Doughan Samer, Noor M Omair, Han Yi, Krull Ulrich J
Chemical Sensors Group, Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, ON, L5L 1C6, Canada.
Methods Mol Biol. 2017;1571:301-326. doi: 10.1007/978-1-4939-6848-0_19.
Quantum dots (QDs) and upconverting nanoparticles (UCNPs) are luminescent nanoparticles (NPs) commonly used in bioassays and biosensors as resonance energy transfer (RET) donors. The narrow and tunable emissions of both QDs and UCNPs make them versatile RET donors that can be paired with a wide range of acceptors. Ratiometric signal processing that compares donor and acceptor emission in RET-based transduction offers improved precision, as it accounts for fluctuations in the absolute photoluminescence (PL) intensities of the donor and acceptor that can result from experimental and instrumental variations. Immobilizing NPs on a solid support avoids problems such as those that can arise with their aggregation in solution, and allows for facile layer-by-layer assembly of the interfacial chemistry. Paper is an attractive solid support for the development of point-of-care diagnostic assays given its ubiquity, low-cost, and intrinsic fluid transport by capillary action. Integration of nanomaterials with paper-based analytical devices (PADs) provides avenues to augment the analytical performance of PADs, given the unique optoelectronic properties of nanomaterials. Herein, we describe methodology for the development of PADs using QDs and UCNPs as RET donors for optical transduction of nucleic acid hybridization. Immobilization of green-emitting QDs (gQDs) on imidazole functionalized cellulose paper is described for use as RET donors with Cy3 molecular dye as acceptors for the detection of SMN1 gene fragment. We also describe the covalent immobilization of blue-emitting UCNPs on aldehyde modified cellulose paper for use as RET donors with orange-emitting QDs (oQDs) as acceptors for the detection of HPRT1 gene fragment. The data described herein is acquired using an epifluorescence microscope, and can also be collected using technology such as a typical electronic camera.
量子点（QDs）和上转换纳米粒子（UCNPs）是发光纳米粒子（NPs），在生物测定和生物传感器中常用作共振能量转移（RET）供体。量子点和上转换纳米粒子的窄发射光谱和可调节发射特性使它们成为多功能的RET供体，可与多种受体配对。基于RET转导的比率信号处理通过比较供体和受体发射来提高精度，因为它考虑了供体和受体绝对光致发光（PL）强度的波动，这些波动可能由实验和仪器变化引起。将纳米粒子固定在固体支持物上可避免诸如它们在溶液中聚集时可能出现的问题，并允许界面化学进行简便的逐层组装。鉴于纸的普遍性、低成本以及通过毛细管作用实现的固有流体传输，纸是用于即时诊断检测开发的有吸引力的固体支持物。鉴于纳米材料的独特光电特性，将纳米材料与纸基分析装置（PADs）集成提供了增强PADs分析性能的途径。在此，我们描述了使用量子点和上转换纳米粒子作为RET供体来进行核酸杂交光学转导的PADs开发方法。描述了将绿色发射量子点（gQDs）固定在咪唑功能化纤维素纸上，用作RET供体，以Cy3分子染料作为受体来检测SMN1基因片段。我们还描述了将蓝色发射上转换纳米粒子共价固定在醛修饰纤维素纸上，用作RET供体，以橙色发射量子点（oQDs）作为受体来检测HPRT1基因片段。本文所述数据是使用落射荧光显微镜获取的，也可以使用典型电子相机等技术收集。
