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On the Analysis of High Order Moments of Fluorescence Fluctuations

Overview
Journal Biophys J
Publisher Cell Press
Specialty Biophysics
Date 1990 Feb 1
PMID 2317556
Citations 53
Authors
H Qian
E L Elson
Affiliations
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Abstract

A simple, straightforward analysis to characterize the distribution of aggregate sizes in a reversible aggregation system at equilibrium is presented. The method, an extension of fluorescence correlation spectroscopy (FCS), is based on measurements of higher order moments of spontaneous fluctuations of fluorescence intensity emitted from a defined open region of the sample. These fluctuations indicate fluctuations of the numbers of the fluorescent molecules in the observation region. Shot noise resulting from the random character of fluorescence emission and from the photoelectric detection system is modeled as a Poisson distribution and is subtracted from the measured photon count fluctuation moments to yield the desired fluorescence fluctuation moments. This analysis can also be used to estimate the fraction of immobile fluorophores in FCS measurements.

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