設計一網印電極藉由電阻抗感測偵察細胞行為

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姓名

林盈瑞(Ying-Jui Lin) 查詢紙本館藏

畢業系所

電機工程學系

論文名稱

設計一網印電極藉由電阻抗感測偵察細胞行為
(Screen-printed electrode design for cell behaviors detection by electric cell-substrate impedance sensing)

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摘要(中)

本研究設計一拋棄式銀電極作為量測工具，以細胞與基底間電性檢測法(ECIS)為分析方法，針對老鼠纖維母細胞L929進行即時監控，以及小鼠巨噬細胞RAW264.7發炎行為來驗證電極設計原理。指叉電極由網印方式製造， LCR電表確認其基本電氣特性。在細胞阻抗實驗量測中，自製指叉電極能準確監控細胞L929在電極表面之行為，當細胞貼附在一對電極表面狀態十二小時下中頻歸一阻抗變化(NIC)最高可達800%。實驗過程中搭配掃描式電子顯微鏡技術(SEM)驗證細胞型態與阻抗之關係。為了驗證電極設計原理，本研究利用小鼠巨噬細胞進行發炎反應，當電極尺寸貼近細胞尺寸狀態時可有效提升感測器整體之阻抗靈敏度。上述實驗證明出當細胞貼附在電極表面時，電極對數為一對，而間距以及寬度為50μm之電極規格具有最高阻抗靈敏度，可減少電極表面之電場穿越長度使電場強度侷限在電極表面。除此之外為了確認生物相容性此網印電極利用MTT分析方法來驗證此電極適合細胞貼附。

摘要(英)

In this study, aimed to monitor L929 cell behaviors in real time, a disposable silver electrode was designed and incorporated in an electric cell-substrate impedance sensing analysis. Inflammation of cell RAW264.7 was developed to verify the electrode design theory. The electric characteristic of the electrode was measured with an LCR meter. This home-made interdigitated electrode was fabricated by screen printing technique. In the cell impedance measurement experiment, the normalized impedance change (NIC) of cell attaching on the one-paired electrode could be 800%, revealing the potentiality of our home-made electrodes to monitor cell behaviors on the electrode. The relationship between the measured impedance and the cell morphology on the electrode would be explained by scanning electron microscope (SEM) technique. Inflammation of cell RAW264.7 was developed to verify the electrode design theory. When the cell size was commensurate with the electrode size, the sensitivity of our sensor would be increased. Our experiment results showed that the one-paired electrode, with 50 μm spacing and width, had the highest sensitivity, supposedly because of more concentrated electric field distribution surfacing the substrate and electrode. Besides, MTT assays verified the compatibility of our home-made electrodes.

關鍵字(中)

★ 指叉電極
★ 細胞與基底間電性檢測法
★ 掃描式電子顯微鏡
★ 靈敏度

關鍵字(英)

★ interdigitated electrode
★ electric cell-substrate impedance sensing
★ scanning electron microscope
★ sensitivity

