Fast volumetric fluorescence lifetime imaging of multicellular systems using single-objective light-sheet microscopy.

ABSTRACT: Fluorescence lifetime imaging (FLIM) is widely used for functional and multiplexed bioimaging. The lifetime of autofluorescence or fluorescent sensors encodes physiologically relevant parameters. Thus, FLIM is especially relevant for the investigation of living systems. However, application of FLIM to live specimen is hampered by its slow speed and high phototoxicity. To enable faster and gentler FLIM, we integrated single-objective light-sheet microscopy with pulsed excitation and time-resolved detection on a novel SPAD array detector. We achieved 10-100-fold acceleration compared to confocal FLIM, down to 100 ms acquisition time per image, with excellent quantitative agreement. The massively enhanced speed enables volumetric FLIM acquisitions on live multicellular specimens, which we dem

SUBMITTER: Dunsing-Eichenauer V 

PROVIDER: S-EPMC12715196 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature