He Hao, Kong Siu-Kai, Lee Rebecca Kit-Ying, Suen Yick-Keung, Chan Kam Tai
Department of Electronic Engineering, The Chinese University of Hong Kong, Shatin, Hong Kong.
Opt Lett. 2008 Dec 15;33(24):2961-3. doi: 10.1364/ol.33.002961.
We report the transfection of human hepatocarcinoma (HepG2) cells by femtosecond (fs) laser pulses at 1554 nm. It was found that HepG2 cells could be perforated transiently to admit propidium iodide by that laser. This photoporation was safe, as the membrane resealed itself within a short time and no mitochondrial depolarization was detected. The cells were next photoporated in the presence of DNA plasmids for the expression of green fluorescence protein, and about 80% of the exposed cells showed green fluorescence 24 h later. Thus it could be concluded that it is safe and efficient to use fs laser at 1554 nm to transfect foreign molecules into cells under a standard microscope.
我们报道了在1554nm波长下用飞秒(fs)激光脉冲转染人肝癌(HepG2)细胞。结果发现,该激光可使HepG2细胞瞬时穿孔以摄取碘化丙啶。这种光穿孔是安全的,因为细胞膜在短时间内自行重新封闭,且未检测到线粒体去极化。接下来,在存在用于表达绿色荧光蛋白的DNA质粒的情况下对细胞进行光穿孔,约80%的暴露细胞在24小时后发出绿色荧光。因此可以得出结论,在标准显微镜下使用1554nm的飞秒激光将外源分子转染到细胞中是安全且高效的。