Skip to main content

Advertisement

Log in

Rational design of a near-infrared dual-emission fluorescent probe for ratiometric imaging of glutathione in cells

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

Sensors for which the output signal is an intensity change for a single-emission peak are easily disturbed by many factors, such as the stability of the instrument, intensity of the excitation light, and biological background. However, for ratiometric fluorescence sensors, the output signal is a change in the intensity ratio of two or more emission peaks. The fluorescence intensity of these emission peaks is similarly affected by external factors; thus, these sensors have the ability to self-correct, which can greatly improve the accuracy and reliability of the detection results. To accurately image glutathione (GSH) in cells, gold nanoclusters (AuNCs) with intrinsic double emission at wavelengths of 606 nm and 794 nm were synthesized from chloroauric acid. With the emission peak at 606 nm as the recognition signal and the emission peak at 794 nm as the reference signal, a near-infrared dual-emission ratio fluorescence sensing platform was constructed to accurately detect changes in the GSH concentration in cells. In vitro and in vivo analyses showed that the ratiometric fluorescent probe specifically detects GSH and enables ultrasensitive imaging, providing a new platform for the accurate detection of active small molecules.

Graphical Abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data availability

The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request.

References

  1. Liu J, Bao C, Zhong X, Zhao C, Zhu L (2010) Highly selective detection of glutathione using a quantum-dot-based off-on fluorescent probe. Chem Commun 46:2971–2973

    Article  CAS  Google Scholar 

  2. Yang XF, Huang Q, Zhong Y, Li Z, Li H, Lowry M, Strongin ERM (2014) A dual emission fluorescent probe enables simultaneous detection of glutathione and cysteine/homocysteine. Chem Sci 5:2177–2183

    Article  CAS  PubMed  Google Scholar 

  3. He L, Xu Q, Liu Y, Wei H, Tang Y, Lin W (2015) Coumarin-based turn-on fluorescence probe for specific detection of glutathione over cysteine and homocysteine. ACS Appl Mater Interfaces 7:12809–12813

    Article  CAS  PubMed  Google Scholar 

  4. Huang Y, Zhou J, Feng H, Zheng J, Ma HM, Liu W, Tang C, Ao H, Zhao M, Qian Z (2016) A dual-channel fluorescent chemosensor for discriminative detection of glutathione based on functionalized carbon quantum dots. Biosens Bioelectron 86:748–755

    Article  CAS  PubMed  Google Scholar 

  5. Jung HS, Chen X, Kim JS, Yoon J (2013) Recent progress in luminescent and colorimetric chemosensors for detection of thiols. Chem Soc Rev 42:6019–6031

    Article  CAS  PubMed  Google Scholar 

  6. Yin C, Tang Y, Li X, Yang Z, Li J, Li X, Huang W, Fan Q (2018) A single composition architecture-based nanoprobe for ratiometric photoacoustic imaging of glutathione (GSH) in living mice. Small 14:1703400

    Article  Google Scholar 

  7. Sekhar RV, McKay SV, Patel SG, Guthikonda AP, Reddy VT, Balasubramanyam A, Jahoor F (2011) Glutathione synthesis is diminished in patients with uncontrolled diabetes and restored by dietary supplementation with cysteine and glycine. Diabetes Care 34:162–167

    Article  CAS  PubMed  Google Scholar 

  8. Wang S, Zhang L, Zhao J, He M, Huang Y, Zhao S (2021) A tumor microenvironment–induced absorption red-shifted polymer nanoparticle for simultaneously activated photoacoustic imaging and photothermal therapy. Sci Adv 7:eabe3588

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  9. Li F, Liu J, Hu Y, Deng N, He J (2018) An ultrasensitive label-free colorimetric assay for glutathione based on Ag+ regulated autocatalytic oxidation of o-phenylenediamine. Talanta 186:330–336

    Article  CAS  PubMed  Google Scholar 

  10. Townsend DM, Tew KD, Tapiero H (2003) The importance of glutathione in human disease. Biomed Pharmacother 57:145–155

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Janeš L, Lisjak K, Vanzo A (2010) Determination of glutathione content in grape juice and wine by high-performance liquid chromatography with fluorescence detection. Anal Chim Acta 674:239–242

    Article  PubMed  Google Scholar 

  12. Yang Q, Krautmacher C, Schilling D, Pittelkow MR, Naylor S (2010) Simultaneous analysis of oxidized and reduced glutathione in cell extracts by capillary zone electrophoresis. Biomed Chromatogr 16:224–228

    Article  CAS  Google Scholar 

  13. Zhao L, Liu Z, Miao Y, Zhang C (2016) Selective electrochemical determination of glutathione from the leakage of intracellular GSH contents in HeLa cells following doxorubicin-induced cell apoptosis. Electrochim Acta 206:86–98

