Madhuri Sharon, (Retd Director Reliance Industries LTD) PhD from Leicester University UK, Post-Doctoral Research from Bolton Institute of Technology U.K., is currently Director of NSN Research Centre for Nanotechnology & Bionanotechnology and Managing Director of Monad Nanotech as well as Adjunct-Professor University of Mumbai & Professor-Emeritus JJT University. She has published more than 130 papers, 4 books and 11 patents. Her research focuses on the synthesis, biosynthesis and application of various nanomaterials (graphene oxide, carbon dots, carbon nanomaterials and nanometals, SPIONS) in drug-delivery. Ashmi Mewada holds a PhD degree from Nagoya Institute of Technology, Japan. She is currently an Assistant Professor in the Department of Nanotechnology at Walchand College of Arts and Science, Solapur. She has more than 30 publications and works extensively in the field of carbon dots.

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• Preface1. Carbon Dots: Discovery, Synthesis and Characterization 11.1. Background 11.2. Introduction to QD 21.2.1. What is Quantum Mechanics? 41.2.2. Quantum Confinement 71.2.3. Discovery and History of Carbon Dots 81.3. Carbon QD and Graphene QD 91.4. Various Methods of Synthesis of Carbon Dots 101.4.1. Electrochemical Methods 111.4.2. Combustion and Thermal Oxidation Method 131.4.3. Hydrothermal Oxidation Method 151.4.4. Solvothermal Method 181.4.5. Laser Ablation of Graphite 181.4.6. Pulsed Laser Irradiation of Carbon Source 201.4.7. Arc Discharge Method 201.4.8. Plasma Treatment 211.4.9. Opening of Fullerene Cage 221.4.10. Ultrasonication Method 221.4.11. Microwave-Assisted Method 231.4.12. Chemical Methods 261.4.13. Supported Synthetic Procedure 261.4.14. Biogenic Method 281.5. Characterization of Carbon Dots 311.5.1. Microscopic Methods 321.5.1.1. SEM and TEM Characterization 321.5.1.2. AFM and STM Characterization 341.5.2. Spectroscopic Methods 351.5.2.1. UV-Vis Spectroscopy and its Application for Band Gap Determination 371.5.2.2. Fluorescence Spectrometry 371.5.2.3. Fourier Transform Infrared (FTIR) Spectroscopy 381.5.2.4. X-Ray Diffraction (XRD) Analysis 401.5.2.5. X-Ray Photoelectron Spectroscopy (XPS) 411.5.2.6. Dynamic Light Scattering/Photon Correlation Spectroscopy (DLS/PCS) 411.5.2.7. Dual Polarization Interferometry (DPI) 421.5.2.8. Raman Spectroscopy 431.5.2.9. Nuclear Magnetic Resonance (NMR) Spectroscopy 441.6. Summary 452. Properties of Carbon Dots 472.1. Introduction 472.2. Optical Properties 492.2.1. Absorbance 512.2.2. Photo-Induced Electron Transfer (PET) with CDs 522.2.3. Fluorescence/Photoluminescence (PL) 532.2.3.1. Multiphoton Excitation 602.2.3.2. Upconversion Photoluminescence 612.2.3.3. Lack of Blinking 632.2.3.4. Resistance to Photobleaching 642.2.4. Photocatalytic Property 652.3. Chemically Inert 672.4. Easy Functionalization 672.5. Water Solubility 682.6. Low Toxicity 682.7. Biocompatibility 692.8. Summary 703. Carbon Dots and Conjugates 713.1. Introduction 713.2. Why Conjugation of Carbon Dots? 743.3. Types of Carbon Dot Conjugates and Their Applications 763.3.1. Biogenic Compounds Conjugated with Carbon Dots 773.3.1.1. CDs Conjugated with Proteins/Peptides 783.3.1.2. CD Conjugates of Amino: Carboxylic Acid Ratio 803.3.1.3. CDs Conjugated with DNA 803.3.1.4. CDs Conjugated with RNase and