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AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair

Lengert, Nicor ; Mirsch, Johanna ; Weimer, Ratna N. ; Schumann, Eik ; Haub, Peter ; Drossel, Barbara ; Löbrich, Markus (2021):
AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair. (Publisher's Version)
In: Scientific Reports, 8 (1), Springer Nature, e-ISSN 2045-2322,
DOI: 10.26083/tuprints-00019035,
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Item Type: Article
Origin: Secondary publication service
Status: Publisher's Version
Title: AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair
Language: English
Abstract:

Double-strand breaks (DSBs) are the most lethal DNA damages induced by ionising radiation (IR) and their efficient repair is crucial to limit genomic instability. The cellular DSB response after low IR doses is of particular interest but its examination requires the analysis of high cell numbers. Here, we present an automated DSB quantification method based on the analysis of γH2AX and 53BP1 foci as markers for DSBs. We establish a combination of object properties, combined in the object evaluation parameter (OEP), which correlates with manual object classification. Strikingly, OEP histograms show a bi-modal distribution with two maxima and a minimum in between, which correlates with the manually determined transition between background signals and foci. We used algorithms to detect the minimum, thus separating foci from background signals and automatically assessing DSB levels. To demonstrate the validity of this method, we analyzed over 600.000 cells to verify results of previous studies showing that DSBs induced by low doses are less efficiently repaired compared with DSBs induced by higher doses. Thus, the automated foci counting method, called AutoFoci, provides a valuable tool for high-throughput image analysis of thousands of cells which will prove useful for many biological screening approaches.

Journal or Publication Title: Scientific Reports
Journal volume: 8
Number: 1
Publisher: Springer Nature
Collation: 11 Seiten
Classification DDC: 500 Naturwissenschaften und Mathematik > 530 Physik
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Divisions: 10 Department of Biology > Radiation Biology and DNA Repair
05 Department of Physics > Institute for Condensed Matter Physics
Date Deposited: 17 Aug 2021 12:08
Last Modified: 17 Aug 2021 12:08
DOI: 10.26083/tuprints-00019035
Corresponding Links:
URN: urn:nbn:de:tuda-tuprints-190353
URI: https://tuprints.ulb.tu-darmstadt.de/id/eprint/19035
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