Integrated microfluidic devices

• US 6,007,690 A
• Filed: 07/30/1997
• Issued: 12/28/1999
• Est. Priority Date: 07/30/1996
• Status: Expired due to Term

• Current Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Aclara Biosciences, Inc (Laboratory Corporation Of America Holdings)
• Inventors: Nelson, Robert J., Hooper, Herbert H., Hauser, Alan K., Sassi, Alexander P.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/902,855
• Time in Patent Office: 881 Days
• Field of Search: 204/450, 204/451, 204/452, 204/453, 204/454, 204/455, 204/600, 204/601, 204/602, 204/603, 204/604, 204/605
• US Class Current: 204/601
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027

Abstract

Integrated microfluidic devices comprising at least an enrichment channel and a main electrophoretic flowpath are provided. In the subject integrated devices, the enrichment channel and the main electrophoretic flowpath are positioned so that waste fluid flows away from said main electrophoretic flowpath through a discharge outlet. The subject devices find use in a variety of electrophoretic applications, including clinical assays, high throughput screening for genomics and pharmaceutical applications, point-or-care in vitro diagnostics, molecular genetic analysis and nucleic acid diagnostics, cell separations, and bioresearch generally.

Claims

1. An integrated microfluidic device comprising microchannels formed in a generally planar substrate, said device comprising:

an enrichment microchannel comprising an enrichment channel portion containing an enrichment medium, said enrichment channel portion having a sample inlet, a discharge outlet, an elution inlet, and an enriched fraction fluid outlet;

a main electrophoretic microchannel enclosing a main electrophoretic flowpath, said main electrophoretic microchannel being connected in fluid-conducting relationship to said enriched fraction fluid outlet, and said main electrophoretic microchannel comprising a detection region; and

means for moving a sample into said enrichment channel portion by way of said sample inlet and out from said enrichment channel portion by way of said discharge outlet, whereby at least a fraction of a component of interest in the sample is retained by said enrichment medium;

means for moving at least a portion of the fraction of the component of interest out from said enrichment channel portion by way of said enriched fraction outlet and into said main electrophoretic microchannel, wherein said means for moving the fraction of the component of interest out from said enrichment channel portion and into said main electrophoretic microchannel comprises a first electrode located in said enrichment channel at a point upstream from said elution inlet and a second electrode located downstream from said detection region.

2. The device according to claim 1, further comprising a secondary microchannel connected in fluid-conducting relation between said enrichment fraction fluid outlet and said main electrophoretic microchannel.

3. The device according to claim 2 wherein said secondary microchannel encloses a secondary electrophoretic flowpath.

4. The device according to claim 2 wherein said secondary microchannel and said main electrophoretic microchannel intersect to form an injection cross.

5. An integrated microfluidic device comprising microchannels formed in a generally planar substrate, said device comprising:

an enrichment microchannel comprising an enrichment channel portion containing an enrichment medium, said enrichment channel portion having a first inlet and a first fluid outlet;

a main electrophoretic microchannel enclosing a main electrophoretic flowpath, said main electrophoretic microchannel being connected in fluid-conducting relation to said first fluid outlet, and said main electrophoretic microchannel comprising a detection region;

means for moving a sample into said enrichment channel portion by way of said first inlet and out from said enrichment channel portion by way of said first fluid outlet, whereby at least a fraction of a component of interest in the sample is retained by said enrichment medium;

a discharge channel connected in fluid communication with said first fluid outlet at a point upstream from said main electrophoretic flowpath, for discharging a portion of said sample other than said fraction of the component of interest from said main electrophoretic flowpath; and

means for moving at least a portion of the fraction of the component of interest from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel, wherein said means for moving at least a portion of the fraction of the component of interest out from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel comprises a first electrode located in said enrichment channel upstream of said first inlet and a second electrode located downstream from said detection region.

6. The device according to claim 5, further comprising a secondary microchannel connected in fluid conducting relation between said first fluid outlet and said main electrophoretic microchannel.

7. The device according to claim 6 wherein said secondary microchannel and said main electrophoretic microchannel intersect to form an injection cross.

