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Functionalized graphene grids with various charges for single-particle cryo-EM

A major hurdle for single particle cryo-EM in structural determination lies in the specimen preparation impaired by the air-water interface (AWI) and preferential particle-orientation problems. In this work, we develop functionalized graphene grids with various charges via a dediazoniation reaction...

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Detalles Bibliográficos
Autores principales: Lu, Ye, Liu, Nan, Liu, Yongbo, Zheng, Liming, Yang, Junhao, Wang, Jia, Jia, Xia, Zi, Qinru, Peng, Hailin, Rao, Yu, Wang, Hong-Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9640669/
https://www.ncbi.nlm.nih.gov/pubmed/36344519
http://dx.doi.org/10.1038/s41467-022-34579-w
Descripción
Sumario:A major hurdle for single particle cryo-EM in structural determination lies in the specimen preparation impaired by the air-water interface (AWI) and preferential particle-orientation problems. In this work, we develop functionalized graphene grids with various charges via a dediazoniation reaction for cryo-EM specimen preparation. The graphene grids are paraffin-assistant fabricated, which appear with less contaminations compared with those produced by polymer transfer method. By applying onto three different types of macromolecules, we demonstrate that the high-yield charged graphene grids bring macromolecules away from the AWI and enable adjustable particle-orientation distribution for more robust single particle cryo-EM structural determination.