研究目的
The objective of this research is to develop a novel fluorescent probe both for extreme acidity detection and for extreme alkaline detection.
研究成果
Probe 1 is a effective ratiometric pH sensor based on TBET, capable of detecting extreme acidity and alkalinity with large emission shifts, reversibility, and applicability in cell imaging and vapor detection.
研究不足
The experiments were conducted in CH3CN/H2O (3:2, v/v) solution, which may not fully represent purely aqueous biological environments. The probe's performance in vivo or in more complex biological systems beyond Hela cells was not extensively explored. The pH range for linear response is specific (2.88-4.56 for acid, 9.82-13.78 for base), and performance at intermediate pH values (e.g., near neutral) is less emphasized.
1：Experimental Design and Method Selection:
The probe was designed based on through-bond energy transfer (TBET) using HBO as donor and Rhodamine derivative as acceptor. Synthesis involved a simple condensation reaction. Optical properties were studied using absorption and fluorescence spectroscopy in CH3CN/H2O (3:2, v/v) solution at different pH values. 1H NMR titration was used to investigate structural changes. Cell imaging was performed using fluorescence microscopy.
2：Sample Selection and Data Sources:
Probe 1 was synthesized and characterized. Hela cells were used for biological studies. pH buffers were prepared for spectral measurements.
3：List of Experimental Equipment and Materials:
Spectrophotometer for absorption and fluorescence spectra, NMR spectrometer for structural analysis, fluorescence inversion microscope (Zeiss Axio Observer A1) for cell imaging, filter papers for test strips, CH3CN, H2O, DMSO-d6, NaOH, HCl, metal ions for selectivity tests.
4：Experimental Procedures and Operational Workflow:
Probe 1 (10 μM) was dissolved in CH3CN/H2O (3:2, v/v) and pH adjusted with HCl/NaOH. Spectra recorded at various pH. Reversibility tested by cycling pH. Photostability monitored over 120 min. Test papers prepared by immersion in probe solution and exposed to HCl/NH3 vapor. Cell imaging done after incubating Hela cells with probe at different pH.
5：Data Analysis Methods:
Fluorescence intensity ratios (I593/I422 and I555/I422) calculated for ratiometric detection. Linear ranges determined from ratio vs. pH plots. Selectivity assessed by comparing responses to metal ions. Cytotoxicity evaluated using CCK8 assay.
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