Computational 3D multispectral fluorescence lifetime microscopy

This work presents what we believe to be a novel 3D multispectral fluorescence lifetime microscope that combines structured illumination microscopy (SIM) and single-pixel camera (SPC) techniques within a computational framework. The approach allows simultaneous acquisition of steady-state high-resolution spatial data and and low-resolution spatial-temporal spectral data, which are fused using data fusion algorithms to generate a five-dimensional dataset. Compressive sensing has been exploited to accelerate SPC acquisition. Experimental validation with fluorescent beads and cellular samples demonstrates high fidelity in spatial, spectral, and lifetime data compared to ground truth. Ultimately, our system advances the fast and non-invasive measurements of volumetric fluorescent samples. It provides spectral and temporal information without increasing acquisition time and still offers reliable, high spatial resolution measurements.