A crucial task in inflammation research and inflammatory drug validation is leukocyte velocity data collection from microscopic video imagery. Since manual methods are bias-prone and extremely time consuming, automated tracking methods are required to compute cell velocities. However, an automated tracking method is of little practical use unless it is accompanied by a mechanism to validate the tracker output. In this paper, we propose a validation technique that accepts or rejects the output of automated tracking methods. The proposed method first generates a spatiotemporal image from the cell locations given by a tracking method; then, it segments the spatiotemporal image to detect the presence or absence of a leukocyte. For segmenting the spatiotemporal images, we employ an edge-direction sensitive nonlinear filter followed by an active contour based technique. The proposed nonlinear filter, the maximum absolute average directional derivative (MAADD), first computes the magnitude of the mean directional derivative over an oriented line segment and then chooses the maximum of all such values within a range of orientations of the line segment. The proposed active contour segmentation is obtained via growing contours controlled by a two-dimensional force field, which is constructed by imposing a Dirichlet boundary condition on the gradient vector flow (GVF) field equations. The performance of the proposed validation method is reported here for the outputs of three different tracking techniques: the method was successful in 97% of the trials using manual tracking, in 94% using correlation tracking and in 93% using active contour tracking.