The deformability of human platelets has been evaluated by micropipette aspiration. Control discoid platelets were about ten times as resistant to deformation in the micropipette as red blood cells. Under a constant negative pressure of 10 cm H2O, control platelets developed extension lengths of 0.74 +/- 0.1 micron. Prior treatment with vincristine, colchicine, or low temperature, all of which remove platelet microtubules, was associated with marked increases in lengths of aspirated segments. Taxol or heavy water, which stabilize microtubules, prevented the increased deformability caused by agents that dissolve microtubules. Cytochalasin B, an agent that inhibits assembly of actin microfilaments, also caused an increase in lengths of aspirated segments that could not be prevented by taxol. Vincristine and cytochalasin B, together, caused a greater increase in deformability than either agent alone. These results indicate important roles for microtubules and microfilaments in platelet deformability.