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Real-time Full-spectral Imaging and Affinity Measurements from 50 Microfluidic Channels using Nanohole Surface Plasmon Resonance

机译：使用纳米孔表面等离子体共振从50个微流体通道的实时全光谱成像和亲和力测量

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摘要

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 extracting binding kinetics and affinities from 50 parallel microfluidic channels simultaneously. The instrument utilizes large-area (~cm2) 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. 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.
机译：随着高通量蛋白质组学和系统生物学的最新进展，对能够以高通量方式精确定量各种受体-配体结合动力学的新仪器的需求日益增长。在这里，我们展示了一种表面等离子体激元共振（SPR）成像光谱仪，能够同时从50个平行微流体通道中提取结合动力学和亲和力。该仪器利用大面积（〜cm 2 ）金属纳米孔阵列作为SPR传感基板，并结合了宽带光源，高分辨率成像光谱仪和低噪声CCD相机，以从中提取光谱信息。实时监控每个通道，折射率分辨率为7.7×10 -6 。为了证明我们的仪器可用于量化各种生物分子相互作用的实用性，每个平行的微流体通道都涂有含有神经节苷脂（GM1）受体的仿生支持脂质膜。然后在单个实验中从50个通道测量霍乱毒素b（CTX-b）与GM1的结合动力学。通过将高度平行的微流控设备与大面积周期性纳孔阵列芯片相结合，我们的SPR成像光谱仪系统可实现高通量，无标记的实时SPR生物传感，并且其全光谱成像功能与纳米孔阵列相结合可实现集成并发表面增强拉曼光谱的SPR成像技术

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