Rational Design of Bright Long Fluorescence Lifetime Dyad Fluorophores for Single Molecule Imaging and Detection,Journal of the American Chemical Society

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Rational Design of Bright Long Fluorescence Lifetime Dyad Fluorophores for Single Molecule Imaging and Detection
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2021-01-11 , DOI: 10.1021/jacs.0c10457
Laura Kacenauskaite ₁ , Niels Bisballe ₁ , Rebecca Mucci ₁ , Marco Santella ₁ , Tönu Pullerits ₂ , Junsheng Chen _{1,

2} , Tom Vosch ₁ , Bo W. Laursen ₁

Affiliation

Increasing demand for detecting single molecules in challenging environments has raised the bar for the fluorophores used. To achieve better resolution and/or contrast in fluorescence microscopy, it is now essential to use bright and stable dyes with tailored photophysical properties. While long fluorescence lifetime fluorophores offer many advantages in time-resolved imaging, their inherently lower molar absorption coefficient has limited applications in single molecule imaging. Here we propose a generic approach to prepare bright, long fluorescence lifetime dyad fluorophores comprising an absorbing antenna chromophore with high absorption coefficient linked to an acceptor emitter with a long fluorescence lifetime. We introduce a dyad consisting of a perylene antenna and a triangulenium emitter with 100% energy transfer from donor to acceptor. The dyad retained the long fluorescence lifetime (∼17 ns) and high quantum yield (75%) of the triangulenium emitter, while the perylene antenna increased the molar absorption coefficient (up to 5 times) in comparison to the free triangulenium dye. These triangulenium based dyads with significantly improved brightness can now be detected at the single molecule level and easily discriminated from bright autofluorescence by time-gated and other lifetime-based detection schemes.

中文翻译：

用于单分子成像和检测的明亮长荧光寿命二元组荧光团的合理设计

在具有挑战性的环境中检测单个分子的需求不断增加，这提高了所用荧光团的标准。为了在荧光显微镜中获得更好的分辨率和/或对比度，现在必须使用具有定制光物理特性的明亮且稳定的染料。虽然长荧光寿命荧光团在时间分辨成像中具有许多优势，但它们固有的较低摩尔吸收系数在单分子成像中的应用受到限制。在这里，我们提出了一种通用方法来制备明亮、长荧光寿命的二元荧光团，该荧光团包括具有高吸收系数的吸收天线发色团，该发色团与具有长荧光寿命的受体发射体相连。我们引入了由苝天线和三角发射体组成的二元组，能量从供体到受体的 100% 转移。二元组保留了三角发射体的长荧光寿命（~17 ns）和高量子产率（75%），而苝天线与游离三角染料相比，增加了摩尔吸收系数（高达 5 倍）。现在可以在单分子水平上检测到这些亮度显着提高的基于三角形的二元组，并且可以通过时间门控和其他基于寿命的检测方案轻松将其与明亮的自发荧光区分开来。

更新日期：2021-01-11

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