Ionescu-Zanetti Cristian, Blatz Andrew, Khine Michelle
Fluxion Biosciences, San Francisco, CA 94158, USA.
Biomed Microdevices. 2008 Feb;10(1):113-6. doi: 10.1007/s10544-007-9115-x.
Single-cell electroporation, in which a focused electric field is applied to permeabilize an individual target cell using relatively low applied voltages, has demonstrated improved cell viability and transfection rates over conventional bulk electroporation set-ups. Here, we introduce a new strategy, in conjunction with single-cell electroporation, to enhance exogenous transport efficiency: electrophoresis delivery of compounds subsequent to electroporation. Electrophoresis is used to assist loading of otherwise impermeable exogenous anionic fluorescent molecules Calcein (Invitrogen, MW = 622) and Oregon Green Dextran (OGD, Invitrogen, MW = 70,000). For the larger dextran molecules, we demonstrate a protocol of first pre-concentrating at the cell-microfluidic channel interface. Then, the electric field is used to drive these molecules into the cell post-electroporation using 50-200 mV. We demonstrate delivery rate enhancements of more than an order of magnitude using electrophoresis compared to diffusion alone subsequent to electroporation.
单细胞电穿孔是指施加聚焦电场，利用相对较低的施加电压使单个靶细胞通透化，与传统的批量电穿孔装置相比，已证明其细胞活力和转染率有所提高。在此，我们介绍一种与单细胞电穿孔相结合的新策略，以提高外源转运效率：电穿孔后通过电泳递送化合物。电泳用于辅助加载原本不可渗透的外源阴离子荧光分子钙黄绿素（赛默飞世尔科技，分子量 = 622）和俄勒冈绿葡聚糖（OGD，赛默飞世尔科技，分子量 = 70,000）。对于较大的葡聚糖分子，我们展示了一种先在细胞 - 微流体通道界面进行预浓缩的方案。然后，使用50 - 200 mV的电场在电穿孔后将这些分子驱动到细胞中。与电穿孔后仅通过扩散相比，我们证明使用电泳可使递送速率提高一个数量级以上。
