老鼠纖維母細胞L929貼附於微電極所造成的阻抗變化之模擬

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研究生：	黃翊軒 Yi-Shiuan Huang
論文名稱：	老鼠纖維母細胞L929貼附於微電極所造成的阻抗變化之模擬 Simulation of mouse fibroblasts L929 attached on micro scale electrodes caused of impedance change
指導教授：	蔡章仁
口試委員:
學位類別：	碩士 Master
系所名稱：	資訊電機學院 - 電機工程學系 Department of Electrical Engineering
論文出版年：	2014
畢業學年度：	102
語文別：	中文
論文頁數：	226
中文關鍵詞：	阻抗式生物感測器、有限元素模擬、細胞貼附
外文關鍵詞：	Impedance sensor, FEM simulation, cell adhesion
相關次數：	點閱：7 下載：0
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研究細胞的貼附能夠幫助我們了解細胞的生長行為，對於利用活體細胞測試毒物或是藥品對細胞活性的影響，細胞貼附與否是判斷細胞生長情形的重要指標之一。近來量測細胞的電阻抗特性逐漸取代了細胞染色法，藉由分析細胞的阻抗特性，能夠幫助我們迅速且有效觀察細胞特徵行為。本研究提供一研究細胞貼附對阻抗影響的新方法，根據L929細胞貼附過程中細胞形態的變化，分別建構6個模型，模擬討論細胞貼附過程中，量測阻抗因電極上受細胞與細胞外基質蛋白(ECM)覆蓋的面積不同、細胞-電極距離的不同，所造成的阻抗變化特性。研究結果顯示，隨著電極表面受細胞覆蓋的面積愈大、細胞-電極距離愈小，會使得中頻的歸一阻抗變化量(NIC)愈大；隨著電極表面受細胞外基質(ECM)覆蓋的面積愈大，會使得低頻的歸一阻抗變化量(NIC)愈小，當電極表面受細胞與ECM覆蓋面積達92.16 %時，中頻(100 kHz)阻抗變化值達1260 %，低頻(1 Hz)阻抗變化達-81.9 %。模擬結果說明了當量測電極尺寸與表面所覆蓋的細胞面積愈相近時，細胞貼附所造成的阻抗變化會愈明顯，也說明了細胞貼附時所分泌產生的ECM物質，會造成低頻阻抗量測值下降的現象。

Analysis of cell adhesion property can help us to understand cell growth situation and test cytotoxicity effect of different toxicants to cells. Recently, electrical cell substrate impedance sensing (ECIS) replaced cell staining; analyzing the impedance characteristics of cells will help us to study the situation and property of cell growth behavior rapidly and effectively. In this study, we proposed a new approach to analyse the L929 cell growth on electrode surface using finite element method (FEM) simulation. According to L929 cell morphology change on cell covered electrode surface during growth, six electrical models were designed respectively. FEM simulation was carried out to measure impedance response due to the change of coverage cell, extracellular matrix (ECM) area on electrode, and the change of resistive gap region of cell-electrode. The results showed that the larger cell covered area on electrode and shorter cell-electrode gap caused maximμm change in normalized impedance change (NIC) at intermediate frequency (100 kHz), and at lower frequencies (1 Hz) minor change in normalized impedance change (NIC) was observed due to ECM covered on electrode surface. When the coverage of cell and ECM reached 92% of total electrode area, the impedance at intermediate frequency changed by 8 folds and at lower frequency impedance changed by -0.8 folds from that of measured impedance in bare electrodes. FEM simulation results showed that when cell covered area is similar to electrode size, the significant change in impedance will be observed. The result also indicated the ECM protein produced by cell attachment will contribute to decrease the impedance at lower frequency.

