A computational approach to quantifying miscounting of radiation-induced double-strand break immunofluorescent foci
Name:
35835982.pdf
Size:
3.094Mb
Format:
PDF
Description:
Identified with Open Access button
Authors
Ingram, SamuelWarmenhoven, John W
Henthorn, Nicholas
Chadwick, Amy
Santina, Elham
McMahon, S. J.
Schuemann, J.
Kirkby, Norman
Mackay, Ranald I
Kirkby, Karen J
Merchant, Michael J
Affiliation
Division of Cancer Sciences, Faculty of Biology, Medicine and Health, The University of Manchester, Oxford Rd, Manchester, M13 9PL, UKIssue Date
2022
Metadata
Show full item recordAbstract
Immunofluorescent tagging of DNA double-strand break (DSB) markers, such as γ-H2AX and other DSB repair proteins, are powerful tools in understanding biological consequences following irradiation. However, whilst the technique is widespread, there are many uncertainties related to its ability to resolve and reliably deduce the number of foci when counting using microscopy. We present a new tool for simulating radiation-induced foci in order to evaluate microscope performance within in silico immunofluorescent images. Simulations of the DSB distributions were generated using Monte Carlo track-structure simulation. For each DSB distribution, a corresponding DNA repair process was modelled and the un-repaired DSBs were recorded at several time points. Corresponding microscopy images for both a DSB and (γ-H2AX) fluorescent marker were generated and compared for different microscopes, radiation types and doses. Statistically significant differences in miscounting were found across most of the tested scenarios. These inconsistencies were propagated through to repair kinetics where there was a perceived change between radiation-types. These changes did not reflect the underlying repair rate and were caused by inconsistencies in foci counting. We conclude that these underlying uncertainties must be considered when analysing images of DNA damage markers to ensure differences observed are real and are not caused by non-systematic miscounting.Citation
Ingram SP, Warmenhoven JW, Henthorn NT, Chadiwck AL, Santina EE, McMahon SJ, et al. A computational approach to quantifying miscounting of radiation-induced double-strand break immunofluorescent foci. Communications biology. 2022 Jul 14;5(1):700. PubMed PMID: 35835982. Pubmed Central PMCID: PMC9283546. Epub 2022/07/15. eng.Journal
Communications BiologyDOI
10.1038/s42003-022-03585-5PubMed ID
35835982Additional Links
https://dx.doi.org/10.1038/s42003-022-03585-5Type
ArticleLanguage
enae974a485f413a2113503eed53cd6c53
10.1038/s42003-022-03585-5
Scopus Count
Collections
Related articles
- Computational modelling of γ-H2AX foci formation in human cells induced by alpha particle exposure.
- Authors: Abu Shqair A, Lee US, Kim EH
- Issue date: 2022 Aug 23
- Kinetics and dose-response of residual 53BP1/gamma-H2AX foci: co-localization, relationship with DSB repair and clonogenic survival.
- Authors: Marková E, Schultz N, Belyaev IY
- Issue date: 2007 May
- Predicting Radiosensitivity with Gamma-H2AX Foci Assay after Single High-Dose-Rate and Pulsed Dose-Rate Ionizing Irradiation.
- Authors: van Oorschot B, Hovingh S, Dekker A, Stalpers LJ, Franken NA
- Issue date: 2016 Feb
- Impact of dimethyl sulfoxide on irradiation-related DNA double-strand-break induction, -repair and cell survival.
- Authors: Zwicker F, Hauswald H, Debus J, Huber PE, Weber KJ
- Issue date: 2019 Aug
- The influence of heterochromatin on DNA double strand break repair: Getting the strong, silent type to relax.
- Authors: Goodarzi AA, Jeggo P, Lobrich M
- Issue date: 2010 Dec 10