To prospectively evaluate the correlation between a three-dimensional (3D) delayed enhancement magnetic resonance (MR) imaging sequence and a two-dimensional (2D) delayed enhancement MR imaging sequence for noninvasive assessment of myocardial viability in pigs and patients. The pig and patient studies were approved by the responsible authorities, and patients gave written informed consent. MR imaging was performed by using a rapid 3D inversion-recovery balanced steady-state free precession sequence and a 2D segmented inversion-recovery fast low-angle shot sequence as the reference standard. Fourteen pigs with reperfused (n=7) or nonreperfused (n=7) myocardial infarction and 17 patients (13 men, four women; mean age, 64.9 years+/-8.6 [standard deviation]) suspected of having myocardial infarction were included. Linear regression analysis and Bland-Altman analysis were used to compare the infarction volumes. In 10 of the 14 pigs the induction of myocardial infarction was successful. In these pigs, altogether 81 segments with myocardial infarction were demonstrated by both MR sequences, and agreement between the two sequences for classification of transmural extent of myocardial infarction was 99.7%. The infarction volume determined by using 3D MR imaging (4.64 cm3+/-2.48) in the pigs highly correlated with that of 2D MR imaging (4.65 cm3+/-2.39, r=0.989, P<.001) and that of staining by using triphenyltetrazolium chloride (4.67 cm3+/-2.44, r=0.996, P<.001). Thirteen of the 17 patients examined showed myocardial infarction in 34 myocardial segments with both sequences, and agreement between the two sequences for classification of transmural extent of myocardial infarction was 98.6%. In the patients, the infarction volume determined with both sequences highly correlated (9.71 cm3+/-7.47 for the 3D sequence vs 10.01 cm3+/-8.04 for the 2D sequence, r=0.982, P<.001). The breath-hold time necessary for the 3D MR imaging (21.0+/-2.3 seconds) was significantly shorter than that for 2D MR imaging (188.3+/-20.2 seconds, P<.001). Myocardial infarction volumes obtained with the 3D MR imaging sequence are highly correlated and in good agreement with volumes obtained with the 2D MR imaging standard approach and reduced the acquisition time by a factor of nine.