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


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.

SUBMITTER: Palovcak E 

PROVIDER: S-EPMC6119484 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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

Palovcak Eugene E   Wang Feng F   Zheng Shawn Q SQ   Yu Zanlin Z   Li Sam S   Betegon Miguel M   Bulkley David D   Agard David A DA   Cheng Yifan Y  

Journal of structural biology 20180711 1


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  ...[more]

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