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Uniform thin ice on ultraflat graphene for high-resolution cryo-EM

Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has...

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Detalles Bibliográficos
Autores principales: Zheng, Liming, Liu, Nan, Gao, Xiaoyin, Zhu, Wenqing, Liu, Kun, Wu, Cang, Yan, Rui, Zhang, Jincan, Gao, Xin, Yao, Yating, Deng, Bing, Xu, Jie, Lu, Ye, Liu, Zhongmin, Li, Mengsen, Wei, Xiaoding, Wang, Hong-Wei, Peng, Hailin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group US 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834055/
https://www.ncbi.nlm.nih.gov/pubmed/36522503
http://dx.doi.org/10.1038/s41592-022-01693-y
Descripción
Sumario:Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has become one of the main challenges for high-resolution cryo-EM. Here we found that the uniformity of thin ice relies on the surface flatness of the supporting film, and developed a method to use ultraflat graphene (UFG) as the support for cryo-EM specimen preparation to achieve better control of vitreous ice thickness. We show that the uniform thin ice on UFG improves the image quality of vitrified specimens. Using such a method we successfully determined the three-dimensional structures of hemoglobin (64 kDa), α-fetoprotein (67 kDa) with no symmetry, and streptavidin (52 kDa) at a resolution of 3.5 Å, 2.6 Å and 2.2 Å, respectively. Furthermore, our results demonstrate the potential of UFG for the fields of cryo-electron tomography and structure-based drug discovery.