The Use of Formalin Fixation in the Cytochemical Demonstration of Succinic and DPN- and TPN-dependent Dehydrogenases in Mitochondria

Brief formalin fixation in the cold prior to histochemical assay of rat liver and pancreas for various dehydrogenases has been used successfully to circumvent the structural damage and enzymatic loss to which mitochondria of frozen sections would otherwise be subject. To obtain an optimal result a single set of conditions has been devised, including fixation prior to freezing of minute (finely diced) organ blocks in graded concentrations (0.7 to 2.0 per cent) of formaldehyde in chilled (1-4 degrees C) Hanks' balanced salt solution, freezing at not higher than -70 degrees C, and use of nitro-BT or, preferably, tetranitro-BT. The present histochemical study of hepatic and acinar cells indicates that not only are succinic and D-beta-hydroxybutyric dehydrogenases located exclusively in the mitochondria but so are lactic, malic, and the isocitric dehydrogenases.