title: Automated batch fiducial-less tilt-series alignment in Appion using Protomo.
abstract: 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.
authors.affiliation: Department of Physics, 77 Chieftan Way, Florida State University, Tallahassee, FL 32306, USA.
authors.forename: Alex J
authors.initials: AJ
authors.lastname: Noble
authors.affiliation: Department of Chemistry and Biochemistry, 95 Chieftain Way, Florida State University, Tallahassee, FL 32306, USA; Institute of Molecular Biophysics, 91 Chieftan Way, Florida State University, Tallahassee, FL 32306, USA. Electronic address: sstagg@fsu.edu.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Stagg
mesh_terms: D057205:Automation, Laboratory / Q000379:methods; D020285:Cryoelectron Microscopy / Q000379:methods*; D000229:Dependovirus; D055032:Electron Microscope Tomography / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D012984:Software
keywords: Automated processing; Cryo-electron tomography; Marker-free tilt-series alignment; Protomo; Single particle tomography; Transmission electron microscopy