Improved sample dispersion in cryo-EM using "perpetually-hydrated" graphene oxide flakes

Martin Cheung; Hidehito Adaniya; Cathal Cassidy; Masao Yamashita; Kun-Lung Li; Seita Taba; Tsumoru Shintake

doi:10.1016/j.jsb.2018.03.006

Improved sample dispersion in cryo-EM using "perpetually-hydrated" graphene oxide flakes

J Struct Biol. 2018 Mar 15:S1047-8477(18)30083-2. doi: 10.1016/j.jsb.2018.03.006. Online ahead of print.

Authors

Martin Cheung¹, Hidehito Adaniya², Cathal Cassidy², Masao Yamashita², Kun-Lung Li², Seita Taba², Tsumoru Shintake²

Affiliations

¹ Quantum Wave Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, Kunigami District Okinawa Prefecture, 904-0412 Japan. Electronic address: martin.cheung@oist.jp.
² Quantum Wave Microscopy Unit, Okinawa Institute of Science and Technology Graduate University, Kunigami District Okinawa Prefecture, 904-0412 Japan.

PMID: 29551713
DOI: 10.1016/j.jsb.2018.03.006

Abstract

For many macromolecular complexes, the inability to uniformly disperse solubilized specimen particles within vitreous ice films precludes their analysis by cryo-electron microscopy (cryo-EM). Here, we introduce a sample preparation process using "perpetually-hydrated" graphene oxide flakes as particle support films, and report vastly improved specimen dispersion. Furthermore, we provide evidence that the presence of graphene oxide flakes in vitreous ice results in a significant reduction in electron beam-induced specimen decomposition. The new method introduced in this study incorporates hydrated graphene oxide flakes into a standard sample preparation regime, without the need for additional tools or devices, making it a cost-effective and easily adoptable alternative to currently available sample preparation approaches.

Keywords: Cryo-electron microscopy; Sample preparation; Vitreous ice; Vitrification.