Fluorescence correlation

Terry, B. R., Matthews, E. K., and Haseloff, J. (1995). Molecular characterization of recombinant green fluorescent protein by fluorescent correlation microscopy. Biochem. Biophys. Res. Commun. 217 21—27. 

Nelson and Zahniser used a moveable injector discharge flow apparatus in these studies, generating the HO2 from the H + O2 + M reaction. They detected both OH and OH by laser-induced fluorescence, correlating their ratio with... 

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. 

As mentioned in the introductory part of this section, quantum dots exhibit quite complex non-radiative relaxation dynamics. The non-radiative decay is not reproduced by a single exponential function, in contrast to triplet states of fluorescent organic molecules that exhibit monophasic exponential decay. In order to quantitatively analyze fluorescence correlation signals of quantum dots including such complex non-radiative decay, we adopted a fluorescence autocorrelation function including the decay component of a stretched exponential as represented by Eq. (8.11). 

With the aim of elucidating molecular dynamics in a small domain, we have constmcted several microspectroscopic systems, that is, (i) the confocal microscope with the excitation light source being a femtosecond NIR laser emitting a 35 fs pulse, and (ii) the fluorescence correlation spectroscopic system with optical tweezers. 

We have also developed a method of measurement for local temperature in microspace with a fluorescence correlation technique. Using this method, the temperature elevation at the optical trapping point due to absorption of the NIR trapping beam by solvent was quantitatively evaluated the temperature at the trapping point increased linearly with increase in the incident NIR light, and the temperature elevation coefficient was mainly dependent on two physical parameters of the solvent the absorption coefficient at 1064 nm and the 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. 

Ito, S., Toitani, N., Pan, L., Tamai, N. and Miyasaka, H. (2007) Fluorescence correlation spectroscopic study on water-soluble cadmium telluride nanocrystals fast blinking dynamics in the ps-ms region. J. Phys. Condens. Matter, 19, 486208. 

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. 

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