研究目的
To develop a novel mitochondria-targeting near-infrared fluorescent probe for the detection and imaging of hydrogen sulfide (H2S) in living cells, addressing the need for sensitive, rapid, and reliable detection techniques for endogenous H2S in biological systems.
研究成果
The probe Mito-NIR-SH demonstrates superior chemical stability, fast response, high sensitivity and selectivity for H2S detection, and effective mitochondrial targeting. It successfully images both exogenous and endogenous H2S in living cells, making it a valuable tool for studying H2S-related physiological and pathological processes, with potential for further applications in biomedical research.
研究不足
The probe's performance was tested primarily in HeLa cells and under controlled laboratory conditions; its applicability to other cell types or in vivo systems may require further validation. The response time, while fast, could potentially be optimized for real-time applications. The detection limit, though low, might not capture the lowest endogenous levels in all biological contexts.
1：Experimental Design and Method Selection:
The probe Mito-NIR-SH was designed based on an intramolecular charge transfer (ICT) fluorophore (CS-OH) with a 2,4-dinitrophenyl (DNB) group as the H2S response site. The synthesis involved a three-step reaction, and characterization was done using NMR and mass spectrometry. Spectroscopic properties were evaluated using UV-vis and fluorescence spectroscopy.
2：Sample Selection and Data Sources:
HeLa cells were used for cytotoxicity and imaging studies, sourced from XiangYa School of Medicine, Central South University. Chemical reagents were commercially available.
3：List of Experimental Equipment and Materials:
Equipment included BRUKER NMR spectrometers, Shimadzu UV-2550 spectrophotometer, Hitachi F-4600 fluorometer, Olympus FV1000-MPE confocal microscope, Agilent HPLC system, Leici PHS-3C pH meter, and LCMS-IT-TOF mass spectrometer. Materials included DMSO, NaHS, PBS buffer, and various ions and molecules for selectivity tests.
4：Experimental Procedures and Operational Workflow:
Synthesis of compounds, preparation of stock solutions, absorption and fluorescence spectra measurements, selectivity and competition studies, cytotoxicity assays using MTT, and cell culture and imaging experiments with confocal microscopy.
5：Data Analysis Methods:
Fluorescence intensity measurements, linear regression for detection limits, co-localization analysis for mitochondrial targeting, and statistical analysis for cytotoxicity.
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