Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy
Abstract 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 usi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-08-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-96258-y |
| _version_ | 1818856633137102848 |
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| author | Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri |
| author_facet | Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri |
| author_sort | Hsuan Lee |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-19T08:27:36Z |
| format | Article |
| id | doaj.art-07fd9716e4d444bcb23396fa05e84b9c |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T08:27:36Z |
| publishDate | 2021-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-07fd9716e4d444bcb23396fa05e84b9c2022-12-21T20:29:15ZengNature PortfolioScientific Reports2045-23222021-08-011111710.1038/s41598-021-96258-yTetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopyHsuan Lee0Chih-Chieh Yu1Edward S. Boyden2Xiaowei Zhuang3Pallav Kosuri4Howard Hughes Medical Institute, Harvard UniversityMcGovern Institute for Brain Research, MITMcGovern Institute for Brain Research, MITHoward Hughes Medical Institute, Harvard UniversityHoward Hughes Medical Institute, Harvard UniversityAbstract 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.https://doi.org/10.1038/s41598-021-96258-y |
| spellingShingle | Hsuan Lee Chih-Chieh Yu Edward S. Boyden Xiaowei Zhuang Pallav Kosuri Tetra-gel enables superior accuracy in combined super-resolution imaging and expansion microscopy Scientific Reports |
| 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://doi.org/10.1038/s41598-021-96258-y |
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