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

• US 8,883,424 B2
• Filed: 06/07/2013
• Issued: 11/11/2014
• Est. Priority Date: 05/21/1999
• Status: Expired due to Fees

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Chee, Mark S., Dickinson, Todd A., Gunderson, Kevin, O'Neil, Don
• Primary Examiner(s): Chunduru, Prabha
• Application Number:
  US13/912,668
• Publication Number:
  US 20130345067A1
• Time in Patent Office: 522 Days
• Field of Search: None
• US Class Current: 435/6.12
• 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 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• 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 method of sequencing a nucleic acid comprising:

providing a fluidics device comprising;

a first channel in fluid communication with an inlet,

a detection module in fluid communication with said first channel, said detection module comprising an array of wells in communication with a tantalum-containing substrate, a population of microspheres comprising at least a first and a second subpopulation, wherein each subpopulation comprises a bioactive agent and wherein said microspheres are distributed in said wells such that not more than one microsphere is present in each of the wells, and

a second channel in fluid communication with said detection module and an outlet;

providing a fluid comprising a target nucleic acid to the inlet; and

sequencing the target nucleic acid.

2. The method of claim 1, wherein said sequencing comprises detecting an optical signal.

3. The method of claim 1, wherein said sequencing comprises detecting a non-optical signal.

4. The method of claim 1, wherein the first subpopulation and the second subpopulation of microspheres are different sized microspheres.

5. The method of claim 1, wherein the wells are randomly distributed on the tantalum-containing substrate.

6. The method of claim 1, wherein the wells are patterned on the tantalum-containing substrate.

7. The method of claim 1, wherein the bioactive agent comprises a nucleic acid.

8. The method of claim 7, wherein the nucleic acid comprises a primer.

9. The method of claim 8, further comprising an enzyme for extension of the primer.

10. The method of claim 1, wherein the first subpopulation and the second subpopulation of microspheres are different sized microspheres.

11. The method of claim 1, wherein the tantalum-containing substrate comprises a plurality of flow channels.

12. The method of claim 11, wherein at least two flow channels are parallel to each other.

13. The method of claim 1, wherein the tantalum-containing substrate is configured to hold multiple samples.

14. The method claim 1, wherein the wells are randomly distributed on the tantalum-containing substrate.

15. The method of claim 1, wherein the wells are patterned on the tantalum-containing substrate.