Summary
The new method, utilizing “lead citrate” as capture reagent, for the ultracytochemical demonstration of non-specific alkaline phosphatase activity at high alkaline ranges of pH was introduced. Results obtained by the new “lead citrate” method in various tissues of the rat were presented and discussed in connection with observations made by other methods. The method can be used at the light microscopic level also.
Similar content being viewed by others
Bibliography
Bakay, L.: The Blood-Brain Barrier, with special regard to the use of radioactive isotopes. Springfield (Ill.): Ch. C. Thomas 1956.
Brandes, D., H. Zetterqvist, and H. Sheldon: Histochemical techniques for electron microscopy: Alkaline phosphatase. Nature (Lond.) 177, 382–383 (1956).
Byczkowska-Smyk, W., et W. Bernhard: Essais de cytochimie ultrastructurale. Recherche de la phosphatase alcaline dans le rein du rat à l'aide du microscope électronique. C. R. Acad. Sci. (Paris) 251, 3085–3087 (1960).
Caulfield, J. B.: Effects of varying the vehicle for OsO4 in tissue fixation. J. biophys. biochem. Cytol. 3, 827–830 (1957).
Chase, W. H.: The demonstration of alkaline phosphatase activity in frozen-dried mouse gut in the electron microscope. J. Histochem. Cytochem. 11, 96–101 (1963).
Clark jr., S. L.: The localization of alkaline phosphatase in tissues of mice, using the electron microscope. Amer. J. Anat. 109, 57–83 (1961).
Goldfischer, S., E. Essner, and A. B. Novikoff: The localization of phosphatase activities at the level of ultrastructure. J. Histochem. Cytochem. 12, 72–95 (1964).
Hoff, H. F., and J. Graf: An electron microscopy study of phosphatase activity in the endothelial cell of rabbit aorta. J. Histochem. Cytochem. 14, 719–724 (1966).
Hugon, J., and M. Borgers: A direct lead method for the electron microscopic visualization of alkaline phosphatase activity. J. Histochem. Cytochem. 14, 429–431 (1966a).
—: Ultrastructural localization of alkaline phosphatase activity in the absorbing cells of the duodenum of mouse. J. Histochem. Cytochem. 14, 629–640 (1966b).
Luft, J. H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961).
Marchesi, V. T., and R. J. Barrnett: The localization of nucleosidephosphatase activity in different types of small blood vessels. J. Ultrastruct. Res. 10, 103–115 (1964).
Mayahara, H., T. Saito, H. Hirano, and K. Ogawa: Nucleoside phosphatases demonstrated by the lead citrate method at high alkaline pH (in preparation).
Millington, P. F., and A. C. Brown: Electron microscope studies of the distribution of phosphatase in rat intestinal epithelium from birth to ten days after weaning. Histochemie 8, 107–121 (1967).
Mizutani, A., and R. J. Barrnett: Fine structural demonstration of phosphatase activity at pH 9. Nature (Lond.) 206, 1001–1003 (1965).
Mölbert, E. R. G., F. Duspiva, and O. H. Von Deimling: The demonstration of alkaline phosphatase in the electron microscope. J. biophys. biochem. Cytol. 7, 387–390 (1960).
Nagano, T.: Fine structural localization of phosphatase activities in the proximal convoluted tubule of the rat. Okajimas Folia anat. jap. 40, 647–661 (1965).
Ogawa, K., T. Saito, H. Hirano, and H. Mayahara: Automicrochopper model KO-1 for making non-frozen sections [In Japanese]. 7th Ann. Meet. Jap. Soc. Histoand Cyto-chem., Chiba, Nov. 7 (1966).
—, Y. Shinonaga, and T. Suzuki: Metallophilia of the striated border of the rat jejunal epithelial cells. Acta anat. Nippon. 37, 134–139 (1962).
Reale, E.: Electron microscopic localization of alkaline phosphatase from material prepared with the cryostat-microtome. Exp. Cell Res. 26, 210–228 (1962).
—, and L. Luciano: A probable source of errors in electronhistochemistry. J. Histochem. Cytochem. 12, 713–715 (1964).
Reale, E., u. L. Luciano: Kritische elektronenmikroskopische Studien über die Lokalisation der Aktivität alkalischer Phosphatase im Hauptstück der Niere von Mäusen. Histochemie 8, 302–314 (1967).
Reynolds, E. S.: The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–213 (1963).
Robertson, J. D., T. S. Bodenheimer, and D. E. Stage: The ultrastructure of Mauthner cell synapses and node in goldfish brains. J. Cell Biol. 19, 159–199 (1963).
Sabatini, D. D., K. Bensch, and R. J. Barrnett: Cytochemistry and electron microscopy. The preservation of cellular utrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17, 19–58 (1963).
Saito, T., and K. Ogawa: Ultracytochemical demonstration of fructose-1,6-diphosphatase (D-fructose-1,6-diphosphate 1-phosphohydrolase) activity in the rat liver using lead citrate as capture reagent. J. Ultrastr. Res., in press.
Shinonaga, Y., Y. Kondo, K. Ogawa, and S. Ishii: Alkaline phosphatase activity in normal rodent brain and experimentally induced cerebral swelling. Arch. histol. jap. 22, 193–201 (1962).
Smith, R. E., and M. G. Farquhar: Preparation of nonfrozen sections for electron microscope cytochemistry. RCA Scient. Instr. News 10, 13–18 (1965).
Thé, G. De: Méthode au plomb pour la mise en évidence de la phosphatase alcaline en microscopie électronique. J. Microscopie 4, 130–131 (1965).
Torack, R. M., and R. J. Barrnett: Nucleoside phosphatase activity in membranous fine structures of neurons and glia. J. Histochem. Cytochem. 11, 763–772 (1963).
Tranzer, J. P.: Utilisation de citrate de plomb pour la mise en évidence de la phosphatase alcaline au microscope électronique. J. Microscopie 4, 409–412 (1965).
Wachstein, M., and M. Besen: Electron microscopic localization of phosphatase activity in the brush border of the rat kidney. J. Histochem. Cytochem. 11, 447 (1963).
Author information
Authors and Affiliations
Additional information
Supported by grants from the U.S. Public Health Service MH 12269-01, the Jane Coffin Childs Memorial Fund for Medical Research (Project No. 196), the Anna Fuller Fund and the Department of Education, the Japanese Government (Sogokenkyu UDC 611.015.3:611-018:576.311).
Rights and permissions
About this article
Cite this article
Mayahara, H., Hirano, H., Saito, T. et al. The new lead citrate method for the ultracytochemical demonstration of activity of non-specific alkaline phosphatase (orthophosphoric monoester phosphohydrolase). Histochemie 11, 88–96 (1967). https://doi.org/10.1007/BF00326615
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00326615