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Rapid Tilt-Series Acquisition for Electron Cryotomography

Chreifi, Georges and Chen, Songye and Metskas, Lauren Ann and Kaplan, Mohammed and Jensen, Grant J. (2019) Rapid Tilt-Series Acquisition for Electron Cryotomography. Journal of Structural Biology, 205 (2). pp. 163-169. ISSN 1047-8477. PMCID PMC6389375. https://resolver.caltech.edu/CaltechAUTHORS:20181107-100852395

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Abstract

Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a tilt-series in seconds, but with loss of details finer than ∼4 nm. We also demonstrate a fast-incremental method that can record a tilt-series several-fold faster than current methods and with similar resolution. We characterize the utility of both methods in real biological electron cryotomography workflows. We identify opportunities for further improvements in hardware and software and speculate on the impact such advances could have on structural biology.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.jsb.2018.12.008DOIArticle
https://doi.org/10.1101/454587DOIDiscussion Paper
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389375/PubMed CentralArticle
ORCID:
AuthorORCID
Chen, Songye0000-0001-5407-5049
Metskas, Lauren Ann0000-0002-8073-6960
Kaplan, Mohammed0000-0002-0759-0459
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2019 Elsevier Inc. Received 28 October 2018, Revised 18 December 2018, Accepted 20 December 2018, Available online 11 January 2019. We thank David N. Mastronarde for helpful suggestions and for providing comments on this manuscript. 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. M.K. is supported by a postdoctoral Rubicon fellowship from De Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO).
Funders:
Funding AgencyGrant Number
NIHGM122588
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)UNSPECIFIED
Issue or Number:2
PubMed Central ID:PMC6389375
Record Number:CaltechAUTHORS:20181107-100852395
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20181107-100852395
Official Citation:Georges Chreifi, Songye Chen, Lauren Ann Metskas, Mohammed Kaplan, Grant J. Jensen, Rapid tilt-series acquisition for electron cryotomography, Journal of Structural Biology, Volume 205, Issue 2, 2019, Pages 163-169, ISSN 1047-8477, https://doi.org/10.1016/j.jsb.2018.12.008. (http://www.sciencedirect.com/science/article/pii/S1047847719300048)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90701
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:07 Nov 2018 19:26
Last Modified:03 Oct 2019 20:27

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