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- W2912302711 abstract "Cyanine dyes are amongst the most commonly used fluorescent probes in the biophysical research. The photophysics of these dyes and their interaction with DNA vary significantly based on the hydrophobic properties, structure etc. This study investigates fluorescent properties of a relatively new class of asymmetrical red cyanine dyes from Dyomics (Dy630, Dy632), which have similar spectral properties as Cy5, a widely used red cyanine dye. A comparative study of quantum yield and lifetime was done between Dy630 and Cy5 to see the potential of this dye as a protein-induced fluorescent enhancement (PIFE) probe. The effect of viscosity on cis-trans isomerization was more pronounced for Dy630 than Cy5, displaying potential to be a better red PIFE probe. Concentration-dependent fluorescence enhancement studies were done showing that these dyes interact with nucleobases (dNMPs) more strongly than Cy5. To investigate these interactions further, dye covalently attached to the DNA was observed using single-molecule fluorescence. Two fluorescence intensity states for Dy630 and Dy632 were observed, whereas one single state was observed for Cy5. This could be explained by a stronger pi- pi interaction possible due to the presence of additional aromatic ring in Dy dyes, giving rise to two modes of dye-DNA interaction. The single-molecule analysis gives a short-lived (0.87 s), strongly interacting component and a long-lived (3.6 s), weakly interacting component for Dy630. The comparison of dwell time between the sulfonated (Dy632) and non-sulfonated (Dy630) dye shows that the non-sulfonated dye is more “sticky” on DNA, which is supported by the lifetime measurements. Therefore, a thorough investigation of such interactions between biomolecule and probe is needed before performing single-molecule experiments like PIFE and Foster resonance energy transfer (FRET) to stay away from artifacts." @default.
- W2912302711 created "2019-02-21" @default.
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- W2912302711 date "2019-02-01" @default.
- W2912302711 modified "2023-09-30" @default.
- W2912302711 title "Environment-Dependent Photophysics of an Asymmetrical Cyanine" @default.
- W2912302711 doi "https://doi.org/10.1016/j.bpj.2018.11.3034" @default.
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