Positron emission tomography (PET) is a nuclear imaging technique that can dynamically image trace amounts of positron-labeled radiopharmaceuticals in vivo. Tracer concentrations can be determined quantitatively, and by application of appropriate tracer kinetic models, the rates of a wide range of different biological processes can be measured noninvasively in humans. PET has been used as a research tool for more than 25 years and has also found clinical applications, particularly in oncology, neurological disorders, and cardiovascular disease. Recently, there has been tremendous interest in applying PET technology to in vivo small-animal imaging. Significant improvements in the imaging technology now permit a wide range of PET studies in mice and rats, using compact, relatively low-cost, dedicated small-animal PET scanners. This article reviews the fundamental basis of PET imaging and discusses the development of small-animal PET scanners and their possible application in preclinical drug development.