Summary
Fine grain development for electron microscopic radioautography was investigated with two types of radioactive specimens: sections of tritiated methacrylate, which provide a homogeneously labeled source for quantitative evaluation of the radioautographic reaction, and sections of 125I-labeled thyroid. Radioautographs were prepared with Ilford L4, Sakura NR-H2, Agfa-Gevaert NUC 307 or Kodak NTE emulsions. The radioautographs were developed with one of several “solution physical” development procedures (Agfa-Gevaert, phenidone-ascorbic acid, p-phenylenediamine developers) or with arrested “direct” developments (D-19b, Elon-ascorbic acid developers). By arresting each development at an early stage of the reaction and at progressively longer time intervals, it was possible to examine the sequence of shapes in the growth of developed silver deposits for each emulsion-development combination. Thus, conditions which resulted in the development of small, round, compact silver deposits were defined for each emulsion.
These developments were used in conjuction with gold latensification, a treatment which increases the sensitivity of the emulsions and thus compensates for the lowered sensitivity of fine grain development procedures.
The location of the silver deposits in relation to the silver bromide crystals from which they derive was investigated. The emulsion gelatin surrounding the crystals was stained whereas the spaces, which remained after the crystals were dissolved in the photographic fixer, appeared transparent. This analysis permitted the selection of development procedures in which the single or multiple round silver deposits originating from a single crystal will remain within or on the boundary of this crystal. By this method, quantitation of radioautographic reactions composed of small, round silver deposits was studied by using the uniformly labeled 3H-methacrylate sections as a standard source of radiation. The conditions under which grain counting is feasible are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agfa-Gevaert data sheet, emulsions for nuclear physics and autoradiography
Caro, L. G., Van Tubergen, R. P.: High resolution autoradiography. I. Methods. J. Cell Biol. 15, 173–188 (1962)
Heremans, H. A.: Combination of electron microscope and autoradiography using a fine grain nuclear emulsion. Histochemie 25, 123–129 (1971)
Hornsby, K. M.: Basic Photographic Chemistry. The Fountain Press in conjunction with the British Journal of Photography 1958
James, T. H.: The site of reaction in direct photographic development. II. Kinetics of development initiated by gold nuclei. J. Colloid Sci. 3, 447–455 (1948)
James, T. H.: Some views on the mechanism of development. J. Phys. Chem. 66, 2416–2422 (1962)
James, T. H., Vanselow, W., Quirk, R. F.: Gold and mercury latensification and hypersensitization for direct and physical development. PSA Journal 14, 349–353 (1948)
Kopriwa, B. M.: A semiautomatic instrument for the radioautographic coating technique. J. Histochem. Cytochem. 14, 923–928 (1967a)
Kopriwa, B. M.: The influence of development on the number and appearance of silver grains in electron microscopic radioautography. J. Histochem. Cytochem. 15, 501–515 (1967b)
Kopriwa, B. M.: A reliable, standardized method for ultrastructural electron microscopic radioautography. Histochemie 37, 1–17 (1973)
Kopriwa, B. M., Nadler, N. J.: Comparison of Kodak D-19b and Agfa-Gevaert solution physical developments in radioautographs of thyroid follicles after injection of 125I to the rat. (In preparation)
Lettré, H., Paweletz, N.: Probleme der elektronenmikroskopischen Autoradiographie. Naturwissenschaften 53, 268–271 (1966)
Mees, C. E. K., James, T. H.: The theory of the photographic process, third ed. New York-London: The Macmillan Company 1966
Nicholae, M.: Developpement discriminatoire de traces nucléaires pour la microscopie électronique des émulsions ionographiques. Sci. Indust. phot. 37, 161–165 (1966)
Rechenmann, R. V., Wittendorp, E.: High efficiency development procedures for nuclear emulsions. J. Microscopy 96, 227–244 (1972)
Salpeter, M. M., Bachmann, L.: Autoradiography with the electron microscope. J. Cell Biol. 22, 469–477 (1964)
Uchida, K., Mizuhira, V.: Electron microscope autoradiography with special reference to the problem of resolution. Arch. histol. jap. 31, 291–320 (1970)
Weber, H.: Einige Erfahrungen bei der Anwendung der elektronenmikroskopischen Autoradiographie. Acta biol. med. germ. 22, 159–167 (1969)
Wisse, E., Tates, A. D.: A gold latensification-Elon ascorbic acid developer for Ilford L4 emulsion. Fourth European Regional Conference on Electron Microscopy, Rome, p. 465–466 (1968)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kopriwa, B.M. A comparsion of various procedures for fine grain development in electron microscopic radioautography. Histochemistry 44, 201–224 (1975). https://doi.org/10.1007/BF00491492
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00491492