SiRNA 843.3.1.5. CDs Conjugated with Lipid 863.3.1.6. CDs Conjugated with Folic Acid 863.3.1.7. CDs Conjugated with Chitosan 883.3.1.8. CDs Conjugated with Digitonin 893.3.2. Inorganic Heteroatoms Conjugated with CDs 903.3.2.1. CDs Conjugated with Gold Nanoparticles 913.3.2.2. CDs Conjugated with Silica 923.3.2.3. CDs Conjugated with ZnO 943.3.2.4. CDs Conjugated with CdS 953.3.2.5. CDs Conjugated with Strontium Oxide 963.3.2.6. CDs Conjugated with Gadolinium(III) 973.3.2.7. CDs Conjugated with Europium 973.3.2.8. CDs Conjugated/Doped with Nitrogen, Sulphur, Phosphorus and Boron 993.3.3. Carbon Dots Conjugated with Organic Material 1003.3.3.1. PEG (Polyethylene Glycol) 1013.3.3.2. CDs Conjugated with PEI (Polyethylenimin) or Polyaziridine 1023.3.3.3. CDs Conjugated with α-Cyclodextrin 1053.3.3.4. CDs Conjugated with Cysteamine 1063.3.3.5. CDs Conjugated with Dihydrolipoic Acid 1063.3.3.6. CDs Conjugated with Polyamidoamine (PAMAM) Dendrimers 1073.3.3.7. CDs Covalently Conjugated with Rhodamine B Dyes 1083.3.3.8. CDs Conjugated with Fe–Aminoclay (FeAC) 1093.3.3.9. CDs Conjugated with MWCNT 1093.3.4. CDs Conjugated with Antibiotics 1103.3.4.1. CDs Conjugated with Ciprofloxacin 1113.3.4.2. CDs Conjugated with Tetracycline 1143.3.4.3. CDs Conjugated with Vancomycin 1143.3.4.4. CDs Conjugated with Ampicillin 1153.3.4.5. CDs Conjugated with Streptomycin 1163.3.5. CDs Conjugated with Anti-Neurodegenerative Drugs for Delivery to Central Nervous System 1183.3.5.1. CDs Conjugated with Haloperidol 1193.3.5.2. CDs Conjugated with Transferrin 1243.3.5.3. CDs Conjugated with Curcumin 1253.3.6. CDs Conjugated with Anticancer Drugs 1283.3.6.1. CDs Conjugated with Doxorubicin 1283.3.6.2. CDs Conjugated with Cisplatin 1303.4. Summary 1324. CD as Drug Delivery Vehicle 1334.1. Introduction 1334.2. Considerations in Using CD as Drug Delivery Vehicle 1364.3. Designs of CD-Based Drug Delivery System 1374.3.1. Designing for Water-Insoluble Drugs 1384.3.2. Designing for Targeting Tumor Location 1384.3.3. Designing a Theranostic Nanomedicine 1394.3.4. Designing a Photoresponsive Nzzano Drug Delivery System 1394.3.5. Designing for Gene Delivery 1404.3.6. Designing for Antibiotics Delivery 1414.4. Carbon Dots for Delivery of Anti-Cancer Drug 1424.4.1. A Brief Introduction to Cancer 1434.4.2. Necessity of Drug Targeting in Cancer Therapy 1444.4.3. Targeting Angiogenesis with CD 1444.4.4. Various CD Conjugates for Delivering Anti-Cancer Drug 1454.4.5. CD for pH-Dependent Drug Release 1464.4.6. CD for Drug Delivery to Renal Cancer 1474.4.7. CD for Drug Delivery to Lung Cancer 1484.4.8. CD for Drug Delivery to Breast Cancer 1494.5. CD for Drug Delivery to Neurodegenerative Disease 1504.6. CD for Gene Therapy 1514.7. CD to Monitor Delivery of SiRNA 1524.8. Challenges in Using CD as Drug Delivery Vehicle 1524.8.1. Prevention of Drug from Biological Degradation 1534.8.2. Effective Targeting 1544.8.3. Patient Compliance 1554.8.4. Cost Effectiveness 1554.9. Suitability of CD-Conjugated Drugs 1564.9.1. For Oral Drug Delivery 1564.9.2. By Inhalation 1564.9.3. As Transdermal Drug Delivery 1574.9.4. As Injection 1574.10. Summary 1575. Carbon Dots for Cell Imaging and Diagnostics 1595.1. Introduction 1595.2. Bioimaging 1625.2.1. Bioimaging of Cancerous Cells 1665.2.1.1. HeLa Cells 1685.2.1.2. Human Breast