Single cell transfection using plasmid decorated AFM probes
Single cell transfection using plasmid decorated AFM probes
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DOI：
10.1016/j.bbrc.2007.01.190
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发表时间：
2007-04-13
影响因子：
3.1
通讯作者：
Grandbois, Michel
中科院分区：
文献类型：
--
作者：
Cuerrier, Charles M.;Lebel, Rejean;Grandbois, Michel
使用市售的原子力显微镜尖端分别转染真核细胞，该尖端装饰有编码荧光蛋白 EGFP 的质粒 DNA。在典型的转染尝试中，尖端被强行掺入细胞中，从而允许遗传物质通过细胞膜转移。当监测细胞变形作为施加力的函数时，可以很容易地检测到与尖端穿过细胞膜的通道相对应的尖锐不连续性。为了使转染成功，尖端必须可逆地穿透膜而不会对细胞造成干扰或损坏。通过落射荧光显微镜证实转染成功率（30%）、细胞存活和生长。该技术为转染工具箱提供了一种替代工具，允许以最小的干扰转染特定的单个细胞。 (c) 2007 Elsevier Inc. 保留所有权利。
Eukaryotic cells were individually transfected using commercially available atomic force microscope tips decorated with plasmidic DNA encoding for the fluorescent protein EGFP. In a typical transfection attempt, the tip is forcibly incorporated into the cell thus allowing for the transfer of the genetic material through the cell membrane. A sharp discontinuity, corresponding to the passage of the tip through the cell membrane, can be easily detected when monitoring the cellular deformation as a function of the applied force. In order for the transfection to be successful, the tip must reversibly penetrates the membrane without causing disturbance or damage to the cell. Transfection success rate (30%), cell survival, and growth are confirmed by epifluorescence microscopy. This technique provides an alternative tool to the transfection toolbox, allowing the transfection of specific individual cells with minimal disturbance. (c) 2007 Elsevier Inc. All rights reserved.