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          Institute: MPI für Biophysik     Collection: Abt. Strukturbiologie     Display Documents



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ID: 573931.0, MPI für Biophysik / Abt. Strukturbiologie
In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate
Authors:Barton, B.; Rhinow, D.; Walter, A.; Schröder, R.; Benner, G.; Majorovits, E.; Matijevic, M.; Niebel, H.; Müller, H.; Haider, M.; Lacher, M.; Schmitz, S.; Holik, P.; Kühlbrandt, W.
Language:English
Date of Publication (YYYY-MM-DD):2011
Title of Journal:Ultramicroscopy
Journal Abbrev.:Ultramicroscopy
Volume:111
Start Page:1696
End Page:1705
Review Status:Peer-review
Audience:Experts Only
Abstract / Description:We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, Cs corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of <0.6nm/h at the specimen. Maximal in-focus phase contrast was obtained by applying voltages between 80 and 700 mV to the phase plate electrode. The phase plate allows for in-focus imaging of biological objects with a signal-to-noise of 5–10 at a resolution of 2–3 nm, as demonstrated for frozen- hydrated virus particles and purple membrane at liquid-nitrogen temperature.
Free Keywords:Electron cryo-microscopy; Phase plate;
Electrostatic Einzel lens; Aberration correction; Phase contrast; Macromolecular complexes
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für Biophysik/Abteilung Strukturbiologie
External Affiliations:Carl Zeiss NTS GmbH, D-73447 Oberkochen, Germany;
CEOS GmbH, Englerstr. 26, 69126 Heidleberg, Germany;
Caesar Research Center, Ludwig-Erhard-Allee 2, D-53175 Bonn, Germany
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