Fig. 5: In-depth imaging-based characterisation of ovarian cancer cell lines.

a Cell line characterisation workflow. Growth curves were estimated for each cell line to determine optimal seeding densities. Cells were then seeded according to estimated seeding densities into an optical, poly-L-lysine coated 96 well plate and incubated for 4 days until they reached confluence (late Log/early Stationary phase). Cells were fixed with 100% methanol and stained against indicated proteins. For each well, five independent fields were imaged using the indicated Operetta CLS™ system (40× confocal water objective). b Immunofluorescent imaging results for 73 screened cell lines ordered by the median percentage of nonmitotic cells with 2 or more centrosomes (boxplots). Boxplots show 25th, 50th and 75th centiles; whiskers indicate 75th centile plus 1.5 × inter-quartile range and 25th centile less 1.5 × inter-quartile range. . Cell lines highlighted by grey arrowheads are depicted in Fig. 6a-b. Panel 1 shows the average number of nuclei included in each of the four staining screens. Bar plots are coloured by cell line subtype. Panel 2 indicates the mitotic index for each cell line. The fraction of non-mitotic cells with two or more centrosomes is shown in panel 3 (boxplots). Each point represents an individual imaging field. The mean (population wide) CA score ( = \(\frac{\sum {Centrosomes}}{\sum {Nuclei}}\)) is shown by light blue bars. Panel 4 shows the centrosome size (estimated from max. intensity projection images), and panel 4 indicates the percentage of CEP164-positive centrosomes. Micronuclei (MN) frequencies, estimated as percentage of non-mitotic cells with at least one or more MN, are shown in panel 6. Source data are provided as a Source Data file.