Synopsis
A model system developed for the study of the dynamics of capture reactions for diffusable compounds in cytochemistry served as a basis for the experiments reported in the present paper. The model was used to study the effect of the composition of the cytochemical medium on the trapping of phosphate ions by lead (II) ions in acid phosphatase cytochemistry. In this system a phosphate-containing solution and a lead-containing solution (cytochemical medium) are pumped along opposite sides of a polyacrylamide film. The phosphate concentration at which measurable precipitation starts in the film (critical phosphate concentration) was taken as a measure of the trapping efficiency of the cytochemical medium. The addition of β-glycerophosphate and cytidine-5′-monophosphate to a buffered lead-containing solution resulted in a higher critical phosphate concentration. Both substrates had an effect on the crystal form of lead phosphate. The addition of chloride ions and acetone, as well as decreasing the molarity of the acetate buffer of the cytochemical medium, were found to lower the critical phosphate concentration, whereas the addition of fluoride ions, glucose, and sucrose had no effect. From the effect of variations in the composition of the cytochemical medium on the trapping efficiency and the turnover number of acid phosphatase in the medium, it was possible to predict which cytochemical medium would be the most suitable for the demonstration of acid phosphatase activity in guinea-pig peritoneal exudate cells. The results were in accordance with the localization of acid phosphatase activity: the higher the trapping efficiency and the turnover number, the higher the amount of precipitate and the number of positive enzymatic sites. In this way an improved cytochemical medium for acid phosphatase was developed.
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References
Altman, H. J., Fürstenau, E., Gielewski, A. &Scholz, L. (1971). Photometrische Bestimmung kleiner Phosphatmengen mit Malachitgrün.J. Anal. Chem. 256, 274–6.
Bainton, D. F. &Farquhar, M. G. (1968). Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. II. Cytochemistry and electron microscopy of bone marrow cells.J. Cell Biol. 39, 299–317.
Barka, T. &Anderson, P. J. (1962). Histochemical methods for acid phosphatase using hexazonium patarosanilin as coupler.J. Histochem. Cytochem. 10, 741–53.
Berg, G. G., Lyon, D. &Campbell, M. (1972). Faster histochemical reaction for adenosine triphosphatase in presence of chloride salts, with studies on the mechanism of precipitation.J. Histochem. Cytochem. 20, 39–55.
Brunk, U. T. &Ericsson, J. L. E. (1972). The demonstration of acid phosphatase inin vitro cultured tissues cells. Studies on the significance of fixation, tonicity and permeability.Histochem. J. 4, 349–63.
Burnes, E. A. &Hume, D. N. A. (1956). Acetato complexes of lead in aqueous solution.J. Am. Chem. Soc. 78, 3958–62.
Butterworth, J. (1971). Histochemistry of non-specific phosphatases: the use ofp-nitrophenyl phosphate and beta-glycerophosphate as substrates.Histochem. J. 3, 477–87.
Carmody, W. R. (1959). Variation of the solubility product constant with ionic strength.J. Chem. Educ. 36, 125–7.
Cornelisse, C. J., Hermens, W. Th., Tjok Joe, M., Duijndam, W. A. L. &Van Duijn, P. (1976). A theoretical study of concentration profiles of primary cytochemical-enzyme reaction products in membrane-bound cell organelles and its application to lysosomal acid phosphatase.Histochem. J. 8, 609–24.
Cornelisse, C. J. &Van Duijn, P. (1974). A new method for the investigation of the capture reaction in phosphatase cytochemistry. III. Effects of the composition of the incubation medium on the trapping of phosphate ions in a model system.J. Histochem. Cytochem. 22, 110–9.
Daems, W. Th. &Brederoo, P. (1973). Electron microscopical studies on the structure, phagocytic properties, and peroxidatic activity of resident and exudate peritoneal macrophages in the guinea-pig.Z. Zellforsch. 144, 247–97.
Daems, W. Th., Wisse, E, &Brederoo, P. (1969). Electron microscopy of the vacuolar apparatus. In:Lysosomes in biology and pathology (eds. J. T. Dingle and H. B. Fell) Vol. 1, pp. 64–112. Amsterdam and London: North-Holland.
De Jong, A. S. H., Hak, T. J., Van Duijn, P. &Daems, W. Th. (1978). A new dynamic model system for the study of capture reactions for diffusable compounds in cytochemistry. I. Description of the model with special attention to phosphate capture in acid phosphatase cytochemistry.J. Histochem. Cytochem. 26, 331–9.
De Jong, A. S. H., Van Duijn, P. &Daems, W. Th. (1976). Cytochemical model system for microsomal rat liver glucose-6-phosphatase.J. Histochem. Cytochem. 24, 643–51.
Ericsson, J. L. E. &Trump, B. F. (1965). Observations on the application to electron microscopy of the lead phosphate technique for the demonstration of acid phosphatase.Histochemie 4, 470–87.
