Hoettges Kai F, Hübner Yvonne, Broche Lionel M, Ogin Stephen L, Kass George E N, Hughes Michael P
School of Engineering, and School of Biomedical and Molecular Science, University of Surrey, Guildford, Surrey GU2 7XH, UK.
Anal Chem. 2008 Mar 15;80(6):2063-8. doi: 10.1021/ac702083g. Epub 2008 Feb 16.
Dielectrophoresis (DEP) offers many advantages over conventional cell assays such as flow cytometry and patch clamp techniques for assessing cell electrophysiology as a marker for cancer studies and drug interaction assessment. However, despite the advantages offered by DEP analysis, uptake has been low, remaining largely in the academic arena, due to the process of analysis being time-consuming, laborious, and ultimately allowing only serial analysis on small numbers of cells. In this paper we describe a new method of performing DEP analysis based on laminate manufacturing methods. These use a three-dimensional "well" structure, similar in size and pitch to conventional microtiter well plates, but offer electrodes along the inner surface to allow easy measurement of cell properties through the whole population. The result can then be determined rapidly using a conventional well-plate reader. The nature of the device means that many electrodes, each containing a separate sample, can be tested in parallel, while the mode of observation means that analysis can be combined with simultaneous measurement of conventional fluorimetric well-based assays. Here we benchmark the device against standard DEP assays, then show how such a device can be used to (a) rapidly determine the effects both of ion channel blockers on cancer cells and antibiotics on bacteria and (b) determine the properties of multiple subpopulations of cells within a well simultaneously.
与传统的细胞检测方法(如流式细胞术和膜片钳技术)相比,介电电泳(DEP)在评估细胞电生理方面具有诸多优势,可作为癌症研究和药物相互作用评估的标志物。然而,尽管DEP分析具有诸多优势,但其应用率却很低,主要仍局限于学术领域,这是因为分析过程耗时、费力,而且最终只能对少量细胞进行系列分析。在本文中,我们描述了一种基于层压制造方法进行DEP分析的新方法。这些方法使用三维“孔”结构,其尺寸和间距与传统的微孔板相似,但在内表面设有电极,以便能够轻松测量整群细胞的特性。然后,可以使用传统的微孔板读数器快速确定结果。该设备的特性意味着许多各自包含单独样品的电极可以并行测试,而观察模式意味着分析可以与基于荧光微孔板的传统检测的同步测量相结合。在这里,我们将该设备与标准DEP检测进行了对比,然后展示了这样的设备如何用于(a)快速确定离子通道阻滞剂对癌细胞和抗生素对细菌的影响,以及(b)同时确定孔内多个细胞亚群的特性。