Metabolic imaging by NAD(P)H FLIM requires the decay functions in the individual pixels to be resolved into the decay components of bound and unbound NADH/NAD(P)H. Metabolic information is contained in the lifetime and relative amplitudes of the components. The separation of the decay components and the accuracy of the amplitudes and lifetimes improves substantially by using the ultra-fast HPM-100-06 and HPM-100-07 hybrid detectors.
The IRF width in combination with the SPC-150N and SPC-150NX TCSPC modules is less than 20 ps (FWHM) . An IRF this fast does not interfere with the fluorescence decay. The usual deconvolution process in the data analysis then virtually becomes a simple curve fitting, and the decay parameters are obtained at unprecedented accuracy.
Keywords: FLIM, NADH, Label-Free, Metabolic Imaging, Multi-Exponential Decay Analysis