Three-observation-window time-gated algorithm for fluorescence lifetime detection

Abstract

A novel three-observation-window time-gated algorithm that combines overlapped windows and discrete windows together is developed for accurate fluorescence lifetime extraction. The new algorithm adopting a rapid lifetime determination strategy can offer an excellent ability to precisely detect long fluorescence lifetime for fluorescence lifetime imaging microscopy. Monte Carlo simulation indicates that an extremely small relative standard deviation below 0.4% is obtained over a wide fluorescence lifetime range from 5 ns to 30 ns. The detection error of the short fluorescence lifetime less than 5 ns is further reduced by means of an adaptive window width method. In contrast to other algorithms, such as time-correlated single-photon counting and traditional gated-window methods, not only the detection range but also the measurement accuracy is dramatically enhanced.