We developed a sequence by which T1- and T2*-weighted images can be acquired simultaneously and demonstrated its validity for assessing myocardial injury. The interleaved T1-T2* imaging sequence consisted of one preparatory pulse (a 90 degrees pulse) and a gradient-echo imaging sequence with a dynamically variable echo time varying between 4.2 msec for T1-weighted imaging and 15 msec for T2*-weighted imaging. The sequence was tested and validated on isolated blood-perfused pig hearts (n = 4). We found that contrast agent-induced T1 and T2* effects were clearly delineated during the first-pass and steady-state periods of a contrast agent (gadolinium diethylenetriaminopentaacetic acid). With a bolus injection of contrast agent, the maximum changes in T2* signal intensity occur significantly earlier than the changes in T1 signal. We also found that the maximum change in T1 signal intensity during the first pass of contrast agent was significantly greater in a reperfused-infarcted region than in normal regions. The suppression of T2* signal was similar in both regions. At steady state of contrast agent, T2* signal intensities gradually recovered to a significantly higher level in the reperfused-infarcted region than in normal regions. This suggests that the contrast agent diffused into the intracellular space, indicating the loss of cell membrane integrity. As a result, T1 signal intensity was also higher in the reperfused-infarcted myocardium than in normal myocardium. T1- and T2*-weighted images can be acquired simultaneously. The interleaved T1-T2* sequence is useful in assessing myocardial injury.