Theranostics 2020; 10(5):2130-2140. doi:10.7150/thno.39673 This issue

Research Paper

Glutathione-responsive disassembly of disulfide dicyanine for tumor imaging with reduction in background signal intensity

Shanyan Mo1,2*, Xiaoting Zhang1*, Sadaf Hameed1*, Yiming Zhou1, Zhifei Dai1✉

1. Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China
2. College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
* These authors contributed equally to this work.

This is an open access article distributed under the terms of the Creative Commons Attribution License ( See for full terms and conditions.
Mo S, Zhang X, Hameed S, Zhou Y, Dai Z. Glutathione-responsive disassembly of disulfide dicyanine for tumor imaging with reduction in background signal intensity. Theranostics 2020; 10(5):2130-2140. doi:10.7150/thno.39673. Available from

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Graphic abstract

Near-infrared (NIR) fluorescence imaging has been proved as an effective modality in identifying the tumor border and distinguishing the tumor cells from healthy tissue during the oncological surgery. Developing NIR fluorescent probes with high tumor to background (T/B) signal is essential for the complete debulking of the tumor, which will prolong the survival rate of tumor patients. However, the nonspecific binding and “always-on” properties of the conventional fluorescent probes leads to high background signals and poor specificity.

Method: To address this problem, glutathione (GSH)-responsive, two disulfide-bonded dicyanine dyes (ss-diCy5 and ss-diNH800CW) were synthesized. As synthesized dyes are quenched under normal physiological conditions, however, once reached to the tumor site, these dyes are capable of emitting strong fluorescence signals primarily because of the cleavage of the disulfide bond in the tumor microenvironment with high GSH concentration. Besides, the GSH-responsive behavior of these dyes was monitored using the UV-vis and fluorescence spectroscopy. The diagnostic accuracy of the aforementioned dyes was also tested both in tumor cells and 4T1-bearing mice.

Results: The fluorescence signal intensity of disulfide dicyanine dyes was quenched up to 89% compared to the mono cyanine dyes, thus providing a very low fluorescence background. However, when the disulfide dicyanine dye reaches the tumor site, the dicyanine is cleaved by GSH into two mono-dyes with high fluorescence strength, thus producing strong fluorescent signals upon excitation. The fluorescent signal of the dicyanine was enhanced by up to 27-fold after interacting with the GSH solution. In vivo xenografts tumor studies further revealed that the fluorescence signals of aforementioned dyes can be quickly recovered in the solid tumor.

Conclusion: In summary, the disulfide dicyanines dyes can provide a promising platform for specific tumor-activatable fluorescence imaging with improved T/B value.

Keywords: Fluorescence imaging, molecular probe, tumor response, disulfide dicyanine, fluorescence quenching