Transient transfection of mammalian cells using a violet diode laser.
Transient transfection of mammalian cells using a violet diode laser.
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DOI：
10.1117/1.3430730
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发表时间：
2010-07
影响因子：
3.5
通讯作者：
M. L. Torres-Mapa;Liselotte Angus;M. Ploschner;K. Dholakia;F. Gunn-Moore
中科院分区：
文献类型：
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作者：
M. L. Torres-Mapa;Liselotte Angus;M. Ploschner;K. Dholakia;F. Gunn-Moore
我们证明了第一次使用的紫色二极管激光瞬时哺乳动物细胞转染。与先前的研究相比，这些研究显示了在几周内产生稳定的细胞系，我们开发了一种方法来瞬时转染细胞，效率高达约40%。在编码线粒体靶向红色荧光蛋白的质粒DNA存在下，将中国仓鼠卵巢（CHO-K1）和人胚肾（HEK 293）细胞暴露于紧密聚焦的405 nm激光。我们报告转染效率作为我们的系统的激光功率和曝光时间的函数。我们还表明，第一次，连续波激光源可以成功地应用于选择性基因沉默实验，使用小干扰RNA。这项工作是朝着廉价和便携式光转染系统迈出的重要一步。
We demonstrate the first use of the violet diode laser for transient mammalian cell transfection. In contrast to previous studies, which showed the generation of stable cell lines over a few weeks, we develop a methodology to transiently transfect cells with an efficiency of up to approximately 40%. Chinese hamster ovary (CHO-K1) and human embryonic kidney (HEK293) cells are exposed to a tightly focused 405-nm laser in the presence of plasmid DNA encoding for a mitochondrial targeted red fluorescent protein. We report transfection efficiencies as a function of laser power and exposure time for our system. We also show, for the first time, that a continuous wave laser source can be successfully applied to selective gene silencing experiments using small interfering RNA. This work is a major step towards an inexpensive and portable phototransfection system.