Abstract Funding Acknowledgements Philips Medical Systems Background Continuous efforts of EACVI-ASE strain standardization task force make inter-vendor variability of left ventricular (LV) global longitudinal strain (GLS) lower. However, observer variability is still a major source of measurement inconsistency. The adoption of fully automated 2D strain software has a potential to eliminate this problem. Purpose We aimed to validate the usefulness of novel fully automated LV, left atrial (LA), and right ventricular (RV) 2D speckle tracking software packages. Methods We retrospectively selected 91 healthy subjects who had undergone 2D echocardiography using a specific ultrasound machine (EPIQ, Philips Medical Systems). Standard apical 4-chamber, 2-chamber, and long-axis views which encompassed whole part of both left ventricle and left atrium and RV focused view were acquired in all subjects. Novel fully automated speckle tracking software packages (AutoStrain, QLAB 13.0) were used for measuring GLS on the three apical views, LA longitudinal strain (LALS) on the apical-4-chamber view, and RV free wall longitudinal strain (RVfwLS) on the RV-focused view. Image quality was assessed by 3-point scales (good, fair, and poor). Endocardial border detection was also classified into 3 groups (adequate, poor, and no detection). Endocardial border was manually adjusted, when required. Results of fully automated analysis and results with fully automated analysis + manual editing were compared. Results Mean age was 37 ± 13 years, and 61 subjects were men. The image quality was categorized more than "fair" in 80% of subjects. The software did not recognize LA border in 3 cases and RV border in 4 cases. Thus, the feasibility of fully automated LV, LA, and RV analysis were 100%, 97%, and 96%, respectively. LVGLS, LALS, and RVfwLS using fully automated approach were 19.7 ± 2.3%, 45.5 ± 11.6%, and 25.7 ± 5.6%, respectively. Manual correction was required in all cases. LVGLS, LALS, and RVfwLS after the manual editing were 18.9 ± 2.1%, 44.0 ± 10.4%, and 26.6 ± 6.4%. The intraclass correlation coefficient (ICC) between the two methods were 0.86, 0.72, and 0.74. The fully automated analysis took 22 ± 2 sec, 11 ± 1 sec, and 10 ± 1 sec for the measurements of LVGLS, LALS, and RVfwLS. The corresponding values with manual editing were 98 ± 18 sec, 53 ± 12 sec, and 52 ± 12 sec, respectively. Inter-observer ICCs of LVGLS, LALS, and RVfwLS with fully automated approach were all 1.0, but corresponding values after the manual editing were 0.82, 0.74, and 0.79, respectively. Conclusions Novel fully automated 2D speckle tracking software packages provide LVGLS, LALS, and RVfwLS within one minute, and these values were well correlated with the corresponding values after the manual editing, especially for LVGLS. This is a time-saving approach for longitudinal strain analysis in the three cardiac chambers. Further studies should be required to validate their potential utility in clinical setting.