Microfluidic detection of analytes
First Claim
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1. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
- providing an aqueous sample in a large volume reservoir comprising a first microelectrode operably attached thereto, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte of interest is charged or has been associated with a charged molecule;
electrophoresing the analyte via a connector from the large volume reservoir to a staging reservoir positioned in a microfluidic device, wherein the connector fluidically connects the large volume reservoir and the staging reservoir, and wherein the microfluidic device comprises an analysis area, a second microelectrode located in the staging reservoir and a third microelectrode located in or distal to at least a portion of the analysis area, said microelectrodes being configured such that an electric field may be generated between the electrodes and a charged analyte electrophoresed from the first microelectrode to the second microelectrode and from the second microelectrode to the third microelectrode; and
transporting the analyte to the analysis area for a time sufficient to result in a higher concentration of analyte in the analysis area than in the sample.
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Abstract
An apparatus and methods for concentrating samples for application to microfluidic devices are disclosed. The methods involve electrophoresing charged molecules from a high volume sample into a smaller volume. The analyte of interest can be a charged molecule or can be modified to be charged using, for example, one or more ionic moieties.
45 Citations
42 Claims
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1. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir comprising a first microelectrode operably attached thereto, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte of interest is charged or has been associated with a charged molecule; electrophoresing the analyte via a connector from the large volume reservoir to a staging reservoir positioned in a microfluidic device, wherein the connector fluidically connects the large volume reservoir and the staging reservoir, and wherein the microfluidic device comprises an analysis area, a second microelectrode located in the staging reservoir and a third microelectrode located in or distal to at least a portion of the analysis area, said microelectrodes being configured such that an electric field may be generated between the electrodes and a charged analyte electrophoresed from the first microelectrode to the second microelectrode and from the second microelectrode to the third microelectrode; and transporting the analyte to the analysis area for a time sufficient to result in a higher concentration of analyte in the analysis area than in the sample. - View Dependent Claims (2, 3, 4, 5)
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6. A method for introducing an analyte of interest to a microfluidic device, comprising:
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introducing a solid phase hound analyte into a large volume reservoir to provide a sample, said sample having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule; exposing the solid phase-bound analyte to conditions sufficient to free the analyte from the solid phase; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analysis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and transporting the analyte to the analysis area for a time sufficient to result in a higher concentration of analyte in the analysis area than in the sample, wherein the analyte is continuously flowed from the large volume reservoir to the analysis area during said time. - View Dependent Claims (7, 8, 9, 10)
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11. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample, wherein the large volume reservoir is a chamber that is not integral with said microfluidic device. - View Dependent Claims (12, 13, 14, 15)
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16. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample, wherein said large volume reservoir is a well of a microwell plate, an eppendorf tube, or a test tube. - View Dependent Claims (17, 18, 19, 20)
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21. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein said analyte is covalently bound to an ionic moiety or is associated with a charged molecule by being bound to a carrier molecule that is ionically charged or is modified to be ionically charged; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample. - View Dependent Claims (22, 23, 24, 25, 26)
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27. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample, wherein the analysis area comprises a capture site comprising a capture agent, and said method further comprises transporting the analyte across the capture site under conditions where the analyte is captured by the capture agent. - View Dependent Claims (28, 29, 30, 31, 32, 33)
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34. A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
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providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule; electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample, wherein the method further comprises immobilizing said analyte of interest on a microparticle before electrophoresis. - View Dependent Claims (35, 36, 37, 38, 39, 40, 41, 42)
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Specification