Microfluidic devices, solid supports for reagents and related methods

Patent metadata

Field	Value
Publication number	US-9617589-B2
Application number	US-201314402565-A
Country	US
Kind code	B2
Filing date	Mar 15, 2013
Priority date	May 25, 2012
Publication date	Apr 11, 2017
Grant date	Apr 11, 2017

Abstract

A microfluidic device includes a plurality of reaction wells; and a plurality of solid supports, and each of the solid supports has a reagent attached thereto. The reagent is attached to the solid support via a labile reagent/support bond such that the reagent is configured to be cleaved from the support via a cleaving operation.

First claim

That which is claimed is:

1. A microfluidic device for delivering reagent primers to a reaction well using a solid support wherein the reagent primers are connected to the solid support via a functional group that is configured to participate in labile bonds, the device comprising:
a plurality of reaction wells; and
a plurality of solid supports having the reagent primers attached thereto, wherein the plurality of wells and the plurality of solid supports are sized and configured such that only a single one of the plurality of solid supports is retained in a corresponding single one of the plurality of wells, and the microfluidic device comprises a seal which fluidically isolates each of the plurality of wells from each other;
wherein the plurality of reaction wells comprises a reaction solution comprising a sample and, in solution, a detectable label to detect nucleic acid amplicons, and reagent primer cleaved from a respective one of the plurality of solid supports.

2. The microfluidic device of claim 1 , wherein the reagent primer is configured to be cleaved from a respective one of the plurality of solid supports via a cleaving operation, the cleaving operation comprising a thermal operation, an addition of a chemical, the addition of an enzyme, application of an electric potential, an application of light and/or ionizing radiation to the labile bonds.

3. The microfluidic device of claim 1 , wherein the reagent primer comprises a nucleic acid sequence for a nucleic acid transcription and/or amplification reaction.

4. The microfluidic device of claim 3 , wherein the reaction solution further comprises a DNA probe.

5. The microfluidic device of claim 4 , wherein the labile bonds of the reagent primer and support comprise a streptavidin-biotin bond and/or avidin-biotin bond.

6. The microfluidic device of claim 5 , wherein the reagent primer is configured to be cleaved from a respective one of the plurality of solid supports via a cleaving operation and the cleaving operation comprises a thermal operation.

7. The microfluidic device of claim 6 , wherein the streptavidin-biotin bond and/or avidin-biotin bond is configured to release the reagent from the support when the solid supports are incubated at about 50-99° C.

8. The microfluidic device of claim 1 , wherein the solid supports are microbeads.

9. The microfluidic device of claim 1 , wherein the microbeads are magnetic.

10. The microfluidic device of claim 1 , wherein the solid supports comprise a marker configured to identify a property of the solid support and/or reagent primer.

11. The microfluidic device of claim 10 , wherein the marker comprises a predefined support shape, a predefined support size, a magnetic property and/or optical property of the support.

12. The microfluidic device of claim 10 , wherein the marker comprises a fluorescence marker.

13. The microfluidic device of claim 1 , wherein the plurality of wells comprise a solid support containment region and a reaction region, and the solid support containment region is configured to receive a single one of the plurality of solid supports.

14. The microfluidic device of claim 13 , wherein the solid support containment region has a cross-sectional area that generally corresponds to a size and/or shape of the single one of the plurality of solid supports and the solid support containment region is configured such that reactions occur in the solid support containment region and/or the reaction region.

15. The microfluidic device of claim 13 , wherein the plurality of solid supports comprise a first plurality of solid supports and a second plurality of solid supports having a size and/or shape that is different from the first plurality of solid supports, and the solid support containment regions include at least a first plurality of solid support containment regions configured to preferentially retain the first plurality of solid supports and a second plurality of solid support containment regions configured to preferentially retain the second plurality of solid supports.

16. The microfluidic device of claim 13 , wherein the solid support containment regions have a diameter of about 3.5 to 5.0 μm and a depth of about 4.0 to 6.0 μm.

17. The microfluidic device of claim 16 , wherein the plurality of solid supports have a diameter of about 2.0 to about 3.5 μm.

18. The microfluidic device of claim 13 , wherein the plurality of solid supports have a diameter d, and the solid support containment regions have a cylindrical shape having a diameter of 1.05 d to 1.55 d and a depth of 1.2 d to 1.8 d.

19. The microfluidic device of claim 15 , wherein the reaction region has a cross-sectional area that is larger than the cross-sectional area of the solid support containment region.

20. The microfluidic device of claim 13 , wherein the seal comprises a sealing member or an immiscible fluid to fluidically isolate each of the plurality of wells.

21. The microfluidic device of claim 1 , wherein the microfluidic device comprises a membrane in the wells.

22. The microfluidic device of claim 21 , wherein the membrane comprises an extraction membrane.

23. The microfluidic device of claim 22 , wherein the membrane comprises a monolithic aluminum oxide membrane.

24. The microfluidic device of claim 21 , further comprising a channel connected to the membrane that is configured to provide a fluidic connection to the plurality of wells via the membrane.

25. The microfluidic device of claim 21 , further comprising a pattern of microposts between the membrane and the channel to the support the membrane but allow even fluid flow from the plurality of wells via the membrane.

26. The microfluidic device of claim 1 , further comprising a substrate that includes the plurality of reaction wells, wherein the substrate comprises a pattern of hydrophilic and hydrophobic regions, and the plurality of reaction wells are on one of the hydrophilic regions or hydrophobic regions.

27. The microfluidic device of claim 26 , wherein the substrate comprises a first substrate, the device further comprising a second substrate that is spaced apart from the first substrate by a gap and having a corresponding pattern of alternating hydrophilic and hydrophobic regions that face and are aligned with the pattern of alternating hydrophilic and hydrophobic regions on the first substrate.

28. The microfluidic device of claim 27 , wherein the device is configured such that, when an aqueous fluid is positioned in the gap between the first and second substrates and a non-aqueous fluid is subsequently positioned in the gap between the first and second substrate, the aqueous fluid forms droplets isolated by the non-aqueous fluid between the first and second substrates.

29. The microfluidic device of claim 26 , wherein the hydrophobic and hydrophilic regions comprise a patterned, self-assembled monolayer on top of a patterned metal layer on the first and second substrates.

30. The microfluidic device of claim 26 , wherein an electric potential difference applied to the first and/or second substrate effects an electrochemical reaction and/or generates a chemical species through an electrochemical reaction in the plurality of reaction wells, including but not limited to the cleavage of reagents from the solid supports.

31. The microfluidic device of claim 26 ,

Assignees

• Univ North Carolina Chapel Hill

Inventors

• Ramsey John Michael
• Henley William
• Oblath Emily

Classifications

• C12Q2565/629

  being a microfluidic device · CPC title

• B01J2219/00454

  by chemical cleavage from the solid support · CPC title

• B01L2300/0829

  Multi-well plates; Microtitration plates · CPC title

• B01L3/5027

  by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip · CPC title

• C12Q1/686 Primary

  Polymerase chain reaction [PCR] · CPC title