Palovcak, E.; Wang, F.; Zheng, S. Q.; Yu, Z.; Li, S.; Betegon, M.; Bulkley, D.; Agard, D. A. & Cheng, Y. A simple and robust procedure for preparing graphene-oxide cryo-EM grids. J. Structural Biology, 2018, 204, 80-84
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.