分歧微流道與微透鏡之設計及其在光碟酵素連結免疫分析之應用

在臨床醫學上有許多免疫診斷的方法，當中一種常用的檢測技術為酵素免疫分析法(Enzyme-Linked Immunosorbent Assay, ELISA)，現今利用微機電製程技術已進階發展成光碟酵素免疫分析法(Compact Disk Enzyme-Linked Immunosorbent Assay, CD-ELISA)，此種檢測技術有試劑量少、實驗時間短等優點，而跟傳統的ELISA設備相比較，CD-ELISA的設備為桌上型較為輕便，且配合光學模組可使檢測精度提高，因此具有廣泛應用在免疫診斷的潛力。本研究利用微機電製程技術製作分歧微流道Y型與U型以及利用PDMS製作微透鏡，且能應用在光碟酵素免疫分析法上。在分歧微流道部份，先以有限元素分析軟體COMSOL模擬能使微流道流量平均分流的設計，再藉由黃光微影製程製作出SU-8材質的母膜，並以PDMS翻模使母膜能重複使用，接著以氧電漿接合技術將PDMS製成的微流道與光碟片做接合。而在微透鏡方面是將聚焦長度做系統性的分析並使用光學軟體ZEMAX模擬，再透過實驗加以驗證。最後在免疫分析檢測時，整合微透鏡的光碟微流體晶片可用來提升其生物檢測時反應訊號。實驗結果顯示整合微透鏡在光碟微流體晶片上後的檢測效果能大幅的增強原本的檢測訊號提升其靈敏度。

關鍵字

光碟酵素免疫分析法；微流道；微透鏡；生物檢測

並列摘要

Enzyme-linked immunosorbent assay (ELISA) microchips have numerous advantages over existing technologies. However, the low reagent volume in such chips results in a limited dynamic linear range, a low signal-to-noise ratio, and a poor detection sensitivity. Therefore, there is an urgent need for simple and efficient methods to enhance the sensitivity of ELISA assays performed using polymeric microfluidic devices. In this study, a CD-chip with splitter microfluidic channel has fabricated utilizing MEMS technology. A finite element analysis tool (COMSOL) has been established to analyze the flow of the splitter microfluidic channel. A novel device comprising polydimethyl-siloxane (PDMS) microlenses bonded to a microfluidic compact disk (CD) is proposed for ELISA applications. The PDMS microlenses are fabricated using a simple soft replica molding method and are bonded to the microfluidic CD using oxygen plasma treatment technique. A commercial software tool (ZEMAX) has been established to analyze the focal length of the microlens. A laser-induced fluorescence (LIF) bio-detection system consisting of the integrated microfluidic CD/PDMS microlenses and an optical detection module is constructed and is used to examine the enzymatic reaction of HPPA. The experimental results show that the PDMS microlens focusing effect yields a significant improvement in the intensity of the detected fluorescence signals. As a result, the proposed device represents an ideal solution for ELISA immunoassays and other high-sensitivity bio-detection applications.

並列關鍵字

Bio-detection ； ELISA ； Microchannel ； Microlens ； Splitter

參考文獻

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分歧微流道與微透鏡之設計及其在光碟酵素連結免疫分析之應用

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