Fluorescence applications in biotechnology and life sciences / edited by Ewa M. Goldys. — Oxford : Wiley-Blackwell, c2009. – (58.1054/F646) |
Contents
CONTENTS
Preface
Acknowledgments
About the Contributing Authors
1 Basics of Fluorescence
1.1 Introduction
1.2 Absorption and Emission of Light
1.3 Nonradiative Decay Mechanisms
1.4 Properties of Excited Molecules
1.5 Spectroscopy and Fluorophores
1.6 Environmental Sensitivity of Fluorophores
1.7 Polarization of Fluorescence
1.8 Conclusion
References
2 Labeling of Cells with Fluorescent Dyes
2.1 Introduction
2.2 Fluorophore Selection
2.3 Loading and Labeling Live Cells
2.4 Fluorophores for Live Cell Imaging
References
3 Genetically Encoded Fluorescent Probes: Some Properties and Applications in Life Sciences
3.1 Introduction
3.2 Chromophore and its Formation
3.3 "Life and Death" of Fluorescent Protein
3.4 Applications
3.5 Passive Applications
3.6 Active Applications
3.7 Interactive Applications
3.8 Conclusions
References
4 Nanoparticle Fluorescence Probes
4.1 Introduction
4.2 Nanomaterials for Biological Applications
4.3 Inorganic Quantum Dots: Physics and Optical Properties
4.4 Synthesis of Monodisperse Colloidal Quantum Dots for Biolabeling Applications
4.5 Quantum Dots as In Vitro Probes
4.6 Quantum Dots as In Vivo Probes Cytotoxicity Future Directions References
5 Quantitative Analysis of Fluorescent Image: From Descriptive to Computational Microscopy
5.1 Introduction
5.2 Advantages of Quantitative Analysis
5.3 Methods of Quantitative Analysis
5.4 Image Processing
References
6 Spectral Imaging and Unmixing
6.1 Introduction
6.2 Instrumentation and Configurations for Hyperspectral Microscopes
6.3 Supervised and Unsupervised Unmixing
6.4 Supervised or Informed Unmixing
6.5 Unsupervised or Blind Unmixing
6.6 Spectral Clustering
6.7 Examples
6.8 Limitations of Spectral Imaging and Experimental Considerations
6.9 Conclusions
References
7 Correlation of Light with Electron Microscopy: A Correlative Microscopy Platform
7.1 Introduction
7.2 Overview of Techniques Used in Correlative Microscopy
7.3 Correlative Microscopy of Chemically Fixed, Immunolabeled Ultrathin Cryosections Employing Antibodies Coupled with Fluorescent Gold Conjugates
7.4 Special Techniques Used in Sample Preparation for Correlative Microscopy: High-Pressure Freezing, Freeze Fracturing, and Freeze Substitution
7.5 Correlative Microscopy of Fixed Tissue Specimens Using Freeze Substitution
7.6 Future Trends: Correlative Microscopy and High-Content Cellular Screening
7.7 Future Trends: Correlative Microscopy of Fluorescent Images Acquired by Confocal Laser Scanning Microscopy
References
8 Fluorescence Resonance Energy Transfer and Applications
8.1 What is FRET?
8.2 Why FRET Can Be Useful
8.3 How FRET Can Be Measured
8.4 Emerging Applications Including Novel FRET Probes
8.5 Advanced FRET Methods
References
9 Monitoring Molecular Dynamics in Live Cells Using Fluorescence Photobleaching
9.1 Introduction
9.2 Photobleaching Theory
9.3 Dynamics of Macromolecules
9.4 Photobleaching Measurements with Confocal Microscope
9.5 Photobleaching Applications
9.6 Conclusions
References
10 Time-Resolved Fluorescence in Microscopy
10.1 Introduction
10.2 Photophysics and Deactivation of Excited State
10.3 Time-Resolved Fluorescence Measurements
10.4 Time-Resolved Fluorescence in Microscopy
10.5 Conclusions
References
11 Fluorescence Correlation Spectroscopy
11.1 Introduction
11.2 Optics of Fluorescence Correlation Spectroscopy
11.3 Practical Aspects of FCS Experiments
11.4 Quantitative Evaluation of FCS Measurements to Obtain Diffusion Constants and Concentration
11.5 Conclusion
References
12 Flow Cytometry
12.1 What is Flow Cytometry?
12.2 Excitation Sources
12.3 Signal Detection and Analysis
12.4 Cell Sorting
12.5 Applications of Flow Cytometry
References
13 Fluorescence in Diagnostic Imaging
13.1 Introduction
13.2 Principles of Fluorescence Applied to Medical Diagnosis
13.3 Optical Fluorescence Imaging Techniques In Vivo
13.4 Imaging of Whole-Body Biological Systems
13.5 Future Directions
14 Fluoreseenee in Clinieal Diagnosis
14.1 Introduction
14.2 Applications of Fluorescence in Clinical Biochemistry
14.3 Fluorescence in Pathology and Cancer Diagnostics
References
15 Immunochemical Detection of Analytes by Using Fluorescence
15.1 Introduction: Definition and General Principles of Immunoassays
15.2 Immunoassay Types and Formats
15.3 Types of Analytes
15.4 Steady-State Fluorescence Immunoassays
15.5 Time-Resolved and Kinetic Approaches as Tools for Elimination of Fluorescence Background
15.6 Recent Advances in Immunoassay Signal Enhancement and Throughput
References
16 Membrane Organization
16.1 Concepts of Organization of Biological Membranes
16.2 Fluorescence Methods to Study Membrane Organization
16.3 Model Membranes
16.4 Membrane Organization in Cells
References
17 Probing Kinetics of Ion Pumps Via Voltage-Sensitive Fluorescent Dyes
17.1 Introduction: Voltage-Dependent Physiological Processes, Ion Pumps, and Channels
17.2 Voltage-Sensitive Dyes: Mechanisms, Response Times, and Their Relevance for Kinetic Studies
17.3 Steady-State Ion Pump Activity
17.4 Kinetics of Ion Pump Partial Reactions
17.5 Future Directions
References
Index