Fluorescence-Based Biosensors, Volume 113

Contributors

Preface

Introduction

Chapter One. The Chemistry of Small-Molecule Fluorogenic Probes

1 Introduction

2 Fluorescence and Fluorophores

3 Modes of Fluorescence Modulation

4 Coumarins

5 Bodipy Dyes

6 Fluoresceins

7 Rhodamines

8 Phenoxazine Dyes

9 Acridinones

10 Cyanines

11 Other Fluorogenic Scaffolds

12 Conclusions and Future Directions

References

Chapter Two. Fluorescent Environment-Sensitive Dyes as Reporters of Biomolecular Interactions

1 Introduction

2 Single-Band Solvatochromic Dyes: Design and Applications

3 Two-Band Solvatochromic Dyes Based on Esipt

4 Applications of Two-Color Dyes for Monitoring Biomolecular Interactions

5 Conclusions

References

Chapter Three. Rational Design of Fluorophores for In Vivo Applications

1 Introduction

2 Cyanine and Related Fluorophores

3 Squaraines

4 Boron Dipyrromethene and Related Fluorophores

5 Porphyrins, Phthalocyanines, and Related Macrocycles

6 Special Types of Fluorophores

7 Conclusion

References

Chapter Four. Fluorescence Technologies for Monitoring Interactions Between Biological Molecules In Vitro

Abbreviations

1 Introduction to Fluorescence

2 Monitoring Protein/Substrate Interactions

3 Monitoring Protein/Protein and Polypeptide Interactions

4 Fluorescence for Protein/Membrane Interactions

5 Conclusions and Perspectives

References

Chapter Five. From FRET Imaging to Practical Methodology for Kinase Activity Sensing in Living Cells

Abbreviations

1 Introduction

2 Fluorescence Generalities

3 FRET Measurement

4 FRET Measurements: Methods and Instrumentation

5 Data Analysis

6 Design and Optimization of Genetically Encoded KARs

7 Considerations for KAR Measurements

8 Toward Quantitative Approaches in Biological Processes

9 Outlook and Perspective

References

Chapter Six. Fluorescent Sensors of Protein Kinases

Abbreviations

1 Introduction

2 Genetically Encoded Reporters of Protein Kinases

3 Fluorescent Peptide/Protein Biosensors

4 Applications

5 Conclusions and Perspectives

References

Chapter Seven. Time-Resolved Förster Resonance Energy Transfer-Based Technologies to Investigate G Protein-Coupled Receptor Machinery

1 Introduction

2 Overview of the TR-FRET Principle and Its Advantages

3 Screening with Generic Methods

4 Screening with Target-Specific Methods

5 Perspectives

6 Conclusion

References

Chapter Eight. Fluorescent Protein-Based Biosensors and Their Clinical Applications

1 Introduction

2 Fluorescent Proteins

3 Single-Fluorescent Protein-Based Biosensors

4 Bimolecular Fluorescence Complementation

5 Förster Resonance Energy Transfer

6 Methods for Evaluating FRET Efficiency

7 Clinical Application of FRET

8 Closing Remarks

References

Chapter Nine. Fluorescent Macromolecular Sensors of Enzymatic Activity for In Vivo Imaging

1 Introduction

2 Enzyme Activity and Dysregulation as a Predictor of Disease

3 Fluorescence Detection In Vivo

4 Principles of Enzymatic Activity Sensors Based on Fluorescence Changes

5 Macromolecular Fluorescent Probes

6 Pharmacokinetics of PGC and Imaging of Activation

7 Applications of Macromolecular Fluorescent Sensors in Cancer Imaging

8 Alternative Enzyme-Targeted Strategies in Imaging Cancer

9 Macromolecular Sensors in Fluorescent Imaging of Inflammation and Vascular Disease

10 Conclusions

References

Chapter Ten. Fluorescent Proteins as Visible In Vivo Sensors

1 Introduction

2 Noninvasive Imaging

3 Lighting Up the Tumor Stroma with Fluorescent Proteins

4 Stroma Cells are Required for Cancer Metastasis

5 Fluorescent Tumorgrafts made from Human Cancer Patients

6 Real-Time Imaging of Trafficking Cancer Cells

7 Method of Choice for Whole-Body Imaging

8 Conclusions

References

Index

One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors.

Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed and engineered thanks to the combined efforts of chemists and biologists over the last decade, and developed hand in hand together with imaging technologies.

This volume will convey the many exciting developments the field of fluorescent biosensors and reporters has witnessed over the recent years, from concepts to applications, including chapters on the chemistry of fluorescent probes, on technologies for monitoring protein/protein interactions and technologies for imaging biosensors in cultured cells and in vivo. Other chapters are devoted to specific examples of genetically-encoded reporters, or to protein and peptide biosensors, together with examples illustrating their application to cellular and in vivo imaging, biomedical applications, drug discovery and high throughput screening.

• Contributions from leading authorities
• Informs and updates on all the latest developments in the field

Researchers in imaging and fluorescence or biosensors and bioelectronics