Microsystems Laboratory, Ecole Polytechnique Federale de Lausanne (EPFL), BM 3.124 Station 17, Lausanne, Vaud 1015, Switzerland.
Biomicrofluidics. 2010 Jun 29;4(2):022807. doi: 10.1063/1.3430542.
There is great interest in highly sensitive separation methods capable of quickly isolating a particular cell type within a single manipulation step prior to their analysis. We present a cell sorting device based on the opposition of dielectrophoretic forces that discriminates between cell types according to their dielectric properties, such as the membrane permittivity and the cytoplasm conductivity. The forces are generated by an array of electrodes located in both sidewalls of a main flow channel. Cells with different dielectric responses perceive different force magnitudes and are, therefore, continuously focused to different equilibrium positions in the flow channel, thus avoiding the need of a specific cell labeling as discriminating factor. We relate the cells' dielectric response to their output position in the downstream channel. Using this microfluidic platform that integrates a method of continuous-flow cell separation based on multiple frequency dielectrophoresis, we succeeded in sorting viable from nonviable yeast with nearly 100% purity. The method also allowed to increase the infection rate of a cell culture up to 50% of parasitemia percentage, which facilitates the study of the parasite cycle. Finally, we prove the versatility of our device by synchronizing a yeast cell culture at a particular phase of the cell cycle avoiding the use of metabolic agents interfering with the cells' physiology.
人们对能够在单个操作步骤内快速分离特定细胞类型的高度敏感分离方法非常感兴趣,然后再对其进行分析。我们提出了一种基于介电泳力对抗的细胞分选装置,该装置根据细胞的介电特性(例如膜介电常数和细胞质电导率)来区分细胞类型。力是由位于主流动通道两侧壁上的电极阵列产生的。具有不同介电响应的细胞感知到不同的力大小,因此,它们会在流动通道中连续聚焦到不同的平衡位置,从而避免了将特定的细胞标记作为区分因素的需要。我们将细胞的介电响应与其在下游通道中的输出位置联系起来。使用这种集成了基于多频介电泳的连续流动细胞分离方法的微流控平台,我们成功地以近 100%的纯度从非存活酵母中分离出存活酵母。该方法还可以将细胞培养物的感染率提高到寄生虫百分比的 50%,这有助于寄生虫周期的研究。最后,我们通过在细胞周期的特定阶段同步酵母细胞培养物来证明我们的设备的多功能性,从而避免使用干扰细胞生理的代谢试剂。