In-Situ Single-Molecule Fluorescence Imaging of Membrane Protein Interaction Using Nano-Structured Glass Plate

对于膜蛋白的原位单分子荧光成像，我们用计算光学模拟的方法研究了纳米光波导的结构，然后在玻璃衬底上制作了光波导。我们研究了与波导结构相关的光波导用于实时单分子成像的可行性。我们已经开发出制作蚀刻光波导和平面光波导的工艺。实验结果表明，刻蚀光波导能荧光检测蛋白质-蛋白质相互作用，并且光刻光波导信噪比是普通光波导信噪比的6倍。虽然我们还没有成功地利用Palner波导观察到人脑蛋白质的相互作用，但我们构建了观察的基本技术，并成功地利用平面波导探测了单分子荧光。

For in-situ single-molecule fluorescence imaging of membrane proteins, we investigated the structure of nano-waveguides using computational optics simulation, followed by fabrication of the waveguides on glass substrates. We studied the feasibility of the waveguides for real-time single-molecule imaging in relation to the waveguide structure. We have developped processes for the fabrication of both etched waveguides and planer waveguides. We showed that the protein-protein interaction can be fluorescently detected with the etched waveguide, and also that the signal-to-noise ratio of the etched waveguide is six times the conventional waveguide. Although we have not yet succeeded in observing menbrain protein interaction using the palner waveguide, we constructed the elemental techniques for the observation, and have succeeded in detecting the single-molecule fluorescence using the planer waveguide.