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Structure-Fluorescence Contrast Relationship in Cyanine DNA Intercalators: Toward Rational Dye Design

机译:花青DNA嵌入剂中的结构荧光对比关系:走向合理的染料设计。

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摘要

The fluorescence enhancement mechanisms of a series of DNA stains of the oxazole yellow (YO) family have been investigated in detail using steady-state and ultrafast time-resolved fluorescence spectroscopy. The strong increase in the fluorescence quantum yield of these dyes upon DNA binding is shown to originate from the inhibition of two distinct processes: 1) isomerisation through large-amplitude motion that non-radiatively deactivates the excited state within a few picoseconds and 2) formation of weakly emitting H-dimers. As the H-dimers are not totally non-fluorescent, their formation is less efficient than isomerisation as a fluorescent contrast mechanism. The propensity of the dyes to form H-dimers and thus to reduce their fluorescence contrast upon DNA binding is shown to depend on several of their structural parameters, such as their monomeric (YO) or homodimeric (YOYO) nature, their substitution and their electric charge. Moreover, these parameters also have a substantial influence on the affinity of the dyes for DNA and on the ensuing sensitivity for DNA detection. The results give new insight into the development and optimisation of fluorescent DNA probes with the highest contrast.
机译:使用稳态和超快时间分辨荧光光谱技术详细研究了恶唑黄(YO)家族的一系列DNA染料的荧光增强机制。这些染料在DNA结合后的荧光量子产率的强劲增加表明,这是由于抑制了两个不同的过程:1)通过大幅度运动的异构化,该运动在几皮秒内非辐射地使激发态失活; 2)形成发射微弱的H-二聚体。由于H-二聚体并非完全非荧光,因此其形成比作为荧光对比机理的异构化效率低。染料形成H-二聚体并因此降低其在DNA结合时的荧光反差的倾向显示取决于它们的几个结构参数,例如它们的单体(YO)或同二聚体(YOYO)性质,它们的取代和它们的电收费。而且,这些参数还对染料对DNA的亲和力以及对随后的DNA检测灵敏度具有重要影响。结果为具有最高对比度的荧光DNA探针的开发和优化提供了新的见识。

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