Nanoscale volumetric fluorescence imaging via photochemical sectioning

Wang, W.; Ruan, X.; Liu, G.; Milkie, D. E.; Li, W.; Betzig, E.; Upadhyayula, S.; Gao, R.

doi:10.1101/2024.08.01.605857

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bioRxiv · 10.1101/2024.08.01.605857

Nanoscale volumetric fluorescence imaging via photochemical sectioning

Wang, W.·Ruan, X.·Liu, G.·Milkie, D. E.·Li, W.·Betzig, E.·Upadhyayula, S.·Gao, R.

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Abstract

Optical nanoscopy of intact biological specimens has been transformed by recent advancements in hydrogel-based tissue clearing and expansion, enabling the imaging of cellular and subcellular structures with molecular contrast. However, existing high-resolution fluorescence microscopes have limited imaging depth, which prevents the study of whole-mount specimens without physical sectioning. To address this challenge, we developed "photochemical sectioning," a spatially precise, light-based sample sectioning process. By combining photochemical sectioning with volumetric lattice light-sheet imaging and petabyte-scale computation, we imaged and reconstructed axons and myelination sheaths across entire mouse olfactory bulbs at nanoscale resolution. An olfactory-bulb-wide analysis of myelinated and unmyelinated axons revealed distinctive patterns of axon degeneration and de-/dysmyelination in the neurodegenerative mouse, highlighting the potential for peta- to exabyte-scale super-resolution studies using this approach.

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Wang, W., Ruan, X., Liu, G., Milkie, D. E., Li, W., Betzig, E., Upadhyayula, S., Gao, R.. 2024-08-05. Nanoscale volumetric fluorescence imaging via photochemical sectioning. https://doi.org/10.1101/2024.08.01.605857

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