OBJECTIVE Two-dimensional (2D) strain imaging has been established as a reliable and reproducible technique for the assessment of left and right ventricular function using transthoracic echocardiography (TTE). However, the reproducibility of transesophageal echocardiographic (TEE) 2D strain imaging and the agreement with TTE 2D strain imaging remains unclear. In the present study, the authors studied the reproducibility of TEE 2D strain imaging parameters. METHODS A comparative, observational clinical study. METHODS The echocardiography laboratory of the tertiary referral center. METHODS Healthy individuals with a suspected patent foramen ovale. METHODS None. RESULTS Thirty-four patients were included in the study. None of the patients had any structural cardiovascular disease. TTE and TEE images of the subjects were recorded and analyzed offline (EchoPAC 6.1; GE Vingmed Ultrasound AS, Horten, Norway). Longitudinal strain and strain rate measurements of the 4 chambers, the apical long axis, 2 chambers, and the right ventricle were obtained for each record of TTE and TEE. The mean age of the patients in this study was 36 ± 9.2 years. Bland-Altman analysis showed that there were generally good agreements between strain and strain rate measurements on TEE and TTE. The inter- and intraobserver agreement for TEE parameters was good. CONCLUSIONS Transesophageal 2D strain imaging is a reproducible method to measure ventricular function and has a good agreement with TTE 2D strain imaging.