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Chapter D11 - Fluorescence correlation spectroscopy

from Part D - Hydrodynamics

Published online by Cambridge University Press:  05 November 2012

Igor N. Serdyuk ,
Nathan R. Zaccai  and
Joseph Zaccai
Igor N. Serdyuk
Affiliation:
Institute of Protein Research, Moscow
Nathan R. Zaccai
Affiliation:
University of Bristol
Joseph Zaccai
Affiliation:
Institut de Biologie Structurale, Grenoble
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Summary

Historical review

1916

M. von Smoluchowski gave the first theoretical description of the amplitude and the temporal decay of number fluctuations in a diffusion system.

1972–1974

D. Magde, E. L. Elson and W. W. Webb published a rigorous formalism of fluorescence correlation spectroscopy (FCS) with its various modes of possible application, highlighting the large potential of this variant of the relaxation method.

1990

R. Rigler and coworkers made the final breakthrough for the FCS method. They reached the single-molecule detection limit by combining FCS with a confocal set-up, thus increasing the signal-to-noise ratio dramatically. By tightly focusing a laser beam and inserting a pinhole into the image plane, maximum lateral and axial resolution were achieved.

1994

M. Eigen and R. Rigler triggered an important further development by proposing the application of dual-colour cross-correlation for diagnostic purposes. The underlying idea was to separate single-labelled reaction educts from dual-labelled reaction products to discriminate against an excess of free single-labelled species and thus enhance the specificity of detection. In 1997 P. Schwille and coworkers successfully monitored a hybridisation reaction of two differently labelled oligonucleotides by the dual-colour cross-correlation technique. In 2000 K. G. Heinze and coworkers, reported the application of dual-colour two-photon cross-correlation to determine enzymatic cleavage of a DNA substrate by endonuclease.

2000 to now

FCS has evolved into a whole family of related methods sharing the basic principle of fluorescence fluctuation analysis.

Type
Chapter
Information
Methods in Molecular Biophysics
Structure, Dynamics, Function
 , pp. 505 - 516
Publisher: Cambridge University Press
Print publication year: 2007

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References

Bastianes, P. I. H., and Pepperkok, R. (2000). Observing proteins in their natural habitat: the living cell. TIBS, 25, 631–637.Google Scholar
Elson, E. L., and Magde, D. (1974). Fluorescence correlation spectroscopy. I. Conceptual basis and theory. Biopolymers, 13, 1–27.CrossRefGoogle Scholar
Haustein, E., and Schwille, P. (2003). Ultrasensitive investigations of biological systems by fluorescence correlation spectroscopy. Methods, 29, 153–166.CrossRefGoogle ScholarPubMed
Magde, D., Elson, E. L., and Webb, W. W. (1972). Thermodynamic fluctuations in a reacting system. Measurement by fluorescence correlation spectroscopy. Phys. Rev. Lett., 29, 705–708.CrossRefGoogle Scholar
Eigen, M., and Rigler, R. (1994). Sorting single molecules: application to diagnostics and evolutionary biothechnology. Proc. Natl. Acad Sci. USA, 91, 5740–5747.CrossRefGoogle Scholar
Bacia, K., and Schwille, P. (2003). A dynamic view of cellular processes by in vivo fluorescence auto- and cross-correlation spectroscopy. Methods, 29 74–85.CrossRefGoogle ScholarPubMed
Schwille, P., and Kettling, U. (2001). Analyzing single protein molecules using optical methods. Curr. Opin. Biotech., 12, 382–386.CrossRefGoogle ScholarPubMed
Schwille, P., Meyer-Almes, F. J., and Rigler, R. (1997). Dual-colour fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution. Biophys. J., 72, 1878–1886.CrossRefGoogle Scholar
Koltermann, A., Kettling, U., Stephan, J., Winkler, T., and Eigen, M. (2001). Dual-color confocal flourescence spectroscopy and its application in biotechnology. In Fluorescence Correlation Spectroscopy – Theory and Application, eds. Rigler, R. and Elson, E.. Heidelberg: Springer-Verlag.Google Scholar
Stephan, J., Dorre, K., et al. (2001). Towards a general procedure for sequencing single DNA molecules. J. Biotechnol., 86, 255–267.CrossRefGoogle ScholarPubMed
Bastianes, P. I. H., and Pepperkok, R. (2000). Observing proteins in their natural habitat: the living cell. TIBS, 25, 631–637.Google Scholar
Elson, E. L., and Magde, D. (1974). Fluorescence correlation spectroscopy. I. Conceptual basis and theory. Biopolymers, 13, 1–27.CrossRefGoogle Scholar
Haustein, E., and Schwille, P. (2003). Ultrasensitive investigations of biological systems by fluorescence correlation spectroscopy. Methods, 29, 153–166.CrossRefGoogle ScholarPubMed
Magde, D., Elson, E. L., and Webb, W. W. (1972). Thermodynamic fluctuations in a reacting system. Measurement by fluorescence correlation spectroscopy. Phys. Rev. Lett., 29, 705–708.CrossRefGoogle Scholar
Eigen, M., and Rigler, R. (1994). Sorting single molecules: application to diagnostics and evolutionary biothechnology. Proc. Natl. Acad Sci. USA, 91, 5740–5747.CrossRefGoogle Scholar
Bacia, K., and Schwille, P. (2003). A dynamic view of cellular processes by in vivo fluorescence auto- and cross-correlation spectroscopy. Methods, 29 74–85.CrossRefGoogle ScholarPubMed
Schwille, P., and Kettling, U. (2001). Analyzing single protein molecules using optical methods. Curr. Opin. Biotech., 12, 382–386.CrossRefGoogle ScholarPubMed
Schwille, P., Meyer-Almes, F. J., and Rigler, R. (1997). Dual-colour fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution. Biophys. J., 72, 1878–1886.CrossRefGoogle Scholar
Koltermann, A., Kettling, U., Stephan, J., Winkler, T., and Eigen, M. (2001). Dual-color confocal flourescence spectroscopy and its application in biotechnology. In Fluorescence Correlation Spectroscopy – Theory and Application, eds. Rigler, R. and Elson, E.. Heidelberg: Springer-Verlag.Google Scholar
Stephan, J., Dorre, K., et al. (2001). Towards a general procedure for sequencing single DNA molecules. J. Biotechnol., 86, 255–267.CrossRefGoogle ScholarPubMed

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