PURPOSE: In this paper, a method for the estimation of arterial hemodynamic flow from x-ray video densitometry data is proposed and validated using an in vitro setup. METHODS: The method is based on the acquisition of three-dimensional rotational angiography and digital subtraction angiography sequences. A modest contrast injection rate (between 1 and 4 ml/s) leads to a contrast density that is modulated by the cardiac cycle, which can be measured in the x-ray signal. An optical flow based approach is used to estimate the blood flow velocities from the cyclic phases in the x-ray signal. RESULTS: The authors have validated this method in vitro, and present three clinical cases. The in vitro experiments compared the x-ray video densitometry results with the gold standard delivered by a flow meter. Linear correlation analysis and regression fitting showed that the ideal slope of 1 and intercept of 0 were contained within the 95 percentile confidence interval. The results show that a frame rate higher than 50 Hz allows measuring flows in the range of 2 ml/s to 6 ml/s within an accuracy of 5%. CONCLUSIONS: The in vitro and clinical results indicate that it is feasible to estimate blood flow in routine interventional procedures. The availability of an x-ray based method for quantitative flow estimation is particularly clinically useful for intra-cranial applications, where other methods, such as ultrasound Doppler, are not available.