Fluorescence lifetime image scanning microscopy

Simultaneous investigations of the spatial arrangement and relative composition of proteins are of crucial importance in live-cell imaging. It was recently shown that Image Scanning Microscopy (ISM) can double the confocal resolution. Additionally, fluorescence lifetime based imaging (FLIM) opens an additional dimension in multispecies identification and separation. We combine these two techniques using an array of single-photon detectors together with a multichannel TCSPC and time tagging system for time-resolved single photon detection via tens of channels. First results illustrate the spatial resolution improvement and the selection of suitable fluorescent dyes to allow for lifetime based multiplexing.