Mucin Histochemistry in Tumours of Colon, Ovaries and Lung
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A Valuable Stain for Connective Tissue, Keratin and Fungi* Michel Prunieras, M.D
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector PAB: A VALUABLE STAIN FOR CONNECTIVE TISSUE, KERATIN AND FUNGI* MICHEL PRUNIERAS, M.D. Since the papers by Steedman (1), Lison (2)tion. Most of the blocks were freshly prepared and Mowry (3), the use of Alcian Blue stain hasbut some were as old as 30 years. undergone considerable change. The PAB stain (routine) runs as follow: As pointed out by Pearse (4), staining with Alcian Blue is increased when acidic groups are Deparafflo and bring sections to water. introduced by sulfation or by oxidation, due Oxidize in Permanganate for 10 minutes (2.5% to the salt linkage of the dye with acidic groups.MnO4K: 100 cc.; 5% S04112: 100 cc.; distilled water: 700 cc.). The specificity of the stain might also be im- Bleach in 2% oxalic acid, 30 seconds. proved by lowering the pH of the staining bath, Wash in running water and rinse in distilled thus making Alcian Blue staining specific forwater. strong acidic groups, as shown by Adams and Stain ooe slide 30 minutes in 0.1% Alcian Blue Sloper (5). Different oxidative procedures,8GX (Imperial Chemical Industries) in 3% acetic followed by Alcian Blue stain at various pHacid (pH 2.7, 3.0) and one other slide 10 minutes have been already described in the literature:in 1% Alcian Blue in 10% sulphuric acid (pH 0.2, 0.4). A third slide might be stained 1 minute in performie acid (Adams and Sloper, 5), per-1% Alcian Blue in distilled water. -
JB-4 Kit AGR1130
Unit 7, M11 Business Link Parsonage Lane, Stansted Essex, UK CM24 8GF t: +44 (0)1279 813519 f: +44 (0)1279 815106 e: [email protected] w: www.agarscientific.com JB-4 Kit AGR1130 Introduction: JB-4 Embedding Kit is a unique polymer embedding material that gives a higher level of morphological detail than paraffin processed tissues. A water-soluble media, JB-4 does not require dehydration to absolute alcohol except for dense, bloody, or fatty tissue specimens. JB-4 is excellent for non-decalcified bone specimens, routine stains, special stains, and histochemical staining. Clearing agents such as xylene and chloroform are not required. The polymerization of JB-4 is exothermic, which is easily controlled by polymerizing on ice or by using refrigeration at 4°C. JB-4 Embedding Kits must be used under a chemical fume hood. Sections of JB-4 embedded material can be cut at 0.5 to 3.0 microns or thicker. Microtomes designed for plastic sectioning are required as are glass, Ralph, or tungsten carbide knives. Polysciences, Inc. has tungsten carbide knives available for most sectioning requirements. Sections can be stained for routine histological or histochemical procedures. Immunohistochemical procedures are not recommended for JB-4 as the glycol methacrylate cannot be removed from the section and may block antigen sites for most antibody reactions. As an alternative we recommend the Polysciences, Inc. Osteo-Bed Bone Embedding Kit. The Osteo-Bed formulation is a methyl methacrylate that is well suited for bone or for immunohistochemistry on routine histological specimens. NOTE: It is recommended that the Embedding Kit be used under a fume hood with appropriate gloves. -
I. the Preparation and Morphology of a Quantified Urine Sediment
Upsala J Med Sci 84: 67-74, 1979 The Effect of Short-term High-dose Treatment with Methenamine Hippurate of Urinary Infection in Geriatric Patients with Indwelling Catheters I. The preparation and morphology of a quantified urine sediment Bo Norberg, Astrid Norberg, Ulf Parkhede, Hans Gippert and MBns Akerman Departments of Internal Medicine, Pathology and Education, Univer.tity of Lund and the School of Nursing, Lund, Sweden. 