Orientation of the Chromophore Dipoles in the TOTO-DNA System

Juleon M. Schins, Alexandra Agronskaia, Bart G. de Grooth, Jan Greve

    Research output: Contribution to journalArticleAcademicpeer-review

    14 Citations (Scopus)

    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.
    Original languageEnglish
    Pages (from-to)230-237
    Number of pages8
    JournalCytometry
    Volume37
    Issue number3
    DOIs
    Publication statusPublished - 1999

    Fingerprint

    DNA
    Hydrodynamics
    Flow Cytometry
    Lasers
    Fluorescence
    Staining and Labeling
    1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-thiazole)-2-methylidene)quinolinium
    thiazole orange

    Keywords

    • TOTO
    • Orientation
    • Dipole
    • DNA
    • Fluorescence

    Cite this

    Schins, Juleon M. ; Agronskaia, Alexandra ; de Grooth, Bart G. ; Greve, Jan. / Orientation of the Chromophore Dipoles in the TOTO-DNA System. In: Cytometry. 1999 ; Vol. 37, No. 3. pp. 230-237.
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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.",
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    author = "Schins, {Juleon M.} and Alexandra Agronskaia and {de Grooth}, {Bart G.} and Jan Greve",
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    Orientation of the Chromophore Dipoles in the TOTO-DNA System. / Schins, Juleon M.; Agronskaia, Alexandra; de Grooth, Bart G.; Greve, Jan.

    In: Cytometry, Vol. 37, No. 3, 1999, p. 230-237.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Orientation of the Chromophore Dipoles in the TOTO-DNA System

    AU - Schins, Juleon M.

    AU - Agronskaia, Alexandra

    AU - de Grooth, Bart G.

    AU - Greve, Jan

    PY - 1999

    Y1 - 1999

    N2 - 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.

    AB - 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.

    KW - TOTO

    KW - Orientation

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    KW - Fluorescence

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