Kim Kwang Bok, Chun Honggu, Kim Hee Chan, Chung Taek Dong
Graduate School, Seoul National University, Seoul, Korea.
Electrophoresis. 2009 May;30(9):1464-9. doi: 10.1002/elps.200800448.
This paper reports on a novel microfluidic chip with polyelectrolytic gel electrodes (PGEs) used to rapidly count the number of red blood cells (RBCs) in diluted whole blood. The proposed microdevice is based on the principle that the impedance across a microchannel between two PGEs varies sensitively as RBCs pass through it. The number and amplitude of impedance peaks provide the information about the number and size of RBCs, respectively. This system features a low-voltage dc detection method and non-contact condition between cells and metal electrodes. Major advantages include stable detection under varying cellular flow rate and position in the microchannel, little chance of cell damage due to high electric field gradient and no surface fouling of the metal electrodes. The performance of this PGEs-based system was evaluated in three steps. First, in order to observe the size-only dependence of the impedance signal, three different sizes of fluorescent microbeads (7.2, 10.0, and 15.0 mum; Bangs laboratories, USA) were used in the experiment. Second, the cell counting performance was evaluated by using 7.2 mum fluorescent microbeads, similar in size to RBCs, in various concentrations and comparing the results with an animal hematoanalyzer (MS 9-5; Melet schloesing laboratories, France). Finally, in human blood sample tests, intravenously collected whole blood was just diluted in a PBS without centrifuge or other pretreatments. The PGE-based system produced almost identical number of RBCs in over 800-fold diluted samples to the results from a commercialized human hematoanalyzer (HST-N402XE; Sysmex, Japan).
本文报道了一种带有聚电解质凝胶电极(PGEs)的新型微流控芯片,用于快速计数稀释全血中的红细胞(RBCs)数量。所提出的微型设备基于这样的原理:当红细胞通过两个PGEs之间的微通道时,微通道两端的阻抗会敏感变化。阻抗峰值的数量和幅度分别提供了有关红细胞数量和大小的信息。该系统具有低压直流检测方法以及细胞与金属电极之间的非接触条件。主要优点包括在微通道中细胞流速和位置变化时检测稳定、由于高电场梯度导致细胞损伤的可能性小以及金属电极无表面污染。基于PGEs的该系统性能分三步进行评估。首先,为了观察阻抗信号仅与尺寸的相关性,实验中使用了三种不同尺寸的荧光微珠(7.2、10.0和15.0μm;美国Bangs实验室)。其次,通过使用尺寸与红细胞相似的7.2μm荧光微珠,在不同浓度下评估细胞计数性能,并将结果与动物血液分析仪(MS 9 - 5;法国Melet schloesing实验室)进行比较。最后,在人体血液样本测试中,静脉采集的全血仅在磷酸盐缓冲盐溶液(PBS)中稀释,无需离心或其他预处理。基于PGEs的系统在稀释超过800倍的样本中产生的红细胞数量与商业化人体血液分析仪(HST - N402XE;日本Sysmex)的结果几乎相同。