The use of unfixed cryostat sections for electron microscopic study of D-amino acid oxidase activity in rat liver

Unfixed cryostat sections of rat liver were incubated to demonstrate D-amino acid oxidase activity at the ultrastructural level. Incubation was performed by mounting the sections on a semipermeable membrane which was stretched over a gelled incubation medium containing D-proline as substrate and cer...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1992-12, Vol.40 (12), p.1975-1979
Main Authors: Schellens, JP, Frederiks, WM, Van Noorden, CJ, Vreeling-Sindelarova, H, Marx, F, McMillan, PJ
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Biology
Cryoultramicrotomy - methods
D-Amino-Acid Oxidase - metabolism
Fundamental and applied biological sciences. Psychology
Life Sciences & Biomedicine
Liver - enzymology
Liver - ultrastructure
Liver. Bile. Biliary tracts
Male
Microscopy, Electron - methods
Rats
Rats, Wistar
Science & Technology
Vertebrates: digestive system
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Summary:Unfixed cryostat sections of rat liver were incubated to demonstrate D-amino acid oxidase activity at the ultrastructural level. Incubation was performed by mounting the sections on a semipermeable membrane which was stretched over a gelled incubation medium containing D-proline as substrate and cerium ions as capture reagent for hydrogen peroxide. After an incubation period of 30 min, ultrastructural morphology was retained to such an extent that the final reaction product could be localized in peroxisomes, whereas the crystalline core remained unstained. Control incubations were performed in the absence of substrate; the lack of final reaction product in peroxisomes indicated the specificity of the reaction. We conclude that the semipermeable membrane technique opens new perspectives for localization of enzyme activities at the ultrastructural level without prior tissue fixation, thus enabling localization of the activity of soluble and/or labile enzymes.
ISSN:0022-1554
1551-5044
DOI:10.1177/40.12.1360483