指叉電極檢測系統以阻抗分析方式檢測老鼠纖維組織細胞L929的新陳代謝及增殖

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姓名

蔡孟廷(Meng-ting Tsai) 查詢紙本館藏

畢業系所

電機工程學系

論文名稱

指叉電極檢測系統以阻抗分析方式檢測老鼠纖維組織細胞L929的新陳代謝及增殖
(Impedance analysis of metabolism and proliferation of mouse fibroblast cells L929 by a portable interdigitated microelectrodes detection system)

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摘要(中)

本研究利用指叉型電極來檢測老鼠纖維母細胞L929生長代謝及分裂後時阻抗的變化。指叉型電極利用微影製程的方式製作，電極尺寸為500um x 8um x 50nm。本研究將L929附著於生物感測晶片上，然後量測不同濃度及不同生長情況下阻抗變化情況。在進行阻抗量測前，會先於感測晶片表面上一層Poly-L-lysine使細胞更容易貼附。初步結果為不同細胞數量相對造成不同的阻抗變化，其原因為細胞代謝物會影響電雙層電容的變化。為了達到方便及快速檢測，本研究利用微控制器、生物感測晶片及阻抗量測元件 AD5933來發展簡易的阻抗量測系統。利用此阻抗量測系統初步可以觀察到在不同細胞濃度下其代謝對阻抗造成的變化。因此，此生物檢測系統利用阻抗量測方式確實對於細胞代謝的量測提供一個有效的分析方式。

摘要(英)

Interdigitated microelectrodes were used to monitor the growth of L929 cells during metabolism and proliferation. The interdigitated microelectrodes were fabricated with microfabrication technology with 8μm of width, 8μm of gap, and 500μm of length for each electrode. In this study, the mouse fibroblast cells L929 was cultured on the electrodes surface with pre-coated poly-L-lysine. The impedance change in the interdigitated electrode pair was measured during cells adhesion, spreading, and proliferation. The experimental results show that cells concentration increase could affect the impedance change because the cell’s metabolite correlated to the double layer capacitance between the electrodes and the cells. Further, to develop a portable and fast detection device, we built an impedance detection system using a microprocessor, the impedance convertor AD5933, and the interdigitated microelectrodes chip. Using this impedance detection system can prove to observe metabolism of different concentration for cells growth stage. Thus, this cell-growth monitor system by impedance measurement provides a useful analytical method for cells metabolism.

關鍵字(中)

★ 阻抗量測
★ 細胞培養
★ 電雙層
★ 指叉電極

關鍵字(英)

★ double layer
★ cell culture
★ impedance measurement
★ interdigitated electrode

論文目次

中文摘要 I
英文摘要 II
誌謝 III
目錄 IV
表目錄 XIV
第一章緒論 1
1-1 前言 1
1-2 生物感測器介紹 2
1-2-1 歷史發展 2
1-2-2 定義及構成 3
1-2-3 類型及偵測原理 4
1-3交流阻抗分析背景 10
1-4 阻抗感測器的量測特性 12
1-5 指叉型電極介紹 13
1-6 細胞生理特性介紹 16
1-6-1細胞結構 16
1-6-2 分子穿越細胞膜的移動 19
1-6-3 細胞生長周期 22
1-6-4細胞衰老與凋亡 23
第二章研究背景 25
2-1 研究動機 25
2-2 研究目標 26
第三章阻抗式感測器製作及實驗方法 28
3-1 阻抗式感測晶片製作 28
3-1-1感測晶片設計 28
3-1-2感測晶片材料選用 31
3-1-3實驗材料及儀器設備 31
3-1-3-1 實驗材料 31
3-1-3-2 儀器設備 31
3-1-4 製程步驟與相關參數 34
3-2 細胞培養 39
3-2-1 細胞培養藥劑、材料及使用設備 40
3-2-1-1 細胞培養藥劑及材料 40
3-2-1-2 實驗設備 40
3-2-2 培養方法及步驟 43
3-2-3 細胞計數方法 44
3-3 細胞阻抗量測實驗 45
3-3-1 培養方法及步驟 47
3-3-2 IM6-ex量測實驗 48
3-3-2-1 電路量測方式及相關參數 48
3-3-2-2 等效電路設計 49
3-3-3 AD5933 量測實驗 53
3-3-3-1 電路量測原理 53
3-3-3-2 AD5933 Demo board量測程式操作說明 59
3-3-3-3 自製電路設計及量測原理 60
3-3-3-4 量測程式設計 65
第四章結果與討論 67
4-1 製程結果分析 67
4-2 晶片品質及特性分析 70
4-2-1 電容特性 70
4-2-2 交流阻抗特性 72
4-2-2-1 溶液下等效電路探討 72
4-2-2-2 不同溶液下阻抗分析 74
4-2-2-3 等效電路fitting結果 77
4-3 阻抗式量測電路整合結果 78
4-3-1量測電路實現 78
4-3-2 PC端程式測試 80
4-4 細胞阻抗量測結果 82
4-4-1 細胞吸附情況 82
4-4-2 IM6-ex量測結果 83
4-4-2-1 細胞生長曲線阻抗量測結果 85
4-4-2-2 擬合阻抗量測結果 92
4-4-2-3 細胞濃度阻抗量測分析 96
4-4-2-4 細胞活性及代謝測試 100
4-4-3 AD5933 Evaluation Board量測結果 100
4-4-3-1 細胞生長曲線阻抗量測結果 102
4-4-3-2 細胞濃度阻抗量測分析 104
4-4-4 自製電路量測結果 105
4-4-4-1 細胞生長曲線阻抗量測結果 107
4-4-4-2 細胞濃度阻抗量測分析 108
第五章結論 110
未來展望 113
參考文獻 114

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指導教授

蔡章仁(Jang-zern Tsai)

審核日期

2009-8-26

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