Centre for Molecular Biology of Heidelberg University (ZMBH), 69120 Heidelberg, Germany.
Structural and Computational Biology Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
Curr Opin Struct Biol. 2018 Oct;52:111-118. doi: 10.1016/j.sbi.2018.08.009. Epub 2018 Oct 16.
Structural and cell biology have traditionally been separate disciplines and employed techniques that were well defined within the realm of either one or the other. Recent technological breakthroughs propelled electron microscopy of frozen hydrated specimens (cryo-EM) followed by single-particle analysis (SPA) to become a widely applied approach for obtaining near-atomic resolution structures of purified macromolecules. In parallel, ongoing developments on sample preparation are increasingly successful in bringing molecular views into cell biology. Cryo-electron tomography (cryo-ET) has so far served as the main imaging modality employed in these efforts towards obtaining three-dimensional (3D) volumes of heterogeneous molecular assemblies. We review the state-of-the-art in cryo-ET and computational processing and describe the current opportunities and frontiers for in-cell applications.
结构和细胞生物学传统上是两个独立的学科，所采用的技术在这两个领域内都有明确的定义。最近的技术突破推动了冷冻水合标本的电子显微镜（cryo-EM），随后是单颗粒分析（SPA），成为获得纯化大分子的近原子分辨率结构的广泛应用方法。与此同时，不断发展的样品制备技术在将分子视角引入细胞生物学方面越来越成功。到目前为止，冷冻电子断层扫描（cryo-ET）一直是这些努力中用于获取异质分子组装体三维（3D）体积的主要成像方式。我们回顾了 cryo-ET 和计算处理的最新技术，并描述了细胞内应用的当前机会和前沿。
