FLUORESCENCE ENERGY TRANSFER BETWEEN DIMETHYLDIAZAPEROPYRENIUM DICATION AND ETHIDIUM INTERCALATED IN POLY d(A‐T)

Mergny, Jean‐Louis; Slama‐Schwok, Anny; Montenay-Garestier, Thérèse; Rougée, Michel; Hélène, Claude

doi:10.1111/j.1751-1097.1991.tb03670.x

Cited by 19 publications

(9 citation statements)

References 13 publications

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“…Energy transfer between two different dye molecules bound to the same dsDNA fragment has been observed in a number of instances. Efficient Forster fluorescence energy transfer was seen when dimethyldiazaperopyrenium (donor) and ethidium (acceptor) were bound to poly d(A-T) (20). Similar observations were reported on energy transfer between dsDNAbound thiazole orange (donor) and ethidium homodimer (acceptor) (21) and Hoechst 33342 (donor) and acridine orange (acceptor) (22).…”

Section: Discussionsupporting

confidence: 67%

Heterodimeric DNA-binding dyes designed for energy transfer: synthesis and spectroscopic properties

1993

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Heterodimeric dyes are described which bind tightly to double-stranded (dsDNA) with large fluorescence enhancements. These dyes are designed to exploit energy transfer between donor and acceptor chromophores to tune the separation between excitation and emission wavelengths. The dyes described here absorb strongly at the 488 nm argon ion line, but emit at different wavelengths, and can be applied to multiplex detection of various targets. The chromophores in these dyes, a thiazole orange-thiazole blue heterodimer (TOTAB), two different thiazole orange-ethidium heterodimers (TOED1 and TOED2), and a fluorescein-ethidium heterodimer (FED), are in each case linked through polymethyleneamine linkers. The emission maxima of the DNA-bound dyes lie at 662 (TOTAB), 614 (TOED 2), and 610 nm (FED). The dyes showed a > 100 fold enhancement of the acceptor chromophore fluorescence on binding to dsDNA and no sequence selectivity. In comparison with direct 488 nm excitation of the constituent monomeric dyes, in the heterodimers the fluorescence of the acceptor chromophores was greatly enhanced and the emission of the donor chromophores quenched by over 90%. The acceptor emission per DNA-bound dye molecule was constant from 100 DNA bp:dye to 20 bp:dye and decreased sharply at higher dye:DNA ratios.

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Section: Discussionsupporting

confidence: 67%

Heterodimeric DNA-binding dyes designed for energy transfer: synthesis and spectroscopic properties

1993

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“…17 and references therein, and Ref. 18). Unlike these earlier reports of energy transfer rates lower than expected from the Forster distance, we find that the experimentally observed transfer rates for 8 different donoracceptor pairs are comparable to those calculated from spectral parameters if we use a value of 1.25 for K* and a refractive index of I .5.…”

Section: Resultscontrasting

confidence: 51%

Fluorescence energy transfer in one dimension: Frequency‐domain fluorescence study of DNA–fluorophore complexes

Maliwal¹,

Kuśba²,

Lakowicz³

1995

Biopolymers

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Fluorescence resonance energy transfer among linear DNA bound fluorophores was carried out to study the process in one dimension. The donor fluorescence intensity decays in the case of energy transfer in one dimension are stretched exponential and show exp[-(t/tau)1/6] time dependence, which results in an initial more rapid decay and subsequent slower decay at long times when compared to those in higher dimensions. DNA-bound 4',6'-diamidino-2-phenyl indole (DAPI), acridine orange (AO), and ethidium bromide (EB) were used as donors. The acceptors were in the case of DAPI AO and EB; in the case of AO nile blue (NB), methylene blue (MB), and crystal violet (CV); and NB, MB, and oxazine 750 in the case of EB. As expected, the donor intensity decays became highly heterogeneous upon energy transfer and were characterized by the simultaneous presence of both highly and marginally quenched donors. The intensity decays for all three donors in the presence of various acceptors are satisfactorily described by the Förster model of energy transfer in one dimension. The intensity decays also allow for clear rejection of a two- or three-dimensional model. The experimentally recovered critical Förster distances (R0) ranged between 37 A in the case of DAPI and EB to 70 A in the case of AO and CV donor-acceptor pairs. These recovered R0 values compare reasonably with those calculated from spectral properties if we use values of 1.25 for k2, and 1.5 for the refractive index of DNA. The k2 value will be even higher, between 1.5 and 2.0, if the consensus DNA refractive index of 1.75 is used. These k2 values strongly suggest that the dipoles of the acceptor chromophores when bound to DNA are not randomly oriented but are aligned preferentially in plane.

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“…This result is typical of multiexponential intensity decay (26–28) and indicates that the Ho decay becomes more heterogeneous or nonexponential in the presence of 7‐AAD. Energy transfer to multiple acceptors is known to result in multiexponential or nonexponential decays for donors bound to DNA (13, 29, 30).…”

Section: Resultsmentioning

confidence: 99%

Fluorescence lifetime imaging of nuclear DNA: Effect of fluorescence resonance energy transfer

et al. 2000

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Background DNA fluorescence dyes have been used to study DNA dynamics, chromatin structure, and cell cycle analysis. However, most microscopic fluorescence studies of DNA use only steady‐state measurements and do not take advantage of the additional information content of the time‐resolved fluorescence. In this paper, we combine fluorescence imaging of DNA with time‐resolved measurements to examine the proximity of donors and acceptors bound to chromatin. Methods We used frequency‐domain fluorescence lifetime imaging microscopy to study the spatial distribution of DNA‐bound donors and acceptors in fixed 3T3 nuclei. Over 50 cell nuclei were imaged in the presence of an AT‐specific donor, Hoechst 33258 (Ho), and a GC‐specific acceptor, 7‐aminoactinomycin D (7‐AAD). Results The intensity images of Ho alone showed a spatially irregular distribution due to the various concentrations of DNA or AT‐rich DNA throughout the nuclei. The lifetime imaging of the Ho‐stained nuclei was typically flat. Addition of 7‐AAD decreased the fluorescence intensity and lifetime of the Ho‐stained DNA. The spatially dependent phase and modulation values of Ho in the presence of 7‐AAD showed that the Ho decay becomes nonexponential, as is expected for a resonance energy transfer (RET) with multiple acceptors located over a range of distances. In approximately 40 nuclei, the intensity and lifetime decrease was spatially homogeneous. In approximately 10 nuclei, addition of 7‐AAD resulted in a spatially nonhomogeneous decrease in intensity and lifetime. The RET efficiency was higher in G2/M than in G0/1 phase cells. Conclusions Because RET efficiency depends on the average distance between Ho and 7‐AAD, data suggest that the heterogeneity of lifetimes and spatial variation of the RET efficiency are caused by the presence of highly condensed regions of DNA in nuclei. Cytometry 41:178–185, 2000 © 2000 Wiley‐Liss, Inc.

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FLUORESCENCE ENERGY TRANSFER BETWEEN DIMETHYLDIAZAPEROPYRENIUM DICATION AND ETHIDIUM INTERCALATED IN POLY d(A‐T)

Cited by 19 publications

References 13 publications

Heterodimeric DNA-binding dyes designed for energy transfer: synthesis and spectroscopic properties

Heterodimeric DNA-binding dyes designed for energy transfer: synthesis and spectroscopic properties

Fluorescence energy transfer in one dimension: Frequency‐domain fluorescence study of DNA–fluorophore complexes

Fluorescence lifetime imaging of nuclear DNA: Effect of fluorescence resonance energy transfer

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