copyright © 2009 Elsevier GmbH. All rights reserved.

  7. Improved coomassie blue dye-based fast staining protocol for proteins separated by SDS-PAGE.

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    Pavel Májek

    Full Text Available The time required to visualize proteins using Coomassie Blue dye has been significantly reduced with the introduction of fast staining protocols based on staining with a Coomassie Blue dye solution at boiling temperatures. However, fast stainings suffer from high gel backgrounds, reducing the signal-to-noise ratio and limiting the number of detectable spots in the case of 2D SDS-PAGE. The aim of this work was to eliminate the high gel background, and thus improve fast staining protocols based on Coomassie Blue dye. We show that merely replacing water with a 4 mM EDTA washing solution at boiling temperatures, results in a transparent gel background within 50 to 60 minutes of destaining. Moreover, when a combination of imidazole-zinc reverse staining and Coomassie Blue-based fast staining is used the sensitivity is improved significantly; nanogram amounts of proteins can be detected using 1D SDS-PAGE, and about 30% to 60% more spots can be detected with 2D SDS-PAGE in plasma, platelet, and rat brain tissue samples. This work represents an optimized fast staining protocol with improved sensitivity, requiring between 60 to 75 minutes to complete protein visualization.

  8. Improving Gram stain proficiency in hospital and satellite laboratories that do not have microbiology.

    Science.gov (United States)

    Guarner, Jeannette; Street, Cassandra; Matlock, Margaret; Cole, Lisa; Brierre, Francoise

    2017-03-01

    Consolidation of laboratories has left many hospitals and satellite laboratories with minimal microbiologic testing. In many hospitals and satellite laboratories, Gram stains on primary specimens are still performed despite difficultly in maintaining proficiency. To maintain Gram stain proficiency at a community 450-bed hospital with an active emergency room we designed bimonthly challenges that require reporting Gram staining and morphology of different organisms. The challenges consist of five specimens prepared by the reference microbiology laboratory from cultures and primary specimens. Twenty to 23 medical laboratory scientists participate reading the challenges. Results from the challenges are discussed with each medical laboratory scientists. In addition, printed images from the challenges are presented at huddle to add microbiology knowledge. On the first three challenges, Gram staining was read correctly in 71%-77% of the time while morphology 53%-66%. In the last six challenges correct answers for Gram stain were 77%-99% while morphology 73%-96%. We observed statistically significant improvement when reading Gram stains by providing frequent challenges to medical laboratory scientists. The clinical importance of Gram stain results is emphasized during huddle presentations increasing knowledge and motivation to perform the test for patients.

  9. 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

  10. The role of serum amyloid A staining of granulomatous tissues for the diagnosis of sarcoidosis.

    Science.gov (United States)

    Huho, Albert; Foulke, Llewellyn; Jennings, Timothy; Koutroumpakis, Efstratios; Dalvi, Siddhartha; Chaudhry, Haroon; Chopra, Amit; Modi, Aakash; Rane, Neha; Prezant, David J; Sheehan, Christine; Yucel, Recai; Patel, Mehul; Judson, Marc A

    2017-05-01

    Previous studies demonstrated that SAA staining of sarcoidosis granulomas was qualitatively and quantitatively different from other granulomatous diseases. These data suggest that positive SAA staining of granulomatous tissue may have adequate specificity to establish a diagnosis of sarcoidosis. Our objective was to determine the diagnostic specificity of SAA staining for sarcoidosis relative to other granulomatous disorders. Pathological specimens demonstrating granulomatous inflammation were retrospectively identified at one institution, plus 4 specimens were obtained from New York City firefighters with biopsy-confirmed World Trade Center "sarcoidosis-like" pulmonary disease. Specimens were analyzed if specific diagnoses related to the granulomatous inflammation were confirmed through medical record review. SAA staining was performed using previously developed methods. Two pathologists, blinded to each other and the diagnoses, determined if the stained material was SAA positive or negative. Discordant results were adjudicated by the two pathologists. 106 specimens were analyzed from 100 patients, with 36 biopsies (34%) from sarcoidosis tissues and 70 (66%) from other granulomatous disorders. The Cohen Kappa correlation between the two pathologists for SAA staining positivity was excellent (0.85, 0.73-0.98). The overall specificity of SAA staining for the diagnosis of sarcoidosis was 84% (59/70). The sensitivity was 44% (16/36). Although SAA staining of various granulomatous tissues was fairly specific for the diagnosis of sarcoidosis, the specificity was inadequate for SAA staining to be used as a diagnostic test for sarcoidosis in isolation. These data suggest that SAA production may not be a universal mechanism in the development of sarcoidosis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. 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.

  12. Assessing Nezara viridula (Hemiptera: Pentatomidae) feeding damage in macadamia nuts by using a biological stain.

    Science.gov (United States)

    Golden, Mary; Follett, Peter A; Wright, Mark G

    2006-06-01

    Damage caused by southern green stink bug, Nezara viridula (L.), to macadamia nuts, Macadamia integrifolia Maiden & Betche, is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in macadamia nuts. By using the staining method, feeding probes were easily detected on the husk, shell, and kernel. Husk probing was highly correlated (0.80-0.90) with feeding and damage to the kernel. Failure rate to detect kernel damage from stained husk probes was generally management tactics and forecast outbreak populations.

  13. Amazonian açai and food dyes for staining arbuscular- micorrhizal fungi

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    Aline Lourdes Martins Silva

    2015-12-01

    Full Text Available Arbuscular mycorrhizae microscopy requires differential staining of typical structures. Dyes employed, such as trypan blue, pose risks to health and environment. Alternative dyes such as pen ink and aniline have variable coloring efficiency. In this work, Brachiaria decumbens roots, discolored with caustic soda (NaOH, were stained with açai, annatto, saffron, trypan blue and pen inks. There were significant differences among dyes regarding stained mycorrhizal structures and pictures quality. Acai was considered the best alternative dye, with similar results to trypan blue.

  14. A novel staining method for quantification and 3D visualisation of capillaries and muscle fibres

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    V Cebasek

    2009-06-01

    Full Text Available The aim of this study was to introduce a combined fluorescent staining that clearly demonstrates capillaries and distinguishes them from the basal lamina of muscle fibres in skeletal muscle tissue. The triple staining with CD31, Griffonia (Bandeira simplicifolia lectin (GSL I and laminin efficiently distinguishes vascular endothelium from the basal lamina of skeletal muscle fibres in physiological and pathological conditions. The presented triple staining method has several advantages, which facilitate quantitative analysis of the capillary network, and its relation to individual muscle fibres.

  15. [An endoscopy-guided plan of radiotherapy for detecting esophageal cancer using a lugol staining].

    Science.gov (United States)

    Kobayashi, H; Itoh, T; Kato, T; Fujita, Y; Murata, R; Tanabe, M; Kinashi, Y; Ono, K; Abe, M

    1990-04-01

    The Lugol staining method has lately come to play an important role in the detection of superficial esophageal cancers which are extremely difficult to detect by using only a barium double-contrasted esophagiogram and conventional esophagioscopy. Thus we have tested an endoscopy-guided plan of radiotherapy for detecting an esophageal cancer using Lugol stain. This method was found useful in detecting intraepithelial small lesions, such as intramural metastasis or multiple origin cancers. Additionally, it has been found useful in determining the definite shape of superficial malignant lesions by evaluating biopsy specimens taken from the border of the non-stained regions.

  16. The Role of Hemiwicking on the Shape of a Blood Drop Stain

    Science.gov (United States)

    Shiri, Samira; Martin, Kenneth; Bird, James

    2017-11-01

    Blood pattern analysis (BPA) typically assumes that an elliptical stain is due to oblique drop impact. From the eccentricity of the elliptical stain - while also accounting for gravity and drag - the source and trajectory of the blood drops can be estimated. Yet, these models generally neglect any fluid motion following impact that could influence the shape of the stain. Here we demonstrate that under certain conditions on certain materials, a blood drop will undergo anisotropic hemiwicking. Through systemic experiments and modeling, we aim to better understand this phenomenon with the goal of ultimately decreasing the uncertainty in crime scene reconstruction.

  17. A system for the automatic estimation of morphometric parameters of corneal endothelium in alizarine red-stained images.

    Science.gov (United States)

    Ruggeri, Alfredo; Scarpa, Fabio; De Luca, Massimo; Meltendorf, Christian; Schroeter, Jan

    2010-05-01

    BACKGROUND/AIMS A computer program for the automatic estimation of endothelium morphometric parameters (cell density, pleomorphism, polymegethism) in alizarine red-stained images is presented and evaluated. METHODS Images of corneal endothelium from 30 porcine eyes stained with alizarine red were acquired with an optical microscope and saved as grey-level digital images. Each image was first pre-processed for luminosity correction and contrast enhancement. An artificial neural network was used to classify all pixels as cell contour or cell body pixels. The segmented cell contours were then used to obtain estimates of morphometric parameters. The central area was assessed and the mean area per cornea was 0.54+/-0.07 mm(2). The whole system was implemented as a computer program using the Matlab language. Estimated parameters were compared with the corresponding values derived from manual contour detection on the same images used for the automatic estimation. RESULTS For the 30 images in our dataset, the mean differences for automatic versus manual parameters were -12+/-52 (range -103 to +145) cells/mm(2) for density, 0.5+/-2.6% (range -5.6 to +5.6%) for pleomorphism and -0.7+/-1.9% (range -4.1 to +2.8%) for polymegethism. CONCLUSION The evaluation of the automatic system on 30 images from porcine eyes confirmed its ability to estimate reliably morphometric parameters with respect to parameter values derived by manual analysis.

  18. Comparison of methods of microvascular staining and quantification in prostate carcinoma

    DEFF Research Database (Denmark)

    Offersen, Birgitte Vrou; Borre, Michael; Sørensen, Flemming Brandt

    2002-01-01

    of formalin-fixed, paraffin-embedded tumor specimens from TURPs on 51 consecutive patients with prostate carcinoma were immunostained for CD34 and von Willebrand Factor (vWF). Estimates of microvascular density were based on projecting a 10 x 10 grid or a Chalkley grid onto a vascular hot spot of the invasive...... prostate carcinoma. Anti-CD34 antibodies stained microvessels in all 51 tumors, whereas anti-vWF antibodies in 6 tumors resulted in intense background staining causing omission of these. Anti-CD34 antibodies highlighted significantly more microvessels than anti-vWF antibodies, and the anti-CD34 vascular...... scores with either of the counting methods were significantly correlated, which was not the case with vWF. Both grids used on anti-CD34-stained sections resulted in vascular scores that could separate the tumors into prognostic groups. This was not possible using the Chalkley grid on vWF-stained tumors...

  19. 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

  20. Rapid Staining Method to Detect and Identify Downy Mildew (Peronospora belbahrii in Basil

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    Adolfina R. Koroch

    2013-07-01

    Full Text Available Premise of the study: Demand for fresh-market sweet basil continues to increase, but in 2009 a new pathogen emerged, threatening commercial field/greenhouse production and leading to high crop losses. This study describes a simple and effective staining method for rapid microscopic detection of basil downy mildew (Peronospora belbahrii from leaves of basil (Ocimum basilicum. Methods and Results: Fresh leaf sections infected with P. belbahrii were placed on a microscope slide, cleared with Visikol™, and stained with iodine solution followed by one drop of 70% sulfuric acid. Cell walls of the pathogen were stained with a distinct coloration, providing a high-contrast image between the pathogen and plant. Conclusions: This new staining method can be used successfully to identify downy mildew in basil, which then can significantly reduce its spread if identified early, coupled with mitigation strategies. This technique can facilitate the control of the disease, without expensive and specialized equipment.

  1. Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle Lesions in the Head and Neck Region

    Directory of Open Access Journals (Sweden)

    Julia Yu Fong Chang

    2008-10-01

    Conclusion: The Masson trichrome stain can be a useful tool to differentiate myofibromas from smooth muscle lesions, but immunohistochemical methods to rule out other spindle cell lesions are still needed.

  2. Might the Masson trichrome stain be considered a useful method for categorizing experimental tendon lesions?

    Science.gov (United States)

    Martinello, Tiziana; Pascoli, Francesco; Caporale, Giovanni; Perazzi, Anna; Iacopetti, Ilaria; Patruno, Marco

    2015-08-01

    Strain injuries of tendons are the most common orthopedic injuries in athletic subjects, be they equine or human. When the tendon is suddenly damaged, an acute inflammatory phase occurs whereas its repetitive overloading may cause chronic injuries. Currently the criteria used for grading injuries are general and subjective, and therefore a reliable grading method would be an improvement. The main purpose of this study was to assess qualitatively the histological pattern of Masson trichrome stain in healthy and injured tendons; indeed, the known "paradox" of Masson staining was used to create an evaluation for the matrix of tendons, following experimental lesions and natural repair processes. A statistically significant difference of aniline-staining between healthy and lesioned tendons was observed. Overall, we think that the Masson staining might be regarded as an informative tool in discerning the collagen spatial arrangement and therefore the histological characteristics of tendons.

  3. Stain- and Culture-Negative Necrotizing Granulomas in an Urban Population

    National Research Council Canada - National Science Library

    Burke, A; Paulk, A

    2016-01-01

    Context: A recent study found that 10% of necrotizing granulomas are idiopathic. We aimed to study the rate of stain-negative and culturenegative necrotizing granulomas within an urban population. Design...

  4. Electron tomography of negatively stained complex viruses: application in their diagnosis

    Directory of Open Access Journals (Sweden)

    Demeestere Lien

    2009-02-01

    Full Text Available Abstract Background Electron tomographic analysis can be combined with the simple and rapid negative staining technique used in electron microscopy based virus diagnosis. Methods Standard negative staining of representative examples of parapoxviruses and paramyxoviruses was combined with electron tomographic analysis. Results Digital sectioning of reconstructions of these viruses at a selected height demonstrated the viral ultrastructure in detail, including the characteristic diagnostic features like the surface threads on C-particles of a parapoxvirus and individual glycoproteins and the internal nucleoprotein strand of Newcastle disease virus. For both viruses, deformation and flattening were observed. Conclusion The combination of negative staining of complex viruses with electron tomographic analysis, allows visualizing and measuring artifacts typical for negative staining. This approach allows sharp visualisation of structures in a subnanometer-thick plane, avoiding blurring due to superposition which is inherent to TEM. In selected examples, such analyses can improve diagnosis of viral agents.

