biostudies-literatureUnknown7(3)Liu NAlthough single-particle cryogenic electron microscopy (cryo-EM) has been applied extensively for elucidating many crucial biological mechanisms at the molecular level, this technique still faces critical challenges, the major one of which is to prepare the high-quality cryo-EM specimen. Aiming to achieve a more reproducible and efficient cryo-EM specimen preparation, novel supporting films including graphene-based two-dimensional materials have been explored in recent years. Here we report a robust and simple method to fabricate EM grids coated with single- or few-layer reduced graphene oxide (RGO) membrane in large batch for high-resolution cryo-EM structural determination. The RGO membrane has decreased interlayer space and enhanced electrical conductivity in comparison to regular graphBiophysics reports227-238https://www.ebi.ac.uk/biostudies/studies/S-EPMC10244798biostudies-literatureReduced graphene oxide membrane as supporting film for high-resolution cryo-EM.PMC10244798Xu KZhang JLiu NWang JWang HWXu JPeng HHu CYang ZZheng LZhang XChen YGao XWang XCheng HLan JfalseReduced graphene oxide membrane as supporting film for high-resolution cryo-EM.Although single-particle cryogenic electron microscopy (cryo-EM) has been applied extensively for elucidating many crucial biological mechanisms at the molecular level, this technique still faces critical challenges, the major one of which is to prepare the high-quality cryo-EM specimen. Aiming to achieve a more reproducible and efficient cryo-EM specimen preparation, novel supporting films including graphene-based two-dimensional materials have been explored in recent years. Here we report a robust and simple method to fabricate EM grids coated with single- or few-layer reduced graphene oxide (RGO) membrane in large batch for high-resolution cryo-EM structural determination. The RGO membrane has decreased interlayer space and enhanced electrical conductivity in comparison to regular graph2021-01-01T00:00:00Z2021 Jun2025-04-21T13:39:43.044Z2025-04-21T13:39:43.044ZS-EPMC102447983728749010.52601/bpr.2021.210007