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一种用于溶酶体和自噬溶酶体特异性标记的环定位荧光探针。

An azacyclo-localizing fluorescent probe for the specific labeling of lysosome and autolysosome.

机构信息

Institute of Intelligent Machines, Chinese Academy of Sciences, Hefei, 230031, China; State Key Laboratory of Transducer Technology, Chinese Academy of Sciences, Hefei, Anhui, 230031, China.

Institute of Intelligent Machines, Chinese Academy of Sciences, Hefei, 230031, China; School of Chemistry and Chemical Engineering, Anhui University, Hefei, 230601, China; State Key Laboratory of Transducer Technology, Chinese Academy of Sciences, Hefei, Anhui, 230031, China.

出版信息

Talanta. 2020 Aug 15;216:120941. doi: 10.1016/j.talanta.2020.120941. Epub 2020 Mar 23.

Abstract

Understanding lysosome-related physiology needs specific lysosome probes to track the biological processes of lysosome in living cells. Here, we report an azacyclo-modified fluorescent probe that has a large Stokes shift, good photostability and negligible cytotoxicity for highly specific labeling of lysosome and autolysosome in living cells. The probes with different kinds of azacyclo groups on parent dye dansyl are screened to show that dansyl-cycleanine (DNS-C) with four nitrogen atoms possesses the best lysosome-localized ability. And DNS-C as a universal tracker exhibits excellent ability for lysosome labeling in different cell lines with high overlap coefficients (≥0.90). Different from a commercially available LysoTracker, the Stokes shift of DNS-C up to 240 nm (λ = 330/570 nm), is much larger than that of LysoTracker ~20 nm (λ = 573/595 nm). More importantly, the fluorescence of DNS-C keeps still high brightness after a time-lapsed imaging for 40 min in living cells, implying its remarkable photostability for long-term tracking. In addition, DNS-C can also clearly image the autolysosome, a critical subcellular compartment, forming by the fusion of lysosome with autophagosome in autophagy. These results suggest the promising utility of our probe as a powerful tool to real-time trace physiological processes of lysosomes.

摘要

了解溶酶体相关的生理学需要特定的溶酶体探针来跟踪活细胞中溶酶体的生物学过程。在这里,我们报告了一种氮杂环修饰的荧光探针,它具有较大的斯托克斯位移、良好的光稳定性和可忽略的细胞毒性,可高度特异性地标记活细胞中的溶酶体和自溶酶体。我们筛选了在母体染料丹磺酰上具有不同氮杂环基团的探针,结果表明具有四个氮原子的丹磺酰环辛烷(DNS-C)具有最佳的溶酶体定位能力。并且作为一种通用示踪剂,DNS-C 在具有高重叠系数(≥0.90)的不同细胞系中对溶酶体的标记表现出优异的能力。与一种商业上可用的 LysoTracker 不同,DNS-C 的斯托克斯位移高达 240nm(λ=330/570nm),比 LysoTracker 的~20nm(λ=573/595nm)大得多。更重要的是,DNS-C 在活细胞中进行长达 40 分钟的延时成像后,其荧光仍然保持高亮度,这意味着它具有出色的光稳定性,可用于长期跟踪。此外,DNS-C 还可以清晰地成像自噬体,这是一种关键的亚细胞区室,由溶酶体与自噬体融合形成。这些结果表明,我们的探针作为一种实时追踪溶酶体生理过程的强大工具具有广阔的应用前景。

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