Journal
JOURNAL OF STRUCTURAL BIOLOGY
Volume 192, Issue 2, Pages 174-178Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2015.09.003
Keywords
Cryo-EM; Automation; Dose fractionation
Funding
- HHMI
- NSF [DBI-0960271]
- NIH [5R01GM082893, 5R01GM098672, 2P50GM082250, 1S100D020054]
- UCSF Program for Breakthrough Biomedical Research
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Newly developed direct electron detection cameras have a high image output frame rate that enables recording dose fractionated image stacks of frozen hydrated biological samples by electron cryomicroscopy (cryoEM). Such novel image acquisition schemes provide opportunities to analyze ciyoEM data in ways that were previously impossible. The file size of a dose fractionated image stack is 20-60 times larger than that of a single image. Thus, efficient data acquisition and on-the-fly analysis of a large number of dose-fractionated image stacks become a serious challenge to any cryoEM data acquisition system. We have developed a computer-assisted system, named UCSFImage4, for semi-automated cryo-EM image acquisition that implements an asynchronous data acquisition scheme. This facilitates efficient acquisition, on-the-fly motion correction, and CTF analysis of dose fractionated image stacks with a total time of similar to 60 s/exposure. Here we report the technical details and configuration of this system. (C) 2015 Elsevier Inc. All rights reserved.
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