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

为了实现膜蛋白的原位单分子荧光成像，我们利用计算光学模拟研究了纳米波导的结构，然后在玻璃衬底上制作了纳米波导。我们从波导结构的角度研究了该波导用于实时单分子成像的可行性。我们已经开发了蚀刻波导和平面波导的制造工艺。我们发现，用蚀刻波导可以荧光检测蛋白质-蛋白质相互作用，并且蚀刻波导的信噪比是传统波导的六倍。虽然我们还没有成功地使用帕尔纳波导观察到脑蛋白相互作用，但我们构建了基本的观察技术，并成功地使用平面波导检测到单分子荧光。

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.