3 A4 Riker Laboratories, Skurholmen, Swyden ABSTRACT A quantified sediment of the urine from patients with indwelling catheters was prepared by fixation of 0.1 ml urine in 0.9 ml 2% glutaraldehyde immediately after sampling. Slide preparations were then made from 0.2 ml of the glutaral- dehyde suspension by means of a cytocentrifuge. Bacteria and epithelial cells were properly contrasted by the May-Grunwald-Giemsa stain but haematoxylin- eosin and the Papanicolaou stain were superior as regards leukocyte morphology. It is suggested that glutaraldehyde-cytocentrifuge preparations of the urine cytology may be useful in the evaluation of urinary infection and in the evaluation of the therapy of urinary infection. INTRODUCTION The microscopic examination of urine sediments is a rapid and simple proce- dure which provides essential information in many cases of kidney disease or infections in the urinary tract. The conventional urinary sediment has, how- ever, serious pitfalls, e.g. low reproducibility, low precision and high vul- nerability to delay in transport and preparation (1-10). It nevertheless seemed desirable to make a quantified urine sediment from patients with indwelling catheters in order to evaluate urinary infection and the effects of therapy. -
T-Cell Brain Infiltration and Immature Antigen-Presenting Cells in Transgenic Models of Alzheimerв€™S Disease-Like Cerebral
Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2016 T-cell brain infiltration and immature antigen-presenting cells in transgenic models of Alzheimer’s disease-like cerebral amyloidosis Ferretti, M T ; Merlini, M ; Späni, C ; Gericke, C ; Schweizer, N ; Enzmann, G ; Engelhardt, B ; Kulic, L ; Suter, T ; Nitsch, R M Abstract: Cerebral beta-amyloidosis, one of the pathological hallmarks of Alzheimer’s disease (AD), elicits a well-characterised, microglia-mediated local innate immune response. In contrast, it is not clear whether cells of the adaptive immune system, in particular T-cells, react to cerebral amyloidosis in AD. Even though parenchymal T-cells have been described in post-mortem brains of AD patients, it is not known whether infiltrating T-cells are specifically recruited to the extracellular deposits of beta-amyloid, and whether they are locally activated into proliferating, effector cells upon interaction with antigen- presenting cells (APCs). To address these issues we have analysed by confocal microscopy and flow- cytometry the localisation and activation status of both T-cells and APCs in transgenic (tg) mice models of AD-like cerebral amyloidosis. Increased numbers of infiltrating T-cells were found in amyloid-burdened brain regions of tg mice, with concomitant up-regulation of endothelial adhesion molecules ICAM-1 and VCAM-1, compared to non-tg littermates. The infiltrating T-cells in tg brains did not co-localise with amyloid plaques, produced less interferon-gamma than those in controls and did not proliferate locally. Bona-fide dendritic cells were virtually absent from the brain parenchyma of both non-tg andtgmice, and APCs from tg brains showed an immature phenotype, with accumulation of MHC-II in intracellular compartments. -
Simple Technique to Identify Haemosiderin in Immunoperoxidase Stained Sections
J Clin Pathol: first published as 10.1136/jcp.37.10.1190 on 1 October 1984. Downloaded from 1190 Technical methods Phosphate buffer at pH 8*0 gave the sharpest 2 Rozenszajn L, Leibovich M, Shoham D, Epstein J. The esterase staining reactions, although there was little differ- activity in megaloblasts, leukaemic and normal haemopoietic cells. Br J Haematol 1968; 14:605-19. ence at pH 7-0 or pH 7-5. As the buffer pH was 3Hayhoe FGJ, Quaglino D. Haematological cytochemistry. Edin- increased above pH 8-0 staining with both substrates burgh: Churchill Livingstone, 1980. became progressively weaker, especially above pH 4Li CY, Lam KW, Yam LT. Esterases in human leucocytes. J 9.0. Below pH 7-0 staining with a-naphthyl butyrate Histochem Cytochem 1973;21:1-12. Yam LT, Li CY, Crosby WH. Cytochemical identification of became weaker, and below pH 5*0 staining with monocytes and granulocytes. Am J Clin Pathol 1971;55:283- naphthol AS-D chloroacetate began to disappear. 90. 6 Armitage RJ, Linch DC, Worman CP, Cawley JC. The morphol- This work was supported by a Medical Research ogy and cytochemistry of human T-cell subpopulations defined by monoclonal antibodies and Fc receptors. Br J Haematol Council project grant. I thank Professor FGJ 1983;51:605-13. Hayhoe for valuable advice. References Requests for reprints to: Dr DM Swirsky, Department of Gomori G. Chloroacyl esters as histochemical substrates. J His- Haematological Medicine, University Clinical School, Hills tochem Cytochem 1953;1:469-70. Road, Cambridge CB2 2QL, England. Simple technique to identify identification of the two compounds on the same haemosiderin in slide. -
