Probing the cell peripheral movements by optical trapping technique.

Abstract

Swiss 3T3 fibroblasts cultured on a poly-L-lysine-coated coverslip was stimulated with 0.5 micro M phorbol myristate acetate, and the movements of the peripheral membranes were probed with a 1- micro m polystyrene bead held in an optical trap. The bead brought into contact with the cell edge occasionally moved away from and returned to the original position. The movement ranged over 100 nm and occurred mainly in one direction, suggesting that the protruding cell membrane pushed the bead. The maximum velocities derived from individual pairs of protrusive and withdrawal movements exhibited a correlation, which is consistent with the previous reports. Acceleration and deceleration occurred both in the protrusive and withdrawal phases, indicating that the movements were regulated. Movement of the membrane occurred frequently with an ensemble-averaged maximum speed of 23 nm/s at the trap stiffness of 0.024 pN/nm, but it was strongly suppressed when the trap stiffness was increased to 0.090 pN/nm. Correlation of the protrusive and withdrawal velocities and the acceleration and deceleration both in the protrusive and withdrawal phases can be explained by the involvement of myosin motor at least in the withdrawal process. However, the fact that the movements were suppressed at higher trap stiffness implies a stochastic nature in the creation of the gap between the peripheral cell membrane and the actin network underlying it.