Article

Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration

  1. Jörg Enderlein Dr.1,
  2. Ingo Gregor Dr.1,
  3. Digambara Patra Dr.1,2,
  4. Thomas Dertinger1,
  5. U. Benjamin Kaupp Dr.1

Article first published online: 7 NOV 2005

DOI: 10.1002/cphc.200500414

Issue

ChemPhysChem

ChemPhysChem

Volume 6, Issue 11, pages 2324–2336, November 11, 2005

Additional Information

How to Cite

Enderlein, J., Gregor, I., Patra, D., Dertinger, T. and Kaupp, U. B. (2005), Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration. ChemPhysChem, 6: 2324–2336. doi: 10.1002/cphc.200500414

Author Information

  1. 1

    Institute for Biological Information Processing 1, Forschungszentrum Jülich, 52425 Jülich, Germany, Fax: (+49) 2461-614216

  2. 2

    Current address: Department of Physics, School of Science and Engineering, Okubo 3-4-1, Shinjuku-ku, Tokyo, 169-8555, Japan

Publication History

  1. Issue published online: 7 NOV 2005
  2. Article first published online: 7 NOV 2005
  3. Manuscript Received: 27 JUL 2005

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Keywords:

  • confocal microscopy;
  • fluorescence spectroscopy;
  • laser spectroscopy;
  • photophysics;
  • single-molecule spectroscopy

Abstract

Fluorescence correlation spectroscopy (FCS) has become an important tool for measuring diffusion, concentration, and molecular interactions of cellular components. The interpretation of FCS data critically depends on the measurement set-up. Here, we present a rigorous theory of FCS based on exact wave-optical calculations. Six of the most important optical and photophysical factors that influence FCS are studied: fluorescence anisotropy, cover-slide thickness, refractive index of the sample, laser-beam geometry, optical saturation, and pinhole adjustment. Our theoretical framework represents a general attempt to link all relevant parameters of the experimental set-up with the measured correlation function.

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