Application of dynamic diffractive optics for enhanced femtosecond laser based cell transfection
Application of dynamic diffractive optics for enhanced femtosecond laser based cell transfection
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DOI：
10.1002/jbio.201000052
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发表时间：
2010-10-01
影响因子：
2.8
通讯作者：
Dholakia, Kishan
中科院分区：
文献类型：
--
作者：
Antkowiak, Maciej;Torres-Mapa, Maria Leilani;Dholakia, Kishan
我们展示了动态衍射光学元件的优点，即空间光调制器（SLM）的控制和增强的光注入和光转染的哺乳动物细胞与飞秒光源。SLM提供对光束的横向和轴向定位的完全控制，精度达到亚微米。快速光束平移使得能够进行时间序列照射，这被示出为提高光注入效率并减轻在细胞膜上精确光束定位的问题。我们发现，在三个轴向位置的照射时，与单剂量相比，可光注入细胞的数量增加了一倍。所提出的系统还能够实现非靶向光栅扫描照射，其以每秒1个细胞的速率提供贴壁细胞的更高通量转染。此外，荧光成像用于展示细胞选择性两步基因治疗。[图]用表达Mito-DsRed的质粒选择性光转染的表达绿色荧光蛋白（GFP）的CHO-K1细胞。
We demonstrate the advantages of a dynamic diffractive optical element, namely a spatial light modulator (SLM) for the controlled and enhanced optoinjection and phototransfection of mammalian cells with a femtosecond light source. The SLM provides full control over the lateral and axial positioning of the beam with sub-micron precision. Fast beam translation enables time-sequenced irradiation, which is shown to enhance the optoinjection efficiency and alleviate the problem of exact beam positioning on the cell membrane. We show that irradiation in three axial positions doubles the number of viably optoinjected cells when compared with a single dose. The presented system also enables untargeted raster scan irradiation which provides a higher throughput transfection of adherent cells at the rate of 1 cell per second. Additionally, fluorescent imaging is used to demonstrate cell selective two-step gene therapy.[GRAPHICS]A Green Flourescent Protein (GFP) expressing CHO-K1 cell selectively phototransfected with a plasmid expressing Mito-DsRed.