A novel method for quantifying traction forces on hexagonal micropatterned protein features on deformable poly-dimethyl siloxane sheets.

Many methods exist for quantifying cellular traction forces, including traction force microscopy and microfabricated post arrays. However, these methodologies have limitations, including a requirement to remove cells to determine undeflected particle locations and the inability to quantify forces of cells with low cytoskeletal stiffness, respectively. Here we present a novel method of traction force quantification that eliminates both of these limitations. Through the use of a hexagonal pattern of microcontact-printed protein spots, a novel computational algorithm, and thin surfaces of polydimethyl siloxane (PDMS) blends, we demonstrate a system that: •quantifies cellular forces on a homogeneous surface that is stable and easily manufactured.•utilizes hexagonal patterns of protein spots and computational geometry to quantify cellular forces without need for cell removal.•quantifies cellular forces in cells with low cytoskeletal rigidity.