Positron emission tomography (PET) is an imaging technology that measures 3D spatial distribution and kinetics of radio-tagged biomolecules in a living subject quantitatively and nondestructively. Commonly used positron-emitting radionuclides include11C,13N, and15O, which are essential elements for plant growth. Combining radiotracer techniques with PET, this in vivo molecular imaging capability offers plant biologists a powerful tool for molecular phenotyping research. While PET is widely used clinically for cancer diagnosis and pre-clinically for drug development, it is an unfamiliar imaging tool for plant biologists. This chapter introduces the basic principles of PET, factors that affect the quantitative accuracy of PET when imaging plants, and techniques for administering radiotracers to plants for a variety of molecular plant imaging applications.