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Use Tungsten Shadowing to Study Biological Macromolecules

Published online by Cambridge University Press:  02 July 2020

Sergey N. Ryazantsev
Sergey N. Ryazantsev*
Affiliation:
Department of Biological Chemistry, School of Medicine, University of California, Los Angeles, Box 951737, Los Angeles, California , 90095-1737, USA
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Abstract

Shadowing is widely used for biological macromolecules and their complexes structure determination . It has been shown that tungsten shadowing produces fine granularity and good structural detail resolution. We have developed a working prototype of an electron beam evaporator (“electron gun“) based on the principle described by V. Vasiliev. The electron gun has characteristics that allow us to deposit high quality films of carbon or even pure tungsten. in addition we have constructed a prototype sample handling apparatus for freeze drying and shadowing. The prototype allows tilting of +/- 90° in 2° increments, rotation (if desired) at 6 rpm, controlled heating at a rate of 1-2° per min, and cooling using a liquid nitrogen trap attached to the device. The system is vacuumed by 480 1/sec maglev turbomolecular pump with scroll pump at the first stage. It produces oil-free vacuum in 2-5*10-7 torr range, which we believe, is sufficient for good quality shadowing.

Type
Recent Techniques for the Fixation and Staining of Biological Samples (Organized by M. Sanders and K. McDonald)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 1206 - 1207
Copyright
Copyright © Microscopy Society of America 2001

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References

References:

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7. This project has been supported by NSF grant 9819814 and NIH grant GM 38 509. DNA samples were kindly prepared by Stacy Merickel and Reid Johnson, UCLA. Human myeloma IgGl samples were kindly prepared by V. Abramov and V. Zav'yalov, Institute of Immunology, Ljubuchany, Russia.Google Scholar