Dehydrogenase enzyme histochemistry on freeze-dried or fixed resin-embedded tissue.
A method has been developed for the histochemical demonstration of a variety of dehydrogenases in freeze-dried or fixed resin-embedded tissue. Seven dehydrogenases were studied. Lactate dehydrogenase, NADH dehydrogenase and NADPH tetrazolium reductase were all demonstrable in sections of paraformaldehyde-fixed resin-embedded tissue. Freeze-dried specimens were embedded, without fixation, in glycol methacrylate resin or LR Gold resin at either 4 degrees C or -20 degrees C. All the dehydrogenases except succinate dehydrogenase retained their activity in freeze-dried, resin-embedded tissue. Enzyme activity was maximally preserved by embedding the freeze-dried tissue specimens in glycol methacrylate resin at -20 degrees C. The dehydrogenases were accurately localized without any diffusion when the tissue sections were incubated in aqueous media. Addition of a colloid stabilizer to the incubating medium was not required. Freeze-drying combined with low-temperature resin embedding permits accurate enzyme localization without diffusion, maintenance of enzyme activity and excellent tissue morphology.
Murray
G I
GI
Department of Pathology, University of Aberdeen, UK.
Burke
M D
MD
Ewen
S W
SW
D000818:Animals; D005260:Female; D005612:Freeze Drying / Q000379:methods; D006651:Histocytochemistry / Q000379:methods*; D007583:Jejunum / Q000201:enzymology; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D051379:Mice; D008815:Mice, Inbred Strains; D009206:Myocardium / Q000201:enzymology; D010053:Ovary / Q000201:enzymology; D010088:Oxidoreductases / Q000032:analysis*; D010179:Pancreas / Q000201:enzymology; D012116:Resins, Plant; D013154:Spleen / Q000201:enzymology
D012116:Resins, Plant; D010088:Oxidoreductases