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

Lengert, Nicor and Mirsch, Johanna and Weimer, Ratna N. and Schumann, Eik and Haub, Peter and Drossel, Barbara and Löbrich, Markus :
AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair.
In: Scientific reports, 8 (1) p. 17282. ISSN 2045-2322
[Article] , (2018)

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.

Item Type: Article
Erschienen: 2018
Creators: Lengert, Nicor and Mirsch, Johanna and Weimer, Ratna N. and Schumann, Eik and Haub, Peter and Drossel, Barbara and Löbrich, Markus
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
Volume: 8
Number: 1
Divisions: 10 Department of Biology
10 Department of Biology > Radiation Biology and DNA Repair
05 Department of Physics
05 Department of Physics > Institute for condensed matter physics
05 Department of Physics > Institute for condensed matter physics > Statistische Physik und komplexe Systeme
Date Deposited: 26 Nov 2018 11:51
Identification Number: pmid:30470760
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