Cell exclusion zone assay is especially appropriate for studying cell migration on an uncompromised surface uncoupled from contributions of cell damage and permeabilization. To avoid damaging cells and matrix, cell-free areas are created in cell exclusion zone assay by that cells are seeded around removable silicone stoppers which exclude cells from attaching to a central zone. The cell density is adjusted to form a fully confluent. After cell adhesion, the stoppers are removed to reveal a circular cell-free area, wherein the cells will migrate. Then, upon the appropriate staining with fluorescent dyes and microscopically visualizing the monolayers, the migration rate is quantified by counting the cells which have migrated to the circular cell-free area.