Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies
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Boyden_Protein-retention.pdf
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Author(s)
English, Brian P
Gao, Linyi
Suk, Ho-Jun
Yoshida, Fumiaki
DeGennaro, Ellen M
Roossien, Douglas H
Cai, Dawen
Tillberg, Paul W.
Chen, Fei
Piatkevich, Kiryl
Date Issued
July 2016
Journal
Nature Biotechnology
Publisher
Nature Publishing Group
Citation
Tillberg, Paul W; Chen, Fei; Piatkevich, Kiryl D; Zhao, Yongxin; Yu, Chih-Chieh (Jay); English, Brian P; Gao, Linyi et al. “Protein-Retention Expansion Microscopy of Cells and Tissues Labeled Using Standard Fluorescent Proteins and Antibodies.” Nature Biotechnology 34, no. 9 (July 2016): 987–992. © 2016 Macmillan Publishers Limited, part of Springer Nature
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Author's final manuscript
Abstract
Expansion microscopy (ExM) enables imaging of preserved specimens with nanoscale precision on diffraction-limited instead of specialized super-resolution microscopes. ExM works by physically separating fluorescent probes after anchoring them to a swellable gel. The first ExM method did not result in the retention of native proteins in the gel and relied on custom-made reagents that are not widely available. Here we describe protein retention ExM (proExM), a variant of ExM in which proteins are anchored to the swellable gel, allowing the use of conventional fluorescently labeled antibodies and streptavidin, and fluorescent proteins. We validated and demonstrated the utility of proExM for multicolor super-resolution (~70 nm) imaging of cells and mammalian tissues on conventional microscopes.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Media Laboratory
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/nbt.3625
