FIG. 9.

Optical DIC image sequence (7 days) of HPCFs plating directly on chamber glass bottom. On day 0, there are many rounded cells above the glass (as determined by their random motion). By day 1, the glass is coated to confluence by the cells that are randomly oriented. By day 3, there appears to be organized sheet motion of the layer of cells (upward in image). Debris on the culture surface is tracked (black circles) and appears to keep pace with cell movement. In addition to the whole culture sheet motion vertically, there was a general horizontal motion of individual cells, which was faster, variable, and became more marked at days 6 and 7. (Bars 100 μm; see Movie in Supplementary Data for dynamic images of the behavior). Note, occasionally, there are jumps in the movie sequence or shifts in the fluid over the cells; these change could be due to either inadvertent “bumps” of the instrument or failure to relocate after clearing of a bubble. DIC, differential contrast imaging; HPCF, human primary corneal fibroblast.