Boyden Chamber Invasion Assays
20,000 SCC9 cells were suspended in 200 ul of serum-free growth media and seeded inside a matrigel-coated transwell. 750 ul of growth media containing 10% FBS was added to the larger well. Cells were allowed to migrate through the pores to the other side of the membrane by incubating them at 37 degrees Celsius for 24 hours. After 24 hours of incubation, the transwells were washed in PBS, fixed in 100% Methanol for 2 minutes, and then stained in crystal violet solution for 20 minutes. Excess crystal violet was washed off by dipping the transwells in water. The transwells were then left to dry following which they were imaged in a brightfield microscope. A custom Image J script was used to count cells that migrated into the Matrigel coated side of the transwell.