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Main Menu - Block
- Overview
- Anatomy and Histology
- Cryo-Electron Microscopy
- Electron Microscopy
- Flow Cytometry
- Gene Targeting and Transgenics
- High Performance Computing
- Immortalized Cell Line Culture
- Integrative Imaging
- Invertebrate Shared Resource
- Janelia Experimental Technology
- Mass Spectrometry
- Media Prep
- Molecular Genomics
- Primary & iPS Cell Culture
- Project Pipeline Support
- Project Technical Resources
- Quantitative Genomics
- Scientific Computing
- Viral Tools
- Vivarium
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 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 a photochemical strategy for spatially precise sectioning of specimens. By combining serial photochemical sectioning with lattice light-sheet imaging and petabyte-scale computation, 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. High-resolution microscopes have a short working distance, making it difficult to see deep within large biological samples such as an intact brain. Slicing the tissue with a blade can reach deeper, but this often distorts or destroys the fine structures that scientists want to study. By embedding a sample in a light-sensitive hydrogel, Wang et al. demonstrated a gentler approach using a precise ray or sheet of light to dissolve or cut away tissue layer by layer. After each layer is removed, the newly exposed surface is imaged, allowing for a complete, high-resolution, three-dimensional reconstruction without damaging physical contact.
bioRxiv preprint: https://www.biorxiv.org/content/10.1101/2024.08.01.605857v1
