Development of a high-speed line-scanning fluorescence lifetime imaging microscope for biological imaging.

We report the development of a novel line-scanning microscope capable of acquiring high-speed time-correlated single-photon counting (TCSPC)-based fluorescence lifetime imaging microscopy (FLIM) imaging. The system consists of a laser-line focus, which is optically conjugated to a 1024 × 8 single-photon avalanche diode (SPAD)-based line-imaging complementary metal-oxide semiconductor (CMOS), with 23.78 µm pixel pitch at 49.31% fill factor. Incorporation of on-chip histogramming on the line-sensor enables acquisition rates 33 times faster than our previously reported bespoke high-speed FLIM platforms. We demonstrate the imaging capability of the high-speed FLIM platform in a number of biological applications.