Nanofountain probe electroporation (NFP-E) of single cells.
Nanofountain probe electroporation (NFP-E) of single cells.
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DOI：
10.1021/nl400423c
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发表时间：
2013-06-12
期刊：
影响因子：
10.8
通讯作者：
Espinosa HD
中科院分区：
文献类型：
--
作者：
Kang W;Yavari F;Minary-Jolandan M;Giraldo-Vela JP;Safi A;McNaughton RL;Parpoil V;Espinosa HD
精确地将分子递送到单细胞中的能力对于生物技术研究人员来说是非常感兴趣的，以推进在治疗学、诊断学和药物递送中的应用，从而实现个性化医疗的前景。在过去的十年中，大量电穿孔技术用于细胞转染的使用显著增加，但该技术是非特异性的并且需要高电压，导致可变的效率和低的细胞活力。我们已经开发了一种使用纳米峰探针（NFP）技术进行电穿孔的新工具，该技术可以以比批量电穿孔或显微注射更高效且对细胞更温和的方式将分子递送到细胞中。在这里，我们证明了NFP电穿孔（NFP-E）的单个HeLa细胞内的人口，他们与荧光标记的葡聚糖和成像的细胞，以评估转染效率和细胞活力。我们的NFP-E的机制的理论分析表明，电压的应用程序创建一个本地化的电场之间的NFP悬臂尖端和细胞膜的区域与尖端接触。因此，NFP-E可以将分子递送到靶细胞，而对细胞的电势影响最小。我们在HeLa细胞上的实验证实，NFP-E提供单细胞选择性、高转染效率（>95%）、定性剂量控制和转染细胞的非常高的存活率（92%）。
The ability to precisely deliver molecules into single cells is of great interest to biotechnology researchers for advancing applications in therapeutics, diagnostics, and drug delivery toward the promise of personalized medicine. The use of bulk electroporation techniques for cell transfection has increased significantly in the last decade, but the technique is nonspecific and requires high voltage, resulting in variable efficiency and low cell viability. We have developed a new tool for electroporation using nanofountain probe (NFP) technology, which can deliver molecules into cells in a manner that is highly efficient and gentler to cells than bulk electroporation or microinjection. Here we demonstrate NFP electroporation (NFP-E) of single HeLa cells within a population by transfecting them with fluorescently labeled dextran and imaging the cells to evaluate the transfection efficiency and cell viability. Our theoretical analysis of the mechanism of NFP-E reveals that application of the voltage creates a localized electric field between the NFP cantilever tip and the region of the cell membrane in contact with the tip. Therefore, NFP-E can deliver molecules to a target cell with minimal effect of the electric potential on the cell. Our experiments on HeLa cells confirm that NFP-E offers single cell selectivity, high transfection efficiency (>95%), qualitative dosage control, and very high viability (92%) of transfected cells.
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影响因子：
29.4
作者：
Kim, Keun-Ho;Sanedrin, Raymond G.;Espinosa, Horacio D.
通讯作者：
Espinosa, Horacio D.
DOI：
10.1073/pnas.0909250107
发表时间：
2010-03-30
影响因子：
11.1
作者：
Almquist, Benjamin D.;Melosh, Nicholas A.
通讯作者：
Melosh, Nicholas A.
影响因子：
6.1
作者：
Khine, M;Lau, A;Lee, LP
通讯作者：
Lee, LP
影响因子：
7.4
作者：
Fei, Zhengzheng;Hu, Xin;Lee, L. James
通讯作者：
Lee, L. James
影响因子：
3.5
作者：
GRAESSMANN, A;GRAESSMANN, M;MUELLER, N
通讯作者：
MUELLER, N