Cancer MCF-7 Cells and Human Breast Tumor Cells MDA-MB-468 1705.2.1.3. B16F11 and HEK293 Cells 1715.2.1.4. Ehrlich Ascites Carcinoma (EAC) Cells 1735.2.1.5. Human U87 Cell 1735.2.1.6. MGC-803 Human Gastric Cancer Cells 1745.2.1.7. A549 Adenocarcinomic Human Alveolar (Lung) Basal Epithelial Cells 1755.2.1.8. Human Hepatocellular Carcinoma Cells 1755.2.1.9. Kidney Proximal Tubule Cell Line(LLC-PK1) 1765.2.1.10. C6 Glioma Cells 1775.2.2. Bioimaging of Nucleus 1785.2.3. Bioimaging of Virus 1805.2.4. Bioimaging of Bacteria 1815.2.5. Bioimaging of Drosophila melanogaster 1835.3. CDs as Sensor 1845.3.1. Intracellular Detection of Ions 1855.3.1.1. Detection of Ag Ions 1855.3.1.2. CD for Detection of Cu Ion 1875.3.1.3. Detection of Fe3+ Ions 1905.3.1.4. Detection of Hg2+ Ions 1915.3.2. Detection of Small Molecules 1925.3.2.1. Detection of Nitric Oxide 1935.3.2.2. Detection of Phosphate 1935.3.2.3. Detection of Reactive Oxygen Species 1945.3.2.4. Detection of H2S 1945.3.2.5. Detection of TNT 1955.3.2.6. Detection of Hydroquinone 1955.3.2.7. Detection of Surfactant 1965.3.2.8. Detection of Humidity 1975.3.3. Detection of Biological pH Value 1985.3.4. Detection of Nucleic Acid 2005.3.5. Detection of Vitamins 2015.3.6. Detection of Protein and Enzymes 2025.3.7. Detection of Glucose 2025.3.8. Detection of Cancerous Cells 2035.3.9. Detection of Dopamine Neurotransmitter 2045.4. Concluding Remarks 2066. Suitability of Carbon Dots as Payload for Plants 2096.1. Introduction 2096.2. Suitability of Carbon Dots as a Payload for Plant 2116.2.1. Cytotoxicity of Carbon Dots 2126.2.2. Carbon Dots and Plant Growth 2136.2.2.1. Wheat 2156.2.2.2. Green Beans 2156.2.3. Is Cell Wall a Barrier for Carbon Dot Internalization? 2166.2.4. Other Possible Routes for Entry of Carbon Dots 2196.3. Carbon Dots and Plant Fertilizer 2216.3.1. Nitrogen 2226.3.2. Phosphorus 2246.3.3. Potassium 2256.3.4. Micronutrients 2266.4. Need for Sensor to Detect 2276.4.1. Plant Nutrient Status 2286.4.2. Water 2296.4.3. Pathological Status of Plant 2316.4.4. Residual Pesticide, Herbicide or Insecticide in Plants 2326.5. Carbon Dots and Foliar Application of Drugs on Plants 2336.5.1. Cuticular Layer 2336.5.2. Cell Wall 2356.5.3. Plasma Membrane 2356.6. Carbon Dots as Trojan Horse to Penetrate Foliar Surface for Therapeutic Molecule Delivery 2366.6.1. CD for Delivery of Antibiotics to Plants 2376.7. Concluding Remarks 240References 241

Madhuri Sharon, (Retd Director Reliance Industries LTD) PhD from Leicester University UK, Post-Doctoral Research from Bolton Institute of Technology U.K., is currently Director of NSN Research Centre for Nanotechnology & Bionanotechnology and Managing Director of Monad Nanotech as well as Adjunct-Professor University of Mumbai & Professor-Emeritus JJT University. She has published more than 130 papers, 4 books and 11 patents. Her research focuses on the synthesis, biosynthesis and application of various nanomaterials (graphene oxide, carbon dots, carbon nanomaterials and nanometals, SPIONS) in drug-delivery. Ashmi Mewada holds a PhD degree from Nagoya Institute of Technology, Japan. She is currently an Assistant Professor in the Department of Nanotechnology at Walchand College of Arts and Science, Solapur. She has more than 30 publications and works extensively in the field of carbon dots.

