Hyperpolarized noble gas (HNG) magnetic resonance (MR) imaging is a very promising noninvasive tool for the investigation of animal models of lung disease, particularly to follow longitudinal changes in lung function and anatomy without the accumulated radiation dose associated with x rays. The two most common noble gases for this purpose are 3He (helium 3) and 129Xe (xenon 129), the latter providing a cost-effective approach for clinical applications. Hyperpolarization is typically achieved using spin-exchange optical pumping techniques resulting in ∼10 000-fold improvement in available magnetization compared to conventional Boltzmann polarizations. This substantial increase in polarization allows high spatial resolution (≪1 mm) single-slice images of the lung to be obtained with excellent temporal resolution (≪1 s). Complete three-dimensional images of the lungs with 1 mm slice thickness can be obtained within reasonable breath-hold intervals (≪20 s). This article provides an overview of the current methods used in HNG MR imaging with an emphasis on ventilation studies in animals. Special MR hardware and software considerations are described in order to use the strong but nonrecoverable magnetization as efficiently as possible and avoid depolarization primarily by molecular oxygen. Several applications of HNG MR imaging are presented, including measurement of gross lung anatomy (e.g., airway diameters), microscopic anatomy (e.g., apparent diffusion coefficient), and a variety of functional parameters including dynamic ventilation, alveolar oxygen partial pressure, and xenon diffusing capacity.