Theveny Liam M, Mageswaran Shrawan Kumar, Chen William David, Martinez Matthew, Guérin Amandine, Chang Yi-Wei
Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Trends Parasitol. 2022 May;38(5):365-378. doi: 10.1016/j.pt.2022.01.006. Epub 2022 Feb 8.
Cryo-electron tomography (cryo-ET) is a cryo-electron microscopy (EM) approach that allows 3D imaging of cellular structures in near-native, frozen-hydrated conditions with molecular resolution. Continued development of technologies, including direct electron detectors, phase plates, and energy filters, has improved the information yield from cellular samples, which is further extended by newly developed workflows for data collection and analyses. Moreover, advanced sample-thinning techniques, such as cryogenic focused ion-beam (cryo-FIB) milling, provide access to parasitic events and structures that were previously inaccessible for cryo-ET. Cryo-ET has therefore become more versatile and capable of transforming our understanding of parasite biology, particularly that of apicomplexans. This review discusses cryo-ET's implementation, its recent contributions, and how it can reveal pathogenesis mechanisms in the near future using apicomplexans as a case study.
冷冻电子断层扫描(cryo-ET)是一种冷冻电子显微镜(EM)方法,它能够在接近天然的冷冻水合条件下以分子分辨率对细胞结构进行三维成像。包括直接电子探测器、相位板和能量过滤器在内的技术不断发展,提高了细胞样本的信息产出,新开发的数据采集和分析工作流程进一步扩展了这些信息。此外,先进的样品减薄技术,如低温聚焦离子束(cryo-FIB)铣削,能够观察到以前冷冻电子断层扫描无法观察到的寄生事件和结构。因此,冷冻电子断层扫描变得更加通用,能够改变我们对寄生虫生物学的理解,特别是对顶复门原虫的理解。本综述讨论了冷冻电子断层扫描的应用、其近期贡献,以及以顶复门原虫为例,它如何在不久的将来揭示发病机制。