New Findings What is the central question of this study? The aim was to investigate whether complementary assessment of non‐invasive ultrasound imaging together with closed chest‐derived intracardiac pressure–volume catheterization is applicable to mice for an in‐depth characterization of right ventricular (RV) function even upon maintained pressure overload. What is the main finding and its importance? Characterization of RV function by the complementary use of echocardiographic imaging together with pressure–volume catheterization reveals ventricular–arterial decoupling upon maintained pressure overload, where RV systolic function correlates with ventricular–arterial coupling rather than contractility, whereas diastolic function correlates well with RV diastolic pressure. This combined approach allows us to phenotype RV function and dysfunction better in genetically modified and/or pharmacologically treated mice. Assessment of right ventricular (RV) function in rodents is a challenge because of the complex RV anatomy and structure. To date, the best characterization of RV function has been achieved by accurate cardiovascular phenotyping, involving a combination of non‐invasive imaging and intracardiac pressure–volume measurements. We sought to investigate the feasibility of two complementary phenotyping techniques for the evaluation of RV function in an experimental mouse model of sustained RV pressure overload. Mice underwent either sham surgery (n = 5) or pulmonary artery banding (n = 8) to induce isolated RV pressure overload. After 3 weeks, indices of RV function were assessed by echocardiography (Vevo2100) and closed chest‐derived invasive pressure–volume measurements (PVR‐1030). Pulmonary artery banding resulted in RV hypertrophy and dilatation accompanied by systolic and diastolic dysfunction. Invasive RV haemodynamic measurements demonstrated an increased end‐systolic elastance and arterial elastance after pulmonary artery banding compared with sham operation, resulting in ventricular–arterial decoupling. Regression analysis revealed that tricuspid annular plane systolic excursion is correlated with ventricular–arterial coupling (r² = 0.77, P = 0.002) rather than with RV contractility (r² = −0.61, P = 0.07). Furthermore, the isovolumic relaxation time to ECG‐derived R–R interval and the ratio of the early diastolic peak velocity measured by pulsed wave Doppler to the early diastolic peak obtained during tissue Doppler imaging correlate well with RV end‐diastolic pressure (r² = 0.87, P = 0.0001 and r² = 0.82, P = 0.0009, respectively). Commonly used indices of systolic RV function are associated with RV–arterial coupling rather than contractility, whereas diastolic indices well correlate with end‐diastolic pressure when there is maintained pressure overload.