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Nat Commun. 2017 Sep 20;8(1):629. doi: 10.1038/s41467-017-00782-3.

Measuring the effects of particle orientation to improve the efficiency of electron cryomicroscopy.

Author information

1
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
2
Trinity College, University of Cambridge, Cambridge, CB2 1TQ, UK.
3
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK. crusso@mrc-lmb.cam.ac.uk.

Abstract

The orientation distribution of a single-particle electron cryomicroscopy specimen limits the resolution of the reconstructed density map. Here we define a statistical quantity, the efficiency, E od, which characterises the orientation distribution via its corresponding point spread function. The efficiency measures the ability of the distribution to provide uniform information and resolution in all directions of the reconstruction, independent of other factors. This metric allows rapid and rigorous evaluation of specimen preparation methods, assisting structure determination to high resolution with minimal data.A number of parameters influence the resolution of a cryo-EM structure. Here the authors investigate the effects of specimen orientation in single particle cryo-EM and present open-source software for rapidly assessing orientation distributions to improve data collection.

PMID:
28931821
PMCID:
PMC5607000
DOI:
10.1038/s41467-017-00782-3
[Indexed for MEDLINE]
Free PMC Article

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