研究目的
To design a highly selective and sensitive fluorescent probe for simultaneously distinguishing and sequentially detecting H2S and various thiol species (Cys, Hcy, GSH) in solution and in live cells.
研究成果
The probe NCR effectively distinguishes and senses H2S, Cys/Hcy, and GSH with high selectivity and sensitivity in solution and live cells, demonstrating mitochondrial targeting and potential for biological applications in sulfur biology.
研究不足
The probe's performance is pH-dependent, with optimal response near neutral pH; potential interference from other biological species was tested but not exhaustively; application is limited to in vitro and cellular studies, not in vivo.
1：Experimental Design and Method Selection:
The probe NCR was designed with two fluorophores (coumarin and resorufin) and a nitrobenzoxadiazole (NBD) moiety, utilizing disulfide bond cleavage and FRET/ICT mechanisms for detection. Fluorescence spectroscopy and imaging techniques were employed.
2：Sample Selection and Data Sources:
Solutions of H2S, Cys, Hcy, and GSH in PBS buffer; HeLa cells for biological imaging.
3：List of Experimental Equipment and Materials:
Fluorescence spectrometer, cell culture equipment, MitoTracker Green for co-localization, N-ethylmaleimide (NEM) as a thiol scavenger.
4：Experimental Procedures and Operational Workflow:
Titration experiments with increasing amounts of analytes; cytotoxicity assessment via MTT assay; fluorescence imaging in cells with various treatments including NEM pre-treatment and addition of analytes.
5：Data Analysis Methods:
Fluorescence intensity measurements, calculation of detection limits, Pearson's correlation coefficient for co-localization, and statistical analysis of cytotoxicity.
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