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Nat Methods. 2018 Oct;15(10):793-795. doi: 10.1038/s41592-018-0139-3. Epub 2018 Sep 24.

Reducing effects of particle adsorption to the air-water interface in cryo-EM.

Author information

1
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, USA.
2
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.
3
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, USA. bcarr@nysbc.org.
4
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA. bcarr@nysbc.org.

Abstract

Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt preferred orientations. By using a rapid plunge-freezing robot and nanowire grids, we were able to reduce some of the deleterious effects of the air-water interface by decreasing the dwell time of particles in thin liquid films. We demonstrated this by using single-particle cryo-EM and cryo-electron tomography (cryo-ET) to examine hemagglutinin, insulin receptor complex, and apoferritin.

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PMID:
30250056
PMCID:
PMC6168394
DOI:
10.1038/s41592-018-0139-3
[Indexed for MEDLINE]
Free PMC Article

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