Mechanism of ethidium bromide fluorescence enhancement on binding to nucleic acids

Olmsted, J.; Kearns, David R.

doi:10.1021/bi00635a022

Cited by 528 publications

(322 citation statements)

References 43 publications

Supporting

Mentioning

284

Contrasting

Order By: Relevance

“…Spectrally resolved FLIM Q. S. Hanley S85 dequenching process similar to EB (Olmsted & Kearns 1977;Jones et al 1978;Leupin et al 1985). These data (figure 2) were originally published as plots of phase and modulation lifetime spectra.…”

Section: Ab-plotsmentioning

confidence: 99%

Spectrally resolved fluorescent lifetime imaging

Hanley

2008

J. R. Soc. Interface.

View full text Add to dashboard Cite

Placing an imaging spectrograph or related components capable of generating a spectrum between a microscope and the image intensifier of a conventional fluorescence lifetime imaging (FLIM) system creates a spectrally resolved FLIM (SFLIM). This arrangement provides a number of opportunities not readily available to conventional systems using bandpass filters. The examples include: simultaneous viewing of multiple fluorophores; tracking of both the donor and acceptor; and observation of a range of spectroscopic changes invisible to the conventional FLIM systems. In the frequency-domain implementation of the method, variation in the fractional contributions from different fluorophores along the wavelength dimension can behave as a surrogate for a frequency sweep or spatial variations while analysing fluorophore mixtures. This paper reviews the development of the SFLIM method, provides a theoretical and practical overview of frequency-domain SFLIM including: presentation of the data; manifestations of energy transfer; observation of multiple fluorophores; and the limits of single frequency methods.

show abstract

Section: Ab-plotsmentioning

confidence: 99%

Spectrally resolved fluorescent lifetime imaging

Hanley

2008

J. R. Soc. Interface.

View full text Add to dashboard Cite

show abstract

“…an alcohol, whereas the fluorescence emission wavelength does not vary appreciably. This behaviour cannot be explained satisfactorily in terms of normal solvent-induced reordering in the excited state [20]. However, it is accounted for in the analysis of LeBret and Chalvet [23] by specific interactions between solvent molecules and the phenanthridiniuin ring.…”

Section: Poly[d(g-c)] (----) and Poly[d(a-t)] Po[y(d(a-t/] ()mentioning

confidence: 98%

“…For ethidium bromide bound to calf thymus DNA the enhancement has been reported to be around 20 [3] or 12 [20], under different experimental conditions.…”

Section: Poly[d(g-c)] (----) and Poly[d(a-t)] Po[y(d(a-t/] ()mentioning

confidence: 99%

“…The interaction of ethidium bromide with water was also studied by LeBret and Chalvet [23]. They examined the effect of this interaction on the optical absorption of the dye, in order to explain the solvent dependence reported by some authors [20,23]. It has been observed that the visible absorption band of ethidium bromide shifts towards longer wavelength when the dye is dissolved in a solvent of lower polarity than water, e.g.…”

