Department of Electrical and Computer Engineering, University of Manitoba, Winnipeg, Manitoba R3T 5V6, Canada.
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada.
Biomicrofluidics. 2013 Mar 1;7(2):24101. doi: 10.1063/1.4793223. eCollection 2013.
The instrument described here is an all-electronic dielectrophoresis (DEP) cytometer sensitive to changes in polarizability of single cells. The important novel feature of this work is the differential electrode array that allows independent detection and actuation of single cells within a short section ([Formula: see text]) of the microfluidic channel. DEP actuation modifies the altitude of the cells flowing between two altitude detection sites in proportion to cell polarizability; changes in altitude smaller than 0.25 μm can be detected electronically. Analysis of individual experimental signatures allows us to make a simple connection between the Clausius-Mossotti factor (CMF) and the amount of vertical cell deflection during actuation. This results in an all-electronic, label-free differential detector that monitors changes in physiological properties of the living cells and can be fully automated and miniaturized in order to be used in various online and offline probes and point-of-care medical applications. High sensitivity of the DEP cytometer facilitates observations of delicate changes in cell polarization that occur at the onset of apoptosis. We illustrate the application of this concept on a population of Chinese hamster ovary (CHO) cells that were followed in their rapid transition from a healthy viable to an early apoptotic state. DEP cytometer viability estimates closely match an Annexin V assay (an early apoptosis marker) on the same population of cells.
这里描述的仪器是一种对单个细胞的极化率变化敏感的全电子介电泳(DEP)细胞仪。这项工作的一个重要新颖特点是差分电极阵列,它允许在微流道的短段([公式:见正文])内独立检测和驱动单个细胞。DEP 驱动会使在两个高度检测点之间流动的细胞的高度按比例改变,细胞高度的变化小于 0.25 μm 时可以通过电子方式检测到。对单个实验特征的分析使我们能够在 Clausius-Mossotti 因子(CMF)和驱动过程中细胞垂直偏转的量之间建立简单的联系。这导致了一种全电子、无标记的差分检测器,可监测活细胞生理特性的变化,并可完全自动化和微型化,以便用于各种在线和离线探头和即时医疗应用。DEP 细胞仪的高灵敏度促进了对细胞极化在细胞凋亡开始时发生的细微变化的观察。我们以一组中国仓鼠卵巢(CHO)细胞为例说明了这一概念的应用,这些细胞迅速从健康存活状态过渡到早期凋亡状态。DEP 细胞仪的存活率估计与同一细胞群上的 Annexin V 测定(早期凋亡标志物)非常吻合。