Visualization of Ovarian Cancer Cell Invasion

Cancer cells were live stained with PKH67 cytotracker-green (Sigma-Aldrich, USA) before seeding into the device. After extravasation, the ovarian cancer cells were incubated at 37°C with continuous perfusion of medium in tumor chamber for 3 days. Invasion dynamics of live-stained cancer cells (green) into the chip ECM chamber hydrogel was visualized using fluorescence microscopy (ZEISS Axio Observer; LD Plan Neofluar 10×, numerical aperture 0.4). Snapshots were taken at every 12 hours of interval with an exposure time of 200 ms. Images were analyzed and processed using software ZEN 2.3 lite (ZEISS). Measurement of cancer cell invasion through the hydrogel was performed using cell counter plugins in ImageJ. The ECM invasion of cancer cells was calculated as the ratio of area occupied by invaded cancer cells in ECM chamber versus the total area of ECM chamber (55 (link)).