  5. Chemical contrast observed in thermal images of blood-stained fabrics exposed to steam.

    Science.gov (United States)

    O'Brien, Wayne L; Boltin, Nicholas D; Lu, Zhenyu; Cassidy, Brianna M; Belliveau, Raymond G; Straub, Emory J; DeJong, Stephanie A; Morgan, Stephen L; Myrick, M L

    2015-09-21

    Thermal imaging is not ordinarily a good way to visualize chemical contrast. In recent work, however, we observed strong and reproducible images with chemical contrasts on blood-stained fabrics, especially on more hydrophobic fabrics like acrylic and polyester.

  6. Staining pattern classification of antinuclear autoantibodies based on block segmentation in indirect immunofluorescence images.

    Directory of Open Access Journals (Sweden)

    Jiaqian Li

    Full Text Available Indirect immunofluorescence based on HEp-2 cell substrate is the most commonly used staining method for antinuclear autoantibodies associated with different types of autoimmune pathologies. The aim of this paper is to design an automatic system to identify the staining patterns based on block segmentation compared to the cell segmentation most used in previous research. Various feature descriptors and classifiers are tested and compared in the classification of the staining pattern of blocks and it is found that the technique of the combination of the local binary pattern and the k-nearest neighbor algorithm achieve the best performance. Relying on the results of block pattern classification, experiments on the whole images show that classifier fusion rules are able to identify the staining patterns of the whole well (specimen image with a total accuracy of about 94.62%.

  7. Efficacy of baking soda-containing chewing gum in removing natural tooth stain.

    Science.gov (United States)

    Mankodi, S M; Conforti, N; Berkowitz, H

    2001-07-01

    A 14-week, double-blind, randomized clinical trial was conducted with 126 healthy volunteers to compare the efficacy of twice-daily use of 3 baking soda-containing chewing gums in removing natural tooth stain when used in conjunction with a program of regular oral hygiene. All 3 chewing gums significantly reduced extrinsic stain (P Baking Soda Gum (AHDC) reduced dental stain by 70.8%, compared to reductions of 71.9% and 65.3%, after use of 2 experimental gum formulations. Whitened appearance improved by 1.73 shade tabs using AHDC gum, and up to 2.49 shade tabs with the experimental formulations. These results suggest that the use of baking soda-containing gum after meals, in conjunction with good oral hygiene, can improve both extrinsic dental staining and the whitened appearance of teeth.

  8. Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.

    Science.gov (United States)

    Trinh, Le A; McCutchen, Marshall D; Bonner-Fraser, Marianne; Fraser, Scott E; Bumm, Lloyd A; McCauley, David W

    2007-06-01

    In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP). The solid reaction product has significant fluorescence that enables the use of confocal microscopy to generate high-resolution three-dimensional (3-D) imaging of gene expression.

  9. Peri-stent contrast staining, major evaginations and severe malapposition after biolimus-eluting stent implantation

    DEFF Research Database (Denmark)

    Antonsen, Lisbeth; Thayssen, Per; Jensen, Lisette Okkels

    2014-01-01

    Peri-stent contrast staining and late acquired malapposition represent pathological vessel wall healing patterns following percutaneous coronary intervention with stent implantation. Earlier studies have described these abnormal vessel wall responses commonly present after implantation of first......-generation drug-eluting stents. These coronary vascular changes can cause flow disturbance and thereby dispose for later thrombotic events. This case report, based on coronary optical frequency domain imaging, describes peri-stent contrast staining, major evaginations and severe malapposition occurring 18months...

  10. Detection of Entamoeba histolytica in experimentally induced amoebic liver abscess: comparison of three staining methods

    OpenAIRE

    Tan Zi Ning; Wong Weng Kin; Shaymoli Mustafa; Arefuddin Ahmed; Rahmah Noordin; Tan Gim Cheong; Olivos-Garcia Alfonso; Lim Boon Huat

    2012-01-01

    Objective: To compare the efficacy of three different tissue stains, namely haematoxylin and eosin (H&E), periodic-acid Schiff (PAS) and immunohistochemical (IHC) stains for detection of Entamoeba histolytica (E. histolytica) trophozoites in abscessed liver tissues of hamster. Methods: Amoebic liver abscess was experimentally induced in a hamster by injecting 1 × 106 of axenically cultured virulent E. histolytica trophozoites (HM1-IMSS strain) into the portal vein. After a week post-inocul...

  11. A memory-efficient staining algorithm in 3D seismic modelling and imaging

    Science.gov (United States)

    Jia, Xiaofeng; Yang, Lu

    2017-08-01

    The staining algorithm has been proven to generate high signal-to-noise ratio (S/N) images in poorly illuminated areas in two-dimensional cases. In the staining algorithm, the stained wavefield relevant to the target area and the regular source wavefield forward propagate synchronously. Cross-correlating these two wavefields with the backward propagated receiver wavefield separately, we obtain two images: the local image of the target area and the conventional reverse time migration (RTM) image. This imaging process costs massive computer memory for wavefield storage, especially in large scale three-dimensional cases. To make the staining algorithm applicable to three-dimensional RTM, we develop a method to implement the staining algorithm in three-dimensional acoustic modelling in a standard staggered grid finite difference (FD) scheme. The implementation is adaptive to the order of spatial accuracy of the FD operator. The method can be applied to elastic, electromagnetic, and other wave equations. Taking the memory requirement into account, we adopt a random boundary condition (RBC) to backward extrapolate the receiver wavefield and reconstruct it by reverse propagation using the final wavefield snapshot only. Meanwhile, we forward simulate the stained wavefield and source wavefield simultaneously using the nearly perfectly matched layer (NPML) boundary condition. Experiments on a complex geologic model indicate that the RBC-NPML collaborative strategy not only minimizes the memory consumption but also guarantees high quality imaging results. We apply the staining algorithm to three-dimensional RTM via the proposed strategy. Numerical results show that our staining algorithm can produce high S/N images in the target areas with other structures effectively muted.

  12. B-Amyloid Precursor Protein Staining of the Brain in Sudden Infant and Early Childhood Death

    DEFF Research Database (Denmark)

    Jensen, Lisbeth Lund; Banner, Jytte; Ulhøi, Benedicte Parm

    2013-01-01

    To develop and validate a scoring method for assessing β-amyloid precursor protein (APP) staining in cerebral white matter and to investigate the occurrence, amount and deposition pattern based on the cause of death in infants and young children.......To develop and validate a scoring method for assessing β-amyloid precursor protein (APP) staining in cerebral white matter and to investigate the occurrence, amount and deposition pattern based on the cause of death in infants and young children....

  13. An evaluation of a technique to remove stains from teeth using microabrasion.

    Science.gov (United States)

    Price, Richard B T; Loney, Robert W; Doyle, M Gorman; Moulding, M Brent

    2003-08-01

    Microabrasion using a paste made of acid and pumice is a technique that has been used to remove white, yellow and brown stains from enamel. The authors evaluated the technique by studying the effectiveness of a proprietary microabrasion product. One author used microabrasion to remove white, yellow and brown stains from within the outermost layer of the tooth enamel of 32 subjects. Standardized slides of the teeth were taken before and one week after treatment. Four prosthodontists evaluated the paired images, using a standardized questionnaire and visual analog scales ranging from 1 (no improvement in appearance or stain not removed at all) to 7 (exceptional improvement in appearance or stain totally removed). The evaluators were calibrated and blinded. The evaluators always identified a difference between the pretreatment slides and posttreatment slides; they found no difference between the control slides. In all cases but one (97 percent), the treated teeth had improved in appearance with more uniformity in color. Analysis of variance revealed no differences between evaluator ratings (P = .146). The intraclass correlation coefficient for ratings of individual cases by different evaluators was 0.72, representing a "good" level of correlation of the ratings for improvement of appearance and for stain removal. Mean (+/- standard deviation) ratings were 5.38 (+/- 1.26) for improvement of appearance and 5.06 (+/- 1.26) for stain removal. This study showed that enamel microabrasion could remove stains from within the outermost layer of tooth enamel, thereby improving the appearance of the teeth. This study supports recommendations that enamel microabrasion is an effective, atraumatic method of improving the appearance of teeth with stains in the outermost layer of enamel.

  14. Measurement of stain removal in vitro: a comparison of two instrumental methods.

    Science.gov (United States)

    Lath, D L; Johnson, C; Smith, R N; Brook, A H

    2006-08-01

    The aim of this study was to compare an established spectrophotometrical approach for the measurement of stain removal in vitro with a new digital image analysis system. Eighteen acrylic blocks were stained by cycling them through human saliva (2 min), chlorhexidine (2 min) and tea (1 h), rinsed with deionized water and left to air dry. The absorbance of each block was then measured at 395 nm using a single-beam spectrophotometer. The lightness (L-value) of the stained blocks (after a baseline correction) was measured using digital image analysis. Image acquisition and L-values were obtained using Adobe Photoshop software. The stain removal ability of two whitening toothpastes and deionized water was tested by immersing each stained block in a test slurry (15 g paste/60 ml deionized water) for 1 min, rinsing and finally left to air dry. This cycle was repeated until the blocks had 5 min exposure to the slurry. Absorbance values from spectrophotometry and L-values by image analysis were obtained after each cycle. Fleiss' coefficient of reliability for intra-operator repeatability of the image analysis system and spectrophotometry was 0.999 for both methods which shows excellent reliability. Pearson's correlation coefficients for the two methods (stain build-up) were 0.976. Test products A, B and C gave correlations of 0.962, 0.998 and 0.817 respectively (stain removal), significant at the 0.01 level. The image system is a reliable alternative measurement method validated here against spectrophotometry for stain removal in vitro, and can provide full colour measurement.

  15. Necrotic seminoma of the testis: establishing the diagnosis with Masson trichrome stain and immunostains.

    Science.gov (United States)

    Florentine, Barbara D; Roscher, Arno A; Garrett, Jerry; Warner, Nancy E

    2002-02-01

    We describe an infarcted mass in the testis containing "ghost" cells suspicious for neoplasm. The entire lesion was necrotic. A Masson trichrome stain greatly improved nuclear and cytologic detail, confirming the suspicion of neoplasm. Placental alkaline phosphatase revealed specific membrane staining of the neoplastic cells and established a diagnosis of seminoma. Masson trichrome plus selected immunostains offer a promising approach to the diagnosis of certain necrotic neoplasms.

  16. Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle Lesions in the Head and Neck Region

    OpenAIRE

    Chang, Julia Yu Fong; Kessler, Harvey P.

    2008-01-01

    Myofibromas are well described in the head and neck region, but differentiating them from smooth muscle lesions is still difficult using smooth muscle immunohistochemical stains. This study evaluated the usefulness of the Masson trichrome stain in differentiating myofibromas from smooth muscle lesions in the head and neck region. Methods: Samples of 11 oral myofibromas, two leiomyomas, one angioleiomyoma, and one smooth muscle hamartoma were retrieved from our archives. Immunohistochemistr...

  17. Corneal staining reductions observed after treatment with Systane Lubricant Eye Drops.

    Science.gov (United States)

    Christensen, Mike T

    2008-11-01

    Because of the added emphasis on ocular surface damage included in the Dry Eye Workshop's revised definition of dry eye, an evaluation of corneal staining reductions was conducted for propylene glycol/polyethylene glycol 400-based artificial tear drops (Systane Lubricant Eye Drops; Alcon Laboratories, Fort Worth, TX, USA). An analysis was conducted on the percent change from baseline in mean corneal staining scores as reported in two previously published, randomized, double-masked, 6-week clinical studies of Systane. A descriptive comparison was also made between the outcome of the composite analysis and data obtained for Optivetrade mark Lubricant Eye Drops (Allergan, Inc., Irvine, CA, USA). Finally, results were reviewed for an open-label study that investigated corneal staining over a 5-week period after patients discontinued Systane therapy. The composite analysis included 107 Systane-treated patients. The results showed that Systane consistently reduced corneal staining at each visit; the percent change from baseline to day 42 (exit) was 47.1% (P<0.0001). After discontinuing Systane, immediate and significant increases in corneal staining were reported by 20 patients, with an overall increase from baseline to day 35 (exit) of 195.0% (P<0.0001). Evaluations of sum corneal ocular staining scores provide clinically meaningful evidence of dry eye severity, and are an important indicator of dry eye disease progression. The results of the composite analysis of two peer-reviewed studies indicate that Systane significantly reduced corneal staining (P<0.0001), indicating a reduction in the severity of dry eye. Finally, discontinuation of Systane results in a rapid increase in corneal staining that further confirms Systane's ability to maintain ocular surface health.

  18. 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.

  19. Natural fruit extracts as non-toxic fluorescent dyes for staining fungal chlamydospores.

    Science.gov (United States)

    Vujanovic, Silva; Goh, Yit Kheng; Vujanovic, Vladimir

    2012-01-01

    Currently, most synthetic dyes utilized for fungal fluorescent staining are toxic, carcinogenic, or harmful to animals, humans, and the environment. This study proposes non-toxic extracts of fruits from the genera Rhamnus, Ribes, Sambucus, Viburnum, Sorbus and Beta as simple, safe, and ecological alternatives to chemical fluorescent dye for efficient staining of Fusarium chlamydospore cells using, as test strains, five different pathogenic Fusarium species.

