Departments of Biochemistry and Molecular Biophysics and of Biological Sciences, Columbia University, New York, New York, USA.
Nat Protoc. 2017 Feb;12(2):209-212. doi: 10.1038/nprot.2017.004. Epub 2017 Jan 5.
In single-particle cryo-electron microscopy (cryo-EM), molecules suspended in a thin aqueous layer are rapidly frozen and imaged at cryogenic temperature in the transmission electron microscope. From the random projection views, a three-dimensional image is reconstructed, enabling the structure of the molecule to be obtained. In this article I discuss technological progress over the past decade, which has, in my own field of study, culminated in the determination of ribosome structure at 2.5-Å resolution. I also discuss likely future improvements in methodology.
在单颗粒冷冻电子显微镜(cryo-EM)中,悬浮在薄水层中的分子被迅速冷冻,并在低温电子显微镜中以低温成像。从随机投影视图中,重建三维图像,从而获得分子的结构。在本文中,我讨论了过去十年中的技术进步,这些进步在我自己的研究领域中,最终导致了核糖体结构在 2.5-Å分辨率下的确定。我还讨论了方法学的未来可能改进。