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Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells
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 s...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7286065/ https://www.ncbi.nlm.nih.gov/pubmed/32497404 http://dx.doi.org/10.1002/cpns.96 |
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author | Zhang, Chi Kang, Jeong Seuk Asano, Shoh M. Gao, Ruixuan Boyden, Edward S. |
author_facet | Zhang, Chi Kang, Jeong Seuk Asano, Shoh M. Gao, Ruixuan Boyden, Edward S. |
author_sort | Zhang, Chi |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7286065 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-72860652021-06-01 Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells Zhang, Chi Kang, Jeong Seuk Asano, Shoh M. Gao, Ruixuan Boyden, Edward S. Curr Protoc Neurosci Protocol 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. John Wiley and Sons Inc. 2020-06-04 2020-06 /pmc/articles/PMC7286065/ /pubmed/32497404 http://dx.doi.org/10.1002/cpns.96 Text en © 2020 The Authors. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Protocol 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 |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7286065/ https://www.ncbi.nlm.nih.gov/pubmed/32497404 http://dx.doi.org/10.1002/cpns.96 |
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