近幾十年來，全球陷入癌症的恐慌之中，雖然現今科技已經發展出許多針對惡性腫瘤的醫療技術，但是癌症的可怕之處不在於腫瘤細胞，而是在其散佈全身的轉移性，我們稱此細胞為循環腫瘤細胞(CTCs)。癌症的轉移不只讓藥物治療的效果降低，更增添癌症治癒的困難度，故利用氧化石墨烯材料進行抓取細胞並使用高靈敏度的電訊號偵測，以達到循環腫瘤細胞的快速篩檢功效，使人類及早發現且加以治療，有效降低癌症的高死亡風險。
在此篇研究中，我們製備氧化石墨烯(GO)材料，以氧化石墨烯表面官能基跟溴化十六烷基三甲銨(CATB)正負電相吸形成模板，利用雙層乳化法將乳化層包覆在模板內，形成有機相包覆水相之二氧化矽球，接者利用高溫絕氧鍛燒方式，將模板去除並形成中空的二氧化矽孔洞球(PSG)。使用掃描式及穿透式電子顯微鏡(SEM與TEM)可以鑑定其為吸附在氧化石墨烯表面的球狀結構。
氧化石墨烯表面的樹枝狀粗糙結構可以有效的增加與細胞的接觸面積並促進細胞的貼附，有別於氧化石墨烯平面的粗糙度變化，二氧化矽球除了增加表面粗糙度，也可以構築出空間的高低落差，我們稱此為三維粗糙度的改變，以達到更高的細胞抓取效率。此外，為了達到檢測血液中癌細胞的目標，將西妥昔單抗以化學鍵結的方式鍵結在材料表面用以增加材料的標靶特性，並利用紫外可見光譜以及原子力顯微鏡鑑定。抓取細胞的定量檢測則可以藉由電化學感測的高靈敏且快速應答等優點，利用循環伏安法與交流阻抗法來探測細胞黏附在電極表面時電解質阻抗之變化。經由氧化石墨烯表面粗糙度的改變以及修飾上二氧化矽球的三維粗糙度變化可以有效的增加細胞的貼附，並利用即時且靈敏的電化學檢測將細胞定量，此研究的短時間抓取以及檢測適合應用於血液的快速篩檢平台，並期許未來能夠在循環腫瘤細胞的血液檢測有更多方面性應用。

Cancer has occupied the symbol of death for years. The devastating effect of this complex illness is due to the progression from a localized to metastatic disease. We call it circulating tumor cells (CTCs). Early detection of cancer cells is important for clinical diagnostics, toxicity monitoring, and public health protection.
Graphene materials illustrated to be a biocompatible material nowadays are commonly used in the CTCs capture and detection due to the surface roughness structure which is believed to enhance the surface area-to volume ratio with cells. In this work, we construct carbon nanospheres coated on graphene oxide (PSG) not only to increase surface roughness but for the difference in height called 3D roughness difference. The capture efficiency indicates that the 3D roughness does effect the cell adhesions compared with graphene oxide and smooth ITO glass.
Rapid detection of blood includes both detection and specificity. Erbitux is conjugated with materials by the peptide bond and illustrated by UV-VIS and atomic force microscopy. Detection of captured cells by electrochemical detection can provide high sensitivity and rapid response. Here, cyclic voltammetry (CV) and AC impedance are used for device examination and cell quantification.
This detecting method adapting roughness difference to enhance cell adhesion and quickly response by electrochemical detection can accurately quantify the captured cells on the surface which is suitable for CTCs examination or other chemical detection platform.

中文摘要 I
Abstract II
致謝 III
Table of Contents V
List of Schemes VII
List of Figures VIII
1.1 Circulating Tumor Cells 1
1.2 Materials and Cell Capture 2
1.2.1 Techniques for CTC Isolation 2
1.2.1.1 Affinity-based Isolation 2
1.2.1.2 Size-Based Separation of CTCs 3
1.2.1.3 Dielectric Separation of CTCs 4
1.2.2 After Isolation, Cell Capture and Analysis 5
1.2.2.1 Marker-Free Isolation of CTCs 5
1.2.2.2 Capture and Release of CTCs 6
1.2.2.3 Enabling Culture and Expansion 8
1.2.3 Clinical Technology 8
1.2.3.1 Line-confocal Microscope Detection 9
1.2.3.2 Surface-Enhanced Raman Scattering Detection 10
1.2.3.3 CTCs Detection with Negative Enrichment 12
1.2.3.4 Positive Enrichment – In vivo Measurement 12
1.2.3.5 Positive Enrichment – In vitro Measurement 14
1.3 Graphene for CTC Analysis 17
1.4 Electric Signal and Quantization 20
Chapter 2 Experimental Section 23
2.1 Chemicals 23
2.2 Apparatus 23
2.3 Methods 24
2.3.1 Synthesis of Graphene Oxide 24
2.3.2 Synthesis of Porous Silica Nanospheres @ GO (PSG) 25
2.3.3 Preparation of GO/PSG@PEDOT Chip (GPC/SPC) 25
2.3.4 Erbitux Immobilization 26
2.3.5 Characterization 27
2.3.6 Cell Culture 27
2.3.7 Cell Capture and Sample Produce for SEM Image 28
2.3.8 Cell Staining 28
2.3.9 Cytotoxicity of ITO Chips 29
2.3.10 Electrochemical Measurement 29
Chapter 3 Results and Discussion 30
3.1 Electric Detection of CTCs by GO and PSG 30
3.2 Synthesis and Characterize of GO and PSG 30
3.3 Cell Adhesion on GO and PSG 34
3.3.1 Roughness of Materials 35
3.3.2 Capture Efficiency by Flow Cytometer 36
3.3.3 Image of Captured Cells 38
3.4 Erbitux Immobilization 40
3.5 CTCs Detecting Chips 42
3.5.1 Cell Viability of ITO Chips 43
3.5.2 GO/PSG@PEDOT Chip (GPC/SPC) 43
3.5.3 Cyclic voltammetry (CV) for GPC/SPC 44
3.5.4 Erbitux conjugation on GPC/SPC 45
3.6 Captured cells detecting by CV and EIS 46
3.7 Macrophage Test 48
Chapter 4 Conclusion 49
Reference 50

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