Changes in traction forces in cells after force application. (A) Immunofluorescent micrograph of cell A, force-stimulated with a magnetic post, and cell B, unstimulated control. Labeling for actin (cyan), nuclei (blue), and PDMS (brown) were performed immediately after traction force video observation. Posts of interest are marked with colored circles and labeled according to which cell they were attached to (Ai or Bi) or background (Bkg). (B) Plot of displacement and force versus time for all posts for cell A (light red). A subset of posts of interest are designated (AI, red; BI, green; magnetic post A2, yellow; and background post, blue). Onset of force stimulation is indicated by dashed line (t = 0). The force reported by the deflection also reflects cellular traction forces, except for A2, which has an additional magnetic force component of 1.3 nN introduced at t = 0. (C) Plot of average strain energy per post (u) versus time for force-stimulated cells. Each cell is categorized by their response: sudden (red), gradual (purple), and no apparent (black) response. The cell marked with asterisk (*) has large strain energy and is shown with a scaling reduction of 3. (D) Plot of u versus time for unstimulated control cells (green) and background posts for all experiments (blue). (B–D) Error bars for all graphs indicate uncertainty in analysis. (E) Spatial plots of change in strain energy (Δui) in posts immediately before and after force application for stimulated cells that have sudden responses. Posts underneath each cell are shown in each plot (+). For each spatial plot, the direction of applied field (B = 0.2 T) is upward. For cell A, FMag = 1.3 nN, and for cell C, FMag = 3.3 nN. For posts with Δui outside the range of the scale, they are colored as the nearest extrema. (F) Spatial plots of Δui in posts immediately before and 10 min after for stimulated cells responding with gradual changes in energy. For cell D, FMag, top = 5.4 nN and FMag, bottom = 6.2 nN, and for cell E, FMag = 3.9 nN. (G) Spatial plots of control cells comparing Δui immediately before and after force application.