P-selectin expressed on the inflamed endothelial surface is a key mediator of the initial steps of inflammation. It has recently been shown that the local variation of endothelial P-selectin expression affects leukocyte recruitment in vivo. Yet, little is known about the dynamics of leukocyte adhesion a patterned P-selectin-surface in vitro. We investigated the influence of microcontact printing of P-selectin on a polystyrene surface in leukocyte recruitment using a parallel-plate flow chamber. Micro-stripe (100 µm × 10 mm) patterns of P-selectin were prepared using a PMDS stamp and isolated human neutrophils perfused over the surface to quantify the level of neutrophil recruitment. We first found a significant increase in cell rolling flux and a decrease in cell rolling velocity on the micropatterned P-selectin surface compared with a randomly adsorbed P-selectin-surface. However, it was determined using immunofluorescent labeling that the number of functional sites of P-selectin transferred by the microcontact printing was less than that by random adsorption. In addition, by using an anti-P-selectin antibody specific for the short consensus repeat domain, the immuno-labeling of the non-functional sites of P-selectin immobilized on a surface was substantially less on the microcontact printed surface. Taken together, these data indicate that the microcontact printing of selectin increases the transfer rate of the adhesion molecule on a surface in the functionally correct orientation and, consequently, improves the recruitment of leukocytes to the selectin surface under flow.