Self-referencing enzyme-based microsensor and method of use
First Claim
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1. A method of determining a flux of an analyte with respect to a source, the method comprising:
- (a) oscillating an electrode including an enzyme between first and second positions proximal to the source, whereby the enzyme contacts the analyte and generates a species detectable by the electrode, and whereby at each of the positions the concentration of the detectable species is dependent on the concentration of the analyte;
(b) applying a voltage to a portion of the electrode so that the detectable species undergoes a chemical change to generate a position-dependent current;
(c) sensing the current at the first and second positions; and
(d) using the sensed currents to determine the flux of the analyte with respect to the source.
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Abstract
A micron-scale self-referencing microsensor is disclosed which uses an enzyme-based electrode to measure the flux of an electrochemically inactive analyte such as glucose. The electrode oscillates between at least two positions. In the presence of the analyte, the enzyme generates a species detectable by the electrode.
506 Citations
16 Claims
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1. A method of determining a flux of an analyte with respect to a source, the method comprising:
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(a) oscillating an electrode including an enzyme between first and second positions proximal to the source, whereby the enzyme contacts the analyte and generates a species detectable by the electrode, and whereby at each of the positions the concentration of the detectable species is dependent on the concentration of the analyte;
(b) applying a voltage to a portion of the electrode so that the detectable species undergoes a chemical change to generate a position-dependent current;
(c) sensing the current at the first and second positions; and
(d) using the sensed currents to determine the flux of the analyte with respect to the source. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of assessing the viability of an embryo by determining a flux of an analyte with respect to the embryo, the method comprising:
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(a) oscillating an electrode including an enzyme between first and second positions proximal to the embryo, whereby the enzyme contacts the analyte and generates a species detectable by the electrode, and whereby at each of the positions the concentration of the detectable species is dependent on the concentration of the analyte;
(b) applying a voltage to a portion of the electrode so that the detectable species undergoes a chemical change to generate a position-dependent current;
(c) sensing the current at the first and second positions;
(d) using the sensed currents to determine the flux of the analyte with respect to the embryo; and
(e) using the flux determination to assess the viability of the embryo.
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16. A method of screening for compounds that affect a transmembrane flux of an analyte with respect to a cell, the method comprising:
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(a) oscillating an electrode including an enzyme between first and second positions proximal to the cell, whereby the enzyme contacts the analyte and generates a species detectable by the electrode, and whereby at each of the positions the concentration of the detectable species is dependent on the concentration of the analyte;
(b) applying a voltage to a portion of the electrode so that the detectable species undergoes a chemical change to generate a position-dependent current;
(c) sensing the current at the first and second positions;
(d) using the sensed currents to determine the transmembrane flux of the analyte with respect to the cell;
(e) contacting the cell with a test compound;
(f) repeating steps (a) through (d) for the cell in the presence of the test compound; and
(g) determining whether a difference exists between the transmembrane flux of the analyte with respect to the cell in the presence of the test compound and the transmembrane flux of the analyte with respect to the cell in the absence of the test compound, a difference indicating that the test compound affects the transmembrane flux of the analyte with respect to the cell.
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Specification