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000058183 0247_ $$2DOI$$a10.1002/cphc.200600638
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000058183 084__ $$2WoS$$aChemistry, Physical
000058183 084__ $$2WoS$$aPhysics, Atomic, Molecular & Chemical
000058183 1001_ $$0P:(DE-Juel1)VDB15798$$aDertinger, T.$$b0$$uFZJ
000058183 245__ $$aTwo focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements
000058183 260__ $$aWeinheim$$bWiley-VCH Verl.$$c2007
000058183 300__ $$a433 - 443
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000058183 440_0 $$010582$$aChemPhysChem$$v8$$x1439-4235$$y3
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000058183 520__ $$aWe present a new method to measure absolute diffusion coefficients at nanomolar concentrations with high precision. Based on a modified fluorescence correlation spectroscopy (FCS)-setup, this method is improved by introducing an external ruler for measuring the diffusion time by generating two laterally shifted and overlapping laser foci at a fixed and known distance. Data fitting is facilitated by a new two-parameter model to describe the molecule detection function (MDF). We present a recorded MDF and show the excellent agreement with the fitting model. We measure the diffusion coefficient of the red fluorescent dye Atto655 under various conditions and compare these values with a value achieved by gradient pulsed field NMR (GPF NMR). From these measurements we conclude, that the new measurement scheme is robust against optical and photophysical artefacts which are inherent to standard FCS. With two-focus-FCS, the diffusion coefficient of 4.26 x 10(-6) cm2s(-1) for Atto655 in water at 25 degrees C compares well with the GPF NMR value of 4.28 x 10(-6) cm2s(-1).
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000058183 650_2 $$2MeSH$$aDiffusion
000058183 650_2 $$2MeSH$$aSensitivity and Specificity
000058183 650_2 $$2MeSH$$aSolutions
000058183 650_2 $$2MeSH$$aSpectrometry, Fluorescence: methods
000058183 650_2 $$2MeSH$$aViscosity
000058183 650_7 $$00$$2NLM Chemicals$$aSolutions
000058183 650_7 $$2WoSType$$aJ
000058183 65320 $$2Author$$adiffusion coefficients
000058183 65320 $$2Author$$afluorescence spectroscopy
000058183 65320 $$2Author$$afluorescent dyes
000058183 65320 $$2Author$$atime-resolved spectroscopy
000058183 7001_ $$0P:(DE-HGF)0$$aPacheco, V.$$b1
000058183 7001_ $$0P:(DE-Juel1)VDB57288$$avon der Hocht, I.$$b2$$uFZJ
000058183 7001_ $$0P:(DE-HGF)0$$aHartmann, R.$$b3
000058183 7001_ $$0P:(DE-Juel1)VDB27456$$aGregor, I.$$b4$$uFZJ
000058183 7001_ $$0P:(DE-Juel1)VDB8451$$aEnderlein, J.$$b5$$uFZJ
000058183 773__ $$0PERI:(DE-600)2025223-7$$a10.1002/cphc.200600638$$gVol. 8, p. 433 - 443$$p433 - 443$$q8<433 - 443$$tChemPhysChem$$v8$$x1439-4235$$y2007
000058183 8567_ $$uhttp://dx.doi.org/10.1002/cphc.200600638
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000058183 9141_ $$y2007
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000058183 9201_ $$0I:(DE-Juel1)VDB804$$d31.12.2008$$gINB$$kINB-1$$lZelluläre Biophysik$$x0
000058183 9201_ $$0I:(DE-Juel1)VDB805$$d31.12.2008$$gINB$$kINB-2$$lMolekulare Biophysik$$x1
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