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Freeze substituted tissue in 5'-nucleotidase histochemistry. Comparative histochemical and biochemical investigations

K Klaushofer and H von Mayersbach

The suitability of freeze-substitution in n-butanol and paraffin embedding of tissues for the histochemical demonstration of 5'- nucleotidase was investigated and compared with commonly used preparation techniques, such as fresh frozen sections and cryostate sections of cold formalin and glutaraldehyde-fixed rat liver. The influences of each step of the preparation techniques on the enzyme activity were controlled by a quantitative radiochemical assay. Freeze substitution was revealed to be superior to the commonly used preparation techniques with respect to: 1) high sensitivity and specificity of the histochemical 5'-nucleotidase reaction (this is based on the fact that incubation media with very low lead concentrations (0,6 mM/1) can be used); 2) excellent morphological appearance of the tissues showing cytological details of enzyme localization; 3) unlimited storage of the tissue materials and ease of sectioning.

Volume 27, Issue 12, pp. 1582-1587, 12/01/1979
Copyright © 1979 by The Histochemical Society


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T. Takizawa
5'-Nucleotidase in Rat Photoreceptor Cells and Pigment Epithelial Cells Processed by Rapid-freezing Enzyme Cytochemistry
J. Histochem. Cytochem., September 1, 1998; 46(9): 1091 - 1096.
[Abstract] [Full Text]



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