  20. Evaluation of Protamine Level in Human Sperm Samples Using Chromomycin A3 and Aniline Blue Staining

    Directory of Open Access Journals (Sweden)

    Durdi Qujeq

    2016-02-01

    Full Text Available Background: Current microscopic experimental methods cannot diagnose DNA damages present in spermatozoa .Therefore, some methods are needed to address the abnormality of the genetic material status on the sperm samples. As reported by many investigators aniline blue staining technique has been used for identifying sperm chromatin condensation. Also, chromomycin A3 is used for evaluation of the degree of protamination of spermatozoa. This study aimed at evaluating these two different staining techniques on human sperm protamine status. Materials and Methods: Sperm samples were collected from 72 males [including 37 infertile men: (seven asetenotratospermic, two trato-espermic, and one azo-spermic and 35 healthy fertile men]   attending the research and clinical center for infertility affiliated with Babol University of Medical Sciences. Measurement of sperm motility, volume and density of semen samples were carried out in andrology laboratory. In estimation with light microscopy aniline blue tool, in each slide, blue stained were assumed as normal spermatozoa, but dark blue stained were regarded as abnormal spermatozoa. Bright yellow stained chromomycin-reacted spermatozoa (CMA3+ were observed under fluorescent microscope with 460 nm filter considered as normal and yellowish green were assumed as abnormal. Statistical analysis results were expressed as mean ± SD. Results: The rate of reacted spermatozoa to aniline blue in the infertile group was higher than that of the healthy control group 42.8% ±8.7 vs. 17.9% ±6.4. Also, the rate of reacted spermatozoa to CMA3 in infertile and normal group was [53.6 ± 8.7 and 24.7% ±5.1], respectively. Conclusion: Infertility status could be assessed by staining the spermatozoa via aniline blue and CMA3 techniques. Combination of these two staining methods had the best predictive values for semen analysis compared to using just one method. Our results showed that both CMA3 and AB staining methods were

  1. A rapid procedure for routine double staining of cartilage and bone in fetal and adult animals.

    Science.gov (United States)

    Kimmel, C A; Trammell, C

    1981-09-01

    A simple, rapid procedure for dual staining of cartilage and bone in rodents, particularly in late gestation, has been developed for routine use. The procedure involves rapid, complete skinning of fresh eviscerated specimens following a 30 sec immersion in a 70 C water bath. The unfixed specimen is stained in a mixture of 0.14% Alcian blue and 0.12% alizarin red S in ethanol and glacial acetic acid. Specimens are then macerated in 2% KOH, cleared and hardened in 1:1 glycerin and distilled water, and stored in pure glycerin. Rapid staining of cartilage only is done in a mixture of 0.08% Alcian blue, glacial acetic acid, and ethanol, with subsequent maceration, clearing, and hardening as in the double staining procedure. Rapid staining of bone only, concurrent with maceration of soft tissue, can be done by placing fresh, unskinned specimens in a diluted mixture of alizarin red S in 2% KOH, with subsequent clearing and hardening in 1:1 distilled water and glycerin. Good quality fetal specimens can be prepared for examination by any of these procedures in a minimum of 11/2-2 days as compared to a minimum of 4-5 days for other procedures. Double stained specimens can be examined for abnormalities of the cartilage as well as bone.

  2. 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.

  3. Effect of toothpaste with natural calcium carbonate/perlite on extrinsic tooth stain.

    Science.gov (United States)

    Matheson, J R; Cox, T F; Baylor, N; Joiner, A; Patil, R; Karad, V; Ketkar, V; Bijlani, N S

    2004-01-01

    The current study was designed to determine the effect of natural calcium carbonate toothpaste containing Perlite and microgranules (Whitening toothpaste) on extrinsic tooth stain compared to a standard commercial toothpaste formulation with precipitated calcium carbonate (PCC) as abrasive and a commercial toothpaste with dicalcium phosphate dihydrate (DCPD) as abrasive. The toothpastes were evaluated in a double blind, three-cell, stratified (tobacco use; baseline tooth stain level), parallel group design study involving 600 subjects with extrinsic tooth stain. Subjects brushed twice daily with their allocated toothpaste for four weeks. Extrinsic tooth stain was measured using the Macpherson modification of the Lobene stain index. ANCOVA showed significant differences between toothpastes (p=0.037). Subsequent multiple comparisons using pairwise t-tests, showed the Whitening toothpaste to be superior to the DCPD toothpaste (p=0.014) and the PCC toothpaste (p=0.067). When a Box-Cox transformation was made to the data (y0.6) to improve normality, these two differences were more accurately estimated at p=0.004 and p=0.03 respectively. The Whitening toothpaste has been shown to be significantly more effective in tooth stain removal than the two standard commercial toothpaste formulations.

  4. Investigation Of The Color Changing Properties Of Wood Stain Derived From Pinar Leaves

    Directory of Open Access Journals (Sweden)

    Abdi Atılgan

    2011-11-01

    Full Text Available This study was designed to develop an environmentally friendly wood stain derived pinar (Quercus aucheri leaves and determine the color stability of this stain when exposed to UV light irradiation. Wood stains derived from pinar leaves were prepared from aqueous solution with %3 iron (FeSO4.7H2O , % 5 alum ((KAl(SO42.12H2O, and % 10 vinegar mordant mixtures. Scots pine (Pinus sylvestris L., Turkish oriental beech (Fagus orientalis Lipsky and oak (Quercus petraea L. wood specimens were used as staining substrates. After treatment with the stain, the wood panels were exposed to UV light irradiation for periods of 100, 200, and 300 hours and determinated the total color changes was according to ISO 2470 standards. Results showed that wood stain derived from pinar extract provided some color stability after UV irradiation. According to results, Scots pine specimens treated with the pinar extract + iron mixture provided the smallest total color changes. Meanwhile the highest total color change provided on the Scots pine treated with pinar extract+alum mixture.

  5. Staining of Platyhelminthes by herbal dyes: An eco-friendly technique for the taxonomist

    Directory of Open Access Journals (Sweden)

    Niranjan Kumar

    2015-11-01

    Full Text Available Aim: An environment compatible technique to stain Platyhelminthes, Fasciola gigantica, Gastrothylax crumenifer, Taenia solium, and Moniezia expansa using aqueous and alcoholic extract of sugar beet (Beta vulgaris, China rose (Hibiscus rosasinensis, and red rose (Rosa hybrida were described to minimized the deleterious effects of the synthetic dyes. Materials and Methods: Aqueous/ethanolic extracts of roses were extracted from the flowers while red beet was extracted from the roots. Results: Stained helminthes acquired a comparable level of pigmentation with the distinction of their internal structure in these natural dyes. The flukes (liver and rumen internal structure, oral and ventral/posterior sucker, cirrus sac, gravid uterus, testes, ovary, and vitallaria were appeared pink color in aqueous and alcoholic extract of either China or red rose and yellow to brown color in sugar beet stain. The interior of the proglottid of T. solium and M. expansa took yellow to brown color with good contrast in sugar beet stain and of pink to pink-red in China and red rose stain. Conclusion: The extract of roses (red rose followed by China rose followed by red beet possess the potential to replace the conventional stains in the taxonomic study of Platyhelminthes parasites.

  6. Statistical modeling, detection, and segmentation of stains in digitized fabric images

    Science.gov (United States)

    Gururajan, Arunkumar; Sari-Sarraf, Hamed; Hequet, Eric F.

    2007-02-01

    This paper will describe a novel and automated system based on a computer vision approach, for objective evaluation of stain release on cotton fabrics. Digitized color images of the stained fabrics are obtained, and the pixel values in the color and intensity planes of these images are probabilistically modeled as a Gaussian Mixture Model (GMM). Stain detection is posed as a decision theoretic problem, where the null hypothesis corresponds to absence of a stain. The null hypothesis and the alternate hypothesis mathematically translate into a first order GMM and a second order GMM respectively. The parameters of the GMM are estimated using a modified Expectation-Maximization (EM) algorithm. Minimum Description Length (MDL) is then used as the test statistic to decide the verity of the null hypothesis. The stain is then segmented by a decision rule based on the probability map generated by the EM algorithm. The proposed approach was tested on a dataset of 48 fabric images soiled with stains of ketchup, corn oil, mustard, ragu sauce, revlon makeup and grape juice. The decision theoretic part of the algorithm produced a correct detection rate (true positive) of 93% and a false alarm rate of 5% on these set of images.

  7. Quest for An Ideal, Simple and Cost-Effective Stain for Morphological Assessment of Sperms.

    Science.gov (United States)

    Lingappa, Hemalatha Anthanahalli; Govindashetty, Abhishek Mandya; Krishnamurthy, Anoosha; Puttaveerachary, Ashok Kagathur; Manchaiah, Sanjay; Shimoga, Indira Channagangappa; Mallaradhya, Sushma Hulikere; Gowda, Sarvesh Ballekoppa Mukunda

    2015-10-01

    Recent alarming trends of a substantial rise in the number of cases of infertility with as many as 30-40% being attributed to male-factor associated causes have created a need for further studies and advancements in semen analysis. Despite the focus on semen analysis over the years, assessment of sperm morphology has not been given due importance although it is a simple, standard and baseline diagnostic modality. It can be used to predict the need and outcome of Artificial Reproductive Techniques such as Invitro Fertilization, Gamete Intra Fallopian Tube Transfer and Intra Cytoplasmic Sperm Injection. To find the ideal, simple and cost-effective basic stain for assessment of sperm morphology in a rural tertiary care set- up where advanced equipment for assessment of sperm morphometry are inaccessible. An updated way of determining sperm shape is called the Kruger's strict morphology method. Keeping this as the standard criterion, we studied semen samples of 62 healthy male subjects using four basic staining techniques and the consensus of four independent observers was tabulated. We found that Haematoxylin and Eosin stain was the best stain for assessment of sperm head morphology. Rapid Papanicolau stain was the most ideal, simple and cost-effective stain for overall assessment of sperm morphology. Sperm morphology assessment remains the baseline necessity for the diagnosis and management of male factor associated infertility when advanced techniques are unavailable, inaccessible or unaffordable.

  8. Use of indocyanine green staining technique for phacoemulsification in white cataract

    Directory of Open Access Journals (Sweden)

    Hong-Mei Dai

    2014-08-01

    Full Text Available AIM:To observe the application efficiency of 5g/L indocyanine green(ICGstaining technique for continuous circular capsulorhexis(CCCduring phacoemulsification in white cataract.METHODS:Ninety-eight patients(98 eyeswith white cataract were randomly divided into staining group(50 cases, 50 eyesand control group(48 cases, 48 eyes. The control group didn't do anterior capsule staining. The staining group was injected to fill the anterior chamber, 5g/L ICG 0.1mL was applied on the central surface of the anterior capsule, using a 27G blunt needle through the side-port after 30s, and the redundant ICG was replaced by BSS, and continuous curvilinear capsulorhexis was accomplished using capsulorhexis forceps.RESULTS: In staining group: after ICG staining, the capsule, which presented uniform light green and visualization of the anterior capsule was significantly improved. There are 48 eyes capsulorhexis success. The rate of success was 96%. Meanwhile, in control group, there was 29 eyes capsulorhexis success. The rate of success was 60%. The difference was statistically significant(PCONCLUSION:Indocyanine green staining increases the visibility of anterior capsule in over mature cataract, and it should be an effective and helpful method which can increase the success rate of capsulorehxis. At the same time, it can reduce the incidence of intraoperative complications. This will help beginners quickly grasp continuous curvilinear capsulorhexis, and shorten the learning curve.

  9. Diagnostic performance of dual-staining cytology for cervical cancer screening: A systematic literature review.

    Science.gov (United States)

    Tjalma, Wiebren A A

    2017-03-01

    Cervical cancer screening saves lives. Secondary prevention in cervical cancer screening relies on the results of primary cytology and/or HPV testing. However, primary screening with cytology has a low sensitivity, and HPV screening has a low specificity. This means that either cancers are missed, or women are over-treated. To improve performance outcomes, the concept of dual-stain cytology (CINtec ® PLUS Cytology test) has been introduced. In this approach, additional staining with p16/Ki-67 is performed in cases where cytology results are abnormal (LSIL or ASCUS) and/or HPV-positive. Another way to describe this approach might be "diagnostic" cytology. In order to assess the value of this "diagnostic cytology", a systematic literature review was conducted of dual-stain cytology performance across multiple studies until May 2016. In a Belgian screening population (women age 25-65 years), dual-stain cytology was significantly more sensitive (66%) and slightly less specific (-1.0%) than cytology. In the population referred to colposcopy or with abnormal cytology (ASCUS, LSIL), dual-staining showed a significantly higher increase in specificity, and a slightly lower sensitivity than HPV testing. Specificity gains resulted in fewer false positives and an increase in the number of correct referrals to colposcopy. Dual-staining with p16/Ki-67 cytology is an attractive biomarker approach for triage in cervical cancer screening. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    AK Sarvel

    2006-10-01

    Full Text Available Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%. When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs; mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

  11. Staining of Platyhelminthes by herbal dyes: An eco-friendly technique for the taxonomist.

    Science.gov (United States)

    Kumar, Niranjan; Mehul, Jadav; Das, Bhupamani; Solanki, J B

    2015-11-01

    An environment compatible technique to stain Platyhelminthes, Fasciola gigantica, Gastrothylax crumenifer, Taenia solium, and Moniezia expansa using aqueous and alcoholic extract of sugar beet (Beta vulgaris), China rose (Hibiscus rosa-sinensis), and red rose (Rosa hybrida) were described to minimized the deleterious effects of the synthetic dyes. Aqueous/ethanolic extracts of roses were extracted from the flowers while red beet was extracted from the roots. Stained helminthes acquired a comparable level of pigmentation with the distinction of their internal structure in these natural dyes. The flukes (liver and rumen) internal structure, oral and ventral/posterior sucker, cirrus sac, gravid uterus, testes, ovary, and vitallaria were appeared pink color in aqueous and alcoholic extract of either China or red rose and yellow to brown color in sugar beet stain. The interior of the proglottid of T. solium and M. expansa took yellow to brown color with good contrast in sugar beet stain and of pink to pink-red in China and red rose stain. The extract of roses (red rose followed by China rose) followed by red beet possess the potential to replace the conventional stains in the taxonomic study of Platyhelminthes parasites.

