2. Academic Validation
  3. Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues

Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues

  • bioRxiv. 2024 Mar 8:2024.03.07.583776. doi: 10.1101/2024.03.07.583776.
Tay Won Shin 1 2 3,  Hao Wang 1 4,  Chi Zhang 1,  Bobae An 1,  Yangning Lu 1,  Elizabeth Zhang 1,  Xiaotang Lu 5,  Emmanouil D Karagiannis 1,  Jeong Seuk Kang 1,  Amauche Emenari 1 2,  Panagiotis Symvoulidis 1 2,  Shoh Asano 1,  Leanne Lin 6,  Emma K Costa 2,  IMAXT Grand Challenge Consortium,  Adam H Marblestone 1 7,  Narayanan Kasthuri 8 9,  Li-Huei Tsai 2 4,  Edward S Boyden 1 2 10 6 11 12 13 3
Affiliations

Affiliations

  • 1 McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 2 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 3 Department of Media Arts and Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 4 Picower Inst. for Learning and Memory, Cambridge.
  • 5 Department of Cellular and Molecular Biology, Harvard University, Cambridge, MA, United States.
  • 6 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 7 present address: Convergent Research.
  • 8 Center for Nanoscale Materials, Argonne National Laboratory, Lemont, IL, USA.
  • 9 Department of Neurobiology, University of Chicago, Chicago, IL, USA.
  • 10 Howard Hughes Medical Institute, Cambridge, MA 02139.
  • 11 Koch Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 12 Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 13 K. Lisa Yang Center for Bionics, Massachusetts Institute of Technology, Cambridge, MA 02139.
PMID: 38496681 DOI: 10.1101/2024.03.07.583776
Abstract

Lipid membranes are key to the nanoscale compartmentalization of biological systems, but fluorescent visualization of them in intact tissues, with nanoscale precision, is challenging to do with high labeling density. Here, we report ultrastructural membrane expansion microscopy (umExM), which combines a novel membrane label and optimized expansion microscopy protocol, to support dense labeling of membranes in tissues for nanoscale visualization. We validated the high signal-to-background ratio, and uniformity and continuity, of umExM membrane labeling in brain slices, which supported the imaging of membranes and proteins at a resolution of ~60 nm on a confocal microscope. We demonstrated the utility of umExM for the segmentation and tracing of neuronal processes, such as axons, in mouse brain tissue. Combining umExM with optical fluctuation imaging, or iterating the expansion process, yielded ~35 nm resolution imaging, pointing towards the potential for electron microscopy resolution visualization of brain membranes on ordinary light microscopes.

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