Olofsson Jessica, Levin Mikael, Strömberg Anette, Weber Stephen G, Ryttsén Frida, Orwar Owe
Department of Physical Chemistry, Chalmers University of Technology, Kemivägen 10, SE-412 96 Gothenburg, Sweden.
Anal Chem. 2007 Jun 15;79(12):4410-8. doi: 10.1021/ac062140i. Epub 2007 May 19.
We present a computer-controlled scanning electroporation method. Adherent cells are electroporated using an electrolyte-filled capillary in contact with an electrode. The capillary can be scanned over a cell culture and locally deliver both an electric field and an electroporation agent to the target area without affecting surrounding cells. The instantaneous size of the targeted area is determined by the dimensions of the capillary. The size and shape of the total electroporated area are defined by these dimensions in combination with the scanning pattern. For example, striped and serpentine patterns of electroporated cells in confluent cultures can be formed. As it is easy to switch between different electroporation agents, the method is suitable for design of cell cultures with complex composition. Finite element method simulations were used to study the spatial distributions of the electric field and the concentration of an electroporation agent, as well as the fluid dynamics related to scanning and flow of electroporation agent from the capillary. The method was validated for transfection by introduction of a 9-base-pair-long randomized oligonucleotide into PC12 cells and a pmaxGFP plasmid coding for green fluorescent protein into CHO and WSS cells.
我们提出了一种计算机控制的扫描电穿孔方法。使用与电极接触的充满电解质的毛细管对贴壁细胞进行电穿孔。毛细管可以在细胞培养物上扫描，并将电场和电穿孔剂局部递送至目标区域，而不影响周围细胞。目标区域的瞬时大小由毛细管的尺寸决定。总电穿孔区域的大小和形状由这些尺寸与扫描模式共同定义。例如，在汇合培养物中可以形成电穿孔细胞的条纹状和蜿蜒状图案。由于易于在不同的电穿孔剂之间切换，该方法适用于设计具有复杂组成的细胞培养物。使用有限元方法模拟来研究电场和电穿孔剂浓度的空间分布，以及与扫描和电穿孔剂从毛细管流动相关的流体动力学。通过将9个碱基对长的随机寡核苷酸引入PC12细胞，并将编码绿色荧光蛋白的pmaxGFP质粒引入CHO和WSS细胞，验证了该方法用于转染的有效性。
