Montage electron tomography of vitrified specimens

Peck, A., Carter, S. D. , Mai, H., Chen, S., Burt, A. and Jensen, G. J. (2022) Montage electron tomography of vitrified specimens. Journal of Structural Biology, 214(2), 107860. (doi: 10.1016/j.jsb.2022.107860) (PMID:35487464)

Abstract

Cryo-electron tomography provides detailed views of macromolecules in situ. However, imaging a large field of view to provide more cellular context requires reducing magnification during data collection, which in turn restricts the resolution. To circumvent this trade-off between field of view and resolution, we have developed a montage data collection scheme that uniformly distributes the dose throughout the specimen. In this approach, sets of slightly overlapping circular tiles are collected at high magnification and stitched to form a composite projection image at each tilt angle. These montage tilt-series are then reconstructed into massive tomograms with a small pixel size but a large field of view. For proof-of-principle, we applied this method to the thin edge of HeLa cells. Thon rings to better than 10 Å were detected in the montaged tilt-series, and diverse cellular features were observed in the resulting tomograms. These results indicate that the additional dose required by this technique is not prohibitive to performing structural analysis to intermediate resolution across a large field of view. We anticipate that montage tomography will prove particularly useful for lamellae, increase the likelihood of imaging rare cellular events, and facilitate visual proteomics.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Carter, Dr Stephen

Creator Roles

Carter, S. D.: Conceptualization, Methodology, Investigation

• Authors: Peck, A., Carter, S. D., Mai, H., Chen, S., Burt, A., and Jensen, G. J.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Journal of Structural Biology
• Publisher: Elsevier
• ISSN: 1047-8477
• ISSN (Online): 1095-8657
• Published Online: 26 April 2022
• Copyright Holders: Copyright © 2022 Elsevier Inc.
• First Published: First published in Journal of Structural Biology 214(2): 107860
• Publisher Policy: Reproduced in accordance with the publisher copyright policy