Summary
The cell coat picture effect which is usually obtained with the conventional RR method, that is with the RR/OsO4 coupled reaction, is investigated. In this first paper, each of conceivable events which might take place between RR, OsO4 and cell surface membrane is discussed or studied. Various tests are carried out on ascites Ehrlich carcinoma cells and Zajdela ascites hepatoma cells treated with numerous chemical reagents, as also on a few pure proteins. The set of data supports the concept that the staining pattern is due to the combination in surface membranes of RR with a “colloidal-like” form of OsO2. The latter might occur during the formation of stable cyclic osmic acid diesters between OsO4 and membrane unsaturated lipids. A possibility by which the resulting marker is though also to be in a “colloidal-like” state is put forward. A next report will deal with this problem.
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References
Bonneau, H., Césarini, J. P.: Proc. 4th European Reg. Conf. Electron Microscopy, Rome, p. 577 (1968)
Bonner, J.: The chemistry and physiology of the pectins. Botan. Rev. 2, 475–497 (1936)
Collin, R. J., Griffith, W. P.: Mechanism of tissue component staining by osmium tetroxide. J. Histochem. Cytochem. 22, 992–996 (1974)
Cook, G. M. W., Stoddart, R. W.: The structure of plasma membranes. In: Surface carbohydrates of the eukaryotic cell, pp. 1–55. London-New York: Academic Press 1973
Criegee, R.: Osmiumsäure-Ester als Zwischenprodukte bei Oxydationen. Ann. Chem. 522, 75–96 (1936)
Criegee, R.: Organische Osmiumverbindungen. Angew. Chem. 51, 519–530 (1938)
Criegee, R., Marchand, B., Wannowius, H.: Zur Kenntnis der Organischen Osmium-verbindungen. Ann. Chem. 550, 99–133 (1942)
Dreher, K. D., Schulman, J. H., Anderson, O. R., Roels, O. A.: The stability and structure of mixed lipid monolayers and bilayers. I. Properties of lipids and lipoprotein monolayers in OsO4 solutions and the role of cholesterol, retinol and tocopherol in stabilizing lecithin monolayers. J. Ultrastruct. Res. 19, 586–599 (1967)
Fleischer, S., Fleischer, B., Stoeckenius, W.: Fine structure of lipid depleted mitochondria. J. Cell Biol. 32, 193–208 (1967)
Fletcher, J. M., Gardner, W. E., Greenfield, B. F., Holdoway, M. J., Rand, M. H.: Magnetic and other studies of ruthenium dioxide and its hydrate. J. chem. Soc. (A), 653–657 (1968)
Fletcher, J. M., Greenfield, B. F., Hardy, C. J., Scargill, D., Woodhead, J. L.: Ruthenium red. J. chem. Soc. 2000–2006 (1961)
Griffith, W. P.: The chemistry of the rarer platinum metals. New York: Interscience 1967
Hake, T.: Studies on the reactions of OsO4 and KMnO4 with amino acids, peptides, and proteins. Lab. Invest. 14, 1208–1212 (1965)
Hayes, T. L., Lindgren, F. T., Gofman, J. W.: A quantitative determination of the osmium tetroxide lipoprotein interaction. J. Cell Biol. 19, 251–255 (1963)
Highton, T. C., Myers, D. B., Rayns, D. G.: The intercellular spaces of synovial tissue. N. Z. med. J. 67, 315–325 (1968)
Hodgman, C. D.: Handbook of chemistry and physics. Cleveland: Chemical Rubber Publ. Co 1962
Huet, Ch., Herzberg, M.: Effects of enzymes and EDTA on ruthenium red and concanavalin A labelling of the cell surface. J. Ultrastruct. Res. 42, 186–199 (1973)
Kalifat, S. R., Dupuy-Coin, A. M., Delarue, J.: Demonstration ultrastructurale de polysaccharides dont certains acides dans le film de surface de l'alvéole pulmonaire. J. Ultrastruct. Res. 32, 572–589 (1970)
Kavanau, J. L.: Micelle formation and transformation. In: Structure and function in biological membranes, Vol. I, p. 26. San Francisco-London-Amsterdam: Holden-Day Inc. 1965
Kelly, D. E.: Fine structure of desmosomes, hemi-desmosomes, and an adepidermal globular layer in developing newt epidermis. J. Cell Biol. 28, 51–72 (1966)
Klima, J.: Fixierungs- und Einbettungsstudien für die Ultrahistologie. In: Proc. 4th Intern. Conf. Electron Microscopy, Vol. II, p. 58. Berlin-Heidelberg-New York: Springer 1960
