Is in vivo imaging the next revolution in drug development? Current in vivo imaging modalities offer unique sensitivity, specificity and resolution for evaluating, longitudinally, drug metabolism, efficacy endpoints and off-target effects. Physiological, pathological and molecular changes can be evaluated early and rapidly. Not only does this inform the therapeutic efficacy, safety and translational viability of a drug compound, it allows the developer to make more informed decisions during the development process. Current imaging modalities offer unique sensitivity, specificity and resolution for in vivo evaluation in certain large animal models, where the close phylogenetic relation to humans is essential for safety and efficacy tests of new drugs. As such, morphological and functional imaging methods are becoming an integral part of the drug development as early and translational biomarkers in drug discovery and safety assessment. Many imaging tools are already used in preclinical research: Magnetic Resonance Imaging (MRI), Diffusion MRI, Functional MRI, Dynamic contrast-enhanced (DCE) acquisitions, Magnetic Resonance Spectroscopy, Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT). When coupled with Computed Tomography (CT), Optical imaging (fluorescence/bioluminescence) and ultrasound, we have sophisticated molecular imaging approaches that can provide much deeper insights into the molecular aspects of drug and disease targets in multiple therapeutic areas, from CNS diseases and oncology to cardiovascular, musculoskeletal and metabolic diseases. Moreover, imaging technologies have great potential to reduce development time, animal use and improve the efficiency of the drug development process.