The physical considerations of using dual isotope brain SPECT imaging to monitor blood flow changes during cognitive activation studies were investigated. These factors included field uniformity, spatial resolution and crosstalk. Serial dual isotope single photon emission computed tomographic (SPECT) studies of a test tube phantom and an anthropomorphic brain phantom filled with 99Tcm and 123I were made over a 10 h period. The reconstructed counts in the 99Tcm and 123I windows were corrected for crosstalk and were plotted as a function of time. The plotted data from each window decreased over time with a half-life characteristic of each radionuclide. The relative difference between true 123I and 99Tcm region counts has to be of the order of 10% to be statistically significant at the P < 0.05 level.