Laboratory of Optical Imaging and Sensing, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China.
Biosens Bioelectron. 2011 Dec 15;30(1):180-7. doi: 10.1016/j.bios.2011.09.009. Epub 2011 Sep 18.
With the development of the microarray technology, demands for array detection techniques become higher and higher. For many microarrays, several biomolecular interactions occur simultaneously and the interplay of various factors that affect these interactions remains poorly understood. Detecting such interactions with a single technique can often be a difficult and complicated process. In this work we propose a combined technique which enables simultaneous angle-interrogation surface plasmon resonance (SPR) sensing and hyperspectral fluorescence imaging. This tandem technique offers two-dimensional imaging of the whole array plane. The refractive index information obtained from SPR sensing and the physicochemical properties obtained from fluorescence imaging provide a comprehensive analysis of biological events on the array-chip. In addition, SPR and fluorescence detection techniques confirm each other in experimental results to exclude false-positive or false-negative cases. In terms of SPR sensing performance, the refractive index resolution is 3.86×10(-6) refractive index units (RIU), and the detection limit is 10(4) cfu/ml of Escherichia coli bacteria. The resolving power and detection sensitivity of fluorescence imaging are approximately 20 μm and 0.61 fluors/μm(2), respectively. Finally, two model experiments, detecting the DNA hybridization and biotin-avidin interactions respectively, demonstrate the biomedical application of this system.
随着微阵列技术的发展，对阵列检测技术的需求越来越高。对于许多微阵列来说，几个生物分子相互作用同时发生，并且影响这些相互作用的各种因素的相互作用仍然知之甚少。使用单一技术检测这种相互作用通常是一个困难和复杂的过程。在这项工作中，我们提出了一种组合技术，该技术能够同时进行角度询问表面等离子体共振（SPR）传感和高光谱荧光成像。这种串联技术提供了整个阵列平面的二维成像。从 SPR 传感获得的折射率信息和从荧光成像获得的物理化学性质提供了对芯片上生物事件的综合分析。此外，SPR 和荧光检测技术在实验结果中相互证实，以排除假阳性或假阴性情况。在 SPR 传感性能方面，折射率分辨率为 3.86×10(-6)折射率单位（RIU），检测限为 10(4) cfu/ml 的大肠杆菌细菌。荧光成像的分辨率和检测灵敏度分别约为 20 μm 和 0.61 荧光/μm(2)。最后，两个模型实验，分别检测 DNA 杂交和生物素-亲和素相互作用，证明了该系统的生物医学应用。
