Summary
The effects of heavy metal salt staining procedures on the reaction products obtained in the demonstration of arylsulphatase and of acid phosphatase were studied.
Lead citrate staining at pH 12 was found to cause a very marked dissolution of barium sulphate and a moderate dissolution of lead sulphate. The staining with uranyl acetate was found to dissolve moderately both barium and lead sulphate.
Neither lead citrate nor uranyl acetate staining had any remarkable effect on lead phosphate.
The mechanism of the dissolution and the possibilities to avoid it were discussed.
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Arstila, A. U., S. Jaakkola, H. Kalimo, H. Helminen, and V. K. Hopsu-Havu: Electron diffraction as the control of enzyme histochemical reactions at the ultrastructural level. J. Microscopie 5, 777–780 (1966).
Barrnett, R. J., and G. E. Palade: Enzymatic activity in the M band. J. biophys. biochem. Cytol. 6, 163–169 (1959).
Birks, R. I., and C. M. Brown: A method for locating the cholinesterase of a mammalian myoneural junction by electron microscopy. J. Physiol. (Lond.) 152, 5P (1960).
Brandes, J., H. Zetterquist, and H. Sheldon: Histochemical techniques for electron microscopy: Alkaline phosphatase. Nature (Lond.) 177, 382–383 (1956).
Ericsson, J. L. E., and B. F. Trump: Observations on the application to electron microscopy of the lead phosphate technique for the demonstration of acid phosphatase. Histochemie 4, 470–487 (1965).
Essner, E., and A. B. Novikoff: Acid phosphatase activity in hepatic lysosomes: Electron microscopic demonstration of its reaction product. J. Histochem. Cytochem. 8, 318 (1960).
—, and B. Masch: Adenosine triphosphatase and 5-nucleotidase activity in the plasma membrane of liver cells as revealed by electron microscopy. J. biophys. biochem. Cytol. 4, 711–716 (1958).
Goldfischer, S.: The cytochemical demonstration of lysosomal aryl sulphatase activity by light and electron microscopy. J. Histochem. Cytochem. 13, 520–524 (1965).
Holt, S. J., and R. M. Hicks: 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 (1961).
Hopsu, V. K., A. Arstila, and G. G. Glenner: A method for electromicroscopic localization of aryl-sulphatase. Ann. Med. exp. Fenn. 43, 114–116 (1965a).
—: The electron microscopic localization of aryl sulphatase activity. J. Histochem. Cytochem. 13, 711–712 (1965b).
— H. J. Helminen, H. O. Kalimo, and G. G. Glenner: Improvements in the method for the electronmicroscopic localization of arylsulphatase activity. Histochemie 8, 54–64 (1967).
Kalimo, H. O., H. Helminen, A. Arstila, and V.K. Hopsu-Havu: Methodical observations on the demonstration of hydrolytic enzymes at electronmicroscopic level. Scand. J. clin. Lab. Invest. 19, Suppl. 95, 77 (1967).
Lewis, P. R., and C. C. D. Shute: Demonstration of cholinesterase activity with the electronmicroscopy. J. Physiol. (Lond.) 175, 5–7P (1964).
Luft, J. H.: Improvements in epon resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961).
Mitsutani, A., and R. J. Barrnett: Fine structural demonstration of phosphatase activity at pH 9. Nature (Lond.) 206, 1001 (1965).
Mölbert, E., F. Duspiva, and O. v. Beimling: The demonstration of alkaline phosphatase in the electron microscope. J. biophys. biochem. Cytol. 7, 387–390 (1960).
Nelson, L.: Cytochemical studies with the electron microscope. I. Adenosine triphosphatase in rat spermatozoa. Biochem. biophys. Acta (Amst.) 27, 634–641 (1958).
Reynolds, E. S.: The use of lead citrate as an electron dense stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).
Sheldon, H., H. Zetterquist, and D. Brandes: Histochemical reactions for electron miroscopy: Acid phosphatase. Exp. Cell Res. 9, 592–596 (1955).
Tice, L.W., and R. J. Barrnett: The fine structural localization of glucose-6-phosphatase in rat liver. J. Histochem. Cytochem. 10, 754–762 (1962).
Torack, R. M., and R. J. Barrnett: The fine structural localization of nucleoside phosphatase in the blood brain barrier. J. Neuropath. exp. Neurol. 23, 46–59 (1964).
Venable, J. H., and R. Coggeshall: A simplified lead citrate stain for use in electron microscopy. J. Cell Biol. 25, 407 (1965).
Watson, M. L.: Staining of tissue sections for electron microscopy with heavy metals. J. biophys. biochem. Cytol. 4, 475–478 (1958).
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Kalimo, H.O., Helminen, H.J., Arstila, A.U. et al. The loss of enzyme reaction products from ultrathin sections during the staining for electron microscopy. Histochemistry Histochimie 14, 123–130 (1968). https://doi.org/10.1007/BF00268039
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DOI: https://doi.org/10.1007/BF00268039