Sample preparation induced artifacts in cryo-electron tomographs

Pavel Plevka; Anthony J. Battisti; Dennis C. Winkler; Kaspars Tars; Heather A. Holdaway; Carol M. Bator; Michael G. Rossmann

doi:10.1017/S1431927612001298

Sample preparation induced artifacts in cryo-electron tomographs

Pavel Plevka, Anthony J. Battisti, Dennis C. Winkler, Kaspars Tars, Heather A. Holdaway, Carol M. Bator, Michael G. Rossmann

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

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.

Original language	English (US)
Pages (from-to)	1043-1048
Number of pages	6
Journal	Microscopy and Microanalysis
Volume	18
Issue number	5
DOIs	https://doi.org/10.1017/S1431927612001298
State	Published - Oct 2012
Externally published	Yes

Keywords

artifacts
cryo-EM
ice-vacuum interface
microscopy
tomography
virus

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10.1017/S1431927612001298

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title = "Sample preparation induced artifacts in cryo-electron tomographs",

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.",

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T1 - Sample preparation induced artifacts in cryo-electron tomographs

AU - Plevka, Pavel

AU - Battisti, Anthony J.

AU - Winkler, Dennis C.

AU - Tars, Kaspars

AU - Holdaway, Heather A.

AU - Bator, Carol M.

AU - Rossmann, Michael G.

PY - 2012/10

Y1 - 2012/10

N2 - 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.

AB - 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.

KW - artifacts

KW - cryo-EM

KW - ice-vacuum interface

KW - microscopy

KW - tomography

KW - virus

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JO - Microscopy and Microanalysis

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Sample preparation induced artifacts in cryo-electron tomographs

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