An azacyclo-localizing fluorescent probe for the specific labeling of lysosome and autolysosome
An azacyclo-localizing fluorescent probe for the specific labeling of lysosome and autolysosome
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用于特异性标记溶酶体和自溶酶体的氮杂环定位荧光探针
DOI：
10.1016/j.talanta.2020.120941
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发表时间：
2020
期刊：
影响因子：
6.1
通讯作者：
Zhang Zhongping
中科院分区：
文献类型：
--
作者：
Yang Linlin;Zhao Jun;Wang Jianping;Han Guangmei;Liu Bianhua;Zhang Wei;Fu Yao;Han Ming-Yong;Wang Zhenyang;Zhang Zhongping
了解溶酶体相关的生理学需要特异性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，我们报告了一个氮杂环修饰的荧光探针，具有较大的斯托克斯位移，良好的光稳定性和可忽略不计的细胞毒性，高度特异性标记的溶酶体和自体溶酶体在活细胞中。对母体染料丹磺酰基上不同氮杂环基团的探针进行了筛选，结果表明具有四个氮原子的丹磺酰基-环己基苯胺（DNS-C）具有最好的溶酶体定位能力。DNS-C作为一种通用的示踪剂，对不同细胞系的溶酶体标记具有良好的能力，重叠系数高（≥0.90）。与市售LysoTracker不同，DNS-C的斯托克斯位移高达240 nm（λex/em= 330/570 nm），远大于LysoTracker的斯托克斯位移~20 nm（λex/em= 573/595 nm）。更重要的是，DNS-C的荧光在活细胞中经过40 min的延时成像后仍然保持高亮度，这意味着其具有显著的长期跟踪光稳定性。此外，DNS-C还可以清晰地显示自噬过程中溶酶体与自噬体融合形成的自溶酶体，这是一个重要的亚细胞区室。这些结果表明，我们的探针作为一个强大的工具，实时跟踪溶酶体的生理过程的有前途的实用程序。
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 (λex/em= 330/570 nm), is much larger than that of LysoTracker ~20 nm (λex/em= 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.