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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳培菱 | |
dc.contributor.author | Shobhit Charan | en |
dc.contributor.author | 休伯西 | zh_TW |
dc.date.accessioned | 2021-05-17T09:22:51Z | - |
dc.date.available | 2015-02-16 | |
dc.date.available | 2021-05-17T09:22:51Z | - |
dc.date.copyright | 2012-02-16 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-01-16 | |
dc.identifier.citation | References
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6977 | - |
dc.description.abstract | 在過去幾十年的細胞生物學研究,螢光顯微分析技術已成為一項強而有力的細胞結構的影像呈現方法,只要對特定蛋白質單分子進行染色,就能用光學系統觀察此分子在細胞所在位置。然而,傳統螢光訊號不能提供詳細的分子訊息而且發光基團的光衰退會影響偵測的長時間穩定性和限制許多需要長時間偵測的應用。因而很多研究的努力方向在合成新材料以克服這些缺點;近幾來,高感度奈米金屬粒子光學影像技術已被廣泛使用在很多細胞或生醫應用。最近,先進的分子診斷材料用在體內影像和藥物釋的有電漿共振奈米金屬金或銀奈米粒子、生物可裂解奈米粒子和量子點。在這些不同光學影像技術中,金屬奈米粒子表面拉曼散射光譜具有多方面用途,當某一分子足夠接近金或銀組成的金屬奈米粒子,振動光譜強度能增加數個級數。以這些銀或金奈米粒子為組成的表面拉曼散射探測粒子可克服螢光標定的限制且已成功地應用在生醫應用。
有鑑於此,我們合成具表面拉曼散射活性的奈米探測粒子生物標的和呈像。奈米粒子的表面修飾可用生物性連接的方式逹成,其中涉及靜電吸引力,配位官能基交換或醯胺鍵結技術。 我們採用系統化改質奈米粒子表面,此拉曼散射活性奈米探測粒子可用於自細胞的層次到動物等級的檢測。首先,製備具磁性的銀奈米粒子做為體內光學影像,其探測粒子為銀包覆氧化鐵奈米粒子是結合核心氧化鐵的磁性和殼層奈米銀的表面拉曼散射。最外圍的官能基,我們進一步用具不同鹵素側基的苯硫官能基修飾以呈現清楚且不同的表面拉曼散射訊號。當這些被官能基化表面拉曼散射活性的探測粒子與3T3肌纖維母細胞一起培養,可發現細胞有效吸入這個探測粒子而且經由共焦影像系統,強化表面拉曼散射訊號的探測粒子可輕易被觀察。 第二、我們進一步發展以奈米銀為組成的表面拉曼散射探測粒子具脂質標的的表面修飾用在C.elegans體內影像偵測。在這次的工作,膠體銀奈米粒子表面非共價性連結脂質特定標的的Nile red 染料,所以探測粒子在有機體影像的分佈可被共焦拉曼散射顯微鏡觀察到。 此外,此拉曼散射探測粒子可獲得具現性影像,而在且在實驗中觀察到此探測粒子無毒性。 在更高等級的動物體內標的的研究,我們系統化發展多官能基表面拉曼散射活性金奈米柱狀體的探測粒子成功展現在癌細胞標的和CAL 27 xenografts體內影像。 基於研究成果,在偵測體內藥物釋放和光熱為基礎的應用,官能基化表面拉曼散射活性探測粒子是有前途的工具。 因此,我們發展表面拉曼散射活性的奈米粒子為組成的探測粒子用在體外和體內標的和呈像應用,在生醫的應用具有潛力。 | zh_TW |
dc.description.abstract | During the past few decades, fluorescence microscopy has been a powerful technique in the field of cell biology for imaging cellular structures with single molecular sensitivity. However, the fluorescence signals do not provide detailed molecular information, and the photo-bleaching effect often limits their applications. A lot of research efforts have been carried out to synthesize new materials to overcome these limitations. Recently, sensitive optical imaging technology using metal nanoparticles has been extensively used for many cellular and biomedical applications. Recent advancements in molecular diagnosis, in vivo imaging and drug delivery are obtained by using nanoparticles such as plasmon resonant nanoparticles (gold (Au) and silver (Ag)), biodegradable nanoparticles, quantum dots and surface-enhanced Raman scattering (SERS) tags. Among different optical imaging technology, SERS is a versatile technique which enhances the intensity of the vibrational spectra of a molecule by several order of magnitude when the molecule is in close proximity with metallic nanoparticles made of gold or silver. These silver and gold nanoparticles-based SERS probe have overcome the limitation of fluorescence labeling and have been successfully applied for biomedical applications.
