A new method for vitrifying samples for cryoEM.

Autor: Razinkov I; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Dandey V; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Wei H; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Zhang Z; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Melnekoff D; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Rice WJ; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Wigge C; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Potter CS; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center., Carragher B; Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Jazyk: angličtina
Zdroj: Journal of structural biology [J Struct Biol] 2016 Aug; Vol. 195 (2), pp. 190-198. Date of Electronic Publication: 2016 Jun 08.
DOI: 10.1016/j.jsb.2016.06.001
Abstrakt: Almost every aspect of cryo electron microscopy (cryoEM) has been automated over the last few decades. One of the challenges that remains to be addressed is the robust and reliable preparation of vitrified specimens of suitable ice thickness. We present results from a new device for preparing vitrified samples. The successful use of the device is coupled to a new "self-blotting" grid that we have developed to provide a method for spreading a sample to a thin film without the use of externally applied filter paper. This new approach has the advantage of using small amounts of protein material, resulting in large areas of ice of a well defined thickness containing evenly distributed single particles. We believe that these methods will in the future result in a system for vitrifying grids that is completely automated.
(Copyright © 2016 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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