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FLIM principles. Schematic overview of fluorescence lifetime data acquisition and analysis in time−domain (td) and frequency−domain (fd) modes. In tdFLIM, photon arrival times are recorded after each pulsed excitation to build a decay histogram, which can be analyzed by multi-exponential curve fitting or transformed into the phasor space for model-free lifetime extraction. In fdFLIM, the emission is measured under high-frequency intensity modulation, yielding phase shift and demodulation factors that can likewise be processed by fitting or phasor analysis. Both approaches ultimately render lifetime maps as pseudocolor images for biological interpretation.
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HIGHER EDUCATON PRESS
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