Directional cellular migrations as a chemotactic response to spatially inhomogeneous growth factor stimulation play an important role in establishing physiological mechanisms and pathological events in cells. We developed epidermal growth factor (EGF)-immobilized microbeads by photoreaction and evaluated its local stimulatory effects on the dynamic chemotactic motility of fibroblasts. The local stimulation resulted in global activation of ERK 1/2 and directionality of cellular migration. The cellular migration by stimulation using 3-mu m diameter EGF-immobilized microbeads persisted for a longer time, were involved a wider field and their number were further increased with stimulation. This effective technique allows cellular migration and biochemical analyses that will help elucidate the mechanisms involved in signal transduction by spatially inhomogeneous stimulation of the growth factor. (C) 2013 Elsevier Ltd. All rights reserved.