臺灣博碩士論文加值系統

以離子風誘導之微型漩渦將液體中的懸浮微粒聚集以達濃縮效果，此技術為利用針尖狀電極架設在離子風晶片上方並施加高頻高壓電場(頻率60kHz、600-700VRMS)，此時電極會將其周圍的空氣粒子極化並使之帶有電性，當帶電粒子累積於電極尖端達飽和時，會因同性電相斥使帶電粒子排出形成離子風。本研究以離子風誘導之微型吹乾濃縮系統並結合微型電熱裝置。離子風裝置會吹拂晶片上的液滴產生一道剪切作用力，此作用力推動液體表面轉動，使整個液滴旋轉產生漩渦，且同時吹乾液體，此裝置搭配加熱系統可以加速液滴的蒸發，使得較大體積液滴中的懸浮微粒將會快速聚集並吹乾後濃縮。本實驗利用具有表面增強拉曼散射標籤(Nanoaggregate Embedded Beads , NAEBs)的沙門氏菌樣品液滴(NAEB@ Salmonella)，經由離子風系統聚集並結合電熱裝置使之加速液滴內部的熱對流以增加揮發效率。為了達到優化條件，先使用5µm的聚苯乙烯微粒進行模擬，探討加熱所需的條件，接著濃縮較大樣品體積如100µL的樣品觀察其結果。當單純使用先前的離子風裝置時，吹乾濃縮時間大約為一個多小時，若搭配加熱系統，可使液滴加速吹乾並增加濃縮效率，縮短吹乾濃縮時間至約30分鐘，再利用拉曼分析儀針對濃縮點進行檢測並驗證沙門氏菌的存在，實驗結果顯示偵測極限達到10 CFU/mL，具備檢測單一菌體的能力。最後，我們將此系統裝置應用在模擬真實樣品(美生菜、冰塊)。此裝置系統具有操作容易、樣品快速濃縮，搭配表面增強拉曼標籤提高偵測靈敏度，可讓此系統成為食品病原菌的快速檢測平台。

This study is aimed to integrate thermoelectric heating unit with ionic wind-based miniaturized evaporation device to quickly concentrate food pathogen sample containing Salmonella, bound with Raman tags to determine.
Surrounding gas molecules near one metallic needle applied with high voltage can be ionized to accumulate. These charge species finally repel each other to eject one gas stream i.e. ionic wind from the needle tip. Previously we use this ionic wind swiping across the top of one droplet to create a centrifugal vortice to trap Raman-tag bound bacteria samples. At the same time, ionic winds accelerate droplet evaporation to achieve 1000-fold concentration effect. Single copy determination of Salmonella in 10 micro-L sample has been reported by our group using this device. (The voltage 600 VRMS of high frequency 60 kHz is used for safety concern.) In this study, thermoelectric heating unit is imbed in the aforementioned ionic wind concentration device to further improve drying efficiency to handle real samples of larger volumes.
Using the ionic wind-based concentration chip containing heating unit, the thermal convection in the sample droplet is accelerated to efficiently increase liquid evaporation rate. We demonstrate the drying and concentration time of 100 micro-L sample of Salmonella are reduced from close to two hours without using heating device to approximately 30 minutes. Raman tag-bound Salmonella are successfully determined at the level of 10 CFU/mL, close to the requirement to detect ready-to-eat food samples, of which the safety standard is zero tolerance. In the future, we will evaluate the feasibility of this device to detect Salmonella in spiked real samples including ice and vegetable.

摘要 I
Abstract III
目錄 V
圖目錄 VIII
表目錄 XIV
第1章 緒論 1
1.1. 前言 1
1.2. 研究目的 2
1.3. 實驗原理 3
1.3.1. 離子風文獻回顧 3
1.3.2. 離子風原理 5
1.3.3. 沙門氏菌(Salmonella) 9
1.3.4. 拉曼光譜與拉曼散射[7] 13
1.3.5. 表面增強拉曼散射(Surface-enhanced Raman scattering, SERS) 16
1.3.6. 奈米聚集團粒子拉曼標籤(Nanoaggregate-embedded beads, NAEB) [13] 20
1.3.7. 熱電致冷晶片(Thermoelectric Cooling Module) 22
1.3.8. 革蘭氏染色法 (Gram Staining) [15] 24
第2章 材料與方法 27
2.1. 實驗材料與樣品 27
2.1.1. 實驗儀器與材料 27
2.1.2. 實驗樣品與藥品 29
2.2. 樣品前處理及配製 30
2.3. 離子風晶片 36
2.4. 實驗系統裝置與條件設定 37
2.5. 實驗流程 41
2.6. 革蘭氏染色實驗步驟 42
第3章 結果與討論 43
3.1. 微型電熱裝置的最佳條件 43
3.1.1. 探討加熱方式搭配離子風系統 43
3.1.2. 使用電熱元件加熱的最佳條件 47
3.2. 利用離子風暨微型電熱裝置微濃縮模擬NAEBs與沙門氏菌反應樣品與方法確認 52
3.2.1. 離子風暨微型電熱裝置微濃縮吹乾現象 52
3.2.2. 微濃縮與吹乾連續稀釋的NAEB@Salmonella樣品 53
3.2.3. 連續稀釋的NAEB@Salmonella樣品拉曼光譜 54
3.2.4. 微濃縮聚集與偵測結果比較 58
3.2.5. 革蘭氏染色法應用於沙門氏菌之實驗結果 59
3.3. 利用離子風暨微型電熱裝置模擬大體積樣品的最佳實驗條件 60
3.3.1. 離子風系統對粒子大小選擇性 60
3.3.2. 模擬的高濃度樣品最佳實驗條件 61
3.3.3. 模擬大體積樣品中低濃度沙門氏菌最佳實驗條件 68
3.4. 模擬真實樣品市售冰塊的檢測 73
第4章 結論與未來展望 79
參考文獻 80
附錄 模擬真實樣品市售美生菜的檢測 83

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