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. 1962 Nov 1;15(2):173–188. doi: 10.1083/jcb.15.2.173

HIGH-RESOLUTION AUTORADIOGRAPHY

I. Methods

Lucien G Caro 1, Robert P van Tubergen 1
PMCID: PMC2106152  PMID: 14018772

Abstract

Methods used in obtaining high resolution in autoradiography, with special emphasis on the technique of electron microscopic autoradiography, are described, together with control experiments designed to establish the optimum conditions or procedures. On the basis of these experiments the emulsion selected was Ilford L-4, with a crystal size slightly larger than 0.1 micron. It is applied to the specimen in the form of a gelled film consisting of a monolayer of silver halide crystals. Background, when present, can be eradicated by a simple method. The preparations can be stored, in presence of a drying agent, at room temperature or in a refrigerator. Photographic development is done in Microdol, or in a special fine grain "physical" developer. For examination in the electron microscope the sections are stained with uranyl or lead stains. These methods give a good localization of the label, at the subcellular level, and good reproducibility in relative grain counts.

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Selected References

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  1. CARO L. G. Electron microscopic radioautography of thin sections: the Golgi zone as a site of protein concentration in pancreatic acinar cells. J Biophys Biochem Cytol. 1961 May;10:37–45. doi: 10.1083/jcb.10.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CARO L. G., FORRO F., Jr Localization of macromolecules in Escherichia coli. II. RNA and its site of synthesis. J Biophys Biochem Cytol. 1961 Mar;9:555–565. doi: 10.1083/jcb.9.3.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CARO L. G. High-resolution autoradiogaphy. II. The problem of resolution. J Cell Biol. 1962 Nov;15:189–199. doi: 10.1083/jcb.15.2.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CARO L. G., PALADE G. E. [The role of the Golgi apparatus in the secretory process. Autoradiographic study]. C R Seances Soc Biol Fil. 1961;155:1750–1762. [PubMed] [Google Scholar]
  5. Comer J. J., Skipper S. J. Nuclear Emulsions for Electron Microscopy. Science. 1954 Apr 2;119(3092):441–442. doi: 10.1126/science.119.3092.441. [DOI] [PubMed] [Google Scholar]
  6. GIBBONS I. R., GRIMSTONE A. V. On flagellar structure in certain flagellates. J Biophys Biochem Cytol. 1960 Jul;7:697–716. doi: 10.1083/jcb.7.4.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HAMPTON J. C., QUASTLER H. Combined autoradiography and electron microscopy of thin sections of intestinal epithelial cells of the mouse labeled with H3-thymidine. J Biophys Biochem Cytol. 1961 May;10:140–144. doi: 10.1083/jcb.10.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HARFORD C. G., HAMLIN A. Electron microscopic autoradiography in a magnetic field. Nature. 1961 Feb 11;189:505–506. doi: 10.1038/189505a0. [DOI] [PubMed] [Google Scholar]
  9. LEVI H. A discussion of recent advance towards quantitative autoradiography. Exp Cell Res. 1957;13(Suppl 4):207–221. [PubMed] [Google Scholar]
  10. MESSIER B., LEBLOND C. P. Preparation of coated radioautographs by dipping sections in fluid emulsion. Proc Soc Exp Biol Med. 1957 Oct;96(1):7–10. doi: 10.3181/00379727-96-23380. [DOI] [PubMed] [Google Scholar]
  11. RAY R. C., STEVENS G. W. W. Reciprocity failure and latent-image fading in autoradiography. Br J Radiol. 1953 Jul;26(307):362–367. doi: 10.1259/0007-1285-26-307-362. [DOI] [PubMed] [Google Scholar]
  12. REVEL J. P., HAY E. D. Autoradiographic localization of DNA synthesis in a specific ultrastructural, component of the interphase nucleus. Exp Cell Res. 1961 Nov;25:474–480. doi: 10.1016/0014-4827(61)90301-9. [DOI] [PubMed] [Google Scholar]
  13. SATIR P. G., PEACHEY L. D. Thin sections. II. A simple method for reducing compression artifacts. J Biophys Biochem Cytol. 1958 May 25;4(3):345–348. doi: 10.1083/jcb.4.3.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. VAN TUBERGEN R. P., SETLOW R. B. Quantitative radioautographic studies on exponentially growing cultures of Escherichia coli. The distribution of parental DNA, RNA, protein, and cell wall among progeny cells. Biophys J. 1961 Sep;1:589–625. doi: 10.1016/s0006-3495(61)86911-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. VAN TUBERGEN R. P. The use of radioautography and electron microscopy for the localization of tritium label in bacteria. J Biophys Biochem Cytol. 1961 Jan;9:219–222. doi: 10.1083/jcb.9.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]

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