研究目的
To develop a pH-independent rhodamine based spiro-ring platform for lysosomal imaging applications, specifically targeting HOCl with high sensitivity, selectivity, and fast response at lysosomal pH value.
研究成果
The developed thio-lactone rhodamine analogue HN4MS serves as an effective probe for HOCl sensing in lysosomes, demonstrating high sensitivity, selectivity, and fast response. Its successful application in visualizing exogenous and endogenous HOCl in living cells and tissues validates the strategy of using rhodamine spiro-ring derivatives for organelle-targetable imaging, offering potential for further development in bioimaging applications.
研究不足
The study's limitations include the probe's sensitivity to pH below 4.7 due to fluorescence quenching and reduced fluorescence responses at pH > 7.7 due to the pKa of HOCl/OCl? equilibrium. Additionally, the probe's performance in more complex biological systems or in vivo models was not extensively explored.
1：Experimental Design and Method Selection:
The study involved the design and synthesis of a thio-lactone rhodamine analogue HN4MS for HOCl sensing. The methodology included fluorescence spectroscopy and confocal microscopy for imaging applications.
2：Sample Selection and Data Sources:
HeLa cells and RAW
3：7 cells were used for imaging exogenous and endogenous HOCl, respectively. List of Experimental Equipment and Materials:
2 Instruments included a Bruker DRX-400 spectrometer for NMR, a Perkin elmer LS55 spectrofluorometer for fluorescence measurements, and an Olympus FV1000-IX81 laser scanning confocal microscope for imaging.
4：Experimental Procedures and Operational Workflow:
The synthesis of HN4M and HN4MS was followed by cytotoxicity studies, cell incubation, and fluorescence imaging to assess the probe's performance.
5：Data Analysis Methods:
Fluorescence quantum yields were determined using cresyl violet as a standard, and imaging data were analyzed for colocalization and intensity.
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