近年來人們在工作、日常生活、休閒娛樂上都大量使用3C電子產品，眼睛長期疲勞致使眼角膜的健康以及能抑制細菌滋長的淚液分泌狀況不佳。許多人又因為美觀與便利性而習慣地佩戴隱形眼鏡，反覆穿戴隱形眼鏡會使眼角膜磨損，如再加上隱形眼鏡的清潔方式不當，角膜便容易受到細菌的感染，延誤治療將導致急性角膜炎、角膜潰瘍、角膜穿孔，輕則視力下降，嚴重則導致失明。傳統檢測方式主要為透過臨床問診推斷及病灶採樣的培養與分析，但其過程非常耗時。故此研究欲提供一種新的檢測方式，在臨床上能更快、更準確的分析感染的微生物種類，使醫生能提供病患更即時的治療。此篇欲利用拉曼光譜儀的快速檢測以及靈敏度高的特性，作為一種快速檢測臨床微生物指紋的工具。本研究主要分為四個部分：(1) 利用銀鏡反應在濾材上附著銀奈米粒子，使之成為具有拉曼增顯以及過濾濃縮效果的表面增顯拉曼散射過濾薄膜(SERS Filter)，並探討其參數最佳化及過濾濃縮的效果。(2) 利用SERS Filter來對多種標準菌種、臨床菌種以及不同編號的純菌進行拉曼檢測，以獲得多種菌種的拉曼指紋圖譜，並進一步利用主成份分析法(PCA)將獲得的指紋圖譜轉換成PCA分群圖，形成一種可以用來快速判斷菌種的分群資料庫。(3) 針對低濃度樣本的檢測時，加入AuNPs可與待測物在三維空間有更多的接觸，給予更強的增顯訊號，於此部分首先探討AuNPs的型態變化對化學分子檢測的參數最佳化。(4) 將AuNPs對細菌的檢測參數進行優化，並將兩種拉曼增顯基材結合，成為雙重增顯基材(AuNPs/ SERS Filter)，透過對低濃度細菌樣本的過濾濃縮，使細菌與AuNPs大量累積於SERS Filter表面，使得低濃度樣本也能獲得清楚的指紋訊號。此篇研究的雙重增顯基材其成本低且能快速製備，透過微生物指紋圖譜與PCA分析技術結合，可快速進行微生物菌種的判斷。在未來發展上，大量建立微生物指紋圖譜資料庫，便可利用於臨床醫學、環境汙染以及食安問題的第一線快速檢測。 In the recent years, number of ophthamology disorders and diseases are increased clinically which strong correlation to the usage of electronic 3C devices. Contact lenses, another important risk, provides not only eyepower correction ability but also acts as cosmetic products. However, contact lens wearing can cause a variety of problems ranging from mild to severe such as dry eyes, allergic eye disease, distortion of the cornea, blood vessels growing in the cornea due to a lack of oxygen, scratches or abrasions on the cornea, and cornea infection which can lead to blindness if the infection is severe.The goal for this thesis was to prepare nano-scale metallic materias for rapid and high sensitivitybacteris Raman and surface enhanced Raman spectroscopy (SERS) ananlysis. The specific bacteria Raman and SERS fingerprints were further classified under principle components analysis (PCA) which performed bacteria groupping ability and can be applied for further clinical applications. This study included fourparts:1. Preperation of silver nanoparticles decorated SERS Filter membranes were used for bacteria SERS fingerprint measurement and the filtration and concentration functions were also studied.2. SERS measurement and analysis of clinical isolated bacteria were performed and compared with standard strans. PCA was performed for rapid classification and characterization of bacteria between strains and specises.3. Gold nanoparticles (AuNPs) was synthsized for direct contect with target bacteria in three dimention. Chemical analyte was performed to stufy signal enhancement ability of nanoparticles prior to bacteria measurement. Size and morphology of particles were discussed. 4. Double enhancement were performed by intergration of metallic nanoparticles (AuNPs) and SERS substrate. Low concentration samples were be tested for understand the concentration ability of prepared SERS Filter. This thesis presented a rapid, high sensitive and specificity tool based on double enhancement SERS technique for low concentration bacteria SERS measurement by combination of AuNPs and SERS Filter. PCA analysis results showed clinical isolated bacteria from keratitis specements can be classified succefully under strains level. The method reported here not only for clinical microorganism screening, but also can be applied for environments and food safety monitoing.