8. An integrated microfluidic device comprising microchannels formed in a generally planar substrate, said device comprising:

an enrichment microchannel comprising an enrichment channel portion containing an enrichment medium, said enrichment channel portion having a first inlet a first fluid outlet;

a main electrophoretic microchannel enclosing a main electrophoretic flowpath, said main electrophoretic microchannel being connected in fluid-conducting relation to said first outlet, and said main electrophoretic microchannel comprising a detection region;

means for moving a sample into said enrichment channel portion by way of said first inlet and out from said enrichment channel portion by way of said first fluid outlet, whereby at least a fraction of a component of interest in the sample is retained by the enrichment medium;

a discharge channel connected in fluid communication with said first fluid outlet at a point upstream from said main electrophoretic flowpath, for discharging a portion of said sample other than said fraction of the component of interest away from said main electrophoretic flowpath;

means for moving at least a portion of the fraction of the component of interest out from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel, wherein said means for moving at least a portion of the fraction of the component of interest out from said enrichment channel portion by way of said first fluid outlet and into said main electrophoretic microchannel comprises a first electrode located upstream from said first inlet and a second electrode located downstream from said detection region; and

a plurality of input microchannels connected in fluid relation to said first fluid inlet, said plurality of input microchannels comprising at least a sample input microchannel an an elution input microchannel.

9. A microfluidic device for use with a detector to analyze a sample having an analyte portion and a waste portion, the microfluidic device comprising a substrate having a surface and comprising at least one microchannel, an inlet in fluid communication with said microchannel proximal to a first end of said microchannel, and a waste outlet, enrichment means in said microchannel for separating at least a part of the analyte portion from the waste portion, said microchannel having a detection region downstream of the enrichment means accessible by the detector for analyzing a characteristic of the sample, said waste outlet in fluid communication with said microchannel upstream of the detection region and adjacent said enrichment means, transfer means for moving the sample from the inlet to the enrichment means and the waste portion from the enrichment means to the waste outlet and at least first and second spaced-apart electrodes, with one of said electrodes proximal to said inlet and in electrical contact with the sample when the sample is present in the microfluidic device and the other of said electrodes is downstream from said detection region for moving the analyte portion from the enrichment means to the detection means whereby part of the analyte portion but not the waste portion is moved to the detection region for facilitating accurate analysis by the detector of said part of the analyte portion at the detection region.

10. The microfluidic device according to claim 9 wherein the enrichment means includes a chromatographic material.

11. The microfluidic device according to claim 9 wherein the enrichment means includes an electrophoretic gel medium.

12. The microfluidic device according to claim 9 wherein the substrate is provided with an additional inlet in fluid communication with the at least one microchannel upstream of the enrichment means, means for introducing a solution into the additional inlet to contact the enrichment means and separate said part of the analyte portion from the enrichment means.

13. The microfluidic device according to claim 9 wherein the waste outlet is in fluid communication with the at least one microchannel at the enrichment means so that the waste portion exits the at least one microchannel at the enrichment means.

14. The microfluidic device according to claim 9 wherein the at least one microchannel includes a first microchannel portion extending from the inlet to the waste outlet and a second microchannel portion which meets the first microchannel portion at an intersection and extends to the detection region, the enrichment means being at least partially in the intersection.

15. The microfluidic device according to claim 9 wherein the waste outlet is in fluid communication with the at least one microchannel between the enrichment means and the detection region.

16. The microfluidic device according to claim 9 further comprising an electrophoretic medium disposed in the at least one microchannel in the vicinity of the detection region, the waste outlet being in fluid communication with the at least one microchannel between the enrichment means and the electrophoretic medium.

17. The microfluidic device according to claim 9 wherein the transfer means includes a third electrode in electrical contact with the waste portion when the waste portion is in said waste outlet.

18. The microfluidic device according to claim 9 wherein the at least one microchannel includes a first microchannel portion downstream from the enrichment means and a second microchannel portion which meets the first microchannel portion at an intersection and extends to the detection region, the at least first and second spaced-apart electrodes including first and second spaced-apart electrodes for moving said part of the analyte portion from the enrichment means through the first microchannel portion to the intersection and third and fourth spaced-apart electrodes for moving said part of the analyte portion from the intersection through the second microchannel portion to the detection region.

19. The microfluidic device according to claim 9 wherein the substrate is provided with an additional inlet in fluid communication with the at lest one microchannel and wherein the at least one microchannel includes a first microchannel portion extending from the inlet to the enrichment means, a second microchannel portion extending from the additional inlet to intersect the first microchannel portion whereby a fluid other than the sample can be supplied to the at least one microchannel by the additional inlet.

20. The microfluidic device according to claim 9 wherein the substrate is provided with a plurality of additional inlets in fluid communication with the at least one microchannel and wherein the at least one microchannel includes a first microchannel portion extending from the inlet to the enrichment means, a plurality of additional microchannel portions extending from the additional inlets to intersect the first microchannel portion whereby a plurality of fluids other than the sample can be supplied to the at least one microchannel by the additional inlets.

21. The microfluidic device according to claim 9, wherein said microchannel comprises an additional detection region, downstream from said waste outlet.

22. The microfluidic device according to claim 21 wherein the analyte portion includes a plurality of different fractions of interest, the device further comprising an additional enrichment means in said microchannel between said enrichment means and said inlet, said additional enrichment means enhancing a different fraction of interest in the analyte portion.