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• Preface1. Carbon Dots: Discovery, Synthesis and Characterization 11.1. Background 11.2. Introduction to QD 21.2.1. What is Quantum Mechanics? 41.2.2. Quantum Confinement 71.2.3. Discovery and History of Carbon Dots 81.3. Carbon QD and Graphene QD 91.4. Various Methods of Synthesis of Carbon Dots 101.4.1. Electrochemical Methods 111.4.2. Combustion and Thermal Oxidation Method 131.4.3. Hydrothermal Oxidation Method 151.4.4. Solvothermal Method 181.4.5. Laser Ablation of Graphite 181.4.6. Pulsed Laser Irradiation of Carbon Source 201.4.7. Arc Discharge Method 201.4.8. Plasma Treatment 211.4.9. Opening of Fullerene Cage 221.4.10. Ultrasonication Method 221.4.11. Microwave-Assisted Method 231.4.12. Chemical Methods 261.4.13. Supported Synthetic Procedure 261.4.14. Biogenic Method 281.5. Characterization of Carbon Dots 311.5.1. Microscopic Methods 321.5.1.1. SEM and TEM Characterization 321.5.1.2. AFM and STM Characterization 341.5.2. Spectroscopic Methods 351.5.2.1. UV-Vis Spectroscopy and its Application for Band Gap Determination 371.5.2.2. Fluorescence Spectrometry 371.5.2.3. Fourier Transform Infrared (FTIR) Spectroscopy 381.5.2.4. X-Ray Diffraction (XRD) Analysis 401.5.2.5. X-Ray Photoelectron Spectroscopy (XPS) 411.5.2.6. Dynamic Light Scattering/Photon Correlation Spectroscopy (DLS/PCS) 411.5.2.7. Dual Polarization Interferometry (DPI) 421.5.2.8. Raman Spectroscopy 431.5.2.9. Nuclear Magnetic Resonance (NMR) Spectroscopy 441.6. Summary 452. Properties of Carbon Dots 472.1. Introduction 472.2. Optical Properties 492.2.1. Absorbance 512.2.2. Photo-Induced Electron Transfer (PET) with CDs 522.2.3. Fluorescence/Photoluminescence (PL) 532.2.3.1. Multiphoton Excitation 602.2.3.2. Upconversion Photoluminescence 612.2.3.3. Lack of Blinking 632.2.3.4. Resistance to Photobleaching 642.2.4. Photocatalytic Property 652.3. Chemically Inert 672.4. Easy Functionalization 672.5. Water Solubility 682.6. Low Toxicity 682.7. Biocompatibility 692.8. Summary 703. Carbon Dots and Conjugates 713.1. Introduction 713.2. Why Conjugation of Carbon Dots? 743.3. Types of Carbon Dot Conjugates and Their Applications 763.3.1. Biogenic Compounds Conjugated with Carbon Dots 773.3.1.1. CDs Conjugated with Proteins/Peptides 783.3.1.2. CD Conjugates of Amino: Carboxylic Acid Ratio 803.3.1.3. CDs Conjugated with DNA 803.3.1.4. CDs Conjugated with RNase and SiRNA 843.3.1.5. CDs Conjugated with Lipid 863.3.1.6. CDs Conjugated with Folic Acid 863.3.1.7. CDs Conjugated with Chitosan 883.3.1.8. CDs Conjugated with Digitonin 893.3.2. Inorganic Heteroatoms Conjugated with CDs 903.3.2.1. CDs Conjugated with Gold Nanoparticles 913.3.2.2. CDs Conjugated with Silica 923.3.2.3. CDs Conjugated with ZnO 943.3.2.4. CDs Conjugated with CdS 953.3.2.5. CDs Conjugated with Strontium Oxide 963.3.2.6. CDs Conjugated with Gadolinium(III) 973.3.2.7. CDs Conjugated with Europium 973.3.2.8. CDs Conjugated/Doped with Nitrogen, Sulphur, Phosphorus and Boron 993.3.3. Carbon Dots Conjugated with Organic Material 1003.3.3.1. PEG (Polyethylene Glycol) 1013.3.3.2. CDs Conjugated with PEI (Polyethylenimin) or Polyaziridine 1023.3.3.3. CDs Conjugated with α-Cyclodextrin 1053.3.3.4. CDs Conjugated with Cysteamine 1063.3.3.5. CDs Conjugated with Dihydrolipoic Acid 1063.3.3.6. CDs Conjugated with Polyamidoamine (PAMAM) Dendrimers 1073.3.3.7. CDs Covalently Conjugated with Rhodamine B Dyes 1083.3.3.8. CDs