High-throughput fluorescence microscopy using multi-frame motion deblurring

We demonstrate multi-frame motion deblurring for gigapixel wide-field fluorescence microscopy using fast slide scanning with coded illumination. Our method illuminates the sample with multiple pulses within each exposure, in order to introduce structured motion blur. By deconvolving this known motion sequence from the set of acquired measurements, we recover the object with up to 10× higher SNR than when illuminated with a single pulse (strobed illumination), while performing acquisition at 5× higher frame-rate than a comparable stop-and-stare method. Our coded illumination sequence is optimized to maximize the reconstruction SNR. We also derive a framework for determining when coded illumination is SNR-optimal in terms of system parameters such as source illuminance, noise, and motion stage specifications. This helps system designers to choose the ideal technique for high-throughput microscopy of very large samples.