Microbubbles have potential for applications as drug and gene delivery systems in which the release of a substance is triggered by an ultrasonic pulse. In this paper, we discuss the adsorption and desorption of a film of phospholipid on the surface of a single microbubble under ultrasound irradiation. Our optical observation system consisted of a high-speed camera, a laser Doppler vibrometer, and an ultrasound cell; 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) was used as the surfactant. The adsorption of the DMPC molecules onto the surface of the bubble was evaluated by measuring the contact angle between the bubble and a glass plate. A decrease of the contact angle of the bubble indicates desorption of the DMPC molecules from the bubble surface into the surrounding aqueous solution. The amount of DMPC molecules adsorbed on the bubble's surface is shown to decrease over time after bubble generation. The type and intensity of the pulsed ultrasound waves were varied so as mimic ultrasound triggered drug release. Increasing the number of cycles and the amplitude of the sound pressure of the pulsed ultrasound yielded a greater increase of the contact angle. We also measured the radial vibrations of the microbubbles in the ultrasound field. The vibrational characteristics of the microbubbles and the desorption characteristics of the DMPC molecules showed the same variation; namely, a greater sound pressure amplitude induced greater vibrational displacement and a larger amount of molecular desorption under resonance conditions. These results support the possibility of controlling drug release with pulsed ultrasound in a microbubble-based drug delivery system.