Xi'an Jiaotong University, School of Life Science and Technology, Institute of Biomedical Analytical Technology and Instrumentation, Xianning xi Road 28, Xi'an, 710049, China.
J Biomed Opt. 2009 Sep-Oct;14(5):054034. doi: 10.1117/1.3253320.
Light-absorbing nanoparticles that are heated by short laser pulses can transiently increase membrane permeability. We evaluate the membrane permeability by flow cytometry assaying of propidium iodide and fluorescein isothiocyanate dextran (FITC-D) using different laser sources. The dependence of the transfection efficiency on laser parameters such as pulse duration, irradiant exposure, and irradiation mode is investigated. For nano- and also picosecond irradiation, we show a parameter range where a reliable membrane permeabilization is achieved for 10-kDa FITC-D. Fluorescent labeled antibodies are able to penetrate living cells that are permeabilized using these parameters. More than 50% of the cells are stained positive for a 150-kDa IgG antibody. These results suggest that the laser-induced permeabilization approach constitutes a promising tool for targeted delivery of larger exogenous molecules into living cells.
短激光脉冲加热的光吸收纳米颗粒可以瞬时增加细胞膜的通透性。我们通过使用不同的激光源，用流式细胞术检测碘化丙啶和异硫氰酸荧光素右旋糖（FITC-D）来评估膜通透性。研究了转染效率对激光参数（如脉冲持续时间、辐照暴露和辐照模式）的依赖性。对于纳秒和皮秒辐射，我们展示了一个参数范围，在该范围内可以实现 10-kDa FITC-D 的可靠膜通透性。使用这些参数，荧光标记的抗体能够穿透通透性的活细胞。超过 50%的细胞对 150-kDa IgG 抗体呈阳性染色。这些结果表明，激光诱导的通透性方法为将较大的外源分子靶向递送至活细胞提供了一种很有前途的工具。
