被動式液滴定位免疫反應晶片之研究

本研究中以液滴為基礎的免疫檢測晶片開發於進行免疫檢測法的微陣列中，並且具有被動式液滴定位功能。使用微機電系統製造技術，微流道模仁結構被製造於印刷電路板(PCB)上，然後，倒入SYLGARD聚二甲基矽氧烷(PDMS)於模仁中，製造出微流道層，此外，利用電鍍將鎳鈷附著於PCB基板上，形成鎳鈷薄膜，目的是為了固定多種的組氨酸標籤蛋白，最後，藉由微流道層黏合於PCB基板上完成以液滴為基礎的免疫檢測晶片。為了測試這個提出基於液滴的免疫檢測晶片，使用腫瘤抑制蛋白p53和細胞外訊號有關的蛋白激酶（ERK1），並且矽油被選擇作為載流體。試劑在注入後還未形成液滴之前，先經過曲折的混合通道達到快速和徹底混合的效果，此後混合試劑抵達藉由懸掛的檔閥間隔的雙T字路口，並且受到載流體剪切，產生出兩種不同之液滴，這些液滴彼此不相互交融，因為矽油具有相對較高的粘度，單分散之液滴可以在一個較低的流速下產生。由於載流矽油流向排油流道，使每個液滴被輸送到檢測區孔洞完成定位，因此，組氨酸標籤蛋白被固定在鎳鈷層上於微陣列格式，以便後續進行免疫實驗和做螢光檢測。所檢測到的螢光強度也成濃度的比例封裝內容於在一個微小液滴之中，此提出的基於液滴的免疫檢測晶片允許操作順序的免疫檢測法通過利用液滴操縱自動進行。

關鍵字

液滴；定位

並列摘要

In this study, a droplet-based immunoassay chip having passive droplet positioning features was developed for performing immunoassays. Using MEMS fabrication techniques, the mold of the microfluidic channels was manufactured on the printed circuit board (PCB). Then, the microfluidic layer was formed by casting SYLGARD polydimethylsiloxane (PDMS) in the mold. In addition, a layer of Ni–Co film was coated on the PCB substrate by electrodeposition in order to immobilize the multiplex histidine-tagged proteins. Finally, the droplet-based immunoassay chip was produced by bonding the microchannel layer on the PCB substrate. To test this proposed droplet-based immunoassay chip, the tumor suppressor protein p53 and the extracellular-signal-related kinase 1 (ERK1) were used, and the silicon oil was chosen as the carrier fluid. When the reagents were injected, they passed through the zigzag mixing channel for rapid and thorough mixing. The mixture then arrived the double-T junction structured by a suspended diaphragm and sheared by the carrier flow, two different kinds of droplets were generated. These droplets were not fused with each other. The monodispersed droplets can be generated under a lower flow rate, since silicon oil has a higher viscosity. Each droplet was transported and positioned in a hole of detection area due to the flow of carrier oil flowing to the oil-drain channel. Thus, the histidine-tagged proteins were immobilized by the Ni–Co layer in microarray format for consequent immunoassay and fluorescence detection. The detected fluorescence intensity is proportioned to the concentration of the encapsulated content in a small droplet. This proposed droplet-based immunoassay chip allows the operation sequence of immunoassay being conducted automatically through the manipulation of droplets.

並列關鍵字

droplet ； positioning

參考文獻

[8] V. Srinivasan, V.K. Pamula, and R.B. Fair. “An integrated digital microfluidic lab-on-a-chip for clinical diagnostics on human physiological fluids,” Lab Chip, pp.310–315 , 2004 .

[1] A. Feuerborn, A. Prastowo, P.R. Cooka, and E. Walsh, “Merging drops in a Teflon tube, and transferring fluid between them, illustrated by protein crystallization and drug screening,” Lab Chip, pp.3766–3775, 2015.

[2] B. Gol, F. J. Tovar-Lopez, M. E. Kurdzinski, S.Y. Tang, P. Petersen, A. Mitchell, and K. Khoshmanesh,“Continuous transfer of liquid metal droplets across a fluid–fluid interface within an integrated microfluidic chip,” Lab Chip, pp.2476–2485, 2015.

[3] H. Wen, Y. Yu, G. Zhu, L. Jiang, and J. Qin, “A droplet microchip with substance exchange capability for the developmental study of C. elegans,” Lab Chip, pp.1905–1911, 2015.

[4] S. Ma, W.T.S. Huck, and S. Balabani, “Deformation of double emulsions under conditions of flow cytometry hydrodynamic focusing,” Lab Chip , 2015.

國際替代計量

被動式液滴定位免疫反應晶片之研究

全文下載

主題瀏覽