In view of this, we have successfully synthesized SERS-active nanoparticles-based probe for targeting and imaging applications using biological systems. Surface engineering of nanoparticle were performed by using bioconjugation protocols involving electrostatic attraction, ligand exchange and amide-bond linkage techniques. Systematic improvement in the development of SERS-active nanoparticles-based probe was employed from the cellular level to animal model. At first, Silver nanoparticles-based probe was developed with added magnetic functionality for cellular sorting and imaging application. The developed probe comprises of silver-coated iron oxide nanoparticles to combine the magnetic property of iron oxide for sorting application and enhanced Raman scattering from metallic nanoparticles for in vitro optical imaging. Further functionalization of silver-coated iron oxide with benzenethiols of different halogen groups displayed clear and distinct Raman signature. When these functionalized SERS-active probe was incubated with 3T3 fibroblast cells, it was found that the cells could efficiently uptake the probe and due to the enhanced Raman signals, the probe can easily be observed through confocal imaging system. Secondly, we further developed SERS-active Silver nanoparticles-based probe for lipid targeting and in vivo imaging in C.elegans. In this work, colloidal silver nanoparticles were conjugated non-covalently with lipid specific Nile red dye and the images of organism were subsequently monitored through confocal Raman and conventional confocal microscope. Additionally, the images obtained by Raman probes were very reproducible and no toxicity was observed during the experiment. For in vivo targeting in higher animal models such as xenografts, the systematic development of SERS-active Gold nanorods-based probe for tumor targeting and in vivo imaging using CAL 27 xenografts was successfully performed. Based on our results, functionalized SERS-active probe can be promising tools for in vivo detection, drug delivery and photothermal-based applications. Hence, we are able to develop SERS-active nanoparticle-based probes for in vitro and in vivo targeting and imaging applications, which can be a potential candidate for biomedical applications. | en |
dc.description.provenance | Made available in DSpace on 2021-05-17T09:22:51Z (GMT). No. of bitstreams: 1 ntu-101-D95223035-1.pdf: 6236072 bytes, checksum: aada28e214e228f6d552f6a86188cb4e (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | Table of Contents
Chapter 1. Introduction 1.1 Nanomaterials…………………………………………………………………. 1 1.2 Noble metal nanoparticles…………………………………………………… 3 1.3 Surface modification of nanoparticles for biological application…… 4 1.4 Optical imaging using SERS technique……………………………………… 9 1.5 Basic concept of SERS spectroscopy………………………………………….. 10 1.6 Aim and scope of dissertation…………………………………………………. 14 Chapter 2. Magnetic SERS-active Silver Nanoparticles-Based Probe for Cellular Labeling and Imaging in NIH3T3 cells 2.1 Introduction……………………………………………..................................... 17 2.2 Experimental Methods…………………………………………………………. 18 2.2.1 Synthesis of Fe3O4@Ag core shell nanoparticles………………………... 18 2.2.2 Procedure for fabricating Fe3O4@Ag X(C6H4)SH; X=Br Cl, I…………. 19 2.3 Characterization………………………………………………………………... 19 2.4 Results and discussion…………………………………………………………. 19 2.5 Conclusion……………………………………………………………………... 26 Chapter 3. SERS-active Silver Nanoparticles-Based Probe for Lipid Targeting and In vivo Imaging in Caenorhabditis elegans (C.elegans) 3.1 Introduction……………………………………………………………………. 27 3.2 Experimental Methods…………………………………………………………. 28 3.2.1 Synthesis of Lipid targeting Probe (Nile red-coated Ag NPs)……………. 28 3.2.2 Preparation of C.elegans specimen (animal model)………………………. 29 3.2.3 Toxicity assessment……………………………………………………….. 29 2.2.4 In vivo imaging method……………………............................................... 30 2.3 Characterization……………………………………………………………….. 30 2.4 Results and discussion………………………………………………………… 31 2.5 Conclusion……………………………………………………………………... 41 Chapter 4. Multifunctional SERS-active Gold Nanorods-Based Probe for In vivo Localized Targeting and NIR-thermal Imaging in CAL 27 Xenografts 4.1 Introduction………..…………………………………….................................... 42 4.2 Experimental Methods………………………………………………………….. 45 4.2.1 Materials……………………………………………………………… 45 4.2.2 Preparation of Au seeds………………………………………………. 46 4.2.3 Preparation of growth solution and nanorods………………………… 46 4.2.4 PSS-coated gold nanorods…………………………………………… 46 4.2.5 PAA-coated gold nanorods…………………………………………… 47 4.2.6 PEG-coated gold nanorods…………………………………………… 47 4.2.7 Chitosan-coated gold nanorods……………………………………….. 47 4.2.8 Anti-EGFR-conjugated gold nanorods……………………………….. 48 4.2.9 Stability of anti-EGFR-conjugated gold nanorods…………………… 48 4.2.10 Quantification of antibody /nanoparticles by BCA assay……………. 49 4.2.11 Cytotoxicity Assay………………………………………………….. 50 4.2.12 Cellular uptake by ICP……………………………………………… 50 4.2.13 In vivo toxicity evaluation………………………………………….. 51 4.2.14 Biodistribution studies and In vivo NIR thermal imaging…………... 52 4.2.15 Fabrication of SERS-active CO-GNR probe 52 4.2.16 Statistical analysis…………………………………………………… 53 3.3 Characterization…………………………………………………………….. 53 3.4 Results and discussion……………………………………………………… 53 3.5 Conclusion…………………………………………………………………. 72 Chapter 5. Future outlook ………………………………………... 73 List of Publications………………………………………………... 74 References…………………………………………………………. 75 | |
dc.language.iso | en | |
dc.title | 發展奈米材料表面修飾技術於定域化標的細胞吸收的
生物影像應用 | zh_TW |
dc.title | Development and Surface Engineering of Nanomaterials for Cellular Uptake, Localized Targeting and Bioimaging Applications | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-1 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 李弘文 | |
dc.contributor.oralexamcommittee | 吳漢忠,李德章,董奕鍾 | |
dc.subject.keyword | 奈米粒子,表面工程,表面拉曼散射活性奈米探測粒子,標的,光學影像, | zh_TW |
dc.subject.keyword | nanoparticles,surface engineering,SERS-active nanoparticles-based probe,targeting,in vivo imaging, | en |
dc.relation.page | 90 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2012-01-16 | |
dc.contributor.author-college | 理學院 | zh_TW |
dc.contributor.author-dept | 化學研究所 | zh_TW |
顯示於系所單位: | 化學系 |
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