Physical factors like stiffness and surface morphology of the substrata are thought to influence cell behaviors. How the cell motility at early attachment influenced by the substrates was not investigated yet. The purpose of this study is to measure the cell motility under different stiffness and pattern of the substrata. PDMS groups moved significant faster than glass due to smaller stiffness. In addition, over 80% of cells on long groove PDMS moved along the groove, indicating long groove was the best surface morphology to control cell directional movement.