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Electron Beam Coater for Reduction of Charging in Ice-Embedded Biological Specimens using Ti88Si12 Alloy

Published online by Cambridge University Press:  21 November 2003

Michael B. Sherman  and
Wah Chiu
Michael B. Sherman
Affiliation:
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030
Wah Chiu
Affiliation:
National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030
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Abstract

Biological macromolecules embedded in vitreous ice are known to suffer from charging while being imaged in an electron transmission cryomicroscope. We developed an electron beam coater that deposits conductive films onto the surface of frozen-hydrated specimens. The conductive films help to dissipate charge during electron irradiation of poorly conductive ice-embedded biological samples. We observed significant reduction in charging of ice-embedded catalase crystals suspended over holes in a holey carbon film after coating them with a 30-Å-thick layer of an amorphous alloy, Ti88Si12. Images of the crystals after coating showed diffraction spots of up to 3 Å resolution.

Keywords

electron cryomicroscopychargingconductive coating
Type
Microscopy Techniques
Information
Microscopy and Microanalysis , Volume 9 , Issue 6 , December 2003 , pp. 566 - 573
Copyright
© 2003 Microscopy Society of America

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References

REFERENCES

Avila-Sakar, A.J. & Chiu, W. (1996). Visualization of beta-sheets and side-chain clusters in two-dimensional periodic arrays of streptavidin on phospholipid monolayers by electron crystallography. Biophys J 70, 57–68.CrossRefGoogle Scholar
Böttcher, B. (1995). Electron cryo-microscopy of graphite in amorphous ice. Ultramicroscopy 58, 417–424.CrossRefGoogle Scholar
Brink, J. & Chiu, W. (1994). Applications of a slow-scan CCD camera in protein electron crystallography. J Struct Biol 113, 23–34.CrossRefGoogle Scholar
Brink, J., Gross, H., Tittmann, P., Sherman, M.B., & Chiu, W. (1998a). Reduction of charging in protein electron cryomicroscopy. J Microsc 191, 67–73.Google Scholar
Brink, J., Sherman, M.B., Berriman, J., & Chiu, W. (1998b). Evaluation of charging on macromolecules in electron cryomicroscopy. Ultramicroscopy 72, 41–52.Google Scholar
Dubochet, J., Adrian, M., Chang, J.J., Homo, J.C., Lepault, J., McDowall, A.W., & Schultz, P. (1988). Cryo-electron microscopy of vitrified specimens. Q Rev Biophys 21, 129–228.CrossRefGoogle Scholar
Fukami, A. & Adachi, K. (1965). A new method of preparation of a self-perforated micro plastic grid and its application. J Elect Microsc 14, 112–118.Google Scholar
Henderson, R. (1992). Image contrast in high-resolution electron microscopy of biological macromolecules: TMV in ice. Ultramicroscopy 46, 1–18.Google Scholar
Jakubowski, U., Baumeister, W., & Glaeser, R.M. (1989). Evaporated carbon stabilizes thin, frozen-hydrated specimens. Ultramicroscopy 31, 351–356.CrossRefGoogle Scholar
Miyazawa, A., Fujiyoshi, Y., Stowell, M., & Unwin, N. (1999). Nicotinic acetylcholine receptor at 4.6 A resolution: Transverse tunnels in the channel wall. J Mol Biol 288, 765–786.Google Scholar
Pawley, J. (1984). Low voltage scanning electron microscopy. J Microsc 136, 45–68.CrossRefGoogle Scholar
Rader, R.S. & Lamvik, M.K. (1992). High-conductivity amorphous TiSi substrates for low temperature electron microscopy. J Microsc 168, 71–77.CrossRefGoogle Scholar
Sherman, M.B., Brink, J., & Chiu, W. (1996). Performance of a slow-scan CCD camera for macromolecular imaging in a 400 kV electron cryomicroscope. Micron 27, 129–139.CrossRefGoogle Scholar
Warrington, D.H. (1966). A simple charge neutralizer for the electron microscope. J Sci Instrum 43, 77–78.CrossRefGoogle Scholar
Williams, D.B. & Carter, C.B. (1996). Transmission Electron Microscopy: Basics I. New York: Plenum Press.CrossRef
Wrigley, N.G. (1968). The lattice spacing of crystalline catalase as an internal standard of length in electron microscopy. J Ultrastruct Res 24, 454–464.CrossRefGoogle Scholar