Using fluorescence lifetime for discriminating detector afterpulsing in fluorescence-correlation spectroscopy
Using fluorescence lifetime for discriminating detector afterpulsing in fluorescence-correlation spectroscopy
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DOI：
10.1063/1.1863399
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发表时间：
2005-03-01
影响因子：
1.6
通讯作者：
Gregor, I
中科院分区：
文献类型：
--
作者：
Enderlein, J;Gregor, I
荧光相关光谱技术在物理、化学和生物学等领域有着广泛的应用。通常，FCS用工作在光子计数盖革模式下的灵敏光探测器测量。这种探测器的一个共同特性是后脉冲：在真正的光子探测之后产生虚假的光子探测事件。这样的后脉冲导致显着偏离测量的自相关函数从它的真值在短时间尺度上，并可以严重影响导出的参数快速过程，如三重态电子物理。在这里，我们详细讨论了后脉冲对FCS的影响。本文提出了一种新的消除后脉冲效应的方法，即利用时间相关单光子计数法从后脉冲事件中分离出真实的荧光信号。(C)2005年美国物理学会。
Fluorescence correlation spectroscopy (FCS) has become an important and widely used technique for many applications in physics, chemistry, and biology. Usually, FCS is measured with sensitive light detectors working in the photon-counting Geiger mode. A common property of such detectors is afterpulsing: the generation of spurious photon detection events after a genuine photon detection. Such afterpulsing causes a significant deviation of the measured autocorrelation function from its true value on a short time scale and can seriously influence derived parameters for fast processes such as triplet-state photophysics. Here, we discuss the impact of afterpulsing on FCS in detail. A new method is developed to eliminate afterpulsing effects by using time-correlated single-photon counting for separating the true fluorescence signal from afterpulsing events. (C) 2005 American Institute of Physics.