Department of Chemistry, University of Toronto, 80 St George St., Toronto, ON, Canada M5S 3H6; Donnelly Centre for Cellular and Biomolecular Research, 160 College St., Toronto, ON, Canada M5S 3E1.
Department of Chemistry, University of Toronto, 80 St George St., Toronto, ON, Canada M5S 3H6; Donnelly Centre for Cellular and Biomolecular Research, 160 College St., Toronto, ON, Canada M5S 3E1; Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College St., Toronto, ON, Canada M5S 3G9.
Biosens Bioelectron. 2016 Mar 15;77:845-52. doi: 10.1016/j.bios.2015.10.036. Epub 2015 Oct 22.
Electrochemiluminescence (ECL) is a sensitive analytical technique with great promise for biological applications, especially when combined with microfluidics. Here, we report the first integration of ECL with digital microfluidics (DMF). ECL detectors were fabricated into the ITO-coated top plates of DMF devices, allowing for the generation of light from electrically excited luminophores in sample droplets. The new system was characterized by making electrochemical and ECL measurements of soluble mixtures of tris(phenanthroline)ruthenium(II) and tripropylamine (TPA) solutions. The system was then validated by application to an oligonucleotide hybridization assay, using magnetic particles bearing 21-mer, deoxyribose analogues of the complement to microRNA-143 (miRNA-143). The system detects single nucleotide mismatches with high specificity, and has a limit of detection of 1.5 femtomoles. The system is capable of detecting miRNA-143 in cancer cell lysates, allowing for the discrimination between the MCF-7 (less aggressive) and MDA-MB-231 (more aggressive) cell lines. We propose that DMF-ECL represents a valuable new tool in the microfluidics toolbox for a wide variety of applications.
电化学发光（ECL）是一种灵敏的分析技术，在生物应用中具有很大的应用前景，特别是与微流控技术结合使用时。在这里，我们报告了电化学发光（ECL）与数字微流控（DMF）的首次集成。ECL 探测器被制作成 DMF 设备的 ITO 涂层顶板内，允许从电激发的发光体在样品液滴中产生光。通过对三（邻菲咯啉）钌（II）和三丙胺（TPA）溶液可溶性混合物进行电化学和 ECL 测量，对新系统进行了表征。然后，通过应用于带有 21 个碱基的脱氧核糖类似物的寡核苷酸杂交测定来验证该系统，该寡核苷酸是 microRNA-143（miRNA-143）的互补物。该系统具有高特异性检测单核苷酸错配的能力，检测限为 1.5 飞摩尔。该系统能够检测癌细胞裂解物中的 miRNA-143，能够区分 MCF-7（侵袭性较低）和 MDA-MB-231（侵袭性较高）细胞系。我们提出，DMF-ECL 代表了微流控工具箱中用于各种应用的有价值的新工具。