New Tetrachromic VOF Stain (Type III-G.S) for Normal and Pathological Fish Tissues C
ORIGINAL PAPER New Tetrachromic VOF Stain (Type III-G.S) for Normal and Pathological Fish Tissues C. Sarasquete,* M. Gutiérrez Instituto de Ciencias Marinas de Andalucía, CSIC Polígono Río San Pedro, Apdo oficial, Puerto Real, Cádiz, Spain richrome methods invariably use dyes in acid ©2005, European Journal of Histochemistry pH solvents, usually diluted in aqueous acetic Tacid, and the concentration of this acid A new VOF Type III-G.S stain was applied to histological sec- matches the concentration of dye. Staining depends tions of different organs and tissues of healthy and pathologi- largely on the attachment of dyes to proteins. The cal larvae, juvenile and adult fish species (Solea senegalensis; acid pH itself is necessary to maximise the amount Sparus aurata; Diplodus sargo; Pagrus auriga; Argyrosomus regius and Halobatrachus didactylus). In comparison to the of dye that will attach to tissue amino groups. original Gutiérrez´VOF stain, more acid dyes of contrasting Proteins have both positively (amino groups) and colours and polychromatic/metachromatic properties were negatively (carboxyl and hydroxyl) charged groups. incorporated as essential constituents of the tetrachromic VOF Usually one predominates and this will have an stain. This facilitates the selective staining of different basic tissues and improves the morphological analysis of histo- overall negative or positive charge (being an acid or chemical approaches of the cell components. The VOF-Type III a basic protein). These charges can, however, bal- G.S stain is composed of a mixture of several dyes of varying ance each other out to some degree. Phosphate size and molecular weight (Orange G< acid Fuchsin< Light green<Methyl Blue<Fast Green), which are used simultane- groups of DNA and binding-proteins are important ously, and it enables the individual tissues to be selectively dif- in nuclear staining.The ionisation of basic groups of ferentiated and stained. -
Hito Oil Red O Optimstain™ Kit Manual and MSDS
Simple Solution for Your Research Hito Oil Red O OptimStain™ Kit [Catalog Number: HTKLS0122] An easy to use Oil Red O staining system for the lipid and fat staining on frozen sections User Manual And Material Safety Data Sheet FOR IN VITRO RESEARCH USE ONLY Hitobiotec Corp. Simple solution for your research Hito Oil Red O OptimStain™ Kit [Catalog Number: HTKLS0122] An easy to use Oil Red O staining system for the lipid and fat staining on frozen sections User Manual And Material Safety Data Sheet FOR IN VITRO RESEARCH USE ONLY Hitobiotec Corp. © 2017 All Rights Reserved Index I. Introduction 2 II. Kit Contents 3 III. Tissue Preparation 4 IV. Staining Procedure 8 V. References 11 VI. Material Safety Data Sheet (MSDS) 12 1 I. Introduction Hito Oil Red O OptimStain™ Kit is designed based on the Oil Red O staining method. Oil Red O is a common staining technique for studying the morphology of lipids in adipose tissue. It allows localization and visualization of the lipids in the aortic sinus and aorta to determine the extent of atherosclerosis, and remains as one of the primary techniques for visualization of the atherosclerosis pattern at the aortic sinus. Accordingly, Oil Red O staining techniques are not only useful for tissue histology studies, but also widely used in cell biological studies examining intracellular lipid metabolism 1-5. Hito Oil Red O OptimStain™ Kit makes further improve- ment over the Oil Red O method and simplifies the staining technique. It offers high quality, rapid, reliable and easy to use staining of the lipids and fat. -
Preclinical Studies of the First in Human Sarna Drug Candidate for Liver Cancer
