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Electron Digital Imaging toward High-Resolution Structure Analysis of Biological Macromolecules

Published online by Cambridge University Press:  04 July 2008

Saori Maki-Yonekura  and
Koji Yonekura
Saori Maki-Yonekura
Affiliation:
The W. M. Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, 1700 4th Street, San Francisco, CA 94158-2532, USA
Koji Yonekura*
Affiliation:
The W. M. Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, 1700 4th Street, San Francisco, CA 94158-2532, USA
*
Corresponding author. E-mail: [email protected]
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Abstract

Digital imaging has been applied to structure analysis of biological macromolecules in combination with electron energy filtering. Energy filtering can improve the image contrast of frozen-hydrated specimens, but needs a high-sensitivity imaging device instead of photographic film, because of a decrease in electrons after filtration. Here, a lens-coupled slow-scan charge-coupled device (SSCCD) camera with a post-column-type energy filter were examined to image bacterial flagellar filaments embedded in ice. We first measured the modulation transfer function of this camera and showed the remarkable improvement, compared to other fiber-coupled SSCCD cameras. The 3D structure calculated at ∼7-Å resolution clearly resolves α-helices. Furthermore, filtered datasets recorded on the SSCCD camera with liquid-nitrogen and liquid-helium cooling were compared with the previous unfiltered one on film with liquid-helium cooling. This report describes the suitability of digital imaging with energy filtering for higher-resolution structure studies from its practical application.

Keywords

SSCCDMTFliquid-helium coolingcryoelectron microscopyflagellar filament
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 14 , Issue 4 , August 2008 , pp. 362 - 369
Copyright
Copyright © Microscopy Society of America 2008

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