Real-time monitoring of endogenous cysteine levels in living cells using a CD-based ratiometric fluorescent nanoprobe
- 465 Downloads
A simple and readily available fluorescent probe is needed for the real-time monitoring of endogenous cysteine (Cys) levels in living cells, as such a probe could be used to study the role of Cys in related diseases. Herein, we report the first fluorescent probe based on carbon dots (CDs-FITA) for the selective and ratiometric imaging of endogenous Cys in live cells. In this ratiometric fluorescent probe, a fluorescein derivative (FITA) that recognizes Cys is covalently linked to the surfaces of carbon dots (CDs); employing CDs greatly improves the water solubility of the probe. Acrylate on FITA is selectively cleaved by Cys in aqueous solution under mild conditions, leading to a dramatic increase in the fluorescence from fluorescein. The probe therefore allows the highly selective ratiometric fluorescent detection of Cys even in the presence of various interferents. The as-prepared CDs-FITA showed excellent performance when applied to detect Cys in blood serum. In addition, due to its negligible cytotoxicity, the CDs-FITA can also be utilized for the real-time monitoring of endogenous cysteine (Cys) levels in living cells.
KeywordsCarbon dots Fluorescein Ratiometric fluorescence Cys Living cell imaging
We gratefully acknowledge the financial support provided by NSFC (project nos. 51603067, 51773056, and 51373002), the Hunan Provincial Natural Science Foundation of China (project nos. 2018JJ3143 and 2016JJ5005), the Open Project Program of State Key Laboratory of Chemo/Biosensing and Chemometrics (project nos. 2016019, 2013008), and the China Postdoctoral Science Foundation (project no. 2017 M622571).
Compliance with ethical standards
Conflicts of interest
There are no conflicts of interest to declare.
- 8.Jung HS, Han JH, Pradhan T, Kim S, Lee SW, Sessler JL, et al. A cysteine-selective fluorescent probe for the cellular detection of cysteine. Biomaterials. 2012;33(3):945–53.Google Scholar
- 9.Chen W, Xu S, Day JJ, Wang D, Xian M. A general strategy for development of near-infrared fluorescent probes for bioimaging. Angew Chem Int Ed. 2017;56(52):16611–5.Google Scholar
- 18.Liu X, Tian H, Yang L, Su Y, Guo M, Song X. An ESIPT-based fluorescent probe for sensitive and selective detection of Cys/Hcy over GSH with a red emission and a large Stokes shift. Tetrahedron Lett. 2015;58:3209–13.Google Scholar
- 23.Zhang P, Nie X, Gao M, Zeng F, Qin A, Wu S. Tang BZ. A highly selective fluorescent nanoprobe based on AIE and ESIPT for imaging hydrogen sulfide in live cells and zebrafish. Mater Chem Front. 2017;1(5):838–45.Google Scholar
- 25.Chen W, Rosser EW, Matsunaga T, Pacheco A, Akaike T, Xian M. The development of fluorescent probes for visualizing intracellular hydrogen polysulfides. Angew Chem Int Ed. 2015;54(47):13961–5.Google Scholar
- 26.Yang X, Guo Y, Strongin RM. Conjugate addition/cyclization sequence enables selective and simultaneous fluorescence detection of cysteine and homocysteine. Angew Chem Int Ed. 2011;50:10690–3.Google Scholar
- 30.Wang J, Li B, Zhao W, Zhang X, Luo X, Corkins ME, Cole SL, Wang C, Xiao Y, Bi X, Pang Y, McElroy CA, Bird AJ, Dong Y. Two-photon near infrared fluorescent turn-on probe toward cysteine and its imaging applications. ACS Sens. 2016;1(7):882–7.Google Scholar
- 32.Qi Y, Huang Y, Li B, Zeng F, Wu S. Real-time monitoring of endogenous cysteine levels in vivo by near-infrared turn-on fluorescent probe with large Stokes shift. Anal Chem. 2018;90(1):1014–20.Google Scholar
- 37.Ouadahi K, Sbargoud K, Allard E, Larpent C. FRET-mediated pH-responsive dual fluorescent nanoparticles prepared via click chemistry. Nano. 2012;4(3):727–32.Google Scholar
- 52.Zhao F, Qian J, Quan F, Wu C, Zheng Y, Zhou L. Aconitic acid derived carbon dots as recyclable “on-off-on” fluorescent nanoprobes for sensitive detection of mercury(II) ions, cysteine and cellular imaging. RSC Adv. 2017;7(40):44178–85.Google Scholar
- 53.Yan F, Shi D, Zheng T, Yun K, Zhou X, Chen L. Carbon dots as nanosensor for sensitive and selective detection of Hg2+ and L-cysteine by means of fluorescence “off-on” switching. Sensors Actuators B Chem. 2016;224:926–35.Google Scholar
- 55.Huang H, Weng Y, Zheng L, Yao B, Weng W, Lin X. Nitrogen-doped carbon quantum dots as fluorescent probe for “off-on” detection of mercury ions, L-cysteine and iodide ions. J Colloid Interface Sci. 2017;506:373–8.Google Scholar
- 61.Zhang P, Wang H, Hong Y, Yu M, Zeng R, Long Y, et al. Selective visualization of endogenous hypochlorous acid in zebrafish during lipopolysaccharide-induced acute liver injury using a polymer micelles-based ratiometric fluorescent probe. Biosens Bioelectron. 2018;99:318–24.CrossRefPubMedGoogle Scholar
- 66.Bhattacharya S, Sarkar R, Chakraborty B, Porgador A, Jelinek R. Nitric oxide sensing through azo-dye formation on carbon dots. ACS Sens. 2017;2(8):1215–24.Google Scholar