Figure 3.

Multi-content 3D readout to test drug combinations. OvCa cells were seeded in ultra-low attachment (ULA) plates and spheroids formed four days before treatment. Daily imaging using NYONE^® Scientific (SYNENTEC) recorded the formation process and spheroid morphology and growth over time. On day four, spheroids were treated with cisplatin (asterisks) in absence or presence of metalloprotease inhibitors (blue circles), as described. To allow multiplex readout of three consecutive assays in one 96-well plate, the real-time fluorescence-based cytotoxicity assay CellTox^TM Green was first applied on day five and imaged twice: day five = 24 h after treatment, and day six = 48 h after treatment. Next, cell viability and caspase activation were measured using two different luminescent assays to allow for multiplexing using different filters.