Confocal Airy beam oblique light-sheet tomography for brain-wide cell type distribution and morphology.

Advanced brain-wide mapping is critical for addressing complex questions in neuroscience. However, current imaging methods are limited by throughput, resolution and signal-to-noise ratio, constraining their broader applicability. Here, we present confocal Airy beam integrated with single-photon oblique light-sheet tomography (CAB-OLST): a system that integrates single-photon excitation with a scanned Airy beam light sheet, virtual slit detection and automated mechanical sectioning. CAB-OLST enables high-throughput, high-resolution and high-signal-to-noise ratio volumetric imaging, achieving an optical resolution of 0.77 μm × 0.49 μm × 2.61 μm. This allows for mouse brain-wide cell type distribution mapping at a voxel size of 0.37 μm × 0.37 μm × 1.77 μm in 10 h and single-neuron projectome imaging with a voxel size of 0.26 μm × 0.26 μm × 1.06 μm over 58 h. Compared to existing light-sheet and point-scanning systems, CAB-OLST provides a scalable and robust platform for comprehensive neuronal morphology reconstruction and high-precision cell atlas generation.