Conjugated with Fe–Aminoclay (FeAC) 1093.3.3.9. CDs Conjugated with MWCNT 1093.3.4. CDs Conjugated with Antibiotics 1103.3.4.1. CDs Conjugated with Ciprofloxacin 1113.3.4.2. CDs Conjugated with Tetracycline 1143.3.4.3. CDs Conjugated with Vancomycin 1143.3.4.4. CDs Conjugated with Ampicillin 1153.3.4.5. CDs Conjugated with Streptomycin 1163.3.5. CDs Conjugated with Anti-Neurodegenerative Drugs for Delivery to Central Nervous System 1183.3.5.1. CDs Conjugated with Haloperidol 1193.3.5.2. CDs Conjugated with Transferrin 1243.3.5.3. CDs Conjugated with Curcumin 1253.3.6. CDs Conjugated with Anticancer Drugs 1283.3.6.1. CDs Conjugated with Doxorubicin 1283.3.6.2. CDs Conjugated with Cisplatin 1303.4. Summary 1324. CD as Drug Delivery Vehicle 1334.1. Introduction 1334.2. Considerations in Using CD as Drug Delivery Vehicle 1364.3. Designs of CD-Based Drug Delivery System 1374.3.1. Designing for Water-Insoluble Drugs 1384.3.2. Designing for Targeting Tumor Location 1384.3.3. Designing a Theranostic Nanomedicine 1394.3.4. Designing a Photoresponsive Nzzano Drug Delivery System 1394.3.5. Designing for Gene Delivery 1404.3.6. Designing for Antibiotics Delivery 1414.4. Carbon Dots for Delivery of Anti-Cancer Drug 1424.4.1. A Brief Introduction to Cancer 1434.4.2. Necessity of Drug Targeting in Cancer Therapy 1444.4.3. Targeting Angiogenesis with CD 1444.4.4. Various CD Conjugates for Delivering Anti-Cancer Drug 1454.4.5. CD for pH-Dependent Drug Release 1464.4.6. CD for Drug Delivery to Renal Cancer 1474.4.7. CD for Drug Delivery to Lung Cancer 1484.4.8. CD for Drug Delivery to Breast Cancer 1494.5. CD for Drug Delivery to Neurodegenerative Disease 1504.6. CD for Gene Therapy 1514.7. CD to Monitor Delivery of SiRNA 1524.8. Challenges in Using CD as Drug Delivery Vehicle 1524.8.1. Prevention of Drug from Biological Degradation 1534.8.2. Effective Targeting 1544.8.3. Patient Compliance 1554.8.4. Cost Effectiveness 1554.9. Suitability of CD-Conjugated Drugs 1564.9.1. For Oral Drug Delivery 1564.9.2. By Inhalation 1564.9.3. As Transdermal Drug Delivery 1574.9.4. As Injection 1574.10. Summary 1575. Carbon Dots for Cell Imaging and Diagnostics 1595.1. Introduction 1595.2. Bioimaging 1625.2.1. Bioimaging of Cancerous Cells 1665.2.1.1. HeLa Cells 1685.2.1.2. Human Breast Cancer MCF-7 Cells and Human Breast Tumor Cells MDA-MB-468 1705.2.1.3. B16F11 and HEK293 Cells 1715.2.1.4. Ehrlich Ascites Carcinoma (EAC) Cells 1735.2.1.5. Human U87 Cell 1735.2.1.6. MGC-803 Human Gastric Cancer Cells 1745.2.1.7. A549 Adenocarcinomic Human Alveolar (Lung) Basal Epithelial Cells 1755.2.1.8. Human Hepatocellular Carcinoma Cells 1755.2.1.9. Kidney Proximal Tubule Cell Line(LLC-PK1) 1765.2.1.10. C6 Glioma Cells 1775.2.2. Bioimaging of Nucleus 1785.2.3. Bioimaging of Virus 1805.2.4. Bioimaging of Bacteria 1815.2.5. Bioimaging of Drosophila melanogaster 1835.3. CDs as Sensor 1845.3.1. Intracellular Detection of Ions 1855.3.1.1. Detection of Ag Ions 1855.3.1.2. CD for Detection of Cu Ion 1875.3.1.3. Detection of Fe3+ Ions 1905.3.1.4. Detection of Hg2+ Ions 1915.3.2. Detection of Small Molecules 1925.3.2.1. Detection of Nitric Oxide 1935.3.2.2. Detection of Phosphate 1935.3.2.3. Detection of Reactive Oxygen Species 1945.3.2.4. Detection of H2S 1945.3.2.5. Detection of TNT 1955.3.2.6. Detection of Hydroquinone 1955.3.2.7. Detection of Surfactant 1965.3.2.8. Detection of Humidity 1975.3.3. Detection of Biological pH Value 