Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Jan 1;72(1):161–173. doi: 10.1083/jcb.72.1.161

Resolution in electron microscope autoradiography. III. Iodine-125, the effect of heavy metal staining, and a reassessment of critical parameters

PMCID: PMC2110981  PMID: 63463

Abstract

Resolution for 125I-labeled specimens under electron microscope (EM) autoradiographic conditions was assessed experimentally. With this isotope the size of the silver halide crystal was the most important resolution-limiting factor. Heavy metal staining such as is routinely used in preparing animal tissues for EM autoradiography produced an improvement in resolution of approximately 15-20%. For a 500-1,000-A biological tissue section fixed with OsO4 and stained with uranyl acetate, we obtained resolution (half distance, HD) values of approximately 800 +/- 120 A using Ilford L4 emulsion and 500 +/- 70 A using a Kodak NTE-type emulsion. General aspects of resolution-limiting factors and comparison with 3H and 14C values are discussed.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BACHMANN L., SALPETER M. M. AUTORADIOGRAPHY WITH THE ELECTRON MICROSCOPE; A QUANTITATIVE EVALUATION. Lab Invest. 1965 Jun;14:1041–1053. [PubMed] [Google Scholar]
  2. Blackett N. M., Parry D. M. A new method for analyzing electron microscope autoradiographs using hypothetical grain distributions. J Cell Biol. 1973 Apr;57(1):9–15. doi: 10.1083/jcb.57.1.9. [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., VAN TUBERGEN R. P., KOLB J. A. High-resolution autoradiography. I. Methods. J Cell Biol. 1962 Nov;15:173–188. doi: 10.1083/jcb.15.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fertuck H. C., Salpeter M. M. Quantitation of junctional and extrajunctional acetylcholine receptors by electron microscope autoradiography after 125I-alpha-bungarotoxin binding at mouse neuromuscular junctions. J Cell Biol. 1976 Apr;69(1):144–158. doi: 10.1083/jcb.69.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fertuck H. C., Salpeter M. M. Sensitivity in electron microscope autoradiography for 125I. J Histochem Cytochem. 1974 Feb;22(2):80–87. doi: 10.1177/22.2.80. [DOI] [PubMed] [Google Scholar]
  7. Haworth R. A., Chapman J. A. 125-I in electron microscope autoradiography. Experientia. 1975 Feb 15;31(2):258–260. doi: 10.1007/BF01990740. [DOI] [PubMed] [Google Scholar]
  8. Locke M., Krishnan N., McMahon J. T. A routine method for obtaining high contrast without staining sections. J Cell Biol. 1971 Aug;50(2):540–544. doi: 10.1083/jcb.50.2.540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. MOSES M. J. APPLICATION OF AUTORADIOGRAPHY TO ELECTRON MICROSCOPY. J Histochem Cytochem. 1964 Feb;12:115–130. doi: 10.1177/12.2.115. [DOI] [PubMed] [Google Scholar]
  10. MYERS W. G., VANDERLEEDEN J. C. Radioiodine-125. J Nucl Med. 1960 Jul;1:149–164. [PubMed] [Google Scholar]
  11. Parry D. M., Blackett N. M. Electron microscope autoradiography of erythroid cells using radioactive iron. J Cell Biol. 1973 Apr;57(1):16–26. doi: 10.1083/jcb.57.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. SALPETER M. M., BACHMANN L. AUTORADIOGRAPHY WITH THE ELECTRON MICROSCOPE. A PROCEDURE FOR IMPROVING RESOLUTION, SENSITIVITY, AND CONTRAST. J Cell Biol. 1964 Aug;22:469–477. doi: 10.1083/jcb.22.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Salpeter M. M., Budd G. C., Mattimoe S. Resolution in autoradiography using semithin sections. J Histochem Cytochem. 1974 Apr;22(4):217–222. doi: 10.1177/22.4.217. [DOI] [PubMed] [Google Scholar]
  14. Salpeter M. M., Salpeter E. E. Resolution in electron microscope radioautography. II. Carbon 14. J Cell Biol. 1971 Aug;50(2):324–332. doi: 10.1083/jcb.50.2.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Salpeter M. M. Sensitivity in electron microscope autoradiography. II. Effect of heavy metal staining. J Histochem Cytochem. 1973 Jul;21(7):623–627. doi: 10.1177/21.7.623. [DOI] [PubMed] [Google Scholar]
  16. Salpeter M. M., Szabo M. Sensitivity in electron microscope autoradiography. I. The effect of radiation dose. J Histochem Cytochem. 1972 Jun;20(6):425–434. doi: 10.1177/20.6.425. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES