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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18632完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張煥宗(Huan-Tsung Chang) | |
| dc.contributor.author | Li-Wei Chuang | en |
| dc.contributor.author | 莊禮維 | zh_TW |
| dc.date.accessioned | 2021-06-08T01:16:04Z | - |
| dc.date.copyright | 2020-09-16 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-08-11 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18632 | - |
| dc.description.abstract | 氧化還原及酸鹼反應為影響螢光探針光學性質重要的機制。螢光碳奈米材料由於具有可調控之表面官能基，使其對局部刺激能快速的反應，可作為良好的檢測及顯影工具。於本篇研究中，我們發展一種新型碳奈米粒子 (量子產率 = 10%)，可用於偵測酸鹼值以及次氯酸根離子。此材料設計以間苯二胺為主體提供pH應答性，並結合維生素C多含氧官能基團，改善水溶性及增添氧化還原應答性的功能。因此，此碳奈米粒子可透過螢光變化作為pH探針及 ”turn-off” 探針於中性環境偵測次氯酸。研究結果發現其螢光於酸鹼值5.5至8.5具有線性關係 (R2 = 0.989)，而於中性環境下，對次氯酸根濃度的檢測範圍為0.125–1.25 μM，偵測極限為0.029 μM。此外，我們更進一步將此碳奈米粒子應用於細胞顯影，因具備正電之表面及奈米尺度，賦予其良好的細胞輸送效率，使其成功應用於細胞顯影，並偵測細胞pH值變化及次氯酸。基於以上優異特性，此螢光碳奈米粒子在未來有潛力作為用於細胞偵測酸鹼值及次氯酸根之螢光奈米碳針。 | zh_TW |
| dc.description.abstract | Acid-base and redox reactions are important mechanisms that affect the optical properties of fluorescent probes. Fluorescent carbon nanoparticles which possess tailored surface functionality enable a prompt response to regional stimuli, offering a useful platform for detection, sensing and imaging. In this study, carbon nanoparticle (CNP) was developed as a novel nanoprobe (quantum yield = 10%) for detection of pH and hypochlorite. In the design of CNP, m-phenylenediamine was chosen as the major component of CNP for pH responsiveness, while ascorbic acid which possesses many oxygen-containing groups was incorporated to generate favorable functionalities for improved water solubility and additional response toward redox reactions. Thus, the CNP could serve as a pH probe and a turn-off sensor toward hypochlorite at neutral pH through fluorescence change. The as-prepared CNP exhibited a linear fluorescence response over the pH ranges from pH 5.5 to 8.5 (R2 = 0.989), and over the concentration range of 0.125–1.25 μM for hypochlorite. Meanwhile, the detection limit (LOD) of hypochlorite was calculated to be 0.029 μM at neutral pH. In addition, the CPN was further applied to the cell imaging. The positively charged surface and nanoscale dimension of the CNP caused the efficient cellular delivery of the CNP. The CNP was successfully used to cell imaging and sensitive detection of hypochlorite as well as pH changes in biological system. Given these desirable performances, the as-synthesized fluorescent CNP shows great potential as an optical nanoprobe for cellular sensing of pH values and hypochlorite concentration in the future. | en |
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| dc.description.tableofcontents | Contents 國立台灣大學碩士學位論文口試委員會審定書 I 誌謝 II 中文摘要 IV Abstract V Contents VI Figure content X Table content XIII Chapter 1. Introduction 1 1.1 Carbon dots 1 1.1.1 Preface 1 1.1.2 Formation methods of carbon dots 1 1.1.2.1 “Top-down” methods 2 1.1.2.2 “Bottom-up” methods 2 1.1.3 Carbonized polymer dots 2 1.1.4 PL mechanisms of carbon dots 3 1.1.4.1 The quantum confinement effect 3 1.1.4.2 Surface state 4 1.1.4.3 Molecule state 4 1.1.4.4 Crosslink-enhanced emission (CEE) effect 5 1.2 pH-responsive fluorescent carbon dots 5 1.3 Redox responsive fluorescent carbon dots 6 1.4 Research motives 7 Chapter 2. Experimental section 8 2.1 Chemicals 8 2.2 Cell culture 9 2.3 Preparation of ROS/RNS 9 2.3.1 Preparation of O2- 9 2.3.2 Preparation of •OH 9 2.3.3 