High-throughput wide-field multispectral FLIM system based on a 16-channel silicon photomultiplier array

Fluorescence microscopy is nowadays one of the most diffused techniques to study photophysical processes and molecular interactions in both biomedical and material science fields. Since each fluorophore is characterized by a specific emission spectrum and lifetime, it is essential not only to spatially localize its position but also to measure its spectral and temporal properties with a multispectral fluorescence lifetime imaging microscope (lambda FLIM). Moreover, to study rapidly evolving samples, a measurement system capable of fast acquisitions is needed. lambda FLIM systems are currently limited on acquisition speed and cannot reach high throughput. In this work, we propose a novel wide-field multispectral fluorescence lifetime imaging microscope based on a 16-channel silicon photomultiplier (SiPM) array. Our system, thanks to the SiPM technology along with the single pixel camera and compress sensing approaches is able to acquire multidimensional measurements (space, spectrum, and time) at high frame rate. We validate the system on moving fluorescent samples capturing snapshots at a frame rate of up to 13 fps. The developed system could enable enhanced specificity in real-time fluorescence imaging in biological and biosensing applications.