@article {70161, title = {Mesoscale volumetric fluorescence imaging at nanoscale resolution by photochemical sectioning}, journal = {Science}, volume = {390}, year = {2025}, month = {2025 Oct 16}, pages = {eadr9109}, 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{\textendash}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

}, doi = {10.1126/science.adr9109}, url = {https://www.science.org/doi/abs/10.1126/science.adr9109}, author = {Wei Wang and Xiongtao Ruan and Gaoxiang Liu and Daniel E. Milkie and Wenping Li and Eric Betzig and Srigokul Upadhyayula and Ruixuan Gao} } @article {67849, title = {Rapid reconstruction of neural circuits using tissue expansion and lattice light sheet microscopy}, journal = {eLife}, year = {2022}, month = {2022 Oct 26}, abstract = {

Electron microscopy (EM) allows for the reconstruction of dense neuronal connectomes but suffers from low throughput, limiting its application to small numbers of reference specimens. We developed a protocol and analysis pipeline using tissue expansion and lattice light-sheet microscopy (ExLLSM) to rapidly reconstruct selected circuits across many samples with single synapse resolution and molecular contrast. We validate this approach in\ Drosophila, demonstrating that it yields synaptic counts similar to those obtained by EM, can be used to compare counts across sex and experience, and to correlate structural connectivity with functional connectivity. This approach fills a critical methodological gap in studying variability in the structure and function of neural circuits across individuals within and between species.

}, doi = {10.7554/eLife.81248}, url = {https://doi.org/10.7554/eLife.81248}, author = {Joshua L. Lillvis and Hideo Otsuna and Xiaoyu Ding and Igor Pisarev and Takashi Kawase and Jennifer Colonell and Konrad Rokicki and Cristian Goina and Ruixuan Gao and Amy Hu and Kaiyu Wang and John Bogovic and Daniel E. Milkie and Edward S. Boyden and Stephan Saalfeld and Paul W. Tillberg and Barry J. Dickson} }