期刊
NATURE PROTOCOLS
卷 14, 期 1, 页码 100-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41596-018-0084-8
关键词
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资金
- University of Leeds (UoL ABSL award)
- Wellcome Trust [108466/Z/15/Z]
- BBSRC [BB/L021250/1]
- European Research Council (ERC) under the European Union's Seventh Framework Programme (FP7/2007-2013) ERC grant [322408]
- MRC [MR/P018491/1]
- BBSRC [BB/L021250/1] Funding Source: UKRI
- MRC [MR/P018491/1] Funding Source: UKRI
The dramatic growth in the use of cryo-electron microscopy (cryo-EM) to generate high-resolution structures of macromolecular complexes has changed the landscape of structural biology. The majority of structures deposited in the Electron Microscopy Data Bank (EMDB) at higher than 4-A resolution were collected on Titan Krios microscopes. Although the pipeline for single-particle data collection is becoming routine, there is much variation in how sessions are set up. Furthermore, when collection is under way, there are a range of approaches for efficiently moving and preprocessing these data. Here, we present a standard operating procedure for single-particle data collection with Thermo Fisher Scientific EPU software, using the two most common direct electron detectors (the Thermo Fisher Scientific Falcon 3 (F3EC) and the Gatan K2), as well as a strategy for structuring these data to enable efficient pre-processing and on-the-fly monitoring of data collection. This protocol takes 3-6 h to set up a typical automated data collection session.
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