Objective To investigate the effects of fluid shear stress on rolling adhesion of neutrophils on immobilized platelets under flows. Methods Experiments were performed at the parallel plate flow chamber. Platelets were adhered to the functionalized flow chamber bottom which were coated with vWF-A1 first, and then washed with PBS under wall shear stress (WSS) of 1 Pa for different time (0 min, 2.5 min, 7.5 min). A high-speed camera was used to observe and record the rolling adhesion events of neutrophils on immobilized platelets under 50 mPa WSS, and the adhesion parameters such as the number of adhesion events, the tether lifetime of cells and rolling velocity. Results Neutrophils could specifically bind to the immobilized platelets on vWF-A1-coated bottom of the flow chamber. Mechanical stimulation on immobilized platelets had no effects on the tether lifetime of neutrophils on the platelets, but up-regulated the adhesive ratio of neutrophils on the platelets and slowed down the rolling of neutrophils on the platelets. Conclusions Mechanical stimulation on the immobilized platelets will significantly make the circulating neutrophils to be captured easily and promote the rolling adhesion of neutrophils on platelets.