The cellular machinery, consisting of multiple networks/pathways, is highly robust in response to genetic and environmental changes, anda living cell is capable of efficiently adjusting collective pathway activities to different cellular states and stress stimuli. The intriguing question of how such an amazing feat is achieved by different cell types as wellas in distinct cellular processes is attracting many system-level investigations and is likely to provide novel insight for deciphering complex behaviors of life. In this seminar, I will present our results from a recent study that combined quantitative single cell imaging, ensemble profiling and computational modeling to unravel systemic dynamic control over differential response to DNA damaging chemotherapeutic and chemoresistance. I will alsobriefly discuss how we extend the dynamic analysis to 3D model systems andemploy imaging-based assays to develop new drugs and drug combinations forcancer treatment.