Evaluating Cell Viability and Growth under Oxaliplatin Exposure

End point growth rate experiments of HCT116, HT29, and Caco2 were carried out in 96-well CellCarrier plates (PerkinElmer No. 6005558, Waltham, MA, USA) at an initial cell seeding of 1,500, 4,000, and 2,000 cells per well, respectively. One day after seeding, cells were treated with the indicated dilutions of oxaliplatin (Selleck Chemicals No. S1224, Houston, TX, USA). At the stated time points, images were acquired on an Operetta HCS System (PerkinElmer No. HH12000000) equipped with environmental controls (37°C, 5% CO₂). Thirty minutes prior to imaging, cells were stained with 5 μg/mL of Hoechst 33342 (Invitrogen No. H21492, Carlsbad, CA, USA) and 5 μg/mL of propidium iodine (Invitrogen No. P1304MP, Carlsbad, CA, USA) to determine live or dead cells, respectively. For live cell experiments, cells were seeded on 0.2 or 2 kPa softwell (Matrigen) or CellCarrier plates in the presence or absence of liver ECM disc. Images were taken on the Operetta HCS in confocal mode using the z-stack function. For all experiments, image analysis was performed using the Harmony 3.5.2 software (PerkinElmer No. HH17000001, Waltham, MA, USA). Cells were identified and segmented at the nuclear level to determine live and dead cell counts over time as described previously [32 (link)].