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

By Eugene Palovcak, Feng Wang, Shawn Q. Zheng, Zanlin Yu, Sam Li, David Bulkley, David A. Agard, Yifan Cheng

Posted 27 Mar 2018
bioRxiv DOI: 10.1101/290197 (published DOI: 10.1016/j.jsb.2018.07.007)

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 its widespread use. Here, we report a simple and robust method for covering holey carbon EM grids with GO sheets and demonstrate that these grids are suitable for high-resolution single particle cryo-EM. GO substrates adhere macromolecules, allowing cryo-EM grid preparation with lower specimen concentrations and providing partial protection from the air-water interface. Additionally, the signal from images 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.

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