Rawson, S, Iadanza, MG, Ranson, NA et al. (1 more author) (2016) Methods to account for movement and flexibility in cryo-EM data processing. Methods, 100. pp. 35-41. ISSN 1046-2023
Abstract
Recent advances in direct electron detectors and improved CMOS cameras have been accompanied by the development of a range of software to take advantage of the data they produce. In particular they allow for the correction of two types of motion in cryo electron microscopy samples: motion correction for movements of the sample particles in the ice, and differential masking to account for heterogeneity caused by flexibility within protein complexes. Here we provide several scripts that allow users to move between RELION and standalone motion correction and centring programs. We then compare the computational cost and improvements in data quality with each program. We also describe our masking procedures to account for conformational flexibility. For the different elements of this study we have used three samples; a high symmetry virus, flexible protein complex (~1 MDa) and a relatively small protein complex (~550 kDa), to benchmark four widely available motion correction packages. Using these as test cases we demonstrate how motion correction and differential masking, as well as an additional particle re-centring protocol can improve final reconstructions when used within the RELION image-processing package.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2016, The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Cryo-EM; RELION; Image processing; Motion correction |
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) |
Funding Information: | Funder Grant number EU - European Union 322408 MRC G1000567/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 26 May 2016 11:21 |
Last Modified: | 11 Apr 2018 13:20 |
Published Version: | http://dx.doi.org/10.1016/j.ymeth.2016.03.011 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.ymeth.2016.03.011 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:97356 |