Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies
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...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2017
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| Online Access: | http://hdl.handle.net/1721.1/108514 https://orcid.org/0000-0003-0254-4741 https://orcid.org/0000-0003-3776-4605 https://orcid.org/0000-0003-4188-5725 https://orcid.org/0000-0001-8713-0446 https://orcid.org/0000-0002-3579-0327 https://orcid.org/0000-0002-2206-2590 https://orcid.org/0000-0002-0899-6709 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0002-5938-4227 https://orcid.org/0000-0002-0419-3351 |
| _version_ | 1826203913464840192 |
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| author | 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 Zhao, Yongxin Yu, Chih-Chieh Martorell, Anthony Gong, Guanyu Seneviratne, Uthpala Indrajith Tannenbaum, Steven R Desimone, Robert Boyden, Edward |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering 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 Zhao, Yongxin Yu, Chih-Chieh Martorell, Anthony Gong, Guanyu Seneviratne, Uthpala Indrajith Tannenbaum, Steven R Desimone, Robert Boyden, Edward |
| author_sort | English, Brian P |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T12:45:20Z |
| format | Article |
| id | mit-1721.1/108514 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:45:20Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1085142022-10-01T10:56:04Z Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies 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 Zhao, Yongxin Yu, Chih-Chieh Martorell, Anthony Gong, Guanyu Seneviratne, Uthpala Indrajith Tannenbaum, Steven R Desimone, Robert Boyden, Edward 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 Tillberg, Paul W. Chen, Fei Piatkevich, Kiryl Zhao, Yongxin Yu, Chih-Chieh Gao, Linyi Martorell, Anthony Suk, Ho-Jun Gong, Guanyu Seneviratne, Uthpala Indrajith Tannenbaum, Steven R Desimone, Robert Boyden, Edward 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. United States. National Institutes of Health (1R01GM104948) United States. National Institutes of Health (1DP1NS087724) United States. National Institutes of Health ( NIH 1R01EY023173) United States. National Institutes of Health (1U01MH106011) 2017-04-28T20:30:04Z 2017-04-28T20:30:04Z 2016-07 2015-11 Article http://purl.org/eprint/type/JournalArticle 1087-0156 1546-1696 http://hdl.handle.net/1721.1/108514 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 https://orcid.org/0000-0003-0254-4741 https://orcid.org/0000-0003-3776-4605 https://orcid.org/0000-0003-4188-5725 https://orcid.org/0000-0001-8713-0446 https://orcid.org/0000-0002-3579-0327 https://orcid.org/0000-0002-2206-2590 https://orcid.org/0000-0002-0899-6709 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0002-5938-4227 https://orcid.org/0000-0002-0419-3351 en_US http://dx.doi.org/10.1038/nbt.3625 Nature Biotechnology 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. application/pdf Nature Publishing Group PMC |
| spellingShingle | 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 Zhao, Yongxin Yu, Chih-Chieh Martorell, Anthony Gong, Guanyu Seneviratne, Uthpala Indrajith Tannenbaum, Steven R Desimone, Robert Boyden, Edward Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title | Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title_full | Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title_fullStr | Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title_full_unstemmed | Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title_short | Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| title_sort | protein retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies |
| url | http://hdl.handle.net/1721.1/108514 https://orcid.org/0000-0003-0254-4741 https://orcid.org/0000-0003-3776-4605 https://orcid.org/0000-0003-4188-5725 https://orcid.org/0000-0001-8713-0446 https://orcid.org/0000-0002-3579-0327 https://orcid.org/0000-0002-2206-2590 https://orcid.org/0000-0002-0899-6709 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0002-5938-4227 https://orcid.org/0000-0002-0419-3351 |
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