研究目的
To develop a novel fluorescent sensor for detecting Zn2+ with high selectivity and sensitivity, and to apply it for imaging Zn2+ in living cells.
研究成果
The developed fluorescent sensor exhibits high selectivity and sensitivity for Zn2+ detection in aqueous solutions, with a 34-fold fluorescence enhancement and a low detection limit of 1.72×10-6 mol·L-1. It successfully images Zn2+ in living HeLa cells, demonstrating good membrane permeability and potential for biological applications. This work advances the field of zinc sensing with improved performance over previous sensors.
研究不足
The experiments were conducted under specific pH conditions (pH=7.4) using HEPES buffer, which may not fully represent all biological environments. The sensor's performance in complex matrices or other cell types was not explored, and potential interferences from other ions or cellular components were not thoroughly investigated. Optimization of synthesis and application in vivo could be areas for future improvement.
1：Experimental Design and Method Selection:
The study involved designing and synthesizing a 4-amino-1,8-naphthalimide-based fluorescent sensor with iminodiacetic acid as a receptor, utilizing a PET (Photo-induced Electron Transfer) mechanism for Zn2+ detection. Fluorescence spectroscopy was employed to evaluate selectivity and sensitivity in aqueous solutions under physiological pH conditions. Molecular modeling using DFT calculations was used to explore binding properties.
2：Sample Selection and Data Sources:
The sensor was synthesized as described in the Supporting Information. Fluorescence properties were tested in HEPES buffer (20 mM, pH=7.4) with various metal ions, including Zn2+, to assess selectivity and sensitivity. Living HeLa cells were used for imaging experiments.
3：4) with various metal ions, including Zn2+, to assess selectivity and sensitivity. Living HeLa cells were used for imaging experiments.
List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: Equipment included a SHIMADZU IR Affinity-1 FT-IR spectrophotometer for IR measurements, and unspecified fluorescence spectrometers for emission studies. Materials included HEPES buffer, Zn2+ ions, and the synthesized sensor compound.
4：Experimental Procedures and Operational Workflow:
The sensor was dissolved in HEPES buffer, and fluorescence spectra were recorded with excitation at 470 nm. Titration experiments with increasing Zn2+ concentrations were performed to measure sensitivity. Job's plot and mass spectrometry (MS) were used to determine binding stoichiometry. For cell imaging, HeLa cells were incubated with the sensor and Zn2+, followed by fluorescence microscopy.
5：Data Analysis Methods:
Fluorescence intensity changes were analyzed to calculate enhancement factors and detection limits using standard deviation and linear regression. DFT calculations (B3LYP/6-311G level with Gaussian 2009) optimized the sensor-Zn2+ complex conformation. Statistical analysis included R-squared values for linear fits.
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