Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues
Expansion microscopy (ExM) is a recently developed technique that enables nanoscale-resolution imaging of preserved cells and tissues on conventional diffraction-limited microscopes via isotropic physical expansion of the specimens before imaging. In ExM, biomolecules and/or fluorescent labels in th...
| Principais autores: | , , , , , |
|---|---|
| Outros Autores: | |
| Formato: | Artigo |
| Idioma: | English |
| Publicado em: |
Wiley
2020
|
| Acesso em linha: | https://hdl.handle.net/1721.1/125343 |
| _version_ | 1826188617523920896 |
|---|---|
| author | Asano, Shoh M. Gao, Ruixuan Wassie, Asmamaw T. Tillberg, Paul W. Chen, Fei Boyden, Edward |
| author2 | McGovern Institute for Brain Research at MIT |
| author_facet | McGovern Institute for Brain Research at MIT Asano, Shoh M. Gao, Ruixuan Wassie, Asmamaw T. Tillberg, Paul W. Chen, Fei Boyden, Edward |
| author_sort | Asano, Shoh M. |
| collection | MIT |
| description | Expansion microscopy (ExM) is a recently developed technique that enables nanoscale-resolution imaging of preserved cells and tissues on conventional diffraction-limited microscopes via isotropic physical expansion of the specimens before imaging. In ExM, biomolecules and/or fluorescent labels in the specimen are linked to a dense, expandable polymer matrix synthesized evenly throughout the specimen, which undergoes 3-dimensional expansion by ∼4.5 fold linearly when immersed in water. Since our first report, versions of ExM optimized for visualization of proteins, RNA, and other biomolecules have emerged. Here we describe best-practice, step-by-step ExM protocols for performing analysis of proteins (protein retention ExM, or proExM) as well as RNAs (expansion fluorescence in situ hybridization, or ExFISH), using chemicals and hardware found in a typical biology lab. Furthermore, a detailed protocol for handling and mounting expanded samples and for imaging them with confocal and light-sheet microscopes is provided. |
| first_indexed | 2024-09-23T08:02:28Z |
| format | Article |
| id | mit-1721.1/125343 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:02:28Z |
| publishDate | 2020 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1253432022-09-30T01:54:51Z Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues Asano, Shoh M. Gao, Ruixuan Wassie, Asmamaw T. Tillberg, Paul W. Chen, Fei Boyden, Edward McGovern Institute for Brain Research at MIT Massachusetts Institute of Technology. Media Laboratory Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Expansion microscopy (ExM) is a recently developed technique that enables nanoscale-resolution imaging of preserved cells and tissues on conventional diffraction-limited microscopes via isotropic physical expansion of the specimens before imaging. In ExM, biomolecules and/or fluorescent labels in the specimen are linked to a dense, expandable polymer matrix synthesized evenly throughout the specimen, which undergoes 3-dimensional expansion by ∼4.5 fold linearly when immersed in water. Since our first report, versions of ExM optimized for visualization of proteins, RNA, and other biomolecules have emerged. Here we describe best-practice, step-by-step ExM protocols for performing analysis of proteins (protein retention ExM, or proExM) as well as RNAs (expansion fluorescence in situ hybridization, or ExFISH), using chemicals and hardware found in a typical biology lab. Furthermore, a detailed protocol for handling and mounting expanded samples and for imaging them with confocal and light-sheet microscopes is provided. NIH (Grant 1R01NS102727) NIH (Grant 1R01EB024261) NIH (Grant 1DP1NS087724) U.S. Army Research Office (Grant W911NF1510548) U.S. Army Research Office (Grant 1RM1HG008525) IARPA (Grant D16PC00008) 2020-05-20T15:21:38Z 2020-05-20T15:21:38Z 2018-08 2019-10-02T13:00:09Z Article http://purl.org/eprint/type/JournalArticle 1934-2500 https://hdl.handle.net/1721.1/125343 Asano, Shoh M. et al. "Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues." Current Protocols in Cell Biology 80, 1 (September 2018): e56 © 2018 John Wiley & Sons, Inc. en http://dx.doi.org/10.1002/cpcb.56 Current Protocols in Cell Biology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley PMC |
| spellingShingle | Asano, Shoh M. Gao, Ruixuan Wassie, Asmamaw T. Tillberg, Paul W. Chen, Fei Boyden, Edward Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title | Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title_full | Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title_fullStr | Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title_full_unstemmed | Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title_short | Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues |
| title_sort | expansion microscopy protocols for imaging proteins and rna in cells and tissues |
| url | https://hdl.handle.net/1721.1/125343 |
| work_keys_str_mv | AT asanoshohm expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues AT gaoruixuan expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues AT wassieasmamawt expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues AT tillbergpaulw expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues AT chenfei expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues AT boydenedward expansionmicroscopyprotocolsforimagingproteinsandrnaincellsandtissues |