Functionalized graphene grids with various charges for single-particle cryo-EM

Ye Lu, Nan Liu (), Yongbo Liu, Liming Zheng, Junhao Yang, Jia Wang, Xia Jia, Qinru Zi, Hailin Peng (), Yu Rao () and Hong-Wei Wang ()
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Ye Lu: Tsinghua University
Nan Liu: Tsinghua University
Yongbo Liu: Tsinghua University
Liming Zheng: Peking University
Junhao Yang: Tsinghua University
Jia Wang: Tsinghua University
Xia Jia: Tsinghua University
Qinru Zi: Tsinghua University
Hailin Peng: Peking University
Yu Rao: Tsinghua University
Hong-Wei Wang: Tsinghua University

Nature Communications, 2022, vol. 13, issue 1, 1-8

Abstract: Abstract A major hurdle for single particle cryo-EM in structural determination lies in the specimen preparation impaired by the air-water interface (AWI) and preferential particle-orientation problems. In this work, we develop functionalized graphene grids with various charges via a dediazoniation reaction for cryo-EM specimen preparation. The graphene grids are paraffin-assistant fabricated, which appear with less contaminations compared with those produced by polymer transfer method. By applying onto three different types of macromolecules, we demonstrate that the high-yield charged graphene grids bring macromolecules away from the AWI and enable adjustable particle-orientation distribution for more robust single particle cryo-EM structural determination.

Date: 2022
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DOI: 10.1038/s41467-022-34579-w

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