KEYWORDS: Fluorescence lifetime imaging, Sensors, Luminescence, Photodetectors, Signal detection, Monte Carlo methods, Single photon, Analog electronics, Physics, Temporal resolution
GaAsP hybrid detectors, which is new kind of photodetector, has been known as its excellent performance in time correlated single photon counting technique. We have verified that this detector also shows excellent performance in analog mean-delay method, which is another kind of time-domain FLIM, so one can expect enhancement of performance in time-domain FLIM when using the hybrid detector.
Phasor plot analysis is one of the most powerful analysis technique in fluorescence lifetime imaging microscopy, especially for analysis of heterogeneous mixtures. Compared to frequency domain fluorescence lifetime measurement, time domain measurement offers information in various frequencies at once measurement, but needs high frequency sampling for stable signal acquisition, which requires a lot of memory in hardware and a long time for analysis, furthermore in TCSPC, acquisition time is extremely long due to low photon count rate. We suggest a new system with low pass filter, which leads to about 100 times faster measurement speed while maintaining precision and accuracy in usual modulation frequency.
KEYWORDS: Luminescence, Fluorescence lifetime imaging, Analog electronics, Signal processing, Signal detection, Super resolution, Photodetectors, Convolution, Microscopy, Fluorescence resonance energy transfer
Fluorescence lifetime imaging microscopy (FLIM) is a powerful imaging tool widely used in monitoring cells, organelles, and tissues in biosciences. Since fluorescence lifetimes of most probes are a few nanoseconds, 20 ps measurement resolution is normally required. This requirement is quite challenging even with the fastest available optical and electronic devices, and several brilliant time-domain super-resolution techniques have been proposed for FLIM. The analog mean-delay (AMD) method is a recently introduced time-domain super-resolution technique for FLIM. Detailed constraints in the AMD method and their impact on the performance of the AMD super-resolution lifetime measurement are presented with experiments and simulations.
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