Directed cell migration is fundamental to both physiological and pathophysiological processes such as embryogenesis, wound healing, and cancer metastasis. A complex series of events are required for directional cell migration, which is initiated by a migration-promoting or chemotactic stimulus, resulting in cellular polarization and entry into a cyclical pattern of leading edge protrusion, adhesion, and retraction of the trailing edge allowing cell movement. Heat shock proteins such as Hsp27, Hsp90, alphaB-crystallin, as well as heat shock transcription factors, are important players in both physiological and pathophysiological cell migration. A variety of techniques are currently available to assess cell migration, and among the most commonly utilized are those that employ a two-chamber methodology, such as that developed by Stephen Boyden in the early 1960s. Herein, we describe the use of a multiwell microchemotaxis migration assay that has the advantages of being reusable, inexpensive, highly reproducible, and requiring only a small volume of reagents.