Fast volumetric fluorescence lifetime imaging of multicellular systems using single-objective light-sheet microscopy
Year: 2025
Type: article
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 demonstrate with lifetime-based multiplexing in 3D and time-lapse FLIM of tension probes on living embryonic organoids. We benchmark both scanned and static light-sheet modalities to facilitate adding FLIM capability to a large variety of light-sheet microscopes.
Source: Communications Biology
Institutions: Centre National de la Recherche Scientifique, Aix-Marseille Université, Laboratoire d’Informatique et Systèmes, Institut de Biologie du Développement Marseille, Charité - Universitätsmedizin Berlin, University of Nottingham, PicoQuant (Germany), University of Birmingham, Université de Bordeaux, Institut Interdisciplinaire de Neuroscience, National University of Singapore, Mechanobiology Institute, Bridging Mechanobiology and Medicine, École Polytechnique Fédérale de Lausanne
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,190
Awards: A high content screening platform for live imaging and deep learning-assisted quantitative assessment of 3D cell culture models of hepatotoxicity, G2450198060, G3010527862, G381665449, G4219922975, G5549015088, G6069939831, G7240651115, G7867670471, Integration of biochemical and mechanical signals at cell adhesive contacts during mammalian morphogenesis: a quantitative approach using a minimal self-organized system, G996255883
OpenAlex ID: W4416809061