Fluorescent reporter beads for fluid analysis
First Claim
1. A method of measuring the concentration of a first analyte in the fluid bulk of a sample fluid, comprising the steps of:
- mixing a plurality of a first type of reporter beads with said sample fluid, each said reporter bead comprising;
a substrate bead; and
a plurality of a first type of reporter molecule immobilized on or in said substrate bead;
wherein an optical property of said reporter molecules is a function of the concentration of said first analyte;
flowing said sample fluid through the measurement zone of a flow cytometer such that said reporter beads flow in substantially a single file;
measuring said optical property of said reporter molecules; and
obtaining the concentration of said first analyte from said optical property.
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Accused Products
Abstract
The present invention provides a method and apparatus for rapid measurement of a fluid bulk analyte, requiring only microscale volumes. Several fluid bulk analytes can be measured simultaneously and, for biological samples, the cell content can also be measured simultaneously. The invention comprises reporter beads for chemical analysis of fluid bulk properties such as pH, oxygen saturation and ion content. Each reporter bead comprises a substrate bead having a plurality of at least one type of fluorescent reporter molecules immobilized thereon. The fluorescent properties of the reporter bead are sensitive to a corresponding analyte. Reporter beads are added to a fluid sample and the analyte concentration is determined by measuring fluorescence of individual beads, for example in a flow cytometer. Alternatively, reporter molecules which change absorbance as a function of analyte concentration can be employed.
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Citations
47 Claims
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1. A method of measuring the concentration of a first analyte in the fluid bulk of a sample fluid, comprising the steps of:
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mixing a plurality of a first type of reporter beads with said sample fluid, each said reporter bead comprising; a substrate bead; and a plurality of a first type of reporter molecule immobilized on or in said substrate bead; wherein an optical property of said reporter molecules is a function of the concentration of said first analyte; flowing said sample fluid through the measurement zone of a flow cytometer such that said reporter beads flow in substantially a single file; measuring said optical property of said reporter molecules; and obtaining the concentration of said first analyte from said optical property. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. A reporter bead cartridge for adding reporter beads to a sample fluid as said fluid is inlet into a flow cytometer, said reporter beads for detection of an analyte in the fluid bulk of said sample fluid, said bead cartridge comprising:
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a cartridge adapted for coupling with the fluid inlet of the flow cytometer, whereby reporter beads disposed therein mix with the sample fluid as it flows through the cartridge and into the flow cytometer; and a plurality of reporter beads disposed within said cartridge, each reporter bead comprising; a substrate bead; and a plurality of a first type of reporter molecule immobilized on or in said substrate bead; wherein an optical property of said reporter molecules is a function of the concentration of said analyte in said fluid bulk. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37)
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38. A cross-sensitivity compensating reporter bead for optical detection of a first analyte in a fluid bulk also containing a second analyte, using a reporter molecule which is intentionally sensitive to said first analyte and incidentally sensitive to said second analyte, comprising:
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a substrate bead; a plurality of a first type of reporter molecules immobilized on or in said substrate bead, wherein an optical property of said first reporter molecules is a function of the concentrations of both said first analyte and said second analyte; and a plurality of a second type of reporter molecule immobilized on or in said substrate beads, wherein an optical property of said second reporter molecules is a function of the concentration of said second analyte. - View Dependent Claims (39, 40, 47)
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41. A cross-sensitivity compensating reporter bead mixture for optical measurement of the concentration of a first analyte in a fluid bulk also containing a second analyte, using a reporter molecule which is intentionally sensitive to said first analyte and incidentally sensitive to said second analyte, said mixture comprising:
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a plurality of a first type of reporter bead, each said bead comprising; a first type of substrate bead; and a plurality of a first type of reporter molecule immobilized on or in said substrate bead, wherein an optical property of said first type of reporter molecules is a function of the concentrations of both said first analyte and said second analyte; and a plurality of a second type of reporter bead, each said bead comprising; a second type of substrate bead; and a plurality of a second type of reporter molecule immobilized on or in said substrate bead, wherein an optical property of said second reporter molecules is a function of the concentration of said second analyte - View Dependent Claims (42, 43, 44, 45, 46)
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Specification