Microfluidic devices, systems and methods for performing integrated reactions and separations

• US 6,551,836 B1
• Filed: 11/15/1999
• Issued: 04/22/2003
• Est. Priority Date: 06/08/1998
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Chow, Andrea W., Parce, J. Wallace, Kopf-Sill, Anne R., Sundberg, Steven A.
• Primary Examiner(s): LUDLOW, JAN M
• Application Number:
  US09/442,073
• Time in Patent Office: 1,254 Days
• Field of Search: 436/177, 436/180, 436/52, 436/149, 436/161, 436/172, 204/451, 204/453, 435/7.1, 435/7.2, 435/7.4
• US Class Current: 436/149
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: G01N 2001/4038 electric methods, e.g. electromigration, electrophoresis, ionisation
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10T 436/117497 with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

Microfluidic devices for performing integrated reaction and separation operations. The devices have a planar substrate having a first surface with an integrated channel network disposed therein. The reaction region in the integrated microscale channel network has a mixture of at least first and second reactants located therein, wherein the mixture interacts to produce one or more products. The reaction region is configured to maintain contact between the first and second reactants contained within it. The device also includes a separation region in the integrated channel network, where the separation region is configured to separate the first reactant from the product, when the first reactant and product are flowing through the separation region.

Claims

1. A method of analyzing an effect of a first analyte on a second analyte, comprising:

contacting a first analyte with a second analyte in continuous flow mode in a first microfluidic channel portion having substantially no electric field applied across its length, wherein the first analyte and the second analyte interact to produce a product;

transporting at least a portion of the first analyte, second analyte and product to a second channel portion that is in fluid communication with the first channel portion and which has an electric field applied across its length;

detecting a difference in the electrokinetic mobility of the product relative to at least one of the first and second analytes, the difference in mobility being indicative of an effect of the first analyte on the second analyte.

2. The method of claim 1, wherein the effect of the first analyte on the second analyte is a binding of the first analyte to the second analyte, thereby producing a product with a different electrokinetic mobility relative to the first analyte or the second analyte.

3. The method of claim 1, wherein the effect of the first analyte on the second analyte is a cleavage effect, thereby producing a product with a different electrokinetic mobility relative to the first analyte or the second analyte.

4. The method of claim 1, wherein the product comprises a detectable label.

5. The method of claim 4, wherein the detectable label comprises an optically detectable label.

6. The method of claim 5, wherein the optically detectable label comprises a fluorescent label.

7. The method of claim 1, wherein the first analyte, the second analyte and the product are transported into the second microfluidic channel portion by applying a pressure differential between the first channel portion and the second channel portion.

8. The method of claim 1, wherein the difference in electrokinetic mobility results from a change in charge of the product relative to the first or second analytes.

9. The method of claim 1, wherein the first analyte is transported from a first reservoir into the first channel portion, and the second analyte is transported from a second reservoir into the first channel portion, thereby contacting the first analyte with the second analyte in the first microfluidic channel portion.

10. A method of detecting the effect of a third analyte on the interaction between a first and a second analyte, the method comprising:

contacting a first analyte and a second analyte with at least one third analyte in a first microfluidic channel portion having substantially no electric field applied across its length, wherein the first analyte and the second analyte interact to produce a product;

transporting at least a portion of the first analyte, second analyte and product to a second channel portion that is in fluid communication with the first channel portion and which has an electric field applied across its length;

detecting a difference in the electrokinetic mobility of the product relative to at least one of the first and second analytes, the difference in mobility being indicative of an effect of the first analyte on the second analyte; and

,

measuring a change in the electrokinetic mobility of the product in the presence of the at least one third analyte relative to a change in the electrokinetic mobility of the product in the absence of the at least one third analyte.

11. The method of claim 10, wherein the first analyte is transported from a first reservoir into the first channel portion, and the second analyte is transported from a second reservoir into the first channel portion, thereby contacting the first analyte with the second analyte in the a first microfluidic channel portion.

12. The method of claim 10, comprising contacting the first, second and third analytes in a high-throughput screening format.

13. The method of claim 10, comprising contacting the first, second and third analytes in continuous flow mode.

14. The method of claim 10, comprising contacting the first and second analytes and periodically introducing a plurality of third analytes into a continuously flowing stream comprising the first and second analytes.

15. The method of claim 10, wherein the at least one third analyte comprises a test compound.

16. The method of claim 10, wherein the at least one third analyte comprises an affector of a reaction.

17. The method of claim 16, wherein the affector is an inhibitor or an enhancer.

18. The method of claim 10, wherein the first and second analytes are selected from the group comprising:

an enzyme and a substrate, a ligand and a receptor, and a cell affector and a cell.

19. The method of claim 10, wherein the product comprises a detectable label.

20. The method of claim 19, wherein the detectable label comprises an optically detectable label.

21. The method of claim 20, wherein the optically detectable label comprises a fluorescent label.

22. The method of claim 10, wherein the first analyte, the second analyte and the product are transported into the second microfluidic channel portion by applying a pressure differential between the first channel portion and the second channel portion.