  12. Randomized controlled trial to evaluate tooth stain reduction with nicotine replacement gum during a smoking cessation program

    National Research Council Canada - National Science Library

    Whelton, Helen; Kingston, Rose; O'Mullane, Denis; Nilsson, Frederick

    2012-01-01

    .... Participants were randomised to use either the Nicorette(®) Freshmint Gum or Nicorette(®) Microtab (tablet). Tooth staining and shade were rated using the modified Lobene Stain Index and the Vita...

  13. 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

  14. Comparison of modified Chicago sky blue stain and potassium hydroxide mount for the diagnosis of dermatomycoses and onychomycoses.

    Science.gov (United States)

    Liu, Zhong; Sheng, Ping; Yang, Yan-Ping; Li, Wen; Huang, Wen-Ming; Wang, Jie-Di; Fan, Yi-Ming

    2015-05-01

    The diagnostic value of modified Chicago sky blue (CSB) stain and potassium hydroxide (KOH) mount for superficial mycoses was compared using fungal culture as gold standard. The sensitivity and screening time of the CSB stain were superior to the KOH mount. The CBS stain is simple, quick and reliable for diagnosing superficial mycoses. Copyright © 2015. Published by Elsevier B.V.

  15. Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycans in sodium dodecyl sulfate-polyacrylamide gels

    DEFF Research Database (Denmark)

    Møller, H J; Heinegård, D; Poulsen, J H

    1993-01-01

    Proteoglycans stain weakly in polyacrylamide gels by traditional protein stains such as coomassie brilliant blue or silver. In the present work preparations of large aggregating proteoglycan from human articular cartilage were used to evaluate a convenient staining method based on successive stai...

  16. Effect of toothpaste containing d-limonene on natural extrinsic smoking stain: a 4-week clinical trial.

    Science.gov (United States)

    Xie, Ping; Lu, Junjun; Wan, Huchun; Hao, Yuqing

    2010-08-01

    To determine whether natural smoking stain could be removed/inhibited effectively by a toothpaste containing 5% d-limonene. For comparison and contrast, the effects of d-limonene on tea stain were also assessed. The design was a randomized controlled double-blind trial with parallel groups. Toothpastes were: A: positive control with perlite whitening formulation; B: A+5% d-limonene; C: D + 5% d-limonene; D: negative control. The extrinsic stains were measured using Lobene Stain Index. Following baseline examination, all subjects were randomly assigned to one of the four toothpaste groups and instructed to brush with the assigned products twice daily. Subjects returned to the clinic after 4-week brushing for stain removal assessment, then all extrinsic stains, plaque and supragingival calculus were removed and use of assigned products was continued for another 4 weeks, and the stain scores were repeated for inhibition assessment. A total of 408 subjects, 201 with smoking stains and 207 with tea stains, participated in the trial. 5% d-limonene combined with Perlite whitening formulation significantly reduced stain scores both for smoking stain removal and inhibition (P 0.05). The additional advantage of 5% d-limonene was shown neither for removal nor for inhibition in the tea stain study (P > 0.05). All test products were well tolerated over the study period.

  17. The use of fluorescence enhancement to improve the microscopic diagnosis of falciparum malaria

    Directory of Open Access Journals (Sweden)

    Liu Paul

    2007-07-01

    Full Text Available Abstract Background Giemsa staining of thick blood smears remains the "gold standard" for detecting malaria. However, this method is not very good for diagnosing low-level infections. A method for the simultaneous staining of Plasmodium-parasitized culture and blood smears for both bright field and fluorescence was developed and its ability to improve detection efficiency tested. Methods A total of 22 nucleic acid-specific fluorescent dyes were tested for their ability to provide easily observable staining of Plasmodium falciparum-parasitized red blood cells following Giemsa staining. Results Of the 14 dyes that demonstrated intense fluorescence staining, only SYBR Green 1, YOYO-1 and ethidum homodimer-2 could be detected using fluorescent microscopy, when cells were first stained with Giemsa. Giemsa staining was not effective when applied after the fluorescent dyes. SYBR Green 1 provided the best staining in the presence of Giemsa, as a very high percentage of the parasitized cells were simultaneously stained. When blood films were screened using fluorescence microscopy the parasites were more readily detectable due to the sharp contrast between the dark background and the specific, bright fluorescence produced by the parasites. Conclusion The dual staining method reported here allows fluorescence staining, which enhances the reader's ability to detect parasites under low parasitaemia conditions, coupled with the ability to examine the same cell under bright field conditions to detect the characteristic morphology of Plasmodium species that is observed with Giemsa staining.

  18. Effect of bleaching on staining susceptibility of resin composite restorative materials.

    Science.gov (United States)

    Celik, Ciğdem; Yüzügüllü, Bulem; Erkut, Selim; Yazici, A Rüya

    2009-01-01

    Effect of bleaching procedures on staining susceptibility of resin restorative materials is still questionable. The aim of this study was to evaluate the staining susceptibility of restorative materials bleached with 20% carbamide peroxide home bleaching agent and subsequently immersed in coffee and tea. Forty-two disk-shaped specimens were fabricated for each of the resin composites (Filtek Supreme XT [3M ESPE, St. Paul, MN, USA], Ceram-X Mono [Dentsply, Konstanz, Germany], and Aelite All Purpose Body [BISCO, Inc., Shaumburg, IL, USA]). The baseline color values were measured with a spectrophotometer. The specimens of each restorative material were randomly divided into two groups (N = 21). While the first group specimens were stored in distilled water (nonbleaching group-control), bleaching agent (Opalescence PF 20% [Ultradent Poducts, South Jordan, UT, USA]) was applied on the top surface of each specimen of the second group (bleaching group). After color change values were measured, the specimens were randomly divided into three subgroups (N = 7) according to the staining solutions. The color change values (DeltaE*ab) were calculated and the data were subjected to analysis of variance. Statistical significance was declared if the p value was 0.05 or less. There was no statistically significant difference within each restorative material's DeltaE*ab values after bleaching (p = 0.714). Also, the staining solutions did not cause a statistically significant difference between DeltaE*ab values of bleaching compared with nonbleaching groups (p = 0.146). Significant interaction was found only between restorative materials and staining solutions (p = 0.000). Bleaching of the tested resin composites did not increase their susceptibility to extrinsic staining in vitro. CLINICAL SIGNIFICANCE Bleaching did not affect staining susceptibility of the tested resin composite restorative materials. (J Esthet Restor Dent 21:407-415, 2009).

  19. A long-term laboratory test on staining susceptibility of esthetic composite resin materials.

    Science.gov (United States)

    Ardu, Stefano; Braut, Vedrana; Gutemberg, Daniel; Krejci, Ivo; Dietschi, Didier; Feilzer, Albert J

    2010-09-01

    To evaluate the color stability of composite resin types designed for esthetic anterior restorations when continuously exposed to various staining agents. Thirty-six disk-shaped specimens were made of each of 12 composite materials (1 microfilled and 11 hybrid composites). After dry storage at 37 degrees C for 24 hours in an incubator (INP-500, Memmert), the initial color of each specimen was assessed by a calibrated reflectance spectrophotometer (SpectroShade). Specimens were immersed in five staining solutions or dry stored (control). All specimens were kept in an incubator at 37 degrees C for 99 days. Test solutions were changed every 14th day to avoid bacteria or yeast contamination. After 99 days of storage, spectrophotometric measurements were again performed and L*a*b* scores once more recorded to determine the color changes. Wine proved to have the highest staining potential followed by coffee, tea, orange juice, and cola, which had the lowest staining potential. The highest color change measured against a white background was observed for Durafill (Heraeus Kulzer) in wine (DeltaE = 62.3), while the least staining was found for Enamel HFO (Micerium) in cola (DeltaE = 3.5). The highest color change measured against a black background was observed for EsthetX (Dentsply) in wine (DeltaE = 46.0), while the least staining was observed for Enamel HFO in cola (DeltaE = 2.5). Composite staining susceptibility proved to vary among composite structure and brands. Potential discoloration might be limited by dietary restriction based on such in vitro evaluation.

  20. Automatic quantification of IHC stain in breast TMA using colour analysis.

    Science.gov (United States)

    Fernández-Carrobles, M Milagro; Bueno, Gloria; García-Rojo, Marcial; González-López, Lucía; López, Carlos; Déniz, Oscar

    2017-11-01

    Immunohistochemical (IHC) biomarkers in breast tissue microarray (TMA) samples are used daily in pathology departments. In recent years, automatic methods to evaluate positive staining have been investigated since they may save time and reduce errors in the diagnosis. These errors are mostly due to subjective evaluation. The aim of this work is to develop a density tool able to automatically quantify the positive brown IHC stain in breast TMA for different biomarkers. To avoid the problem of colour variation and make a robust tool independent of the staining process, several colour standardization methods have been analysed. Four colour standardization methods have been compared against colour model segmentation. The standardization methods have been compared by means of NBS colour distance. The use of colour standardization helps to reduce noise due to stain and histological sample preparation. However, the most reliable and robust results have been obtained by combining the HSV and RGB colour models for segmentation with the HSB channels. The segmentation provides three outputs based on three saturation values for weak, medium and strong staining. Each output image can be combined according to the type of biomarker staining. The results with 12 biomarkers were evaluated and compared to the segmentation and density calculation done by expert pathologists. The Hausdorff distance, sensitivity and specificity have been used to quantitative validate the results. The tests carried out with 8000 TMA images provided an average of 95.94% accuracy applied to the total tissue cylinder area. Colour standardization was used only when the tissue core had blurring and fading stain and the expert could not evaluate them without a pre-processing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Stain removal effect of novel papain- and bromelain-containing gels applied to enamel.

    Science.gov (United States)

    Münchow, Eliseu A; Hamann, Henry J; Carvajal, M Teresa; Pinal, Rodolfo; Bottino, Marco C

    2016-11-01

    The aims of the study are to prepare novel stain removal gel-based formulations containing papain or bromelain and to investigate their stain removal effect when applied to enamel. Experimental bromelain- and papain-based stain removal gels were prepared. Next, enamel/dentin tooth samples (6 × 6 mm2, 4 mm in thickness) were obtained from bovine teeth, stained in coffee solution for 1 week, and measured with a digital spectrophotometer (Easyshade, Vita Zahnfabrik) for color assessment (baseline). The samples were then randomly allocated into four groups (n = 7), according to the stain removal agent applied: ContrastPM+ (Discus Dental, LLC), which is based on 20 wt.% carbamide peroxide (positive control); bromelain-based; papain-based; and no agent (negative control). The materials were applied once a week, three times per day, during 4 weeks, and following the directions of use from positive control. The samples were measured again with the Easyshade and using the CIEL * a * b * color system. The color change (ΔE *) results were obtained by subtracting the baseline values from the final color values obtained at each time point. The data were statistically analyzed using two-way repeated-measures analysis of variance and Student Newman Keuls's test as a post hoc test (α = 5 %). All stain removal agents produced greater color change than the negative control (p bromelain or papain) of vegetal origin may hold significant clinical potential as active agents for the preparation of stain removal agents free of hydrogen/carbamide peroxide.

  2. Iodine vapor staining for atomic number contrast in backscattered electron and X-ray imaging.

    Science.gov (United States)

    Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

    2014-12-01

    Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2 . The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. © 2014 The Authors. Microscopy Research Technique published by Wiley Periodocals, Inc.

  3. Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

    Directory of Open Access Journals (Sweden)

    Sharma Savitri

    2001-01-01

    Full Text Available Purpose: We describe a simple procedure of Immunoperoxidase (IP technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. Methods: A high titre polyclonal antibody against a local clinical isolate (axenic of Acanthamoeba species (trophozoite lysate antigen was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1 and patients showing no cysts in similar scrapings (pool-2, were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison with calcofluor white staining and culture. Results: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%. Conclusion: The immunoperoxidase technique is a simple and useful test in the diagnosis of

  4. [The application of Gallyas-Braak stainings in pathologic diagnosis of neurodegenerative diseases].

    Science.gov (United States)

    Wang, Luning; Zhu, Mingwei; Li, Xianghong; Gui, Qiuping

    2002-02-01

    To evaluate the role of Gallyas silver staining in the diagnosis of neurodegenerative diseases. Modified Gallyas-Braak staining method was used to investigate samples of the brain and spinal cord of 22 cases with neurodegenerative disease including Alzheimer's disease (AD), Parkinson's diseas (PD), Pick's disease, diffuse Lewy body disease (DLBD), progressive supranuclear palsy (PSP), diagnosed by clinical and routine pathologic method. 10 cases without clinical symptoms and pathologic abnormalities of the nervous system served as control. As compared with Bodian staining, Gallyas-Braak staining demonstrated clearly neurofibrillary tangles in the hippocampus and the cortex of frontal and temperal lobe in all the cases with Alzheimer's disease, 6 cases with dementia of other causes and 3 normal aged. However, global neurofibrillary tangles in the midbrain and the basal ganglia were found only with Gallyas-Braak staining in 4 cases with both dementia and extrapyramidal features. In addition, tuft-shaped astrocytes were shown with this method in the motor cortex, basal ganglia, midbrain of the above 4 cases and astrocytic plaques in the same area in 2 cases of the 4 cases. In this connexion, pathologic findings in 2 of the 4 cases corresponded to PSP and those of the other two cases fufiled the diagnostic criteria of corticobasal degeneration (CBD) Oligodendroglial cytoplasmic inclusions in the white matter of the brain and the spinal cord were founded in 3 of the 4 cases with multiple system atrophy (MSA). This silver staining demonstrated as well a lot of argyrophilic grains in the neuropil of the temporal lobe and the hippocampus in one case with AD. Gallyas silver staining could better reveal not only Alzheimer-like neurofibrillary tangles but also different glial inclusions in other neurodegenerative diseases such as PSP, CBD and MSA. Consequently, it is of great value in the pathologic diagnosis and study of such degenerative diseases.

