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Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution

Expansion microscopy is a sample preparation technique that enables the optical imaging of biological specimens at super-resolution owing to their physical magnification, which is achieved through water-absorbing polymers. The technique uses readily available chemicals and does not require sophistic...

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Detalles Bibliográficos
Autores principales: Mascheroni, Luca, Scherer, Katharina M., Manton, James D., Ward, Edward, Dibben, Oliver, Kaminski, Clemens F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Optical Society of America 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7510880/
https://www.ncbi.nlm.nih.gov/pubmed/33014598
http://dx.doi.org/10.1364/BOE.399404
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author Mascheroni, Luca
Scherer, Katharina M.
Manton, James D.
Ward, Edward
Dibben, Oliver
Kaminski, Clemens F.
author_facet Mascheroni, Luca
Scherer, Katharina M.
Manton, James D.
Ward, Edward
Dibben, Oliver
Kaminski, Clemens F.
author_sort Mascheroni, Luca
collection PubMed
description Expansion microscopy is a sample preparation technique that enables the optical imaging of biological specimens at super-resolution owing to their physical magnification, which is achieved through water-absorbing polymers. The technique uses readily available chemicals and does not require sophisticated equipment, thus offering super-resolution to laboratories that are not microscopy-specialised. Here we present a protocol combining sample expansion with light sheet microscopy to generate high-contrast, high-resolution 3D reconstructions of whole virus-infected cells. The results are superior to those achievable with comparable imaging modalities and reveal details of the infection cycle that are not discernible before expansion. An image resolution of approximately 95 nm could be achieved in samples labelled in 3 colours. We resolve that the viral nucleoprotein is accumulated at the membrane of vesicular structures within the cell cytoplasm and how these vesicles are positioned relative to cellular structures. We provide detailed guidance and a video protocol for the optimal application of the method and demonstrate its potential to study virus-host cell interactions.
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spelling pubmed-75108802020-10-01 Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution Mascheroni, Luca Scherer, Katharina M. Manton, James D. Ward, Edward Dibben, Oliver Kaminski, Clemens F. Biomed Opt Express Article Expansion microscopy is a sample preparation technique that enables the optical imaging of biological specimens at super-resolution owing to their physical magnification, which is achieved through water-absorbing polymers. The technique uses readily available chemicals and does not require sophisticated equipment, thus offering super-resolution to laboratories that are not microscopy-specialised. Here we present a protocol combining sample expansion with light sheet microscopy to generate high-contrast, high-resolution 3D reconstructions of whole virus-infected cells. The results are superior to those achievable with comparable imaging modalities and reveal details of the infection cycle that are not discernible before expansion. An image resolution of approximately 95 nm could be achieved in samples labelled in 3 colours. We resolve that the viral nucleoprotein is accumulated at the membrane of vesicular structures within the cell cytoplasm and how these vesicles are positioned relative to cellular structures. We provide detailed guidance and a video protocol for the optimal application of the method and demonstrate its potential to study virus-host cell interactions. Optical Society of America 2020-08-14 /pmc/articles/PMC7510880/ /pubmed/33014598 http://dx.doi.org/10.1364/BOE.399404 Text en Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License (http://creativecommons.org/licenses/by/4.0/) . Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.
spellingShingle Article
Mascheroni, Luca
Scherer, Katharina M.
Manton, James D.
Ward, Edward
Dibben, Oliver
Kaminski, Clemens F.
Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title_full Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title_fullStr Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title_full_unstemmed Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title_short Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
title_sort combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7510880/
https://www.ncbi.nlm.nih.gov/pubmed/33014598
http://dx.doi.org/10.1364/BOE.399404
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