Advanced calibration and processing of a time-resolved fluorescence spectroscopy system

While conventional fluorescence spectroscopy is concerned with the spectral features of fluorescence emission, time-resolved fluorescence spectroscopy (TRFS) also captures the temporal behaviour allowing obtaining spectro-temporal fluorescence waveforms of the excited molecules, unique to each (organic) molecule. Fluorescence lifetime depends on both radiative (i.e. fluorescence) and non-radiative (i.e. quenching, FRET) processes. Therefore, TRFS may also provide information about pH, ion concentrations or oxygen in the local environment of fluorophores, important in tissue assessment. This work introduces a full calibration procedure of a compact system that integrates a miniaturized spectrograph layout with a high-speed time-gated image sensor employing Current-Assisted Photonic Sampler (CAPS) technology. In addition, enhancing the spectral and temporal characteristics of the measured fluorescence, spectrum and decay, using deconvolution techniques to correct for the point spread function (PSF) and the instrument response function (IRF) of the system.