  5. Safe and Effective Sarcoma Therapy through Bispecific Targeting of EGFR and uPAR.

    Science.gov (United States)

    Borgatti, Antonella; Koopmeiners, Joseph S; Sarver, Aaron L; Winter, Amber L; Stuebner, Kathleen; Todhunter, Deborah; Rizzardi, Anthony E; Henriksen, Jonathan C; Schmechel, Stephen; Forster, Colleen L; Kim, Jong-Hyuk; Froelich, Jerry; Walz, Jillian; Henson, Michael S; Breen, Matthew; Lindblad-Toh, Kerstin; Oh, Felix; Pilbeam, Kristy; Modiano, Jaime F; Vallera, Daniel A

    2017-05-01

    Sarcomas differ from carcinomas in their mesenchymal origin. Therapeutic advancements have come slowly, so alternative drugs and models are urgently needed. These studies report a new drug for sarcomas that simultaneously targets both tumor and tumor neovasculature. eBAT is a bispecific angiotoxin consisting of truncated, deimmunized Pseudomonas exotoxin fused to EGF and the amino terminal fragment of urokinase. Here, we study the drug in an in vivo "ontarget" companion dog trial as eBAT effectively kills canine hemangiosarcoma and human sarcoma cells in vitro We reasoned the model has value due to the common occurrence of spontaneous sarcomas in dogs and a limited lifespan allowing for rapid accrual and data collection. Splenectomized dogs with minimal residual disease were given one cycle of eBAT followed by adjuvant doxorubicin in an adaptive dose-finding, phase I-II study of 23 dogs with spontaneous, stage I-II, splenic hemangiosarcoma. eBAT improved 6-month survival from 450 days. eBAT abated expected toxicity associated with EGFR targeting, a finding supported by mouse studies. Urokinase plasminogen activator receptor and EGFR are targets for human sarcomas, so thorough evaluation is crucial for validation of the dog model. Thus, we validated these markers for human sarcoma targeting in the study of 212 human and 97 canine sarcoma samples. Our results support further translation of eBAT for human patients with sarcomas and perhaps other EGFR-expressing malignancies. Mol Cancer Ther; 16(5); 956-65. ©2017 AACR. ©2017 American Association for Cancer Research.

  6. Determining if DNA Stained with a Cyanine Dye Can Be Digested with Restriction Enzymes.

    Science.gov (United States)

    Maschmann, April; Masters, Cody; Davison, Melissa; Lallman, Joshua; Thompson, Drew; Kounovsky-Shafer, Kristy L

    2018-02-02

    Visualization of DNA for fluorescence microscopy utilizes a variety of dyes such as cyanine dyes. These dyes are utilized due to their high affinity and sensitivity for DNA. In order to determine if the DNA molecules are full length after the completion of the experiment, a method is required to determine if the stained molecules are full length by digesting DNA with restriction enzymes. However, stained DNA may inhibit the enzymes, so a method is needed to determine what enzymes one could use for fluorochrome stained DNA. In this method, DNA is stained with a cyanine dye overnight to allow the dye and DNA to equilibrate. Next, stained DNA is digested with a restriction enzyme, loaded into a gel and electrophoresed. The experimental DNA digest bands are compared to an in silico digest to determine the restriction enzyme activity. If there is the same number of bands as expected, then the reaction is complete. More bands than expected indicate partial digestion and less bands indicate incomplete digestion. The advantage of this method is its simplicity and it uses equipment that a scientist would need for a restriction enzyme assay and gel electrophoresis. A limitation of this method is that the enzymes available to most scientists are commercially available enzymes; however, any restriction enzymes could be used.

  7. Hematoxylin staining technique to locate sites of aluminium binding in aquatic plants

    Energy Technology Data Exchange (ETDEWEB)

    Havas, M.

    1986-10-01

    This paper describes a modified hematoxylin staining technique that can be used to locate sites of Al binding in freshwater plants and animals. This technique is fast, simple, and inexpensive to use. It is more reliable for organisms raised under controlled conditions, although it can be used on organisms isolated from the field. In the presence of Al, a purple stain appears which absorbs between 560 and 570 nm. This stain can be used as a live stain over the pH range of 3 to 9. The stain remains stable for at least 12 mo in alcohol-preserved specimens. Low concentrations of Fe (reddish-brown) and high concentrations of Pb (grey) and Cu (pink) can mask the reaction with Al (purple). Fluoride (10:1 molar ratio to Al), EDTA (1:1 ratio to Al), humic acid and PO/sub 4/ (1:10 ratio to Al) can prevent Al uptake or interfere with Al-hematein binding. Based on the technique described in this paper, sites of Al binding in aquatic plants and animals include: nucleus and cell wall of the green alga Mougeotia; cell wall of the aquatic moss Leptodictyum riparium; chloride cells, tip of the penis, and hind gut of the fairy shrimp, Branchinecta paludosa; and the anal papillae of the phantom midge, Chaoborus. 16 references.

  8. A hematoxylin staining technique to locate sites of aluminium binding in aquatic plants and animals

    Energy Technology Data Exchange (ETDEWEB)

    Havas, M.

    1986-10-01

    A modified hematoxylin staining technique that can be used to locate sites of Al binding in freshwater plants and animals is described. This technique is fast, simple, and inexpensive to use. It is more reliable for organisms raised under controlled conditions, although it can be used on organisms isolated from the field. In the presence of Al, a purple stain appears which absorbs between 560 and 570 nm. This stain can be used as a live stain over the pH range of 3 to 9. The stain remains stable for at least 12 mo in alcohol-preserved specimens. Low concentrations of Fe (reddish-brown) and high concentrations of Pb (grey) and Cu (pink) can mask the reaction with Al (purple). Fluoride (10:1 molar ratio to Al), EDTA (1:1 ratio to Al), humic acid and PO/sub 4/ (1:10 ratio to Al) can prevent Al uptake or interfere with Al-hematein binding. Based on the technique described in this paper, sites of Al binding in aquatic plants and animals include: nucleus and cell wall of the green alga Mougeotia; cell wall of the aquatic moss Leptodictyum riparium; chloride cells, tip of the penis, and hind gut of the fairy shrimp, Branchinecta paludosa; and the anal papillae of the phantom midge, Chaoborus. 16 refs.

  9. A pre-application drop containing carboxymethylcellulose can reduce multipurpose solution-induced corneal staining.

    Science.gov (United States)

    Paugh, Jerry R; Marsden, Harue J; Edrington, Timothy B; Deland, Paul N; Simmons, Peter A; Vehige, Joseph G

    2007-01-01

    Use of polyhexanide based multipurpose solutions (MPSs) for contact lens disinfection has been linked to low-grade corneal staining. In vitro data suggest that carboxymethylcellulose (CMC) may neutralize polyhexanides. The purpose of this investigation was to determine whether a pre-application drop of CMC reduces polyhexanide staining in vivo. Thirty adapted soft contact lens (SCL) wearers participated in this investigator-masked, randomized, two-way cross-over study. Subjects wore a new Group II lens (alphafilcon A, 66% water) daily for 4 weeks and disinfected lenses using a MPS containing 0.0001% polyaminopropyl biguanide. A lens lubricant containing either CMC or povidone as the primary viscolyzer was applied to the lens each day before lens wear. Biomicroscopic signs and symptomatology were assessed. The difference in scores, 0 to 4 weeks and the difference between lubricants were analyzed. The cumulative fluorescein staining scores for combined eyes demonstrated a significant increase over time (e.g., cumulative staining score; p=0.004 and ppolyhexanide MPS. This result is consistent with a proposed mechanism for CMC to neutralize cationic disinfectants and may offer clinicians another means to reduce this type of corneal staining.

  10. Effect of fabric mounting method and backing material on bloodstain patterns of drip stains on textiles.

    Science.gov (United States)

    Chang, J Y M; Michielsen, S

    2016-05-01

    Textiles may provide valuable bloodstain evidence to help piece together events or activities at violent crime scenes. However, in spite of over 75 years of research, there are still difficulties encountered in many cases in the interpretation and identification of bloodstains on textiles. In this study, we dripped porcine blood onto three types of fabric (plain woven, single jersey knit, and denim) that are supported in four different ways (hard, taut, loose, and semi-hard, i.e., fabric laid on denim). These four mounting methods represent different ways in which a textile may be present when blood from a violent act lands on it. This study investigates how the fabric mounting method and backing material affect the appearance of drip stains on textiles. We found that bloodstain patterns formed on fabric lying flat on a hard surface were very different from when the same fabric was suspended loosely. We also found that bloodstains formed on the technical back of single jersey knit were vastly different from those on the technical face. Interestingly, some drip stains showed blood passing through the textile and leaving a stain behind it that resembled insect stains. By observing, recording, and describing how a blood stained textile is found or presented at the scene, the analyst may be able to better understand bloodstains and bloodstain patterns on textiles, which could be useful to confirm or refute a witness's account of how blood came to be where it was found after a bloodshed event.

  11. 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

  12. 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.

  13. Characterisation of medieval yellow silver stained glass from Convento de Cristo in Tomar, Portugal

    Energy Technology Data Exchange (ETDEWEB)

    Delgado, J. [Dep. de Conservacao e Restauro, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Vilarigues, M. [Dep. de Conservacao e Restauro, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); VICARTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Ruivo, A. [VICARTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); REQUIMTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Corregidor, V.; Silva, R.C. da [Unidade de Fisica e Aceleradores, LFI, ITN, E.N.10, 2686-953 Sacavem (Portugal); CFNUL, Av., Prof. Gama Pinto n 2, 1649-003 Lisboa (Portugal); Alves, L.C., E-mail: lcalves@itn.pt [Unidade de Fisica e Aceleradores, LFI, ITN, E.N.10, 2686-953 Sacavem (Portugal); CFNUL, Av., Prof. Gama Pinto n 2, 1649-003 Lisboa (Portugal)

    2011-10-15

    Yellow decoration effects in stained glasses using silver staining were first applied in the beginning of the 14th century. The glass piece being decorated was usually painted on its side intended to be facing the exterior environment, and then fired to temperatures between 500 and 650 {sup o}C, resulting in colours ranging from pale lemon to deep orange. Stained glass fragments painted by this process and belonging to the Convento de Cristo, in Tomar, Portugal, were characterised using micro-PIXE, and complemented with other analytical techniques, namely UV-Vis spectroscopy and XRF. Preliminary analysis showed that a mixture of Ag and Cu was used for the production of the yellow staining. In order to understand this staining process and the influence of the firing temperature on the resulting colours, several soda and potash glasses with compositions similar to those of medieval glasses were produced and characterised. The role played by the addition of Cu in the final colours was also investigated.

  14. Lugol staining pattern in background epithelium of patients with esophageal squamous cell carcinoma.

    Science.gov (United States)

    Katagiri, Atsushi; Kaneko, Kazuhiro; Konishi, Kazuo; Ito, Hiroaki; Kushima, Miki; Mitamura, Keiji

    2004-01-01

    Squamous cell carcinoma of the esophagus often arises in the setting of chronic esophagitis. We investigated whether chronic esophagitis was associated with carcinogenesis in the esophageal squamous epithelium. Videoendoscopy with Lugol staining was performed in 70 patients with invasive carcinoma of the esophagus. We especially focused the study on background epithelium of the esophagus, then background epithelium was classified into two groups according to differences in Lugol staining patterns. Following Lugol solution spraying, background epithelium showing uniform greenish-brown staining was defined as having a uniform pattern. In contrast, when multiple Lugol-unstained speckles were present throughout the esophagus, the pattern was defined as speckled. Furthermore, we also investigated whether glycogenic acanthosis is present or not in background epithelium. Chronic esophagitis was present in 11 of 70 patients (16%) with invasive carcinoma, indicating a speckled pattern in background epithelium following Lugol solution spraying. The remaining 84% of patients with invasive carcinoma showed normally uniform Lugol staining background epithelium. Glycogenic acanthosis was found in 65 (93%) of 70 patients. Approximately 80% of patients with esophageal squamous cell carcinoma showed normal Lugol staining of background epithelium. Field carcinogenesis is postulated to be not predominant in the development of esophageal squamous cell carcinoma in our Japanese subjects. In contrast, glycogenic acanthosis of the esophagus was associated with the background epithelium accompanied with esophageal squamous cell carcinoma.

  15. Application of immunohistochemical staining to detect antigen destruction as a measure of tissue damage.

    Science.gov (United States)

    Onul, Abdullah; Colvard, Michael D; Paradise, William A; Elseth, Kim M; Vesper, Benjamin J; Gouvas, Eftychia; Deliu, Zane; Garcia, Kelly D; Pestle, William J; Radosevich, James A

    2012-09-01

    Electrocautery and directed energy devices (DEDs) such as lasers, which are used in surgery, result in tissue damage that cannot be readily detected by traditional histological methods, such as hematoxylin and eosin staining. Alternative staining methods, including 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to stain live tissue, have been reported. Despite providing superior detection of damaged tissue relative to the hematoxylin and eosin (H&E) method, the MTT method possesses a number of drawbacks, most notably that it must be carried out on live tissue samples. Herein, we report the development of a novel staining method, "antigen destruction immunohistochemistry" (ADI), which can be carried out on paraffin-embedded tissue. The ADI method takes advantage of epitope loss to define the area of tissue damage and provides many of the benefits of live tissue MTT staining without the drawbacks inherent to that method. In addition, the authors provide data to support the use of antibodies directed at a number of gene products for use in animal tissue for which there are no species-specific antibodies commercially available, as well as an example of a species-specific direct antibody. Data are provided that support the use of this method in many tissue models, as well as evidence that ADI is comparable to the live tissue MTT method.

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

    Science.gov (United States)

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

    2018-01-15

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

  17. Gram Stain

    Science.gov (United States)

    ... Transmitted Diseases Shingles Sickle Cell Anemia Sjögren Syndrome Staph Infections and MRSA Stroke Testicular Cancer Thalassemia Thyroid ... cocci is Staphylococcus aureus , the bacteria associated with staph infections . An example of gram-negative bacteria is ...

  18. Wood stains

    Science.gov (United States)

    ... Blood in the stool Burns of the food pipe (esophagus) Diarrhea Nausea Severe abdominal pain Vomiting Vomiting ... the stomach ( gastric lavage ) Washing of the skin (irrigation), perhaps every few hours for several days

  19. Gram Stain

    Science.gov (United States)

    ... is Staphylococcus aureus , the bacteria associated with staph infections . An example of gram-negative bacteria is Escherichia coli , the cause of many urinary tract infections . Fungi (in the form of yeasts or molds) ...

