研究目的
To implement steady-state bright electrochemiluminescence (ECL) at single semiconductive titanium dioxide (TiO2) nanoparticles for sensing the local efflux from single living cells.
研究成果
The study successfully demonstrated steady-state bright ECL at single semiconductive TiO2 nanoparticles for local sensing of hydrogen peroxide efflux from single living cells. This approach provides a novel method for subcellular electroanalysis using single nanoparticles, with potential applications in studying cellular activities and molecular distributions.
研究不足
The spatial resolution of the ECL image is restricted to ~2 μm due to the diffusion of radicals during the ECL process. The quantification of hydrogen peroxide efflux from cells is challenging due to the unknown volume of spaces underneath the cells caused by cellular membrane ruffling.
1:Experimental Design and Method Selection:
The study involved the synthesis of TiO2 nanoparticles through a hydrothermal growth method, followed by characterization using XRD, SEM, and AFM. The ECL of single TiO2 nanoparticles was analyzed in a physiological PBS solution containing L012 and hydrogen peroxide.
2:Sample Selection and Data Sources:
TiO2 nanoparticles with different phases (anatase, rutile) were synthesized and used for ECL imaging. HeLa cells were cultured and used for sensing local hydrogen peroxide efflux.
3:List of Experimental Equipment and Materials:
TiO2 powder (P25) from Degussa Co., L012 from Wako Chemical USA, Inc., Sylgard 184 from Dow Corning, and HeLa cells from the Institute of Biochemistry and Cell Biology.
4:Experimental Procedures and Operational Workflow:
TiO2 nanoparticles were drop-casted onto ITO slides, and ECL imaging was performed using a water objective and an EM CCD camera. A voltage generator was used to apply voltage to the ITO electrode.
5:Data Analysis Methods:
The luminescence intensity from TiO2 nanoparticles was correlated with hydrogen peroxide concentration, and the stability of ECL was assessed over time.
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