Imaging methods that visualize the structure and function of the living body are widely used in patient care and biomedical research, but their full potential has not yet been applied to the study and treatment of the severe illnesses caused by pathogens of biodefense concern. "Conventional" imaging techniques (e.g., radiography, computed tomography, ultrasound, or magnetic resonance imaging) delineate anatomic changes in tissues, whereas "molecular" methods employ magnetic resonance, positron emission tomography, single-photon emission computed tomography, or optical (fluorescence or bioluminescence) imaging to detect biochemical reactions that accompany pathogen replication or host responses. We review the basic principles of these methods, describe the diseases caused by 6 pathogens classified as category A or B bioterror agents (anthrax, plague, tularemia, filoviral hemorrhagic fever, smallpox, and aerosolized equine encephalitis virus infection), and discuss how imaging could be used to study their pathogenesis in laboratory animals and to diagnose and monitor infection in humans.