Development of in vivo sonoporation method using short-pulsed ultrasound with microbubbles adjacent to cells.

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The final goal of this study was to develop a safe and efficient method for in vivo sonoporation that uses short-pulsed ultrasound in the presence of microbubbles adjacent to cells. A light microscopy system was developed for time-lapse observation of changes in cells generated during sonoporation and used to study methods to improve efficiency of the sonoporation. Optical tweezers were included in the system to control the position and extent of membrane damage by changing bubble size and position. Cell membrane damage and repair, which occur in a period shorter than 20 minutes, were observed using the system and their mechanisms were investigated. The observation chamber was improved to enable observation of long-term cell changes initiated by sonoporation, and the period of time-lapse observation was extended up to 20 hours. Two types of functional bubbles, tissue-targeting bubbles and drug-loaded bubbles, were developed, and their effectiveness for improving the efficacy of the proposed sonoporation method was confirmed.