Since myocardium at risk (MAR) is the major prognosticator of final infarct size and outcome in patients with acute myocardial infarction, it is highly desirable to estimate the size of the acutely ischemic myocardium, that is the MAR, in these patients. We assessed MAR size by Tc-99m-sestamibi-SPECT and computerized vectorcardiography using autoradiography as reference method. Transient myocardial ischemia was achieved in 12 pigs by coronary artery occlusion with PTCA catheters. During the procedure, computerized vectorcardiography was continuously recorded. After injection of Tc-99m-sestamibi and gadolinium-153-labelled microspheres, MAR size was estimated by SPECT and post-mortem autoradiography. Different cut-off levels (50-70%) were compared with respect to MAR-SPECT. Tc-99m-sestamibi-SPECT showed a good correlation with autoradiography (r = 0.94). Computerized vectorcardiography showed a good correlation with autoradiography as well as with Tc-99m-sestamibi-SPECT (STC-VM: r = 0.75 and 0.80, respectively, ST-VM: 0.75 and 0.87, respectively). It was found that 1) MAR assessed by Tc-99m-sestamibi-SPECT correlates closely with the autoradiographic reference; 2) a lower cut-off point of 60% of maximum uptake for MAR by Tc-99m-sestamibi-SPECT gives the closest correlation with the autoradiographic reference; and 3) ST-VM and STC-VM correlate well with MAR assessed by Tc-99m-sestamibi-SPECT and autoradiography.