Vascular endothelial cells subjected to fluid shear stress change their shape from polygonal to ellipsoidal and become uniformly oriented with the flow. In order to study the mechanisms of this response, we have measured the relaxation of bovine aortic endothelial cells that were grown on glass coverslips and exposed to fluid shear stress for 72 hours. An image analysis system was developed to quantify the cell shape relaxation that occurs following the cessation of shear stress. This method provides two different quantitative measures of relaxation: the loss of elongated shape by the cells and the change in cell direction with time. After equilibration to a fluid shear stress level of 8 dynes/cm 2, cells immersed in static medium relax their shape in about 20 hours. After 72 hours in this static condition, the cell elongation is comparable to that of unstressed control cells but vestiges remain of the original orientation in the flow direction. This relaxation process contributes to our understanding of the response of vascular endothelium to fluid shear stress.