Microfluidic matrix localization apparatus and methods

• US 6,306,590 B1
• Filed: 06/08/1998
• Issued: 10/23/2001
• Est. Priority Date: 06/08/1998
• Status: Expired due to Term

• Current Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Original Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Inventors: Kopf-Sill, Anne R., Mehta, Tammy Burd
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): PHAM, MINH QUAN K
• Application Number:
  US09/093,832
• Time in Patent Office: 1,233 Days
• Field of Search: 422/50, 422/54, 422/55, 422/56, 422/57, 422/58, 422/68.1, 422/69, 422/82.02, 422/82-5-, 422/129, 422/131, 435/91.2, 435/91.1, 435/6, 435/5, 435/291, 435/810, 435/90-91, 435/91.51, 435/91.5, 435/173.1, 435/174, 435/176-177, 435/283.1, 435/285--28, 435/287--28, 435/287--28, 435/808, 435/814, 436/22.1, 436/23.1, 436/25.1, 436/518, 436/524, 436/525, 436/528, 436/531-532, 436/63, 436/164-6, 436/169, 436/172, 436/175, 436/805
• US Class Current: 435/6.19
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00828 Silicon wafers or plates
• CPC Class Codes: B01J 2219/00831 Glass
• CPC Class Codes: B01J 2219/00833 Plastic
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/00873 Heat exchange
• CPC Class Codes: B01J 2219/00889 Mixing
• CPC Class Codes: B01J 2219/00891 Feeding or evacuation
• CPC Class Codes: B01J 2219/00909 using filters
• CPC Class Codes: B01J 2219/00912 by electrophoresis
• CPC Class Codes: B01J 2219/00961 Temperature
• CPC Class Codes: B01J 2219/00986 Microprocessor
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• 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 2300/1827 using resistive heater
• 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 3/5027 : 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 B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10S 435/814 : Enzyme separation or purification

Abstract

Multiphasic microfluidic apparatus for performing product fluid manipulation and separation in a single continuous unit are provided. Related methods, kits, and compositions are also provided.

Claims

1. A method of performing a polymerase chain reaction in a microfluidic apparatus, said method comprising:

providing a microfluidic device comprising at least two intersecting channels in fluid communication;

filling at least one of said two intersecting channels with an electrophoretic phase selected from the group consisting of a sieving matrix and a molecular partition matrix, wherein the electrophoretic phase comprise at least two PCR reactants and a thermostable polymerase dispersed therein;

performing the polymerase chain reaction in the presence of the electrophoretic phase to produce at least a first product.

2. The method of claim 1, said method further comprising a step of separating the at least first product from the reactants in the presence of the electrophoretic phase.

3. The method of claim 1, wherein at least one of said intersecting channels comprise a reaction region.

4. The method of claim 1, wherein the microfluidic device comprises a high-resistance channel region for heating the at least two PCR reactants.

5. The method of claim 1, wherein the two PCR reactants are heterogeneously dispersed throughout at least a portion of the electrophoretic phase.

6. The method of claim 1, wherein the two PCR reactants are homogeneously dispersed throughout at least a portion of the electrophoretic phase.

7. The method of claim 1, wherein at least one of the two PCR reactants is heterogeneously dispersed in at least a portion of the electrophoretic phase and one of the two PCR reactants is homogeneously dispersed throughout the electrophoretic phase.

8. A method of performing a polymerase chain reaction in a microfluidic apparatus, said method comprising:

providing a microfluidic device comprising at least two intersecting channels in fluid communication;

filling at least one of said two intersecting channels with a mixture comprising a sieving matrix, a thermostable polymerase and a plurality of PCR reaction components;

performing the PCR in the presence of the sieving matrix to produce at least a first product.

9. The method of claim 8, said method further comprising a step of separating the at least first product from the PCR reaction components in the presence of the sieving matrix.

10. The method of claim 8, wherein the step of separating is carried out in a channel region different from a channel region for performing the PCR.

11. The method of claim 8, wherein the sieving matrix is selected from a group consisting of agarose, linear polyacrylamide, methylcellulose, polyethylene oxide and hydroxy ethyl cellulose.

12. The method of claim 8, wherein one of said at least two intersecting channels have an interior dimension of between about 0.1 μ

m and 500 μ

m.

13. The method of claim 8, wherein the two PCR reactants are heterogeneously dispersed throughout at least a portion of the sieving matrix.

14. The method of claim 8, wherein the two PCR reactants are homogeneously dispersed throughout at least a portion of the sieving matrix.

15. The method of claim 8, wherein at least one of the two PCR reactants is heterogeneously dispersed in at least a portion of the sieving matrix and one of the two PCR reactants is homogeneously dispersed throughout the sieving matrix.

16. The method of claim 8, wherein one of said at least two intersecting channels comprise a reaction region.

17. The method of claim 16, wherein the reaction region is substantially filled with the sieving matrix.

18. The method of claim 8, wherein one of said at least two intersecting channels comprise a separation region.

19. The method of claim 18, wherein the separation region is substantially filled with the sieving matrix.