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  Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution subtomogram averaging

Hagen, W. J. H., Wan, W., & Briggs, J. A. G. (2017). Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution subtomogram averaging. Journal of Structural Biology, 197(2), 191-198. doi:10.1016/j.jsb.2016.06.007.

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 Creators:
Hagen, W. J. H., Author
Wan, W., Author
Briggs, John A. G.1, Author           
Affiliations:
1European Molecular Biology Laboratory, External Organizations, ou_3346677              

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Free keywords: Electron tomography Tilt-scheme Subtomogram averaging Cryo-electron microscopy electron cryotomography alignment Biochemistry & Molecular Biology Biophysics Cell Biology
 Abstract: Cryo-electron tomography (cryoET) allows 3D structural information to be obtained from cells and other biological samples in their close-to-native state. In combination with subtomogram averaging, detailed structures of repeating features can be resolved. CryoET data is collected as a series of images of the sample from different tilt angles; this is performed by physically rotating the sample in the microscope between each image. The angles at which the images are collected, and the order in which they are collected, together are called the tilt-scheme. Here we describe a "dose-symmetric tilt-scheme" that begins at low tilt and then alternates between increasingly positive and negative tilts. This tilt-scheme maximizes the amount of high-resolution information maintained in the tomogram for subsequent subtomogram averaging, and may also be advantageous for other applications. We describe implementation of the tilt-scheme in combination with further data-collection refinements including setting thresholds on acceptable drift and improving focus accuracy. Requirements for microscope set-up are introduced, and a macro is provided which automates the application of the tilt-scheme within SerialEM. (C) 2016 The Authors. Published by Elsevier Inc.

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Language(s): eng - English
 Dates: 2017
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: Other: WOS:000393268200014
DOI: 10.1016/j.jsb.2016.06.007
ISSN: 1047-8477
 Degree: -

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Title: Journal of Structural Biology
  Alternative Title : J. Struct. Biol.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 197 (2) Sequence Number: - Start / End Page: 191 - 198 Identifier: -