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European Biophysics Journal

, Volume 34, Issue 4, pp 323–334 | Cite as

Global analysis of fluorescence fluctuation data

  • Victor V. Skakun
  • Mark A. Hink
  • Anatoli V. Digris
  • Ruchira Engel
  • Eugene G. Novikov
  • Vladimir V. Apanasovich
  • Antonie J. W. G. Visser
Article

Abstract

Over the last decade the number of applications of fluorescence correlation spectroscopy (FCS) has grown rapidly. Here we describe the development and application of a software package, FCS Data Processor, to analyse the acquired correlation curves. The algorithms combine strong analytical power with flexibility in use. It is possible to generate initial guesses, link and constrain fit parameters to improve the accuracy and speed of analysis. A global analysis approach, which is most effective in analysing autocorrelation curves determined from fluorescence fluctuations of complex biophysical systems, can also be implemented. The software contains a library of frequently used models that can be easily extended to include user-defined models. The use of the software is illustrated by analysis of different experimental fluorescence fluctuation data sets obtained with Rhodamine Green in aqueous solution and enhanced green fluorescent protein in vitro and in vivo.

Keywords

Fluorescence correlation spectroscopy Global analysis Green fluorescent protein Translational diffusion Triplet state 

Abbreviations

EGFP

Enhanced green fluorescent protein

FCS

Fluorescence correlation spectroscopy

FCCS

Fluorescence cross-correlation spectroscopy

GFP

Green fluorescent protein

PBS

Phosphate-buffered saline

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Copyright information

© EBSA 2005

Authors and Affiliations

  • Victor V. Skakun
    • 1
  • Mark A. Hink
    • 2
  • Anatoli V. Digris
    • 1
  • Ruchira Engel
    • 2
  • Eugene G. Novikov
    • 3
  • Vladimir V. Apanasovich
    • 1
  • Antonie J. W. G. Visser
    • 2
  1. 1.Department of Systems AnalysisBelarusian State UniversityMinskBelarus
  2. 2.MicroSpectroscopy Centre, Laboratory of BiochemistryWageningen UniversityET WageningenThe Netherlands
  3. 3.Institut Curie, Section de RechercheParisFrance

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