High-content analysis (HCA) provides the ability to study many cellular features simultaneously and has the power to revolutionize drug discovery, especially as it is applied to more physiologically relevant models. Complex biological pathways may be explored, drug efficacy predicted, and ultimately, attrition rate reduced; however, challenges remain as these complex assays push the field forward. Cambridge Healthtech Institute’s High-Content Analysis conference will cover applications of HCA in preclinical drug discovery, challenges of the high-content screening environment, and HCS in immuno-oncology.