本論文的重點在於設計可調控特性的多功能奈米材料，並應用在生物醫學和生物感測器中。該研究探討奈米顆粒、奈米團簇和奈米片的合成策略，並同時越過了材料的挑戰，從而獲得了理想的光電特性。這些奈米材料用於檢測生物流體和癌症生物成像中的疾病生物標記，鹼性磷酸酶、肌酸激酶和過氧化氫。第三章以剝落法合成二維α-MoO3-x奈米薄片，在近紅外條件下獲得等表面電漿峰。與過氧化氫相互作用後，最初為藍色的α-MoO3-x溶液被氧化，從而改變其氧化態以形成α-MoO3。此奈米薄片的顏色變化從藍色轉變為灰藍色，吸收光譜也有明顯變化。利用其光學性質，應用在檢測生物流體中的過氧化氫（H2O2）。第4章討論了對鉬的量子限制效應，從而獲得了一種螢光鉬奈米團簇（MoNCs）。該反應系統的化學成分在優化的條件下（具有水和pH穩定性）使球形顆粒中快速出現綠色螢光。由於這些特性，MoNC被用於HaCaT和A549癌細胞的成像。第5章詳細介紹了一種新穎的方法，可合成獨特的無模板穩定蛋白的金奈米立方體（PSGNC），用於感測和定量生物流體中的癌症生物標記物鹼性磷酸酶（ALP）。 PSGNC的波長最大發射值與ALP催化反應終產物即405 nm的對硝基苯酚（p-NP）的最大吸收值重疊。該光譜重疊在基於螢光內濾效應（IFE）下，在生物流體中檢測ALP。第6章探討在ATP存在下，聚集半胱胺（Cys）功能化金奈米粒子(GNPs)的獨特方法，用於有效檢測血清中的心臟生物標記物肌酸激酶（CK-MM）。帶正電的Cys-GNP（磚紅色）在存在帶負電的ATP（藍色）下聚集，但是當將CK-MM添加到溶液中時，可防止轉化。這種相互作用為鑑定血清中CK-MM奠定了基礎。第7章說明了鑭系元素的螢光性質在生物感測中的適用性。該研究詳細介紹基於螢光的H2O2檢測系統。利用乾磨和濕磨相結合，然後進行退火和煅燒，得到結晶且高度有序的鈰-鉬-銪（CME）鈣鈦礦。所獲得的粒徑出乎意料的小（<100 nm），在近紅外（NIR）範圍內具有水分散性和強烈的紅色螢光。
在溶液中加入極少的H2O2的過程中，改變了金屬氧化物鍵，導致螢光信號增強，進一步利用此螢光反應檢測生物流體中的H2O2。

The main focus of this thesis is to design multiple functional nanomaterials with tunable properties for their applications in biomedicine and biosensor. The research projects explore the synthesis strategies for nanoparticles, nanoclusters, and nanosheets while pushing beyond the material challenges to obtain desirable optoelectronic characteristics. These nanomaterials are further employed to detect disease biomarkers alkaline phosphatase, creatine kinase, and hydrogen peroxide in biological fluids and cancer bioimaging. Chapter 3 discusses the synthesis of two-dimensional α-MoO3-x nanoflakes by an exfoliation-based method to obtain plasmonic peaks in the near-infrared regime. α-MoO3-x initially blue-colored solution is oxidized after interaction with hydrogen peroxide, thereby changing its oxidation state to form α-MoO3. The change in the nanoflakes' oxidation state transforms from blue to a visually distinct hazy blue color with an apparent shift in the absorption spectrum. The optical property is explored in the detection of hydrogen peroxide (H2O2) in the biological fluid. Chapter 4 discusses about the quantum confinement effect on molybdenum to obtain one-of-a-kind fluorescent molybdenum nanoclusters (MoNCs). The reaction system's chemical composition resulted in the rapid emergence of green fluorescence from the spherical particle under optimized conditions with aqueous and pH stability. Owing to these characteristics, MoNCs were used for imaging of HaCaT and A549 cancer cells. Chapter 5 details a novel approach for synthesizing unique template free protein stabilized gold nanocubes (PSGNCs) for sensing and quantifying cancer biomarker alkaline phosphatase (ALP) in biological fluids. The wavelength emission maxima of PSGNCs overlap with the absorption maxima of the final product of ALP catalyzed reaction, i.e., p-Nitrophenol (p-NP) at 405 nm. This spectral overlap was used in an inner filter effect (IFE) based detection system for ALP in the biological fluid. Chapter 6 explores a unique method based on the aggregation of cysteamine (Cys) functionalized GNPs in the presence of ATP for effective detection of cardiac biomarker creatine kinase (CK-MM) in serum. Positively charged Cys-GNPs (brick red color) aggregate in the presence of negatively charged ATP (blue color), but the transformation is prevented when CK-MM is added to the solution. This interaction lays the foundation for identifying CK-MM in serum. Chapter 7 shows the applicability of intrinsic fluorescent property of lanthanides in bio-sensing. The research details the design of a fluorescence-based detection system for H2O2. A combination of dry and wet grinding followed by annealing and calcination resulted in crystalline and highly ordered cerium-molybdenum-europium (CME) perovskites. The particle size obtained was surprisingly small (<100 nm) with aqueous dispersity and intense red fluorescence in the near-infrared (NIR) regime. During its incubation with minimum H2O2 in the solution, the metal oxide bond was altered, resulting in an enhanced fluorescence signal. This fluorescent response was further extended to detect H2O2 in a biological fluid.

Table of Contents
Thesis/Dissertation validation letter …………………………………..………..…….…….. i
Acknowledgements ………………………...…………………………………….….…….. ii
Abstract (Chinese) ………………………………………………………………......….… vii
Abstract ……………………………………………………………..………...…......…..… ix
Table of Figures ……………………………………………………….………………….. xv
Table of Tables ……………………………………………………...…………...……..… xxi
Chapter 1
Introduction ……………………………………………………...………………………..... 1
1. Definition of nanomaterials ………………………………………………………...…... 1
2. History of nanomaterials ……………………………………………………………….. 1
3. Types of nanomaterials ……………………………………………………………….... 3
4. Properties of nanomaterials ……………………………………………………...…...… 4
5. Principles of nanomaterial synthesis ……………………...……………………...…..… 6
6. Principles of Biosensing …………………………………………...…………………… 9
7. Highlights of the upcoming chapter ………………………………………………...… 11
8. Reference ………………………………………………………………………...…… 13
Chapter 2
Instrumentation ………………………………………………………………………….... 20
1. UV-Vis spectroscopy ……………………………………………..………………....... 20
2. Fluorescence spectroscopy …………………………………………………………..... 22
3. Method for identifying limits of detection …………………………………………..… 24
4. Raman spectroscopy ………………………………………………………………..… 24
5. Electron micrographs ………………………………………………………………..... 26
6. X-ray photoelectron spectroscopy …………………………………………………..… 27
7. X-ray diffraction ……………………………………………………………………… 29
8. Cyclic voltammetry ………………………………………………………………....… 31
9. Reference …………………………………………………………………………...… 33
Chapter 3
Two dimensional α-MoO3-x nanoflakes as bare eye probe for hydrogen peroxide in biological fluids ………………………………………………………….……………………...…… 35
1. Introduction ………………………………………………………………………...…. 35
2. Materials and Methods …………………………………………………………...….... 37
3. Results and Discussions …………………………………………………………….… 40
4. Conclusion ....………………………………………………………………………..... 50
5. Reference ……………………………………………………………………………... 50
Chapter 4
Synthesis of fluorescent molybdenum nanoclusters at ambient temperature and their application in biological imaging …………………………………………………………. 60
1. Introduction ………………………………………………………………………….... 60
2. Materials and Methods …………………………………………………………...….... 62
3. Results and Discussions …………………………………………………………….… 66
4. Conclusion ....…………………………………………………………………………. 91
5. Reference …………………………………………………………………………....... 92
Chapter 5
Protein stabilized fluorescent gold nanocubes as selective probe for alkaline phosphatase via inner filter effect ……………………………………………………...…………...…...… 104
1. Introduction ……………………...…………………………………………...…….... 104
2. Materials and Methods ……………………………………….………………....….... 106
3. Results and Discussions ………………………………………..………..…….......… 108
4. Conclusion ....………………………………………………………………...…….... 117
5. Reference …………………………………………………………..……………....... 118
Chapter 6
Aggregation of cysteamine-capped gold nanoparticles in presence of ATP as an analytical tool for rapid detection of creatine kinase (CK-MM) ……………………….…………… 126
1. Introduction ………………………………………………………………………….. 126
2. Materials and Methods …………………………………………………………......... 128
3. Results and Discussions …………………………………………………………....... 131
4. Conclusion ...……………………………………………………………………….... 147
5. Reference …………………………………………………………………………..... 147
Chapter 7
Ce1.0Mo0.15Eu0.05Ox aqueous perovskites for stable NIR-emission and its sensitivity towards hydrogen peroxide …………………...………………………………………………..… 155
1. Introduction ………………………………………………………………………….. 155
2. Materials and Methods …………………………………………………………......... 157
3. Results and Discussions ……………………………………………………………... 160
4. Conclusion ...……………………………………………………………………….... 180
5. Reference ……………………………………………………………………………. 181
Conclusion ……………………………………………………….…………………….... 190
Appendix ……………………..…..…………………………………………………...… 192
 
Table of Figures
Chapter 1
Fig. 1.1: A photograph of the Lycurgus cup &amp; Michael Faraday’s gold colloids ……….… 3
Fig. 1.2: Different dimensions of nanomaterials ……………………………...………….... 4
Fig. 1.3: Size dependent change in plasmonic gold nanoparticles (Left); Fluorescent CdSe quantum dots, (Right)............................................................................................................. 6
Fig. 1.4: (A) Quantum confinement effect; (B) Schematic depicting the localized surface plasmon resonance of surface delocalized electrons …………………………………......… 7
Fig. 1.5: Schematics of bottom-up and top-down approach for synthesis of nanomaterials through various chemical treatments ………………………………………………..…........ 8
Fig. 1.6: Schematic representation of (A) Ostwald ripening; (B) Digestive ripening …....... 9
Fig. 1.7: Schematics representing lateral flow immunoassay using gold nanoparticles…... 11
Fig. 1.8: Schematics representing (A) Forster resonance energy transfer (FRET); (B) Inner Filter effect (IFE) …............................................................................................................. 12

Chapter 2
Fig. 2.1: Principle of absorption spectroscopy depicting the incident and transmitting radiation with the path length covered by the light through the solution and the concentration gradient ………………………………………………………………………………...…. 21
Fig. 2.2: The principle of photoluminescence depicting fluorescence and phosphorescence phenomenon under the influence of incident excitation radiation ……………………….... 23
Fig. 2.3: Schematic depicting the various scattering of light after interaction with nanomaterial solution …………………………..…………………………………………. 25
Fig. 2.4: Schematic of transmission and scanning electron microscope ………………..… 27
Fig. 2.5: Schematic representing the XPS...…………………………...………………...… 28
Fig. 2.6: Schematic representing the principle of Bragg’s law used in X-ray diffraction..... 30
Fig. 2.7: Schematics depicting cyclic voltammetry electrochemical cell and a demonstrative voltammogram showing the anodic and cathodic peak potential ………………..………... 32

Chapter 3
Scheme 3.1: Schematic representation of the detection system at a glance ………….…… 41
Fig. 3.1: Graphical representation: Comparing the sensitivity of colorimetric methods earlier reported for detection of H2O2 ……………………………………...…………………...… 42
Fig. 3.2: (A-B) TEM characterization of α-MoO3-x nanoflakes; (B-C) XPS analysis of the nanoflakes before and after addition of H2O2 …………………………………..…………. 43
Fig. 3.3: (A) UV-Vis spectra and (B) Day-light photograph of H2O2 concentration dependent color change of α-MoO3-x; (C-D) Concentration dependent change in absorption spectra .. 45
Fig. 3.4: UV-Vis absorbance spectra of α-MoO3-x¬ in presence of varying concentration of urine spiked H2O2 ………………………………………………………..……………...… 46
Fig. 3.5: (A) Relative intensity I/I0 vs. Concentration of H2O2 spiked in biological fluid. (B) Day-light photograph of color change in H2O2 spiked biological sample ………………… 47
Fig. 3.6: (A) Selective assay of OH• radicals generated using Fenton Reaction; (B) Comparing percent recovery of H2O2 in urine samples conventional method …..………... 48
Fig. 3.7: (A) Representative of “zone of inhibition” recorded in petri dish; (B) Relative absorbance showing selective analysis of urine samples…………………………..………. 49

