期刊
PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY
卷 160, 期 -, 页码 97-103出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.pbiomolbio.2020.05.010
关键词
cryo-ET; cryo-electron tomography; Electron cryotomography; Sub-tomogram averaging; in situ structural biology
资金
- Sir Henry Dale Fellowship - Wellcome Trust [202231/Z/16/Z]
- Sir Henry Dale Fellowship - Royal Society [202231/Z/16/Z]
- Vallee Research Foundation
- Action Medical Research Charity for Children [GN2634]
- Cystic Fibrosis trust
- John Fell Fund
- Medical Research Council graduate studentship
- Wellcome Trust [202231/Z/16/Z] Funding Source: Wellcome Trust
The review discusses new advances in electron cryotomography to address throughput and technical problems, focusing on solutions for sample thinning, improving data collection speed, strategies for target localisation, and advancements in resolution. These advances will significantly decrease the time and effort required for cryo-ET and sub-tomogram averaging.
Electron cryotomography is a rapidly evolving method for imaging macromolecules directly within the native environment of cells and tissues. Combined with sub-tomogram averaging, it allows structural and cell biologists to obtain sub-nanometre resolution structures in situ. However, low throughput in cryo-ET sample preparation and data acquisition, as well as difficulties in target localisation and subtomogram averaging image processing, limit its widespread usability. In this review, we discuss new advances in the field that address these throughput and technical problems. We focus on recent efforts made to resolve issues in sample thinning, improvement in data collection speed at the microscope, strategies for localisation of macromolecules using correlated light and electron microscopy and advancements made to improve resolution in sub-tomogram averaging. These advances will considerably decrease the amount of time and effort required for cryo-ET and sub-tomogram averaging, ushering in a new era of structural biology where in situ macromolecular structure determination will be routine. (C) 2020 The Authors. Published by Elsevier Ltd.
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