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Decoding large neural networks using tissue clearing, tissue expansion, and tiling light sheet microscopy techniques

The combination of tissue clearing, tissue expansion, and light sheet microscopy techniques offers a solution to decode large neural networks. We first developed a tiling light sheet microscope, compatible with all tissue clearing methods, for rapid multicolor 3D imaging of cleared tissues at multiple resolution scales. The resolving ability is further improved to sub-100 nm via tissue expansion. The microscope uses tiling light sheets to achieve higher spatial resolution and better optical sectioning ability than conventional light sheet microscopes. The illumination light is phase modulated to optimize the imaging performance, keep the microscope aligned, ensure accurate multicolor 3D imaging and make the microscope reliable and easy to operate. We further show the use of our method to understand the function, development, and regeneration of the mouse spinal cord locomotor neural network and the planarian neural muscular network. We show our method could advance the understanding of large neural networks significantly.

19 2023-12-25

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