Background: Limited number of studies evaluated cardiac diastolic function in infants of diabetic mothers using tissue Doppler imaging. The aim of this study was to evaluate diastolic parameters in full-term infants of diabetic mothers compared to healthy full-term neonates using both conventional echocardiography and tissue Doppler imaging. This study is a comparative study. Fifty consecutive infants of diabetic mothers (cases) in the first 3 days of life: 25 neonates with poor maternal glycemic control (Hemoglobin A1c > 7.5 g/dl) and 25 neonates with good maternal glycemic control (Hemoglobin A1c ≤ 7.5 g/dl). Thirty healthy full-term infants of non-diabetic mothers with age and sex matching were included as controls. The studied groups were assessed by conventional pulsed wave Doppler and tissue Doppler imaging. Results: Among pulsed wave Doppler parameters, cases had lower values than controls as regards mitral E velocity, mitral E/A ratio, tricuspid E velocity, and tricuspid E/A ratio, while neonates with poor maternal glycemic control had lower values than those with good maternal glycemic control as regards mitral E/A ratio, tricuspid E velocity, and tricuspid E/A ratio as well as higher mitral A velocity (denoting more diastolic dysfunction). Similarly, among tissue Doppler parameters, cases had lower values than controls as regards septal E' velocity, E'/A' ratio, left ventricular E' velocity, E'/A' ratio, and right ventricular E' velocity as well as higher septal A' velocity, left ventricular A' velocity, and right ventricular A' velocity, while neonates with poor maternal glycemic control had lower values than those with good maternal glycemic control as regards septal E' velocity, E'/A' ratio, left ventricular E' velocity, E'/A' ratio, and right ventricular E' velocity, as well as higher left ventricular A' velocity (denoting more diastolic dysfunction). Tissue Doppler was able to detect higher number of neonates with left ventricular diastolic dysfunction than conventional pulsed wave Doppler. Conclusions: Tissue Doppler imaging was found to be able to detect diastolic dysfunction early in infants of diabetic mothers specifically as regards the left ventricle. Tissue Doppler imaging should be considered an integral part of cardiac function assessment in infants of diabetic mothers.