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Towards high-throughput in situ structural biology using electron cryotomography
Progress in Biophysics and Molecular Biology ( IF 3.8 ) Pub Date : 2020-06-21 , DOI: 10.1016/j.pbiomolbio.2020.05.010
Jan Böhning 1 , Tanmay A M Bharat 1
Affiliation  

Electron cryotomography is a rapidly evolving method for imaging macromolecules directly within the native environment of cells and tissues. Combined with sub-tomogram averaging, it allows structural and cell biologists to obtain sub-nanometre resolution structures in situ. However, low throughput in cryo-ET sample preparation and data acquisition, as well as difficulties in target localisation and sub-tomogram averaging image processing, limit its widespread usability. In this review, we discuss new advances in the field that address these throughput and technical problems. We focus on recent efforts made to resolve issues in sample thinning, improvement in data collection speed at the microscope, strategies for localisation of macromolecules using correlated light and electron microscopy and advancements made to improve resolution in sub-tomogram averaging. These advances will considerably decrease the amount of time and effort required for cryo-ET and sub-tomogram averaging, ushering in a new era of structural biology where in situ macromolecular structure determination will be routine.



中文翻译:

使用电子冷冻断层扫描法实现高通量原位结构生物学

电子冷冻断层扫描是一种快速发展的方法,可直接在细胞和组织的天然环境中对大分子进行成像。结合亚断层扫描平均,它允许结构和细胞生物学家在原位获得亚纳米分辨率的结构. 然而,cryo-ET 样品制备和数据采集的低吞吐量,以及目标定位和亚断层图像平均图像处理的困难,限制了其广泛使用。在这篇综述中,我们讨论了该领域解决这些吞吐量和技术问题的新进展。我们专注于最近为解决样品稀释问题所做的努力、显微镜数据收集速度的提高、使用相关光和电子显微镜定位大分子的策略以及提高亚断层图像平均分辨率的进展。这些进步将大大减少冷冻 ET 和亚断层扫描平均所需的时间和精力,开启原位结构生物学的新时代 大分子结构测定将成为常规。

更新日期:2020-06-21
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