4.4 Article

Automated batch fiducial-less tilt-series alignment in Appion using Protomo

Journal

JOURNAL OF STRUCTURAL BIOLOGY
Volume 192, Issue 2, Pages 270-278

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2015.10.003

Keywords

Cryo-electron tomography; Marker-free tilt-series alignment; Single particle tomography; Automated processing; Transmission electron microscopy; Protomo

Funding

  1. National Institutes of Health [GM086892]
  2. Department of Physics at Florida State University

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The field of electron tomography has benefited greatly from manual and semi-automated approaches to marker-based tilt-series alignment that have allowed for the structural determination of multitudes of in situ cellular structures as well as macromolecular structures of individual protein complexes. The emergence of complementary metal-oxide semiconductor detectors capable of detecting individual electrons has enabled the collection of low dose, high contrast images, opening the door for reliable correlation-based tilt-series alignment. Here we present a set of automated, correlation-based tilt-series alignment, contrast transfer function (CTF) correction, and reconstruction workflows for use in conjunction with the Appion/Leginon package that are primarily targeted at automating structure determination with cryogenic electron microscopy. (C) 2015 Elsevier Inc. All rights reserved.

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