Visualizing Drosophila connectome with multiview light-sheet macrophotography and iterative expansion microscopy

We present a novel integration of iterative expansion microscopy (ExM) and light-sheet illuminated macro-photography, enabling rapid 3D imaging of large biological samples with unprecedented resolution. This method achieves over 50-fold sample expansion, providing electron microscopy-level (~10 nm) and optical microscopy-level (~500 nm) resolution. The platform ensures high isotropic spatial resolution, exceptional optical throughput, and rapid volume mapping by leveraging multi-view imaging and advanced CMOS sensors. Demonstrated on Drosophila brains and whole-body specimens, this approach opens new frontiers for large-scale connectomics and high-resolution brain mapping.