Sabuncu Ahmet C, Asmar Anthony J, Stacey Michael W, Beskok Ali
Department of Mechanical Engineering, Southern Methodist University, Dallas, VA, USA.
Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, USA.
Electrophoresis. 2015 Jul;36(13):1499-506. doi: 10.1002/elps.201500119. Epub 2015 Jun 12.
Electromanipulation of cells as a label-free cell manipulation and characterization tool has gained particular interest recently. However, the applicability of electromanipulation, particularly dielectrophoresis (DEP), to biological cells is limited to cells suspended in buffers containing lower amounts of salts relative to the physiological buffers. One might question the use of low conductivity buffers (LCBs) for DEP separation, as cells are stressed in buffers lacking physiological levels of salt. In LCB, cells leak ions and undergo volume regulation. Therefore, cells exhibit time-dependent DEP response in LCB. In this work, cellular changes in LCB are assessed by dielectric spectroscopy, cell viability assay, and gene expression of chondrocytes and Jurkats. Results indicate leakage of ions from cells, increases in cytoplasmic conductivity, membrane capacitance, and conductance. Separability factor, which defines optimum conditions for DEP cell separation, for the two cell types is calculated using the cellular dielectric data. Optimum DEP separation conditions change as cellular dielectric properties evolve in LCB. Genetic analyses indicate no changes in expression of ionic channel proteins for chondrocytes suspended in LCB. Retaining cellular viability might be important during dielectrophoretic separation, especially when cells are to be biologically tested at a downstream microfluidic component.
作为一种无标记细胞操作和表征工具,细胞电操纵最近引起了特别关注。然而,电操纵,特别是介电电泳(DEP)对生物细胞的适用性仅限于悬浮在相对于生理缓冲液含盐量较低的缓冲液中的细胞。有人可能会质疑使用低电导率缓冲液(LCB)进行DEP分离,因为细胞在缺乏生理盐水平的缓冲液中会受到应激。在LCB中,细胞会泄漏离子并进行体积调节。因此,细胞在LCB中表现出随时间变化的DEP响应。在这项工作中,通过介电谱、细胞活力测定以及软骨细胞和 Jurkat 细胞的基因表达来评估LCB中的细胞变化。结果表明细胞离子泄漏、细胞质电导率、膜电容和电导增加。使用细胞介电数据计算了两种细胞类型的可分离性因子,该因子定义了DEP细胞分离的最佳条件。随着细胞介电特性在LCB中演变,最佳DEP分离条件也会改变。基因分析表明,悬浮在LCB中的软骨细胞离子通道蛋白表达没有变化。在介电电泳分离过程中保持细胞活力可能很重要,尤其是当要在下游微流体组件中对细胞进行生物学测试时。