title: Movies of ice-embedded particles enhance resolution in electron cryo-microscopy.
abstract: Low-dose images obtained by electron cryo-microscopy (cryo-EM) are often affected by blurring caused by sample motion during electron beam exposure, degrading signal especially at high resolution. We show here that we can align frames of movies, recorded with a direct electron detector during beam exposure of rotavirus double-layered particles, thereby greatly reducing image blurring caused by beam-induced motion and sample stage instabilities. This procedure increases the efficiency of cryo-EM imaging and enhances the resolution obtained in three-dimensional reconstructions of the particle. Using movies in this way is generally applicable to all cryo-EM samples and should also improve the performance of midrange electron microscopes that may have limited mechanical stability and beam coherence.
authors.affiliation: National Resource for Automated Molecular Microscopy, Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Melody G
authors.initials: MG
authors.lastname: Campbell
authors.forename: Anchi
authors.initials: A
authors.lastname: Cheng
authors.forename: Axel F
authors.initials: AF
authors.lastname: Brilot
authors.forename: Arne
authors.initials: A
authors.lastname: Moeller
authors.forename: Dmitry
authors.initials: D
authors.lastname: Lyumkis
authors.forename: David
authors.initials: D
authors.lastname: Veesler
authors.forename: Junhua
authors.initials: J
authors.lastname: Pan
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Harrison
authors.forename: Clinton S
authors.initials: CS
authors.lastname: Potter
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Grigorieff
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D007053:Ice; D012401:Rotavirus / Q000648:ultrastructure*
chemical_list: D007053:Ice