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Revival of high-order fluorescence correlation analysis: generalized theory and biochemical applications.

ABSTRACT: Analysis of high-order correlations in fluorescence fluctuation spectroscopy was developed in the late 1980s but since then has been replaced by alternative brightness analysis methods. However, high-order correlation has important advantages in many experiments. We present a new cumulant-based formalism of high-order correlation that greatly simplifies data analysis. The new formalism is used to derive general expressions for variance of high-order correlations that show good agreement with experiment in a model system of fluorescently labeled DNA oligomers. A simulation of binary systems in which both diffusion time and brightness are varied illustrates clearly that high-order analysis has better sensitivity to brightness than fluorescence correlation spectroscopy (FCS). These results have implications for analysis of isomerization reactions and dual-beam FCS with flow. We also demonstrate that high-order correlations can detect photobleaching in the observation volume. The application of this formalism to many FCS-based experiments allows more accurate analysis in addition to describing more molecular parameters.

SUBMITTER: Melnykov AV

PROVIDER: S-EPMC2845455 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature

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Revival of high-order fluorescence correlation analysis: generalized theory and biochemical applications.

Melnykov Artem V AV Hall Kathleen B KB

The journal of physical chemistry. B 20091101 47

Analysis of high-order correlations in fluorescence fluctuation spectroscopy was developed in the late 1980s but since then has been replaced by alternative brightness analysis methods. However, high-order correlation has important advantages in many experiments. We present a new cumulant-based formalism of high-order correlation that greatly simplifies data analysis. The new formalism is used to derive general expressions for variance of high-order correlations that show good agreement with exp ...[more]

PMID: 19877707

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Dataset Information

Revival of high-order fluorescence correlation analysis: generalized theory and biochemical applications.

Publications

Revival of high-order fluorescence correlation analysis: generalized theory and biochemical applications.

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