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Nat. Commun., Published: 12 February 2025 | https://doi.org/10.1038/s41467-025-56641-z
Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues
Tay Won Shin1,2,3, Hao Wang1,4,16, Chi Zhang1,16, Bobae An1, Yangning Lu1, Elizabeth Zhang1, Xiaotang Lu5, Emmanouil D. Karagiannis1, Jeong Seuk Kang1, Amauche Emenari1,2, Panagiotis Symvoulidis1, Shoh Asano1,13, Leanne Lin6, Emma K. Costa2,14, IMAXT Grand Challenge Consortium*, Adam H. Marblestone1,15, Narayanan Kasthuri7,8, Li-Huei Tsai2,4 & Edward S. Boyden1,2,3,6,9,10,11,12
1McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
2Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
3Department of Media Arts and Sciences, Massachusetts Institute of Technology, Cambridge, MA 02140, USA.
4Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
5Department of Cellular and Molecular Biology, Harvard University, Cambridge, MA 02138, USA.
6Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
7Center for Nanoscale Materials, Argonne National Laboratory, Lemont, IL 60439, USA.
8Department of Neurobiology, University of Chicago, hicago, IL 60637, USA.
9Koch Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
10Center for Neurobiological Engineering, assachusetts Institute of Technology, Cambridge, MA 02139, USA.
11K. Lisa Yang Center for Bionics, Cambridge, MA 02139, USA.
12Howard Hughes Medical nstitute, Cambridge, MA 02139, USA.
13Present address: Pfizer Inc, Cambridge, MA 02139, USA.
14Present address: Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
15Present address: Convergent Research, Cambridge, MA 02140, USA.
16These authors contributed equally: Hao Wang, Chi Zhang.
Corresponding author
Correspondence to Edward S. Boyden.
1McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
2Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
3Department of Media Arts and Sciences, Massachusetts Institute of Technology, Cambridge, MA 02140, USA.
4Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
5Department of Cellular and Molecular Biology, Harvard University, Cambridge, MA 02138, USA.
6Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
7Center for Nanoscale Materials, Argonne National Laboratory, Lemont, IL 60439, USA.
8Department of Neurobiology, University of Chicago, hicago, IL 60637, USA.
9Koch Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
10Center for Neurobiological Engineering, assachusetts Institute of Technology, Cambridge, MA 02139, USA.
11K. Lisa Yang Center for Bionics, Cambridge, MA 02139, USA.
12Howard Hughes Medical nstitute, Cambridge, MA 02139, USA.
13Present address: Pfizer Inc, Cambridge, MA 02139, USA.
14Present address: Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
15Present address: Convergent Research, Cambridge, MA 02140, USA.
16These authors contributed equally: Hao Wang, Chi Zhang.
Corresponding author
Correspondence to Edward S. Boyden.
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.
논문정보
- Research article
- 2025년 02월 (BRIC 등록일 2025-02-13)
- 국외 연구진
- 바이오·의료융합 > 바이오소재
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