A single cell electroporation chip
A single cell electroporation chip
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DOI：
10.1039/b408352k
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发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Lee, LP
中科院分区：
文献类型：
--
作者：
Khine, M;Lau, A;Lee, LP
增加细胞膜的渗透性可以通过单细胞电穿孔来实现。因此，不能渗透质膜的极性物质（如染料、药物、DNA、蛋白质、肽和氨基酸）可以被引入细胞。我们开发了一种聚合物芯片，可以选择性地固定和局部电穿孔单细胞。这种易于使用的芯片可以聚焦电场，无需操作电极或玻璃移液器。此外，该装置允许平行的单细胞电穿孔。我们通过使用低施加电压（<1V）电穿孔HeLa细胞来证明我们的装置设计的有效性。我们发现HeLa细胞电穿孔所需的平均跨膜电位为0.51 +/- 0.13 V。膜渗透通过测量对应于细胞电阻下降的电流中的特征“跳跃”进行电评估，并且通过记录细胞质染料钙黄绿素AM的逃逸或台盼蓝染色的进入进行显微镜评估。
Increasing the cell membrane's permeability can be accomplished via single cell electroporation. Polar substances that cannot otherwise permeate the plasma membrane ( such as dyes, drugs, DNA, proteins, peptides, and amino acids) can thus be introduced into the cell. We developed a polymeric chip that can selectively immobilize and locally electroporate single cells. This easy-to-use chip focuses the electric field, eliminating the need to manipulate electrodes or glass pipettes. Moreover, this device allows parallel single cell electroporation. We demonstrate the effectiveness of our device design by electroporating HeLa cells using low applied voltages (< 1 V). We found the average transmembrane potential required for electroporation of HeLa cells to be 0.51 +/- 0.13 V. Membrane permeation is assessed electrically by measuring characteristic 'jumps' in current that correspond to drops in cell resistance, and microscopically by recording either the escape of cytoplasmic dye Calcein AM or the entrance of Trypan blue stain.