Fluorescence lifetime imaging through scattering media

Fluorescence lifetime determination has proven to be useful, e.g. identification of molecules, quantitative estimation of species concentration and determination of temperatures. Lifetime determination of exponentially decaying signals is challenging if signals of different decay rates are being mixed, resulting in erroneous results. Such issues occur when the contrast of the measurement object is low, which can be limiting in applied measurements due to spurious light scattering. A solution is presented here where structured illumination is used to enhance image contrast in fluorescence lifetime wide-field imaging. Lifetime imaging determination was carried out using Dual Imaging Modeling Evaluation (DIME), and spatial lock-in analysis was used for removing spurious scattered signal to enable fluorescence lifetime imaging through scattering media.