Atomically Thin Bilayer Janus Membranes for Cryo-electron Microscopy

LM Zheng and N Liu and Y Liu and N Li and JC Zhang and CZ Wang and WQ Zhu and YA Chen and DC Ying and J Xu and Z Yang and XY Gao and JL Tang and XG Wang and ZB Liang and RQ Zou and YZ Li and P Gao and XD Wei and HW Wang and HL Peng, ACS NANO, 15, 16562-16571 (2021).

DOI: 10.1021/acsnano.1c06233

Cryo-electron microscopy (cryo-EM) has emerged as a vital tool to reveal the native structure of beam-sensitive biomolecules and materials. Yet high-resolution cryo-EM analysis is still limited by the poorly controlled specimen preparation and urgently demands a robust supporting film material to prepare desirable samples. Here, we developed a bilayer Janus graphene membrane with the top-layer graphene being functionalized to interact with target molecules on the surface, while the bottom layer being kept intact to reinforce its mechanical steadiness. The ultraclean and atomically thin bilayer Janus membrane prepared by our protocol on one hand generates almost no extra noise and on the other hand reduces the specimen motion during cryo-EM imaging, thus allowing the atomic- resolution characterization of surface functional groups. Using such Janus membranes in cryo-EM specimen preparation, we were able to directly image the lithium dendrite and reconstruct macromolecules at near-atomic resolution. Our results demonstrate the bilayer Janus design as a promising supporting material for high-resolution cryo-EM and EM imaging.

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