Image analysis of cell deposited material at cell periphery of live cells. A) Three time-lapse images of cell spreading on a 500 μm × 500 μm fibronectin patterned substrate separated by PEG that has been pre-incubated with serum medium exposure overnight. The times indicate 0 min, 30 min and 24 h after addition of cells to the chamber. Each image is overlaid with contour outlines depicting the distance-to-cell-edge coverage analysis routine (described in text) shown here at 20 μm from cell edge. For the 0 min starting time frame, the distance-to-cell-edge analysis routine used is the same as that for the 30 min time frame. B) The image extracted coverage versus distance-to-cell-edge data is displayed for the image at each time point. The starting mass/area is scaled to begin at 0 ng/cm2. The early time points (0 min, 30 min) show relatively even spatial distribution of surface coverage. The 24 h time point shows a mass distribution of putative cell-deposited material that occurs proximal to cell edge. Error bars represent the standard deviation from 4 different sample regions.