Chapter 4
Scheme 4.1: Schematic representation of the synthesis of molybdenum nanocluster .....… 67
Fig. 4.1: (A) UV-Vis Spectroscopy; (B) Fluorescence spectroscopy; (C-D) Time-dependent observation using UV-Vis spectroscopy and fluorescence spectroscopy …………..……... 70
Fig. 4.2: (A) TRPL; and (B) pH stability of MoNCs ……………………………………... 71
Fig. 4.3: Synthesis of MoNC at different pH solutions of NaOH ……………...…….....… 72
Fig. 4.4: Salt stability of MoNC (A) absorbance and (B) fluorescence spectroscopy ……. 73
Fig. 4.5: Physical characterizations involving (A-C) TEM; (D) Dynamic Light Scattering measured for 2 ml solution; (E) FTIR spectroscopy …………………………………….… 74
Fig. 4.6: Zeta potential spectrum of MoNCs (-77.75 mV) ………….…………………….. 75
Fig. 4.7: XPS spectra of MoNCs …………………………………………….……………. 76
Fig. 4.8: (A) XRD pattern; (B) Cyclic voltammogram of MoNCs ………….……..…...… 78
Fig. 4.9: Cyclic voltammogram of MoCl3 ……………………………………………….... 78
Fig. 4.10: Photostability of MoNCs under continuous light exposure …………...…..…… 83
Fig. 4.11: DLS and Zeta potential of MoNCs after incubating with serum ……………..... 84
Fig. 4.12: Confocal images of the MoNCs entering the cell cytoplasm ………………...… 84
Fig. 4.13: Cell viability assessment of MoNCs against HaCaT cells. …………………….. 85
Fig. 4.14: MoNC internalization of A549 cells (A-C); (D) 3D tomographic image …….... 85
Fig. 4.15: MoNC internalization of RPTEC cells (A-C); (D) 3D tomographic image ...…. 87
Fig. 4.16: Cell viability analysis of A549 cells ……………..……….……………….….... 88
Fig. 4.17: Cell viability analysis of Kidney epithelial cells ……………………………..... 88
Fig. 4.18: Percent cellular uptake of MoNCs in A549 cells ……………………………..... 90
Fig. 4.19: Quantitative cellular uptake of MoNCs …………………………..…………..... 91

Chapter 5
Fig. 5.1: (A) UV-Vis absorbance (Inset: TEM representative); (B) Fluorescence spectroscopy of PSGNC …………………………………………………………………. 110
Fig. 5.2: Effect of different saline concentration on detection system ………………....… 111
Fig. 5.3: Effect of different pH on the sensitivity of detection system ………….......….... 111
Fig. 5.4: Effect of different metals on the detection system ……………………......….… 111
Scheme 5.1: Schematic representation of working principles for sensing alkaline phosphatase in the presence of p-Nitrophenyl phosphate (p-NPP) …………………….… 114
Fig. 5.5: (A) Concentration dependent assay of ALP; (B) Relative fluorescence intensity 115
Fig. 5.6: (A) Concentration dependent assay in serum; (B) Relative fluorescence intensity ……………………………………………………………………...………....... 116
Fig. 5.7: Selectivity of the sensing system …………………………………………….… 117

Chapter 6
Scheme 6.1: Schematics depicting reaction between gold nanoparticles and CKMM….... 133
Fig. 6.1: TEM of Cys-GNP (A) and aggregated nanoparticles (B); (C) Photograph of nanoparticles; (D) UV-Vis absorbance spectra; (E) DLS of bare Cys-GNP (Inset: Zeta potential); (F) Cys-GNPs in presence of CK-MM and ATP …………………..…............. 135
Fig. 6.2: Optical response of other components in the reaction system ………………….. 137
Fig. 6.3: Inhibition of ALP for CK-MM activity ……………………………………..…. 138
Fig. 6.4: (A) pH stability of the detection system; (B) Relative absorbance of A532/A700 …140
Fig. 6.5: (A) Salt stability of the reaction system; (B) Relative absorbance of A532/A700 .. 140
Fig. 6.6: (A) Varying ATP concentrations; (B) Relative absorbance plot of A532/A700 ..… 141
Fig. 6.7: (A) Detecting varying CKMM concentrations (Inset: A532/A700 Vs concentration of CK-MM; (B) Day-light photograph of Cys-GNPs in presence of CK-MM ..……………. 142
Fig. 6.8: (A) A532/A700 Vs Concentration of CK-MM (Inset: Optical response of different concentrations of CK-MM in presence of serum); (B) Day-light photograph of Cys-GNPs in presence of CK-MM. ……………………………………………………………………. 143
Fig. 6.9: Selectivity of the method …………….………………...…………..…………... 146

