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《中国化学会-生物物理化学专业委员会第四届全国生物物理化学会议论文集》2016年
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Scanning single-molecule fluorescence correlation spectroscopy enables kinetics study from microseconds to seconds

毕慧敏  
【摘要】:Single-molecule fluorescence measurements have been widely used to explore kinetics and dynamics of biological systems. Among them, single-molecule imaging(SMI) is good at tracking processes slower than tens of milliseconds, whereas fluorescence correlation spectroscopy(FCS) is good at probing processes faster than submilliseconds. However, there is still shortage of simple yet effective single-molecule fluorescence method to cover the time-scale between submilliseconds and tens of milliseconds. To effectively bridge this millisecond gap, we developed a single-molecule fluorescence correlation spectroscopy(smFCS) method that works on surface-immobilized single molecules through surface scanning. We validated it by monitoring the classical DNA hairpin folding process. With a wide time window from microseconds to seconds, the experimental data are well fitted to the two-state folding model. All relevant molecular parameters, including the relative fluorescence brightness, equilibrium constant, and reaction rate constants, were uniquely determined.
【作者单位】:北京大学化学与分子工程学院
【分类号】:O561

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