Introduction: The present study was designed to define normal reference values of threedimensional speckle-tracking echocardiography (3DSTE)-derived left atrial (LA) volumes, stroke volumes (SVs), and emptying fractions (EFs) with regard to the cardiac cycle. Methods: The present study involved 256 healthy adult subjects in sinus rhythm who underwent complete two-dimensional Doppler echocardiography and 3DSTE at the same time. However, due to inferior image quality, 87 subjects have been excluded. The remaining population sample comprised of 169 patients who gave informed consent to participate in the study. Results: While systolic maximum LA volume and early diastolic preatrial contraction LA volume did not change over age decades, late-diastolic minimum LA volume decreased and was lowest in subjects aged 40 to 49 years then increased after 50 years. Total atrial EF increased over age decades with a reduction after 50 years. Passive atrial EF showed a significant continuous increase over age decades. Active atrial EF did not change in younger ages and was the highest between ages 40 and 49 years with a significant impairment after 50 years. Conclusions: This study shows the age-and gender-dependency of normal values of 3DSTE-derived LA volumes, stroke volumes, and emptying fractions with regard to the cardiac cycle in healthy adult subjects.