Multi-focus light-field microscopy for high-speed large-volume imaging

Abstract High-speed visualization of three-dimensional (3D) processes across a large field view with cellular resolution is essential for understanding living systems. Light-field microscopy (LFM) has emerged as powerful tool fast volumetric imaging. However, one inherent limitation LFM that the achievable lateral degrades rapidly increase distance from focal plane, which hinders applications in observing thick samples. Here, we propose Spherical-Aberration-assisted scanning (SAsLFM), hardware-modification-free method modulates phase-space point-spread-functions (PSFs) to extend effective high-resolution range along z-axis by ~ 3 times. By transferring foci different depths, take full advantage redundant light-field data preserve finer details over an extended depth and reduce artifacts near original plane. Experiments on USAF-resolution chart zebrafish vasculatures were conducted verify effectiveness method. We further investigated capability SAsLFM dynamic samples imaging large-scale calcium transients mouse brain, tracking freely-moving jellyfish, recording development Drosophila embryos. In addition, combined deep-learning approaches, accelerated reconstruction three orders magnitude. Our compatible various techniques without increasing system complexity can facilitate high-speed