Fabricated cantilever for AFM measurements and manipulations: Pre-stress analysis of stress fibers

► We measured strength of stress fibers in cells and stiffness of cells using specially shaped AFM cantilevers. ► The specially shaped cantilevers were fabricated by focused ion beam technique. ► Hook-shaped cantilevers can apply stronger force to stress fibers in the cell than conventional methods. ► The pre-stress of stress fiber in the cell was estimated by the force curve analysis. ► The combination of a microscoop cantilever and modified beads gave us high throughput measurement of cell stiffness.