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

Linlin Yang; Jun Zhao; Jianping Wang; Guangmei Han; Bianhua Liu; Wei Zhang; Yao Fu; Ming-Yong Han; Zhenyang Wang; Zhongping Zhang

doi:10.1016/j.talanta.2020.120941

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

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

Authors

Linlin Yang¹, Jun Zhao¹, Jianping Wang¹, Guangmei Han², Bianhua Liu¹, Wei Zhang³, Yao Fu⁴, Ming-Yong Han¹, Zhenyang Wang⁵, Zhongping Zhang⁶

Affiliations

¹ 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.
² 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. Electronic address: gmhan@iim.ac.cn.
³ School of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, 252059, China.
⁴ Department of Chemistry, University of Science and Technology of China, Hefei, 230026, China.
⁵ 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. Electronic address: zywang@iim.ac.cn.
⁶ 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.

PMID: 32456907
DOI: 10.1016/j.talanta.2020.120941

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_ex/em = 330/570 nm), is much larger than that of LysoTracker ~20 nm (λ_ex/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.

Keywords: Autolysosome; Azacyclo localization; Cell imaging; Fluorescent probe; Lysosome.

MeSH terms

Aza Compounds / chemical synthesis
Aza Compounds / chemistry*
Fluorescent Dyes / chemical synthesis
Fluorescent Dyes / chemistry*
HeLa Cells
Humans
Lysosomes / chemistry*
Optical Imaging
Tumor Cells, Cultured

Substances

Aza Compounds
Fluorescent Dyes