Background: Blood sampling from arterial lines is a frequent event in anesthesia and critical care. To avoid clot formation, both the stopcock outlet and the cannula must be flushed after sampling. We investigated in a bench experiment whether fluid flow through the cannula is affected by the sequence of flushing procedures. Methods: Continuity of fluid delivery from a vascular cannula was gravimetrically determined using two different flushing techniques with either a syringe pump flush system or a bag flush system. The procedures comprised first flushing the stopcock towards the cannula and then towards the stopcock sampling outlet or the reverse order. Experiments were repeated in triplicate and two sets for each flushing system at hydrostatic pressures of 37 mm Hg and 74 mm Hg. Results : The main finding of the study was that flushing the stopcock towards the outlet after flushing the cannula resulted in considerable retrograde aspiration volumes and zero flow times, in particular in combination with syringe pump flush systems.