A Simple Technique for Staining of Cell Membranes with

A Simple Technique for Staining of Cell Membranes with -...

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0022-1554195/$3.30 The Journal of Histochemistry and Cytochemistry Copyright 0 1995 by The Histochemical Society, Inc Technical Note I Vol. 43. No. 10. pp. 1079-1084. 1995 Printed in US.A. A Simple Technique for Staining Cell Membranes with Imidazole and Osmium Tetroxide' GEORGES THIERY, JACQUES BERNIER, and MICHEL BERGERON2 Deparimen i PbysioLogy, Universiii de MoniriaL, Montriad Qu6&ec, Canada. Received for publication February 8, 1995 and in revised form May 3, 1995; accepted May 25, 1995 (5T3592). We describe a simple new technique based on the affinity of imidazole and osmium tetroxide for unsaturated lipids. Organs (e.g., kidney, liver, intestine) were perfused in vivo with a glutaraldehyde solution. Tissue fragments were then immersed in a solution containing imidazole and Os04 and are further stained with a double lead and copper citrate solution. Ultra-thin (0.06 pm) or thick (0.1-0.3 pm) sec- tions were observed with transmission electron microscopy (80-100 kV). The method presented permits excellent visu- alization of cell membranes (e.g., endoplasmic reticulum, Introduction The use of en bloc staining coupled with thick sections (0.2-0.5 pm) has provided new views on the spatial organization of cell or- ganelles in the cytoplasm, i.e., mitochondria (I), Golgi apparatus (2,3), and endoplasmic reticulum (4-7). However, metal impreg- nation has a major drawback because it cannot offer clear images of cell membranes and therefore the relationships among organelles cannot be observed with precision. When post-fixed with potas- sium ferrocyanide and osmium (8), cell membranes could be visual- ized in 0.2-0.3-pm sections. However, because the many plasma membrane infoldings act as a barrier to resin penetration, the sec- tions become too fragile to resist the electron beam and are diffi- cult to observe. We present a new method, based on the affinity of imidazole and os04 for unsaturated lipids, that has been al- ready demonstrated by Angermuller and Fahimi (9). This method appears to achieve better resin penetration than osmium/ferro- cyanide and therefore produces excellent contrast for membrane staining. Supported by grant MT-2862 from the Medical Research Council of Canada and by the Minisdie de I'Education of the Government of Qut- bec, Canada. ' Correspondence to: Dr. Michel Bergeron, Dept. de physiologie, Facultt de Medecine. Universite de Montrtal, CP 6128, Succ. Centre-ville, Montreal, Quebec, Canada, H3C 3J7. endocytotic apparatus) because it favors good resin penetra- tion and the alkaline pH preserves cell volume. A better stereomicroscopic analysis of the relationship between cell organelles can be carried out with thick sections. The im- idazolelosmium can be used routinely because the techni- cal steps are easy and simple to follow. Furthermore, it complement other cytochemical methods. (J Hisrochem cyfochem 43:1079-1084, 1995) KEY WORDS: Endoplasmic reticulum; Cytomembranes; Histochemis- try; Kidney; Liver; Rat; Imidazole; Osmium tetroxide.
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This note was uploaded on 05/28/2010 for the course BIOLOGY 03234 taught by Professor Sochacka during the Spring '10 term at Ghent University.

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A Simple Technique for Staining of Cell Membranes with -...

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