Summary
The effect of various steps in the preparation of muscle and blood cells for electron microscopic X-ray microanalysis, was studied with reference to calcium, zinc and copper levels.
It was found that these elements are readily detectable in material fixed in glutaraldehyde and embedded directly in Epon 812. Alcohol dehydration, buffer rinses and post-osmication caused marked depletion of all elements examined.
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References
Adelstem, S. J., Vallee, B. L.: Copper. In: Mineral metabolism, vol. II, part B, p. 371, ed. by Comar, C. L. and Bronnes, F. (1962)
Agostini, B., Hasselbach, N.: Electron cytochemistry of calcium uptake: the fragmented sarcoplasmic reticulum. Histochemie 28, 55–67 (1971)
Appleton, T. C.: Dry ultrathin frozen sections for electron microscopy: The cryostat approach. Micron 3, 101 (1971)
Chvapil, M., Elias, S. L., Ryan, J. N., Zukoski, C. F.: Pathophysiology of zinc, p. 105–125. Neurobiology of the trace metals zinc and copper, ed. by Carl C. Pfeiffer. Neurobiol. Suppl. 1, (1972)
Coleman, J. R., Terpka, A. R.: Electron probe analysis of the calcium distribution in cells of the embryonic chick chorioallarbic membrane. J. Histochem. Cytochem. 20, 401 (1972)
Coleman, J. R., Terepka, A. R.: Preparatory methods for electron probe analysis, chap. 8, p. 159–207. In: Principle techniques of electron microscopy, vol. 4, ed. by M. A. Hayat. New York: Van Nostrand-Reinhold Co. 1974
Delves, H. T., Alexander, F. W., Lay, H.: Copper and zinc concentration in the plasma of leukemic children. Brit. J. Haemat. 24, 525–531 (1973)
Dubowitz, V.: Pers. communication (1975)
Garfield, R. E., Henderson, R. M., Daniel, E. E.: Evaluation of the pyroantimonate technique for localization of tissue sodium. Tissue and Cell 4, 515–589 (1972)
Hall, T. A.: The microprobe assay of chemical elements in physical techniques in biological research. Ed. by G. Oster, p. 157, vol. 1A, 2nd ed. New York: Academic Press 1971
Hall, T. A., Rockert, H. O. E., Saunders, R. L.: X-ray microscopy in clinical and experimental medicine. Springfield, Illinois: Charles C. Thomas 1972
Oschman, J. L., Wall, B. J.: Calcium binding to intestinal membranes. J. Cell Biol. 55, 58–73 (1972)
Pease, D. C.: Polar embedments which retain lipids in sections. Proc. 8th Internat. Congr. on Electron Microscopy, Canberra, vol. II, p. 126 (1974)
Pentilla, A., Kalimo, H., Trump, B.: Influence of glutaraldehyde and/or osmium tetroxide on cell volume, ion content, mechanical stability and membrane permeability of Erlich ascites tumor cell. J. Cell Biol. 63, 197–214 (1974)
Rasmussen, K. E.: Fixation in aldehyde. A study on the influence of the fixative, buffer and osmolarity upon the fixation of the rat retina. J. Ultrastruct. Res. 46, 87–102 (1974)
Winegrad, S.: The intracellular site of calcium activation of contraction in frog skeletal muscle. J. gen. Physiol. 55, 77 (1970)
Yarom, R., Chandler, J. A.: Electron probe analysis of skeletal muscle. J. Histochem. Cytochem. 22, 147–154 (1974)
Yarom, R., Peters, P. D., Hall, T. A.: Effect of glutaraldehyde and urea embedding on intracellular ionic elements. J. Ultrastruct. Res. 49, 405–418 (1974)
Yarom, R., Peters, P. D., Hall, T. A.: Calcium in myonuclei, electron microprobe analysis. Experimentia (Basel) 31, 154–157 (1975)
Yarom, R., Peters, P. D., Scripps, M., Rogel, S.: Effect of specimen preparation on intracellular Myocardial calcium. Histochemistry 38, 143–153 (1974)
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Yarom, R., Maunder, C., Scripps, M. et al. A simplified method of specimen preparation for X-ray microanalysis of muscle and blood cells. Histochemistry 45, 49–59 (1975). https://doi.org/10.1007/BF00508050
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DOI: https://doi.org/10.1007/BF00508050