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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