Review Article Page 1 of 27 Open source bioimage informatics tools for the analysis of DNA damage and associated biomarkers Jens Schneider1#, Romano Weiss1#, Madeleine Ruhe1, Tobias Jung2, Dirk Roggenbuck1,3, Ralf Stohwasser1, Peter Schierack1, Stefan Rödiger1# 1Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Senftenberg, Germany; 2Department of Molecular Toxicology (MTOX), German Institute of Human Nutrition (DIfE), 14558 Nuthetal, Germany; 3Medipan GmbH, Berlin/Dahlewitz, Germany Contributions: (I) Concept and design: J Schneider, S Rödiger; (II) Administrative support: J Schneider, S Rödiger; (III) Provision of study materials or patients: J Schneider, S Rödiger, R Weiss, R Stohwasser; (IV) Collection and assembly of data: J Schneider, S Rödiger, R Weiss; (V) Data analysis and interpretation: J Schneider, S Rödiger, R Weiss; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. #These authors contributed equally to this work. Correspondence to: Stefan Rödiger. BTU Cottbus-Senftenberg, Senftenberg, Germany. Email:
[email protected]. Abstract: DNA double-strand breaks (DSBs) are critical cellular lesions that represent a high risk for genetic instability. DSBs initiate repair mechanisms by recruiting sensor proteins and mediators. DSB biomarkers, like phosphorylated histone 2AX (γH2AX) and p53-binding protein 1 (53BP1), are detectable by immunofluorescence as either foci or distinct patterns in the vicinity of DSBs. The correlation between the number of foci and the number of DSBs makes them a useful tool for quantification of DNA damage in precision medicine, forensics and cancer research. The quantification and characterization of foci ideally uses large cell numbers to achieve statistical validity. This requires software tools that are suitable for the analysis of large complex data sets, even for users with limited experience in digital image processing.