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. 1990 Feb;57(2):375–380. doi: 10.1016/S0006-3495(90)82539-X

On the analysis of high order moments of fluorescence fluctuations.

H Qian 1, E L Elson 1
PMCID: PMC1280678  PMID: 2317556

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Elson E. L., Webb W. W. Concentration correlation spectroscopy: a new biophysical probe based on occupation number fluctuations. Annu Rev Biophys Bioeng. 1975;4(00):311–334. doi: 10.1146/annurev.bb.04.060175.001523. [DOI] [PubMed] [Google Scholar]
  2. Frieden C. Actin and tubulin polymerization: the use of kinetic methods to determine mechanism. Annu Rev Biophys Biophys Chem. 1985;14:189–210. doi: 10.1146/annurev.bb.14.060185.001201. [DOI] [PubMed] [Google Scholar]
  3. Icenogle R. D., Elson E. L. Fluorescence correlation spectroscopy and photobleaching recovery of multiple binding reactions. I. Theory and FCS measurements. Biopolymers. 1983 Aug;22(8):1919–1948. doi: 10.1002/bip.360220808. [DOI] [PubMed] [Google Scholar]
  4. Koppel D. E., Axelrod D., Schlessinger J., Elson E. L., Webb W. W. Dynamics of fluorescence marker concentration as a probe of mobility. Biophys J. 1976 Nov;16(11):1315–1329. doi: 10.1016/S0006-3495(76)85776-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Magde D., Elson E. L., Webb W. W. Fluorescence correlation spectroscopy. II. An experimental realization. Biopolymers. 1974 Jan;13(1):29–61. doi: 10.1002/bip.1974.360130103. [DOI] [PubMed] [Google Scholar]
  6. Palmer A. G., 3rd, Thompson N. L. Molecular aggregation characterized by high order autocorrelation in fluorescence correlation spectroscopy. Biophys J. 1987 Aug;52(2):257–270. doi: 10.1016/S0006-3495(87)83213-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Petersen N. O. Scanning fluorescence correlation spectroscopy. I. Theory and simulation of aggregation measurements. Biophys J. 1986 Apr;49(4):809–815. doi: 10.1016/S0006-3495(86)83709-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Schlessinger J., Koppel D. E., Axelrod D., Jacobson K., Webb W. W., Elson E. L. Lateral transport on cell membranes: mobility of concanavalin A receptors on myoblasts. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2409–2413. doi: 10.1073/pnas.73.7.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Weissman M., Schindler H., Feher G. Determination of molecular weights by fluctuation spectroscopy: application to DNA. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2776–2780. doi: 10.1073/pnas.73.8.2776. [DOI] [PMC free article] [PubMed] [Google Scholar]

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