Measuring surface geometry of adherent cells using oblique transillumination

We present a new technique for estimating cell surface geometry. A dish supporting adherent cells is observed using oblique transillumination and rotated in the horizontal plane using a stepping motor. The stage rotation-dependent movements of the start and end points of a shadow formed behind the illuminated cells uniquely determine the relative height differences between points along the cell surface. Thus, using custom-made apparatuses and living endothelial cells, we demonstrate that the combination of a rotating stage and oblique lighting allows for the evaluation of three-dimensional surface geometry of adherent cells. As compared to confocal microscopy and atomic force microscopy, which are commonly used for measuring cell surface geometry, this approach can be performed rapidly and is especially suitable for the observation of unstained cells over a large surface covering multiple cells at a time.