Fluorescence Fluctuation Spectroscopy (FFS), Part A

Series page

Contributors

Preface

Methods in Enzymology

Chapter One. 40 Years of FCS: How It All Began

References

Chapter Two. Brief Introduction to Fluorescence Correlation Spectroscopy

1 Introduction

2 Conceptual Basis and Theory of FCS

3 Measurements Based on Fluctuation Amplitudes

4 Experimental Measurements

5 Close Relationship Between FCS and Fluorescence Photobleaching

6 Scanning and Imaging Approaches to FCS

7 A Brief and Partial Survey of Applications

8 Summary

References

Chapter Three. Dual-Color Fluorescence Cross-Correlation Spectroscopy with Continuous Laser Excitation in a Confocal Setup

1 Introduction

2 FCS in a Nutshell

3 The Dual-Color FCCS Extension

4 Data Analysis

5 Ligand Binding at the Cell Surface

6 Conclusion

References

Chapter Four. Brightness Analysis

1 Introduction

2 Background

3 Overview of Analysis Techniques

4 Brightness Analysis

5 Experimental Considerations

6 Brightness and Geometry

7 Summary

References

Chapter Five. Time-Integrated Fluorescence Cumulant Analysis and Its Application in Living Cells

1 Introduction

2 Theory and Implementation of TIFCA

3 Application of TIFCA

4 Conclusion

References

Chapter Six. Raster Image Correlation Spectroscopy and Number and Brightness Analysis

1 Introduction

2 A Conceptual Overview of Fluctuation Methods

3 What Is Different in the RICS Method?

4 RICS and Cross-RICS

5 PCH and Amplitude Fluctuation Analysis

6 N&B and Cross-NB

7 Simulations for Cross-RICS and Cross-N&B

8 Applications of RICS and N&B to Detect Molecular Complexes in Cells

9 Calibration Measurements using EGFP and mCherry and Effective Bleedthrough

10 Difference of Distribution in the Nucleus Versus Cytoplasm of Dynamin-2a

11 Conclusions and Future Prospects

12 Materials and Methods

References

Chapter Seven. Global Analysis in Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Microscopy

1 Introduction

2 Background

3 Theory

4 Methods

5 Results

6 Conclusions

References

Chapter Eight. Dual-Focus Fluorescence Correlation Spectroscopy

1 Introduction

2 Dual-Focus Fluorescence Correlation Spectroscopy and Translational Diffusion

3 Flow Measurements with 2fFCS

4 2fFCS Measurements in Lipid Membranes

5 Summary

References

Chapter Nine. Pulsed Interleaved Excitation

1 Introduction: The Need for PIE

2 How Does PIE Work?

3 Juggling Photons with PIE

4 Simple and Quantitative Correlation Analysis with PIE

5 Quantifying FRET with PIE–FRET–FCCS

6 Accurate Single-Pair FRET with MFD–PIE

7 Scanning FCCS with PIE

8 Glancing at the Future for Multicolor Spectroscopy

References

Chapter Ten. Image Correlation Spectroscopy: Mapping Correlations in Space, Time, and Reciprocal Space

1 Introduction

2 Theory of ICS

3 Procedures for STICS and kICS

4 Conclusions

References

Author Index

Subject Index