Abstract:

Optical imaging has made tremendous inroads into revealing cellular and subcellular structures with molecular contrast in brain tissue. However, existing high-resolution fluorescence microscopes are physically limited by objective-to-specimen distance, which prevents the study of whole-mount specimens without physical sectioning. To address this challenge, we developed VIPS (volumetric imaging by photochemical sectioning), a technique integrating advanced lattice light sheet expansion microscopy for 3D super-resolution and a novel photochemical strategy for spatially precise sectioning of specimens. With this method, we imaged and reconstructed axons and myelin 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 brain, highlighting the potential for peta- to exabyte-scale super-resolution studies using this approach. This technology holds the promise to establish the foundation for an optical pipeline to deliver synaptic-level dense connectomes of whole brains.

Profile

Advanced Bioimaging Center, Department of Molecular and Cell Biology, University of California, Berkeley