Aim To detect the accuracy of real-time threedimensional echocardiography (RT3DE) and two-dimensional echocardiography (2DE) for tricuspid annulus (TA) assessment compared with magnetic resonance imaging (MRI). Methods Thirty patients (mean age 34 ± 13 years, 60% males) in sinus rhythm were examined by MRI, RT3DE, and 2DE for TA assessment. End-diastolic and end-systolic TA diameter (TAD) and TA fractional shortening (TAFS) were measured by RT3DE, 2DE, and MRI. End-diastolic and end-systolic TA area (TAA) and TA fractional area changes (TAFAC) were measured by RT3DE and MRI. End-diastolic and end-systolic right ventricular (RV) volumes and ejection fraction (RV-EF) were measured by MRI. Results The TA was clearly delineated in all patients and visualized as an oval-shaped by RT3DE and MRI. There was a good correlation between TAD MRI and TAD 3D (r = 0.75, P = 0.001), while TAD 2D was fairly correlated with TAD 3D and TAD MRI (r = 0.5, P = 0.01 for both). There were no significant differences between RT3DE and MRI in TAD, TAA, TAFS, and TAFAC measurements, while TAD 2D and TAFS 2D were significantly underestimated (P < 0.001). TAFS 2D was not correlated with RV-EF, while TAFS 3D and TAFAC 3D were fairly correlated with RV-EF (r = 0.49, P = 0.01, and r = 0.47, P = 0.02 respectively). Conclusion RT3DE helps in accurate assessment of TA comparable to MRI and may have an important implication in the TV surgical decisionmaking processes. RT3DE analysis of TA function could be used as a marker of RV function.