Asynchronous-spectral fusion fluorescence microscopy for microsecond-scale behavioral dynamics

Abstract: Event-based image sensors provide microsecond temporal resolution but lack spectral discrimination, whereas diffractive spectral imagers encode wavelength information at conventional frame rates. We introduce a fluorescence microscopy architecture that fuses asynchronous event streams with diffraction-encoded CMOS measurements to decouple temporal and spectral sampling. The system achieves ~3.9 um spatial resolution over a 0.5 mm field of view, effective temporal resolution down to 100 us, and differentiates fluorophores whose emission peaks are separated by only 23 nm. By synchronizing and computationally merging both sensing modalities, we enable spectrally resolved tracking at 100,000 frames/s without scanning or filter switching.