Molecular Fluorescent Sensors for Cellular StudiesEnables readers to fully understand the fundamentals and chemical principles of fluorescent sensing and the design of fluorescent sensors
Fluorescent sensors are able to provide specific chemical information about cells and can be invaluable in understanding processes that underpin health and disease.Molecular Fluorescent Sensors for Cellular Studies provides an avenue into and overview of currently available fluorescent sensing technology and its application to biological imaging.
This book aims to help the reader understand the principles of fluorescence and the mechanisms by which fluorescent sensors operate in order to ensure appropriate and optimal use of sensors. Key applications of fluorescent sensing are presented, with explanations not only of how new sensors can be designed, but also how existing sensors can be applied to various biological settings and conditions. Clear and engaging schematics throughout the book explain chemical principles of sensing to the non-expert.
Discusses the breadth of fluorescent sensors, from commercially available sensors to those reported in literature which are yet to be used widely Explains how fluorescent sensors operate for appropriate and optimal use from a theoretical standpoint Provides guidance on how to achieve optimal use of fluorescent sensors in practical settings Summarizes the principles behind fluorescent sensors and their design
This work will be an invaluable resource for postgraduates and professionals in the fields of microscopy, bioimaging, and diagnostic imaging who wish to harness the information to improve practical applications and to gain key knowledge surrounding the many facets of fluorescent sensing. It is also of interest to medical and biological researchers working across industry, universities and medical institutes.
Author(s): Elizabeth J. New
Publisher: Wiley
Year: 2022
Language: English
Pages: 306
City: Hoboken
Cover
Title Page
Copyright Page
Contents
List of Contributors
Chapter 1 An Introduction to Small Molecule Fluorescent Sensors
1.1 What is Fluorescence?
1.2 Why Is Fluorescence Useful?
1.3 What Is a Fluorescent Sensor?
1.4 General Types of Fluorescent Sensors
1.5 Important Parameters
1.5.1 Excitation Maxima
1.5.2 Emission Maxima
1.5.3 Stokes Shift
1.5.4 Quantum Yield
1.5.5 Molar Extinction Coefficient
1.5.6 Brightness
1.5.7 Lifetime
1.5.8 Photobleaching
1.5.9 Anisotropy
1.5.10 Quenching
1.6 Fluorescence Mechanisms Used in Fluorescent Sensors
1.6.1 Photoinduced Electron Transfer
1.6.2 Internal Charge Transfer
1.6.3 Förster Resonance Energy Transfer
1.6.4 Through Bond Energy Transfer
1.6.5 Excited-State Intramolecular Proton Transfer
1.6.6 Aggregation-Induced Emission
1.6.7 Excimer Formation
1.7 Commonly Used Fluorophores
1.7.1 Fluorescein
1.7.2 Rhodamine
1.7.3 Coumarin
1.7.4 Naphthalimide
1.7.5 BODIPY (4,4-Difluoro4-bora-3a,4a-diaza-s-indacene)
1.7.6 Cyanine
1.8 Summary
References
Chapter 2 The Applications of Responsive Fluorescent Sensors to Biological Systems
2.1 Criteria for Biologically Relevant Fluorescent Sensors
2.2 Microscopy for Visualising Fluorescent Sensors
2.2.1 Important Considerations in Microscopy
2.2.2 Common Microscopy Techniques
2.3 Other Instrumental Techniques for Studying Cells Treated with Fluorescent Sensors
2.3.1 Flow Cytometry
2.3.2 Fluorescence Plate-readers
2.4 Biological Samples to Which Fluorescent Sensors Can Be Applied
2.4.1 Cultured Mammalian Cells
2.4.2 Bacteria
2.4.3 Plants
2.4.4 Multi-cellular Organisms
2.4.5 Towards In Vivo Imaging
2.5 Common Challenges and Misconceptions in the Applications of Fluorescent Sensors
2.5.1 Important Considerations in Applying Sensors
2.5.2 Common Misconceptions About the Use of Sensors – The Bridge Between Multiple Disciplines
2.6 Conclusions
References
Chapter 3 Methods to Control the Subcellular Localisation of Fluorescent Sensors
3.1 Introduction
3.2 Targeting the Nucleus
3.3 Targeting Mitochondria
3.4 Targeting Lysosomes
3.5 Targeting Endosomes
3.6 Targeting Autophagic Compartments
3.7 Targeting Peroxisomes
3.8 Targeting the Endoplasmic Reticulum
3.9 Targeting the Golgi Apparatus
3.10 Targeting Lipid Droplets
3.11 Targeting the Plasma Membrane
3.12 Targeting the Cytoskeleton
3.13 Targeting the Cytosol