論文目次

中文摘要……………………….………………………………………………..……..І
Abstract……………………………………………………………………….….……II
致謝…………………………………………………………………………….….…III
目錄……………………………………………………………………….………….IV
圖目錄.......……………………………………………………………………...….VIII
表目錄……………………………………………………………..…………..…XXIII
第一章諸論
1-1 前言…………………………………………………………………………….....1
1-1-1 生物感測器定義………………………………………………………1
1-1-2 生物感測器組成與結構………………………………………………1
1-1-3 電化學生物感測器量測方法…………………………………………4
1-1-4 網印技術應用在生物感測器…………………………………………8
1-2 細胞生理特性介紹……………………..…………………………….…………10
1-2-1 細胞結構……………………………………………………………..10
1-2-2 分子穿越細胞膜的移動……………………………………………..13
1-2-3 細胞生長週期………………………………..………………………14
1-3 文獻回顧………………………………………………..……………………….15
1-3-1 阻抗量測應用在細胞反應……………………………………………...15
1-3-1-1 細胞貼附…………………………………………………………16
1-3-1-2 細胞成長…………………………………………………………26
1-3-1-3 細胞凋亡…………………………………………………………30
1-3-1-4 細胞分化…………………………………………………………37
1-3-2 細胞阻抗量測之電極設計……………………………………………...43
1-3-2-1 指叉電極幾何參數之設計…….……………………….………..45
1-3-2-1-1 指叉電極寬度跟間距影響…………………….…………47
1-3-2-1-2 指叉電極長度與對數影響………………….……………58
1-3-2-1-3 指叉電極厚度影響………….……………………………62
第二章研究動機與目標………………………….……………………………….66
2-1 研究動機. …………..…….…………………….……………………………….66
2-2 研究目標……………………………….……….……………………………….66
第三章實驗方法…………….…………………....……………………………….67
3-1 電極製備與電性分析…………………………………………………………..67
3-1-1 電極製備之儀器以及材料………………………………….…………..67
3-1-2 電極電容值分析……….………………………………………….…….74
3-1-3 不同溶液阻抗量測…………………………………………………..….76
3-1-4 電極在細胞培養溶液下之阻抗量測………………….……..…………77
3-2 細胞培養……………………………………………………………..………….78
3-2-1 細胞培養藥劑、材料及使用設備…………………………..………….78
3-2-1-1 細胞培養藥劑以及材料……………………………..…………..78
3-2-1-2 細胞培養實驗設備…………………………………..…………..79
3-2-2細胞培養方法及步驟……………………………………………………83
3-2-2-1 細胞解凍以及培養………………………………………………84
3-2-2-2 細胞分盤以及繼代培養…………………………………………84
3-2-2-3 細胞計數…………………………………………………………85
3-2-2-4 細胞冷凍…………………………………………………………86
3-3 鼠纖維母細胞(L929)細胞阻抗量測………………………………….………..86
3-3-1 指叉電極之細胞阻抗48hr量測……………………………..………….86
3-3-2 電極不同對數之阻抗量測………………………………….…………..90
3-3-3 電極不同對數之靈敏度量測………………………………….………..91
3-3-4 電極與細胞等效模型量測………………………………….…………..91
3-3-5 細胞行為與掃描式電子顯微鏡應用…………………………….……..93
3-3-6 電極與細胞活性實驗……………………………………………...……96
3-4 小鼠巨噬細胞(RAW264.7)阻抗量測與電極設計原理驗證……………...…100
3-4-1 指叉電極微小之原理驗證與實驗…………………………...……..…100
第四章實驗結果…………………………………………………………………108
4-1 電極阻抗量測與製程評估結果………………………………………………108 4-1-1電極製程結果分析…………………………………..…………………108
4-1-2 電極電性分析…………………………………….……..………..……120
4-1-3 不同溶液阻抗分析……………………………………….……………121
4-1-4 電極在細胞培養溶液下阻抗分析………………….…………………144
4-2 鼠纖維母細胞(L929)阻抗量測結果…………………….……………………148
4-2-1 指叉電極之細胞阻抗48hr量測………………………………………154
4-2-2 電極不同對數之阻抗分析…………………………………………….157
4-2-3 電極不同對數之靈敏度分析………………………………………….164
4-2-4電極與細胞等效模型綜合分析………………………………………..168
4-2-5 細胞行為與電極阻抗關係…………………………………………….186
4-2-6電極與細胞活性分析…………………………..………………….……192
4-3 小鼠巨噬細胞(RAW264.7)阻抗量測結果…………….………………….…..196
4-3-1 控制組阻抗紀錄與模擬結果………………..……………………..196
4-3-2 實驗組阻抗紀錄與模擬結果…………..…………………………..205
4-3-3 靈敏度比較與原理驗證…………….……….……………………..215
第五章結論………………………………………………….………….………..221
未來展望……………………………………………………………………………222
參考文獻…………………………………………………………………………....223

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指導教授

蔡章仁(Jang-Zern Tsai)

審核日期

2013-1-31

推文