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Imaging of Virus-Infected Cells with Soft X-ray Tomography
Viruses are obligate parasites that depend on a host cell for replication and survival. Consequently, to fully understand the viral processes involved in infection and replication, it is fundamental to study them in the cellular context. Often, viral infections induce significant changes in the subc...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8618346/ https://www.ncbi.nlm.nih.gov/pubmed/34834916 http://dx.doi.org/10.3390/v13112109 |
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author | Garriga, Damià Chichón, Francisco Javier Calisto, Bárbara M. Ferrero, Diego S. Gastaminza, Pablo Pereiro, Eva Pérez-Berna, Ana Joaquina |
author_facet | Garriga, Damià Chichón, Francisco Javier Calisto, Bárbara M. Ferrero, Diego S. Gastaminza, Pablo Pereiro, Eva Pérez-Berna, Ana Joaquina |
author_sort | Garriga, Damià |
collection | PubMed |
description | Viruses are obligate parasites that depend on a host cell for replication and survival. Consequently, to fully understand the viral processes involved in infection and replication, it is fundamental to study them in the cellular context. Often, viral infections induce significant changes in the subcellular organization of the host cell due to the formation of viral factories, alteration of cell cytoskeleton and/or budding of newly formed particles. Accurate 3D mapping of organelle reorganization in infected cells can thus provide valuable information for both basic virus research and antiviral drug development. Among the available techniques for 3D cell imaging, cryo–soft X-ray tomography stands out for its large depth of view (allowing for 10 µm thick biological samples to be imaged without further thinning), its resolution (about 50 nm for tomographies, sufficient to detect viral particles), the minimal requirements for sample manipulation (can be used on frozen, unfixed and unstained whole cells) and the potential to be combined with other techniques (i.e., correlative fluorescence microscopy). In this review we describe the fundamentals of cryo–soft X-ray tomography, its sample requirements, its advantages and its limitations. To highlight the potential of this technique, examples of virus research performed at BL09-MISTRAL beamline in ALBA synchrotron are also presented. |
format | Online Article Text |
id | pubmed-8618346 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-86183462021-11-27 Imaging of Virus-Infected Cells with Soft X-ray Tomography Garriga, Damià Chichón, Francisco Javier Calisto, Bárbara M. Ferrero, Diego S. Gastaminza, Pablo Pereiro, Eva Pérez-Berna, Ana Joaquina Viruses Review Viruses are obligate parasites that depend on a host cell for replication and survival. Consequently, to fully understand the viral processes involved in infection and replication, it is fundamental to study them in the cellular context. Often, viral infections induce significant changes in the subcellular organization of the host cell due to the formation of viral factories, alteration of cell cytoskeleton and/or budding of newly formed particles. Accurate 3D mapping of organelle reorganization in infected cells can thus provide valuable information for both basic virus research and antiviral drug development. Among the available techniques for 3D cell imaging, cryo–soft X-ray tomography stands out for its large depth of view (allowing for 10 µm thick biological samples to be imaged without further thinning), its resolution (about 50 nm for tomographies, sufficient to detect viral particles), the minimal requirements for sample manipulation (can be used on frozen, unfixed and unstained whole cells) and the potential to be combined with other techniques (i.e., correlative fluorescence microscopy). In this review we describe the fundamentals of cryo–soft X-ray tomography, its sample requirements, its advantages and its limitations. To highlight the potential of this technique, examples of virus research performed at BL09-MISTRAL beamline in ALBA synchrotron are also presented. MDPI 2021-10-20 /pmc/articles/PMC8618346/ /pubmed/34834916 http://dx.doi.org/10.3390/v13112109 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Review Garriga, Damià Chichón, Francisco Javier Calisto, Bárbara M. Ferrero, Diego S. Gastaminza, Pablo Pereiro, Eva Pérez-Berna, Ana Joaquina Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title | Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title_full | Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title_fullStr | Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title_full_unstemmed | Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title_short | Imaging of Virus-Infected Cells with Soft X-ray Tomography |
title_sort | imaging of virus-infected cells with soft x-ray tomography |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8618346/ https://www.ncbi.nlm.nih.gov/pubmed/34834916 http://dx.doi.org/10.3390/v13112109 |
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