INTRODUCTION: Many cardiovascular diseases are closely associated with the mechanical properties of arterial wall and hemodynamic parameters. Simultaneous measurements of the arterial strain and flow pattern may aid diagnosis of cardiovascular diseases and may be useful to study fluid-structure interaction between blood and vessel. This paper proposes a 2D non-invasive ultrasonic method to simultaneously measure arterial strain and flow pattern with sub-pixel accuracy. MATERIALS AND METHODS: The method uses a multiple iterative algorithm to estimate the geometrical transformations of arterial wall and high-velocity gradient flows simultaneously. The accuracy of the method was validated by an in vitro arterial phantom and in vivo common carotid arteries (CCAs) of 12 mice using a Sonix RP (10 MHz) and a VisualSonics Vevo 2100 (30 MHz) ultrasound imaging system, respectively. RESULTS: For the arterial phantom, the calculated elasticity modulus from the strain profile shows good agreement with the mechanical testing value, deviating no more than 9.3%. The calculated flow velocity agrees well with the value obtained from the rotameter, deviating only 4.3%. For the CCAs of mice, good agreement is found between the calculated flow velocity and the measured value by ultrasound Doppler. The mean elasticity modulus of CCAs is 134.62 ± 54.3 kPa, which is in accordance with published data. CONCLUSION: The proposed method is capable of measuring the arterial wall strain and flow velocity pattern. This may be clinically useful for early detecting and monitoring cardiovascular diseases and may provide an essential tool in modelling the fluid-structure interaction between the blood and blood vessel.