We describe a new approach for controlling cell motility on a material surface. Transparent, photosensitive polyimide materials were used to fabricate physical structures on glass; cell motility was then followed over time using optical microscopy. Arrays of pillars and holes with 2 micron square, 4-microm height (or depth) separated by 10 microm were successfully patterned using photolithography. Neutrophils attached and spread on the smooth glass surface and surfaces with pillars. In contrast, cells were rounded and did not adhere to either smooth polyimide film or films with holes. The migration of neutrophils was much faster on holes than on polyimide surface, but it was significantly slower on pillars than on glass. These results suggest that physical patterning may be an effective tool to manipulate cell migration in the design of biomaterials for tissue engineering.