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Updated: Jul 21, 2025
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Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator
Published on: October 20, 2021
2.9K
Quantification of glutathione with high throughput live-cell imaging
Biorxiv : the Preprint Server for Biology
|July 28, 2023
Summary
This study introduces RealThiol (RT), a novel fluorescent probe for accurately measuring glutathione (GSH) levels in living cells. RT enables real-time monitoring of GSH dynamics, crucial for understanding cellular redox state and disease.
Area of Science:
- Biochemistry
- Cell Biology
- Biomedical Engineering
Background:
- Redox reactions are fundamental to cellular processes, and disruptions in redox balance are linked to various human diseases.
- Glutathione (GSH) is a key intracellular antioxidant vital for maintaining redox homeostasis.
- Existing GSH probes rely on irreversible reactions, limiting real-time monitoring of GSH dynamics.
Approach:
- Developed the first reversible reaction-based fluorescent probe for glutathione.
- Utilized the second-generation probe, RealThiol (RT), for GSH measurement.
- Employed a high-throughput imaging system (Cytation™ 5) to assess RT's performance in living Hela cells treated with DL-Buthionine sulfoximine (BSO).
Key Points:
- RealThiol (RT) probe quantitatively monitors real-time GSH dynamics in living cells with high resolution.
- RT accurately detected changes in GSH levels in Hela cells following BSO inhibition of GSH synthesis.
- The probe demonstrated sensitivity and precision at a 2.0 µM concentration in high-throughput imaging.
Conclusions:
- The developed reversible fluorescent probe (RT) offers a sensitive and precise method for measuring intracellular GSH levels.
- RT facilitates real-time monitoring of GSH dynamics, essential for assessing cellular redox state.
- This technology has potential applications across various cell lines and high-throughput screening platforms.
