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渗透应激下的细胞内钾决定了兆赫兹范围内小鼠骨髓瘤细胞的介电泳交叉频率。

Intracellular potassium under osmotic stress determines the dielectrophoresis cross-over frequency of murine myeloma cells in the MHz range.

作者信息

Chung Colin, Pethig Ronald, Smith Stewart, Waterfall Martin

机构信息

School of Engineering, Institute for Integrated Micro & Nanosystems, Edinburgh, UK.

Institute for Bioengineering, University of Edinburgh, Edinburgh, UK.

出版信息

Electrophoresis. 2018 Apr;39(7):989-997. doi: 10.1002/elps.201700433. Epub 2018 Jan 15.

DOI:10.1002/elps.201700433
PMID:29274244
Abstract

Dielectrophoresis (DEP) has been widely studied for its potential as a biomarker-free method of sorting and characterizing cells based upon their dielectric properties. Most studies have employed voltage signals from ∼1 kHz to no higher than ∼30 MHz. Within this range a transition from negative to positive DEP can be observed at the cross-over frequency f . The value of f is determined by the conductivity of the suspending medium, as well as the size and shape of the cell and the dielectric properties (capacitance, conductivity) of its plasma membrane. In this work DEP measurements were performed up to 400 MHz, where the transition from positive to negative DEP can be observed at a higher cross-over frequency f . SP2/O murine myeloma cells were suspended in buffer media of different osmolarities and measurements taken of cell volume, f and f . Potassium-binding benzofuran isophthalate (PBFI), a potassium-sensitive fluorophore, and flow cytometry was employed to monitor relative changes in intracellular potassium concentration. In agreement with theory, it was found that f is independent of the cell parameters that control f and is predominantly determined by intracellular conductivity. In particular, the value of f is highly correlated to that of the intracellular potassium concentration.

摘要

介电泳(DEP)因其作为一种基于细胞介电特性对细胞进行分选和表征的无生物标志物方法的潜力而受到广泛研究。大多数研究采用的电压信号频率范围约为1kHz至不高于约30MHz。在此范围内,在交叉频率f处可观察到从负介电泳到正介电泳的转变。f的值由悬浮介质的电导率以及细胞的大小和形状及其质膜的介电特性(电容、电导率)决定。在这项工作中,介电泳测量进行到了400MHz,在该频率下,可在更高的交叉频率f处观察到从正介电泳到负介电泳的转变。将SP2/O小鼠骨髓瘤细胞悬浮在不同渗透压的缓冲介质中,并测量细胞体积、f和f。使用钾结合苯并呋喃间苯二甲酸酯(PBFI),一种钾敏感荧光团,以及流式细胞术来监测细胞内钾浓度的相对变化。与理论一致,发现f与控制f的细胞参数无关,并且主要由细胞内电导率决定。特别是,f的值与细胞内钾浓度的值高度相关。

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