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 mm3), entire gastrocnemius and tibialis muscles (~200 mm3) of mouse at ultra-high throughput of 5~10 min per sample and post-improved subcellular resolution of ~ 1.5 μm (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.
Highlights
Rapid 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science
To extract the various cellular profiles of different physiological functions within specimens, three-dimensional (3D) highresolution imaging is required throughout a mesoscale-sized volume. Creating such a large-scale image dataset has posed significant challenges for conventional 3D light microscopy methods, which suffer from relatively small optical throughputs, known as the amount of spatial information provided per unit time[1,2], and limited imaging depth unable to extract signals from deep tissues
As with epifluorescence methods that suffer from a tradeoff between accuracy and scale, either a Gaussian or a Bessel light-sheet microscopy27–29 (LSM) system still has limited optical throughput, highlighting the difficulty for obtaining high spatial resolution across a very large FOV31
Summary
Rapid 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. To extract the various cellular profiles of different physiological functions within specimens, three-dimensional (3D) highresolution imaging is required throughout a mesoscale-sized volume Creating such a large-scale image dataset has posed significant challenges for conventional 3D light microscopy methods, which suffer from relatively small optical throughputs, known as the amount of spatial information provided per unit time[1,2], and limited imaging depth (up to a few hundred of microns) unable to extract signals from deep tissues. To minimize the effect from side-lobe excitation[30], Bessel LSM usually uses a high numerical-aperture (NA) detection objective with a small depth-offocus It is mainly optimized for the live imaging of organelles in a single or a few cells. A long acquisition time from several hours to days, as well as increased photobleaching to samples, still prevents the widespread applications of LSM to the highresolution mapping of entire mammalian organs/organisms
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