title: Practical Experience with Hole-Free Phase Plates for Cryo Electron Microscopy.
abstract: Phase plate (PP) imaging has proven to be valuable in transmission cryo electron microscopy of unstained, native-state biological specimens. Many PP types have been described, however until the recent implementation of the "hole-free" phase plate (HFPP), imaging has been challenging. We found the HFPP to be simple to construct and to set up in the transmission electron microscopy, but care in implementing automated data collection is needed. Performance may be variable, both initially and over time, thus it is important to monitor and evaluate image quality by observing the power spectrum. We found that while some HFPPs gave transfer to high resolution without CTF oscillation, most reached high resolution when operated with modest defocus.
authors.affiliation: 1NY State Department of Health,Wadsworth Center,PO Box 509,Albany,NY 12201,USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Marko
authors.affiliation: 1NY State Department of Health,Wadsworth Center,PO Box 509,Albany,NY 12201,USA.
authors.forename: Chyongere
authors.initials: C
authors.lastname: Hsieh
authors.affiliation: 3Department of Materials Science and Engineering,Rensselaer Polytechnic Institute,Troy,NY 12180,USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Leith
authors.affiliation: 4Department of MCD Biology,University of Colorado Boulder,Boulder,CO 80309,USA.
authors.forename: David
authors.initials: D
authors.lastname: Mastronarde
authors.affiliation: 5JEOL USA,11 Dearborn Road,Peabody,MA 01960,USA.
authors.forename: Sohei
authors.initials: S
authors.lastname: Motoki
mesh_terms: D020285:Cryoelectron Microscopy / Q000295:instrumentation* / Q000592:standards*; D046529:Microscopy, Electron, Transmission
keywords: SerialEM; contrast-transfer function; cryo electron microscopy (cryo-EM); cryo electron tomography (cryo-ET); phase plate