Umebayashi Yukihiro, Miyamoto Yuuichi, Wakita Masayoshi, Kobayashi Akiko, Nishisaka Tsuyoshi
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Tatsunokuchi, Nomi-gun, Ishikawa 923-1292.
J Biochem. 2003 Aug;134(2):219-24. doi: 10.1093/jb/mvg132.
In this report, we describe a laser-latex combination system that enables membrane-impermeable molecules to penetrate cell membranes. Laser light (Q-switched Nd:YAG laser, 532.5 nm) was used to irradiate a mixture of commercial latex particles (blue dyed, 1 micro m in diameter) and mouse fibrosarcoma (Meth-A) cells. After irradiation, membrane permeability was evaluated by flow cytometric assaying using propidium iodide (PI) and fluorescein diacetate (FDA). The proportion of permeabilized-resealed cells was affected by changes in the light intensity (approximately 780 mW/cm(2)), the irradiation time (approximately 240 s), and/or the particle concentration (approximately 10(9) particles/ml). The permeability persisted up to 20 min after light irradiation. Near the sites of individual particles, the permeability of the cell membrane is modified, probably due to localized temperature changes. These results suggest that this laser-induced permeabilization strategy constitutes a new means of delivering exogenous materials into living cells.
在本报告中,我们描述了一种激光-乳胶组合系统,该系统能使膜不可渗透的分子穿透细胞膜。使用激光(调Q Nd:YAG激光,532.5纳米)照射商业乳胶颗粒(蓝色染色,直径1微米)与小鼠纤维肉瘤(Meth-A)细胞的混合物。照射后,通过使用碘化丙啶(PI)和荧光素二乙酸酯(FDA)的流式细胞术检测来评估膜通透性。通透-重封细胞的比例受光强度(约780毫瓦/平方厘米)、照射时间(约240秒)和/或颗粒浓度(约10⁹颗粒/毫升)变化的影响。光照射后通透性可持续长达20分钟。在单个颗粒的部位附近,细胞膜的通透性发生改变,这可能是由于局部温度变化所致。这些结果表明,这种激光诱导的通透策略构成了一种将外源物质递送至活细胞的新方法。