Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs.
Expansion microscopy (ExM), by physically enlarging specimens in an isotropic fashion, enables nanoimaging on standard light microscopes. Key to existing ExM protocols is the equipping of different kinds of molecules, with different kinds of anchoring moieties, so they can all be pulled apart from e...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0291506 |
| _version_ | 1797674060077858816 |
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| author | Yi Cui Gaojie Yang Daniel R Goodwin Ciara H O'Flanagan Anubhav Sinha Chi Zhang Kristina E Kitko Tay Won Shin Demian Park Samuel Aparicio CRUK IMAXT Grand Challenge Consortium Edward S Boyden |
| author_facet | Yi Cui Gaojie Yang Daniel R Goodwin Ciara H O'Flanagan Anubhav Sinha Chi Zhang Kristina E Kitko Tay Won Shin Demian Park Samuel Aparicio CRUK IMAXT Grand Challenge Consortium Edward S Boyden |
| author_sort | Yi Cui |
| collection | DOAJ |
| description | Expansion microscopy (ExM), by physically enlarging specimens in an isotropic fashion, enables nanoimaging on standard light microscopes. Key to existing ExM protocols is the equipping of different kinds of molecules, with different kinds of anchoring moieties, so they can all be pulled apart from each other by polymer swelling. Here we present a multifunctional anchor, an acrylate epoxide, that enables proteins and RNAs to be equipped with anchors in a single experimental step. This reagent simplifies ExM protocols and reduces cost (by 2-10-fold for a typical multiplexed ExM experiment) compared to previous strategies for equipping RNAs with anchors. We show that this united ExM (uniExM) protocol can be used to preserve and visualize RNA transcripts, proteins in biologically relevant ultrastructures, and sets of RNA transcripts in patient-derived xenograft (PDX) cancer tissues and may support the visualization of other kinds of biomolecular species as well. uniExM may find many uses in the simple, multimodal nanoscale analysis of cells and tissues. |
| first_indexed | 2024-03-11T21:53:37Z |
| format | Article |
| id | doaj.art-51e6dcd0c8df44839fd1fba616c089e7 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-03-11T21:53:37Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-51e6dcd0c8df44839fd1fba616c089e72023-09-26T05:31:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01189e029150610.1371/journal.pone.0291506Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs.Yi CuiGaojie YangDaniel R GoodwinCiara H O'FlanaganAnubhav SinhaChi ZhangKristina E KitkoTay Won ShinDemian ParkSamuel AparicioCRUK IMAXT Grand Challenge ConsortiumEdward S BoydenExpansion microscopy (ExM), by physically enlarging specimens in an isotropic fashion, enables nanoimaging on standard light microscopes. Key to existing ExM protocols is the equipping of different kinds of molecules, with different kinds of anchoring moieties, so they can all be pulled apart from each other by polymer swelling. Here we present a multifunctional anchor, an acrylate epoxide, that enables proteins and RNAs to be equipped with anchors in a single experimental step. This reagent simplifies ExM protocols and reduces cost (by 2-10-fold for a typical multiplexed ExM experiment) compared to previous strategies for equipping RNAs with anchors. We show that this united ExM (uniExM) protocol can be used to preserve and visualize RNA transcripts, proteins in biologically relevant ultrastructures, and sets of RNA transcripts in patient-derived xenograft (PDX) cancer tissues and may support the visualization of other kinds of biomolecular species as well. uniExM may find many uses in the simple, multimodal nanoscale analysis of cells and tissues.https://doi.org/10.1371/journal.pone.0291506 |
| spellingShingle | Yi Cui Gaojie Yang Daniel R Goodwin Ciara H O'Flanagan Anubhav Sinha Chi Zhang Kristina E Kitko Tay Won Shin Demian Park Samuel Aparicio CRUK IMAXT Grand Challenge Consortium Edward S Boyden Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. PLoS ONE |
| title | Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. |
| title_full | Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. |
| title_fullStr | Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. |
| title_full_unstemmed | Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. |
| title_short | Expansion microscopy using a single anchor molecule for high-yield multiplexed imaging of proteins and RNAs. |
| title_sort | expansion microscopy using a single anchor molecule for high yield multiplexed imaging of proteins and rnas |
| url | https://doi.org/10.1371/journal.pone.0291506 |
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