Given a pharmaceutical landscape in which fewer drugs are succeeding in reaching the market, pharmaceutical and biotechnological companies are seeking alternative screening methodologies that will be compatible with the large scale of current combinatorial chemical libraries. In this context, HCS has received considerable attention. Imaging technologies are playing an increasing role in the drug discovery and development process, and this role is projected to increase further in the future. Currently, these technologies are rarely applied in primary screening campaigns but, rather, are used in the processes that precede and follow primary screening. Imaging technologies are employed for target identification and validation, secondary screening, ADMET studies, and pharmacokinetic studies. Various labeling technologies are deployed for such imaging, including fluorescence, luminescence, PET and computer tomography (CT). This feature review discusses high-content analysis (HCA), including the HCS technology and methodology involved, and the future potential of HCA in the drug discovery process.