Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells
© 2020 The Authors. Expansion microscopy (ExM) is a technique that physically expands preserved cells and tissues before microscope imaging, so that conventional diffraction-limited microscopes can perform nanoscale-resolution imaging. In ExM, biomolecules or their markers are linked to a dense, swe...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2021
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| Online Access: | https://hdl.handle.net/1721.1/135960 |
| _version_ | 1811075806380687360 |
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| author | Zhang, Chi Kang, Jeong Seuk Asano, Shoh M Gao, Ruixuan Boyden, Edward S |
| author2 | Massachusetts Institute of Technology. Media Laboratory |
| author_facet | Massachusetts Institute of Technology. Media Laboratory Zhang, Chi Kang, Jeong Seuk Asano, Shoh M Gao, Ruixuan Boyden, Edward S |
| author_sort | Zhang, Chi |
| collection | MIT |
| description | © 2020 The Authors. Expansion microscopy (ExM) is a technique that physically expands preserved cells and tissues before microscope imaging, so that conventional diffraction-limited microscopes can perform nanoscale-resolution imaging. In ExM, biomolecules or their markers are linked to a dense, swellable gel network synthesized throughout a specimen. Mechanical homogenization of the sample (e.g., by protease digestion) and the addition of water enable isotropic swelling of the gel, so that the relative positions of biomolecules are preserved. We previously presented ExM protocols for analyzing proteins and RNAs in cells and tissues. Here we describe a cookbook-style ExM protocol for expanding cultured HeLa cells with immunostained microtubules, aimed to help newcomers familiarize themselves with the experimental setups and skills required to successfully perform ExM. Our aim is to help beginners, or students in a wet-lab classroom setting, learn all the key steps of ExM. © 2020 The Authors. |
| first_indexed | 2024-09-23T10:12:10Z |
| format | Article |
| id | mit-1721.1/135960 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T10:12:10Z |
| publishDate | 2021 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1359602023-09-28T19:51:01Z Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells Zhang, Chi Kang, Jeong Seuk Asano, Shoh M Gao, Ruixuan Boyden, Edward S Massachusetts Institute of Technology. Media Laboratory McGovern Institute for Brain Research at MIT Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Koch Institute for Integrative Cancer Research at MIT © 2020 The Authors. Expansion microscopy (ExM) is a technique that physically expands preserved cells and tissues before microscope imaging, so that conventional diffraction-limited microscopes can perform nanoscale-resolution imaging. In ExM, biomolecules or their markers are linked to a dense, swellable gel network synthesized throughout a specimen. Mechanical homogenization of the sample (e.g., by protease digestion) and the addition of water enable isotropic swelling of the gel, so that the relative positions of biomolecules are preserved. We previously presented ExM protocols for analyzing proteins and RNAs in cells and tissues. Here we describe a cookbook-style ExM protocol for expanding cultured HeLa cells with immunostained microtubules, aimed to help newcomers familiarize themselves with the experimental setups and skills required to successfully perform ExM. Our aim is to help beginners, or students in a wet-lab classroom setting, learn all the key steps of ExM. © 2020 The Authors. 2021-10-27T20:30:07Z 2021-10-27T20:30:07Z 2020 2021-03-23T18:46:22Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/135960 en 10.1002/CPNS.96 Current Protocols in Neuroscience Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Wiley Wiley |
| spellingShingle | Zhang, Chi Kang, Jeong Seuk Asano, Shoh M Gao, Ruixuan Boyden, Edward S Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title | Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title_full | Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title_fullStr | Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title_full_unstemmed | Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title_short | Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells |
| title_sort | expansion microscopy for beginners visualizing microtubules in expanded cultured hela cells |
| url | https://hdl.handle.net/1721.1/135960 |
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