A palette of background-free tame fluorescent probes for intracellular multi-color labelling in live cells.
A palette of background-free tame fluorescent probes for intracellular multi-color labelling in live cells.
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DOI：
10.1039/c7sc04716a
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发表时间：
2018-02-28
期刊：
影响因子：
8.4
通讯作者：
Chang YT
中科院分区：
文献类型：
--
作者：
Alamudi SH;Su D;Lee KJ;Lee JY;Belmonte-Vázquez JL;Park HS;Peña-Cabrera E;Chang YT
设计了一种无背景的温和荧光探针调色板，并将其应用于活细胞的细胞内多色标记。使用小型荧光探针在其原生环境中可视化多个细胞内设备的多色标记技术仍然具有挑战性。该方法要求在非均相生物体系中进行正交和生物相容性偶联反应，荧光背景噪声最小。在这里，我们提出了一个调色板的BODIPY探针含有叠氮化物和环青的部分在活细胞中的无铜点击化学。这种探针被称为“驯服探针”，具有高度渗透性和特异性，不会在细胞中留下背景噪音。这些探针通过优化亲脂性、水溶性和荷电范德华表面积进行合理设计，使我们能够快速高效地同时标记细胞内目标成分。我们表明，这些探针不仅能够标记细胞器和工程蛋白，而且能够通过在各种细胞类型中使用代谢寡糖工程技术显示细胞内糖缀合物的动力学。因此，本研究的结果为本地系统在高时空方式下的多光谱标记策略提供了灵活性。
A palette of background-free tame fluorescent probes were designed and applied to intracellular multi-color labelling in live cells. A multi-color labelling technique for visualizing multiple intracellular apparatuses in their native environment using small fluorescent probes remains challenging. This approach requires both orthogonal and biocompatible coupling reactions in heterogeneous biological systems with minimum fluorescence background noise. Here, we present a palette of BODIPY probes containing azide and cyclooctyne moieties for copper-free click chemistry in living cells. The probes, referred to as ‘tame probes’, are highly permeable and specific in nature, leaving no background noise in cells. Such probes, which are rationally designed through optimized lipophilicity, water solubility and charged van der Waals surface area, allow us to demonstrate rapid and efficient concurrent multi-labelling of intracellular target components. We show that these probes are capable of not only labelling organelles and engineered proteins, but also showing the intracellular glycoconjugates’ dynamics, through the use of metabolic oligosaccharide engineering technology in various cell types. The results demonstrated in this study thus provide flexibility for multi-spectral labelling strategies in native systems in a high spatiotemporal manner.
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