ABSTRACT Objective To explore the feasibility of measuring the mobility and contraction velocity of bilateral diaphragm by anatomical M-mode ultrasound, so as to provide basis for the diagnosis of diaphragm motor dysfunction. Methods A total of 118 healthy adults were selected to measure the movement and inspiratory time of bilateral diaphragm during quiet breathing with anatomical M-mode ultrasound by the same examiner, and the contraction velocity of diaphragm was calculated. Intraclass correlation coefficient（ICC) was used to evaluate the repeatability of ultrasound parameters, and to analyze the relationship between diaphragm movement and sex, age, smoking history, body mass index (BMI). Results The normal reference range of diaphragm movement parameters: mobility (9~23) mm, inspiratory time (973~1483) ms, systolic velocity (0.82-1.87) cm/s, there was no significant difference in diaphragm movement parameters; anatomical M-mode ultrasound measured diaphragm mobility and inspiratory time ICC values were more than 0.9. There was no significant correlation between diaphragm movement and sex, age, smoking history and BMI. Conclusions Anatomical M-mode ultrasound can accurately obtain the mobility of bilateral diaphragm, and has good repeatability, which overcomes the deficiency that the traditional M-mode ultrasound can only obtain the mobility of right diaphragm. It can be widely used in clinic as a non-invasive and non-radiation imaging method for monitoring diaphragm movement.