数字微流控辅助将单个磁性粒子密封在飞升大小的反应孔中用于单分子检测。

Digital Microfluidics Assisted Sealing of Individual Magnetic Particles in Femtoliter-Sized Reaction Wells for Single-Molecule Detection.

作者信息

Decrop Deborah, Ruiz Elena Pérez, Kumar Phalguni Tewari, Tripodi Lisa, Kokalj Tadej, Lammertyn Jeroen

机构信息

MeBioS - Biosensors, Department of Biosystems, KU Leuven, Willem de Croylaan 42, 3000, Leuven, Belgium.

出版信息

Methods Mol Biol. 2017;1547:85-101. doi: 10.1007/978-1-4939-6734-6_7.

DOI:10.1007/978-1-4939-6734-6_7

PMID:28044289

Abstract

Digital microfluidics has emerged in the last years as a promising liquid handling technology for a variety of applications. Here, we describe in detail how to build up an electrowetting-on-dielectric-based digital microfluidic chip with unique advantages for performing single-molecule detection. We illustrate how superparamagnetic particles can be printed with very high loading efficiency (over 98 %) and single-particle resolution in the microwell array patterned in the Teflon-AF surface of the grounding plate of the chip. Finally, the potential of the device for its application to single-molecule detection is demonstrated by the ultrasensitive detection of the biotinylated enzyme β-Galactosidase captured on streptavidin-coated particles in the described platform.

摘要

近年来，数字微流控作为一种有前途的液体处理技术，已在各种应用中崭露头角。在此，我们详细描述了如何构建基于介电电泳的数字微流控芯片，该芯片在进行单分子检测方面具有独特优势。我们展示了如何在芯片接地极板的聚四氟乙烯 - AF表面图案化的微孔阵列中，以非常高的负载效率（超过98%）和单颗粒分辨率打印超顺磁性颗粒。最后，通过在所描述的平台中对捕获在链霉亲和素包被颗粒上的生物素化酶β - 半乳糖苷酶进行超灵敏检测，证明了该设备在单分子检测应用中的潜力。