Chapter 7
Scheme 7.1: Stepwise synthesis scheme of CME perovskites; predicted crystal structure; Day light and UV-lamp exposed photographs of the as-synthesized CME perovskites …. 161
Fig. 7.1: (A) High resolution TEM image; (B) SAED pattern; (C-D) Magnified image of the HRTEM image (Inset: FFT representative of the selected area)...…………………..……. 162
Fig. 7.2: (A) TEM image; (B) Histogram showing the particle counts; (C-D) TEM image and Energy dispersive x-ray spectroscopy (EDS) ………………………………………….… 163
Fig. 7.3: (A) X-ray diffraction spectrum; (B) The predicted crystal structure ………........ 165
Fig. 7.4: (A) UV-Vis absorption and fluorescence spectrum (Inset: photographic representation of perovskites); (B) Raman shifts ……………………………….……...… 166
Fig. 7.5: (A) Solvent treatment in bright daylight and UV-lamp exposure photographs; (B-C) Fluorescence spectra ………………………………………………….....................… 167
Fig. 7.6: pH stability of CME perovskites (A) UV-Vis spectra; (B) 3D fluorescence spectra; (C) Relative fluorescence intensity …………………………………………………....… 168
Fig. 7.7: (A) Comparing fluorescence behavior in CE, ME and CME (at λex = 370 nm); (B) Raman spectra of CE, ME and CME ……………………………….………………...….. 170
Fig. 7.8: Time resolved photoluminescence of CME perovskites ……………..………… 171
Fig. 7.9: X-ray photoelectron spectroscopy (XPS) of the synthesized CME perovskites... 172
Fig. 7.10: X-ray photoelectron spectroscopy (XPS) of ME ……………………………… 173
Fig. 7.11: X-ray photoelectron spectroscopy (XPS) of CE ………………………………. 173
Fig. 7.12: 3D Raman spectra for CME treated with H2O2 …………….…….…………… 175
Fig. 7.13: XPS of CME perovskites after treatment with H2O2 ……………………….…. 175
Fig. 7.14: Cyclic voltammogram of (A) CME; (B) Hydrogen peroxide treated CME….... 176
Fig. 7.15: CME perovskites with various concentrations of H2O2. ……..…………...…… 177
Fig. 7.16: (A) Concentration dependent H2O2 detection; (B) Linear regression.……........ 178
Fig. 7.17: Graphical comparison of fluorescence-based detection methods of H2O2….... 178
Fig. 7.18: (A) Concentration dependent H2O2 detection in urine; (B) Linear regression ... 179
Fig. 7.19: Effect of free oxygen radicals on the CME perovskites …….………………… 180
 
Table of Tables
Chapter 3
Table 3.1: DLS studies of the α-MoO3-x nanoflakes in presence of various concentration of H2O2………………………………………………...………………………………..….… 44
Table 3.2: “Zone of inhibition” recorded in petri dish containing α-MoO3-x infused 0.8% agarose. Wells contain various concentration of H2O2…………………………………….. 49

Chapter 4
Table 4.1: Tabular representation of MoNCs particles size and zeta potential ……….….. 75
Table 4.2: Tabular comparison between synthesis of conventional MoS2 and the current MoNCs ………………………………………………………………………………….… 80

Chapter 5
Table 5.1: Comparison of the limit of detection of fluorescence-based detection of alkaline phosphatase …………………………………………...………………………………..... 113

Chapter 6
Table 6.1: Tabular illustration of control and sample groups shown in Fig. 6.1D ………. 136
Table 6.2: Tabular illustration of control and sample groups shown in Fig. 6.2 …..…..… 137
Table 6.3: Tabular illustration of control and sample groups shown in Fig. 6.3 ………… 139
Table 6.4: Tabular illustration of control and sample groups shown in Fig. 6.8 ………… 143
Table 6.5: Tabular comparison between strategies reported for detection of creatine kinase (CK) and its isoforms (MB and MM) …………………………………………………..... 144
Chapter 7
Table 7.1: Tabular details of the element’s composition of the CME perovskite nanoparticles …...…………………………………………………………………...…… 163

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Chapter 7
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9. Souza AE, Almeida Santos GT, Silva RA, et al. Morphological and structural changes of Ca xSr 1-xTiO 3 powders obtained by the microwave-assisted hydrothermal method. Int J Appl Cera