A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities

Abstract Cellular responses to DNA double-strand breaks (DSBs) not only promote genomic integrity in healthy tissues, but also largely determine the efficacy of many DNA-damaging cancer treatments, including X-ray and particle therapies. A growing body of evidence suggests that activation of the mec...

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Autores principales:	Haibin Qian, Ron A. Hoebe, Michel R. Faas, Marc Jan van Goethem, Emiel R. van der Graaf, Christoph Meyer, Harry Kiewiet, Sytze Brandenburg, Przemek M. Krawczyk
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f53b3751d3154ed185a148a298d26f11
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spelling	oai:doaj.org-article:f53b3751d3154ed185a148a298d26f112021-12-02T16:14:16ZA simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities10.1038/s41598-021-92950-12045-2322https://doaj.org/article/f53b3751d3154ed185a148a298d26f112021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92950-1https://doaj.org/toc/2045-2322Abstract Cellular responses to DNA double-strand breaks (DSBs) not only promote genomic integrity in healthy tissues, but also largely determine the efficacy of many DNA-damaging cancer treatments, including X-ray and particle therapies. A growing body of evidence suggests that activation of the mechanisms that detect, signal and repair DSBs may depend on the complexity of the initiating DNA lesions. Studies focusing on this, as well as on many other radiobiological questions, require reliable methods to induce DSBs of varying complexity, and to visualize the ensuing cellular responses. Accelerated particles of different energies and masses are exceptionally well suited for this task, due to the nature of their physical interactions with the intracellular environment, but visualizing cellular responses to particle-induced damage - especially in their early stages - at particle accelerator facilities, remains challenging. Here we describe a straightforward approach for real-time imaging of early response to particle-induced DNA damage. We rely on a transportable setup with an inverted fluorescence confocal microscope, tilted at a small angle relative to the particle beam, such that cells can be irradiated and imaged without any microscope or beamline modifications. Using this setup, we image and analyze the accumulation of fluorescently-tagged MDC1, RNF168 and 53BP1—key factors involved in DSB signalling—at DNA lesions induced by 254 MeV α-particles. Our results provide a demonstration of technical feasibility and reveal asynchronous initiation of accumulation of these proteins at different individual DSBs.Haibin QianRon A. HoebeMichel R. FaasMarc Jan van GoethemEmiel R. van der GraafChristoph MeyerHarry KiewietSytze BrandenburgPrzemek M. KrawczykNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Haibin Qian Ron A. Hoebe Michel R. Faas Marc Jan van Goethem Emiel R. van der Graaf Christoph Meyer Harry Kiewiet Sytze Brandenburg Przemek M. Krawczyk A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
description	Abstract Cellular responses to DNA double-strand breaks (DSBs) not only promote genomic integrity in healthy tissues, but also largely determine the efficacy of many DNA-damaging cancer treatments, including X-ray and particle therapies. A growing body of evidence suggests that activation of the mechanisms that detect, signal and repair DSBs may depend on the complexity of the initiating DNA lesions. Studies focusing on this, as well as on many other radiobiological questions, require reliable methods to induce DSBs of varying complexity, and to visualize the ensuing cellular responses. Accelerated particles of different energies and masses are exceptionally well suited for this task, due to the nature of their physical interactions with the intracellular environment, but visualizing cellular responses to particle-induced damage - especially in their early stages - at particle accelerator facilities, remains challenging. Here we describe a straightforward approach for real-time imaging of early response to particle-induced DNA damage. We rely on a transportable setup with an inverted fluorescence confocal microscope, tilted at a small angle relative to the particle beam, such that cells can be irradiated and imaged without any microscope or beamline modifications. Using this setup, we image and analyze the accumulation of fluorescently-tagged MDC1, RNF168 and 53BP1—key factors involved in DSB signalling—at DNA lesions induced by 254 MeV α-particles. Our results provide a demonstration of technical feasibility and reveal asynchronous initiation of accumulation of these proteins at different individual DSBs.
format	article
author	Haibin Qian Ron A. Hoebe Michel R. Faas Marc Jan van Goethem Emiel R. van der Graaf Christoph Meyer Harry Kiewiet Sytze Brandenburg Przemek M. Krawczyk
author_facet	Haibin Qian Ron A. Hoebe Michel R. Faas Marc Jan van Goethem Emiel R. van der Graaf Christoph Meyer Harry Kiewiet Sytze Brandenburg Przemek M. Krawczyk
author_sort	Haibin Qian
title	A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
title_short	A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
title_full	A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
title_fullStr	A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
title_full_unstemmed	A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities
title_sort	simple microscopy setup for visualizing cellular responses to dna damage at particle accelerator facilities
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/f53b3751d3154ed185a148a298d26f11
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A simple microscopy setup for visualizing cellular responses to DNA damage at particle accelerator facilities

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