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  1. DNA double-strand breaks (DSBs) occur frequently in eukaryotic cells, and the homologous recombination pathway (HR) is one of the major pathways required to repair these breaks. However, tumor cells that are able to repair DSBs are unlikely to die due to damage incurred by DNA damaging chemotherapies, such as platinum compounds. While platinum-based therapies have been effective in treating various cancers, they also carry harsh side effects, and thus ideally platinum should be used when the probability of treatment resistance is low. HR scores provide a measure for patients’ tumor’s HR capacity and have been shown to predict their chemotherapy response and long-term survival. Calculating this score manually from immunofluorescence microscopy images for each patient is error-prone and time-consuming. Herein, we propose an image processing pipeline that takes as input imaging data from three emission channels (representing nuclei, S-phase cells, and HR-mediated repair in a tumor slice) from an epifluorescence microscope and computes the HR score. Our open-source methodology forms a rationale to develop similar approaches in predicting chemotherapeutic responses and facilitating to make treatment decisions. 
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