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Publication Detail
Time-resolved fluorescence and FCS studies of dye-doped DNA
  • Publication Type:
    Journal article
  • Publication Sub Type:
    Conference Proceeding
  • Authors:
    Nicolaou N, Marsh RJ, Blacker T, Armoogum DA, Bain AJ
  • Publication date:
  • Journal:
    Proceedings of SPIE - The International Society for Optical Engineering
  • Volume:
  • Status:
  • Print ISSN:
Fluorescence lifetime, anisotropy and intensity dependent single molecule fluorescence correlation spectroscopy (I-FCS) are used to investigate the mechanism of fluorescence saturation in a free and nucleotide bound fluorophore (NR6104) in an antioxidising ascorbate buffer. Nucleotide attachment does not appreciably affect the fluorescence lifetime of the probe and there is a decrease in the rate of intersystem crossing relative to that of triplet state deactivation. The triplet state fraction is seen to plateau at 72% (G-attached) and 80% (free fluorophore) in agreement with these observations. Measurements of translational diffusion times show no intensity dependence for excitation intensities between 1 and 10 kW cm and photobleaching is therefore negligible. The dominant mechanism of fluorescence saturation is thus triplet state formation. I-FCS measurements for Rhodamine 6G in water were compared with those in the ascorbate buffer. In water the triplet fraction was saturated at considerably higher powers (45% at ca. 1.5 × 10 kW cm ) than in the ascorbate buffer (55%ca. 1 1kW cm ). © 2009 SPIE. 5 -2 3 -2 -2
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