Korn, E. D.: Synthesis of bis (methyl-9, 10-dihydroxystearate) osmate from methyl oleate and osmium tetroxide under conditions used for fixation of biological material. Biochim. biophys. Acta (Amst.) 116, 317–325 (1966a)
Korn, E. D.: Modification of oleic acid during fixation of amoabae by osmium tetroxide. Biochim. biophys. Acta (Amst.) 116, 325–335 (1966b)
Korn, E. D.: A chromatographic and spectrophotometric study of the products of the reaction of osmium tetroxide with unsaturated lipids. J. Cell Biol. 34, 627–638 (1967)
Kruyt, H. R.: Colloid science, Vol. I. Amsterdam-Houston-New York-London: Elsevier Publ. Co 1952
Litman, R. B., Barrnett, R. J.: The mechanism of the fixation of tissue components by osmium tetroxide via hydrogen bonding. J. Ultrastruct. Res. 38, 63–86 (1972)
Luft, J. H.: Electron microscopy of cell extraneous coats as revealed by ruthenium red staining. J. Cell Biol. 23, 54A-55A (1964)
Luft, J. H.: Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anat. Rec. 171, 347–368 (1971a)
Luft, J. H.: Ruthenium red and violet. II. Fine structural localization in animals tissues. Anat. Rec. 171, 369–416 (1971b)
Martinez-Palomo, A., Brailovsky, C., Bernhard, W.: Ultrastructural modifications of cell surface and intracellular contacts of some transformed cell strains. Cancer Res. 29, 925–937 (1969)
Napolitano, L., Lebaron, F., Scaletti, J.: Preservation of myelin lamellar structure in the absence of lipid. J. Cell Biol. 34, 817–826 (1967)
Pascal, P.: Nouveau traité de chimie minérale, tome XIX (F. Masson, ed.), p. 173. Paris: F. Masson et Cie. 1958
Reimann, B.: Zur Verwendbarkeit von Rutheniumrot als elektronen-mikroskopisches Kontrastierungsmittel. Mikroskopie 16, 224–226 (1961)
Riemersma, J. C.: Osmium tetroxide fixation of lipids: A possible reaction mechanism. Biochim. biophys. Acta (Amst.) 152, 718–727 (1968)
Riemersma, J. C.: Chemical effects of fixation on biological specimens. In: Some biological techniques in electron microscopy (D. F. Parsons, ed.), p. 85. New York-London: Academic Press 1970
Riemersma, J. C., Booij, H. L.: The reactions of osmium tetroxide with lecithin: Application of staining procedures. J. Histochem. Cytochem. 10, 89–95 (1962)
Rosenfeld, C., Paintrand, M., Choquet, C., Venuat, A. M.: Cyclic variations in the ruthenium red stained coat of cells from a synchronized human lymphoblastoid line. Exp. Cell Res. 79, 465–468 (1973)
Schidlovsky, G.: Contrast in multilayer systems after various fixations. Lab. Invest. 14, 1213–1233 (1965)
Schnepf, E.: Zur Cytologie and Physiologie pflanzlicher Drüsen. 1. Teil. Über den Frangschleim der Insektivoren. Flora (Jena) 153, 1–22 (1963)
Stoeckenius, W., Mahr, S. C.: Studies on the reaction of OsO4 with lipids and related compounds. Lab. Invest. 14, 1196–1207 (1965)
Szubinska, B.: Electron microscopy of the interaction of ruthenium violet with the cell membrane complex of amoeba proteins. J. Cell Biol. 23, 92A (1964)
Szubinska, B., Luft, J. H.: Ruthenium red and violet. III. Fine structure of the plasma membrane and extraneous coats in amoebae. Anat. Rec. 171, 417–442 (1971)
Torpier, G., Montagnier, L.: Corrélation entre la transformation des cellules BHK 21 et l'affinité du rouge de ruthénium pour leur membrane cytoplasmique. Ann. Inst. Pasteur (Lille) 20, 203–210 (1969)
Utsumi, K., Oda, T.: Histochemistry and cytochemistry. In: Proceedings of the 4th International Congress of Histochemistry and Cytochemistry (T. Takeuchi, K. Ogawa, S. Fijita, eds.). Kyoto: Japan Society of Histochemistry and Cytochemistry 1972
Vorbrodt, A., Koprowski, H.: Ruthenium red-stained coat of normal and simian virus 40-transformed cells. J. nat. Cancer Inst. 43, 1241–1248 (1969)
Wigglesworth, V. B.: The use of osmium in the fixation and staining of tissues. Proc. roy. Soc. B 147, 185–199 (1957)
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Blanquet, P.R. Ultrahistochemical study on the ruthenium red surface staining. Histochemistry 47, 63–78 (1976). https://doi.org/10.1007/BF00492555
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DOI: https://doi.org/10.1007/BF00492555