Section: Poly[d(g-c)] (----) and Poly[d(a-t)] Po[y(d(a-t/] ()mentioning

confidence: 99%

See 1 more Smart Citation

Binding of Ethidium Bromide to Self‐Complementary Deoxydinucleotides

Doglia

Gräslund

Ehrenberg

1983

European Journal of Biochemistry

View full text Add to dashboard Cite

Optical spectra titrations were performed with ethidium bromide and the self-complementary deoxydinucleotides pdCpdC, pdGpdC, pdTpdA, and pdApdT. The titrations were performed in 7.5 mM phosphate buffer, pH 7.0, at 7 "C, and with varying dinucleotide concentrations always in large excess of the dye concentration. Well-defined isosbestic points were present in each titration after correction for dinucleotide light absorption.The binding curves were evaluated in terms of simple bimolecular or termolecular reaction models. The bimolecular reaction model gave a significantly better fit to the experimental data, judging from a computerized nonlinear least-squares fitting procedure. The following equilibrium constants were obtained:KA.-r = 350 M -From these data the absorption spectra of the completely bound dye were evaluated. These spectra showed bathchromic shifts of their maxima, increasing with the magnitude of K .Fluorescence spectra of ethidium bromide/dinucleotide mixtures were recorded under conditions similar to those for absorption spectra. From the known equilibrium constants the contributions of the bound dye could be estimated. The following fluorescence enhancements Ib/If were found: Ic-GjIf = 6.5; = 3.0; Iz-A/If = 2.0; I tFrom our results, in relation to other theoretical and experimental studies, we conclude that electrostatic phosphate-dye interactions give rise to a major part of the binding energy, which varies with dinucleotide geometry. The more strongly bound complexes exhibit less exposure of the dye to the solvent.The interaction of the cationic dye ethidium bromide with nucleic acids has been studied extensively over the past 20 years [I -61. Many of the studies have dealt with the intercalation process because of its potential biological significance. Basesequence specificity for cationic dyes in DNA has been observed for a number of acridine dyes [7-101, whereas for ethidium bromide only indirect evidence is available from binding competition experiments [I I]. In simple model systems, such as deoxydinucleotides or deoxytetranucleotides, on the other hand it has been quite clear from qualitative considerations that ethidium bromide exhibits a preference for binding to pyrimidine (3' + 5')purine sequences over purine-(3' -+ 5')pyrimidine isomers [12, 131. We present here a quantitative study of the binding of ethidium bromide to self-complementary deoxydinucleotides, which has allowed the evaluation of the equilibrium constants for the different complexes. From this knowledge it was possible to determine the spectroscopic properties of the bound dye in each complex. These results in turn lead to some further understanding of the nature of dye-dinucleotide interactions in these complexes. MATERIALS AND METHODSEthidium bromide was purchased from Sigma Chemical Company and used without further purification. A stock solution of 0.5 mM was prepared in 7.5 mM phosphate buffer, pH 7.0.Ethidium bromide concentrations were determined spectrophotometrically in the same buffer according to the molar ...

show abstract

“…Among the many different mechanisms proposed to explain this enhancement, the intercalation of ethidium bromide into DNA is the most plausible one. 27,32 More recently, the fluorescence spectra of 4-[4-(dimethylamino)styryl]-1-dodecyl pyridinium bromide (DMASDPB) mixed with DNA-CTMA were studied by Grote et al 24 As shown in Figure 6, the film containing the lowest level of dye presented the greatest fluorescence intensity. Compartmentalization of the dye molecules in the DNA-CTMA matrix must be the reason for this observation.…”

Section: Fluorescence Of Dye-doped Dnasmentioning

confidence: 99%

Optical, electro-optic and optoelectronic properties of natural and chemically modified DNAs

Kwon

Choi

Jin

2012

Polym J

View full text Add to dashboard Cite

This article reviews recent findings on the optical, electro-optic and optoelectronic properties of natural and modified DNAs. When the sodium (Na þ ) ions of DNA are replaced with long alkyl quaternary ammonium (Q þ ) ions, the resulting compositions (Q þ DNA À ) are organic-soluble, and thin films produced using these materials reveal many interesting optical and optoelectronic properties. These films tend to form well-structured supramolecular assemblies. In contrast, natural DNAs are water-soluble and hygroscopic. DNAs are strong absorbers of UV wavelengths in the region of 260 nm. The Q þ DNA À films are excellent dielectrics that can be utilized as insulating layers in organic thin film transistors. Chemical modification of the Q þ parts results in many interesting structures that can be used in a wide variety of optical and optoelectronic devices. This review specifically deals with the optical and fluorescence properties of, organic lasing composites, the nonlinear optical characteristics of, light-emitting diodes, and photovoltaic cells based on natural and modified DNAs.

show abstract

Mechanism of ethidium bromide fluorescence enhancement on binding to nucleic acids

Cited by 528 publications

References 43 publications

Spectrally resolved fluorescent lifetime imaging

Spectrally resolved fluorescent lifetime imaging

Binding of Ethidium Bromide to Self‐Complementary Deoxydinucleotides

Optical, electro-optic and optoelectronic properties of natural and chemically modified DNAs

Contact Info

Product

Resources

About