  20. Crossover clinical investigation of a whitening chewing gum for inhibiting dental stain formation in conjunction with tooth brushing.

    Science.gov (United States)

    Milleman, Jeffery L; Milleman, Kimberly R; Kleber, Carl J; Proskin, Howard M; Dodds, Michael; Kelley, Michael; Ramirez, Lilian

    2014-01-01

    The purpose of this clinical investigation was to evaluate the effectiveness of a marketed whitening chewing gum compared to a no-gum control in preventing the formation of extrinsic stains on the teeth of stain-forming subjects when chewed over a 12-week period of regular unsupervised use in conjunction with daily tooth brushing. This was a single-center, examiner-blind, randomized, 12-week crossover clinical trial. Stain-forming (after smoking or drinking coffee or tea) adults, starting with a stain-free baseline, either chewed the test gum (Orbit White) unsupervised four times per day, 15 minutes/chew, or used no gum along with daily brushing with a commercially available toothbrush and dentifrice for 12 weeks. At the crossover, all procedures were repeated with subjects assigned the opposite treatment. Extrinsic stain was measured at six and 12 weeks by both the Lobene Stain Index (LSI) and the Modified Lobene Stain Index (MLSI) using separate experienced examiners. After 12 weeks, LSI stain scores showed a significant 25% reduction (p = 0.0008) in new stain formation for subjects using the test chewing gum along with tooth brushing versus tooth brushing alone (no-gum control). The corresponding MLSI stain scores demonstrated a 36% reduction (p teeth. The overall findings of this clinical study demonstrated that regular use of Orbit White chewing gum, soon after smoking or drinking coffee or tea, will supplement daily tooth brushing in preventing unsightly stains from forming on the anterior teeth compared to brushing alone.

  1. High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy.

    Science.gov (United States)

    Tapia, Juan Carlos; Kasthuri, Narayanan; Hayworth, Kenneth J; Schalek, Richard; Lichtman, Jeff W; Smith, Stephen J; Buchanan, JoAnn

    2012-01-12

    Conventional heavy metal poststaining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by field emission scanning electron microscopy (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscopy (TEM) samples, our technique uses osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains, including uranyl acetate (UA), lead aspartate, copper sulfate and lead citrate, produced clean, highly contrasted TEM and scanning electron microscopy (SEM) samples of insect, fish and mammalian nervous systems. This protocol takes 7-15 d to prepare resin-embedded tissue, cut sections and produce serial section images.

  2. Efficacy of propidium iodide and FUN-1 stains for assessing viability in basidiospores of Rhizopogon roseolus.

    Science.gov (United States)

    Fernández-Miranda, Elena; Majada, Juan; Casares, Abelardo

    2017-01-01

    The use of spores in applications of ectomycorrhizal fungi requires information regarding spore viability and germination, especially in genera such as Rhizopogon with high rates of spore dormancy. The authors developed a protocol to assess spore viability of Rhizopogon roseolus using four vital stains to quantify spore viability and germination and to optimize storage procedures. They showed that propidium iodide is an excellent stain for quantifying nonviable spores. Observing red fluorescent intravacuolar structures following staining with 2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene)-1-phenylquinolinium iodide (FUN-1) can help identify viable spores that are activated. At 6 mo and 1 y, the spores kept in a water suspension survived better than those left within intact, dry gasterocarps. Our work highlights the importance of temperature, nutrients, and vitamins for maturation and germination of spores of R. roseolus during 1 y of storage.

  3. Double immunohistochemical staining with laminin 5 (γ2 chain) and collagen IV in colorectal neoplasms

    DEFF Research Database (Denmark)

    Fiehn, Anne-Marie Kanstrup; Bzorek, Michael; Warnecke, Mads

    2016-01-01

    Colorectal cancer (CRC) is one of the most common cancer diagnoses in the Western world. It is outnumbered several times by the precursor stage adenoma. The aim of this study was to describe the expression pattern with a double immunohistochemical staining for laminin 5 (γ2) and collagen IV...... in different colorectal neoplasms. This might be a supplementary tool to morphology in diagnostic dilemmas as microinvasive pT1 tumors and adenomas with pseudoinvasion. Laminin 5 has been shown to stain in invasive tumor cells, while collagen IV highlights the basement membrane (BM). Fifty-seven patients...... intact around normal crypts. In invasive tumors, laminin 5 stained intensely, and the BM was absent or focally discontinuous. The expression in adenomas and in pseudoinvasive areas was less consistent. The study suggests that double immunostaining with collagen IV and laminin 5 might be useful...

  4. Specific Staining of Wall Mannan in Yeast Cells with Fluorescein-Conjugated Concanavalin A

    Science.gov (United States)

    Tkacz, J. S.; Cybulska, E. Barbara; Lampen, J. O.

    1971-01-01

    A procedure is given for the coupling of fluorescein isothiocyanate to concanavalin A, a protein which specifically combines with a variety of polysaccharides, and for the subsequent isolation of the reactive conjugate. This fluorescent conjugate stains Saccharomyces cerevisiae but not Schizosaccharomyces pombe or Rhodotorula glutinis. The cell walls of the latter two organisms do not contain branched homopolymers of α-linked mannose. Furthermore, the staining of S. cerevisiae is competitively inhibited by either unlabeled concanavalin A or methyl-α-d-manno-pyranoside. On the basis of this evidence, it is concluded that the staining of S. cerevisiae results from the specific interaction of the fluorescein-concanavalin A conjugate with the α-mannan present in the cell wall of this yeast. Images PMID:5541005

  5. Intracellular staining and detection of cytokines by fluorescence-activated flow cytometry.

    Science.gov (United States)

    Freer, Giulia

    2014-01-01

    The detection of cytokines inside cells producing them has made a tremendous impact on the way immune reactivity is measured. Intracellular cytokine staining is the only immunological technique allowing determination of antigen-specific T cell function and phenotype at the same time; for this reason, it is one of the most popular methods to measure antigenicity in the evaluation of vaccine efficacy and in the study of infectious diseases. It is a flow cytometric technique based on staining of intracellular cytokines and cell markers (surface or cytoplasmic) with fluorescent antibodies after short term culture of stimulated immune cells in the presence of a protein secretion inhibitor, followed by fixation and permeabilization. Most experiments involve detection of five to ten different colors but many more can be detected by modern flow cytometers. Here, we discuss our experience using a standard protocol for intracellular cytokine staining.

  6. Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-Soo; Torelli, Marco; Hamers, Robert J.; Murphy, Catherine; Orr, Galya; Haynes, Christy L.

    2014-01-01

    A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. This method utilizes a commercially-available fluorescent probe to label primary amines at the surface of the cell. We demonstrate efficient staining of two bacterial strains, the Gram-negative Shewanella oneidensis MR-1 and the Gram-positive Bacillus subtilis 168. Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. We further use this method to identify localization patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.

  7. Enamel susceptibility to red wine staining after 35% hydrogen peroxide bleaching

    Directory of Open Access Journals (Sweden)

    Sandrine Bittencourt Berger

    2008-06-01

    Full Text Available Concern has been expressed regarding the staining of enamel surface by different beverages after bleaching. This study investigated the influence of 35% hydrogen peroxide bleaching agents on enamel surface stained with wine after whitening treatments. Flat and polished bovine enamel surfaces were submitted to two commercially available 35% hydrogen peroxide bleaching agents or kept in 100% humidity, as a control group (n = 10. Specimens of all groups were immersed in red wine for 48 h at 37°C, immediately, 24 h or 1 week after treatments. All specimens were ground into powder and prepared for the spectrophotometric analysis. Data were subjected to two-way analysis of variance and Fisher's PLSD test at 5% significance level. The amount of wine pigments uptake by enamel submitted to bleaching treatments was statistically higher than that of control group, independently of the evaluation time. Results suggested that wine staining susceptibility was increased by bleaching treatments.

  8. Use of two conventional staining methods to assess the acrosomal status of stallion spermatozoa.

    Science.gov (United States)

    Runcan, E E; Pozor, M A; Zambrano, G L; Benson, S; Macpherson, M L

    2014-07-01

    The acrosome is a highly specialised region of the spermatozoon that is essential for fertilisation. Defects or dysfunction of this structure have been associated with fertility problems in man and various domestic species including stallions. Current methods of evaluating the acrosome of stallion spermatozoa are time consuming and require specialised equipment, which is cost prohibitive to the average practitioner. To evaluate 2 conventional stains (Dip Quick and Spermac) and determine their usefulness in assessing acrosome integrity in stallions as compared with specific acrosomal labelling with a fluorescein-conjugated lectin - a method that has been validated for acrosome status evaluation in stallions. In vivo experimental design. Semen from 6 mature Miniature horse stallions of known fertility was collected on 5 separate occasions. To increase the number of reacted acrosomes, portions of each ejaculate were incubated with the calcium ionophore, A23187. Ejaculates were divided and semen samples were processed according to recommendations for fluorescein-conjugated peanut lectin, Pisum sativum agglutin, Dip Quick, and Spermac staining methods. Slides were evaluated independently by 2 separate investigators. Spermatozoa were classified as having intact, reacting, reacted or defective acrosomes. All parameters obtained by both investigators, using all 3 staining methods were highly correlated (P0.05) between investigators or staining method for the percentages of intact or reacted acrosomes. However, there was a significant difference between investigators and staining methods for determining reacting acrosome percentages (P<0.05). Dip Quick and Spermac stains are useful for determining intact vs. reacted acrosomes for stallion spermatozoa. © 2013 EVJ Ltd.

  9. An investigation into most effective method of treating stained teeth: an in vitro study.

    Science.gov (United States)

    Griffiths, C E; Bailey, J R; Jarad, F D; Youngson, C C

    2008-01-01

    To assess the most efficacious method of treating stained teeth: bleaching alone, veneering alone or a combination of bleaching and veneering and whether the choice alters depending on the degree of staining. Extracted teeth were sectioned to give 117 samples. These samples were split into unstained, lightly and darkly stained groups based on CIE-Lab value L*. The lightly and darkly stained groups were stained using tea. Teeth from each group were then assigned to one of four subgroups (control (C), bleaching alone (B), veneering alone (V), or a combination of bleaching and veneering (BV), each containing 13 samples. Veneering was performed using 0.8-mm thick ceramic veneer of shade B1. CIE-Lab values were recorded using a spectrophotometer and the colour difference (Delta E) was calculated for each intervention. The final colour was compared to the value for obtained from a B1 (Vita) Shade tab. Statistical significance was assessed using analysis of variance. In all three test groups, intervention resulted in a statistically significant colour change compared to the C group (pstained group, BV produced the most colour change and the value closest to B1 (p

  10. Colour stabilities of three types of orthodontic clear aligners exposed to staining agents.

    Science.gov (United States)

    Liu, Chen-Lu; Sun, Wen-Tian; Liao, Wen; Lu, Wen-Xin; Li, Qi-Wen; Jeong, Yunho; Liu, Jun; Zhao, Zhi-He

    2016-12-16

    The aim of this study was to evaluate and compare the colour stabilities of three types of orthodontic clear aligners exposed to staining agents in vitro. Sixty clear orthodontic aligners produced by three manufacturers (Invisalign, Angelalign, and Smartee) were immersed in three staining solutions (coffee, black tea, and red wine) and one control solution (distilled water). After 12-h and 7-day immersions, the aligners were washed in an ultrasonic cleaner and measured with a colourimeter. The colour changes (ΔE*) were calculated on the basis of the Commission Internationale de I'Eclairage L*a*b* colour system (CIE L*a*b*), and the results were then converted into National Bureau of Standards (NBS) units. Fourier transformation infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) were conducted to observe the molecular and morphologic alterations to the aligner surfaces, respectively. The three types of aligners exhibited slight colour changes after 12 h of staining, with the exception of the Invisalign aligners stained with coffee. The Invisalign aligners exhibited significantly higher ΔE* values (ranging from 0.30 to 27.81) than those of the Angelalign and Smartee aligners (ΔE* values ranging from 0.33 to 1.89 and 0.32 to 1.61, respectively, P<0.05). FT-IR analysis confirmed that the polymer-based structure of aligners did not exhibit significant chemical differences before and after the immersions. The SEM results revealed different surface alterations to the three types of aligner materials after the 7-day staining. The three types of aesthetic orthodontic appliances exhibited colour stability after the 12-h immersion, with the exception of the Invisalign aligners stained by coffee. The Invisalign aligners were more prone than the Angelalign and Smartee aligners to pigmentation. Aligner materials may be improved by considering aesthetic colour stability properties.

