A polychromatic staining method for epoxy embedded tissue: a new combination of methylene blue and basic fuchsine for light microscopy.
A simple and rapid method is described for staining semithin sections of material embedded in epoxy resin for observing tissues prior to transmission electron microscopy. The method is suitable for tissue fixed with a glutaraldehyde-formaldehyde mixture and postfixed in osmium tetroxide. No etching or oxidizing procedures are necessary. Sections 0.5-0.8 microm thick are dried onto a slide and stained with either 0.75% methylene blue and 0.25% azure B or 0.5% methylene blue and 0.5% azure II in 0.5% aqueous borax and heated over a flame for 8-10 sec. The slides are rinsed with water, then stained the same way with 0.1% basic fuchsine in 5% aqueous ethanol. Cytoplasm stains blue; nuclei darker blue; collagen, mucus and elastin pink to red; fat and intracellular lipid droplets gray-green.
D'Amico
F
F
CMEM, Medical Electron Microscopy Unit, University of Catania, via Androne 87/a 95129, Catania, Italy. f.damico@unict.it
D000818:Animals; D001399:Azure Stains; D004853:Epoxy Resins; D005404:Fixatives; D006801:Humans; D007422:Intestines / Q000166:cytology; D008297:Male; D008751:Methylene Blue; D051379:Mice; D008853:Microscopy; D008854:Microscopy, Electron / Q000379:methods; D008867:Microtomy; D016613:Plastic Embedding; D011108:Polymers; D011651:Pulmonary Artery / Q000166:cytology; D012394:Rosaniline Dyes; D012867:Skin / Q000166:cytology; D013194:Staining and Labeling / Q000379:methods*; D013552:Swine; D013737:Testis / Q000166:cytology; D016707:Tissue Fixation
D001399:Azure Stains; D004853:Epoxy Resins; D005404:Fixatives; D011108:Polymers; D012394:Rosaniline Dyes; C025485:basic fuchsin; C025819:Azure B; D008751:Methylene Blue