Definition and validation of the ranges of normal values and agreement among echocardiographic measures of mechanical synchrony in healthy subjects are mostly lacking. The aims of this study were (1) to assess the ranges of normal values for 5 tissue Doppler imaging parameters, real-time 3-dimensional echocardiographic measures, and speckle-tracking measures of mechanical synchrony; (2) to evaluate interinstitutional variability; (3) to compare the ranges of normal values with those reported in previous research; and (4) to analyze the agreement among all parameters in the same healthy subject. Time to peak systolic velocity (Ts), the delay between Ts at the basal septal and lateral segments, peak velocity difference, strain derived by tissue Doppler imaging, Ts derived by tissue synchronization imaging, systolic synchrony index (SSI) derived by real-time 3-dimensional echocardiography, and longitudinal and radial strain derived by speckle tracking were prospectively collected and analyzed at 2 different institutions in 160 consecutive healthy subjects. The ranges of normal values, expressed as means +/- 2 SDs, were 30.32 +/- 29.36 ms for the SD of Ts, 15.51 +/- 99.88 ms for septal-lateral delay, 60.75 +/- 81.62 ms for peak velocity difference, 33.07 +/- 29.96 ms for tissue synchronization imaging, 34.16 +/- 23.26 ms for the SD of strain, 2.74 +/- 2.16% for SSI, 28.91 +/- 23.02 ms for the SD of longitudinal strain, and 10.4 +/- 6.31 ms for radial strain. There was large interinstitutional variability for all parameters. Three-dimensional SSI and radial strain were within the published upper range limit for healthy subjects. Ninety percent of healthy subjects were consistently classified to be synchronous by 1 parameter. With a composite index, more subjects than expected showed dyssynchrony (10% vs 2.5%). In conclusion, 3-dimensional SSI and radial strain were the most reproducible parameters and consistently discriminated normal healthy subjects from the cardiac resynchronization therapy volume responders.