title: Using the Volta phase plate with defocus for cryo-EM single particle analysis.
abstract: Previously, we reported an in-focus data acquisition method for cryo-EM single-particle analysis with the Volta phase plate (Danev and Baumeister, 2016). Here, we extend the technique to include a small amount of defocus which enables contrast transfer function measurement and correction. This hybrid approach simplifies the experiment and increases the data acquisition speed. It also removes the resolution limit inherent to the in-focus method thus allowing 3D reconstructions with resolutions better than 3 Å.
authors.affiliation: Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Radostin
authors.identifier: 0000-0001-6406-8993
authors.initials: R
authors.lastname: Danev
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Dimitry
authors.identifier: 0000-0001-7019-3221
authors.initials: D
authors.lastname: Tegunov
authors.affiliation: Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Baumeister
mesh_terms: D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods*
keywords: biophysics; cryo-EM; none; phase plate; proteasome; structural biology