Realization of a time-correlated photon counting technique for fluorescence analysis.

An ultralow level light detection module, the time-correlated photon counter, is proposed and evaluated for fluorescence analysis. The time-correlated photon counter employs a silicon photomultiplier as a photon counting sensor in conjunction with a Poisson statistics algorithm and a double time windows technique, and therefore it can accurately count the photon number. The time-correlated photon counter is compatible with the time-correlated single photon counting technique and can record the arrival time of very faint light signals. This low-cost and compact instrument was used to analyze the intensity and lifetime of fluorescein isothiocyanate; a limit of detection of 16 pg/ml with a large linear dynamic range from 2.86 pg/ml to 0.5 µg/ml was obtained, and the lifetime of fluorescein isothiocyanate was measured to be 3.758 ns, which agrees well with the results of a sophisticated commercial fluorescence analysis instrument. The time-correlated photon counter may be useful in applications such as point-of-care testing.