摘要    VI
ABSTRACT    VII
致謝    IX
目錄    X
圖目錄    XV
表目錄    XXI
第一章    緒論    1
1.    前言    1
1.1.    生物感測器定義    1
1.2.    生物感測器的組成與結構    2
1.3.    交流阻抗法電化學生物感測器量測方法    3
2.    細胞結構及其生長與貼附    6
2.1.    細胞結構    6
2.2.    細胞的生長    9
2.3.    細胞的貼附    10
2.3.1.    外部生成物的聯結與功能    12
2.3.2.    細胞外基質蛋白(ECM)    13
2.3.3.    細胞貼附的過程    16
第二章    文獻回顧    19
1.    細胞貼附過程的細胞形態    19
2.    電極阻抗量測特性模擬    23
2.1.    電極厚度對阻抗的重要性與影響    24
2.2.    電極表面電雙層對阻抗的重要性與影響    26
2.3.    電極在溶液中量測的等效電路模型    28
3.    細胞阻抗量測特性模擬    32
3.1.    細胞懸浮模型    32
3.2.    細胞貼附模型    41
3.3.    細胞貼附的等效電路模型    60
4.    電極表面ECM對阻抗的影響    68
4.1.    細胞阻抗量測時低頻阻抗下降的現象    68
4.2.    ECM與細胞覆蓋於電極表面的等效電路模型    74
4.2.1.    電極表面進行蛋白質改質的等效電路模型    74
4.2.2.    考慮ECM影響的細胞貼附模型：    76
5.    模擬結果的分析    81
5.1.    模型的Y-Z切面電位分析    81
5.2.    模型不同Z方向高度下的切面電位分析    84
5.3.    系統的阻抗分析    86
5.4.    系統的歸一阻抗變化(NIC)分析    86
第三章    研究動機與目標    88
1.    實驗動機    88
2.    實驗目標    89
第四章    實驗方法    90
1.    軟體與模組    90
2.    模型幾何結構設計    91
2.1.    模擬時設計電極厚度與否之比較    91
2.2.    細胞隨時間貼附於平行電極之模擬    92
2.3.    細胞隨時間貼附電極表面ECM影響之模擬    95
3.    材料參數給定    99
3.1.    模擬時設計電極厚度與否之比較    99
3.2.    細胞隨時間貼附於平行電極之模擬    101
3.3.    細胞隨時間貼附電極表面ECM影響之模擬    103
4.    邊界條件設定    105
5.    網格劃分與求解    107
6.    分析方法與後處理    108
6.1.    切面電位分析    108
6.1.1.    Y-Z切面電位分析    109
6.1.2.    不同Z方向切面電位分析    110
6.2.    系統阻抗分析    112
6.2.1.    系統總阻抗分析    112
6.2.2.    系統的歸一阻抗變化值(NIC)分析    113
6.3.    系統相位分析    113
第五章    實驗結果與討論    115
1.    模擬時設計電極厚度與否之比較    115
1.1.    電極模型的系統阻抗分析    116
1.2.    模型的Y-Z切面與不同Z方向切面電位分析    123
1.2.1.    低頻100 HZ    123
1.2.2.    中頻100 KHZ    128
1.2.3.    高頻100 MHZ    131
1.3.    電極有厚度模型在DI WATER中的阻抗分析    133
1.4.    結論    134
2.    細胞隨時間貼附於平行電極之模擬    136
2.1.    系統的阻抗分析    139
2.2.    模型的歸一阻抗變化(NIC)分析    147
2.3.    模型的Y-Z切面與模型不同Z方向切面電位分析    151
2.3.1.    低頻100 HZ    151
2.3.2.    中頻100 KHZ    155
2.3.3.    高頻100 MHZ    160
2.4.    結論    163
3.    細胞隨時間貼附電極表面ECM影響之模擬    165
3.1.    模型的系統阻抗分析    166
3.2.    模型的歸一阻抗變化(NIC)分析    176
3.3.    模型的Y-Z切面與模型不同Z方向切面電位分析    181
3.3.1.    低頻100 HZ    181
3.3.2.    中頻100 KHZ    185
3.3.3.    高頻100 MHZ    189
3.4.    結論    193
第六章    結論    194
參考文獻    197

                                

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