Camera-based time-correlated single photon counting (TCSPC) is a method where the position and the arrival time of the photons are recorded simultaneously. This has some advantages for fluorescence lifetime imaging (FLIM) with certain types of microscopy. It also allows FLIM with a macroscopic field of view. We report on nanosecond fluorescence lifetime imaging (FLIM) combined with total internal reflection fluorescence (TIRF) microscopy based on a 40 mm diameter crossed delay line anode detector. It has a few 100 picoseconds time resolution, and is read out via three standard TCSPC boards. We apply this wide-field TCSPC detector to identify Förster Resonance Energy Transfer (FRET) in cell membrane proteins in TIR-FLIM microscopy. In addition, we use a TCSPC single photon avalanche diode (SPAD) array as a macroscopic camera to visualise varnish removal on paintings.
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Suhling et al. (2024) studied this question.
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