Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy

Abstract

Rapid 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. Here we report on a computational light-sheet microscopy able to achieve minute-timescale high-resolution mapping of entire macro-scale organs. Through combining a dual-side confocally-scanned Bessel light-sheet illumination which provides thinner-and-wider optical sectioning of deep tissues, with a content-aware compressed sensing (CACS) computation pipeline which further improves the contrast and resolution based on a single acquisition, our approach yields 3D images with high, isotropic spatial resolution and rapid acquisition over two-order-of-magnitude faster than conventional 3D microscopy implementations. We demonstrate the imaging of whole brain (~400\u2009mm 3 ), entire gastrocnemius and tibialis muscles (~200\u2009mm 3 ) of mouse at ultra-high throughput of 5~10\u2009min per sample and post-improved subcellular resolution of ~\u20091.5\u2009μm (0.5-μm iso-voxel size). Various system-level cellular analyses, such as mapping cell populations at different brain sub-regions, tracing long-distance projection neurons over the entire brain, and calculating neuromuscular junction occupancy across whole muscle, are also readily accomplished by our method. High resolution imaging of large biological volumes typically takes a long time from hours to days. Here the authors use a Bessel light-sheet approach combined with a content-aware compressed sensing computational pipeline to image whole mouse organs at subcellular resolution in a few minutes.