開發生物阻抗量測系統搭配網印指叉電極採用電化學阻抗頻譜法偵測人類白蛋白與細胞行為

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姓名

王冠堯(Kuan-Yao Wang) 查詢紙本館藏

畢業系所

電機工程學系

論文名稱

開發生物阻抗量測系統搭配網印指叉電極採用電化學阻抗頻譜法偵測人類白蛋白與細胞行為
(Development of bio-impedance measurement system to detect human serum albumin and cell behavior using screen-printed interdigitated electrodes by electrochemical impedance spectroscopy)

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摘要(中)

交流阻抗法為一非侵入式檢測技術，其可藉由細胞、組織與器官的電特性及其變化規律提取生理、病理狀況相關的生物醫學信號。本研究利用電極的尺寸維度接近小鼠巨噬細胞發炎後的大小，並藉由交流阻抗法即時監測小鼠巨噬細胞發炎的情形，搭配掃描式電子顯微鏡來觀測小鼠巨噬細胞於電極上發炎的狀況，並探討所量出之阻抗值與細胞發炎之間的關係。本研究也探討細胞貼附於電極時對於低頻阻抗值所造成之影響，在此我們檢測蛋白質濃度，以探討蛋白質濃度與低頻阻抗值之間的關係。小鼠細胞發炎的實驗中，其當細胞發炎之實驗組會導致中頻阻抗值變化上升210%，而細胞沒有發炎之控制組之中頻阻抗變化僅上升80%，成功利用交流阻抗法去監測細胞發炎的情形。在量測細胞低頻的阻抗實驗中，其低頻的阻抗值與細胞外基質濃度具有其線性關係，低頻組抗值分析出細胞貼附，而中頻阻抗值分析細胞行為，結合其兩種分析可以得知完整的細胞行為。除此之外，本研究也開發自製阻抗量測系統，其利用微控制器(AT89S51)和積體電路(AD5933)以達到便宜、使用方便、容易攜帶及高靈敏度的阻抗量測，並量測人類白蛋白進行測試，利用所開發的阻抗量測系統量測之阻抗值與精密阻抗量測儀器Im6/6ex相近，其差異僅6%以下。

摘要(英)

Bio-impedance technology extracts both physiological and pathological information from the human body through the use of biological cells, tissues, and organs. This form of testing is a non-destructive technique, which is widely in use. In our study, we used screen-printed interdigitated electrodes to detect inflammation of RAW264.7 cells in real time. We use the scanning electron microscope (SEM) to observe the cell morphology on the electrode. The relationship between impedance changed and inflammation was observed. This study also research the effect of the cells attached to the electrode at low frequencies. We will do protein quantitative analysis which will observe the relationship with impedance. In inflammation of RAW264.7 experiment for 24 hour, the inflammation groups increased normal impedance changed to 210%, while the control groups just increased normal impedance changed to 80%. In study on electrical properties of electrode surfacing with culture L929 at low frequency experiment, the results show the relationship between concentration of protein and impedance at low frequency. This study also development the impedance measurement system. The system we designed consists of two components. The first component is an AT89S51 microcontroller. The second is the integrated circuit AD5933, a bio-impedance measuring device manufactured by Analog Devices. Working together, the AT89S51 and AD5933 combine to produce a portable bio-impedance detection system. In human serum albumin experiment, the impedance measured by homemade impedance detection systsm and Im6/6ex are similar. The difference between homemade detection system and Im6/6ex are 6% or less.

關鍵字(中)

★ 生物感測電極
★ 人類白蛋白
★ 細胞發炎

關鍵字(英)

★ Bioelectrode
★ Inflammation

論文目次

中文摘要 I
Abstract II
致謝 IV
目錄 V
圖目錄 VIII
表目錄 XVII
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 電化學生物感測器量測方法 2
1-1-4 電極應用於生物感測器 21
1-2 生物臨床檢測 25
1-2-1蛋白尿檢測 25
1-2-2 細胞檢測 27
第二章研究動機與目標 70
2-1 研究動機 70
2-2 研究目標 70
第三章實驗方法 72
3-1 細胞培養 72
3-1-1 細胞培養藥劑、材料及使用設備 72
3-1-2 細胞培養方法及步驟 79
3-1-3 細胞SEM處理 82
3-2 細胞阻抗量測驗 84
3-2-1 小鼠巨噬細胞(RAW264.7)阻抗量測實驗 84
3-2-2 L929細胞於低頻阻抗量測與蛋白質檢測濃度 88
3-3 阻抗感測系統設計 94
3-3-1阻抗感測系統設計原理 94
3-3-2 AD5933內部架構 95
3-3-3 AT89S52 微控制器軟體設計 100
3-3-4 LabVIEW人機介面介紹 108
3-3-5 整體電路實現 112
3-4 人類白蛋白阻抗量測 114
3-4-1 人類白蛋白於不同背景溶液下阻抗量測實驗 114
3-4-2 臨床尿液人類白蛋白阻抗量測實驗 115
第四章實驗結果 117
4-1小鼠巨噬細胞(RAW264.7)阻抗量測 117
4-2 L929細胞於低頻阻抗量測與蛋白質檢測濃度 135
4-3 阻抗量測系統測試及改良 137
4-3-1 阻抗量測系統之測試結果 137
4-3-2 阻抗量測系統之改良 140
4-3-3 人類白蛋白阻抗量測實驗結果 148
第五章結論 156
Reference 157

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指導教授

蔡章仁(Jang-Zern Tsai)

審核日期

2013-8-30

推文