An emission computed tomography system (SPECT), which uses a single large-field-of-view gamma camera, was evaluated for its ability to measure the relative distribution of myocardial blood flow and to assess the effect of attenuation, scatter, and cardiac motion on the tomographic images. Normalized regional myocardial counts from the SPECT images of the living dogs correlated closely with those from the anatomic slices and the samples counted at necropsy except for an over-estimate of tracer in the perfusion defect (SPECT) 57.7 compared to tissue count 32.1; p less than 0.05. The differences were less for the other imaging conditions. Heart and thorax motion, attenuation, and scatter contributed less than 25% to the over-estimate of defect counts. We conclude that the SPECT system accurately reflects regional distribution of myocardial blood flow except for overestimation of flow in regions of perfusion defects. Small perfusion defects might therefore be missed, but no artifactual defects are created.