The calculation of ejection fraction using gated single photon emission computed tomography (SPECT) has been widely validated against a range of other techniques. There have been fewer studies validating left ventricular volumes. We compared quantitative gated SPECT (QGS) with magnetic resonance imaging (MRI) measurements of left ventricular ejection fraction and end diastolic volume in 50 patients with a large range of ventricular dimensions. MRI data were obtained using a turbo gradient echo pulse sequence (TGE) in 17 patients and a steady state free precession pulse sequence (SSFP) in 33 patients. There was good correlation between ejection fraction and end diastolic volume measurements from SPECT and MRI (r=0.82, r=0.90, respectively) but the mean SPECT values were significantly lower (ejection fraction, 6.6+/-6.4% points; end diastolic volume, 18.4+/-25.4 ml) than those obtained from MRI. Bland-Altman analysis showed some large differences in individual patients but no trends in the data either in ejection fraction over a range from 15% to 70% or in end diastolic volume, range 75-400 ml. SSFP gave a larger difference for end diastolic volume measurement compared to SPECT than did TGE, although this difference did not reach significance. Both SSFP and TGE gave similar values for the difference between MRI and SPECT for the measurement of ejection fraction. We suggest that the difference in EF may be a result of 8 frames being used for gating in QGS but 12-18 for MR. Differences in volumes may be related to the different spatial resolution and the exclusion or inclusion of trabeculation and papillary muscles between SPECT and MRI. Differences between SSFP and TGE may be caused by differing delineation of the endocardial border, dependent on the particular acquisition sequence. In conclusion, QGS values correlated well with MRI, but a correction factor may be needed if direct comparison is made.