Preparation of ONOO- 10 2.3.4 Preparation of NaClO, H2O2 10 2.4 Preparation of mPA CNPs, mPD C-dots, and AA NS 10 2.4.1 Preparation of mPA CNPs 10 2.4.2 Preparation of mPD C-dots 10 2.4.3 Preparation of AA NS 11 2.5 Characterization of carbon materials 11 2.5.1 Quantum yield 11 2.5.2 Characterization of carbon dots 12 2.6 pH response of mPA CNPs 13 2.7 Fluorescence images of mPA CNPs 14 2.8 PL response of mPA CNPs, mPD C-dots, AA NS toward hypochlorite 14 2.9 PL response of mPA CNPs and DCFH-DA toward anti-oxidants and reactive oxygen species 14 2.10 Detection of hypochlorite with mPA CNPs 14 2.11 Cell Viability 15 2.12 Cell images, and flow cytometry of mPA CNPs under various pH conditions 15 2.13 Real-time observation of cell images and quantification with flow cytometry under various concentrations of hypochlorite 16 Chapter 3. Results and discussion 17 3.1 Characterization of mPA CNPs 17 3.2 Optical properties of mPA CNPs 20 3.3 pH-responsiveness of mPA CNPs 22 3.4 Mechanisms of fluorescence change of mPA CNPs under various pH conditions 23 3.5 pH-responsiveness and reversibility of mPA CNPs 25 3.6 Hypochlorite-responsiveness and PL quenching mechanisms of mPA CNPs 25 3.7 Detection of hypochlorite with mPA CNPs 27 3.8 Bio-imaging of mPA CNPs under various pH values 27 3.9 Bio-imaging of mPA CNPs under various concentrations of hypochlorite 28 3.10 Conclusion 29 Figures 31 Tables 51 References 57 ‘ Figure content Scheme 1. Scheme representations of (a) pH reversibility of mPA CNPs (b) PL quenching of mPA CNPs by ClO- 31 Fig. 1 TEM images of (a) mPA CNPs (inset: size distribution and HRTEM image) (b) mPD C-dots (inset: size distribution) (c) AA NS 32 Fig. 2 (A) HR-TEM image and (B) SEM image of mPA CNPs 33 Fig. 3 (A) XPS survey (B) C1s (C) O1s (D) N1s spectra of (a) mPA CNPs (b) mPD C-dots (c) AA NS 34 Fig. 4 FT-IR spectra of (A) AA (B) mPD (C) mPA CNPs (D) AA NS (E) mPD C-dots (ν: stretching, δ: bending) 35 Fig. 5 (A) UV-vis spectra and (B) fluorescence spectra of (a) mPD C-dots and (b) mPA CNPs (inset: photo images, from left to right 170, 200, 230, and 260℃) and (c) various ratio of mPA CNPs (inset: photo images, from left to right mPD: AA= 2:1, 1:1, 1:2) Note: mPD C-dots (λex: 320 nm), mPA CNPs (λex: 400 nm) (λex denotes excitation wavelength) 36 Fig. 6 (A) UV-Vis, strongest fluorescence emission spectra and (B) Fluorescence spectra (λex: from 320 nm to 480 nm) of (a) mPA CNPs (b) mPD C-dots (c) AA NS (insets of A-a, A-b, and A-c were photo images of three materials (1 mg/ mL)) (λex denotes excitation wavelength) 37 Fig. 7 (A) Fluorescence spectra (inset: photo images) of mPA CNPs (1 μg/ mL) (B) UV-vis spectra (inset: photo images) of mPA CNPs (50 μg/ mL) under pH 3, 5, 7, 9, and 11 at excitation of 400 nm (C) pH titration curves of (a) AA NS (λex: 340 nm, λem: 430 nm), (b) mPD C-dots (λex: 400 nm, λem: 500 nm) (c) mPA CNPs (λex: 400 nm, λem: 500 nm) (D) mPA CNPs: the linearity of PL intensity versus pH values from pH 5.5 to 8.5 (λex denotes excitation wavelength, and λem denotes emission wavelength) 38 Fig. 8 Fluorescence images of mPA CNPs (10 μg/mL) exposure time: 1/7 s (λex: 360-370 nm, λem: 420 nm) under (A) pH 3 (B) pH 5 (C) pH 7 (D) pH 9 (E) pH 11 (λex denotes excitation wavelength, and λem denotes emission wavelength) 39 Fig. 9 (A) pH-reversible fluorescence intensity, (B) pH-reversible apparent color and (C) fluorescence of mPA CNPs (50 μg/mL) with 0.1 M HCl and 0.1 M NaOH added alternately. 