Figure 3.

The topographical method labels and quantifies invading cells when applied to endothelial layers. (A) The topographical method is applied to OOCs with different endothelial cells and visualized in 2-D, with x and y position shown on the x and y axes and z-height shown using a gradual color scale. The surfaces formed from different endothelial cells demonstrate different contour patterns. The HUVEC endothelial surface (left) shows regular, wave-like contours of a similar height across the surface. The HIMEC endothelial surface (right) demonstrates irregular contours with occasional endothelial objects sitting above or below the center surface. (B) 3-D reconstructions of endothelial surfaces formed from HUVECs (left) or HIMECs (right) showing the boundary used to label tumor cell objects as invading or non-invading. The tumor cells are labeled blue. (C) Numbers of invaded GFP+ tumor cells as measured using the ChICA tool. CRC OOCs with either HUVECs (left) or HIMECs (right) in the endothelial compartment were imaged on day 0 and day 6 of the experiment and invaded cells were identified based on the relative position to the endothelial surface.