ArticleACS chemical biology2015
Quantitative imaging of glutathione in live cells using a reversible reaction-based ratiometric fluorescent probe.
Article in ACS chemical biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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Who cites it
55 citing papers in PubMed, 190 citations in OpenAlex.
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- A locally activatable sensor for robust quantification of organellar glutathione.Nature chemistry · 2023Article
- Real-Time Monitoring of the Level and Activity of Intracellular Glutathione in Live Cells at Atomic Resolution byACS central science · 2023Article
- Thiol Quantification Using Colorimetric Thiol-Disulfide Exchange in Nonaqueous Solvents.ACS omega · 2023Article
- Light controlled reversible Michael addition of cysteine: a new tool for dynamic site-specific labeling of proteins.The Analyst · 2023Article
- Fluorescent Sensing of Glutathione and Related Bio-Applications.Biosensors · 2022Review
- Rational design of a dual-reactive probe for imaging the biogenesis of both HRSC chemical biology · 2022Article
- Fluorescent Probes and Mass Spectrometry-Based Methods to Quantify Thiols in Biological Systems.Antioxidants & redox signaling · 2022Review
- Discovery of a potent BTK and IKZF1/3 triple degrader through reversible covalent BTK PROTAC development.Current research in chemical biology · 2022Article
- Engineering a Smart Agent for Enhanced Immunotherapy Effect by Simultaneously Blocking PD-L1 and CTLA-4.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2021Article
- Seeking Illumination: The Path to Chemiluminescent 1,2-Dioxetanes for Quantitative Measurements andAccounts of chemical research · 2021Article
- Ratiometric Detection of Glutathione Based on Disulfide Linkage Rupture between a FRET Coumarin Donor and a Rhodamine Acceptor.Chembiochem : a European journal of chemical biology · 2021Article
- Thiol-Mediated Uptake.JACS Au · 2021Review
- Fluorescent Probes for Live Cell Thiol Detection.Molecules (Basel, Switzerland) · 2021Review
- Structure modulation on fluorescent probes for biothiols and the reversible imaging of glutathione in living cells.RSC advances · 2021Article
Corrections and comments
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Authors and funding
13 authors at 2 institutions in 1 country.
Funding
Abstract
Glutathione (GSH) plays an important role in maintaining redox homeostasis inside cells. Currently, there are no methods available to quantitatively assess the GSH concentration in live cells. Live cell fluorescence imaging revolutionized the field of cell biology and has become an indispensable tool in current biological studies. In order to minimize the disturbance to the biological system in live cell imaging, the probe concentration needs to be significantly lower than the analyte concentration. Because of this, any irreversible reaction-based GSH probe can only provide qualitative results within a short reaction time and will exhibit maximum response regardless of the GSH concentration if the reaction is completed. A reversible reaction-based probe with an appropriate equilibrium constant allows measurement of an analyte at much higher concentrations and, thus, is a prerequisite for GSH quantification inside cells. In this contribution, we report the first fluorescent probe-ThiolQuant Green (TQ Green)-for quantitative imaging of GSH in live cells. Due to the reversible nature of the reaction between the probe and GSH, we are able to quantify mM concentrations of GSH with TQ Green concentrations as low as 20 nM. Furthermore, the GSH concentrations measured using TQ Green in 3T3-L1, HeLa, HepG2, PANC-1, and PANC-28 cells are reproducible and well correlated with the values obtained from cell lysates. TQ Green imaging can also resolve the changes in GSH concentration in PANC-1 cells upon diethylmaleate (DEM) treatment. In addition, TQ Green can be conveniently applied in fluorescence activated cell sorting (FACS) to measure GSH level changes. Through this study, we not only demonstrate the importance of reaction reversibility in designing quantitative reaction-based fluorescent probes but also provide a practical tool to facilitate redox biology studies.
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Registered trials
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