Segmentation and quantitation pipeline for image analysis from Targeting RNA-Polymerase I in Both Chemosensitive and Chemoresistant Populations in Epithelial Ovarian Cancer

Explanation and example analysis pipeline used to quantitate changes in cell size and other parameters. Supplemental Figure 1: (A) Sample images of immunofluorescence of F-actin (red) and DAPI (blue) before and after IC50treatment with CX-5461. Images were taken at random at 200x magnification. Thresholding was performed to identify DAPI stained nuclei using the Mixture of Gaussian (MoG) technique to bin pixels into foreground and background classes. Cytoplasm, marked by F-actin, was segmented using the watershed technique after Sobel transformation followed by Otsu thresholding. All image segmentation and quantitation was performed by CellProfiler. (B) Summary quantitation in representative images of changes to segmented cells' parameters before and after treatment.