Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues
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), whi...
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Nature Portfolio
2025-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56641-z |
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| author | Tay Won Shin Hao Wang Chi Zhang Bobae An Yangning Lu Elizabeth Zhang Xiaotang Lu Emmanouil D. Karagiannis Jeong Seuk Kang Amauche Emenari Panagiotis Symvoulidis Shoh Asano Leanne Lin Emma K. Costa IMAXT Grand Challenge Consortium Adam H. Marblestone Narayanan Kasthuri Li-Huei Tsai Edward S. Boyden |
| author_facet | Tay Won Shin Hao Wang Chi Zhang Bobae An Yangning Lu Elizabeth Zhang Xiaotang Lu Emmanouil D. Karagiannis Jeong Seuk Kang Amauche Emenari Panagiotis Symvoulidis Shoh Asano Leanne Lin Emma K. Costa IMAXT Grand Challenge Consortium Adam H. Marblestone Narayanan Kasthuri Li-Huei Tsai Edward S. Boyden |
| author_sort | Tay Won Shin |
| collection | DOAJ |
| description | 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 an innovative membrane label and optimized expansion microscopy protocol, to support dense labeling of membranes in tissues for nanoscale visualization. We validate the high signal-to-background ratio, and uniformity and continuity, of umExM membrane labeling in brain slices, which supports the imaging of membranes and proteins at a resolution of ~60 nm on a confocal microscope. We demonstrate 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, yields ~35 nm resolution imaging, pointing towards the potential for electron microscopy resolution visualization of brain membranes on ordinary light microscopes. |
| format | Article |
| id | doaj-art-670f2abb61e14bc2863da95da7a74b58 |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-670f2abb61e14bc2863da95da7a74b582025-08-20T03:00:58ZengNature PortfolioNature Communications2041-17232025-02-0116111710.1038/s41467-025-56641-zDense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissuesTay Won Shin0Hao Wang1Chi Zhang2Bobae An3Yangning Lu4Elizabeth Zhang5Xiaotang Lu6Emmanouil D. Karagiannis7Jeong Seuk Kang8Amauche Emenari9Panagiotis Symvoulidis10Shoh Asano11Leanne Lin12Emma K. Costa13IMAXT Grand Challenge ConsortiumAdam H. Marblestone14Narayanan Kasthuri15Li-Huei Tsai16Edward S. Boyden17McGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyDepartment of Cellular and Molecular Biology, Harvard UniversityMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Brain and Cognitive Sciences, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyCenter for Nanoscale Materials, Argonne National LaboratoryDepartment of Brain and Cognitive Sciences, Massachusetts Institute of TechnologyMcGovern Institute, Massachusetts Institute of TechnologyAbstract 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 an innovative membrane label and optimized expansion microscopy protocol, to support dense labeling of membranes in tissues for nanoscale visualization. We validate the high signal-to-background ratio, and uniformity and continuity, of umExM membrane labeling in brain slices, which supports the imaging of membranes and proteins at a resolution of ~60 nm on a confocal microscope. We demonstrate 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, yields ~35 nm resolution imaging, pointing towards the potential for electron microscopy resolution visualization of brain membranes on ordinary light microscopes.https://doi.org/10.1038/s41467-025-56641-z |
| spellingShingle | Tay Won Shin Hao Wang Chi Zhang Bobae An Yangning Lu Elizabeth Zhang Xiaotang Lu Emmanouil D. Karagiannis Jeong Seuk Kang Amauche Emenari Panagiotis Symvoulidis Shoh Asano Leanne Lin Emma K. Costa IMAXT Grand Challenge Consortium Adam H. Marblestone Narayanan Kasthuri Li-Huei Tsai Edward S. Boyden Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues Nature Communications |
| title | Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| title_full | Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| title_fullStr | Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| title_full_unstemmed | Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| title_short | Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| title_sort | dense continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues |
| url | https://doi.org/10.1038/s41467-025-56641-z |
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