Control of the fluorescence lifetime in dye based nanoparticles
Control of the fluorescence lifetime in dye based nanoparticles
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DOI：
10.1039/d3sc05496a
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发表时间：
2024-02-29
期刊：
影响因子：
8.4
通讯作者：
Laursen，Bo W.
中科院分区：
文献类型：
--
作者：
Stenspil，Stine G.;Chen，Junsheng;Laursen，Bo W.
基于荧光染料的纳米颗粒(NPs)因其高亮度和稳定性而受到越来越多的关注。在荧光显微镜和分析中，高信号背景比和多通道检测是人们梦寐以求的。为此，时间分辨成像提供了背景抑制和光谱重叠信号的时间分离。虽然基于染料的纳米粒子和时间分辨成像被广泛单独使用，但两者的结合并不常见。这可能是因为基于染料的纳米粒子通常表现出较短的且非单指数寿命。纳米粒子中染料的寿命信号质量较低是由于聚集引起的猝灭(ACQ)和能量向暗态的迁移造成的。在这里，我们报告了一个通过使用小分子离子隔离晶格(SMILES)的概念来防止ACQ的解决方案。此外，FRET对染料的结合将激子锁定在FRET受体上，从而提供对荧光寿命的控制。我们展示了含有几%罗丹明和二氮杂杂三角基团受体的微笑纳米粒子是如何在菁染料的作用下产生相同的发射光谱和高的量子产率，但荧光寿命却截然不同，分别为3 ns和26 ns。在荧光寿命成像中，这两个光谱相同的纳米粒子很容易在单个粒子水平上区分开来。基于染料的纳米颗粒的掺杂方法提供了可预测的荧光寿命，并允许这些明亮的成像试剂用于时间分辨成像检测模式。
Fluorescent dye based nanoparticles (NPs) have received increased interest due to their high brightness and stability. In fluorescence microscopy and assays, high signal to background ratios and multiple channels of detection are highly coveted. To this end, time-resolved imaging offers suppression of background and temporal separation of spectrally overlapping signals. Although dye based NPs and time-resolved imaging are widely used individually, the combination of the two is uncommon. This is likely due to that dye based NPs in general display shortened and non-mono-exponential lifetimes. The lower quality of the lifetime signal from dyes in NPs is caused by aggregation caused quenching (ACQ) and energy migration to dark states in NPs. Here, we report a solution to this problem by the use of the small-molecule ionic isolation lattices (SMILES) concept to prevent ACQ. Additionally, incorporation of FRET pairs of dyes locks the exciton on the FRET acceptor providing control of the fluorescence lifetime. We demonstrate how SMILES NPs with a few percent rhodamine and diazaoxatriangulenium FRET acceptors imbedded with a cyanine donor dye give identical emission spectra and high quantum yields but very different fluorescence lifetimes of 3 ns and 26 ns, respectively. The two spectrally identical NPs are easily distinguished at the single particle level in fluorescence lifetime imaging. The doping approach for dye based NPs provides predictable fluorescence lifetimes and allows for these bright imaging reagents to be used in time-resolved imaging detection modalities.