A complete data processing workflow for cryo-ET and subtomogram averaging

Electron cryotomography is currently the only method capable of visualizing cells in three dimensions at nanometer resolutions. While modern instruments produce massive amounts of tomography data containing extremely rich structural information, data processing is very labor intensive and the result...

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Vydáno v:Nature methods Ročník 16; číslo 11; s. 1161 - 1168
Hlavní autoři: Chen, Muyuan, Bell, James M, Shi, Xiaodong, Sun, Stella Y, Wang, Zhao, Ludtke, Steven J
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States Nature Publishing Group 01.11.2019
Témata:
Alignment
Cryoelectron Microscopy - methods
Data processing
Electronic Data Processing - methods
Human bias
Image Processing, Computer-Assisted - methods
Information processing
Macromolecules
Tomography
Transfer functions
Workflow
ISSN:1548-7091, 1548-7105, 1548-7105
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Shrnutí:Electron cryotomography is currently the only method capable of visualizing cells in three dimensions at nanometer resolutions. While modern instruments produce massive amounts of tomography data containing extremely rich structural information, data processing is very labor intensive and the results are often limited by the skills of the personnel rather than the data. We present an integrated workflow that covers the entire tomography data processing pipeline, from automated tilt series alignment to subnanometer resolution subtomogram averaging. Resolution enhancement is made possible through the use of per-particle per-tilt contrast transfer function correction and alignment. The workflow greatly reduces human bias, increases throughput and more closely approaches data-limited resolution for subtomogram averaging in both purified macromolecules and cells.
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ISSN:1548-7091
1548-7105
1548-7105
DOI:10.1038/s41592-019-0591-8