Lengert, Nicor ; Mirsch, Johanna ; Weimer, Ratna N. ; Schumann, Eik ; Haub, Peter ; Drossel, Barbara ; Löbrich, Markus (2018)
AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair.
In: Scientific reports, 8 (1)
Artikel, Bibliographie
Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2018 |
| Autor(en): | Lengert, Nicor ; Mirsch, Johanna ; Weimer, Ratna N. ; Schumann, Eik ; Haub, Peter ; Drossel, Barbara ; Löbrich, Markus |
| Art des Eintrags: | Bibliographie |
| Titel: | AutoFoci, an automated high-throughput foci detection approach for analyzing low-dose DNA double-strand break repair. |
| Sprache: | Englisch |
| Publikationsjahr: | 23 November 2018 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Scientific reports |
| Jahrgang/Volume einer Zeitschrift: | 8 |
| (Heft-)Nummer: | 1 |
| Kurzbeschreibung (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. |
| ID-Nummer: | pmid:30470760 |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Radiation Biology and DNA Repair 05 Fachbereich Physik 05 Fachbereich Physik > Institut für Festkörperphysik (2021 umbenannt in Institut für Physik Kondensierter Materie (IPKM)) 05 Fachbereich Physik > Institut für Festkörperphysik (2021 umbenannt in Institut für Physik Kondensierter Materie (IPKM)) > Statistische Physik und komplexe Systeme |
| Hinterlegungsdatum: | 26 Nov 2018 11:51 |
| Letzte Änderung: | 03 Mai 2019 08:23 |
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