3.14 Trapping and Accumulation of Sensors
References
Chapter 4 Recognition-based Sensors for Cellular Imaging
4.1 Considerations for Recognition-based Sensing
4.1.1 Receptor–Analyte Recognition and Binding Affinity
4.1.2 Key Considerations to Enhance Selective Receptor to Analyte Recognition
4.2 Recognition-based Cation Sensing
4.2.1 Group I and II Metal Sensing
4.2.2 Essential Transition Metal Sensing
4.2.3 Toxic Metal Sensing
4.3 Recognition-based Anion Sensing
4.3.1 Anion Sensing Approaches
4.3.2 Halogen Ions Sensing
4.3.3 Inorganic Phosphates and Pyrophosphates
4.3.4 Bicarbonate, Hydrogen Sulfate, and Nitrate
4.4 Conclusions
References
Chapter 5 Activity-based Fluorescent Sensors and Their Applications in Biological Studies
5.1 Introduction
5.1.1 Design Principles
5.2 Oxidation Reactions for Sensing Oxidative Species
5.2.1 Fluorescent Sensors for Hydrogen Peroxide
5.2.2 Fluorescent Sensors for Peroxynitrite
5.2.3 Fluorescent Sensors for Hypochlorous Acid
5.2.4 Fluorescent Sensors for Nitric Oxide
5.2.5 Fluorescent Sensors for Singlet Oxygen
5.3 Reduction Reactions for Sensing Reductive Species
5.3.1 Fluorescent Sensors for Hydrogen Sulfides
5.3.2 Fluorescent Sensors for Glutathione, Cysteine, and Homocysteine
5.3.3 Fluorescent Sensors for Selenocysteine
5.4 Reactions for Sensing Carbonyl Species
5.4.1 Fluorescent Sensors for Formaldehyde
5.4.2 Fluorescent Sensors for Methylglyoxal
5.5 Metal-mediated Reactions
5.6 Metal-sensing Reactions
5.7 Enzymatic Reactions
5.8 Reversible Reactions
5.8.1 Nucleophilic Conjugate Additions
5.8.2 Nucleophilic Addition
5.8.3 Imine Formation
5.8.4 Oxidation–Reduction Reactions
5.9 Analyte Regeneration
5.10 Summary
References
Chapter 6 Fluorescent Sensors of the Cellular Environment
6.1 Fluorescent Sensors for Polarity and Viscosity
6.1.1 The Biological Significance of Polarity and Viscosity
6.1.2 Twisted Intramolecular Charge Transfer as a Mechanism for Polarity and Viscosity Sensing
6.1.3 Polarity Sensors Based on Other Mechanisms
6.2 Fluorescent Sensors for pH
6.2.1 The Regulation of pH in Health and Disease
6.2.2 Considerations and Design Strategies for the Preparation of pH Sensors
6.2.3 Examples of pH Sensors
6.3 Fluorescent Redox Sensors for Biological Studies
6.3.1 The Regulation of Redox State in Health and Disease
6.3.2 Design Strategies of Fluorescent Redox Sensors and Key Examples
6.4 Conclusions
References
Chapter 7 Labelling Proteins and Biomolecules with Small Fluorescent Sensors
7.1 Labelling Biomolecules in Cells with Fluorescent Sensors
7.2 Small-molecule Modifications and Bioorthogonal Reactions
7.2.1 Polar Ketone and Aldehyde Condensations
7.2.2 Azide Bioorthogonal Chemistry
7.2.3 Tetrazine Ligation
7.2.4 Commercial Fluorescent Labels
7.3 Short peptide Recognition Sequences
7.4 Fusion Protein Tagging Systems
7.4.1 FKBP Tag
7.4.2 eDHFR Tag
7.4.3 PYP Tag
7.4.4 SNAP-Tag and CLIP-Tag
7.4.5 HaloTag
7.5 Enzymatic Modifications for Labelling Proteins
7.5.1 The LAP-tag System
7.5.2 Protein Trans-splicing
7.6 Future Developments
References
Chapter 8 Future Directions of Fluorescence Sensors for Cellular Studies
8.1 Fluorescence Lifetime Imaging Microscopy
8.1.1 Introduction
8.1.2 Advantages of Fluorescence Lifetime Imaging Microscopy
8.1.3 Examples of Sensors for FLIM
8.1.4 Future Directions
8.2 Near-infrared Sensors
8.2.1 Strategies to Make NIR Sensors
8.2.2 NIR Fluorophore Scaffolds
8.2.3 Future Directions
8.3 Dual-analyte Sensing
8.3.1 Introduction
8.3.2 Recognition-Based Dual-analyte Sensors
8.3.3 Activity-based Dual-analyte Sensors
8.3.4 Mixed Dual-analyte Sensors
8.3.5 Sequence-specific Reactions
8.3.6 Conclusions and Future Directions
8.4 Super-resolution Microscopy
8.4.1 Introduction
8.4.2 Super-resolution Microscopy Techniques
8.4.3 Considerations for Use of Super Resolution Microscopy
8.4.4 Fluorescent Sensors for Super-resolution Microscopy
8.4.5 Future Directions
8.5 Multimodality
8.5.1 Introduction
8.5.2 Radioisotope Techniques
8.5.3 Computed Tomography
8.5.4 Magnetic Resonance Imaging
8.5.5 Photoacoustic Imaging
8.5.6 Vibrational Spectroscopy
8.5.7 Synchrotron X-ray Techniques
8.5.8 Mass Spectrometry
8.5.9 Electron Microscopy
8.5.10 Three or More Modalities
8.5.11 Future Directions
References
Index
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