Technetium-99m sestamibi and 99mTc-tetrofosmin are at present the preferred tracers for simultaneous assessment of myocardial perfusion and function by gated single-photon emission tomography (SPET). The aim of this work was to compare sestamibi and tetrofosmin myocardial uptake 1 h after stress injection. Consecutive unselected patients were studied either with sestamibi or with tetrofosmin on a random basis, until at least 100 patients had been enrolled for each gender and tracer. Stress was obtained by dipyridamole or exercise or combined dipyridamole + exercise; in the latter cases, exercise was sustained for at least 1.5 min after tracer injection. Injected activity was similarly adjusted to body weight. For each patient, imaging began 60-75 min after injection. All SPET projections were summed; due to the acquisition technology ("roving zoom", i.e. a mobile zoom), the heart always appeared at the centre of the frame in all projections and in the sum image. Thus minimal lung background contamination could be assumed in an elliptic region of interest placed over the heart on the sum image. Three indexes were analysed: total myocardial counts (Sum), mean myocardial pixel (Mean) and maximum myocardial pixel (Max). Four patient groups were analysed: males with sestamibi or tetrofosmin (MS: n = 189 and MT: n = 157), females with sestamibi or tetrofosmin (FS: n = 101 and FT: n = 104). MS and MT groups were comparable for physical variables, maximum heart rate and stress type, as were the FS and FT groups. Sum, Mean and Max were significantly higher with sestamibi (P = 0.0001 by ANOVA). Comparing MS vs MT and FS vs FT, mean values +/- SD were as follows: for Sum (kcounts) 750+/-184 vs 652+/-166, and 707+/-202 vs 594+/-189; for Mean (counts) 4517+/-1171 vs 4107+/-898, and 4908+/-1119 vs 4144+/-1025; and for Max (counts) 6471+/-1654 vs 5794+/-1312, and 7318+/-1886 vs 6152+/-1684. The mean gain with sestamibi was +15%, +10% and +12% in males, and +19%, +18% and +19% in females. Similar differences were found within each stress type subgroup. No gender-specific effect was found for Mean, so the overall mean gain was calculated for Mean: +13% for sestamibi vs tetrofosmin. These findings are consistent with other published smaller sample series. Possible differences between tracers with regard to residual activity in syringes were ruled out by an additional experiment. In summary, we found significantly higher myocardial counts with sestamibi than with tetrofosmin, in males as well as in females.