Tissue Doppler imaging (TDI) is a relatively new echocardiographic technique that shows regional myocardial wall velocities. The aim of this study was to evaluate the potential value of acceleration mode TDI technique for the visualization of the origin of ventricular activation site using the model of right ventricular pacing. Twenty-seven patients with implanted permanent pacemakers were studied by acceleration mode TDI, 4 of these patients were pacemaker dependent. Parasternal and apical chamber views were recorded on video tape by using acceleration mode TDI technique during sinus rhythm with preserved atrioventricular conduction in 23 subjects who were not pacemaker-dependent, and also during right ventricular apical pacing in VVI mode in 27 subjects in whom pacing lower rate was increased if necessary. Fifty images recorded during sinus and pacing rhythm in cineloop were examined by two independent observers who were unaware of the rhythm patterns and by the same observer on two different occasions for localizing the site of onset of ventricular acceleration. The origin of ventricular activation during sinus rhythm started at basal septal part of the ventricle and during pacing started at apical part of the ventricle was considered as correct localizations. The origin of ventricular depolarization was correctly localized for 46 of 50 images (92%) and 44 of 50 images (88%) by the first and the second observers, respectively. Concordant results between observers appeared in 48 of 50 (96%) of images. The diagnostic accuracy of the concordant results was 44 of 48 (91.6%) images. The kappa for interobserver variability was 0.77 (p < 0.001), and for intraobserver variability was 0.64 (p < 0.001) and 0.63 (p < 0.001) for the first and the second observers, respectively. These results suggest that acceleration mode TDI can be used to detect the initial ventricular excited position and seems to have a potential value for localizing of the origin of normal or abnormal myocardial depolarization.