Curved light sheet microscopy for centimetre-scale cleared tissue imaging

Imaging large cleared tissues requires scaling the throughput of imaging techniques. Light sheet microscopy is a promising technique for high-throughput imaging; however, its reliance on conventional microscope objectives limits the optimization of the trade-off between spatial resolution and field of view. Here we introduce curved light sheet microscope to perform optical sectioning with curved light sheets. This concept addresses the long-standing field curvature problem and lowers the barriers in designing high-throughput objectives. Leveraging a customized objective, the curved light sheet microscope achieves diffraction-limited resolution of 1.0 μm laterally and 2.5 μm axially, with uniform contrast over a field of view of more than 1 × 1 cm2. Our technique is also compatible with various tissue clearing techniques. We demonstrate that imaging an entire intact cleared mouse brain at a voxel size of 0.625 × 0.625 × 1.25 μm3 can be completed in less than 3 h, without the need for image tiling. We share a full optical description of the objective and report imaging of neuronal and vascular networks, as well as tracing of brain-wide long-distance axonal projections in intact mouse brains. Light sheet microscopy with curved light sheets enables tiling-free imaging of an entire intact cleared mouse brain with lateral and axial spatial resolutions of 1.0 μm and 2.5 μm, respectively, in less than 3 h.