本論文主要研究表面電漿子與粒子電漿子之局域電磁場強化現象於表面強化拉曼散射(surface-enhanced Raman scattering，SERS)與螢光影像技術的影響。由於生物分子之拉曼訊號相當微弱，必須藉由粒子電漿子的操控使訊號放大。採用兩種SERS基材來觀察生物分子之拉曼訊號，一種為膠體奈米銀粒子，另一種則使用飛秒雷射加工製造。第一種方式是利用化學合成方式來製造奈米銀粒子，用來排列於奈米膜層上之金屬粒子大小與其分佈情形，並建立聚焦式與衰減全反射方式微拉曼光譜儀來加以量測分析；第二種則為使用飛秒雷射加工矽晶圓製造SERS的基材，基材能同時產生類似光柵的奈米結構及藉由雷射誘發光還原效應形成奈米銀粒子，可達單一步驟且大面積的製作，其訊號增強因子可達106倍。SERS基材將可研究固定於表面之生物分子結構資訊，得到其SERS光譜訊號並加以分析討論，我們將可建構一藉由電漿子來強化量測訊號之SERS的生物分子辨識平台，提供生物分子的結構改變之資訊，以期在生物分子結構的研究上有所幫助。
另一方面則利用表面電漿子來增強雙光子激發螢光之影像，藉由Fresnel方程式和古典偶極輻射模型理論研究局域電場增強、量子效率及螢光發射耦合效率與螢光分子與金屬表面之間之表面電漿子效應。經由局域電場增強與螢光發射耦合效率理論分析得到，銀膜的厚度在40 nm時，有最佳的雙光子激發螢光收光效率。而且濺鍍銀膜前先鍍上一層鍺層，將可改善銀膜的表面粗糙度低於1 nm，這將可減少局域熱點的產生，其中局域熱點會破壞影像均勻度。而在高能雷射的照射下，發現將會破壞原有設計的膜層結構，藉由鍍上二氧化矽的保護層來避免銀膜被破壞的情形，還可降低銀膜對螢光的淬滅效應。在螢光影像上，已取得螢光球在水中的雙光子激發之螢光影像及其後焦平面的表面電漿耦合放光環之影像，這樣的膜層設計將可有效率的用來觀測水中的螢光影像，進而可應用在生物分子螢光影像觀測的研究方面。

In this thesis, local electric field enhancement via surface plasmons (SPs) and particle plasmons (PPs) has been investigated in the influences of surface-enhanced Raman scattering (SERS) and fluorescence imaging. The Raman signal of biomolecules is tiny, and hence it is needed to be magnified by other approaches such as manipulating SPs and PPs. Two kinds of SERS substrates were used for biomolecule observation. One is to control colloid nanoparticles and the other is fabricated by femtosecond laser. Using a chemical synthesis approach to fabricate silver (Ag) nanoparticles, the size and distribution of embedded metal nanoparticles can be controlled on the sensor surface to enhance the local electric field. On the other hand, the silicon substrate in the silver nitrate solutions was machined by the femtosecond laser fabrication. The substrate can achieve simultaneously the generation of grating-like nanostructures and the formation of Ag nanoparticles on the surface via the laser-induced photoreduction effect. By femtosecond laser fabricated substrate, the enhancement factor can reach to 106. The biomolecular recognition platform utilizing the plasmon-enhanced signal of SERS offers the structural information of biomolecules.
By utilizing the Fresnel equation and classical dipole radiation modeling, local electric field enhancement, fluorescence quantum yield, and fluorescence emission coupling yield via SPs were theoretically analyzed between the fluorescence dye and the metal film. The optimal condition of an Ag film deposited on a cover slip for surface plasmon-coupled emission (SPCE) induced two-photon excited fluorescence (TPEF) based on an objective-based, total internal reflection (TIR) microscope was investigated. According to the theoretical simulations of local electric field enhancement and fluorescence coupled emission efficiency, the thickness of the Ag film should be about 40 nm in order to maximize the TPEF collection efficiency by the objective. The deposited Ag film with a germanium seed layer on a cover slip exhibits additional improvement in surface smoothness by reducing variations in surface roughness to below 1.0 nm, thereby reduces local hot spots which degrade the image uniformity. Moreover, an Ag film with a 20 nm-thick SiO2 spacer not only prevents damage caused through interaction with the aqueous solution under high laser power irradiance, but also reduces the fluorescence quenching effect by the Ag film. By optimizing the Ag film thickness, surface smoothness, and a protective dielectric spacer, efficient TIR TPEF imaging can be achieved through SPCE.

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