A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. I. Optimalization of the staining procedure.
A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual human erythrocytes is described. This staining method can be used for the rapid routine discrimination of patients with a deficiency of the enzyme in its homozygote or heterozygote form, but also for quantitative localization of its activity in individual erythrocytes. The staining procedure in its optimal form consists of a treatment of the erythrocytes with sodium nitrite, then a "fixation" in 0.025% glutaraldehyde (under NADP+ protection of the active site of the enzyme), followed by incubation of the cells in suspension in the presence of tetranitro BT, 1-methoxyphenazine methosulphate and polyvinyl alcohol. Using this new technique, a sharp localization is obtained of the glucose-6-phosphate dehydrogenase activity, which enables discrimination between red cells with different levels of enzyme activity, as a consequence of enzyme deficiencies or age changes.
D004912:Erythrocytes / Q000201:enzymology*; D005954:Glucosephosphate Dehydrogenase / Q000097:blood*; D005976:Glutaral; D006651:Histocytochemistry; D006801:Humans; D008773:Methylphenazonium Methosulfate / Q000031:analogs & derivatives; D009249:NADP; D011142:Polyvinyl Alcohol; D012977:Sodium Nitrite; D013194:Staining and Labeling; D013778:Tetrazolium Salts
D013778:Tetrazolium Salts; C028130:tetranitrotetrazolium blue; C033411:1-methoxyphenazine; D008773:Methylphenazonium Methosulfate; D009249:NADP; D011142:Polyvinyl Alcohol; D005954:Glucosephosphate Dehydrogenase; D012977:Sodium Nitrite; D005976:Glutaral