Revealing nanostructures in brain tissue via protein decrowding by iterative expansion microscopy
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nihms-1838978.pdf
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Accepted version
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3.97 MB
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Author(s)
Sarkar, Deblina
Kang, Jinyoung
Wassie, Asmamaw T
Schroeder, Margaret E
Peng, Zhuyu
Tarr, Tyler B
Tang, Ai-Hui
Niederst, Emily D
Young, Jennie Z
Su, Hanquan
Date Issued
2022
Journal
Nature Biomedical Engineering
Publisher
Springer Science and Business Media LLC
Citation
Sarkar, Deblina, Kang, Jinyoung, Wassie, Asmamaw T, Schroeder, Margaret E, Peng, Zhuyu et al. 2022. "Revealing nanostructures in brain tissue via protein decrowding by iterative expansion microscopy." Nature Biomedical Engineering, 6 (9).
Version
Author's final manuscript
Abstract
Many crowded biomolecular structures in cells and tissues are inaccessible to labelling antibodies. To understand how proteins within these structures are arranged with nanoscale precision therefore requires that these structures be decrowded before labelling. Here we show that an iterative variant of expansion microscopy (the permeation of cells and tissues by a swellable hydrogel followed by isotropic hydrogel expansion, to allow for enhanced imaging resolution with ordinary microscopes) enables the imaging of nanostructures in expanded yet otherwise intact tissues at a resolution of about 20 nm. The method, which we named 'expansion revealing' and validated with DNA-probe-based super-resolution microscopy, involves gel-anchoring reagents and the embedding, expansion and re-embedding of the sample in homogeneous swellable hydrogels. Expansion revealing enabled us to use confocal microscopy to image the alignment of pre-synaptic calcium channels with post-synaptic scaffolding proteins in intact brain circuits, and to uncover periodic amyloid nanoclusters containing ion-channel proteins in brain tissue from a mouse model of Alzheimer's disease. Expansion revealing will enable the further discovery of previously unseen nanostructures within cells and tissues.
MIT Department
Massachusetts Institute of Technology. Media Laboratory
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Picower Institute for Learning and Memory
Koch Institute for Integrative Cancer Research at MIT
Howard Hughes Medical Institute
Program in Media Arts and Sciences (Massachusetts Institute of Technology)
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DOI of Published Version
https://doi.org/10.1038/S41551-022-00912-3
