Photophysics of fluorescent probes for single-molecule biophysics and super-resolution imaging.
Photophysics of fluorescent probes for single-molecule biophysics and super-resolution imaging.
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DOI：
10.1146/annurev-physchem-032210-103340
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发表时间：
2012
影响因子：
14.7
通讯作者：
Tinnefeld P
中科院分区：
文献类型：
--
作者：
Ha T;Tinnefeld P
单分子荧光光谱和超分辨率显微镜是正在进行的技术革命的重要组成部分，以前所未有的清晰度和精度揭示生物化学和细胞过程。对荧光团的光物理性质的要求是严格的，并驱动适当探针的选择。这种荧光团不是简单的特定颜色和亮度的灯泡，而是在光谱参数、氧化还原性质、大小和水溶性、光稳定性等方面具有自己的“个性”。在这里，我们回顾了荧光探针，有机荧光团和荧光蛋白，用于应用程序，如粒子跟踪，单分子FRET，化学计量测定，和超分辨率成像的荧光物理。特别令人感兴趣的是巯基诱导的Cy 5闪烁，这是单分子生物物理研究的诅咒，后来使用Trolox通过还原/氧化系统克服了这一诅咒，但由于可控的光开关，这对超分辨率成像是一个布恩。理解量子物理学对于设计和解释单分子实验至关重要。
Single-molecule fluorescence spectroscopy and super-resolution microscopy are important elements of the ongoing technical revolution to reveal biochemical and cellular processes in unprecedented clarity and precision. Demands placed on the photophysical properties of the fluorophores are stringent and drive the choice of appropriate probes. Such fluorophores are not simple light bulbs of certain color and brightness but instead have their own ‘personalities’ regarding spectroscopic parameters, redox properties, size and water solubility, photostability and several more. Here, we review the photophysics of fluorescent probes, both organic fluorophores and fluorescent proteins, used in applications such as particle tracking, single molecule FRET, stoichiometry determination, and super-resolution imaging. Of particular interest is the thiol-induced blinking of Cy5, a curse for single molecule biophysical studies which was later overcome using Trolox through reducing/oxidizing system, but a boon for super-resolution imaging due to the controllable photoswitching. Understanding photophysics is critical in design and interpreting single molecule experiments.
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影响因子：
48
作者：
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