Kim Minyoung, Rhee Hyunjoon, Kang Ji Yoon, Kim Tae Song, Kwak Rhokyun
Center for BioMicrosystems, Korea Institute of Science and Technology; Department of Mechanical Engineering, Seoul National University.
Center for BioMicrosystems, Korea Institute of Science and Technology; Department of Industrial Engineering, University of Illinois Urbana-Champaign.
J Vis Exp. 2017 Feb 23(120):55313. doi: 10.3791/55313.
The ion concentration polarization (ICP) phenomenon is one of the most prevailing methods to preconcentrate low-abundance biological samples. The ICP induces a noninvasive region for charged biomolecules (i.e., the ion depletion zone), and targets can be preconcentrated on this region boundary. Despite the high preconcentration performances with ICP, it is difficult to find the operating conditions of non-propagating ion depletion zones. To overcome this narrow operating window, we recently developed a new platform for spatiotemporally fixed preconcentration. Unlike preceding methods that only use ion depletion, this platform also uses the opposite polarity of the ICP (i.e., ion enrichment) to stop the propagation of the ion depletion zone. By confronting the enrichment zone with the depletion zone, the two zones merge together and stop. In this paper, we describe a detailed experimental protocol to build this spatiotemporally defined ICP platform and characterize the preconcentration dynamics of the new platform by comparing them with those of the conventional device. Qualitative ion concentration profiles and current-time responses successfully capture the different dynamics between the merged ICP and the stand-alone ICP. In contrast to the conventional one that can fix the preconcentration location at only ~5 V, the new platform can produce a target-condensed plug at a specific location in the broad ranges of operating conditions: voltage (0.5-100 V), ionic strength (1-100 mM), and pH (3.7-10.3).
离子浓度极化(ICP)现象是预富集低丰度生物样品最常用的方法之一。ICP会诱导出一个针对带电生物分子的非侵入性区域(即离子耗尽区),目标物可以在该区域边界处进行预富集。尽管ICP具有较高的预富集性能,但很难找到非传播性离子耗尽区的操作条件。为了克服这个狭窄的操作窗口,我们最近开发了一种用于时空固定预富集的新平台。与仅使用离子耗尽的先前方法不同,该平台还利用ICP的相反极性(即离子富集)来阻止离子耗尽区的传播。通过使富集区与耗尽区相对,这两个区域合并并停止。在本文中,我们描述了构建这个时空定义的ICP平台的详细实验方案,并通过与传统装置比较来表征新平台的预富集动力学。定性的离子浓度分布和电流-时间响应成功地捕捉到了合并后的ICP和单独的ICP之间的不同动力学。与传统方法只能在约5V时固定预富集位置不同,新平台可以在广泛的操作条件范围内(电压0.5 - 100V、离子强度1 - 100mM和pH 3.7 - 10.3)在特定位置产生目标浓缩栓。