The role of data processing for lifetime image scanning microscopy and beyond: an update

Confocal microscopy is an essential tool in many academic disciplines due to its intrinsic sectioning capability. It combines naturally with hardware to obtain additional contrasts such as information on the fluorescent lifetime. 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 – advanced 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 such a processing is well established for intensity imaging some challenges arise in the context of lifetime imaging - or more general - imaging that generates information in non-spatial data-dimensions. We discuss some of these challenges and give examples for a novel data processing approach that extends to many lifetime microscopy methods.