Summary
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 cells 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.
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References
Biggers JD, Whittingham DG, Donahue RP (1967) The pattern of energy metabolism in the mouse oocyte and zygote. Proc Natl Acad Sci USA 58:560–567
James J (1977) The genesis of polyploidy in rat liver parenchymal cells. Cytobiologie 15:410–419
Knook DL, Sleyster EC, Teutsch HF (1980) High activity of glucose-6-phosphate dehydrogenase in Kupffer cells isolated from rat liver. Histochemistry 69:211–216
Löhr GW, Waller HD (1970) Glucose-6-phosphat-Dehydrogenase. In: Bergmeyer HU (ed) Methoden der Enzymatischen Analyse, Vol 1, 2nd edn. Verlag Chemie, Weinheim, pp 599–606
Papadimitriou JM, Van Duijn P (1970) Effecfts of fixation and substrate protection on the isoenzymes of aspartate aminotransferase studied in a quantitative cytochemical model system. J Cell Biol 47:71–83
Pette D, Wimmer M (1979) Kinetic microphotometric activity determination in enzyme containing gels and model studies with tissue sections. Histochemistry 64:11–22
Pollak A, Blumenfeld H, Wax M, Baughn RL, Whitesides GM (1980) Enzyme immobilization by condensation copolymerization into cross-linked polyacrylamide gels. J Am Chem Soc 102:6324–6335
Rieder H, Teutsch HF, Sasse D (1978) NADP-dependent dehydrogenases in rat liver parenchyma. I. Methodological studies on the qualitative histochemistry of G6PDH, 6PGDH, malic enzyme and ICDH. Histochemistry 56:283–298
Roholl P, Hogenes T, Leene W (1977) A model for the development of a rabbit T cell surface marker. In: Solomon JB, Horton JD (eds) Developmental immunobiology. Elsevier North-Holland, Amsterdam, pp 163–170
Stuart J, Gordon PA, Lee TR (1975) Enzyme cytochemistry of blood and marrow cells. Histochem J 7:471–487
Tzortzatou F, Hayhoe FGJ (1974) Cytochemical demonstration of dihydrofolate reductase activity. Br J Haematol 28:199–208
Van der Ploeg M (1975) Quantitative model film studies: a link between enzyme cytochemistry and biochemistry. Acta Histochem (Suppl) 14:69–77
Van Duijn P (1974) Fundamental aspects of enzyme cytochemistry. In: Wisse E, Daems W Th, Molenar I, Van Duijn P (eds) Electron microscopy and cytochemistry. Elsevier North-Holland, Amsterdam, pp 3–23
Van Duijn P (1976) Prospects for microscopical cytochemistry. Histochem J 8:653–676
Van Duijn P, Van der Ploeg M (1980) Microscopic cytochemistry as matrix chemistry. In: Evered D, O'Connor M (eds) Trends in enzyme histochemistry and cytochemistry. Elsevier North-Holland, Amsterdam, pp 209–229
Van Noorden CJF, Tas J (1980) Quantitative aspects of the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetranitro BT studied in a model system of polyacrylamide films. Histochem J 12:669–685
Van Noorden CJF, Tas J (1981a) Model film studies in enzyme histochemistry with special reference to glucose-6-phosphate dehydrogenase. Histochem J 13:187–206
Van Noorden CJF, Tas J (1981b) Advantages of 1-methoxyPMS as electron carrier in dehydrogenase cytochemistry. Proc JR Microsc Soc 16:248
Van Noorden CJF, Tas J (1982a) The role of exogenous electron carriers in NAD(P)-dependent dehydrogenase cytochemistry studied in vitro and with a model system of polyacrylamide films. J Histochem Cytochem 30:12–20
Van Noorden CJF, Tas J (1982b) Advantages of 1-methoxyPMS as electron carrier in dehydrogenase cytochemistry. Histochem J 14: in press
Van Noorden CJF, Tas J, Sanders JAH (1981) Quantitative aspects of the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetrazolium salts studied in a model system of polyacrylamide films. Acta Histochem (Suppl) 24:231–236
Weiss G, Vergani G, Pentz S (1979) Histochemical demonstration of G6PDH- and SDH-activity on cultured fibroblastoid cells from rat pancreas using a “membrane technique”. Histochemistry 64:111–114
Zimmermann H, Pearse AGE (1959) Limitations in the histochemical demonstration of pyridine nucleotide-linked dehydrogenases (“nothing dehydrogenase”). J Histochem Cytochem 7:271–275
Zimmerman N, Geyer G (1981) A polyacrylamide gel method for the cytochemical demonstration of glucose-6-phosphate dehydrogenase activity in mouse sperm. Acta Histochem 68:277–230
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van Noorden, C.J.F., Tas, J., Vogels, I.M.C. et al. A new method for the enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier. Histochemistry 74, 171–181 (1982). https://doi.org/10.1007/BF00495828
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DOI: https://doi.org/10.1007/BF00495828