title: Asynchronous data acquisition and on-the-fly analysis of dose fractionated cryoEM images by UCSFImage.
abstract: 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 cryoEM 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 ∼60s/exposure. Here we report the technical details and configuration of this system.
authors.affiliation: The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94143, United States.
authors.forename: Xueming
authors.initials: X
authors.lastname: Li
authors.affiliation: The Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States.
authors.forename: Shawn
authors.initials: S
authors.lastname: Zheng
authors.affiliation: The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94143, United States; The Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States. Electronic address: agard@msg.ucsf.edu.
authors.forename: David A
authors.initials: DA
authors.lastname: Agard
authors.affiliation: The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94143, United States; The Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States. Electronic address: ycheng@ucsf.edu.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Cheng
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D016247:Information Storage and Retrieval / Q000379:methods; D011506:Proteins / Q000032:analysis*
chemical_list: D011506:Proteins
keywords: Automation; Cryo-EM; Dose fractionation