A simple technique for flat osmicating and flat embedding of immunolabelled vibratome sections of the rat spinal cord for light and electron microscopy.
We describe here a simple technique for flat-osmicating and flat-embedding of immunolabelled vibratome sections. The technique is particularly useful for large specimens such as whole cross-sections of rat spinal cord. After the vibratome section has been flat-osmicated on a flat surface under a glass coverslip, it is dehydrated and then embedded by placing it in a small drop of epoxy resin on a flat base of polypropylene plastic and overlaying a small square piece of cellulose acetate cut from heat-resistant overhead projector transparency film for photocopying. The thin layer of resin containing the flat-embedded vibratome section is then separated from the base and glued onto the flat end of a pre-polymerised blank block of resin. The method produces flat-embedded vibratome sections and thus allows serial large uniformly labelled semithin and ultrathin sections to be obtained of the whole cross-section of the rat spinal cord. This facilitates the observation and quantification of labelled cells in the specimen. Because of its simplicity the technique also allows one worker to process more than 100 vibratome sections at the one time.
Nguyen
K B
KB
Department of Medicine, University of Queensland, Royal Brisbane Hospital, Brisbane, Australia.
Pender
M P
MP
D000818:Animals; D002482:Cellulose / Q000031:analogs & derivatives; D004853:Epoxy Resins; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D008867:Microtomy / Q000379:methods*; D009993:Osmium Tetroxide; D016613:Plastic Embedding / Q000379:methods*; D011126:Polypropylenes; D051381:Rats; D013116:Spinal Cord / Q000033:anatomy & histology* / Q000378:metabolism / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*
D004853:Epoxy Resins; D011126:Polypropylenes; C005062:acetylcellulose; D002482:Cellulose; D009993:Osmium Tetroxide