Department of Bioengineering, the University of Tokyo, Japan.
Electrophoresis. 2011 Sep;32(18):2496-501. doi: 10.1002/elps.201100129. Epub 2011 Aug 23.
In this paper, we present a novel electrofusion device that enables massive parallelism, using an electrically insulating sheet having a two-dimensional micro-orifice array. The sheet is sandwiched by a pair of micro-chambers with immersed electrodes, and each chamber is filled with the suspensions of the two types of cells to be fused. Dielectrophoresis, assisted by sedimentation, is used to position the cells in the upper chamber down onto the orifices, then the device is flipped over to position the cells on the other side, so that cell pairs making contact in the orifice are formed. When a pulse voltage is applied to the electrodes, most voltage drop occurs around the orifice and impressed on the cell membrane in the orifice. This makes possible the application of size-independent voltage to fuse two cells in contact at all orifices exclusively in 1:1 manner. In the experiment, cytoplasm of one of the cells is stained with a fluorescence dye, and the transfer of the fluorescence to the other cell is used as the indication of fusion events. The two-dimensional orifice arrangement at the pitch of 50 μm realizes simultaneous fusion of 6 × 10³ cells on a 4 mm diameter chip, and the fusion yield of 78-90% is achieved for various sizes and types of cells.
本文提出了一种新颖的电融合装置，该装置利用具有二维微孔阵列的电绝缘片实现大规模并行处理。该薄片被夹在一对带有浸入式电极的微腔之间，每个腔室中都充满了要融合的两种类型细胞的悬浮液。通过沉降辅助的介电泳将细胞定位在上腔室中的孔上，然后将装置翻转，使细胞位于另一侧，从而在孔中形成接触的细胞对。当在电极上施加脉冲电压时，大部分电压降发生在孔周围，并施加在孔中的细胞膜上。这使得在所有孔中以 1:1 的方式仅对接触的两个细胞施加独立于尺寸的电压进行融合成为可能。在实验中，用荧光染料染色其中一个细胞的细胞质，将荧光转移到另一个细胞作为融合事件的指示。在 50 μm 的节距处的二维孔排列实现了在 4mm 直径的芯片上同时融合 6×10³个细胞，并且对于各种大小和类型的细胞，融合产率达到了 78-90%。
