Fig. 5.

Implementation of DNA damage prediction model generated from 2D organoids show agreement in 3D organoids. 3D organoids from a treatment naïve patient were cultured in 80 % Matrigel and were continuously treated over 5 days in 10 μm of etoposide, bortezomib, olaparib, or cisplatin. a) Representative 3PF, 2PF, THG, and optical redox ratio SLAM images from before treatment, 2 days and 5 days post-treatment. (scalebar: 50 μm) b) Representative immunofluorescence imaging of DAPI and γH2AX signal in treated organoids (scalebar: 50 μm). c) Normalized ground truth data after 2 and 5 days of treatment (3 biological replicates with 16–25 technical replicates per biological replicate). d) Predicted DNA damage after days 2 and 5 of treatment based on SLAM images show agreement with ground truth data (4 biological replicates with 9 technical replicates per biological replicate). e) AUC-RR quantification from SLAM imaging used to inform predictions in figure d. c,d) Statistical testing was performed using ANOVA followed by Tukey's post-hoc test or Kruskal-Wallis followed by Dunn's post-hoc test across all groups within each timepoint. Dotted boxes were used to denote shared significance from media control (*p < 0.05, **p < 0.01, ***p < 0.001).