Semipermeable membranes for improving the histochemical demonstration of enzyme activities in tissue sections. VI. D-glucose 6-phosphate isomerase and phosphoglucomutase

Abstract

Improved histochemical multi-step techniques for the demonstration of glucose 6-phosphate isomerase and phosphoglucomutase in tissue sections are described. With these techniques a semipermeable membrane is interposed between the incubating solutions and the tissue sections preventing diffusion of enzymes into the medium during incubation. In the histochemical system the glucosephosphate isomerase converts the substrate D-fructo-furanose 6-phosphoric acid to D-gluco-pyranose 6-phosphoric acid, and the phosphoglucomutase converts the substrate alpha-D-glucose 1-phosphate to the same reagent, which in turn is oxidized, by exogenous and endogenous glucose 6-phosphate dehydrogenase to D-glucono-delta-lactone 6-phosphoric acid. Concomittantly the electrons are transferred via NADP+, phenazine methosulphate and menadione to nitro-BT. Sodiumazide and amytal are incorporated to block electron transfer to the cytochromes.