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J Struct Biol. 2018 Oct;204(1):80-84. doi: 10.1016/j.jsb.2018.07.007. Epub 2018 Jul 11.

A simple and robust procedure for preparing graphene-oxide cryo-EM grids.

Author information

1
Department of Biochemistry and Biophysics, University of California San Francisco, CA 94143, United States.
2
Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States.
3
Department of Biochemistry and Biophysics, University of California San Francisco, CA 94143, United States; Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States. Electronic address: agard@msg.ucsf.edu.
4
Department of Biochemistry and Biophysics, University of California San Francisco, CA 94143, United States; Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States. Electronic address: ycheng@ucsf.edu.

Abstract

Graphene oxide (GO) sheets have been used successfully as a supporting substrate film in several recent cryogenic electron-microscopy (cryo-EM) studies of challenging biological macromolecules. However, difficulties in preparing GO-covered holey carbon EM grids have limited their widespread use. Here, we report a simple and robust method for covering holey carbon EM grids with GO sheets and demonstrate that these grids can be used for high-resolution single particle cryo-EM. GO substrates adhere macromolecules, allowing cryo-EM grid preparation with lower specimen concentrations and provide partial protection from the air-water interface. Additionally, the signal of the GO lattice beneath the frozen-hydrated specimen can be discerned in many motion-corrected micrographs, providing a high-resolution fiducial for evaluating beam-induced motion correction.

KEYWORDS:

Cryo-EM; EM grids; Graphene-oxide; Motion correction

PMID:
30017701
PMCID:
PMC6119484
[Available on 2019-10-01]
DOI:
10.1016/j.jsb.2018.07.007

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