23. The microfluidic device according to claim 22 wherein the analyte portion includes a plurality of different nucleic acid fragments and wherein the additional enrichment means each includes a molecular probe for capturing one of the different nucleic acid fragments.

24. The microfluidic device according to claim 21, wherein the two detection regions are disposed in series along said microchannel.

25. The microfluidic device according to claim 21, wherein the two detection regions are disposed in parallel along said microchannel.

26. The microfluidic device according to claim 25 wherein said microchannel is connected to a branch microchannel, said branch microchannel comprising two detection regions.

27. The microfluidic device according to claim 25 wherein the at least one microchannel includes a first microchannel portion and a plurality of second microchannel portions which meet the first microchannel portion at respective intersections spaced-apart along the first microchannel portion, one of the first-named and additional detection regions being in each of the plurality of second microchannel portions.

28. The microfluidic device according to claim 27 wherein the at least first and second spaced-apart electrodes includes first and second spaced-apart electrodes for each of the second microchannel portions to move said part of the analyte portion through the respective first-named or additional detection region.

29. A device according to claim 9 for use with a sample having first and second analyte portions wherein said microchannel includes a branched microchannel portion having first and second branch microchannel portions, said inlet being upstream of the branched microchannel portion, second enrichment means being in said first branch microchannel portion for separating at least part of the first analyte portion from the second analyte portion, said first branch microchannel portion comprising a second detection region downstream from said second enrichment means and in fluid communication with a waste outlet downstream of and proximal to said enrichment means, the first and second spaced-apart electrodes positioned for moving said part of the first analyte portion from the enrichment means to the detection region.

30. The microfluidic device according to claim 29 wherein each of the first and second branch microchannel portions includes a first microchannel segment downstream from the respective enrichment means and a second microchannel segment which meets the first microchannel segment at an intersection and extends to the respective detection region, the at least first and second spaced-apart electrodes including first and second spaced-apart electrodes for moving said part of the first analyte portion from the first-named enrichment means through the first microchannel segment to the intersection and at least a first additional electrode for moving said part of the first analyte portion from the intersection through the second segment portion to the first-named detection region and the at least a third electrode including at least a third electrode for moving said part of the second analyte portion from the additional enrichment means through the first microchannel segment to the intersection and at least a second additional electrode for moving said part of the second analyte portion from the intersection through the second segment portion to the additional detection region.

31. The microfluidic device according to claim 9 wherein the enrichment means includes affinity binding capture and release molecules to capture at least a part of the analyte portion so as to separate at least a part of the analyte portion from the waste portion.

32. The microfluidic device according to claim 31 wherein the affinity binding capture and release molecules are bound to a plurality of bodies capable of producing a magnetic field.

33. The microfluidic device according to claim 32 further comprising a magnet for coupling magnetically to at least some of the bodies capable of producing a magnetic field.

34. A microfluidic device for use with first and second detectors to analyze a first analyte portion and a second analyte portion in a sample, said microfluidic device comprising a substrate having a surface and being provided with a microchannel network formed therein, the microchannel network comprising a main microchannel including a branched microchannel portion connected to first and second branch microchannel portions, first enrichment means in the first branch microchannel portion for separating at least part of the first analyte portion from the second analyte portion, second enrichment means in the second branch microchannel portion for separating at least part of the second analyte portion from the first analyte portion, the first branch microchannel portion having a first detection region downstream of the first enrichment means accessible by the first detector for analyzing a characteristic of the first analyte portion, the second microchannel portion having a second detection region downstream of the second enrichment means accessible by the second detector for analyzing a characteristic of the second analyte portion, first and second spaced-apart electrodes in electrical contact with an electrophoretic medium in said microchannel network for moving said part of the first analyte portion from the first enrichment means to the first detection region and at least a third electrode in electrical contact with the electrophoretic medium for moving the second analyte portion from the second enrichment means to the second detection region whereby the first analyte portion can be analyzed by the first detector at the first detection region and the second analyte portion can be analyzed by the second detector at the second detection region.

35. The microfluidic device according to claim 34 wherein the substrate has a first waste outlet in fluid communication with the first branch microchannel portion upstream of the first detection region for removing the second analyte portion from the first branch microchannel portion so as not to contaminate the first detection region with the second analyte portion and wherein the substrate has a second waste outlet in fluid communication with the second branch microchannel portion upstream of the second detection region for removing the first analyte portion from the second branch microchannel portion so as not to contaminate the second detection region with the first analyte portion.

36. The microfluidic device according to claim 35 wherein the substrate is provided with an outlet reservoir in fluid communication with the first and second waste outlets.