The aim of this review is to provide an update on quantification of myocardial blood flow (MBF) with positron emission tomography (PET) imaging. Technical and clinical aspects of flow quantification with PET are reviewed. The diagnostic and prognostic values of myocardial flow quantification have been established in numerous studies and in various populations. MBF quantification has also shown itself to be particularly useful in the assessment of coronary microvascular dysfunction and in evaluation of cardiac allograft vasculopathy. Overall, myocardial flow reserve (MFR) and hyperemic MBF can lead to improved risk stratification by providing information complementary to that of other markers of disease severity, such as fractional flow reserve. Flow quantification enhances MPI's ability to detect both significant epicardial disease and microvascular dysfunction. With recent technological and methodological advances, flow quantification with PET is no longer restricted to cyclotron-equipped academic centers.