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Sample Preparation Induced Artifacts in Cryo-Electron Tomographs

Published online by Cambridge University Press:  08 October 2012

Pavel Plevka ,
Anthony J. Battisti ,
Dennis C. Winkler ,
Kaspars Tars ,
Heather A. Holdaway ,
Carol M. Bator  and
Michael G. Rossmann
Pavel Plevka
Affiliation:
Department of Biological Sciences, Purdue University, 240 S. Martin Jischke Drive, West Lafayette, IN 47907-2032, USA
Anthony J. Battisti
Affiliation:
Department of Biological Sciences, Purdue University, 240 S. Martin Jischke Drive, West Lafayette, IN 47907-2032, USA
Dennis C. Winkler
Affiliation:
Laboratory of Structural Biology Research, National Institute for Arthritis, Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892, USA
Kaspars Tars
Affiliation:
Latvian Biomedical Research and Study Centre, Ratsupites Str. 1, Riga, LV 1067, Latvia
Heather A. Holdaway
Affiliation:
Department of Biological Sciences, Purdue University, 240 S. Martin Jischke Drive, West Lafayette, IN 47907-2032, USA
Carol M. Bator
Affiliation:
Department of Biological Sciences, Purdue University, 240 S. Martin Jischke Drive, West Lafayette, IN 47907-2032, USA
Michael G. Rossmann*
Affiliation:
Department of Biological Sciences, Purdue University, 240 S. Martin Jischke Drive, West Lafayette, IN 47907-2032, USA
*
*Corresponding author. E-mail: mr@purdue.edu
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Abstract

We investigated the effects of sample preparation and of the exposure to an electron beam on particles in cryo-electron tomographs. Various virus particles with icosahedral symmetry were examined, allowing a comparison of symmetrically related components that should be identical in structure but might be affected differently by these imaging artifacts. Comparison of tomographic reconstructions with previously determined structures established by an independent method showed that neither freezing nor electron beam exposure produced a significant amount of shrinkage along the z axis (thickness). However, we observed damage to regions of the particles located close to the surface of the vitreous ice.

Keywords

cryo-EMmicroscopyice-vacuum interfaceartifactsvirustomography
Type
Techniques and Equipment Development
Information
Microscopy and Microanalysis , Volume 18 , Issue 5 , October 2012 , pp. 1043 - 1048
Copyright
Copyright © Microscopy Society of America 2012

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Footnotes

†

Present address: Cleveland Center for Membrane and Structural Biology, Case Western Reserve University, Cleveland, OH 44106, USA

𠈁

Present address: Animal Disease Diagnostic Laboratory, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA

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