Use of microfluidic systems in the detection of target analytes using microsphere arrays

• US 20070184456A1
• Filed: 07/23/2004
• Published: 08/09/2007
• Est. Priority Date: 05/21/1999
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Gunderson, Kevin, Dickinson, Todd, Chee, Mark, Stuelpnagel, John, O'Neil, Don
• Granted Patent: US 8,080,380 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

The invention relates generally to methods and apparatus for conducting analyses, particularly microfluidic devices for the detection of target analytes.

Claims

1. A microfluidic device for the detection of target analytes comprising:

a) a sample inlet port;

b) a detection inlet port;

c) a substrate comprising;

i) at least one sample handling well;

ii) a first microchannel to allow fluid contact between said sample inlet port and said sample handling well;

iii) a detection module comprising;

(a) a plurality of discrete sites in said substrate; and

(b) a population of microspheres comprising at least a first and a second subpopulation, wherein each subpopulation comprises a bioactive agent;

 

wherein said microspheres are distributed on said discrete sites, wherein each discrete site contains not more than one microsphere; and

iv) a second microchannel to allow fluid contact between said sample handling well and said detection inlet port.

3. The device according to claim 1 or claim 2, wherein said substrate comprises a fiber optic bundle.

4. The device according to claim 1 or claim 2, wherein each subpopulation further comprises an optical signature.

5. The device according to claim 1 or claim 2, wherein each subpopulation further comprises an identifier binding ligand that will bind a decoder binding ligand such that the identification of the bioactive agent can be elucidated.

6. The device according to claim 1 or claim 2, wherein said bioactive agent is a nucleic acid.

7. The device according to claim 1 or claim 2, wherein said substrate comprises vertical microstructures between said discrete sites.

8. The device according to claim 1 or claim 2, further comprising a device for regulating sample flow.

9. The device according to claim 8, wherein said device comprises a closed looped channel.

10. The device according to claim 1 or claim 2, wherein each subpopulation further comprises a decoder binding ligand bound to said bioactive agent.

2. A microfluidic device comprising a substrate comprising:

a) a plurality of sample handling wells;

b) a detection channel comprising;

i) a plurality of discrete sites; and

ii) a population of microspheres comprising at least a first and a second subpopulation, wherein each subpopulation comprises a bioactive agent, wherein said microspheres are distributed on said discrete sites; and

c) at least one microchannel to allow fluid contact between each of said sample handling wells and said detection channel.

11. A method of assembling a detector in a microfluidic device comprising:

a) providing a microfluidic device comprising;

i) a first microchannel to allow fluid contact between a sample inlet port and a sample handling well;

ii) a second microchannel to allow fluid contact between said sample handling well and a detection inlet port; and

ii) a detection module comprising a substrate with a surface comprising discrete sites;

b) flowing a fluid across said substrate, said fluid comprising a population of microspheres comprising at least a first and a second subpopulation, wherein each subpopulation comprises a bioactive agent, whereby said beads flow across said discrete sites, and are deposited randomly in said discrete sites; and

c) reversing the flow of said fluid.

12. The method according to claim 11, wherein said microfluidic device further comprises a pump to pump said fluid.

13. The method according to claim 12, wherein said pump is selected from the group consisting of an electrohydrodynamic pump, electrokinetic pump and electroosmotic pump.

14. A method of assembling a detector in a microfluidic device comprising:

a) providing a microfluidic device comprising;

i) a plurality of first micro channels;

ii) a population of microspheres in each of said microchannels; and

iii) at least one receiving chamber connected to said microchannels.

b) flowing said microspheres through said microchannels into said receiving chamber.

15. The method according to claim 14, wherein said receiving chamber is a detection module.

16. The method according to claim 15, wherein said detection module comprises a substrate with a surface comprising discrete sites.

17. The method according to claim 16, wherein said population of microspheres is randomly distributed on said substrate.

18. The method according to claim 14, wherein said population comprises a first and second subpopulation whereby said first and second subpopulations of microspheres are in said first and second microchannels.