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

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Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues Tay Won Shin , Hao Wang , Chi Zhang , Bobae An , Yangning Lu , Elizabeth Zhang , View ORCID Profile Xiaotang Lu , Emmanouil Karagiannis , Jeong Seuk Kang , Amauche Emenari , Panagiotis Symvoulidis , Shoh Asano , Leanne Lin , Emma Costa , IMAXT Grand Challenge Consortium , Adam H Marblestone , Narayanan Kasthuri , Li-Huei Tsai , Edward Boyden doi: https://doi.org/10.1101/2024.03.07.583776 Tay Won Shin 1 McGovern Institute, Massachusetts Institute of Technology, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Department of Media Arts and Sciences, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Hao Wang 2 McGovern Institute, Massachusetts Institute of Technology, Picower Inst. for Learning and Memory; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Chi Zhang 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Bobae An 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Yangning Lu 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Elizabeth Zhang 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Xiaotang Lu 4 Department of Cellular and Molecular Biology, Harvard University; Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Xiaotang Lu Emmanouil Karagiannis 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Jeong Seuk Kang 3 McGovern Institute, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Amauche Emenari 5 McGovern Institute, Massachusetts Institute of Technology, Department of Brain and Cognitive Sciences; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Panagiotis Symvoulidis 5 McGovern Institute, Massachusetts Institute of Technology, Department of Brain and Cognitive Sciences; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Shoh Asano 5 McGovern Institute, Massachusetts Institute of Technology, Department of Brain and Cognitive Sciences; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Leanne Lin 6 Department of Biological Engineering, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Emma Costa 7 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology; Find this author on Google Scholar Find this author on PubMed Search for this author on this site 8 -; Adam H Marblestone 9 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, present address: Convergent Research; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Narayanan Kasthuri 10 Center for Nanoscale Materials, Argonne National Laboratory, Department of Neurobiology, University of Chicago; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Li-Huei Tsai 11 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Picower Inst. for Learning and Memory; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Edward Boyden 12 McGovern Institute, Massachusetts Institute of Technology, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Howard Hughes Medical Institute, Department of Biological Engineering, Massachusetts Institute of Technology Find this author on Google Scholar Find this author on PubMed Search for this author on this site For correspondence: edboyden{at}mit.edu Abstract Info/History Metrics Supplementary material Preview PDF 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. Competing Interest Statement T.W.S. and E.S.B. are co-inventors on a patent application for umExM. E.S.B. is co-founder of a company seeking to deploy applications of ExM-related technologies. The other authors declare no competing interests. Footnotes The subject area has transitioned from bioengineering to neuroscience. Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . View the discussion thread. Back to top Previous Next Posted March 09, 2024. Download PDF Supplementary Material Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. 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Share Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues Tay Won Shin , Hao Wang , Chi Zhang , Bobae An , Yangning Lu , Elizabeth Zhang , Xiaotang Lu , Emmanouil Karagiannis , Jeong Seuk Kang , Amauche Emenari , Panagiotis Symvoulidis , Shoh Asano , Leanne Lin , Emma Costa , IMAXT Grand Challenge Consortium , Adam H Marblestone , Narayanan Kasthuri , Li-Huei Tsai , Edward Boyden bioRxiv 2024.03.07.583776; doi: https://doi.org/10.1101/2024.03.07.583776 Share This Article: Copy Citation Tools Dense, Continuous Membrane Labeling and Expansion Microscopy Visualization of Ultrastructure in Tissues Tay Won Shin , Hao Wang , Chi Zhang , Bobae An , Yangning Lu , Elizabeth Zhang , Xiaotang Lu , Emmanouil Karagiannis , Jeong Seuk Kang , Amauche Emenari , Panagiotis Symvoulidis , Shoh Asano , Leanne Lin , Emma Costa , IMAXT Grand Challenge Consortium , Adam H Marblestone , Narayanan Kasthuri , Li-Huei Tsai , Edward Boyden bioRxiv 2024.03.07.583776; doi: https://doi.org/10.1101/2024.03.07.583776 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Neuroscience Bioengineering Subject Areas All Articles Animal Behavior and Cognition (7646) Biochemistry (17728) Bioengineering (13917) Bioinformatics (42038) Biophysics (21489) Cancer Biology (18637) Cell Biology (25554) Clinical Trials (138) Developmental Biology (13403) Ecology (19941) Epidemiology (2067) Evolutionary Biology (24368) Genetics (15624) Genomics (22547) Immunology (17764) Microbiology (40475) Molecular Biology (17208) Neuroscience (88756) Paleontology (667) Pathology (2842) Pharmacology and Toxicology (4834) Physiology (7659) Plant Biology (15175) Scientific Communication and Education (2047) Synthetic Biology (4304) Systems Biology (9835) Zoology (2272)