指導教授：張煥宗臺灣大學：化學研究所陳亞拿Chen, Ya-NaYa-NaChen2014-11-252018-07-102014-11-252018-07-102014http://ntur.lib.ntu.edu.tw//handle/246246/261325本論文之主軸為以牛血清蛋白為模板合成新穎鈰／金複合金屬奈米團簇（BSA-Ce/Au NCs）。只需一步驟加熱合成，即可得到在325奈米的紫外光激發下，可以得到具有兩個放光波峰的螢光材料，峰值分別位於410奈米以及650奈米。410奈米的放光是源自於鈰（ＩＶ）離子與牛血清蛋白形成之錯合物，而650奈米為以蛋白質為模板合成的金奈米團簇之特徵放光波峰。實驗發現，410奈米的螢光強度會受到環境酸鹼值的改變而改變，且在鹼性環境下擁有較高的螢光強度，而650奈米放光峰的螢光強度並不受環境酸鹼值的影響，因此利用這兩個放光峰的光強度比值，可以量測出環境的酸鹼值。根據細胞毒性測試的實驗證明可以得知此複合式奈米材料具有相當好的生物相容性。此外，此材料之螢光十分穩定，其螢光強度在長時間紫外光的照射下及高鹽類濃度的環境下亦無顯著的改變。由此可知，牛血清蛋白-鈰／金複合金屬奈米團簇極適合做為生物系統中酸鹼值的螢光比值顯影劑。而藉由觀察本材料於海拉細胞內的共聚焦螢光顯微鏡影像，並調整海拉細胞環境的酸鹼值，發現此奈米團簇之放光性質仍然會因環境酸鹼值改變而有所不同，證明此奈米團簇感測器可以成功應用於觀察生物系統下酸鹼值的檢測。Novel bovine serum albumin (BSA) templated cerium/gold nanoclusters have been prepared using a facile one-pot heating synthetic method. The as-prepared nanoclusters, when irradiated with 325 nm UV light, possess two fluorescence emission peaks at 410 nm and 650 nm. The emission maximum at 410 nm is attributed to the contribution of Ce4+ complexed with BSA, and the emission at 650 nm is in the characteristic emission range of BSA-Au NCs. The peak at 410 nm is found to be sensitive to the pH values of the environment, while the emission at 650 nm is pH-inert. Therefore, BSA-Ce/Au NCs with emission peaks located at 410 nm and 650 nm can serve as a ratiometric pH sensor, using the ratio I410 / I650. Using protein as the synthetic template, BSA-Ce/Au NCs are found to be highly biocompatible, and its fluorescence intensities are stable against salt and constant irradiation. Confocal images of HeLa cells have proven that this probe can be used for detection in biological systems, and that altering the environmental pH of the cells, BSA-Ce/Au NCs could reflect the resulting intracellular changes.口試委員會審定書 # 誌謝 i 中文摘要 iv ABSTRACT v CONTENTS vi LIST OF FIGURES viii LIST OF TABLES xi Chapter 1 Introduction 1 1.1 Noble Metal Nanomaterials 2 1.2 Noble Metal Nanoclusters 3 1.3 Synthesis of Gold Nanoclusters 4 1.3.1 Dendrimer / Polymer-Stabilized Au NCs 4 1.3.2 Thiol-Capped Au NCs 5 1.3.3 Protein-Templated Au NCs 6 1.4 Applications of Gold Nanoclusters 6 1.4.1 Fluorescent Au NCs as Sensors 7 1.4.2 Fluorescent Au NCs as Bioimaging Agents 8 1.5 Research Motives 8 1.6 References 10 Chapter 2 One-Pot Synthesis of BSA-Ce/Au NCs as Ratiometric pH Sensors 27 2.1 Introduction 28 2.2 Experimental Section 29 2.2.1 Chemicals and Materials 29 2.2.2 Synthesis of BSA-Ce/Au NCs 30 2.2.3 Spectroscopic Characterization of BSA-Ce/Au NCs 30 2.2.4 Thermo Gravimetric Analysis (TGA) 31 2.2.5 MALDI-MS measurements 31 2.2.6 Cell Culture 32 2.2.7 Cell Viability Assay 32 2.2.8 Cellular uptake of BSA-Ce/Au NCs 32 2.2.9 Confocal fluorescence cell imaging 33 2.3 Results and Discussion 33 2.3.1 Optical properties of BSA-Ce/Au NCs 33 2.3.2 Formation of BSA-Ce complexes 35 2.3.3 One-Pot vs. Two-Step Synthesis of BSA-Ce/Au NCs 35 2.3.4 Precipitation and Re-Suspension of BSA-Ce/Au NCs 36 2.3.5 MALDI-MS Analysis of BSA-Ce/Au NCs 37 2.3.6 X-ray Photoelectron Spectroscopy Characterization of BSA-Ce/Au NCs 38 2.3.7 Thermogravimetric Analysis of BSA-Ce/Au NCs 38 2.3.8 pH-dependent properties and biocompatibility 39 2.3.9 HeLa cell fluorescence imaging 40 2.4 Conclusion 40 2.5 References 423161791 bytesapplication/pdf論文公開時間：2014/07/09論文使用權限：同意有償授權(權利金給回饋學校)鈰金奈米團簇細胞影像酸鹼感測器thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/261325/1/ntu-103-R01223101-1.pdf