High-speed isotropic-resolution axially swept light-sheet microscopy

Light-sheet fluorescence microscopy (LSFM) is a rapidly evolving imaging technology which is revolutionizing biomedical imaging due to its faster imaging speed and gentler illumination scheme. Lately, a variant of LSFM, called Axially swept light-sheet microscope (ASLM) has emerged as a promising technique: not only does it offer isotropic (XYZ) sub-micron resolution in tissues, it can also be scaled to acquire large field-of-view. However, ASLM suffers from two fundamental problems: lower detection signal and low achievable frame-rate which limit the choice of ASLM in several cases. Recently, we showed that simultaneous scanning two stacked light-sheets not only increased the imaging speed but also doubled ASLM's signal performance. Here we push this idea further and propose a simple construction of dual-focus ASLM that offers better control of the focus position which ultimately pushes the imaging speed of ASLM from 10 frames-per-second (FPS) to 40 FPS.