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Two-photon excitation is combined with dual-color cross-correlation spectroscopy. The technique is experimentally demonstrated by measuring the kinetics of DNA cleavage by restriction endonuclease EcoRI.
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Heinze K.G., Koltermann A., Schwille P. Simultaneous two-photon excitation of distinct labels for dual-color fluorescence cross-correlation analysis. Proc Natl Acad Sci USA. 97:2000;10377-10382. Two-photon excitation is combined with dual-color cross-correlation spectroscopy. The technique is experimentally demonstrated by measuring the kinetics of DNA cleavage by restriction endonuclease EcoRI.
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Two-photon excitation is combined with image cross-correlation spectroscopy. Several applications, both in vitro and on live cells, are described.
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This paper describes an extensive study of DsRed, including measurement of a shift in fluorescence emission with excitation intensity, correlation of light-induced flickering as a function of excitation intensity and pH, FCS diffusion analysis, two-color excitation to depopulate the dark state(s) and two-photon excitation.
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In one of the first applications of FCS to intracellular dynamics, the concentrations and translational motions of GFP constructs in single plant cells are characterized.
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Chirico G., Bettati S., Mozzarelli A., Chen Y., Müller J.D., Gratton E. Molecular heterogeneity of O-acetylserine sulfhydrylase by two-photon excited fluorescence fluctuation spectroscopy. Biophys J. 80:2001;1973-1985.
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An extensive characterization of the use of PCH analysis to monitor intracellular GFP is presented.
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The human estrogen receptor α dimer binds a single SRC-1 coactivator molecule with an affinity dictated by agonist structure
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The use of PCH to determine the concentrations and specific brightness values for multiple species is described in detail, with particular emphasis on defining optimal detection conditions and on the role of the statistical accuracy of the histogram.
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Müller J.D., Chen Y., Gratton E. Resolving heterogeneity on the single molecular level with the photon-counting histogram. Biophys J. 78:2000;474-486. The use of PCH to determine the concentrations and specific brightness values for multiple species is described in detail, with particular emphasis on defining optimal detection conditions and on the role of the statistical accuracy of the histogram.
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