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  In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate

Barton, B., Rhinow, D., Walter, A., Schröder, R., Benner, G., Majorovits, E., et al. (2011). In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate. Ultramicroscopy, 111, 1696-1705.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0024-D643-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0024-D644-8
Genre: Journal Article

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 Creators:
Barton, B.1, Author              
Rhinow, D.1, Author              
Walter, A.1, Author              
Schröder, R.2, Author
Benner, G., Author
Majorovits, E., Author
Matijevic, M., Author
Niebel, H., Author
Müller, H., Author
Haider, M., Author
Lacher, M., Author
Schmitz, S., Author
Holik, P., Author
Kühlbrandt, W.1, Author              
Affiliations:
1Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068291              
2Max Planck Society, ou_persistent13              

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Free keywords: Electron cryo-microscopy; Phase plate; Electrostatic Einzel lens; Aberration correction; Phase contrast; Macromolecular complexes
 Abstract: 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.

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Language(s): eng - English
 Dates: 2011
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 573931
 Degree: -

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Title: Ultramicroscopy
  Alternative Title : Ultramicroscopy
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 111 Sequence Number: - Start / End Page: 1696 - 1705 Identifier: -