Background: Three-dimensional speckle-tracking echocardiography (3DSTE) is a relatively new noninvasive imaging modality with ability for simultaneous chamber quantifications and determination of valvular dimensions. The aim of the present retrospective cohort study was to determine normal reference values of 3DSTE-derived tricuspid annular (TA) dimensions and functional properties and to evaluate their age-and gender dependency. Methods: The present study comprised 156 healthy adult subjects, from which 28 cases were excluded due to inferior image quality, therefore the remaining group consisted of 128 cases with the mean age of 35.4±12.5 years (72 males). The subject population was further divided into the following categories: 18-29 years (n=57; mean age: 25.2±2.8 years, 51 males), 30-39 years (n=29; mean age: 34.1±2.5 years, 31 males), 40-49 years (n=17; mean age: 44.1±3.2 years, 11 males) and ≥50 years of age (n=25, mean age: 59.2±6.4 years, 14 males).Results: End-diastolic TA diameter (2.2±0.3 vs. 2.5±0.3 cm, P<0.05), area (7.1±1.3 vs. 8.1±1.7 cm 2 , P<0.05) and perimeter (10.3±0.9 vs. 11.0±1.2 cm, P<0.05) were lower in females than in males in the 18-29 yearold group and TA area (6.1±0.8 vs. 8.0±1.2 cm 2 , P<0.05) and TA perimeter (9.7±0.8 vs. 11.0±0.9 cm, P<0.05) were lower in females than in males in the 40-49 year-old group. End-systolic TA diameter were lower in females than in males in the 18-29 year-old group (1.8±0.2 vs. 1.9±0.3 cm, P<0.05) and TA area (4.7±0.3 vs. 6.3±1.2 cm 2 , P<0.05) and TA perimeter (8.6±0.4 vs. 9.5±0.9 cm, P<0.05) were lower in females than in males in the 40-49 year-old group. TA fractional area change was found to be reduced in the 40-49 year-old group as compared to the 30-39 year-old group (21.7%±8.7% vs. 29.2%±10.0%, P<0.05).Conclusions: 3DSTE is a novel method for non-invasive assessment of TA dimensions and functional properties. Results highlight the importance of age-and gender-specific reference values in case of TA diameter, area and perimeter and calculated functional features respecting the cardiac cycle.