论文标题
通过荧光相关光谱和抗抗启示率测量光物理过渡速率
Measuring photophysical transition rates with fluorescence correlation spectroscopy and antibunching
论文作者
论文摘要
我们提出了一种新方法，该方法将微秒时间尺度上的荧光相关光谱（FCS）与纳秒时间尺度上的荧光抗启动测量结果结合在一起，以测量荧光分子的光物理速率常数。抗挑战测量值使我们能够量化FCS测量设置的共聚焦检测体积内荧光分子的平均激发率。对此值的了解使我们可以绝对的方式量化从FCS曲线的微秒时间衰减中的交叉率和三胞胎状态寿命。我们对该方法进行了理论分析，并估算了共聚焦检测量的所有数量（激发率，光物理速率）引起的最大偏差，并且我们表明，在大多数情况下，这种偏差小于5 \％。我们采用了测量广泛使用染料的光物理速率常数〜110和atto〜655。
We present a new method that combines fluorescence correlation spectroscopy (FCS) on the microsecond time scale with fluorescence antibunching measurements on the nanosecond time scale for measuring photophysical rate constants of fluorescent molecules. The antibunching measurements allow us to quantify the average excitation rate of fluorescent molecules within the confocal detection volume of the FCS measurement setup. Knowledge of this value allows us then to quantify, in an absolute manner, the intersystem crossing rate and triplet state lifetime from the microsecond temporal decay of the FCS curves. We present a theoretical analysis of the method and estimate the maximum bias caused by the averaging of all quantities (excitation rate, photophysical rates) over the confocal detection volume, and we show that this bias is smaller than 5\% in most cases. We apply the method for measuring the photophysical rate constants of the widely used dyes Rhodamine~110 and ATTO~655.
