發展生物分子交互作用的奈米陣列光碟晶片

Development of the Nanoarray on a CD-based Chip for the Biomolecular Interaction

葉嘉仁(Chia-Jen Yeh)

指導教授 ： 林啟萬

國立臺灣大學/工學院/醫學工程學研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.03382

摘要

光碟式生物晶片發展行之有年，因具成本低廉、大量製造與提供離心力驅動微流體等優點；另外，只需要使用一般光碟機讀取信號的方式十分簡便與輕巧，使之在一般光學式生物晶片中極具競爭優勢。本論文首次提出使用沾筆式奈米微影技術(dip-pen nanolithography, DPN)在光碟片儲存資訊的軌道上大量且平行地製作奈米陣列，且利用其高精準空間定位優點使此奈米陣列的週期與軌道週期一致，使得奈米陣列檢測分子的容量趨近光碟記憶的容量，以期待未來能拓展檢測的分子種類。為了能夠達到低濃度的檢測，也利用光碟儲存資訊軌道的一維奈米結構進行鍍金膜並固定上奈米金粒子，探討此架構下區域性表面電漿共振(localized surface plasmon resonance, LSPR)的現象。
為了發展此新型奈米陣列光碟晶片的生物分子交互作用部分，首先在超平金上確認生物分子固定化流程。表面修飾streptavidin 的250 奈米螢光球已成功地固定在金膜上，並用原子力顯微鏡驗證。結果顯示此技術製程之晶片可用於抓取病毒顆粒的潛力。接著在不同容量的光碟片上鍍上50 奈米的金膜並固定上60 奈米的奈米金粒子，量取吸收光譜探討之間的LSPR效應。透過比較吸收光譜的特徵值，以藍光光碟片上的LSPR表現最顯著。最後，使用十二隻陣列式探針之DPN在光碟上平行地製程奈米陣列。奈米金粒子先與水凝膠混合，再用DPN平行地點在鍍金膜的光碟片上。透過擷取暗場顯微鏡影像與吸收光譜，水膠點陣列精準地排列在儲存軌道上，吸收光譜也顯示具LSPR的效應，可作為放大低濃度生物分子作用所用。
在本研究裡，首次使用DPN平行製程蛋白質奈米陣列並使用LSPR機制放大訊號的優點來發展生此新型物分子交互作用的奈米陣列光碟晶片，未來可結合微流道技術與改良光碟讀取數位訊息的方式朝實際應用邁進。

關鍵字

沾筆式奈米微影技術 ； 蛋白質陣列 ； 奈米金粒子 ； 光碟片 ； 區域性表面電漿共振

並列摘要

The CD-based biochip has been developed for several years, not only for its low cost, mass production and ability to provide the centrifugal force to drive micro-fluid; but also for its light weight and friendly usage which allows normal CD-ROM to read the signals. All of those advantages have made it more competitive among general bio-optical chips. In this research, we use dip pen nanolithography (DPN) to produce nanoarrays paralleled on the CD’s storage track, and take the advantages of its accuracy and precision to make the nanoarrays’ array consistent with the CD track period, so that to make the detection of molecules on nanoarray approaching the memory capacity of the disc, and look forward to expand the detection of molecular species. In order to reach a lower limit of detection, we also deposit gold thin film on one dimensional nanostructure of CD’s storage track and immobilize gold nanoparticles. We discuss the phenomenon of localized surface plasmon resonance (LSPR) under this framework.
In order to develop molecular interaction part of this novel of nanoarray CD-based biochip, we first recognized immobilization of biomolecules process on the ultra-flat gold. The 250 nm latex beads conjugated with streptavidin has successfully immobilized on the gold film, and verified by atomic force microscopy (AFM). The results show that this method has potential to be used in the process of capturing viral particles. Then, we also deposited 50 nm gold thin film on the disk’s storage track and immobilized 60 nm gold nanoparticles, and used absorption spectra to discuss the LSPR effect. We found the LSPR performance of Blu-ray discs is the most significant one by comparing the characteristics of absorption spectra. Gold nanoparticles mixed with hydrogel. Finally, we use twelve DPN probe arrays to make hydrogel nanoarray on the disc. We acquire image and absorption spectra of the hydrogel on the storage array by dark field microscope and spectroscope. The absorption spectra show the effect of LSPR provide the possibility to enhance the signal of detecting the low concentrations of biomolecules.
In this study, we used parallel DPN process in protein arrays and applied the advantage of LSPR to amplify the signal in this new type of CD-based chip for molecular interactions. The microfluidic technology and CD-ROM reading methods will be developed toward the practical application in the future.

並列關鍵字

dip-pen nanolithography (DPN) ； protein array ； gold nanoparticles ； compact disc ； localized surface plasmon resonance (LSPR)

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