Etherton, J. E. &Botham, C. M. (1970). Factors affecting lead capture methods for the fine localization of rat lung acid phosphatase.Histochem. J. 2, 507–19.
Gomori, G. (1939). Microtechnical demonstration of phosphatase in sections.Proc. Soc. Exp. Biol. Med. 42, 23.
Gomori, G. (1952).Microscopic Histochemistry, pp. 189–194. Chicago: University of Chicago Press.
Goodman, R. C. &Petrucci, R. H. (1965). Is the solubility product constant?J. Chem. Educ. 42, 104–5.
Holt, S. J. &Hicks, M. (1961). The localization of acid phosphatase in rat liver cells as revealed by combined cytochemical staining and electron microscopy.J. Biophys. Biochem. Cytol. 11, 47–66.
Jowett, M. &Price, H. I. (1932). Solubilities of the phosphates of lead.Trans. Farady Soc. 28, 668–81.
Lake, B. D. (1966). The histochemistry of phosphatases: the use of lead acetate instead of lead nitrate.J. Roy. Microsc. Soc. 85, 73–5.
Lazarus, S. S., Volk, B. W. &Barden, H. (1966). Localization of acid phosphatase activity and secretion mechanism in rabbit pancreatic B-cells.J. Histochem. Cytochem. 14, 233–46.
Luppa, H., Weiss, J. &Martin, G. (1972). Eine Beitrag zur elektron-enmikroskopisch-histochemischen. Darstellung der unspezifischen sauren Phosphatase mitp-Nitrophenylphosphat.Acta Histochem. 43, 184–8.
Millonig, G. (1961). A modified procedure for lead staining of thin sections.J. Biophys. Biochem. Cytol. 11, 736–9.
Miyayama, H., Solomon, R., Sasaki, M., Lin, C. W. &Fishman, W. H. (1975). Demonstration of lysosomal and extra-lysosomal sites for acid phosphatase in mouse kidney tubule cells withp0nitrophenyl phosphate lead-salt technique.J. Histochem. Cytochem. 23, 439–51.
Novikoff, A. B. (1963). Lysosomes in the physiology and pathology of cells: contributions of staining methods. In:Ciba Foundation Symposium Lysosomes (eds. A. U. S. de Reuck and M. P. Cameron) pp. 36–77. London: Churchill.
Pfeifer, U. &Witschel, H. (1972). Zum zeitlichen Ablauf der für die Elektronenmikroskopie modifizierten Gomori-Reaktion auf saure Phosphatase. Quantitative Bestimmung des Reaktionsproduktes mit der Atomabsorptionspektrometrie.Acta Histochem., Suppl. Bd.XII, 111–20.
Robertson, W. G. (1973). Factors affecting the precipitation of calcium phosphatein vitro.Calc. Tiss. Res. 11, 311–22.
Ryder, T. A. &Bowen, I. D. (1975). A method for the fine structural localization of acid phosphatase activity usingp-nitrophenyl phosphate as substrate.J. Histochem. Cytochem. 23, 235–7.
Smith, R. E. &Farquhar, M. G. (1966). Lysosome function in the regulation of the secretory process in cells of the anterior pituitary gland.J. Cell Biol. 31, 319–47.
Takamatsu, H. (1939). Histologische und biochemische Untersuchungsmethodik der Phosphatase und deren Verteilung in verschiedenen Organen und Geweben.Trans. Soc. Path. Jap. 29, 492.
Takemori, S., Furuya, E., Suzuki, H. &Katagiri, M. (1967). Stabilization of enzyme activity by an organic solvent.Nature 215, 417–9.
Tandler, C. J. (1953). The use of cobalt acetate in the histochemical technic for acid phosphatase.J. Histochem. Cytochem. 1, 151–3.
Termine, J. D., Peckauskas, R. A. &Posner, A. S. (1970). Calcium phosphate formationin vitro. II. Effects of environment on amorphous-crystalline transformation.Arch. Biochem. Biophys. 140, 318–25.
Termine, J. D. &Posner.A. S. (1970). Calcium phosphate formationin vitro. I. Factors affecting initial phase separation.Arch. Biochem. Biophys. 140, 307–17.
Weissenfels, N. (1967). Nachweis von saurer Phosphatase in gezüchteten Hühnerherzmyoblasten.Histochemie 9, 189–202.
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De Jong, A.S.H., Hak, T.J., Van Duijn, P. et al. A new dynamic model system for the study of capture reactions for diffusable compounds in cytochemistry. II. Effect of the composition of the incubation medium on the trapping of phosphate ions in acid phosphatase cytochemistry. Histochem J 11, 145–161 (1979). https://doi.org/10.1007/BF01002992
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DOI: https://doi.org/10.1007/BF01002992