Fluorescent correlation spectroscopy

Application of Fluorescence Correlation Spectroscopy to the Measurement of Local Temperature at a Small Area in Solution... 

Fluorescence intensity detected with a confocal microscope for the small area of diluted solution temporally fluctuates in sync with (i) motions of solute molecules going in/out of the confocal volume, (ii) intersystem crossing in the solute, and (hi) quenching by molecular interactions. The degree of fluctuation is also dependent on the number of dye molecules in the confocal area (concentration) with an increase in the concentration of the dye, the degree of fluctuation decreases. The autocorrelation function (ACF) of the time profile of the fluorescence fluctuation provides quantitative information on the dynamics of molecules. This method of measurement is well known as fluorescence correlation spectroscopy (FCS) [8, 9]. 

Application of Fluorescence Correlation Spectroscopy 145 Table 8.1 Local temperature deviation, extinction coefficient, thermal conductivity. 

Rigler, R. and Elson, E. S. (eds) (2001) Fluorescence Correlation Spectroscopy, Springer Series in Chemical Physics, 65, Springer, Berlin. 

Krichevsky, O. and Bonnet, G. (2002) Fluorescence correlation spectroscopy the technique and its applications. Rep. Prog. Phys., 65, 251-297. 

Elson, E. L. and Magde, D. (1974) Fluorescence correlation spectroscopy. 1. Conceptual basis and theory. Biopolymers, 13, 1-27 Elson, E. L. and Webb, W. W. (1974) Fluorescence correlation spectroscopy. 11. An experimental realization. Biopolymers, 13, 29-61. 

Masuda, A., Ushdia, K and Okamoto, T. (2005) New fluorescence correlation spectroscopy enabbng direct observation of spatiotemporal dependence of diffusion constants as an evidence of anomalous transport in extracellular matrices. Biophys.J., 88, 3584—3591. 

Doose, S., Tsay, J. M., Pinaud, F. and Weiss, S. (2005) Comparison of photophysical and colloidal properties of biocompatible semiconductor nanocrystals using fluorescence correlation spectroscopy. Anal. Chem., 77, 2235-2242. 

Application of fluorescence correlation spectroscopy to the measurement of local temperature in solutions under optical trapping condition./. Phys. Chem. B, 111, 2365-2371. 

Gregor, 1., Patra, D. and Enderlein, J. (2005) Optical saturation in fluorescence correlation spectroscopy under continuous-wave and pulsed excitation. 

Hosokawa, C., Yoshikawa, H. and Masuhara, H. (2004) Optical assembling dynamics of individual polymer nanospheres investigated by singleparticle fluorescence detection. Phys. Rev. E, 70, 061410-1-061410-7 (2005) Cluster formation of nanoparticles in an optical trap studied by fluorescence correlation spectroscopy. Phys. Rev. E, 72, 021408-1-021408-7. 

Schwille, P., Korkach, J. and Webb, W. W. (1999) Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes. Cytometry, 36, 176-182. 

Burns, A. R., Frankel, D. J. and Buranda, T. (2005) Local mobility in hpid domains of supported bilayers characterized by atomic force microscopy and fluorescence correlation spectroscopy. Biophys. J., 89, 1081-1093. 

Fluorescence Correlation Spectroscopy on Molecular Diffusion Inside and Outside a Single Living Cell 645... 

WHAT FLUORESCENCE CORRELATION SPECTROSCOPY CAN TELL US ABOUT UNFOLDED PROTEINS... 

Fluorescence correlation spectroscopy (FCS) measures rates of diffusion, chemical reaction, and other dynamic processes of fluorescent molecules. These rates are deduced from measurements of fluorescence fluctuations that arise as molecules with specific fluorescence properties enter or leave an open sample volume by diffusion, by undergoing a chemical reaction, or by other transport or reaction processes. Studies of unfolded proteins benefit from the fact that FCS can provide information about rates of protein conformational change both by a direct readout from conformation-dependent fluorescence changes and by changes in diffusion coefficient. 

IV. Advantages and Disadvantages of Using Fluorescence Correlation Spectroscopy to Study Protein Conformational Changes... 

The material presented in this chapter demonstrates the utility of fluorescence correlation spectroscopy in the study of unfolded proteins. 

Czemey P, Lehmann F, Wenzel M, Buschmann V, Dietrich A, Mohr GJ (2001) Tailor-made dyes for fluorescence correlation spectroscopy (FCS). Biol Chem 382 495-498... 

Haupts U, Maiti S, Schwille P, Webb WW (1998) Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy. Proc Natl Acad Sci USA 95 13573-13578... 

Schwille P, Kummer S, Heikal AA, Moemer WE, Webb WW (2000) Fluorescence correlation spectroscopy reveals fast optical excitation-driven intramolecular dynamics of yellow fluorescent proteins. Proc Natl Acad Sci USA 97 151-156... 

Elson, E. and Magde, D. (1974). Fluorescence correlation spectroscopy I Conceptual basis and theory. Biopolymers 13, 1-28. 

Schwille, P., Bieschke, J. and Oehlenschlager, F. (1997). Kinetic investigations by fluorescence correlation spectroscopy The analytical and diagnostic potential of diffusion studies. Biophys. Chem. 66, 211-28. 

Webb, W. (2001). Fluorescence correlation spectroscopy Inception, biophysical experimentations, and prospectus. Appl. Opt. 40, 3969-83. 

Bjemeld E.J., Foldes-Papp Z., Kail M., Rigler R., Single-molecule surface-enhanced Raman and fluorescence correlation spectroscopy of horseradish peroxidase, J. Phys. Chem. B 2002 106 1213-1218. 

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