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US 2013/0157378 A1Published application
Attenuating dye for interrogating multiple surfaces, and method thereof
OwnerJames Lawrence Burginventor
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Abstract
A device including a shallow chamber for analyzing a plurality of target analytes in a body fluid using the signal generated by fluorescent detector molecules each specific for a target analyte, an attenuating dye for attenuating the signal emitted by fluorescent detector molecules specifically bound to the surfaces of the chamber other than an optically clear surface, and method for determining the signal generated by each of the plurality of analytes.
Claims27
A method for attenuating non-specific fluorescence in a microfluidic device for detecting the presence of multiple target analytes, comprising:
- (i) providing a microfluidic device having an assay chamber comprising a first wall, wherein at least a portion of said first wall is optically clear, a wall opposite to the optically clear wall, and a lumen, the luminal surface of said optically clear wall coated with first binding partners specific for a first target analyte in a biological specimen and the luminal surface of said opposite wall coated with second binding partners specific for a second target analyte;
- (ii) introducing into said chamber a third binding partner for said first target analyte, said third binding partner labeled with a fluorophore detector molecule, and a fourth binding partner for said second target analyte said fourth binding partner labeled with said fluorophore detector molecule;
- (iii) incubating to allow binding events to occur;
- (iv) removing the contents of said chamber;
- (v) optically measuring fluorescence in said chamber, wherein the optical measurement is related to the concentration of the first and second target analytes;
- (vi) introducing a solution comprising a dye into the chamber, wherein the dye absorbs light of a wavelength range selected from the group consisting of emission wavelength range, excitation wavelength range, and their combination of said fluorescent detector molecule
- (vii) optically measuring fluorescence in said chamber, wherein the optical measurement is related to the concentration of the first target analyte;
- (viii) calculating the concentration of the second target analyte from the optical measurements of step (v) and step (vii).
5 more elements
A composition of matter, comprising:
- a microfluidic device comprising an assay chamber for detecting a target analyte, said assay chamber comprising a first wall wherein at least a portion of said first wall is optically clear, a will opposite to the optically clear wall, and a lumen, the luminal surface of said optically clear wall coated with first binding partners specific for a first target analyte in a sample and the luminal surface of said opposite wall coated with second binding partners specific for a second target analyte in the sample;
- a third binding partner for said first target analyte labeled with a fluorophore detector molecule, and a fourth binding partner for said second target analyte labeled with said fluorophore detector molecule;
- a solution comprising a dye, the dye capable of absorbing light of a wavelength range selected from the group consisting of emission wavelength range, excitation wavelength range, or their combination of any said fluorescent detector label that is hound to the luminal surface of said chamber.
A method for detecting the presence of multiple target analytes in an assay chamber, comprising:
- (i) providing a microfluidic device having an assay chamber comprising a first wall, wherein at least a portion of said first wall is optically clear, a wall opposite to the optically clear wall, and a lumen, the luminal surface of said optically clear wall coated with first binding partners specific for a first target analyte in a biological specimen and the luminal surface of said opposite wall coated with second binding partners specific for a second target analyte in said biological specimen;
- (ii) introducing said biological specimen into the chamber
- (iii) introducing into said chamber a third binding partner for said first target analyte, said third binding partner labeled with a fluorophore detector molecule, and a fourth binding partner for said second target analyte said fourth binding partner labeled with said fluorophore detector molecule;
- (iv) incubating to allow binding events to occur;
- (v) removing the contents of said chamber;
- (vi) optically measuring fluorescence in said chamber, wherein the optical measurement is related to the concentration of the first and second target analytes;
- (vii) introducing a solution comprising a dye into the chamber, wherein the dye absorbs light of a wavelength range selected from the group consisting of emission wavelength range, excitation wavelength range, and their combination of said fluorescent detector molecule
- (viii) optically measuring fluorescence in said chamber, wherein the optical measurement is related to the concentration of the first target analyte;
- (ix) calculating the concentration of the second target analyte from the optical measurements of step (vi) and step (viii).
6 more elements
Dependencies are inferred from the claim text.