Article

Journal of Fluorescence

, Volume 15, Issue 3, pp 415-422

First online:

Statistical Analysis of Diffusion Coefficient Determination by Fluorescence Correlation Spectroscopy

  • Jörg EnderleinAffiliated withInstitute for Biological Information Processing 1 & 2 Email author 
  • , Ingo GregorAffiliated withInstitute for Biological Information Processing 1 & 2
  • , Digambara PatraAffiliated withInstitute for Biological Information Processing 1 & 2
  • , Jörg FitterAffiliated withInstitute for Biological Information Processing 1 & 2

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Abstract

Fluorescence correlation spectroscopy (FCS) has become an important and widely used technique for many applications in physics, chemistry, and biology. The parameter most frequently addressed by FCS is the diffusion of molecules in solution. Due to the highly non-linear connection between the diffusion coefficient and a measured autocorrelation function, it is extremely difficult to analyse the accuracy of the diffusion-coefficient determination in a FCS experiment. Here, we present a simplified analysis based on some general maximum-likelihood considerations, and numerical result are given for the dependence of the accuracy of the diffusion-coefficient determination on sample concentration, brightness, and measurement time. Optimal concentration values for performing FCS are found.

Key Words

Fluorescence correlation spectroscopy diffusion coefficient statistical accuracy