Oncogene (2018) 37:3216–3228 https://doi.org/10.1038/s41388-018-0126-2 ARTICLE Gene activation of CEBPA using saRNA: preclinical studies of the first in human saRNA drug candidate for liver cancer 1 2 2 3 4 1 Vikash Reebye ● Kai-Wen Huang ● Vivian Lin ● Sheba Jarvis ● Pedro Cutilas ● Stephanie Dorman ● 5 1 5 6,7 1 1 Simona Ciriello ● Pinelopi Andrikakou ● Jon Voutila ● Pal Saetrom ● Paul J. Mintz ● Isabella Reccia ● 8 9 5 10 5 1 John J. Rossi ● Hans Huber ● Robert Habib ● Nikos Kostomitsopoulos ● David C. Blakey ● Nagy A. Habib Received: 2 July 2017 / Revised: 2 December 2017 / Accepted: 12 December 2017 / Published online: 7 March 2018 © The Author(s) 2018. This article is published with open access Abstract Liver diseases are a growing epidemic worldwide. If unresolved, liver fibrosis develops and can lead to cirrhosis and clinical decompensation. Around 5% of cirrhotic liver diseased patients develop hepatocellular carcinoma (HCC), which in its advanced stages has limited therapeutic options and negative survival outcomes. CEPBA is a master regulator of hepatic function where its expression is known to be suppressed in many forms of liver disease including HCC. Injection of MTL-CEBPA, a small activating RNA oligonucleotide therapy (CEBPA-51) formulated in liposomal nanoparticles 1234567890();,: 1234567890();,: (NOV340- SMARTICLES) upregulates hepatic CEBPA expression. Here we show how MTL-CEBPA therapy promotes disease reversal in rodent models of cirrhosis, fibrosis, hepatosteatosis, and significantly reduces tumor burden in cirrhotic HCC. Restoration of liver function markers were observed in a carbon-tetrachloride-induced rat model of fibrosis following 2 weeks of MTL-CEBPA therapy. -
Newsletter 4
ANATECH LTD INNOVATOR Special Stains Issue Special Stains Issue Hematoxylin and eosin (H & E) is the gold stan- dard for demonstration of tissue structure in anatomic pathology. However, by utilizing vari- ous dye solutions, special stains allow further visualization of major macromolecules (e.g., carbohydrates, proteins, minerals) in a rainbow of colors beyond the blue and pink hues of H & E staining. This makes special stains indispensable in the demonstration of tissue morphology and its components. While immunohistochemistry and molecular biology are truly advancing in 1 A B the diagnosis of diseases, the comparatively low cost of histochemical special stains makes them April 2010 vital to the pathology laboratory. Figure 1. Fatty liver metamorphosis. A) Iron, 20x; B) H&E, 40x. ANATECH LTD. has a growing family of really A new look special stains. We refer to them as really special because several of them are unique and were at some old favorites developed in response to a problem with the existing traditional stain, due to unavailability Iron or technical performance. By understanding the chemistry of dyes, ANATECH LTD. was able to Hemosiderin, an iron-storage complex, is normally respond to these problems and produce special present intracellularly in macrophages. However, stains that are chemically unique and/or offer during hemorrhaging, when red blood cells (RBC) are a technical improvement over the conventional released from the circulatory system, excess hemosid- stain. Knowing that the stained tissue’s appear- erin deposits will occur in the surrounding extracel- ance is critical, our really special stains yield lular spaces. This is seen grossly in the color change of similarly colored results as the traditional dyes. -
Living up to Life Special Stain Kit Alcian Blue
Living up to Life Special Stain Kit Alcian Blue Stain Kit Catalog No: 38016SS3 Intended Use Staining Protocol (Microwave) For In Vitro Diagnostic Use. For Laboratory Use. Exercise caution when using the microwave to heat any solution or reagent. The microwave The reagents in this kit are intended for In Vitro use only. Alcian Blue, when used with the must be properly ventilated to prevent the accumulation of fumes in the laboratory. Microwave appropriate histological staining protocol, may be useful for the demonstration of acidic mucins transparent coplin jars and caps should be used during the staining process. The caps should in tissue sections. The pH 2.5 Alcian Blue provided in this kit reacts with both carboxylated and be loosely applied to prevent spills. Caps with ventilation holes also may be used. sulfated mucins. All microwaves should be used in accordance with the manufacturer’s instructions. The procedures described here were performed using an Energy Beam Sciences H2250 laboratory Probable Mode of Action microwave. All microwaved steps were conducted at a power setting of 800 watts unless The Alcian Blue molecule is a large conjugated dye molecule that contains a central copper otherwise noted. Because of differences in microwave power and frequencies among various containing phthalocyanine ring and four isothiouronium groups. The isothiouronium groups models, it may be necessary to adjust power levels or times to achieve optimal results. are basic and impart an overall positive charge to the Alcian Blue molecule. The cationic 1. Deparaffinize with xylene or a xylene substitute and rehydrate though graded alcohols to isothiouronium groups likely bind via electrostatic interactions to the anionic sulfate and deionized water.a carboxylate groups located within the carbohydrate moieties of mucin. -
