Laboratory of Nanostructures and Biosensing, Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
Lab Chip. 2012 Oct 21;12(20):3882-90. doi: 10.1039/c2lc40455a.
With recent advances in high-throughput proteomics and systems biology, there is a growing demand for new instruments that can precisely quantify a wide range of receptor-ligand binding kinetics in a high-throughput fashion. Here we demonstrate a surface plasmon resonance (SPR) imaging spectroscopy instrument capable of simultaneously extracting binding kinetics and affinities from 50 parallel microfluidic channels. The instrument utilizes large-area (~ cm(2)) metallic nanohole arrays as SPR sensing substrates and combines a broadband light source, a high-resolution imaging spectrometer and a low-noise CCD camera to extract spectral information from every channel in real time with a refractive index resolution of 7.7 × 10(-6) refractive index units. To demonstrate the utility of our instrument for quantifying a wide range of biomolecular interactions, each parallel microfluidic channel is coated with a biomimetic supported lipid membrane containing ganglioside (GM1) receptors. The binding kinetics of cholera toxin b (CTX-b) to GM1 are then measured in a single experiment from 50 channels. By combining the highly parallel microfluidic device with large-area periodic nanohole array chips, our SPR imaging spectrometer system enables high-throughput, label-free, real-time SPR biosensing, and its full-spectral imaging capability combined with nanohole arrays could enable integration of SPR imaging with concurrent surface-enhanced Raman spectroscopy.
随着高通量蛋白质组学和系统生物学的发展，人们对能够以高通量方式精确定量广泛的受体-配体结合动力学的新型仪器的需求不断增长。在这里，我们展示了一种能够同时从 50 个平行微流通道中提取结合动力学和亲和力的表面等离子体共振（SPR）成像光谱仪。该仪器利用大面积（约 cm(2))金属纳米孔阵列作为 SPR 传感基底，并结合宽带光源、高分辨率成像光谱仪和低噪声 CCD 相机，以 7.7×10(-6)折射率单位的折射率分辨率实时从每个通道提取光谱信息。为了证明我们的仪器在定量广泛的生物分子相互作用方面的实用性，每个平行微流通道都涂有包含神经节苷脂（GM1）受体的仿生支撑脂质膜。然后，在单个实验中从 50 个通道中测量霍乱毒素 b（CTX-b）与 GM1 的结合动力学。通过将高度并行的微流控装置与大面积周期性纳米孔阵列芯片相结合，我们的 SPR 成像光谱仪系统实现了高通量、无标记、实时 SPR 生物传感，其全光谱成像功能与纳米孔阵列相结合，可实现 SPR 成像与同时进行的表面增强拉曼光谱的集成。
