Department of Biotechnology & Biophysics, Julius-Maximilian-University of Würzburg , 97074 Würzburg, Germany.
Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt , 60438 Frankfurt, Germany.
Chem Rev. 2017 Jun 14;117(11):7478-7509. doi: 10.1021/acs.chemrev.6b00667. Epub 2017 Mar 13.
Super-resolution fluorescence imaging by photoactivation or photoswitching of single fluorophores and position determination (single-molecule localization microscopy, SMLM) provides microscopic images with subdiffraction spatial resolution. This technology has enabled new insights into how proteins are organized in a cellular context, with a spatial resolution approaching virtually the molecular level. A unique strength of SMLM is that it delivers molecule-resolved information, along with super-resolved images of cellular structures. This allows quantitative access to cellular structures, for example, how proteins are distributed and organized and how they interact with other biomolecules. Ultimately, it is even possible to determine protein numbers in cells and the number of subunits in a protein complex. SMLM thus has the potential to pave the way toward a better understanding of how cells function at the molecular level. In this review, we describe how SMLM has contributed new knowledge in eukaryotic biology, and we specifically focus on quantitative biological data extracted from SMLM images.
通过单荧光团的光激活或光转换实现的超分辨率荧光成像和位置确定（单分子定位显微镜，SMLM）提供了具有亚衍射空间分辨率的显微镜图像。这项技术使人们能够更深入地了解蛋白质在细胞环境中的组织方式，其空间分辨率接近实际的分子水平。SMLM 的一个独特优势是，它提供了分子分辨率的信息，以及细胞结构的超分辨率图像。这允许对细胞结构进行定量访问，例如，了解蛋白质的分布和组织方式，以及它们与其他生物分子的相互作用方式。最终，甚至有可能确定细胞中的蛋白质数量和蛋白质复合物中的亚基数量。因此，SMLM 有可能为更好地理解细胞在分子水平上的功能铺平道路。在这篇综述中，我们描述了 SMLM 如何为真核生物学提供新知识，并且我们特别关注从 SMLM 图像中提取的定量生物学数据。
