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Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells
Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former la...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8647181/ https://www.ncbi.nlm.nih.gov/pubmed/34866605 http://dx.doi.org/10.1107/S2059798321010329 |
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author | Koronfel, Mohamed Kounatidis, Ilias Mwangangi, Dennis M. Vyas, Nina Okolo, Chidinma Jadhav, Archana Fish, Tom Chotchuang, Phatcharin Schulte, Albert Robinson, Robert C. Harkiolaki, Maria |
author_facet | Koronfel, Mohamed Kounatidis, Ilias Mwangangi, Dennis M. Vyas, Nina Okolo, Chidinma Jadhav, Archana Fish, Tom Chotchuang, Phatcharin Schulte, Albert Robinson, Robert C. Harkiolaki, Maria |
author_sort | Koronfel, Mohamed |
collection | PubMed |
description | Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former lacks the capacity to capture the overall local ultrastructure, while the latter requires rigorous sample preparation that can lead to potential artefacts, and only delivers relatively small volumes of imaging data at the thinnest areas of a cell. In this work, a correlative approach utilizing in situ super-resolution fluorescence imaging and cryo X-ray tomography was used to image bundles of actin filaments deep inside cells under near-native conditions. In this case, fluorescence 3D imaging localized the actin bundles within the intracellular space, while X-ray tomograms of the same areas provided detailed views of the local ultrastructure. Using this new approach, actin trails connecting vesicles in the perinuclear area and hotspots of actin presence within and around multivesicular bodies were observed. The characteristic prevalence of filamentous actin in cytoplasmic extensions was also documented. |
format | Online Article Text |
id | pubmed-8647181 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-86471812021-12-16 Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells Koronfel, Mohamed Kounatidis, Ilias Mwangangi, Dennis M. Vyas, Nina Okolo, Chidinma Jadhav, Archana Fish, Tom Chotchuang, Phatcharin Schulte, Albert Robinson, Robert C. Harkiolaki, Maria Acta Crystallogr D Struct Biol Ccp4 Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former lacks the capacity to capture the overall local ultrastructure, while the latter requires rigorous sample preparation that can lead to potential artefacts, and only delivers relatively small volumes of imaging data at the thinnest areas of a cell. In this work, a correlative approach utilizing in situ super-resolution fluorescence imaging and cryo X-ray tomography was used to image bundles of actin filaments deep inside cells under near-native conditions. In this case, fluorescence 3D imaging localized the actin bundles within the intracellular space, while X-ray tomograms of the same areas provided detailed views of the local ultrastructure. Using this new approach, actin trails connecting vesicles in the perinuclear area and hotspots of actin presence within and around multivesicular bodies were observed. The characteristic prevalence of filamentous actin in cytoplasmic extensions was also documented. International Union of Crystallography 2021-11-29 /pmc/articles/PMC8647181/ /pubmed/34866605 http://dx.doi.org/10.1107/S2059798321010329 Text en © Mohamed Koronfel et al. 2021 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Ccp4 Koronfel, Mohamed Kounatidis, Ilias Mwangangi, Dennis M. Vyas, Nina Okolo, Chidinma Jadhav, Archana Fish, Tom Chotchuang, Phatcharin Schulte, Albert Robinson, Robert C. Harkiolaki, Maria Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title | Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title_full | Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title_fullStr | Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title_full_unstemmed | Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title_short | Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells |
title_sort | correlative cryo-imaging of the cellular universe with soft x-rays and laser light used to track f-actin structures in mammalian cells |
topic | Ccp4 |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8647181/ https://www.ncbi.nlm.nih.gov/pubmed/34866605 http://dx.doi.org/10.1107/S2059798321010329 |
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