整合微流體系統與表面電漿光柵晶片於醣化血紅素量測

On-Chip HbA1c Detection Utilizing a Compact Surface Plasmon Grating Sensor Integrated on a Microfluidic System

周煥庭(Huan-Ting Chou)

指導教授 ： 李明昌

國立清華大學/電機資訊學院/光電工程研究所/碩士(2014年)

摘要

表面電漿生物感測技術擁有比其他生物感測方法有更高的靈敏度，如今已經發展出很多不同的表面電漿生物感測方法。本論文主要是利用光柵式表面電漿感測方法偵測生物檢體。此方法最主要是藉由觀察反射頻譜的變化量，而去感測光柵表面物質的變異。我們利用有限時域分析法 (Finite Difference Time Domain)這個方法去模擬後發現改變光柵表面形貌會影響反射頻譜的特性。接著我們將表面電漿晶片整合在微流道上利用微流道能夠大幅減少檢體用量和使液體傳輸自動化的特性使整個偵測流程更加快速方便。再來我們利用整合微流道的表面電漿晶片偵測糖尿病的指標HbA1c 生物檢體。一開始我們先利用螢光標定的方式確認第一層的適體(aptamer)有最好的自組裝，確認好最佳濃度和時間後，我們分別通入0~3 μg/μl 的HbA1c 接著在通入0.025 μg/μl 糖化血紅素抗體(HbA1c antibody)，而我們可以看到HbA1c的訊號偏移量會隨著濃度改變而有不同、而HbA1c antibody 雖然是注入的相同濃度，但會因為HbA1c 的鍵結量不同而有線性變化。

關鍵字

表面電漿 ； Aptamer

並列摘要

Surface plasmon utilized for biosensing applications has been demonstrated with high sensitivity. There are many types of surface plasmon biosensors today. In this research, we focus on a grating-based surface plasmon biosensor. The grating-based SP biosensor detects the biomolecule through interrogating the shift of the reflection spectrum. According to the simulation result by Finite Difference Time Domain (FTDT) method, we found that the bandwidth is generally related to the profile of the grating. In addition, we integrate microfluidic systems into the SPR chip. Microfluidic systems have the advantage of reducing the usage of specimen and facilitating the delivery of liquid through the entire detection process quickly and easily. After optimizing the concentration and process time of applying HbA1c-specific aptamers, we passed through 0~3 μg/μl of HbA1c, followed by injecting 0.025 μg/μl of HbA1c antibody. We observed the SPR wavelength shifts along with the concentrations of HbA1c, showing a good linear correlation. We also demonstrated that the detection system can be miniaturized for a portable apparatus.

並列關鍵字

表面電漿 ； 適體

參考文獻

[19] 吳民耀 and 劉威志, "表面電漿子理論與模擬," 物理, vol. 28卷, pp. 頁486-496, 200604 2006.

[15] 楊自森, 吳見明, 蔡旻龍, 莊淳宇, 崔豫笳, and 許志楧, "分子生醫光電科學與技術," 物理, vol. 27卷, pp. 頁670-686, 200510 2005.

[1] C. J. Huang, H. I. Lin, S. C. Shiesh, and G. B. Lee, "An integrated microfluidic system for rapid screening of alpha-fetoprotein-specific aptamers," Biosens Bioelectron, vol. 35, pp. 50-5, May 15 2012.

[2] Y. Yang, D. Yang, H. J. Schluesener, and Z. Zhang, "Advances in SELEX and application of aptamers in the central nervous system," Biomol Eng, vol. 24, pp. 583-92, Dec 2007.

[3] D. H. Bunka and P. G. Stockley, "Aptamers come of age - at last," Nat Rev Microbiol, vol. 4, pp. 588-96, Aug 2006.