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.