The evaluation of left ventricular diastolic function provides important information about hemodynamics and has prognostic implications for various cardiac diseases. In particular, left atrial (LA) volume is an increasingly significant prognostic biomarker for diastolic dysfunction. The aim of this study was to assess left ventricular diastolic function by measuring changes in LA volume using real-time 3-dimensional echocardiography. The 106 subjects were divided into 4 groups (normal, impaired relaxation, pseudonormal, and restrictive) on the basis of diastolic function, as assessed by transmitral flow patterns. LA volume was measured during a heart cycle using real-time 3-dimensional echocardiography. LA stroke volume (maximum LA volume - minimum LA volume) and the LA ejection fraction (LA stroke volume/maximum LA volume x 100) were calculated using Doppler imaging to assess their correlation with other parameters used to evaluate left ventricular diastolic function, including transmitral flow pattern and early diastolic mitral annular velocity (E'). LA volume indexed to body surface area was dilated in subjects with left ventricular diastolic dysfunction, whereas the LA ejection fraction was lower. The maximum LA volume, minimum LA volume, and LA ejection fraction were significantly different between each group, and each was significantly correlated with the ratio of early diastolic transmitral flow velocity (E) to E' (E/E'). The LA ejection fraction correlated best with E/E' (r = -0.68, p <0.0001). In conclusion, cyclic changes in LA volume could be measured using real-time 3-dimensional echocardiography, and measuring LA function with this method may be a viable alternative for the accurate assessment of left ventricular diastolic function.