We hypothesized that tissue harmonic imaging (THI) in comparison with fundamental imaging (FI) would improve endocardial border detection, and therefore in combination with 3-dimensional echocardiography (3D echo), it would be a precise method for left ventricular (LV) volume measurement. Ten healthy subjects and 18 consecutive patients with dilated hearts underwent estimation of LV volumes by magnetic resonance imaging (MRI) and transthoracic 3D echo with THI and FI. In patients, the agreement between MRI and 3D echo was closer with THI in comparison with FI for assessment of LV volumes. Thus the mean +/- 2 SD of differences between MRI and 3D echo with THI versus FI, respectively, was -6.4 +/- 40.0 mL versus -17.4 +/- 57.6 mL (P <.01) for the end-diastolic volume (EDV), and 0.0 +/- 26.6 mL versus -8.1 +/- 35.6 mL (P <.01) for the end-systolic volume (ESV). In patients, THI in comparison with FI approximately halved observer variation on EDV and ESV. In healthy subjects, only ESV showed significantly reduced observer variation by THI. In conclusion, because THI demonstrated a clinically relevant reduction in observer variation and a closer agreement to the MRI technique in patients with dilated hearts, it should replace FI in LV volume measurements.