To streamline the identification of potentially active cancer therapeutics, here we describe a highly adaptable semi-automated approach to screen compounds simultaneously across a panel of cell lines. These protocols leverage automation to enhance robustness, reproducibility, and throughput while integrating the IncuCyte ZOOM live-cell imaging platform and the CellTiter-Glo endpoint viability assay to assess drug efficacy. The integration of both evaluative methods strategically bypasses the shortcomings of each approach individually allowing for more thorough and detailed analysis within a single drug screen. The expected output from protocol utilization includes traditional dose-response curves, IC50, area-under the curve (AUC), GR50, area-over the curve (AOC) as well as live-cell imaging to identify cell specific morphological changes, cytostatic, or cytotoxic effects of 72-hour drug treatments. This adaptable protocol can be employed across cancer model systems and represents a reproducible procedure to optimize 96-well plate cell growth conditions compatible with the integrated drug screen and simultaneously assess drug efficacy across multiple cell lines in future cancer research studies.

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