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Published February 1, 2019 | Accepted Version + Submitted
Journal Article Open

Rapid Tilt-Series Acquisition for Electron Cryotomography

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.

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).

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Accepted Version - nihms-1519590.pdf

Submitted - 454587.full.pdf

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August 19, 2023
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October 20, 2023