研究目的
To develop a light addressable potassium (K+) sensor for real-time monitoring of extracellular potassium concentration changes by stimulating living breast cancer cells.
研究成果
The developed light addressable potentiometric K+ sensor successfully demonstrated Nernstian response to [K+] in a light-defined spatial region and high selectivity for K+ in the presence of Na+. It enabled real-time monitoring of extracellular K+ concentration changes associated with living cells upon stimulation, with potential applications in analyte imaging and multiple ion sensing.
研究不足
The sensor's performance is influenced by the steric hindrance of ion-pairing with hydrophobic anions and the potential formation of irreversible ion adducts between Fc+ and Cl- or NO3-. The detection range is limited by the stability of the ionophore-analyte complex and the nature of co-extracting electrolyte.
1：Experimental Design and Method Selection:
The study utilized a light addressable potassium sensor based on a semiconducting silicon electrode with a thin ion-selective polymeric film. The sensor's operation involves localized activation of faradaic electrochemistry at the illuminated spot, enabling the oxidation of surface-bound ferrocene moieties to trigger K+ transfer.
2：Sample Selection and Data Sources:
Human breast cancer MCF-7 cells were used as model cells to test the sensor's applicability for real-time monitoring of extracellular [K+]. Intracellular K+ was monitored using Asante Potassium Green-2 (APG-2) dye.
3：List of Experimental Equipment and Materials:
The setup included a semiconducting silicon electrode, a thin ion-selective polymeric film containing valinomycin, a laser beam for localized illumination, and a microscope for imaging.
4：Experimental Procedures and Operational Workflow:
The sensor was tested by stimulating MCF-7 cells with valinomycin and monitoring the changes in extracellular [K+] electrochemically and intracellular [K+] via fluorescence microscopy.
5：Data Analysis Methods:
The voltammetric response was analyzed to determine the relationship between peak potential and K+ activity, demonstrating a Nernstian response.
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