(a) CFD predictions of axial flow velocity (upper) and shear stress (lower) in the flow chamber at 10 µm from the bottom of the flow chamber. Flow moves from right (inlet) to left (outlet). (b) 3D images of flow fields (red arrows indicate microsphere displacements) around cells (green). Three dimensional confocal image stacks were analyzed to quantify the flow fields with respect to distance from the substrate and cell density. (c) Confocal images close to the basal surface, approximately 2 µm from the substrate on which cells are seeded (A,D,G), 5 µm from the substrate (B,E,H), and 10 µm from the substrate (C,F,I). Cells are labeled with calcein green, microspheres exhibit red fluorescence, and white arrows indicated microsphere displacements in 990 milliseconds. Green, red, and white indicate cells, microspheres, and displacements, respectively.