High-yield monolayer graphene grids for near-atomic resolution cryoelectron microscopy

Yimo Han, Xiao Fan, Haozhe Wang, Fang Zhao, Christopher G. Tully, Jing Kong, Nan Yao, Nieng Yan

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Cryogenic electron microscopy (cryo-EM) has become one of the most powerful techniques to reveal the atomic structures and working mechanisms of biological macromolecules. New designs of the cryo-EM grids - aimed at preserving thin, uniform vitrified ice and improving protein adsorption - have been considered a promising approach to achieving higher resolution with the minimal amount of materials and data. Here, we describe a method for preparing graphene cryo-EM grids with up to 99% monolayer graphene coverage that allows for more than 70% grid squares for effective data acquisition with improved image quality and protein density. Using our graphene grids, we have achieved 2.6-Å resolution for streptavidin, with a molecular weight of 52 kDa, from 11, 000 particles. Our graphene grids increase the density of examined soluble, membrane, and lipoproteins by at least 5-fold, affording the opportunity for structural investigation of challenging proteins which cannot be produced in large quantity. In addition, our method employs only simple tools that most structural biology laboratories can access. Moreover, this approach supports customized grid designs targeting specific proteins, owing to its broad compatibility with a variety of nanomaterials.

Original languageEnglish (US)
Pages (from-to)1009-1014
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number2
DOIs
StatePublished - Jan 14 2020

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • Cryo-EM
  • Graphene grid
  • High resolution
  • Structure determination
  • UV/ozone

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