High-Throughput Continuous Flow Femtosecond Laser-Assisted Cell Optoporation and Transfection
High-Throughput Continuous Flow Femtosecond Laser-Assisted Cell Optoporation and Transfection
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DOI：
10.1002/jemt.22423
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发表时间：
2014-12-01
影响因子：
2.5
通讯作者：
Koenig, Karsten
中科院分区：
文献类型：
--
作者：
Breunig, Hans Georg;Uchugonova, Aisada;Koenig, Karsten
我们提出了一种基于飞秒激光的纳米处理系统，用于瞬态光学细胞膜穿孔，以非常高的通量将外来分子引入细胞内部。在该装置中，细胞通过微流管进行空间限制，同时被激光辐射靶向。波束整形沿着一条线产生焦点几何形状，这条线被扫描穿过微流单元以增加可到达的单元的数量。通过细胞转染，即使细胞-光相互作用时间在毫秒范围内，也可以间接观察到成功的细胞膜穿孔。以中国仓鼠卵巢细胞为实验对象，考察了该体系的细胞活力、外源分子的摄取和细胞转染效率。与之前处理数百万个细胞的非流动方法相比，细胞的连续流动使细胞吞吐量大大增加，尽管效率有限。该装置开启了实现完全自动化的高通量激光辅助细胞穿孔系统的可能性，该系统可以集成在芯片上的实验室设备中。Microsc。科学通报，2014(7):974-979。(c) 2014 Wiley期刊公司
We present a femtosecond-laser based nanoprocessing system for transient optical cell membrane poration to allow the introduction of foreign molecules into the interior of a cell with very high throughput. In the setup, cells flow through a micro-flow tube for spatial confinement and are simultaneously targeted by fs laser radiation. Beam-shaping generates a focal geometry along a line which is scanned across the micro-flow cell to increase the number of reachable cells. Successful cell membrane poration was observed indirectly by cell transfection even with cell-light interaction times in the millisecond range. The system was characterized by experiments with Chinese hamster ovary cells regarding cell viability, the uptake of extrinsic molecules and cell transfection efficiency. The continuous flow of cells enables a tremendous increase of cell throughput compared to previous nonflow approaches by treating millions of cells, although with only limited efficiency. The setup opens the possibility to realize a completely automated high-throughput laser-assisted cell-poration system which could be integrated in lab-on-a-chip devices. Microsc. Res. Tech. 77:974-979, 2014. (c) 2014 Wiley Periodicals, Inc.