Using fluorescence lifetime for discriminating detector afterpulsing in fluorescence-correlation spectroscopy

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.