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

Revival of high-order fluorescence correlation analysis: generalized theory and biochemical applications.
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DOI:
10.1021/jp906539k
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发表时间:
2009-11-26
期刊:
The journal of physical chemistry. B
影响因子:
--
通讯作者:
Hall KB
Hall KB
中科院分区:
其他
文献类型:
--
作者:
Melnykov AV;Hall KB

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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of high order correlations in fluorescence fluctuation spectroscopy was developed in the late 1980s but since then has been replaced by alternative brightness analysis methods.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.
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