Abstract
In this paper, phosphate assay kit in one cell is realized for the electrochemical detection of intracellular phosphate ions at single cells. The components of the phosphate assay kit, including maltose phosphorylase, maltose, mutarotase, and glucose oxidase, are electrochemically injected into a living cell through a nanometer-sized capillary with the ring electrode at the tip. These components react with phosphate ions inside the cell to generate hydrogen peroxide that is electrochemically oxidized at the ring electrode for the qualification of intracellular phosphate ions. An average 1.7 nA charge was collected from eight individual cells, suggesting an intracellular phosphate concentration of 2.1 mM. The establishment in the electrochemical measurement of phosphate ions provides a special strategy to monitor the fluctuation of intracellular phosphate at single cells, which is significant for the future investigation of phosphate signal transduction pathway.
Highlights
Inorganic phosphate ions are the most abundant anions inside the cells that are essential for nucleic acid and phospholipid biosynthesis, as well as for energy metabolism (Bevington et al, 1992)
When one kit component is removed from the mixed solution inside the capillary, this charge increase disappears. Both of the results suggest the occurrence of chemical reactions of kit components and phosphate ions outside the tip that generates hydrogen peroxide and the following charge increase
The electrochemical detection of intracellular phosphate ions at single cells is realized by the loading of phosphate assay kit into one cell
Summary
Inorganic phosphate ions are the most abundant anions inside the cells that are essential for nucleic acid and phospholipid biosynthesis, as well as for energy metabolism (Bevington et al, 1992). The loading of these kit components into one cell to initialize the reaction with intracellular phosphate and the immediate detection of the product (e.g., hydrogen peroxide) should provide an alternative strategy to detect phosphate ions at single cells. The loading of the phosphate assay kit in one cell is feasible, and the detection of phosphate ions at single cells could be realized.
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