Using a self-developed single objective light-sheet microscopy set-up, Valentin Dunsing-Eichenauer from Aix-Marseille Université & CNRS, Johan Hummert from PicoQuant, Ivan Michel Antolović from Pi Imaging Technology, and colleagues show lifetime-based multiplexing in 3D as well as time-lapse FLIM of mechanosensitive tension probes at record speed in living embryonic organoids. The set-up is based on pulsed excitation using a high power laser prototype and time-resolved detection on a newly available SPAD array detector.
Confocal FLIM is a popular tool for multiplexing and functional imaging of sensor fluorophores, but it is limited with regard to acquisition speed of large samples and live cell compatibility. Light-sheet microscopy, on the other hand, allows for fast and gentle imaging of large specimen. Combining both techniques opens new possibilities for fast volumetric FLIM.