The role of data processing for lifetime image scanning microscopy

Confocal microscopy is an essential tool in many academic disciplines due to its intrinsic sectioning capability. It combines naturally with time-resolved single photon detectors and time-correlated single photon counting (TCSPC) devices. This has established it as the leading platform for time resolved investigation methods such as fluorescence lifetime imaging (FLIM). With the recent introduction of high performance SPAD-arrays confocal microscopes can be readily upgraded to image scanning microscopes (ISM), improving their performance noticeably. In this work we shed some light on another crucial ingredient behind these performance gains – the role of data processing. A suitable processing allows to leverage the improved spatial resolution to its full extent and improve the sectioning even for single 2d-data slices. While these techniques are well established for intensity imaging some challenges arise in the context of lifetime imaging. We discuss these challenges and give a first outlook for a novel data processing approach that extends to many lifetime microscopy methods.