title: 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of high-order correlations in fluorescence fluctuation spectroscopy was developed in the late 1980s but since then has been replaced by alternative brightness analysis methods.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.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 S. Euclid Ave, Box 8231, St. Louis, Missouri 63110, USA.
authors.forename: Artem V
authors.initials: AV
authors.lastname: Melnykov
authors.forename: Kathleen B
authors.initials: KB
authors.lastname: Hall
mesh_terms: D000465:Algorithms; D004247:DNA / Q000737:chemistry; D004058:Diffusion; D005456:Fluorescent Dyes / Q000737:chemistry; D038761:Photobleaching; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
chemical_list: D005456:Fluorescent Dyes; D012235:Rhodamines; D004247:DNA