A simple technique for staining of cell membranes with imidazole and osmium tetroxide.
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 OsO4 and are further stained with a double lead and copper citrate solution. Ultra-thin (0.06 microns) or thick (0.1-0.3 microns) sections were observed with transmission electron microscopy (80-100 kV). The method presented permits excellent visualization of cell membranes (e.g., endoplasmic reticulum, endocytotic apparatus) because it favors good resin penetration 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 imidazole/osmium can be used routinely because the technical steps are easy and simple to follow. Furthermore, it can complement other cytochemical methods.
Thiéry
G
G
Department of Physiology, Université de Montréal, Québec, Canada.
Bernier
J
J
Bergeron
M
M
D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure*; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005404:Fixatives; D005976:Glutaral; D007093:Imidazoles; D007202:Indicators and Reagents; D007422:Intestines / Q000648:ultrastructure; D007668:Kidney / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000648:ultrastructure; D008099:Liver / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron / Q000379:methods; D009399:Nephrons / Q000648:ultrastructure; D009993:Osmium Tetroxide; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling / Q000379:methods
D005404:Fixatives; D007093:Imidazoles; D007202:Indicators and Reagents; C029899:imidazole; D009993:Osmium Tetroxide; D005976:Glutaral