In situ determination of exerted forces in magnetic pulling cytometry.

Localized application of exogenous forces on soft biomaterials and cells is often essential for the study of their response to external mechanical stimuli. Magnetic means of applying forces, particularly those based on permanent magnets and magnetic beads coupled to substrates or cells provide an accessible means of exerting forces of appropriate magnitude. The amount of force exerted, however, is often inferred from calibration performed ex situ, with typically similar but different magnetic beads. Here, we construct a simple magnetic tweezer by coupling a pencil-shaped stainless-steel probe to permanent neodymium magnets using a 3D printed adapter. We then demonstrate the in situ determination of magnetic bead pulling forces on a super-paramagnetic micro-bead coupled to a soft substrate using traction force microscopy. We determine the force exerted on the magnetic bead by the magnet probe and thus exerted by the magnetic bead on the soft polyacrylamide substrate as a function of the distance between the probe tip and the magnetic bead. We also show that we can determine the force exerted on a magnetic bead coupled to a cell by the changes in the traction force exerted by the cell on the soft substrate beneath. We thus demonstrate that forces of nanonewton magnitude can be locally exerted on soft substrates or cells and simultaneously determined using traction force microscopy. Application of this method for the in situ measurement of localized exogenous forces exerted on cells can also enable dissection of cellular force transmission pathways. (C) 2019 Author(s).