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翻訳と辞書　辞書検索 [ 開発暫定版 ] スポンサード リンク Fluorescence cross-correlation spectroscopy ： ウィキペディア英語版 Fluorescence cross-correlation spectroscopy Fluorescence cross-correlation spectroscopy (FCCS) was introduced by Eigen and Rigler in 1994 〔Eigen, M. and Rigler, R. Sorting single molecules: Application to diagnostics and evolutionary biotechnology. (1994) ''Proc. Natl. Acad. Sci. U.S.A.'' 91, 5740-5747.〕 and experimentally realized by Schwille in 1997.〔Schwille, P.; Myer-Almes, F.J.; Rigler, R. Dual-color fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution. (1997) ''Biophys. J.'' 72, 1878-1886.〕 It extends the fluorescence correlation spectroscopy (FCS) procedure in that it looks at the correlation between different colors rather than just the same color. In other words, coincident green and red intensity fluctuations correlate if green and red labeled particles are moving together. As a result, FCCS provides a highly sensitive measurement of molecular interactions independent of diffusion rate. This is an important advancement, given that diffusion rate depends only weakly on the size of the molecular complex. FCCS utilizes two species which are independently labeled with two differently colored fluorescent probes. These fluorescent probes are excited and detected by two different laser light sources and detectors usually labeled as “green” and “red”. Typically a confocal microscope is used to provide overlapping green and red focal volumes for excitation. The normalized cross-correlation function is defined for two fluorescent species $\backslash \; G$ and $\backslash \; R$ which are independent green, G and red, R channels as follows: $\backslash \; G\_(\backslash tau)=1+\backslash frac=\backslash frac$ where differential fluorescent signals $\backslash \; \backslash delta\; I\_G$ at a specific time, $\backslash \; t$ and $\backslash \; \backslash delta\; I\_R$ at a delay time, $\backslash \; \backslash tau$ later is correlated with each other. In the absence of spectral bleed-through, the cross-correlation function is zero for non-interacting particles. In contrast to FCS, the cross-correlation function increases with increasing numbers of interacting particles. FCCS is primarily utilized for measurements of bio-molecular interactions both in living cells and in vitro.〔Bacia, K.; Kim, S.A.; Schwille, P. Fluorescence cross-correlation spectroscopy in living cells. (2006) ''Nat. Meth.'' 3, 83-89 .〕〔Slaughter, B. D.; Unruh, J. R.; Li, R. Fluorescence fluctuation spectroscopy and imaging methods for examination of dynamic protein interactions in yeast. ''In Methods in Molecular Biology: Yeast Systems Biology.'' J.I. Castrillo and S.G. Oliver, Eds. (Springer, New York, 2011). Vol. 759, pp. 283-306.〕 It can be utilized to measure simple molecular stoichiometry’s and binding constants.〔Chen, Y. and Mueller, J.D. Determining the stoichiometry of protein heterocomplexes in living cells with fluorescence fluctuation spectroscopy. (2006) ''Proc. Natl. Acad. Sci. U.S.A.'' 104, 3147-3152.〕 It is one of the few techniques that can provide information about protein-protein interactions at a specific time and location within a living cell. In contrast to fluorescence resonance energy transfer, it does not have a distance limit for interactions. As a result, it can be utilized to probe large complexes. Nevertheless, it does require that complexes be actively diffusing through the microscope focus on a relatively short time scale (typically seconds). ==Modeling== Cross-correlation curves are modeled according to a slightly more complicated mathematical function than applied in FCS. First of all, the effective superimposed observation volume in which the G and R channels form a single observation volume, $\backslash \; V\_$ in the solution: $\backslash \; V\_=\backslash pi^(\backslash omega\_^2+\backslash omega\_^2)(\backslash omega\_^2+\backslash omega\_^2)^/2^$ where $\backslash \; \backslash omega\_^2$ and$\backslash \; \backslash omega\_^2$ are radial parameters and $\backslash \; \backslash omega\_$ and$\backslash \; \backslash omega\_$ are the axial parameters for the G and R channels respectively. The diffusion time, $\backslash \; \backslash tau\_$ for a doubly (G and R) fluorescent species is therefore described as follows: $\backslash \; \backslash tau\_=\backslash frac^2\}$ is the diffusion coefficient of the doubly fluorescent particle. The cross-correlation curve generated from diffusing doubly labelled fluorescent particles can be modelled in separate channels as follows: $\backslash \; G\_G(\backslash tau)=1+\backslash frac>)^2\}$ $\backslash \; G\_R(\backslash tau)=1+\backslash frac>)^2\}$ In the ideal case, the cross-correlation function is proportional to the concentration of the doubly labeled fluorescent complex: $\backslash \; G\_(\backslash tau)=1+\backslash frac(\backslash tau)\}>)(+)\}$ with $\backslash \; Diff\_k(\backslash tau)=\backslash frac\})(1+a^(\backslash frac\}$ The cross-correlation amplitude is directly proportional to the concentration of double-labeled (red and green) species 〔http://www.rci.rutgers.edu/~moghe/biophy%20j%2083_1184.pdf〕 抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』 ■ウィキペディアで「Fluorescence cross-correlation spectroscopy」の詳細全文を読む スポンサード リンク 翻訳と辞書 : 翻訳のためのインターネットリソース Copyright(C) kotoba.ne.jp 1997-2016. All Rights Reserved.