研究目的
To develop a highly specific and ultrasensitive p-aminophenylether-based fluorescent probe for imaging native HOCl in live cells and zebrafish.
研究成果
The developed probe PAPE-HA exhibits high specificity, ultrasensitivity, and rapid response to HOCl, making it a promising tool for detecting native HOCl in live cells and zebrafish. This work not only provides a novel fluorescent probe for HOCl detection but also demonstrates the potential of p-aminophenylether moiety as an ideal recognition receptor for HOCl when combined with a suitable fluorophore.
研究不足
The study does not address the potential interference from other reactive oxygen species in more complex biological environments beyond the tested conditions. The probe's performance in vivo beyond zebrafish models was not explored.
1：Experimental Design and Method Selection:
The probe PAPE-HA was designed based on the 4-hydroxy-1,8-naphthalimide fluorophore with a p-aminophenylether group for specific detection of HOCl. The design rationale was to utilize the enhanced PET effect and inhibited ICT process for fluorescence quenching, which is reversed upon reaction with HOCl.
2：Sample Selection and Data Sources:
Live RAW
3：7 macrophage cells and zebrafish were used as biological samples to demonstrate the probe's capability in tracking native HOCl. List of Experimental Equipment and Materials:
2 Chemical reagents were obtained from commercial vendors. Instruments included LC-MS2010A for HRMS, Bruker AV-400 NMR spectrometer for NMR data, UV-3101PC spectrophotometer for absorption spectra, Horiba FluoroMax-4 spectrophotometer for fluorescence spectra, and Olympus FV1000-IX81 confocal fluorescence microscope for imaging.
4：Experimental Procedures and Operational Workflow:
The synthesis of probe PAPE-HA involved reacting N-butyl-4-chloro-1,8-naphthalimide with 4-aminophenol in acetonitrile. The probe's response to HOCl was tested in PBS solution, and its application in live cells and zebrafish was demonstrated through fluorescence imaging.
5：Data Analysis Methods:
Fluorescence intensities were measured and analyzed to determine the probe's sensitivity, selectivity, and response time to HOCl.
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