Freeze-drying of unfixed monolayer cultures for electron microscopic autoradiography

A method has been developed for freezing, drying and embedding of unfixed monolayer cultures for electron microscopic autoradiography (EM ARG). Experimental results showed: a) Aclar 33 C was a more suitable substrate than the plastic of petri dishes, b) cultures pressed rapidly against the polished...

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Bibliographic Details
Published inHistochemistry (Berlin) Vol. 58; no. 4; p. 259
Main Authors Burry, R W, Lasher, R S
Format Journal Article
LanguageEnglish
Published Germany 13.12.1978
Subjects
Animals
Autoradiography - methods
Cerebellum
Culture Techniques
Fixatives
Freeze Drying - methods
gamma-Aminobutyric Acid - isolation & purification
Microscopy, Electron
Neurons - analysis
Rats
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Summary:A method has been developed for freezing, drying and embedding of unfixed monolayer cultures for electron microscopic autoradiography (EM ARG). Experimental results showed: a) Aclar 33 C was a more suitable substrate than the plastic of petri dishes, b) cultures pressed rapidly against the polished face of a large copper cylinder chilled in liquid nitrogen had better cellular morphology than did cultures dipped in Freon 12 chilled in liquid nitrogen, and c) cultures embedded in Epon alone had finer extracellular ice spaces and lower background grain densities than did cultures embedded in Epon with 1% silicone. This method has been used to evaluate the effect of fixation on the localization of the neurotransmitter, 3H-gamma-aminobutyric acid (3H-GABA), in neurons of dispersed cell cultures. EM ARG results showed that the neuronal cell bodies and vesicle elements were present in similar numbers in both glutaraldehyde fixed and freeze-dried cultures.
ISSN:0301-5564
DOI:10.1007/BF00495382