Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier. Studies on the synthetizing reaction technique, the indigogenic method, the metal salt method, the post-azo-coupling technique, and the tetrazolium salt technique.
A recently developed method for the (quantitative) demonstration of glucose-6-phosphate dehydrogenase activity in individual cells with the use of a polyacrylamide carrier has been extended for other enzyme cytochemical techniques. Isolated hepatocytes have been incorporated in the matrix of a thin transparent polyacrylamide gel prior to incubation in a cytochemical medium. The techniques which have been applied are the synthetizing reaction technique for glycogen phosphorylase, the indigogenic method for nonspecific esterase, the metal salt method for glucose-6-phosphatase, the post-azo-coupling technique for acid phosphatase, and the tetrazolium salt technique for succinate and lactate dehydrogenase activities. In all cases a few major problems which occur in the cytochemistry on single cells seem to be solved. The morphology is very well preserved, the final reaction product seems to be precipitated at the expected site of enzyme activity and the coloured end-product is highly specific for the enzyme activity to be studied, as has been demonstrated well with control experiments. The conclusion is reached, therefore, that this relatively simple device can be used routinely for the optimalization of enzyme cytochemistry of single cells.
D000180:Acrylic Resins; D000818:Animals; D001391:Azo Compounds; D004396:Coloring Agents; D004798:Enzymes / Q000032:analysis*; D005404:Fixatives; D006651:Histocytochemistry; D008099:Liver / Q000201:enzymology; D008297:Male; D008670:Metals; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D013778:Tetrazolium Salts; D000099166:Polyacrylamides
D000180:Acrylic Resins; D001391:Azo Compounds; D004396:Coloring Agents; D004798:Enzymes; D005404:Fixatives; D008670:Metals; D013778:Tetrazolium Salts; D000099166:Polyacrylamides