Positron emission tomography (PET) is a unique tool for studying regional skeletal muscle glucose metabolism and blood flow in vivo. The application of PET in the assessment of skeletal muscle glucose metabolism depends on recent improvements in instrumentation, data analysis, and production of (18)F-fluorodeoxyglucose (FDG) and (15)O water. The data presented support the validity of the (15)O water model to measure blood flow and the FDG model for the determination of glucose uptake and glucose kinetic constants (influx, efflux, and phosphorylation) in skeletal muscle. However, quantification of absolute glucose transport and backflux rates should be applied with caution because those calculations are based on unproven assumptions such as validity of the lumped constant for these individual processes and constancy of the free and accessible intracellular glucose pool. It is evident that quantification of glucose fluxes using the triple tracer technology generates conflicting data that violate assumptions inherent in triple tracer or PET modeling. Further FDG-PET studies will have to solve those problems to provide more insight into the regulatory processes of glucose transport and phosphorylation of different insulin-resistant disease states. Promising new areas of PET research will include not only detailed study of glucose kinetics but also the measurement of muscle protein synthesis in vivo, which is of interest in a variety of conditions.