1985.3.4. Detection of Nucleic Acid 2005.3.5. Detection of Vitamins 2015.3.6. Detection of Protein and Enzymes 2025.3.7. Detection of Glucose 2025.3.8. Detection of Cancerous Cells 2035.3.9. Detection of Dopamine Neurotransmitter 2045.4. Concluding Remarks 2066. Suitability of Carbon Dots as Payload for Plants 2096.1. Introduction 2096.2. Suitability of Carbon Dots as a Payload for Plant 2116.2.1. Cytotoxicity of Carbon Dots 2126.2.2. Carbon Dots and Plant Growth 2136.2.2.1. Wheat 2156.2.2.2. Green Beans 2156.2.3. Is Cell Wall a Barrier for Carbon Dot Internalization? 2166.2.4. Other Possible Routes for Entry of Carbon Dots 2196.3. Carbon Dots and Plant Fertilizer 2216.3.1. Nitrogen 2226.3.2. Phosphorus 2246.3.3. Potassium 2256.3.4. Micronutrients 2266.4. Need for Sensor to Detect 2276.4.1. Plant Nutrient Status 2286.4.2. Water 2296.4.3. Pathological Status of Plant 2316.4.4. Residual Pesticide, Herbicide or Insecticide in Plants 2326.5. Carbon Dots and Foliar Application of Drugs on Plants 2336.5.1. Cuticular Layer 2336.5.2. Cell Wall 2356.5.3. Plasma Membrane 2356.6. Carbon Dots as Trojan Horse to Penetrate Foliar Surface for Therapeutic Molecule Delivery 2366.6.1. CD for Delivery of Antibiotics to Plants 2376.7. Concluding Remarks 240References 241

Madhuri Sharon, (Retd Director Reliance Industries LTD) PhD from Leicester University UK, Post-Doctoral Research from Bolton Institute of Technology U.K., is currently Director of NSN Research Centre for Nanotechnology & Bionanotechnology and Managing Director of Monad Nanotech as well as Adjunct-Professor University of Mumbai & Professor-Emeritus JJT University. She has published more than 130 papers, 4 books and 11 patents. Her research focuses on the synthesis, biosynthesis and application of various nanomaterials (graphene oxide, carbon dots, carbon nanomaterials and nanometals, SPIONS) in drug-delivery. Ashmi Mewada holds a PhD degree from Nagoya Institute of Technology, Japan. She is currently an Assistant Professor in the Department of Nanotechnology at Walchand College of Arts and Science, Solapur. She has more than 30 publications and works extensively in the field of carbon dots.

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• Preface1. Carbon Dots: Discovery, Synthesis and Characterization 11.1. Background 11.2. Introduction to QD 21.2.1. What is Quantum Mechanics? 41.2.2. Quantum Confinement 71.2.3. Discovery and History of Carbon Dots 81.3. Carbon QD and Graphene QD 91.4. Various Methods of Synthesis of Carbon Dots 101.4.1. Electrochemical Methods 111.4.2. Combustion and Thermal Oxidation Method 131.4.3. Hydrothermal Oxidation Method 151.4.4. Solvothermal Method 181.4.5. Laser Ablation of Graphite 181.4.6. Pulsed Laser Irradiation of Carbon Source 201.4.7. Arc Discharge Method 201.4.8. Plasma Treatment 211.4.9. Opening of Fullerene Cage 221.4.10. Ultrasonication Method 221.4.11. Microwave-Assisted Method 231.4.12. Chemical Methods 261.4.13. Supported Synthetic Procedure 261.4.14. Biogenic Method 281.5. Characterization of Carbon Dots 311.5.1. Microscopic Methods 321.5.1.1. SEM and TEM Characterization 321.5.1.2. AFM and STM Characterization 341.5.2. Spectroscopic Methods 351.5.2.1. UV-Vis Spectroscopy and its Application for Band Gap Determination 371.5.2.2. Fluorescence Spectrometry 371.5.2.3. Fourier Transform Infrared (FTIR) Spectroscopy 381.5.2.4. X-Ray Diffraction (XRD) Analysis 401.5.2.5. X-Ray Photoelectron