    Article  CAS  Google Scholar 

  14. Saha A, Jana NR (2013) Detection of cellular glutathione and oxidized glutathione using magnetic-plasmonic nanocomposite-based “turn-off” surface enhanced Raman scattering. Anal Chem 85:9221–9228

    Article  CAS  PubMed  Google Scholar 

  15. Huang GG, Han XX, Hossain MK, Ozaki Y (2009) Development of a heat-induced surface-enhanced Raman scattering sensing method for rapid detection of glutathione in aqueous solutions. Anal Chem 81:5881–5888

    Article  CAS  PubMed  Google Scholar 

  16. Lv P, Xu Y, Liu Z, Li G, Ye B (2020) Carbon dots doped lanthanide coordination polymers as dual-function fluorescent probe for ratio sensing Fe and ascorbic acid. Microchem J 152:104255–104261

    Article  CAS  Google Scholar 

  17. Huang S, Yao J, Chu X, Liu Y, Xiao Q, Zhang Y (2019) One-step facile synthesis of nitrogen-doped carbon dots: a ratiometric fluorescent probe for evaluation of acetylcholinesterase activity and detection of organophosphorus pesticides in tap water and food. J Agric Food Chem 67:11244–11255

    Article  CAS  PubMed  Google Scholar 

  18. Liu JW, Luo Y, Wang YM, Duan LY, Jiang JH, Yu RQ (2016) Graphitic carbon nitride nanosheets-based ratiometric fluorescent probe for highly sensitive detection of H2O2 and glucose. ACS Appl Mater Interfaces 8:33439–33445

    Article  CAS  PubMed  Google Scholar 

  19. Cen Y, Wu YM, Kong XJ, Wu S, Yu RQ, Chu X (2014) Phospholipid-modified upconversion nanoprobe for ratiometric fluorescence detection and imaging of phospholipase D in cell lysate and in living cells. Anal Chem 86:7119–7127

    Article  CAS  PubMed  Google Scholar 

  20. Li H, Liu Y, Huang B, Zhang C, Wang Z, She W, Liu Y, Jiang P (2022) Highly efficient GSH-responsive “off−on” NIR-II fluorescent fenton nanocatalyst for multimodal imaging-guided photothermal/chemodynamic synergistic cancer therapy. Anal Chem 94:10470–10478

    Article  CAS  PubMed  Google Scholar 

  21. Lin X, Zhu R, Hong Z, Zhang X, Chen S, Song J, Yang H (2021) GSH-responsive radiosensitizers with deep penetration ability for multimodal imaging-guided synergistic radio-chemodynamic cancer therapy. Adv Funct Mater 31:2101278

  22. Su X, Zhuang W, Yu T, He H, Ma B, Chen L, Yang L, Li G, Wang Y (2020) ROS and GSH dual-responsive gem prodrug micelles for ROS-triggered fluorescence turn on bioimaging and cancer therapy. Adv Mater Interfaces 7:2000294

  23. He YS, Pan CG, Cao HX, Yue MZ, Wang L, Liang GX (2018) Highly sensitive and selective dual-emission ratiometric fluorescence detection of dopamine based on carbon dots-gold nanoclusters hybrid. Sensor Actuat B-Chem 265:371–377

    Article  CAS  Google Scholar 

  24. Xiao H, Li P, Zhang W, Tang B (2016) An ultrasensitive near-infrared ratiometric fluorescent probe for imaging mitochondrial polarity in live cells and in vivo. Chem Sci 7:1588–1593

    Article  CAS  PubMed  Google Scholar 

  25. Zhu H, Yu T, Xu H, Zhang K, Jiang H, Zhang Z, Wang Z, Wang S (2014) Fluorescent nanohybrid of gold nanoclusters and quantum dots for visual determination of lead ions. ACS Appl Mater Interfaces 6:21461–21467

    Article  CAS  PubMed  Google Scholar 

  26. Du F, Min Y, Zeng F, Yu C, Wu S (2014) A targeted and FRET-based ratiometric fluorescent nanoprobe for imaging mitochondrial hydrogen peroxide in living cells. Small 10:964–972

    Article  CAS  PubMed  Google Scholar 

  27. Ling X, Gong D, Shi W, Xu Z, Han W, Lan G, Li Y, Qin W, Lin W (2021) Nanoscale metal-organic layers detect mitochondrial dysregulation and chemoresistance via ratiometric sensing of glutathione and pH. J Am Chem Soc 143:1284–1289

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Shang L, Dong S, Nienhaus G (2011) Ultra-small fluorescent metal nanoclusters: synthesis and biological applications. Nano Today 6:401–418

    Article  CAS  Google Scholar 

  29. Chen LY, Wang CW, Yuan Z, Chang HT (2015) Fluorescent gold nanoclusters: recent advances in sensing and imaging. Anal Chem 87:216–229

