通过倾斜处理单颗粒冷冻电镜中首选的样品取向问题。
Addressing preferred specimen orientation in single-particle cryo-EM through tilting.
作者信息
Tan Yong Zi, Baldwin Philip R, Davis Joseph H, Williamson James R, Potter Clinton S, Carragher Bridget, Lyumkis Dmitry
机构信息
The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, New York, USA.
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA.
出版信息
Nat Methods. 2017 Aug;14(8):793-796. doi: 10.1038/nmeth.4347. Epub 2017 Jul 3.
Abstract
We present a strategy for tackling preferred specimen orientation in single-particle cryogenic electron microscopy by employing tilts during data collection. We also describe a tool to quantify the resulting directional resolution using 3D Fourier shell correlation volumes. We applied these methods to determine the structures at near-atomic resolution of the influenza hemagglutinin trimer, which adopts a highly preferred specimen orientation, and of ribosomal biogenesis intermediates, which adopt moderately preferred orientations.
摘要
我们提出了一种在单颗粒低温电子显微镜中通过在数据收集过程中采用倾斜来解决首选标本取向问题的策略。我们还描述了一种使用三维傅里叶壳层相关体积来量化所得方向分辨率的工具。我们将这些方法应用于确定流感血凝素三聚体（其采用高度首选的标本取向）和核糖体生物合成中间体（其采用中等首选取向）的近原子分辨率结构。
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