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Live-Cell Derivatization Enables High-Throughput Analysis of Thiols with CyESI-MS
Kexin Xu1, Jinlei Yang1, Siyuan Pan1
1Department of Chemistry, Tsinghua University, Beijing 100084, China.
Analytical Chemistry
|May 5, 2026
Summary
This study introduces a new method for detecting thiol metabolites in single cells using mass cytometry. This advance improves the analysis of redox regulation and disease, aiding cancer research.
Area of Science:
- Biochemistry
- Cell Biology
- Analytical Chemistry
Background:
- Detecting single-cell thiol metabolites is vital for understanding redox regulation and disease.
- Low abundance and signal suppression hinder thiol detection in mass spectrometry (MS).
Purpose of the Study:
- To develop a sensitive, high-throughput method for live-cell thiol metabolite analysis.
- To overcome MS signal limitations for low-abundance metabolites.
Main Methods:
- Developed a live-cell derivatization strategy using Tetramethylrhodamine-5-maleimide (TAMRA-5-maleimide).
- Coupled derivatization with label-free mass cytometry (CyESI-MS) for analysis.
- Utilized TAMRA-5-maleimide for thiol specificity, signal enhancement, and biocompatibility.
Main Results:
- Successfully detected five thiol metabolites (GSH, Cys, Cys-Gly, hCys, NCys) in single cells.
- Demonstrated method reliability in various cell-based applications.
- Achieved sensitive, high-throughput analysis of low-abundance thiol metabolites.
Conclusions:
- The TAMRA-5-maleimide derivatization and CyESI-MS strategy enables sensitive single-cell thiol detection.
- This approach enhances the study of redox biology and cancer pathogenesis.
- Expands single-cell metabolite coverage for future research.
