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Rapid tilt-series method for cryo-electron tomography: Characterizing stage behavior during FISE acquisition

Chreifi, Georges and Chen, Songye and Jensen, Grant J. (2021) Rapid tilt-series method for cryo-electron tomography: Characterizing stage behavior during FISE acquisition. Journal of Structural Biology, 213 (2). Art. No. 107716. ISSN 1047-8477. doi:10.1016/j.jsb.2021.107716. https://resolver.caltech.edu/CaltechAUTHORS:20200522-113331410

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Abstract

We and others recently developed rapid tilt-series acquisition methods for cryo-electron tomography on a Titan Krios G3i equipped with a single axis holder and a K-series direct electron detector and showed that one of these, the fast-incremental single exposure (FISE) method, significantly accelerates tilt-series acquisition when compared to traditional methods while preserving the quality of the images. Here, we characterize the behavior of our single axis holder in detail during a FISE experiment to optimally balance data quality with speed. We explain our methodology in detail so others can characterize their own stages, and conclude with recommendations for projects with different resolution goals.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.jsb.2021.107716DOIArticle
https://doi.org/10.1101/2020.05.19.104828DOIDiscussion Paper
ORCID:
AuthorORCID
Chreifi, Georges0000-0003-4194-1694
Chen, Songye0000-0001-5407-5049
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2021 Published by Elsevier Inc. Received 22 May 2020, Revised 2 March 2021, Accepted 5 March 2021, Available online 11 March 2021. This work was supported by NIH grant GM122588 (to G.J.J.). Electron cryomicroscopy was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. CRediT authorship contribution statement: Georges Chreifi: Conceptualization, Methodology, Software, Investigation, Writing - review & editing. Songye Chen: Supervision, Methodology. Grant J. Jensen: Conceptualization, Methodology, Writing - review & editing. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Funders:
Funding AgencyGrant Number
NIHGM122588
Subject Keywords:Cryo-ET; Cryo-EM; Tomography; Tilt series; K3; FISE
Issue or Number:2
DOI:10.1016/j.jsb.2021.107716
Record Number:CaltechAUTHORS:20200522-113331410
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200522-113331410
Official Citation:Georges Chreifi, Songye Chen, Grant J. Jensen, Rapid tilt-series method for cryo-electron tomography: Characterizing stage behavior during FISE acquisition, Journal of Structural Biology, Volume 213, Issue 2, 2021, 107716, ISSN 1047-8477, https://doi.org/10.1016/j.jsb.2021.107716. (https://www.sciencedirect.com/science/article/pii/S1047847721000216)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:103403
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:22 May 2020 18:47
Last Modified:29 Mar 2021 21:15

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