高速化多功光電生醫晶片儀之研發

Development of High Throughput Multi-functional Opto-electric Biochip System

莊叢優(Cong-You Jhuang)

指導教授 ： 李世光

共同指導教授 ： 吳文中

國立臺灣大學/工學院/工程科學及海洋工程學研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.01373

摘要

本研究成功的建構完成一套高速化多功光電生醫晶片檢測系統，並將此系統運用於薄膜厚度之量測與生物樣本之檢測上，此系統命名為Opto-BioMorph，簡稱OBMorph。本系統以橢偏儀檢測技術為主要開發之量測技術，可廣泛應用於高科技工業，如半導體製程及檢測設備、CD/DVD碟片，以及其他薄膜相關工程；在生物醫學的研究領域中，則可用來檢測病原或抗體、液固態介面的生物體動態結合分析、與表面濃度量測等。且本系統在量測生物醫學檢測速度上大幅提升，可做為生醫晶片與載具之研發及製造的重要工具。
本文中所建構之OBMorph檢測系統中，橢偏儀技術是採用相位調制架構設計，開發出利用拋物面反射鏡與球面反射鏡組合而成之控制入射角改變的光機，加上波板以完成相位調制動作；並以一參考光路做為校準光源之機制，以及設置流道系統提供生物檢測之用。控制角度變換機構採用控制精度0.4μm的伺服馬達來控制三角稜鏡的位置，配以拋物面鏡與球面鏡達到變換入射角度的功能，其入射角的精度可達±0.001°，入射範圍由20°至80°。另外，在決定系統角度時，以標準試片之布魯斯特角（Brewster angle）做為參考依據，提升角度之精確度。本架構中另一個特點是採用波板來進行調制相位動作，因為波板之相位延遲量較為穩定，較不受外界環境之影響，可有效效降低量測時間與提高系統精確度。而系統中所加入的參考光路，可根據此參考光路量測所得之光強進行校正並消除光源強度的不穩定性，亦可增加量測精度。由於以波板進行相位調制的功能，故可以達到高速化檢測之目的。
在進行薄膜厚度量測與生化反應量測時，必須先利用系統之轉換函數來校正與分析橢偏參數，提升其整體的精確度。嘗試以平面反射鏡、鍍厚膜鉻之矽晶圓、以及純矽晶圓作為系統較準的標準試片，實驗所得之結果發現以矽晶圓所得之系統參數為基準，量測並計算所得之膜厚與商用量測儀器之結果最相近，誤差僅為3%。在生化反應量測中，以酵素連結免疫分析反應(enzyme-link immunosorbent assay, ELISA)作為量測之基本生化反應流程，從量測結果之數據圖可知，在生物反應薄膜檢測應用上，OBMorph顯現出極高之分辨力。本論文研究的實驗結果與理論值之相契合度，成功證實了OBMorph在橢偏儀功能上擁有良好的解析度，且亦達到高速化之功能。在生物檢測方面，OBMorph的流道子系統之設置，可使系統擁有量測生物動態鍵結反應的能力，證實OBMorph在生醫檢測上之潛力。

關鍵字

橢偏術 ； 表面電漿共振

並列摘要

In this dissertation, a high throughput multi-functional opto-electric biochip system was built. The system named OBMorph, which is an acronym of Opto-bioMorphin, was designed to analyze the biomolecular interaction on a protein biochip. This instrument based on ellipsometry theory can be utilized in high-tech industries, which may include semiconductor manufacturing, inspec-tion equipment, CD/DVD disks, and other thin-film related operations. In addi-tion, application to biomedical area can include film thickness, physical parame-ters, concentration, mass, density, kinetic constants, binding specificity, and so on. By using the wave-plate to modulate phase retardane, the bio-tech detection ac-curacy can be significantly improved. The testing data obtained clearly indicate that OBMorph system is able to undertake accurate measurement for biomedical applications.
OBMorph adopted a paraboloidal mirror to vary the incident angle, the wave-plates to modulate phase retardation, a reference optical path to stabilize the optical source, and a flow-injection system to drive bio-sample into the bio-chip. In order to having high precision in the ellipsometry subsystem of OB-Morph, it adopt a 0.4μm accuracy servomotor to control the stage that carried the right angle mirror and to control the variation of the incident angle. By using this method, a ±0.001°angular control accuracy between 20°to 80°can be achieved. The interrogation angle could be retrieved accurately which leads to much higher sensitivity and stability by identifying Brewster’s angle of the stan-dard tester. This biosensor developed in this thesis is phase modulated ellipsome-try, which utilized a wave-plate to modulate phase retardance. By using this wave plate modulation method, the phase retardance dectected becomes more stable when compared to that of the piezoelectric actuator driven method. In our system, a reference optical path would be carried out, and the light souce in-stability would be reduced with the feedback of the reference beam. With above mentioned arrangements, OBMorph could serve a research and a high throughput measurement tool in biomedical measurement.
In the procedures of detecting the bio reactions, the transfer function of sys-tem must be applied to calibrate and analysis the ellipsometry parameters of the samples and this will improve the accuracy. The commercial reflected mirror, the bare silicon wafer, and the wafer coated with 300 nm chromium film were taken as the standard testers. As the experimental results, the error is about 3% of the measured gold thickness which is calculated by the system function ac-cording the bare silicon wafer. After that, the biomolecular interactions called enzyme-link immunosorbent assay, ELISA have been measured. OBMorph shows good ability to distinguish every step of the reactions. Since the experi-mental results and the simulation data agree well with each other, it is proven that ellipsometer possesses both great resolution and high throughput within OB-Morph. Furthermore, this system is equipped with high-efficiency flow injec-tion system that can measure not only the immobilized monolayer of IgG and bio-linker but also the thickness of bio layers. In summary, both the simulation and the experimental results obtained proved that OBMorph has many advan-tages in the bio-molecular measurement arena.

並列關鍵字

ellipsometry ； surface plasmon resonance

參考文獻

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