  11. TMARKER: A free software toolkit for histopathological cell counting and staining estimation

    Directory of Open Access Journals (Sweden)

    Peter J Schüffler

    2013-01-01

    Full Text Available Background: Histological tissue analysis often involves manual cell counting and staining estimation of cancerous cells. These assessments are extremely time consuming, highly subjective and prone to error, since immunohistochemically stained cancer tissues usually show high variability in cell sizes, morphological structures and staining quality. To facilitate reproducible analysis in clinical practice as well as for cancer research, objective computer assisted staining estimation is highly desirable. Methods: We employ machine learning algorithms as randomized decision trees and support vector machines for nucleus detection and classification. Superpixels as segmentation over the tissue image are classified into foreground and background and thereafter into malignant and benign, learning from the user′s feedback. As a fast alternative without nucleus classification, the existing color deconvolution method is incorporated. Results: Our program TMARKER connects already available workflows for computational pathology and immunohistochemical tissue rating with modern active learning algorithms from machine learning and computer vision. On a test dataset of human renal clear cell carcinoma and prostate carcinoma, the performance of the used algorithms is equivalent to two independent pathologists for nucleus detection and classification. Conclusion: We present a novel, free and operating system independent software package for computational cell counting and staining estimation, supporting IHC stained tissue analysis in clinic and for research. Proprietary toolboxes for similar tasks are expensive, bound to specific commercial hardware (e.g. a microscope and mostly not quantitatively validated in terms of performance and reproducibility. We are confident that the presented software package will proof valuable for the scientific community and we anticipate a broader application domain due to the possibility to interactively learn models for new

  12. Utilization of Capsules for Negative Staining of Viral Samples within Biocontainment.

    Science.gov (United States)

    Blancett, Candace D; Monninger, Mitchell K; Nguessan, Chrystal A; Kuehl, Kathleen A; Rossi, Cynthia A; Olschner, Scott P; Williams, Priscilla L; Goodman, Steven L; Sun, Mei G

    2017-07-19

    Transmission electron microscopy (TEM) is used to observe the ultrastructure of viruses and other microbial pathogens with nanometer resolution. Most biological materials do not contain dense elements capable of scattering electrons to create an image; therefore, a negative stain, which places dense heavy metal salts around the sample, is required. In order to visualize viruses in suspension under the TEM they must be applied to small grids coated with a transparent surface only nanometers thick. Due to their small size and fragility, these grids are difficult to handle and easily moved by air currents. The thin surface is easily damaged, leaving the sample difficult or impossible to image. Infectious viruses must be handled in a biosafety cabinet (BSC) and some require a biocontainment laboratory environment. Staining viruses in biosafety levels (BSL)-3 and -4 is especially challenging because these environments are more turbulent and technicians are required to wear personal protective equipment (PPE), which decreases dexterity. In this study, we evaluated a new device to assist in negative staining viruses in biocontainment. The device is a capsule that works as a specialized pipette tip. Once grids are loaded into the capsule, the user simply aspirates reagents into the capsule to deliver the virus and stains to the encapsulated grid, thus eliminating user handling of grids. Although this technique was designed specifically for use in BSL-3 or -4 biocontainment, it can ease sample preparation in any lab environment by enabling easy negative staining of virus. This same method can also be applied to prepare negative stained TEM specimens of nanoparticles, macromolecules and similar specimens.

  13. A minimally invasive procedure for esthetic achievement: enamel microabrasion of fluorosis stains.

    Science.gov (United States)

    Ramalho, Karen Muller; Eduardo, Carlos de Paula; Rocha, Rodney Garcia; Aranha, Ana Cecilia Correa

    2010-01-01

    Esthetic alterations (such as fluorosis) that result from intrinsic dental staining in enamel and dentin can be controlled or softened by noninvasive methods such as dental bleaching or enamel microabrasion. Part of the enamel is removed during microabrasion; however, this wear is clinically insignificant and does not harm the dental structure. This article presents a case in which the microabrasion technique was used to remove fluorosis staining. Based on the results of this case report, it can be concluded that this technique is efficient and can be considered a minimally invasive procedure.

  14. Sensitive phosphoprotein detection in SDS-PAGE via Anthracene Chrome Red A stain.

    Science.gov (United States)

    Hwang, Sun-Young; Choi, Jung-Kap

    2017-08-22

    Protein phosphorylation, one of the most important post-translational modifications, plays critical roles in many biological processes. Thus, it is necessary to precisely detect, identify and understand the phosphoproteins from protein mixture for the study of cell biology. We introduce a sensitive and specific detection method for phosphoproteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Anthracene Chrome Red A (ACRA) combined with the trivalent metal ion (Al3+ ) is converted to fluorescent complex and the fluorescence is sharply increased by a change of pH environment. Phosphoproteins and non-phosphoproteins can be easily distinguished by the fluorescence quenching due to the structural change of ACRA-Al3+ -phosphoprotein complex, unlike non-phosphoprotein complex. The method using ACRA is a negative staining based on the fluorescence quenching and has a high sensitivity comparable to Pro-Q Diamond stain. ACRA stain can detect 1-2 ng of α-casein and β-casein, 8-16 ng of ovalbumin (OVA) and κ-casein within 130 min. Moreover, the ACRA stain showed similar linear dynamic ranges and RSD to Pro-Q stain. The linear dynamic ranges of ACRA and the values of correlation coefficient were for OVA (8-500 ng, correlation coefficient r = 0.999), α-casein (4-500 ng, r = 0.992), β-casein (4-500 ng, r = 0.996), and κ-casein (8-500 ng, 0.998), respectively. On the other hand, the values of the relative standard deviations (RSD) ranged from 2.33 to 3.56% for ACRA. The method is sensitive, specific, simple, rapid and compatible with total protein stain such as SYPRO Ruby stain. Therefore, ACRA stain can be an advanced method for phosphoprotein detection in gels. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Validation of a Fully Automated HER2 Staining Kit in Breast Cancer

    Directory of Open Access Journals (Sweden)

    Cathy B. Moelans

    2010-01-01

    Full Text Available Background: Testing for HER2 amplification and/or overexpression is currently routine practice to guide Herceptin therapy in invasive breast cancer. At present, HER2 status is most commonly assessed by immunohistochemistry (IHC. Standardization of HER2 IHC assays is of utmost clinical and economical importance. At present, HER2 IHC is most commonly performed with the HercepTest which contains a polyclonal antibody and applies a manual staining procedure. Analytical variability in HER2 IHC testing could be diminished by a fully automatic staining system with a monoclonal antibody.

  16. Light microscopic and color television image analysis of the development of staining on chlorhexidine-treated surfaces.

    Science.gov (United States)

    Addy, M; Prayitno, S W

    1980-01-01

    Tooth staining with the use of chlorhexidine preparations is the major problem of long term application. Evidence suggests that the staining arises from a cationic/anionic interacation of chlorhexidine with components of certain dietary materials. The purpose of this in vitro study was to compare visually the development of tea and coffee staining on acrylic and tooth specimens treated with chlorhexidine and to follow the development of tea staining on perspex by light microscopy and color television image analysis. All specimens were maintained in their respective beverage for 5 days with test specimens removed three times a day and placed for 2 minutes in an 0.2% chlorhexidine solution. Both test tooth and acrylic specimens showed comparably and markedly increased staining by the beverages compared with control specimens. Color television image analysis of test specimens demonstrated more marked and rapid development of tea staining when studied on a daily basis. Microscopic examination revealed the staining to be made up of small particles of material which increased in size and coalesced with time. Again, marked differences were apparent in the stain on test and control specimens. The results of this in vitro method provided further evidence for a dietary aetiology to chlorhexidine staining and were consistent with clinical findings. Such a method may be useful to assess staining arising from the use of other anti-plaque agents.

  17. A nonlinear mapping approach to stain normalization in digital histopathology images using image-specific color deconvolution.

    Science.gov (United States)

    Khan, Adnan Mujahid; Rajpoot, Nasir; Treanor, Darren; Magee, Derek

    2014-06-01

    Histopathology diagnosis is based on visual examination of the morphology of histological sections under a microscope. With the increasing popularity of digital slide scanners, decision support systems based on the analysis of digital pathology images are in high demand. However, computerized decision support systems are fraught with problems that stem from color variations in tissue appearance due to variation in tissue preparation, variation in stain reactivity from different manufacturers/batches, user or protocol variation, and the use of scanners from different manufacturers. In this paper, we present a novel approach to stain normalization in histopathology images. The method is based on nonlinear mapping of a source image to a target image using a representation derived from color deconvolution. Color deconvolution is a method to obtain stain concentration values when the stain matrix, describing how the color is affected by the stain concentration, is given. Rather than relying on standard stain matrices, which may be inappropriate for a given image, we propose the use of a color-based classifier that incorporates a novel stain color descriptor to calculate image-specific stain matrix. In order to demonstrate the efficacy of the proposed stain matrix estimation and stain normalization methods, they are applied to the problem of tumor segmentation in breast histopathology images. The experimental results suggest that the paradigm of color normalization, as a preprocessing step, can significantly help histological image analysis algorithms to demonstrate stable performance which is insensitive to imaging conditions in general and scanner variations in particular.

  18. Purification and characterization of a surfactant-compatible lipase from Aspergillus tamarii JGIF06 exhibiting energy-efficient removal of oil stains from polycotton fabric.

    Science.gov (United States)

    Das, Arijit; Shivakumar, Srividya; Bhattacharya, Sourav; Shakya, Sujina; Swathi, S S

    2016-12-01

    An extracellular lipase with 23,666.66 U/ml/min activity was produced by Aspergillus tamarii JGIF06 under submerged fermentation in mineral salt medium containing coconut oil (2.5 % v/v), tryptone (2 % w/v) and ammonium chloride (2 % w/v), with initial pH of 5 ± 0.2, incubated at 25 °C for 7 days on a rotary shaker at 120 rpm. A 7.9-fold increase in lipase-specific activity was recorded after purification by DEAE Sepharose ion exchange and Sephadex G200 column chromatography. The apparent molecular mass of this enzyme was revealed as 50 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The optimal lipase activity was recorded at pH 4 and 37 °C. The enzyme revealed broad specificity towards different vegetable oils. The K m and V max of the lipase on olive oil was found to be 330.4 mg and 53,690 U/ml/min, respectively. The lipase activity was stable in the presence of surfactants such as cetrimonium bromide, sodium dodecyl sulphate and Tween 80, and metal ions and reagents such as Ca(2+), Ba(2+) and 2-mercaptoethanol. However, the activity was greatly reduced in the presence of organic solvents such as chloroform. The stain removal potential of the crude lipase was determined on polycotton fabric pieces stained with peanut oil. Lipase added to cold water alone significantly enhanced the removal of stain by 152 %. The addition of lipase also improved the stain removal efficiency of a commercially available detergent in the presence of either cold (25 ± 2 °C) or hot (65 ± 2 °C) water. The current findings suggest the potentiality of this enzyme for energy-efficient biocatalytic application.

  19. Differentiation-associated staining with anti-pimonidazole antibodies in head and neck tumors.

    NARCIS (Netherlands)

    Janssen, H.L.K.; Hoebers, F.J.; Sprong, D.; Goethals, L.; Williams, K.J.; Stratford, I.J.; Haustermans, K.; Balm, A.J.M.; Begg, A.C.

    2004-01-01

    BACKGROUND AND PURPOSE: Hypoxia is a strong negative prognostic factor for all three major treatment modalities for cancer. The bioreductive drug pimonidazole is currently under clinical investigation as a hypoxia marker. In human head and neck tumors, in addition to staining patterns typical of

  20. Differentiation-associated staining with anti-pimonidazole antibodies in head and neck tumors

    NARCIS (Netherlands)

    Janssen, Hilde L. K.; Hoebers, Frank J.; Sprong, Debbie; Goethals, Laurence; Williams, Kaye J.; Stratford, Ian J.; Haustermans, Karin M.; Balm, Alfons J.; Begg, Adrian C.

    2004-01-01

    BACKGROUND AND PURPOSE: Hypoxia is a strong negative prognostic factor for all three major treatment modalities for cancer. The bioreductive drug pimonidazole is currently under clinical investigation as a hypoxia marker. In human head and neck tumors, in addition to staining patterns typical of

  1. Heterogeneous staining: a tool for studies of how fluorescent dyes affect the physical properties of DNA.

    Science.gov (United States)

    Nyberg, Lena; Persson, Fredrik; Akerman, Björn; Westerlund, Fredrik

    2013-10-01

    The commonly used fluorescent dye YOYO-1 (YOYO) has, using bulk techniques, been demonstrated to stain DNA heterogeneously at substoichiometric concentrations. We here, using nanofluidic channels and fluorescence microscopy, investigate the heterogeneous staining on the single DNA molecule level and demonstrate that the dye distribution is continuous. The equilibration of YOYO on DNA is extremely slow but can be accelerated by increasing the ionic strength and/or the temperature. Furthermore, we demonstrate how to use the heterogeneous staining as a tool for detailed and time-efficient studies of how fluorescent dyes affect the physical properties of DNA. We show that the relative increase in extension of DNA with increasing amount of YOYO bound is higher at low ionic strengths and also extrapolate the extension of native DNA. Our study reveals important information on how YOYO affects the physical properties of DNA, but it also has broader applications. First, it reveals how cationic intercalators, such as potential DNA drugs, affect DNA under strong confinement. Second, the strategy of using heterogeneous staining is of general use for single molecule studies of DNA interacting with proteins or ligands.

  2. Iodamoeba butschlii in an anal pap test confirmed by iodine stain.

    Science.gov (United States)

    Schuetz, Audrey N; Pritt, Bobbi S; Schreiner, Andrew M

    2014-09-01

    We report the finding of Iodamoeba butschlii amebic cysts on a liquid-based anal Pap smear from an HIV-positive male. Iodine staining of the smear confirmed the diagnosis. It is important to distinguish I. butschlii from pathogenic ameobae and other organisms seen on anal Pap smears. Copyright © 2013 Wiley Periodicals, Inc.

  3. Visual detection of trace lead ion based on aptamer and silver staining nano-metal composite.

    Science.gov (United States)

    Ma, Li-Hong; Wang, Hai-Bo; Fang, Bi-Yun; Tan, Fang; Cao, Yuan-Cheng; Zhao, Yuan-Di

    2017-12-11

    In this paper, visual detection of trace lead ion was established by aptamer and silver staining. The basic strategy was that aminated PS2.M aptamer was immobilized onto slide and formed stable G-quadruplex structure. PbS was generated by adding S2-, and it catalyzed subsequent silver staining reaction, through the silver staining amplification effect, the slide presented visible ash black. The gray value of slide after silver staining was analyzed and the semi-quantitative detection of Pb2+ in solution was realized. The results showed that optical darkness ratio (ODR) and logarithmic value of Pb2+ concentration had a good linear relationship (R2 = 0.951) over the range of 0.5-10 μM. In addition, there was no obvious interference of other common metal ions for the detection, indicating that this method presented outstanding selectivity. And it was also used for qualitative and semi-quantitative determination of Pb2+ in soil sample successfully. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Neo-Timm staining in the thalamus of chronically epileptic rats

    Directory of Open Access Journals (Sweden)

    Hamani C.

