使用雷射誘導石墨烯電極於細胞基底行為電阻抗感測

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姓名

江易宸(YI-CHEN JIANG) 查詢紙本館藏

畢業系所

電機工程學系

論文名稱

使用雷射誘導石墨烯電極於細胞基底行為電阻抗感測
(Laser-Induced Graphene (LIG) Electrode for Electric Cell–Substrate Impedance Sensing)

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摘要(中)

本實驗利用電化學交流阻抗法即時監測動物細胞貼附於電極上的行為，這項技術被稱為Electric Cell–substrate Impedance Sensing (ECIS)。檢測的項目為不同生長情況的小鼠纖維母細胞（L929）於電極上因懸浮或貼附所引起的阻抗值變化。本研究所使用的感測元件是利用雷射刻劃在耐高溫、耐化學藥品的聚醯亞胺(Polyimide)基板表面所製作出的石墨烯電極，稱為雷射誘導石墨烯(Laser-induced graphene, LIG)電極。電極型態上我們將選擇指叉電極；經過許多不同的電極尺寸、電極數量的設計與量測結果，最終將選擇考慮細胞大小以及LIG所能達到的尺寸之條件下期望得到穩定且靈敏度高的電極之尺寸。將小鼠纖維母細胞(L929)放置在指叉電極表面上的培養液中生長，使用低真空掃描式電子顯微鏡(LV-SEM)讓我們能夠清楚觀察在電極上面的增殖、生長、貼附的情形。監測上，利用電化學阻抗分析儀量L929細胞在含有不同濃度的毒化物或是藥品之培養液中生長時細胞和基底電極間的阻抗頻譜隨時間之變化。實驗結果顯示毒化物對細胞生長、貼附行為的影響可以利用測量LIG電極的阻抗頻譜而即時監控。本方法的優點包括可靠性高、分析速度快、精確性高，而且使用的樣品數和試劑少，就可以獲得相當好的樣品資訊。

摘要(英)

This study used electrochemical AC (alternating-current) impedance method to monitor the behavior of animal cells attached to electrodes in real time, i.e., to conduct Electric Cell-substrate Impedance Sensing (ECIS). The target of detection was the impedance changes caused by suspension or attachment of mouse fibroblasts (L929) in different growth conditions on the interdigital electrodes. Laser-induced graphene (LIG) electrodes were used for measuring the cell-substrate impedance as well as for the substrate on which cells attached and grew. They were fabricated by laser scribing on the surface of heat-resistant, chemical-resistant polyimide film. Mouse fibroblasts (L929) were grown in culture medium on the surface of the interdigitated electrodes. Low-vaccume scanning electron microscopy (LV-SEM) allowed us to visually verify the cell proliferation, growth, and attachment on the electrodes. Then, the impedance spectrum between the cells and the substrate electrode was measured with time when L929 cells were grown in culture media containing different concentrations of poisons or drugs by electrochemical impedance analyzer. The experimental results showed that the feasibility of real-time monitoring the effects of poisons on cell growth and attachment behavior by measuring the cell-substrate impedance spectrum with LIG electrodes. The advantages of this method include high reliability, high analysis speed, high accuracy, and relatively good sample information with a small number of samples and reagents used.

關鍵字(中)

★ 細胞基底行為電阻抗感測
★ 雷射誘導石墨烯電極
★ 指叉電極
★ 阻抗式生物感測器
★ 細胞貼附
★ 毒化物檢測

關鍵字(英)

★ ECIS
★ laser-induced graphene electrode
★ interdigital electrode
★ impeditive biosensor
★ cell attachment
★ toxic detection

論文目次

中文摘要 i
Abstract ii
致謝 iii
目錄 iv
圖目錄 vii
表目錄 xiii
第一章緒論 1
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 交流阻抗法電化學生物感測器測量原理 2
1-1-4 指叉型電極原理介紹 5
1-2 細胞生理特性介紹 8
1-2-1 細胞結構 8
1-2-2 細胞的主要成分 12
1-2-3 分子穿越細胞膜之移動方式 14
1-2-4 細胞生長週期 15
1-2-5 細胞的貼附 16
1-2-6 細胞凋亡與衰老 17
第二章研究動機與目標 19
2-1 研究動機 19
2-2 研究目標 19
第三章實驗方法 20
3-1 阻抗感測元件設計 20
3-2 雷射雕刻機基本設備介紹以及軟體的使用 22
3-2-1 雷射雕刻機之感測晶片材料選用 22
3-2-2 儀器設備 23
3-2-3 製作電極 UCP 軟體之基本操作介面以及相關參數設定 24
3-3 細胞培養 29
3-3-1 細胞培養藥劑、材料 29
3-3-2 細胞培養使用之實驗設備 32
3-3-3 細胞培養方法及步驟 37
3-3-4 細胞用LV-SEM顯微鏡觀察 40
3-4 小鼠纖維母細胞（L929）阻抗量測 42
3-4-1 小鼠纖維母細胞（L929）阻抗量測方法實驗 42
3-4-2 IM6-ex儀器量測阻抗實驗 48
3-4-3 IM6-ex儀器初始設定 50
3-4-4 等效電路設計 52
第四章實驗結果與討論 54
4-1 自製指叉電極之拉曼光譜 54
4-2電極阻抗、電容值量測結果 56
4-2-1 電極電容值特性 56
4-2-2 溶液下等效電路探討 60
4-2-3 不同溶液阻抗電容值量測以及擬合結果 61
4-3 小鼠纖維母細胞（L929）阻抗量測實驗 65
4-3-1 小鼠纖維母細胞（L929）生長情況 65
4-3-2 小鼠纖維母細胞（L929）貼附電極之等效電路探討 69
4-3-3 小鼠纖維母細胞（L929）貼附電極之阻抗值量測 70
4-3-4 小鼠纖維母細胞（L929）添加Trypsin–EDTA之阻抗值量測 76
4-3-5 小鼠纖維母細胞（L929）添加 DMSO 之阻抗值量測 85
4-4 小鼠纖維母細胞（L929）貼附電極之 SEM 顯微鏡圖 97
第五章結論 102
參考文獻 103

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指導教授

蔡章仁(Jang-Zern Tsai)

審核日期

2022-8-2

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