Fast interactions between Rh6G and dGTP in water studied by fluorescence correlation spectroscopy

Widengren, Jerker; Dapprich, Johannes; Rigler, Rudolf

doi:10.1016/s0301-0104(97)00014-1

Cited by 102 publications

(108 citation statements)

References 21 publications

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“…22,23 These states in turn reflect a range of environmental properties, including oxygen and other quencher molecule concentrations, viscosity, local redox environments, and temperature. Transient state parameters, monitored by FCS, can also be used as a measure of the extent to which Fluorescence (or Förster)…”

Section: Introductionmentioning

confidence: 99%

Transient State Monitoring by Total Internal Reflection Fluorescence Microscopy

et al. 2010

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Triplet, photo-oxidized and other photo-induced, long-lived states of fluorophores are sensitive to the local environment and thus attractive for microenvironmental imaging purposes. In this work, we introduce an approach where these states are monitored in a total-internal reflection (TIR) fluorescence microscope, via the characteristic variations of the time-averaged fluorescence occurring in response to different excitation modulation schemes. The surface-confined TIR excitation field generates a signal from the fluorescent molecules close to the glass surface. Thereby a high selectivity and low background noise is obtained, and in combination with low duty cycles of excitation, the overall photo-degradation of the fluorescent molecules of the sample can be kept low.To verify the approach, the kinetics of the triplet and radical states of the dye Rhodamine110 were imaged and analyzed in aqueous solutions at different concentrations of dissolved oxygen and of the reducing agent ascorbic acid. The experimental results were compared to data from corresponding Fluorescence Correlation Spectroscopy (FCS) measurements and simulations based on finite element analysis. The approach was found to accurately determine relative populations and dynamics of triplet and photo-oxidized states, overcoming passage time limitations seen in FCS measurements.The method circumvents the need of time resolution in the fluorescence detection, allowing simultaneous readout over the whole surface area subject to excitation. It can be applied over a broad range of concentrations and does not require a strong fluorescence brightness of the sample molecules. Given the sensitivity of the triplet and photo-oxidized states to oxygen concentrations and not the least to local redox environments we expect the approach to become an attractive tool for imaging cell metabolism.3

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Section: Introductionmentioning

confidence: 99%

Transient State Monitoring by Total Internal Reflection Fluorescence Microscopy

et al. 2010

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“…4,6,8,9,12 The dependency of fluorescence quenching on the distance between a dye and a guanine has been studied intensively by using fluorescent dyes, acridine, 9 stilbene, 13,14 pyrene, 15 and oxazine dyes. 16 Photoinduced electron transfer occurring in the fluorescence-quenching process has been ascertained by detailed investigations of the photophysical and photochemical behavior of free dyes of coumarin derivatives, 12 rhodamine derivatives, [17][18][19] [25][26][27][28] in the presence of a guanine. In these cases, the quenchable dyes acted as electron acceptors in their excited state and a guanine base acted as an electron donor.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence-Quenching Phenomenon by Photoinduced Electron Transfer between a Fluorescent Dye and a Nucleotide Base

Takasu¹,

Kurata

Yamada³

et al. 2001

ANAL. SCI.

330

267

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“…FCS is a statistical and analytical tool based on the temporal ACF of the measured fluorescence intensity F(t) emitted by a probe dye molecule (fluorophore). Fluctuations in the fluorescence intensity are characterized by the normalized temporal ACF G(τ) at time lag τ, which is given by 4,24,29,31 ( ) ( Figure 1. Schematic of our single molecule spectrometer.…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

Chowdhury¹,

Manho²

2011

Bulletin of the Korean Chemical Society

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Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

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Fast interactions between Rh6G and dGTP in water studied by fluorescence correlation spectroscopy

Cited by 102 publications

References 21 publications

Transient State Monitoring by Total Internal Reflection Fluorescence Microscopy

Transient State Monitoring by Total Internal Reflection Fluorescence Microscopy

Fluorescence-Quenching Phenomenon by Photoinduced Electron Transfer between a Fluorescent Dye and a Nucleotide Base

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

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