Orientation of the chromophore dipoles in the TOTO-DNA system

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Schins, Juleon M. and Agronskaia, Alexandra and Grooth de, Bart G. and Greve, Jan (1999) Orientation of the chromophore dipoles in the TOTO-DNA system. Cytometry Part A, 37 (3). pp. 230-237. ISSN 0196-4763

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Abstract:Background:
Flow cytometry has been applied successfully to the sizing of medium to large-sized DNA molecules, thanks to the excellent staining properties of cyanine chromophores such as TOTO (homodimer of thiazole orange) (Petty et al.: Anal Chem 67:1755-1761, 1995). The hydrodynamic flow, used to focus the sample molecules in a small laser-illuminated volume, is also responsible for their alignment, thereby allowing the determination of the TOTO-dipole orientation with respect to the DNA axis (Agronskaia et al.: Appl Opt 38:714-719, 1999). -
Methods:
We present model calculations of the fluorescence yield of TOTO-stained DNA measured in a flow-cytometric setup with high numerical aperture. The models consider different orientations of the chromophore dipoles. -
Results:
Comparison of measurement and calculation suggests that the absorption dipoles of the TOTO molecule make a mean angle of 61° with the helix axis of the DNA molecule. This mean angle can be the consequence of two binding modes. - Conclusions:
Our results indicate that any model with a significant contribution of perpendicularly-oriented chromophores fails to reproduce the experimental results.
Item Type:Article
Copyright:© 1999 Wiley-Liss
Research Group:
Link to this item:http://purl.utwente.nl/publications/60734
Official URL:http://dx.doi.org/10.1002/(SICI)1097-0320(19991101)37:3<230::AID-CYTO10>3.0.CO;2-#
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