Real-time phasor analysis of fast FLIM for visualizing metabolic dynamics

Fluorescence lifetime imaging microscopy (FLIM) of reduced nicotinamide adenine dinucleotide and reduced nicotinamide adenine dinucleotide phosphate, together termed NAD(P)H, is often used to analyze and characterize the metabolic state of biological samples. The use of photon counting in FLIM of NAD(P)H often leads to long accumulation times which average out sub-second dynamics. This challenge is overcome with the presented two-photon fast FLIM system that uses an analog detection system and fast digitizer, enabling imaging up to 20 frames per second. Furthermore, GPU processing is used for real-time phasor analysis and visualization of collected data.