Chemotaxis is a dynamic process of cell movement, which represents an important class of cell-cell communication through chemical signaling and plays crucial roles in many physiological and pathological processes. Besides the Boyden chamber assay which is common to study the cellular chemotaxis, microfluidic chamber assay has been gaining popularity for the macro-scale chemotaxis assays in real space and time with the disadvantages of generating a long-term stable and controllable gradient of soluble factors that can be continuously monitored over time.