Orcein-Alcian Blue Staining: a New Technique for Demonstrating Acid Mucins in Gastrointestinal Epithelium
J Clin Pathol: first published as 10.1136/jcp.42.8.881 on 1 August 1989. Downloaded from J Clin Pathol 1989;42:881-884 Orcein-alcian blue staining: a new technique for demonstrating acid mucins in gastrointestinal epithelium R SINGH, A W P GORTON Department ofHistopathology, Manor Hospital, Walsall, West Midlands SUMMARY Orcein-alcian blue staining, a new method for the simultaneous demonstration of sulphated and sialomucins in gastrointestinal epithelium was compared with the standard high iron diamine-alcian blue technique. Sections were oxidised with potassium permanagate and decolourised in oxalic acid. They were stained with orcein for four hours, differentiated for a few seconds in acid alcohol, and then counterstained with alcian blue for halfto one minute. There was a good correlation of results between the two methods. Orcein-alcian blue is a safer, cheaper, and quicker method than high iron diamine-alcian blue which can be safely introduced into routine laboratories for the study of acid mucins in the gastrointestinal diseases. Orcein, a naturally occurring vegetable dye, has been Table used for staining elastic fibres for many years. It is also used for the demonstration of hepatitis B antigen and Histological diagnosis No ofcases copper-associated proteins in chronic liver diseases.'2 copyright. Recently, the orcein technique has been studied for Normal mucosa (stomach, ileum, appendix, large bowel) 25 Intestinal metaplasia of stomach 5 showing the presence of sulphated mucin in gastroin- Adenomas (stomach, appendix, large bowel) 6 testinal epithelium.34 It has also been reported to be Adenocarcinomas (stomach, appendix, ileum, large bowel) II selectively positive for mucin-producing adenocarcin- Total 47 omas of the lower gastrointestinal tract; therefore, adenocarcinomas in sites other than the colon and rectum are negative for orcein staining.5 We report our BDH Poole, Dorset) in 100 ml 70% alcohol and then http://jcp.bmj.com/ findings of combining a modified orcein staining adding 1-0 ml concentrated hydrochloric acid. -
Techniques for Study of Avian Syringes
SHORT COMMUNICATIONS 289 R. Banks, J. Becker, P. Cannell, F. Gill, S. Lanyon, S. Olson, K. Parkes and J. Pitocchelli reviewed the manuscript. LITERATURE CITED AMERICAN ORNITHOLOGISTS ’ UNION. 1983. Check-list of North American birds, 6th ed. Amer. Ornithol. Union, Washington, D.C. AVISE, J. C., J. C. PATTON, AND C. F. AQUADRO. 1980. Evolutionary genetics of birds. Comparative molecular evolution in New World warblers and rodents. J. Heredity 7 1: 303-3 10. BALDWIN, S. P., H. C. OBERHOLSER,AND L. G. WORLEY. 1931. Measurements of birds. Cleveland Mus. Nat. Hist. Vol. 2. BREWSTER,W. 1874. A new species of North American warbler. Am. Sportsman 5:33. -_ 188 1. On the relationship of Hebninthophaga leucobronchialis,Brewster, and HelminthophagaLawrencei, Henick, with some conjectures respecting certain other North American birds. Bull. Nuttall Ornith. Club 6:2 18-225. GILL, F. B. 1980. Historical aspects of hybridization between Blue-winged and Golden- winged warblers. Auk 97: l-l 8. HERRICK, H. 1875. Description of a new species of Helminthophaga. Proc. Acad. Sci. Philadelphia 26 (1874):220. LANGDON, F. W. 1880. Description of a new warbler of the genus Helminthophaga. J. Cincinnati Sot. Nat. Hist. 3: 119-120. MCCAMEY, F. 1950. A puzzling hybrid warbler from Michigan. Jack Pine Warbler 28: 67-72. PARKES,K. C. 195 1. The genetics of the Golden-winged x Blue-winged Warbler complex. Wilson Bull. 63:5-l 5. -. 1978. Still another parulid intergeneric hybrid (Mniotilta x Dendroica) and its taxonomic and evolutionary implications. Auk 95:682-690. RIDGWAY, R. 1880. Note on Helminthophagacincinnatiensis, Langdon. Bull.