奈米電漿子整合自動化微流道控制系統進行免標定即時多重多工細胞激素檢測

A Nanoplasmonic-integrated Automatic Microfluidics Control System for Label-free, Real-time, Multi-parallel and Multiplex Cytokine Detection

陳志翔(Jhih-Siang Chen)

指導教授 ： 黃念祖

國立臺灣大學/電機資訊學院/生醫電子與資訊學研究所/碩士(2020年)

https://doi.org/10.6342/NTU202000515

摘要

細胞激素作為免疫系統中重要的溝通橋樑，能調節免疫反應，且其與發炎與感染反應有高度相關，故細胞激素的種類及濃度變化可用於觀察人體即時的免疫系統狀態。然而，現今標準的細胞激素檢測技術因其需要使用螢光標定檢測，往往需要大量的樣本體積量、繁複的的標定程序及冗長的反應時間，使其只能量測最後反應的濃度值，無法即時觀察隨著療程而變化的狀態。為了突破上述限制，免標定檢測技術在近幾十年來時常被使用作為細胞激素檢測的平台，而相較於其他類型的免標定檢測技術，光學式的感測器，例如奈米電漿子感測器，則因為其無須複雜系統架設及高靈敏度特性受到相當高的關注，此外將奈米電漿子整合在微流道之裝置還有微量化樣品量、簡化繁複的樣品準備過程、避免樣本損耗甚至能縮短檢測時間的優勢。
在本篇論文中，我們提出兩種製程方式包含快速熱退火及雷射雙光干涉微影結合奈米壓印技術以製作大面積奈米電漿子生物感測器(約平方公分)，另外，我們也將自動化微流道裝置、客製化光學平台與奈米電漿子生物感測器整合，用以進行免標定、多重與多工細胞激素檢測同時避免潛在操作錯誤及勞力密集之標定步驟。我們相信此系統能應用於多種生物分子檢測並且充分顯示具有研究細胞表型及疾病診斷之潛力。

關鍵字

微流道 ； 奈米電漿子 ； 細胞激素 ； 多重檢測 ； 多工檢測

並列摘要

Cytokines are important bridges in the immune system and regulate the immune response, which are highly related to inflammation and infection. Hence, cytokine detection is a perfect index for observing the immune system. However, the current golden standard techniques for cytokine detection not only require large sample volume and labor-intensive labeling processes but also time-consuming and even readout end-point results. To eliminate these limitations, label-free sensors have been applied for cytokine detection over the decades. Compared to other label-free biosensors, optical sensors such as nanoplasmonic sensors receive lots of attention due to its simple system setup and high sensitivity nature. Moreover, nanoplasmonic-integrated microfluidics takes advantage of combining many features in one chip for minimizing the sample volume, simplifying the complicated sample preparation, preventing the sample loss and even shortening the detection time.
In this thesis, we propose two methods to fabricate large-scale nanoplasmonic biosensor (about cm2), rapid thermal annealing (RTA) and laser interference lithography (LIL) combing nanoimprint lithography (NIL). Furthermore, an automated microfluidics control system and a customized optical platform are integrated with the nanoplasmonic biosensor for label-free, multi-parallel and multiplex cytokine detection to avoid any potential operational error and labor-intensive labeling process. We believe this system can be applied for multiple biomarkers detection, which shows the great potential in studying cellular phenotyping and disease diagnosis.

並列關鍵字

Microfluidics ； Nanoplasmonic ； Cytokines ； Multi-parallel detection ； Multiplex detection

參考文獻

1. Bakri Hassan, A., et al., Increased Serum Levels of Immunoglobulins, C-reactive Protein, Type 1 and Type 2 Cytokines in Patients with Mixed Connective Tissue Disease. Journal of Autoimmunity, 1998. 11(5): p. 503-508.

2. Ng, A., et al., IL-1β, IL-6, TNF- α and CRP in Elderly Patients with Depression or Alzheimer’s disease: Systematic Review and Meta-Analysis. Scientific Reports, 2018. 8(1): p. 12050.

3. Biron, B.M., A. Ayala, and J.L. Lomas-Neira, Biomarkers for Sepsis: What Is and What Might Be? Biomarker insights, 2015. 10(Suppl 4): p. 7-17.

4. Huang, K.-J., et al., An interferon-γ-related cytokine storm in SARS patients. Journal of Medical Virology, 2005. 75(2): p. 185-194.

5. Cao, Y., et al., Global transcriptome analysis of H5N1 influenza virus-infected human cells. Hereditas, 2019. 156: p. 10-10.