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研究生： 張明軒
Chang, Ming-Hsuan
論文名稱： 藉由隙環共振器顯微技術拍攝在近紅外之活體細胞內部影像
Intracellular Imaging of Living Cells by Split Ring Resonators Microscopy in Near-Infrared (NIR) Region
指導教授： 嚴大任
口試委員: 嚴大任
張雍
王子威
學位類別： 碩士
Master
系所名稱： 工學院 - 材料科學工程學系
Materials Science and Engineering
論文出版年： 2013
畢業學年度： 101
語文別： 英文
論文頁數： 76
中文關鍵詞： 超材料 、活體細胞 、內部影像 、近紅外
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    • 超材料為人工電磁材料，是由單位尺寸小於入射光波長所組成的結構。超材料的性質來自於內部結構產生共振的集合響應，所展現出物理與光學特性是自然界材料所無法展現的性質。其中，Pendry等科學家提出用來展現負透磁率與高頻磁子的人造磁性原子-隙環共振器一直是最常設計來產生磁特性的超材料。一般而言，其基態的共振模態行為是透過等校的電容-電感電路概念來解釋，並且其複數個共振響應的特性可以藉由駐玻式電漿共振響應模型來闡述並預估其所對應的響應波長。更重要的是，此共振模態對隙環共振器周圍介質環境很敏感，特別是當生物分子結合於隙環共振器的表面時，其共振模態的頻率會移動，使其可作為即時的、不需標定的折射率細胞生物感測器。
    在過去，我們實驗室的研究中發現利用隙環共振設計的人造奈米結構超材料來觀察人體幹細胞中的折射率分佈。由於超材料的輻射光譜會隨著所處環境而改變，因此可直接由掃描每個位置的超材料得到人類幹細胞中的胞器影像。因為超材料不具毒性，觀測時也毋須侵入細胞，因此可作為即時影像且毋需染色標定的細胞生物顯微鏡。在本文的研究中，我們將超材料顯微鏡用於觀察活體細胞影像。藉由裝設FPA (focal plane array)之傅立葉轉換紅外線光譜儀量測結果得知，超材料顯微技術具備折射率感測器的功能，同時也可以得到近紅外折射率生物影像。
    我們初步實現了活體細胞影像量測的可能性。比起目前的生物切片影像，超材料顯微技術可以降低活體細胞損傷，直接觀測折射率影像；超材料顯微技術可以做到快速，即時，低破壞的生物顯微平台。由於目前活體細胞影像的擷取技術仍有相當大的挑戰空間，有效地改善影像擷取技術有助於提升活體細胞樣本的存活率。對於未來幹細胞分化研究，細胞內部物質鑑定，細胞內部藥物傳遞及細胞膜間分子交換觀測，有相當助益。

    Abstract 2 摘要 4 Acknowledgment 5 Content 6 List of Figures 8 List of Tables 11 Chapter 1 Introduction 12 1.1 Live Cell Imaging 12 1.2 Motivation 13 Chapter 2 Literature Review 15 2.1 The Background of Label technique 16 2.2 Microscopy 17 2.2.1 Light Microscopy 17 2.2.2 Conventional Fluorescence Microscopy 18 2.2.3 Confocal Microscopy 18 2.3 Synchrotron-Based Infrared Microscopy 19 2.3.1 Focal Plane Array 21 2.4 ATR-Based Microscopy 22 2.5 Split Ring Resonators Microscopy 23 2.5.1 Multi-Functional Split Ring Resonators 23 2.5.2 Intracellular Bio-imaging 32 2.6 Cell Line 38 2.6.1 HeLa Cell 38 Chapter 3 Experimental Procedure 40 3.1 CST Microwave Studio Simulations Methods 40 3.2 Sample Fabrication 40 3.2.1 Electron Beam Lithography (EBL) Process 40 3.2.2 E-gun Evaporation of Gold & Lift-off 42 3.2.4 Surface Modified 43 3.3 Cell Growth on Silicon Substrate with SRR Pattern 44 3.3.1 Cell Growth 44 3.4 Measurement 45 3.4.1 Micro-Fourier Transform Infrared Spectroscopy (μ-FTIR) & 45 Focal Plane Array (FPA) 45 3.5 Fluorescent method 46 3.5.1 4′-6-diamidino-2-phenylindole (DAPI) staining 46 3.6 MTT Assay 47 3.6.1 Cell viability test for hydrophilic layer 47 Chapter 4 Results and Discussion 49 4.1 Metamaterial 49 4.1.1 Introduction of matamaterial 49 4.1.2 Design 49 4.1.3 Surface Dressing Effect 54 4.1.4 Sensitivity 58 4.1.5 Detection length 59 4.2 HeLa cells 62 4.2.1 Introduction of HeLa cells 62 4.2.2 Cell Growth on Flask 63 4.2.3 Culturing Method for HeLa Cells on Silicon Substrate 64 4.3 Hydrophilic Layer 65 4.3.1 Methyl cellulose 65 4.3.2 Viability Test for Hydrophilic Layer (MTT assay) 65 4.4 Live Cells Imaging 67 4.4.1 The Definition of Spatial Resolution for Refractive Index Image 67 4.4.2 Intracellular, Live Cells Imaging 69 Chapter 5 Conclusion 73 Reference 74

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