Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
<jats:title>Abstract</jats:title><jats:p>The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here,...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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Springer Science and Business Media LLC
2021
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| Online Access: | https://hdl.handle.net/1721.1/138174 |
| _version_ | 1826207459604168704 |
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| author | Lee, Hsuan Yu, Chih-Chieh Boyden, Edward S Zhuang, Xiaowei Kosuri, Pallav |
| author2 | McGovern Institute for Brain Research at MIT |
| author_facet | McGovern Institute for Brain Research at MIT Lee, Hsuan Yu, Chih-Chieh Boyden, Edward S Zhuang, Xiaowei Kosuri, Pallav |
| author_sort | Lee, Hsuan |
| collection | MIT |
| description | <jats:title>Abstract</jats:title><jats:p>The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here, we quantified the accuracy of gel embedding by using stochastic optical reconstruction microscopy (STORM) to image DNA origami with well-defined structures. We found that embedding in hydrogels based on polyacrylamide, the most widely used chemistry in ExM, resulted in random displacements of labeled sites with a standard deviation of ~ 16 nm. In contrast, we found that embedding in tetra-gel, a hydrogel that does not depend on free-radical chain-growth polymerization, preserved labeled sites with a standard deviation of less than 5 nm. By combining tetra-gel ExM with STORM, we were able to resolve 11-nm structural features without the loss in accuracy seen with polyacrylamide gels. Our study thus provides direct measurements of the single-molecule distortions resulting from hydrogel embedding, and presents a way to improve super-resolution microscopy through combination with tetra-gel ExM.</jats:p> |
| first_indexed | 2024-09-23T13:49:59Z |
| format | Article |
| id | mit-1721.1/138174 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:49:59Z |
| publishDate | 2021 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1381742023-02-03T20:38:21Z Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy Lee, Hsuan Yu, Chih-Chieh Boyden, Edward S Zhuang, Xiaowei Kosuri, Pallav McGovern Institute for Brain Research at MIT Howard Hughes Medical Institute Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences <jats:title>Abstract</jats:title><jats:p>The accuracy of expansion microscopy (ExM) depends on the structural preservation of samples embedded in a hydrogel. However, it has been unknown to what extent gel embedding alters the molecular positions of individual labeled sites. Here, we quantified the accuracy of gel embedding by using stochastic optical reconstruction microscopy (STORM) to image DNA origami with well-defined structures. We found that embedding in hydrogels based on polyacrylamide, the most widely used chemistry in ExM, resulted in random displacements of labeled sites with a standard deviation of ~ 16 nm. In contrast, we found that embedding in tetra-gel, a hydrogel that does not depend on free-radical chain-growth polymerization, preserved labeled sites with a standard deviation of less than 5 nm. By combining tetra-gel ExM with STORM, we were able to resolve 11-nm structural features without the loss in accuracy seen with polyacrylamide gels. Our study thus provides direct measurements of the single-molecule distortions resulting from hydrogel embedding, and presents a way to improve super-resolution microscopy through combination with tetra-gel ExM.</jats:p> 2021-11-19T20:06:26Z 2021-11-19T20:06:26Z 2021-12 2021-11-19T20:02:31Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/138174 Lee, Hsuan, Yu, Chih-Chieh, Boyden, Edward S, Zhuang, Xiaowei and Kosuri, Pallav. 2021. "Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy." Scientific Reports, 11 (1). en 10.1038/s41598-021-96258-y Scientific Reports Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Scientific Reports |
| spellingShingle | Lee, Hsuan Yu, Chih-Chieh Boyden, Edward S Zhuang, Xiaowei Kosuri, Pallav Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title | Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title_full | Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title_fullStr | Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title_full_unstemmed | Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title_short | Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy |
| title_sort | tetra gel enables superior accuracy in combined super resolution imaging and expansion microscopy |
| url | https://hdl.handle.net/1721.1/138174 |
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