title: Maximizing the potential of electron cryomicroscopy data collected using direct detectors.
abstract: Single-particle electron cryomicroscopy is undergoing a technical revolution due to the recent developments of direct detectors. These new recording devices detect electrons directly (i.e. without conversion into light) and feature significantly improved detective quantum efficiencies and readout rates as compared to photographic films or CCDs. We evaluated here the potential of one such detector (Gatan K2 Summit) to enable the achievement of near-atomic resolution reconstructions of biological specimens when coupled to a widely used, mid-range transmission electron microscope (FEI TF20 Twin). Compensating for beam-induced motion and stage drift provided a 4.4Å resolution map of Sulfolobus turreted icosahedral virus (STIV), which we used as a test particle in this study. Several motion correction and dose fractionation procedures were explored and we describe their influence on the resolution of the final reconstruction. We also compared the quality of this data to that collected with a FEI Titan Krios microscope equipped with a Falcon I direct detector, which provides a benchmark for data collected using a high-end electron microscope.
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.|National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: David
authors.initials: D
authors.lastname: Veesler
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.|National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Melody G
authors.initials: MG
authors.lastname: Campbell
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.|National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Anchi
authors.initials: A
authors.lastname: Cheng
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Chi-Yu
authors.initials: CY
authors.lastname: Fu
authors.affiliation: Department of Computer Science and Engineering, University of California, San Diego, CA 92093.
authors.forename: Zachary
authors.initials: Z
authors.lastname: Murez
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: John E
authors.initials: JE
authors.lastname: Johnson
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.|National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Clinton S
authors.initials: CS
authors.lastname: Potter
authors.affiliation: Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.|National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
mesh_terms: D020285:Cryoelectron Microscopy / Q000295:instrumentation*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation*; D008958:Models, Molecular; D023642:Rudiviridae / Q000737:chemistry / Q000648:ultrastructure*; D016944:Sulfolobus / Q000821:virology; D015678:Viral Structural Proteins / Q000737:chemistry / Q000648:ultrastructure; D014771:Virion / Q000737:chemistry / Q000648:ultrastructure
chemical_list: D015678:Viral Structural Proteins
keywords: Direct detectors; Electron microscopy; Near-atomic resolution; Sulfolobus turreted icosahedral virus
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