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Benefits and advantages of Light-Sheet Microscopy for fast, high-resolution tissue imaging - Hybrid light sheet geometry for wide area scouting and high resolution zoom The hybrid microscope architecture consists of three objectives positioned below the specimen. One objective is used for light-sheet illumination, and the other two objectives are used for orthogonal dual-objective (ODO) and non-orthogonal dual-objective (NODO) imaging. By using a motorized stage, tiled imaging is possible with both paths over a large 12 × 7.5 × 1 cm (XYZ) imaging volume, accommodating multiple intact cleared organs and large tissue slabs mounted in an array of specimen holders. Nature Methods 2022 https://images.squarespace-cdn.com/content/v1/61bb686d64a5d61acdaa3b10/a3fa28e0-74a5-4d01-bc98-d5471a7c0bae/Scout-Zoom+HOTLS Blog - Benefits and advantages of Light-Sheet Microscopy for fast, high-resolution tissue imaging - Comparing Scout and Zoom resolution i, Representative ODO imaging results of an entire intact CUBIC-cleared mouse brain with arterial (αSMA) and nuclear (SYTOX-G) staining. The ODO imaging path is able to clearly resolve vasculature in both the xy and xz planes (insets). The size and direction of tiling is annotated. j,k, Targeted NODO imaging of a sub-region centered on a branching arteriole resolves individual smooth muscle cells and sub-nuclear features that are not resolved by ODO imaging. Scale-bar lengths are as follows: i, 1mm (insets, 500 μm); j, 100 μm; and k, 10 μm. 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