Departamento de Biotecnología e Ingeniería de Alimentos y Centro de Biotecnología, Tecnológico de Monterrey, Monterrey, Nuevo León, México.
Electrophoresis. 2009 Dec;30(24):4195-205. doi: 10.1002/elps.200900355.
Electrokinetic techniques offer a great potential for biological particle manipulation. Among these, dielectrophoresis (DEP) has been successfully utilized for the concentration of bioparticles. Traditionally, DEP is performed employing microelectrodes, an approach with attractive characteristics but expensive due to microelectrode fabrication costs. An alternative is insulator-based DEP, a method where non-uniform electric fields are created with arrays of insulating structures. This study presents the concentration of linear DNA particles (pET28b) employing a microchannel, with an array of cylindrical insulating structures and direct current electric fields. Results showed manipulation of DNA particles with a combination of electroosmotic, electrophoretic, and dielectrophoretic forces. Employing suspending media with conductivity of 104 muS/cm and pH of 11.15, under applied fields between 500 and 1500 V/cm, DNA particles were observed to be immobilized due to negative dielectrophoretic trapping. The observation of DNA aggregates that occurred at higher applied fields, and dispersed once the field was removed is also included. Finally, concentration factors varying from 8 to 24 times the feed concentration were measured at 2000 V/cm after concentration time-periods of 20-40 s. The results presented here demonstrate the potential of insulator-based DEP for DNA concentration, and open the possibility for fast DNA manipulation for laboratory and large-scale applications.
电泳技术为生物粒子操纵提供了巨大的潜力。其中,介电泳(DEP)已成功用于生物粒子的浓缩。传统上,DEP 是通过微电极来实现的,这种方法具有吸引力的特点,但由于微电极制造费用昂贵。另一种方法是基于绝缘体的 DEP,它是一种使用绝缘结构阵列产生非均匀电场的方法。本研究采用带有圆柱形绝缘结构阵列和直流电场的微通道,对线性 DNA 颗粒(pET28b)进行了浓缩。结果表明,可以通过电动、电泳和介电泳力来操纵 DNA 颗粒。在 104 μS/cm 的电导率和 11.15 的 pH 值的悬浮介质中,在 500 到 1500 V/cm 的应用场下,DNA 颗粒由于负介电泳捕获而被固定。还观察到在较高的应用场下发生 DNA 聚集,一旦去除电场就会分散。最后,在 2000 V/cm 的应用场下,在 20-40 s 的浓缩时间后,测量到的浓缩因子为进料浓度的 8 到 24 倍。这里呈现的结果表明,基于绝缘体的 DEP 具有用于 DNA 浓缩的潜力,并为实验室和大规模应用中的快速 DNA 操作开辟了可能性。