Objectives: Echocardiographic assessment of Left ventricular systolic function is traditionally being performed by estimation of fractional shortening and Ejection fraction. Speckle tracking echocardiography (STE) is a promising tool for assessment of myocardial function. The aim of this study is to evaluate the global longitudinal strain (GLS) using 2D-STE in healthy neonates to establish normal reference ranges. Method: it is a retrospective study through an analysis of transthoracic echocardiogram of normal healthy neonates. We enrolled all neonates in our institution from 1 st January 2021 to 28 th February 2021. 2-D STE was used to assess left ventricular global longitudinal strain from the apical views. Results: 185 neonates were enrolled. Mean value for left ventricle GLS (%) was -19.9 ± 1.2, GLS-derived ejection fraction (%) was 60.0 ± 2.7; while the left ventricle ejection fraction by biplane Simpson’s method (%) was 61.0 ± 3. There is a good positive correlation between the Left Ventricle EF by biplane Simpson’s method and EF by 2-D STE, which was statistically significant ( r = 0.294,  n = 102,  p=0.003). Apical 4-chamber longitudinal strain and strain derived EF is significantly correlated with GLS and bi-plan EF respectively. Conclusion: 2-STE is feasible technique for analyzing newborn myocardial systolic function. The normal range of GLS in neonates is not much different than reported for the pediatric. There is a good positive correlation between the Left Ventricle EF by 2-D STE and EF by biplane method.