Drulyte, I orcid.org/0000-0003-1117-7699, Johnson, RM, Hesketh, EL et al. (6 more authors) (2018) Approaches to altering particle distributions in cryo-electron microscopy sample preparation. Acta Crystallographica Section D: Structural Biology, D74 (Pt 6). pp. 560-571. ISSN 2059-7983
Abstract
Cryo-electron microscopy (cryo-EM) can now be used to determine high-resolution structural information on a diverse range of biological specimens. Recent advances have been driven primarily by developments in microscopes and detectors, and through advances in image-processing software. However, for many single-particle cryo-EM projects, major bottlenecks currently remain at the sample-preparation stage; obtaining cryo-EM grids of sufficient quality for high-resolution single-particle analysis can require the careful optimization of many variables. Common hurdles to overcome include problems associated with the sample itself (buffer components, labile complexes), sample distribution (obtaining the correct concentration, affinity for the support film), preferred orientation, and poor reproducibility of the grid-making process within and between batches. This review outlines a number of methodologies used within the electron-microscopy community to address these challenges, providing a range of approaches which may aid in obtaining optimal grids for high-resolution data collection.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence [https://creativecommons.org/licenses/by/2.0/uk/legalcode], which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
Keywords: | cryo-electron microscopy; single-particle sample preparation; single-particle analysis. |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Animal Cell Biology (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cryo EM, Image Processing (Leeds) |
Funding Information: | Funder Grant number Wellcome Trust 109158/B/15/A |
Depositing User: | Symplectic Publications |
Date Deposited: | 11 May 2018 10:10 |
Last Modified: | 25 Jun 2023 21:20 |
Status: | Published |
Publisher: | International Union of Crystallography |
Identification Number: | 10.1107/S2059798318006496 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:130715 |