Sensitivity analysis-based time gate selection procedure for biexponential fluorescence imaging

Fluorescence lifetime imaging is an essential analytical tool in studies of cells and tissues. Recently, translation of the technique from microscopy to in vivo or with augmented data (hyperspectral) is being developed. One major concern for these new implementations reside in the lengthy acquisition times required. Data reduction techniques are one approach to significantly reduce the imaging protocol times. Herein, a sensitivity analysis framework is developed for the multifaceted, high-dimensional problems of current and future applications of fluorescence imaging. This framework is applied to the temporal dimension of lifetime-based Förster resonance energy transfer imaging. Synthetic data is used to show acquisition time can be reduced by an order of magnitude with almost no decrease in parameter estimation accuracy.