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Analysis of DNA damage is key for cancer research. High-content screening allows testing the effect of different conditions on genotoxicity in an efficient manner.
After acquiring the data, as a first step, the DAPI nuclei are segmented. Intensity measurements of another nuclear signal (red signal) determines the cell cycle stage. It is then possible to classify these nuclei based on intracellular DNA-damage foci (EdU, green signal) using a parent-child operation in the analysis pipeline.
Figure 1: Nuclei (blue) with cell cycle marker (red) DNA damage foci (green).
Figure 2: Segmented nuclei (cyan highlights) and segmented foci (magenta highlights) within each nucleus.
The solution allows for fast and flexible stratification of nuclei according to a diverse array of parameters for an in-depth mechanistic analysis of genotoxicity. Once set up, such an analysis can easily be scaled up to hundreds of samples.
Figure 3: Scatter plot showing DNA damage marker intensity (EdU) over DAPI intensity for each nucleus. Expected values for different cell cycle phases are highlighted in red.