Uniform thin ice on ultraflat graphene for high-resolution cryo-EM

Liming Zheng, Nan Liu, Xiaoyin Gao, Wenqing Zhu, Kun Liu, Cang Wu, Rui Yan, Jincan Zhang, Xin Gao, Yating Yao, Bing Deng, Jie Xu, Ye Lu, Zhongmin Liu, Mengsen Li, Xiaoding Wei, Hong Wei Wang, Hailin Peng

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has become one of the main challenges for high-resolution cryo-EM. Here we found that the uniformity of thin ice relies on the surface flatness of the supporting film, and developed a method to use ultraflat graphene (UFG) as the support for cryo-EM specimen preparation to achieve better control of vitreous ice thickness. We show that the uniform thin ice on UFG improves the image quality of vitrified specimens. Using such a method we successfully determined the three-dimensional structures of hemoglobin (64 kDa), α-fetoprotein (67 kDa) with no symmetry, and streptavidin (52 kDa) at a resolution of 3.5 Å, 2.6 Å and 2.2 Å, respectively. Furthermore, our results demonstrate the potential of UFG for the fields of cryo-electron tomography and structure-based drug discovery.
Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalNature Methods
Volume20
Issue number1
DOIs
StatePublished - Jan 1 2023
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Biotechnology

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