Fluorescence Fluctuation Spectroscopy (FFS), Part A - 1st Edition - ISBN: 9780123884220, 9780123884237

Fluorescence Fluctuation Spectroscopy (FFS), Part A, Volume 518

1st Edition

Serial Volume Editors: Sergey Tetin
eBook ISBN: 9780123884237
Hardcover ISBN: 9780123884220
Imprint: Academic Press
Published Date: 15th January 2013
Page Count: 336
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Table of Contents

Series page



Methods in Enzymology

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


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


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


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


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


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


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

1 Introduction

2 Background

3 Theory

4 Methods

5 Results

6 Conclusions


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


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


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


Author Index

Subject Index


This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume covers Fluorescence Fluctuation Spectroscopy
Contains chapters on such topics as Time-integrated fluorescence cumulant analysis, Pulsed Interleaved Excitation, and raster image correlation spectroscopy and number and brightness analysis.

Key Features

  • Continues the legacy of this premier serial with quality chapters authored by leaders in the field
  • Covers fluorescence fluctuation spectroscopy
  • Contains chapters on such topics as time-integrated fluorescence cumulant analysis, pulsed interleaved excitation, and raster image correlation spectroscopy and number and brightness analysis.


Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists


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© Academic Press 2013
Academic Press
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About the Serial Volume Editors

Sergey Tetin Serial Volume Editor