    Article  CAS  PubMed  Google Scholar 

  30. Luo Z, Yuan X, Yu Y, Zhang Q, Leong DT, Lee JY, Xie J (2012) From aggregation-induced emission of Au(I)-thiolate complexes to ultrabright Au(0)@Au(I)-thiolate core-shell nanoclusters. J Am Chem Soc 134:16662–16670

    Article  CAS  PubMed  Google Scholar 

  31. Negishi Y, Nobusada K, Tsukuda T (2005) Glutathione-protected gold clusters revisited: bridging the gap between gold(I)-thiolate complexes and thiolate-protected gold nanocrystals. J Am Chem Soc 127:5261–5270

    Article  CAS  PubMed  Google Scholar 

  32. Li W, Wang X, Jiang T, Ma X, Tian H (2020) One-pot synthesis of β-cyclodextrin modified Au nanoclusters with near-infrared emission. Chem Commun 56:5580–5583

    Article  CAS  Google Scholar 

  33. Xie J, Zheng Y, Ying JY (2010) Highly selective and ultrasensitive detection of Hg2+ based on fluorescence quenching of Au nanoclusters by Hg2+-Au+ interactions. Chem Commun 46:961–963

    Article  CAS  Google Scholar 

  34. Cai Y, Yan L, Liu G, Yuan H, Xiao D (2013) In-situ synthesis of fluorescent gold nanoclusters with electrospun fibrous membrane and application on Hg (II) sensing. Biosens Bioelectron 41:875–879

    Article  CAS  PubMed  Google Scholar 

  35. Zhang Y, Jiang J, Li M, Gao P, Shi L, Zhang G, Dong C, Shuang S (2017) Bright far-red/near-infrared gold nanoclusters for highly selective and ultra-sensitive detection of Hg2+. Sensor Actuat B-Chem 238:683–692

    Article  CAS  Google Scholar 

  36. Wu F, Tong C (2019) Nitrogen- and sulfur-codoped carbon dots for highly selective and sensitive fluorescent detection of Hg2+ ions and sulfide in environmental water samples. J Agric Food Chem 67:2794–2794

    Article  CAS  PubMed  Google Scholar 

  37. Stricks W, Kolthoff IM (1953) Reactions between mercuric mercury and cysteine and glutathione. Apparent dissociation constants, heats and entropies of formation of various forms of mercuric mercapto-cysteine and –glutathione. J Am Chem Soc 75:5673–5681

    Article  CAS  Google Scholar 

  38. Cheesman BV, Arnold AP, Rabenstein DL (1988) Nuclear magnetic resonance studies of the solution chemistry of metal complexes. 25. Hg(thiol)3 complexes and Hg(II)-thiol ligand exchange kinetics. J Am Chem Soc 110:6359–6364

    Article  CAS  Google Scholar 

  39. Yue Y, Huo F, Yue P, Meng X, Salamanca JC, Escobedo JO, Strongin RM, Yin C (2018) In situ lysosomal cysteine-specific targeting and imaging during dexamethasone induced apoptosis. Anal Chem 90:7018–7024

    Article  CAS  PubMed  Google Scholar 

  40. Zu F, Yan F, Bai Z, Xu J, Wang Y, Huang Y, Zhou X (2017) The quenching of the fluorescence of carbon dots: a review on mechanisms and applications. Microchim Acta 184(7):1899–1914

    Article  CAS  Google Scholar 

  41. Zhang Y, Shao X, Wang Y, Pan F, Kang R, Peng F, Huang Z, Zhang W, Zhao W (2015) Dual emission channels for sensitive discrimination of Cys/Hcy and GSH in plasma and cells. Chem Commun 51:4245–4248

    Article  CAS  Google Scholar 

  42. Guo T, Chen X, Qu W, Yang B, Tian R, Geng Z, Wang Z (2022) Red and Near-infrared fluorescent probe for distinguishing cysteine and homocysteine through single-wavelength excitation with distinctly dual emissions. Anal Chem 94:5006–5013

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported by the National Natural Science Foundations of China (Nos. 22346021, 22064018, 51963021), the Natural Science Foundation of Guangxi Province of China (2020GXNSFAA159072), and the PhD Research Startup Program of Yulin Normal University (G2022ZK21, G2023ZK04), as well as the China University Students Innovative Project (202310606029).

Author information

Authors and Affiliations

Authors

Contributions

SW and JX designed the experiments. SW, ZL, MF, WH, and QL designed and synthesized the probe and studied the viability and cell imaging. RW and ZL assisted with the cell imaging data analysis and the manuscript drafting. All authors analyzed and discussed the data and contributed to the writing of the manuscript. All authors approved the final version of the manuscript.

Corresponding authors

Correspondence to Jiayao Xu, Rong Wang or Zhihui Luo.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 908 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, S., Li, Z., Xu, J. et al. Rational design of a near-infrared dual-emission fluorescent probe for ratiometric imaging of glutathione in cells. Microchim Acta 191, 92 (2024). https://doi.org/10.1007/s00604-024-06179-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-024-06179-6

Keywords

Navigation