- 學位論文
共平面電極之介電濕潤微流體系統在生醫檢測之應用與系統改良
Improvement of the Electrowetting-on-Dielectric Digital Microfluidic Systems and Its Biomedical Applications
摘要
近年來人類文明越趨先進與醫療科技之發達，生醫微機電(Bio- MEMS)正是微機電研究領域想要努力發展結合的重要目標。相較於傳統生醫檢測對於檢體體積需求量大、檢測耗時費工、容易汙染以及成本昂貴等等缺點，介電濕潤(Electrowetting on dielectric, EWOD)數位微流體晶片(Digital microfluidic chip)具有高效率、小體積、短時間、低成本、零污染之微生醫檢測優點。本研究中致力於以介電濕潤數位微流體系統進行各種生醫檢測，其中包括基因序列中單一核苷酸多型性(Single-nucleotide Polymorphism, SNP)之檢測、鼠精與卵細胞結合、以及鼠胚細胞之培養。此外我們致力於介電濕潤晶片之介電層品質改良，以克服介電濕潤晶片在操控生醫檢體時所面臨的最大困難 -- 介電崩潰。透過晶片製程改良，以低壓化學氣相沉積(LPCVD)之氮化矽(Si3N4)薄膜取代傳統電漿輔助化學氣相沈積(PECVD)氮化矽薄膜作為晶片介電層，能有效提升介電層崩潰電壓與操控生醫檢體時之穩定與持久性，使更多生醫檢測實驗能夠順利在晶片上完整呈現。 在生醫檢測之應用中，我們已成功利用共平面式電極之介電濕潤晶片進行分子生物學中去氧核醣核酸片段的接合反應(DNA ligation)。並且利用共平面式電極之介電濕潤晶片上蓋不需電極之特性，於其上加入白金微型加熱線圈裝置以整合溫度控制之功能。藉此提供檢測基因序列中單點核酸多形性(Single-nucleotide Polymorphism, SNP)反應中酵素作用適合的溫度環境，完成基因序列中單一核苷酸多型性之檢測。在生殖醫學(Reproductive medicine)方面，我們使用EWOD晶片作為體外授精、鼠胚發育的平台，研究介電濕潤檢體操控所施加的電場對於鼠胚、鼠卵的發育影響。本研究目前已夠成功在晶片平台上完成去氧核醣核酸片段接合、SNP檢測、鼠胚培養、鼠卵體外授精等等實驗。其結果顯示本研究中所使用之各種EWOD晶片具有良好的生物相容性以及優良的生醫檢體操控能力，對於未來EWOD系統在生醫檢測方面之應用有相當的貢獻。
並列摘要
Electrowetting-on-dielectric (EWOD) technique has been used in the digital microfluidic system (DMF). Droplets can be generated, transported, separated, and merged by EWOD electrodes actuation. The actuation of the droplets by EWOD electrodes have the features of precise droplet actuation, less contamination risk, reducing reagents volume, better reagents mixing efficiency, less reaction time, flexibility for integration with other elements and its portability, and therefore EWOD based DMF has been widely applied in biomedical or chemical fields as a powerful sample preparation platform. In this study, the coplanar electrodes EWOD system was used for different kinds of biomedical applications. We also focused on the quality improvement of dielectric layer of the EWOD chip for increasing the breakdown voltage during biomedical sample manipulation. We proposed a new EWOD device fabrication process using low pressure chemical vapor deposited (LPCVD) Si3N4 as dielectric layer. LPCVD Si3N4 exhibits not only high permittivity but also purity and uniformity, which increase the breakdown voltage of dielectric layer and make EWOD system more stable when applied in biomedical researches. DNA ligation and single-nucleotide polymorphism (SNP) detection in specific DNA sequence were performed by the EWOD platform. With the combination of a micro-heater on the top cap of the EWOD, biomedical reaction requires specific temperature can be done in the chip. To use the DMF for assisted reproductive technology (ART) researches, we also present an in vitro culture of mouse embryo and in vitro of mouse gametes by using the EWOD system. These biomedical applications were all successfully demonstrated by our EWOD digital microfluidics, the results suggest that the EWOD chip is biocompatible and has the potential for sample processing of molecular biology and assisted reproductive technology researches.
參考文獻
延伸閱讀
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