Optochemical fluorescence sensor and method for measuring the concentration of at least one analyte in a sample
First Claim
1. A method for measuring the concentration of at least one analyte in a sample, comprising the steps of(a) contacting the sample with a biorecognitive sensor layer, which is applied on or in close vicinity of at least one island layer consisting of islands of electrically-conductive material,(b) contacting the sample with an analyte-specific fluorescent compound of low quantum yield,(c) binding the analyte-specific fluorescent compound to the analyte, which in turn is bound by the biorecognitive layer, the quantum yield of the analyte-specific fluorescent compound increasing strongly in the vicinity of the island layer,(d) radiating excitation radiation into the at least one island layer, which is suitable for excitation of the analyte-specific fluorescent compound,(e) determining the fluorescence radiation emitted by the bound analyte-specific fluorescent compound as a measure for the analyte concentration.
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Abstract
An optochemical fluorescence sensor with a biorecognitive layer for measuring the concentration of one or more analytes in a sample is provided with at least one island layer which is applied on a sensor substrate. The islands of the island layer are in the form of electrically-conductive material and have a diameter of less than 300 nm, the biorecognitive layer being directly applied on the island layer or bound via a spacer film. In addition, an analyte-specific fluorescent compound is provided which may be added to the sample or is provided in the sensor itself. The biorecognitive layer can bind the analyte to be measured directly or by means of analyte-binding molecules, the originally low quantum yield of the fluorescent compound increasing strongly in the vicinity of the island layer.
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Citations
18 Claims
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1. A method for measuring the concentration of at least one analyte in a sample, comprising the steps of
(a) contacting the sample with a biorecognitive sensor layer, which is applied on or in close vicinity of at least one island layer consisting of islands of electrically-conductive material, (b) contacting the sample with an analyte-specific fluorescent compound of low quantum yield, (c) binding the analyte-specific fluorescent compound to the analyte, which in turn is bound by the biorecognitive layer, the quantum yield of the analyte-specific fluorescent compound increasing strongly in the vicinity of the island layer, (d) radiating excitation radiation into the at least one island layer, which is suitable for excitation of the analyte-specific fluorescent compound, (e) determining the fluorescence radiation emitted by the bound analyte-specific fluorescent compound as a measure for the analyte concentration.
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3. A combination of an optochemical fluorescence measuring apparatus and an analyte-specific fluorescent compound for measuring the concentration of an analyte in a sample,
said optochemical fluorescence measuring sensor comprising a substrate, an island layer consisting of islands of electrically-conductive material on the substrate, said islands having a diameter of less than 300 nm; - a biorecognitive layer applied to said island layer;
said biorecognitive layer being capable of binding with said analyte; andsaid analyte-specific fluorescent compound having a quantum yield which increases strongly in the vicinity of said island layer. - View Dependent Claims (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
- a biorecognitive layer applied to said island layer;
Specification