Spectroscopy (XPS) 411.5.2.6. Dynamic Light Scattering/Photon Correlation Spectroscopy (DLS/PCS) 411.5.2.7. Dual Polarization Interferometry (DPI) 421.5.2.8. Raman Spectroscopy 431.5.2.9. Nuclear Magnetic Resonance (NMR) Spectroscopy 441.6. Summary 452. Properties of Carbon Dots 472.1. Introduction 472.2. Optical Properties 492.2.1. Absorbance 512.2.2. Photo-Induced Electron Transfer (PET) with CDs 522.2.3. Fluorescence/Photoluminescence (PL) 532.2.3.1. Multiphoton Excitation 602.2.3.2. Upconversion Photoluminescence 612.2.3.3. Lack of Blinking 632.2.3.4. Resistance to Photobleaching 642.2.4. Photocatalytic Property 652.3. Chemically Inert 672.4. Easy Functionalization 672.5. Water Solubility 682.6. Low Toxicity 682.7. Biocompatibility 692.8. Summary 703. Carbon Dots and Conjugates 713.1. Introduction 713.2. Why Conjugation of Carbon Dots? 743.3. Types of Carbon Dot Conjugates and Their Applications 763.3.1. Biogenic Compounds Conjugated with Carbon Dots 773.3.1.1. CDs Conjugated with Proteins/Peptides 783.3.1.2. CD Conjugates of Amino: Carboxylic Acid Ratio 803.3.1.3. CDs Conjugated with DNA 803.3.1.4. CDs Conjugated with RNase and SiRNA 843.3.1.5. CDs Conjugated with Lipid 863.3.1.6. CDs Conjugated with Folic Acid 863.3.1.7. CDs Conjugated with Chitosan 883.3.1.8. CDs Conjugated with Digitonin 893.3.2. Inorganic Heteroatoms Conjugated with CDs 903.3.2.1. CDs Conjugated with Gold Nanoparticles 913.3.2.2. CDs Conjugated with Silica 923.3.2.3. CDs Conjugated with ZnO 943.3.2.4. CDs Conjugated with CdS 953.3.2.5. CDs Conjugated with Strontium Oxide 963.3.2.6. CDs Conjugated with Gadolinium(III) 973.3.2.7. CDs Conjugated with Europium 973.3.2.8. CDs Conjugated/Doped with Nitrogen, Sulphur, Phosphorus and Boron 993.3.3. Carbon Dots Conjugated with Organic Material 1003.3.3.1. PEG (Polyethylene Glycol) 1013.3.3.2. CDs Conjugated with PEI (Polyethylenimin) or Polyaziridine 1023.3.3.3. CDs Conjugated with α-Cyclodextrin 1053.3.3.4. CDs Conjugated with Cysteamine 1063.3.3.5. CDs Conjugated with Dihydrolipoic Acid 1063.3.3.6. CDs Conjugated with Polyamidoamine (PAMAM) Dendrimers 1073.3.3.7. CDs Covalently Conjugated with Rhodamine B Dyes 1083.3.3.8. CDs Conjugated with Fe–Aminoclay (FeAC) 1093.3.3.9. CDs Conjugated with MWCNT 1093.3.4. CDs Conjugated with Antibiotics 1103.3.4.1. CDs Conjugated with Ciprofloxacin 1113.3.4.2. CDs Conjugated with Tetracycline 1143.3.4.3. CDs Conjugated with Vancomycin 1143.3.4.4. CDs Conjugated with Ampicillin 1153.3.4.5. CDs Conjugated with Streptomycin 1163.3.5. CDs Conjugated with Anti-Neurodegenerative Drugs for Delivery to Central Nervous System 1183.3.5.1. CDs Conjugated with Haloperidol 1193.3.5.2. CDs Conjugated with Transferrin 1243.3.5.3. CDs Conjugated with Curcumin 1253.3.6. CDs Conjugated with Anticancer Drugs 1283.3.6.1. CDs Conjugated with Doxorubicin 1283.3.6.2. CDs Conjugated with Cisplatin 1303.4. Summary 1324. CD as Drug Delivery Vehicle 1334.1. Introduction 1334.2. Considerations in Using CD as Drug Delivery Vehicle 1364.3. Designs of CD-Based Drug Delivery System 1374.3.1. Designing for Water-Insoluble Drugs 1384.3.2. Designing for Targeting Tumor Location 1384.3.3. Designing a Theranostic Nanomedicine 1394.3.4. Designing a Photoresponsive Nzzano Drug Delivery System 1394.3.5. Designing for Gene Delivery 1404.3.6. Designing for Antibiotics Delivery 1414.4. Carbon Dots for Delivery of Anti-Cancer Drug 1424.4.1. A Brief Introduction to Cancer 