40 Fig. 10 Variations of fluorescence intensity (a) without (b) with NaClO exerted on (A) mPA CNPs (λex: 400 nm, λem: 500 nm), (B) mPD C-dots (λex: 340 nm, λem: 430 nm), and (C) AA NS (λex: 340 nm, λem: 430 nm). 41 Fig. 11 FT-IR spectra of various materials (A) without (B) with NaClO addition, (a) mPA CNPs (b) mPD C-dots (c) AA NS 42 Fig. 12 (a) PL response of mPA CNPs (5 μg/ mL) toward various reagents under pH 7.4 (NaClO and ONOO- was 10 μM, while H2O2, O2-,˙OH, NaNO2, (NH4)2C2O4, and NaBH4 were 100 μM). (b) Linearity of fluorescence change of mPA CNPs (0.05 μg/ mL) versus concentration of NaClO from 0.125 μM to 1.25 μM under pH 7 (phosphate buffer: 20 μM) F denotes PL intensity of mPA CNPs with ROS/ RNS, while F0 denotes PL intensity of mPA CNPs without ROS/RNS. 43 Fig. 13 Cell viability for Tramp C1 cells in the presence of a series of concentrations of mPA CNPs. 44 Fig. 14 Tramp C1 cell images under pH 5.4, 6.4, 7.4, and 8.4. From left to right were different fields: bright field, blue channel for positioning with hochest, green channel with mPA CNPs (λex: 360-490 nm, λem: 520 nm). The scale bar denoted 20 μm. 45 Fig. 15 (A) Tramp C1 cells distribution profiles under various pH values (a) Blank (without mPA CNPs staining), (b) pH 5.4, (c) pH 6.4, (d) pH 7.4, (e) pH 8.4 and (B) Fluorescence intensity profiles of collected cells (C) Relative fluorescence intensity from (B) (F0 denotes PL intensity under pH 5.4, and F denotes PL intensity under various pH values) 46 Fig. 16 Images of Tramp C1 cells treated with NaClO (a) 0 μM, (b) 10 μM, after mPA CNPs staining. The top image is for positioning (hochest), then from second images to the last are the bright field (left) and fluorescence images (right) (from 0 to 30 min after treated with NaClO) (λex: 360-490 nm, λem: 520 nm). The scale bar denotes 32 μm. 47 Fig. 18 PL intensity quantification of Tramp C1 cells treated with various concentrations of NaClO (0-10 μM). (A) Fluorescence intensity profiles and (B) the relative fluorescence intensity of collected Tramp C1 cells stained with (a) mPA CNPs (b) DCFH-DA 49 Fig. 19 PL response of mPA CNPs (5 μg/ mL) and DCFH-DA (25 μM) toward various ROS/RNS under pH 7.4 (NaClO and ONOO- was10 μM, while H2O2, O2-,and˙OH were 100 μM. F denotes PL intensity of mPA CNPs after reacting with ROS/RNS, while F0 denotes PL intensity of mPA CNPs without ROS/RNS) 50 Table content Table. 1 Quantum yield of as-prepared materials 51 Table. 2 Lifetime of mPA CNPs (1 μg/mL) under various pH conditions 52 Table. 3 Measurement of zeta potential and size with mPA CNPs (1 μg/mL) 53 Table. 4 Reduction potential of antioxidant and ROS 54 Table. 5 Comparison of developed hypochlorite sensors 55 Table. 6 Comparison of developed pH sensors 56 | |
| dc.language.iso | en | |
| dc.title | 合成用於偵測細胞內pH值與次氯酸濃度的螢光碳奈米粒子 | zh_TW |
| dc.title | Synthesis of fluorescent carbon nanoparticles for cellular sensing of pH values and hypochlorite concentration | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃志清(Chih-Ching Huang),黃郁棻(Yu-Fen Huang),胡焯淳(Cho-Chun Hu) | |
| dc.subject.keyword | 螢光碳奈米粒子,細胞顯影,pH探針,ROS選擇性,次氯酸根探針, | zh_TW |
| dc.subject.keyword | fluorescent carbon nanoparticle,cell imaging,pH probe,ROS selectivity,hypochlorite probe, | en |
| dc.relation.page | 65 | |
| dc.identifier.doi | 10.6342/NTU202002994 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2020-08-12 | |
| 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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