Photostable and photoswitching fluorescent dyes for super-resolution imaging
Photostable and photoswitching fluorescent dyes for super-resolution imaging
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DOI：
10.1007/s00775-016-1435-y
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发表时间：
2017-07-01
影响因子：
3
通讯作者：
Kikuchi, Kazuya
中科院分区：
文献类型：
--
作者：
Minoshima, Masafumi;Kikuchi, Kazuya
超分辨荧光显微镜是近年来发展起来的一种用于生物研究的成像工具。已经开发了几种方法用于检测亚衍射限制区域中分子的荧光信号，打破了传统光学显微镜的衍射限制，并允许可视化细胞中详细的大分子结构。由于物镜在光学系统中暴露于强激光，即使在恶劣的光照射条件下，用于超分辨率显微镜的荧光团也必须被耐受。对于基于单分子定位的超分辨率显微镜，荧光团还必须是可光活化和可光开关的，因为活性荧光团的数量必须通过光照射来控制。这导致了越来越多的兴趣，在这些性质的荧光团的发展。在这篇简短的综述中，我们重点介绍了用于超分辨率显微镜的光稳定和光开关荧光染料的发展。本文介绍了近年来荧光团光稳定性的改善和基于光化学和光物理过程的荧光团光开关行为的控制。理解和操纵激发荧光团中的化学反应可以开发出适用于超分辨率成像应用的高度光稳定和高效光可切换的荧光团。
Super-resolution fluorescence microscopy is a recently developed imaging tool for biological researches. Several methods have been developed for detection of fluorescence signals from molecules in a subdiffraction-limited area, breaking the diffraction limit of the conventional optical microscopies and allowing visualization of detailed macromolecular structures in cells. As objectives are exposed to intense laser in the optical systems, fluorophores for super-resolution microscopy must be tolerated even under severe light irradiation conditions. The fluorophores must also be photoactivatable and photoswitchable for single-molecule localization-based super-resolution microscopy, because the number of active fluorophores must be controlled by light irradiation. This has led to growing interest in these properties in the development of fluorophores. In this mini-review, we focus on the development of photostable and photoswitching fluorescent dyes for super-resolution microscopy. We introduce recent efforts, including improvement of fluorophore photostability and control of photoswitching behaviors of fluorophores based on photochemical and photophysical processes. Understanding and manipulation of chemical reactions in excited fluorophores can develop highly photostable and efficiently photoswitchable fluorophores that are suitable for super-resolution imaging applications.