1434.4.2. Necessity of Drug Targeting in Cancer Therapy 1444.4.3. Targeting Angiogenesis with CD 1444.4.4. Various CD Conjugates for Delivering Anti-Cancer Drug 1454.4.5. CD for pH-Dependent Drug Release 1464.4.6. CD for Drug Delivery to Renal Cancer 1474.4.7. CD for Drug Delivery to Lung Cancer 1484.4.8. CD for Drug Delivery to Breast Cancer 1494.5. CD for Drug Delivery to Neurodegenerative Disease 1504.6. CD for Gene Therapy 1514.7. CD to Monitor Delivery of SiRNA 1524.8. Challenges in Using CD as Drug Delivery Vehicle 1524.8.1. Prevention of Drug from Biological Degradation 1534.8.2. Effective Targeting 1544.8.3. Patient Compliance 1554.8.4. Cost Effectiveness 1554.9. Suitability of CD-Conjugated Drugs 1564.9.1. For Oral Drug Delivery 1564.9.2. By Inhalation 1564.9.3. As Transdermal Drug Delivery 1574.9.4. As Injection 1574.10. Summary 1575. Carbon Dots for Cell Imaging and Diagnostics 1595.1. Introduction 1595.2. Bioimaging 1625.2.1. Bioimaging of Cancerous Cells 1665.2.1.1. HeLa Cells 1685.2.1.2. Human Breast Cancer MCF-7 Cells and Human Breast Tumor Cells MDA-MB-468 1705.2.1.3. B16F11 and HEK293 Cells 1715.2.1.4. Ehrlich Ascites Carcinoma (EAC) Cells 1735.2.1.5. Human U87 Cell 1735.2.1.6. MGC-803 Human Gastric Cancer Cells 1745.2.1.7. A549 Adenocarcinomic Human Alveolar (Lung) Basal Epithelial Cells 1755.2.1.8. Human Hepatocellular Carcinoma Cells 1755.2.1.9. Kidney Proximal Tubule Cell Line(LLC-PK1) 1765.2.1.10. C6 Glioma Cells 1775.2.2. Bioimaging of Nucleus 1785.2.3. Bioimaging of Virus 1805.2.4. Bioimaging of Bacteria 1815.2.5. Bioimaging of Drosophila melanogaster 1835.3. CDs as Sensor 1845.3.1. Intracellular Detection of Ions 1855.3.1.1. Detection of Ag Ions 1855.3.1.2. CD for Detection of Cu Ion 1875.3.1.3. Detection of Fe3+ Ions 1905.3.1.4. Detection of Hg2+ Ions 1915.3.2. Detection of Small Molecules 1925.3.2.1. Detection of Nitric Oxide 1935.3.2.2. Detection of Phosphate 1935.3.2.3. Detection of Reactive Oxygen Species 1945.3.2.4. Detection of H2S 1945.3.2.5. Detection of TNT 1955.3.2.6. Detection of Hydroquinone 1955.3.2.7. Detection of Surfactant 1965.3.2.8. Detection of Humidity 1975.3.3. Detection of Biological pH Value 1985.3.4. Detection of Nucleic Acid 2005.3.5. Detection of Vitamins 2015.3.6. Detection of Protein and Enzymes 2025.3.7. Detection of Glucose 2025.3.8. Detection of Cancerous Cells 2035.3.9. Detection of Dopamine Neurotransmitter 2045.4. Concluding Remarks 2066. Suitability of Carbon Dots as Payload for Plants 2096.1. Introduction 2096.2. Suitability of Carbon Dots as a Payload for Plant 2116.2.1. Cytotoxicity of Carbon Dots 2126.2.2. Carbon Dots and Plant Growth 2136.2.2.1. Wheat 2156.2.2.2. Green Beans 2156.2.3. Is Cell Wall a Barrier for Carbon Dot Internalization? 2166.2.4. Other Possible Routes for Entry of Carbon Dots 2196.3. Carbon Dots and Plant Fertilizer 2216.3.1. Nitrogen 2226.3.2. Phosphorus 2246.3.3. Potassium 2256.3.4. Micronutrients 2266.4. Need for Sensor to Detect 2276.4.1. Plant Nutrient Status 2286.4.2. Water 2296.4.3. Pathological Status of Plant 2316.4.4. Residual Pesticide, Herbicide or Insecticide in Plants 2326.5. Carbon Dots and Foliar Application of Drugs on Plants 2336.5.1. Cuticular Layer 2336.5.2. Cell Wall 2356.5.3. Plasma Membrane 2356.6. Carbon Dots as Trojan Horse to Penetrate Foliar Surface for Therapeutic Molecule Delivery 2366.6.1. CD for Delivery of Antibiotics to Plants 2376.7. Concluding Remarks 240References 241