Recognition of ClO－ and Cellular Imaging with Xanthene-based Fluorescent Probes

doi:10.19894/j.issn.1000-0518.220071

Chinese Journal of Applied Chemistry ›› 2022, Vol. 39 ›› Issue (12): 1903-1911.DOI: 10.19894/j.issn.1000-0518.220071

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Recognition of ClO^－ and Cellular Imaging with Xanthene-based Fluorescent Probes

Si-Wei YU¹^,², Liang-Peng WANG¹^,², Ri-Zhe JIN¹, Chuan-Qing KANG¹^,²()

^1.Laboratory of Polymer Composite and Engineering，Changchun Institute of Applied Chemistry，Chinese Academy of Sciences，Changchun 130022，China
^2.University of Science and Technology of China，Hefei 230022，China

Received:2022-03-15 Accepted:2022-07-23 Published:2022-12-01 Online:2022-12-13
Contact: Chuan-Qing KANG
About author:kangcq@ciac.ac.cn
Supported by:
the Science and Technology Project of Jilin Province, China(20210401164YY)

1. .jpg(7770KB)

Abstract

Abstract:

Selective recognition of hypochlorite ion （ClO^－） in intracellular biological environment is important for research and application in cellular physiology and pathology. A novel fluorescent molecule， 6-bis（dimethylamino）-9-（1-（3-methyl-benzothiazole）-trimethyl）-xanthene （R-1）， is designed and synthesized with a combination of dimethylamino-substituted xanthene and benzothiazole segments through continuous carbon-carbon double bonds. In the presence of ClO^－， the fluorescence emission of R-1 at 600 nm is significantly enhanced （the quantum yield is as high as 20.3%）， and other reactive oxygen species do not affect the recognition of ClO^－. Fluorescence imaging experiments demonstrate that probe R-1 can detect ClO^－ in human lung adenocarinoma （A549） cells and emit red fluorescence， indicating the potential application of R-1 in ClO^－ biological imaging.

Key words: Fluorescent probe, Xanthene, Benzothiazole, Hypochlorite ion, Fluorescence imaging

CLC Number:

O625.6

Si-Wei YU, Liang-Peng WANG, Ri-Zhe JIN, Chuan-Qing KANG. Recognition of ClO^－ and Cellular Imaging with Xanthene-based Fluorescent Probes[J]. Chinese Journal of Applied Chemistry, 2022, 39(12): 1903-1911.

Figures/Tables 9

Fig.1 Synthetic route of probe R-1

Fig.2 （A） UV-Vis absorption spectra of probe R-1 with different concentrations of sodium hypochlorite in methanol/water；（B） A linear plot of the UV absorption at 650 nm versus concentrations of ClO－

Fig.3 Fluorescence spectra of probe R-1 （10 μmol/L） in response to various concentrations of ClO－（0～20 μmol/L） in methanol/water solutions（A） Concentration of ClO－ increased from 0 μmol/L to 10 μmol/L；（B） Concentration of ClO－ increased from 10 μmol/L to 20 μmol/L； Inset： linear plots of the fluorescence intensities of probe R-1 at 600 nm versus concentrations of ClO－

Fig.4 Fluorescence lifetime of probe R-1 （10 μmol/L） in response to ClO－（10 μmol/L） in methanol/water solutions

Fig.5 Selective recognition of probe R-1 to ClO－ in methanol/water solution（A） Fluorescence spectra of probe R-1 （10 μmol/L） in the presence of ions and reactive oxygen species in methanol/water；（B） Fluorescence intensity of probe R-1 at emission wavelength of 600 nm in the presence of ions and reactive oxygen species；（C） Color of probe R-1 in the presence of ions and reactive oxygen species； Concentration of ClO－ was 10 μmol/L， others were 100 μmol/L

Fig.6 Mass spectra of probe R-1 before （A） and after （B） reaction with sodium hypochlorite

Fig.7 Proposed response mechanism of probe R-1 to hypochlorite

Fig.8 Cytotoxicity of probe R-1（A） and LPS（B） on A549 cells

Fig.9 Fluorescence imaging of probe R-1 in A549 cell line（scale bar： 20 μm）First row： A549 cells stained with Hoechst 33342； Second row： A549 cells stained with probe R-1 and Hoechst 33342； Third row： A549 cells stained with probe R-1 and Hoechst 33342 after incubated with LPS （1 μg/mL）； Fourth row： A549 cells were stained with probe R-1 and Hoechst 33342 after incubated with sodium hypochlorite aqueous solution （5 μmol/L）

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Recognition of ClO^－ and Cellular Imaging with Xanthene-based Fluorescent Probes

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