    2005-01-01

    Full Text Available The thalamus is an important modulator of seizures and is severely affected in cholinergic models of epilepsy. In the present study, chronically epileptic rats had their brains processed for neo-Timm and acetylcholinesterase two months after the induction of status epilepticus with pilocarpine. Both controls and pilocarpine-treated animals presented neo-Timm staining in the anterodorsal nucleus, laterodorsal nucleus, reticular nucleus, most intralaminar nuclei, nucleus reuniens, and rhomboid nucleus of the thalamus, as well as in the zona incerta. The intensity of neo-Timm staining was similar in control and pilocarpine-treated rats, except for the nucleus reuniens and the rhomboid nucleus, which had a lower intensity of staining in the epileptic group. In animal models of temporal lobe epilepsy, zinc seems to modulate glutamate release and to decrease seizure activity. In this context, a reduction of neo-Timm-stained terminals in the midline thalamus could ultimately result in an increased excitatory activity, not only within its related nuclei, but also in anatomical structures that receive their efferent connections. This might contribute to the pathological substrate observed in chronic pilocarpine-treated epileptic animals.

  5. Image processing in digital pathology: an opportunity to solve inter-batch variability of immunohistochemical staining

    Science.gov (United States)

    van Eycke, Yves-Rémi; Allard, Justine; Salmon, Isabelle; Debeir, Olivier; Decaestecker, Christine

    2017-02-01

    Immunohistochemistry (IHC) is a widely used technique in pathology to evidence protein expression in tissue samples. However, this staining technique is known for presenting inter-batch variations. Whole slide imaging in digital pathology offers a possibility to overcome this problem by means of image normalisation techniques. In the present paper we propose a methodology to objectively evaluate the need of image normalisation and to identify the best way to perform it. This methodology uses tissue microarray (TMA) materials and statistical analyses to evidence the possible variations occurring at colour and intensity levels as well as to evaluate the efficiency of image normalisation methods in correcting them. We applied our methodology to test different methods of image normalisation based on blind colour deconvolution that we adapted for IHC staining. These tests were carried out for different IHC experiments on different tissue types and targeting different proteins with different subcellular localisations. Our methodology enabled us to establish and to validate inter-batch normalization transforms which correct the non-relevant IHC staining variations. The normalised image series were then processed to extract coherent quantitative features characterising the IHC staining patterns.

  6. Classification of feline intraocular neoplasms based on morphology, histochemical staining, and immunohistochemical labeling.

    Science.gov (United States)

    Grahn, Bruce H; Peiffer, Robert L; Cullen, Cheryl L; Haines, Deborah M

    2006-01-01

    The objectives of this study were to evaluate morphologic, histochemical, and immunohistochemical characteristics of well-differentiated and anaplastic intraocular neoplasms of cats, and to develop a diagnostic algorithm for, and investigate the association of ruptured lenses with these neoplasms. Seventy-five feline globes with intraocular neoplasms were stained with hematoxylin and eosin and examined by light microscopy. Morphologic diagnoses included 33 intraocular sarcomas, 17 diffuse iris melanomas, 15 lymphosarcomas, three ciliary adenomas, one metastatic carcinoma, and six undifferentiated intraocular neoplasms. Sections of these globes were then stained with periodic acid Schiff (PAS), and immunohistochemical (IHC) labels for various cellular markers. Histochemical staining and IHC labeling confirmed cellular differentiation in 73/75 neoplasms but was discordant with morphologic diagnoses in 8/75. These included four neoplasms morphologically diagnosed as lymphosarcomas but which expressed differentiation antigens consistent with melanoma (n = 3) or ciliary adenocarcinoma (n = 1), and four tumors morphologically diagnosed as intraocular sarcomas that expressed differentiation antigens for melanoma (n = 2), metastatic carcinoma (n = 1), or remained undifferentiated (n = 1). Immunohistochemical labeling suggested a diagnosis in 5/6 morphologically undifferentiated neoplasms including one intraocular sarcoma, two diffuse iridal melanomas, and two ciliary adenocarcinomas. Based upon morphologic, histochemical, and IHC characterization, ruptured lens capsules were detected in 28/30 intraocular sarcomas, 3/24 diffuse iris melanomas and 1/11 lymphosarcomas, but not in ciliary epithelial neoplasms, metastatic carcinomas, or undifferentiated intraocular neoplasms. An algorithm is provided that facilitates stain and IHC label selection for differentiating anaplastic intraocular feline neoplasms.

  7. An imidazole functionalized pentameric thiophene displays different staining patterns in normal and malignant cells

    Directory of Open Access Journals (Sweden)

    Peter eNilsson

    2015-10-01

    Full Text Available Molecular tools for fluorescent imaging of cells and their components are vital for understanding the function and activity of cells. Here, we report an imidazole functionalized pentameric oligothiophene, p-HTIm, that can be utilized for fluorescent imaging of cells. p-HTIm fluorescence in normal cells appeared in a peripheral punctate pattern partially co-localized with lysosomes, whereas a one-sided perinuclear Golgi associated localization of the dye was observed in malignant cells. The uptake of p-HTIm was temperature dependent and the intracellular target was reached within 1 h after staining. The ability of p-HTIm to stain cells was reduced when the imidazole side chain was chemically altered, verifying that specific imidazole side-chain functionalities are necessary for achieving the observed cellular staining. Our findings confirm that properly functionalized oligothiophenes can be utilized as fluorescent tools for vital staining of cells and that the selectivity towards distinct intracellular targets are highly dependent on the side-chain functionalities along the conjugated thiophene backbone.

  8. A long-term laboratory test on staining susceptibility of esthetic composite resin materials

    NARCIS (Netherlands)

    Ardu, S.; Braut, V.; Gutemberg, D.; Krejci, I.; Dietschi, D.; Feilzer, A.J.

    2010-01-01

    Objective: To evaluate the color stability of composite resin types designed for esthetic anterior restorations when continuously exposed to various staining agents. Method and Materials: Thirty-six disk-shaped specimens were made of each of 12 composite materials (1 microfilled and 11 hybrid

  9. Toothpastes containing abrasive and chemical whitening agents: efficacy in reducing extrinsic dental staining.

    Science.gov (United States)

    Soares, Cristina Neves Girao Salgado; Amaral, Flavia Lucisano Botelho do; Mesquita, Marcelo Ferraz; Franca, Fabiana Mantovani Gomes; Basting, Roberta Tarkany; Turssi, Cecilia Pedroso

    2015-01-01

    This in vitro study evaluated the efficacy of toothpastes containing abrasive and chemical whitening agents in reducing the extrinsic discoloration of dental enamel. Sixty slabs of dentin from human teeth were sealed so that only the enamel surface was exposed. The enamel surfaces were photographed for initial color assessment. Staining was performed by immersing the dental slabs in 0.2% chlorhexidine solution for 2 minutes and then in black tea for 60 minutes. This process was repeated 15 times. Photographs were taken at the end of the staining process, and the slabs were divided into 5 groups (n = 12), 3 to be brushed with toothpastes containing chemical whitening agents (2 containing phosphate salts and 1 containing phosphate salts plus hydrogen peroxide) and 2 to represent control groups (ordinary/nonwhitening toothpaste and distilled water). The dental slabs were subjected to mechanical toothbrushing with toothpaste slurry or distilled water, according to each group's specifications. After brushing, more photographs were taken for color analysis. The results showed a significant reduction in luminosity after the staining process in addition to an increase in the colors red and yellow (P whitening toothpastes and the changes found in slabs brushed with ordinary toothpaste. The whitening toothpastes did not outperform an ordinary toothpaste in the removal of extrinsic staining.

  10. Modeling laser treatment of port wine stains with a computer-reconstructed biopsy

    NARCIS (Netherlands)

    Pfefer, T. J.; Barton, J. K.; Smithies, D. J.; Milner, T. E.; Nelson, J. S.; van Gemert, M. J.; Welch, A. J.

    1999-01-01

    The efficacy of laser treatment of port wine stains (PWS) has been shown to be highly dependent on patient-specific vasculature. The effect of tissue structure on optical and thermal mechanisms was investigated for different pulse durations by using a novel theoretical model that incorporates tissue

  11. Wood staining fungi revealed taxonomic novelties in Pezizomycotina : New order Superstratomycetales and new species Cyanodermella oleoligni

    NARCIS (Netherlands)

    van Nieuwenhuijzen, E. J.; Miadlikowska, J M; Houbraken, J. A. M. P.; Adan, Olaf C G; Lutzoni, F M; Samson, R. A.

    2016-01-01

    A culture-based survey of staining fungi on oil-treated timber after outdoor exposure in Australia and the Netherlands uncovered new taxa in Pezizomycotina. Their taxonomic novelty was confirmed by phylogenetic analyses of multi-locus sequences (ITS, nrSSU, nrLSU, mitSSU, RPB1, RPB2, and EF-1α)

  12. A Technique for Staining Extracted Teeth: A Research and Teaching Aid for Bleaching.

    Science.gov (United States)

    1980-10-01

    M4yerson Standard Shade Guide, Myerson Tooth Corp., Cambridge, MA 2 before and after the staining procedure. Lingual access openings were prepared in the...teeth, the pulps extirpated, and the root canals instrumented with #10 to #20 files. The teeth were then placed in 5.25% sodium hypochlorite solution for

  13. Electron microscopy of helical filaments: rediscovering buried treasures in negative stain.

    Science.gov (United States)

    Egelman, Edward H; Amos, Linda A

    2009-09-01

    Although negative stain electron microscopy is a wonderfully simple way of directly visualizing protein complexes and other biological macromolecules, the images are not really comparable to those of objects seen in everyday life. The failure to appreciate this has recently led to an incorrect interpretation of RecA-family filament structures.

  14. Nile Red Staining for Oil Determination in Microalgal Cells: A New Insight through Statistical Modelling

    Directory of Open Access Journals (Sweden)

    Ronald Halim

    2015-01-01

    Full Text Available In the wake of global warming and rapid fossil fuel depletion, microalgae emerge as promising feedstocks for sustainable biofuel production. Nile red staining acts as a rapid diagnostic tool to measure the amount of biodiesel-convertible lipid that the cells accumulate. There is a need for the development of a more uniform staining procedure. In its first phase, this study examined the dependence of microalgal Nile red fluorescence (Tetraselmis suecica in terms of its most pertinent staining variables. A quadratic surface model that successfully described the Nile red fluorescence intensity as a composite function of its variables was generated (r2=0.86. Cell concentration was shown to have a significant effect on the fluorescence intensity. Up to a certain threshold, fluorescence intensity was shown to increase with Nile red dye concentration. In its second phase, the study reviewed findings from previous Nile red studies to elucidate some of the fundamental mechanism underlying the diffusion of Nile red dye molecules into the microalgal cells and their subsequent interaction with intracellular lipids. Through the review process, we were able to develop a simple framework that provided a set of guidelines for the standardization of the Nile red staining procedure across different microalgal species.

  15. Staining of cerebellar cortex granular layer interneurons with natural dye of Madder.

    Science.gov (United States)

    Gharravi, Anneh Mohammad

    2016-01-01

    The objective of the present study was an investigation of root Rubia Tinctorum (Madder) as a natural dye to identification of granular layer interneurons of the rat cerebellum. Seven to ten micrometre sections were collected from the cerebellum and stained only with Madder for 2, 24 and 48 h. Other sections were stained with Madder then with hematoxyllin, cresyl violet, eosin, light green. Microscopic identification of cells was performed based on cell morphology, reaction and binding of with the dye. All data were expressed as mean ± SD in and significance was set at p ≤0.05. Madder with alum as mordant resulted a deep red staining of interneurons. Unipolar brush cells (UBCs) were observed with a cell body diameter intermediate between granule and Golgi cells in the superficial layer of the granular layer. Golgi cells were identified almost as large as Purkinje cells with irregular rounded or polygonal morphology. Lugaro cells were observed as spindle-shaped cells adjacent to Purkinje layer. Results of the present study showed that mader could stain granular layer interneurons in cerebellum cortex of rat.

  16. Image processing in digital pathology: an opportunity to solve inter-batch variability of immunohistochemical staining.

    Science.gov (United States)

    Van Eycke, Yves-Rémi; Allard, Justine; Salmon, Isabelle; Debeir, Olivier; Decaestecker, Christine

    2017-02-21

    Immunohistochemistry (IHC) is a widely used technique in pathology to evidence protein expression in tissue samples. However, this staining technique is known for presenting inter-batch variations. Whole slide imaging in digital pathology offers a possibility to overcome this problem by means of image normalisation techniques. In the present paper we propose a methodology to objectively evaluate the need of image normalisation and to identify the best way to perform it. This methodology uses tissue microarray (TMA) materials and statistical analyses to evidence the possible variations occurring at colour and intensity levels as well as to evaluate the efficiency of image normalisation methods in correcting them. We applied our methodology to test different methods of image normalisation based on blind colour deconvolution that we adapted for IHC staining. These tests were carried out for different IHC experiments on different tissue types and targeting different proteins with different subcellular localisations. Our methodology enabled us to establish and to validate inter-batch normalization transforms which correct the non-relevant IHC staining variations. The normalised image series were then processed to extract coherent quantitative features characterising the IHC staining patterns.

  17. 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.

  18. Gene amplification as double minutes or homogeneously staining regions in solid tumors : Origin and structure

    NARCIS (Netherlands)

    Storlazzi, Clelia Tiziana; Lonoce, Angelo; Guastadisegni, Maria C.; Trombetta, Domenico; D'Addabbo, Pietro; Daniele, Giulia; L'Abbate, Alberto; Macchia, Gemma; Surace, Cecilia; Kok, Klaas; Ullmann, Reinhard; Purgato, Stefania; Palumbo, Orazio; Carella, Massimo; Ambros, Peter F.; Rocchi, Mariano

    Double minutes (dmin) and homogeneously staining regions (hsr) are the cytogenetic hallmarks of genomic amplification in cancer. Different mechanisms have been proposed to explain their genesis. Recently, our group showed that the MY-containing dmin in leukemia cases arise by excision and

  19. 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.

  20. 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