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Magnetic resonance microscopy (MRM) has become an important tool for small animal cardiac imaging. In relation to competing technologies (microCT and ultrasound), MR is limited by spatial resolution, temporal resolution, and acquisition time. All three of these limitations have been addressed by developing a four-dimensional (4D) (3D plus time) radial acquisition (RA) sequence. The signal-to-noise ratio (SNR) has been optimized by minimizing the echo time (TE) (300 us). The temporal resolution and throughput have been improved by center-out trajectories resulting in repetition time (TR) <2.5 ms. The contrast has been enhanced through the use of a liposomal blood pool agent that reduces theT1 of the blood to <400 ms. We have developed protocols for three specific applications: 1) high-throughput with spatial resolution of 87 × 87 × 352 um3 (voxel volume = 2.7 nL) and acquisition time of 16 min; 2) high-temporal resolution with spatial resolution of 87 × 87 × 352 um3 (voxel volume = 2.7 nL) and temporal resolution at 4.8 ms and acquisition time of 32 minutes; and 3) high-resolution isotropic imaging at 87 × 87 × 87 um3 (voxel volume = 0.68 nL) and acquisition time of 31 min. The 4D image arrays allow direct measure of cardiac functional parameters dependent on chamber volumes, e.g., ejection fraction (EF), end diastolic volume (EDV), and end systolic volume (ESV). Magn Reson Med 60:111–118, 2008. © 2008 Wiley-Liss, Inc.