Absolute myocardial blood flow (MBF) and myocardial flow reserve (MFR) provide incremental diagnostic and prognostic information over relative perfusion alone. Recent development of dedicated cardiac SPECT cameras with better sensitivity and temporal resolution make dynamic SPECT imaging more practical. In this study, we evaluate the measurement of MBF using a multipinhole dedicated cardiac SPECT camera in a pig model of rest and transient occlusion at stress using 3 common tracers: 201 Tl, 99m Tc-tetrofosmin, and 99m Tc-sestamibi. Methods: Animals (n 5 19) were injected at rest/stress with 99m Tc radiotracers (370/1,100 MBq) or 201 Tl (37/110 MBq) with a 1-h delay between rest and dipyridamole stress. With each tracer, microspheres were injected simultaneously as the gold standard measurement for MBF. Dynamic images were obtained for 11 min starting with each injection. Residual resting activity was subtracted from stress data and images reconstructed with CT-based attenuation correction and energy window-based scatter correction. Dynamic images were processed with kinetic analysis software using a 1-tissue-compartment model to obtain the uptake rate constant K 1 as a function of microsphere MBF. Results: Measured extraction fractions agree with those obtained previously using ex vivo techniques. Converting K 1 back to MBF using the measured extraction fractions produced accurate values and good correlations with microsphere MBF: r 5 0.75-0.90 (P , 0.01 for all). The correlation in the MFR was between r 5 0.57 and 0.94 (P , 0.01). Conclusion: Noninvasive measurement of absolute MBF with stationary dedicated cardiac SPECT is feasible using common perfusion tracers. St udies using PET have demonstrated that absolute myocardial blood flow (MBF) and myocardial flow reserve (MFR 5 stress/ rest MBF) provide incremental diagnostic and prognostic information over relative perfusion alone (1-4). Imaging of myocardial perfusion is much more commonly performed with SPECT than with PET, but MBF measurements are not typically acquired.Measuring MBF is difficult with standard SPECT cameras because of the need for attenuation and scatter correction and the need to rotate around the patient for 3-dimensional imaging. Recent studies have shown that it is possible to obtain an index of the MFR without a direct measure of MBF using a combination of dynamic planar followed by static SPECT acquisitions (5) and that this can provide some prognostic information (6). Other studies have demonstrated that rapid camera rotation can provide dynamic tomographic data and hence a measure of MFR (7) and the arterial input function (8), suggesting that accurate measures of MBF could be possible (9). The practicality of measuring MBF has increased greatly, however, with the advent of dedicated cardiac cameras.Dedicated cardiac cameras such as the DSPECT system (Spectrum Dynamics Medical Inc.) or the Discovery NM 530c/ 570c cameras (GE Healthcare) have greatly improved sensitivity and do not rotate around the patient (10). These features allow d...