Many different types of radiation have been exploited to provide images of the structure and function of tissues inside a living subject. Each imaging modality is characterized by differing resolutions on the spatial and temporal scales, and by a different sensitivity for measuring properties related to morphology or function. Combinations of imaging modalities that integrate the strengths of two modalities, and at the same time eliminate one or more weaknesses of an individual modality, thus offer the prospect of improved diagnostics, therapeutic monitoring, and preclinical research using imaging approaches. This review discusses the advantages and challenges in developing multimodality imaging systems for in vivo use, highlights some successful combinations that are now routinely used in the clinic and in research, and discusses recent advances in multimodality instrumentation that may offer new opportunities for imaging.