Directed crawling and shape generation of white blood cells on a surface has been extensively investigated as a simplified in vitro system for modelling leukocyte migration in the immunological responses. Surface topographical anisotropy and mechanical stiffness are known to be main physical cues that affect the leukocyte responses, however, their combined effects have been poorly understood particularly for neutrophils, the fastest reacting leukocytes against infections and wounds. In this presentation, we elucidate the transitory process of the global shape modulation during neutrophil crawling on a microgrooved surface covered with adsorbed extracellular matrix proteins, under varying the substrate elasticity. We quantitatively demonstrate that the synergy between the substrate topography and stiffness may drastically influence the temporal morphology evolution of a migrating neutrophil on a 2D scaffold. The spatial and mechanical control of the substrates significantly affects the cell shape-translocation relationships through contact guidance and stiffness feedback.