A new method for the enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier.
A new method for enzyme cytochemical studies on individual cells is developed. Cells are incorporated in the matrix of a thin film of transparent polyacrylamide prior to incubation in a cytochemical medium. Five different kinds of individual cells, i.e. isolated rat hepatocytes, isolated mouse oocytes, cultivated human fibroblasts, rat thymocytes and human blood cells are used for testing the applicability of this method for the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetranitro BT. The incorporation technique solves at least some of the problems occurring with enzyme cytochemistry on single cells. The morphology of the cell is very well preserved, the formazan precipitation due to enzyme activity occurs entirely within the cell cytoplasm, the nothing dehydrogenase activity can be kept very low and the loss of cells is completely prevented with all cell types used.
D000180:Acrylic Resins; D000818:Animals; D001773:Blood Cells / Q000201:enzymology; D004798:Enzymes / Q000032:analysis*; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D009865:Oocytes / Q000201:enzymology; D051381:Rats; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000201:enzymology
D000180:Acrylic Resins; D004798:Enzymes; C016680:polyacrylamide gels; D005954:Glucosephosphate Dehydrogenase