Here we will simultaneously measure internal cell forces, external cell contractility, cell-cell forces, and cell movement. However, how this increased external cell contractility orchestrates internal mechanical changes during EMT to promote migration is not well understood. Our lab and others have previously shown that as epithelial cells acquire more migratory and invasive mesenchymal characteristics, they apply more forces on their surroundings and do more work. Cancer cells are known to undergo various biomechanical changes during this transition, yet a comprehensive and cohesive quantification of these changes is lacking. This biophysical change is believed to contribute to the increased tumor cell motility in cancer metastasis.
During EMT, tightly packed epithelial cells become more invasive and motile mesenchymal cells. The Epithelial-to-Mesenchymal Transition (EMT) is a key transformation in cancer metastasis. Now we want to develop PaCS as a high throughput technology that measures more than 1000 cells at once. We already validated this approach by comparing this Pattern-based Contractility Screening (PaCS) to conventional bead-displacement TFM and show quantitative agreement between the methodologies. In this technique, we confine the cells on fluorescent adhesive protein micropatterns of a known area on compliant silicone substrates and use the cell deformed pattern area to calculate cell contractile work. Here we introduce a reference-free technique to measure cell contractile work in real-time, with basic substrate fabrication methodologies, simple imaging, high-throughput potential and analysis with the availability of the cells for post-processing. While TFM platforms have enabled diverse discoveries, their implementation remains limited in part due to various constraints, such as complex substrate fabrications, the need to detach cells to measure null force images, followed by complex imaging and analysis, limited cell number, and the unavailability of cells for post-processing. Traction Force Microscopy (TFM) has emerged as a standard broadly applicable methodology to measure cell contractility and its role in cell behavior. The sensing and generation of cellular forces are essential aspects of life.
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