MRI provides non-invasive and non-ionizing methods for the accurate anatomic depiction of the cardiovascular system. Based on the inherent flow sensitivity, MRI can be used to investigate hemodynamic features in patients with anatomical data within a single measurement. In particular, time-resolved and three-dimensional (3D) characterization of blood flow using 4D flow MRI has achieved considerable progress in recent years. The present article reviews the principle and procedures of 4D Flow MRI. Various fluid dynamic biomarkers for possible clinical usage are also described, including wall shear stress, turbulent kinetic energy, and relative pressure. Finally, this article provides an overview of the clinical applications of 4D Flow MRI in various cardiovascular regions.