薄層聚合酵素連鎖反應系統與螢光檢測系統之開發

Abstract

本論文研究薄層聚合酵素連鎖反應(PCR)系統及螢光檢測系統，薄層聚合酵素連鎖反應系統之反應槽以PDMS做為材質，並且以單一鑄模方式製作，製程簡單且具有高製造良率。昇降溫部份以熱電致冷晶片做為加熱元件，搭配離心風扇做強制對流泠却。溫控部份以LabVIEW軟體擷取Thermocouple量測的溫度資料，再進行PID控制以改變TEC的加熱功率輸出。經實驗證明此薄層聚合酵素連鎖系統可成功的複製DNA序列。螢光檢測系統分為二部份裝置，雷射誘導螢光檢測部份以532nm 波長的綠光半導體雷射做為激發光源，搭配適當光學元件，最後以光電倍增管偵測樣本螢光訊號，並以 LabVIEW 軟體程式做為資料擷取系統。螢光影像系統則以UV光做為DNA染料激發光源，再以CCD擷取螢光影像。本螢光檢測系統利用樣本步進平台可作定點或掃描之螢光偵測，達成多用型螢光檢測系統之目的，實驗證實可成功的偵測DNA螢光訊號。

A thin-film Polymerase Chain Reaction (TF-PCR) system and a fluorescence detection system have been developed. Made of PDMS, the TF-PCR is fabricated by casting with easy of manufacturing and very high quality. The TF-PCR contains thermoelectric cooling components (TEC) as heaters, and uses a centrifuge system fan for forced convention cooling. The temperature controlling system is composed of a thermocouple as a temperature sensor, a DAQ card gather to voltage signals and a PC with the LabVIEW program. The monitoring and controlling of the TEC heater is through a PID program. Successful amplification of DNA strains of a shrimp virus with 120 and 226 base pair has been demonstrated by this TF-PCR. The fluorescence detection system can be divided into two subsystems. The LASER-induced fluorescence detecting subsystem includes a Green color semiconductor LASER (532nm in wavelength) as an exciting light source, followed by proper optical components and a photo multiplying tube (PMT), then fluorescence signals from DNA samples can be caught. The fluorescence image subsystem has a UV light source and can extract images from a CCD camera. This fluorescence detection system also uses step motors to control the sample stand, so that fixed scanning or moved scanning is possible. This fluorescence detection system is a multiple functional detecting system.