Yamane Daisuke, Wu Yi-Chien, Wu Ting-Hsiang, Toshiyoshi Hiroshi, Teitell Michael A, Chiou Pei-Yu
1Precision and Intelligence Laboratory, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.
J Lab Autom. 2014 Feb;19(1):50-9. doi: 10.1177/2211068213494390. Epub 2013 Jun 24.
To transfer large cargo into mammalian cells, we recently provided a new approach called a photothermal nanoblade. Micron-sized membrane pores generated by the nanoblade are surprisingly well repaired with little cell death, suggesting rapid membrane-resealing dynamics. Here, we report the resealing time of photothermal porated mammalian cell plasma membranes using an electrical impedance sensor. Cell membrane pores were generated by high-speed cavitation bubbles induced by laser pulsing of metallic microdisks on a pair of transparent indium tin oxide electrodes. Electrical responses from the sensor electrodes were obtained with a signal voltage of 500 mV and a frequency at 500 kHz. Real-time impedance measurements show that membrane resealing and impedance recovery take a surprisingly long 1 to 2 min after laser pulsing. A nonrecovering impedance shift is also detected for cells after high-energy laser pulsing. This impedance response is also confirmed by a separate experiment in which thin-film gold electrodes are used to trigger cavitation bubbles for opening transient membrane pores on cells cultured on electrodes. Overall, our study platform provides new insight for micron-sized membrane defect repair dynamics to maintain cell viability.
为了将大分子货物转运到哺乳动物细胞中,我们最近提出了一种名为光热纳米刀片的新方法。纳米刀片产生的微米级膜孔能以极少的细胞死亡得到惊人良好的修复,这表明存在快速的膜重封动力学。在此,我们使用电阻抗传感器报告了光热穿孔的哺乳动物细胞质膜的重封时间。细胞膜孔是由一对透明氧化铟锡电极上的金属微盘激光脉冲诱导产生的高速空化气泡形成的。传感器电极的电响应通过500 mV的信号电压和500 kHz的频率获得。实时阻抗测量表明,激光脉冲后膜重封和阻抗恢复需要长达1至2分钟的惊人时间。在高能激光脉冲后,细胞还检测到不可逆的阻抗变化。在另一个实验中也证实了这种阻抗响应,该实验使用薄膜金电极触发空化气泡,以在电极上培养的细胞上打开瞬时膜孔。总体而言,我们的研究平台为微米级膜缺陷修复动力学以维持细胞活力提供了新的见解。