Khavnekar S, Wan W, Majumder P, Wietrzynski W, Erdmann P S, Plitzko J M
MPI for Biochemistry, Germany.
Vanderbilt University, United States.
J Struct Biol. 2023 Mar;215(1):107911. doi: 10.1016/j.jsb.2022.107911. Epub 2022 Nov 4.
Cryo-electron tomography (cryo-ET) and subtomogram averaging (STA) can resolve protein complexes at near atomic resolution, and when combined with focused ion beam (FIB) milling, macromolecules can be observed within their native context. Unlike single particle acquisition (SPA), cryo-ET can be slow, which may reduce overall project throughput. We here propose a fast, multi-position tomographic acquisition scheme based on beam-tilt corrected beam-shift imaging along the tilt axis, which yields sub-nanometer in situ STA averages.
冷冻电子断层扫描(cryo-ET)和亚断层平均(STA)能够以接近原子分辨率解析蛋白质复合物,并且与聚焦离子束(FIB)铣削相结合时,可以在其天然环境中观察大分子。与单颗粒采集(SPA)不同,cryo-ET可能速度较慢,这可能会降低整体项目通量。我们在此提出一种基于沿倾斜轴进行束倾斜校正的束移位成像的快速多位置断层扫描采集方案,该方案可产生亚纳米原位STA平均值。