Method for manipulating a plurality of plugs and performing reactions therein in microfluidic systems

• US 7,655,470 B2
• Filed: 03/16/2005
• Issued: 02/02/2010
• Est. Priority Date: 10/29/2004
• Status: Active Grant

• Current Assignee: University of Chicago The (University of Chicago)
• Original Assignee: University of Chicago
• Inventors: Gerdts, Cory J., Zheng, Bo, Ismagilov, Rustem F.
• Primary Examiner(s): Gakh; Yelena G
• Application Number:
  US11/082,187
• Publication Number:
  US 20060094119A1
• Time in Patent Office: 1,784 Days
• Field of Search: None
• US Class Current: 436/55
• CPC Class Codes: B01F 25/314 wherein additional components are introduced at the circumference of the conduit
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00522 in a multiple parallel arrangement
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• 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 2300/0838 Capillaries
• 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 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0433 vibrational forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/06 Crystallising dishes
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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 3/527 : for a plurality of reagents
• CPC Class Codes: C30B 29/58 : Macromolecular compounds
• CPC Class Codes: C30B 7/00 : Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: G01N 1/38 : Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 2035/1034 : Transferring microquantities of liquid
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G05D 7/0694 : by action on throttling means or flow sources of very small size, e.g. microfluidics microvalves F16K99/0001; microstructural devices per se B81B
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 436/12 : Condition responsive control

Abstract

The present invention provides microfluidic technology enabling rapid and economical manipulation of reactions on the femtoliter to microliter scale.

Claims

1. A method for manipulating a plurality of plugs comprising:

introducing a first linear array of plugs of a first plug fluid into a microchannel within a microfluidic device at a first inlet with a first flow rate, wherein the plugs of the first plug fluid are separated from each other either by a first carrier fluid and/or by a spacer, and wherein the first carrier fluid and the spacer are substantially immiscible with the first plug fluid and with each other;

introducing an aqueous stream into a second inlet adjacent to the first inlet and in fluid communication with the microchannel with a second flow rate such that a multi-component stream forms, the multi-component stream comprising a laminar flow of the first linear array of plugs and the aqueous stream; and

introducing to the multi-component stream a stream of a second carrier fluid through a third inlet downstream of the first and second inlets with a third flow rate, the third inlet in fluid communication with the microchannel, such that a respective plug, or a portion thereof, of the first linear array of plugs combines with a portion of the aqueous stream to form a second linear array of plugs,

wherein the second carrier fluid is substantially immiscible with the first plug fluid and the spacer.

2. The method of claim 1, wherein the first and second flow rates are adjusted such that the volume of the plugs of the second linear array of plugs is smaller than the volume of the plugs of the first linear array of plugs.

3. The method of claim 1, wherein the first flow rate and the second flow rate are varied.

4. The method of claim 1, wherein the aqueous stream comprises a reagent.

5. The method of claim 4, wherein the reagent is a reagent capable of being crystallized.

6. The method of claim 5, wherein the first linear array of plugs comprises reagents for a sparse matrix screen.

7. The method of claim 5, wherein the reagent is a protein.

8. The method of claim 1, wherein the aqueous stream comprises a laminar flow of a plurality of aqueous solutions.

9. The method of claim 1, wherein either of or both of the first and second carriers comprise a fluorinated fluid.

10. The method of claim 1, wherein the first linear array of plugs is disposed in a pre-formed cartridge before being introduced into the microchannel.

11. A method for manipulating a plurality of plugs comprising:

introducing a first linear array of plugs of a first plug fluid into a microchannel within a microfluidic device at a first inlet with a first flow rate, wherein the plugs of the first plug fluid are separated from each other either by a first carrier fluid and/or by a spacer, and wherein the first carrier fluid and the spacer are substantially immiscible with the first plug fluid and with each other; and

introducing a stream of a second carrier fluid into a second inlet in fluid communication with the microchannel with a second flow rate such that at least one plug of the first linear array of plugs is separated by the stream of the second carrier fluid to form a second linear array of plugs, the second linear array of plugs comprising the resultant plugs formed from the at least one plug of the first linear array of plugs and other plugs of the first linear array of plugs,

wherein the second carrier fluid is substantially immiscible with the first plug fluid and the spacer.

12. The method of claim 11, wherein the size of the plugs of the second linear array of plugs is adjusted by the first flow rate relative to the second flow rate.

13. The method of claim 11, further comprising:

introducing an aqueous stream into a third inlet downstream of the first and second inlets with a third flow rate, the third inlet in fluid communication with the microchannel, such that at least one plug in the second linear array of plugs combines with a portion of the aqueous stream to form a third linear array of plugs, the third linear array of plugs comprising the resultant plugs formed from the at least one plug of the second linear array of plugs and other plugs of the second linear array of plugs.

14. The method of claim 13, wherein the aqueous stream comprises at least one reagent, and the concentration of the reagent in the plugs of the third linear array of plugs is adjusted by the second flow rate relative to the third flow rate.

15. The method of claim 14, wherein the reagent is a protein.

16. The method of claim 14, wherein the concentration of the reagent in the plugs of the third linear array of plugs is varied.

17. The method of claim 11, wherein either of or both of the first and second carriers comprise a fluorinated fluid.

18. The method of claim 11, wherein the first linear array of plugs is disposed in a pre-formed cartridge before being introduced into the microchannel.

19. A method for manipulating a plurality of plugs comprising:

introducing a first linear array of plugs of a first plug fluid into a microchannel within a microfluidic device at a first inlet with a first flow rate, wherein the first linear array of plugs comprises a protein, wherein the plugs of the first plug fluid are separated from each other either by a first carrier fluid and/or by a spacer, and wherein the first carrier fluid and the spacer are substantially immiscible with the first plug fluid and with each other;

introducing a stream of a second carrier fluid into a second inlet in fluid communication with the microchannel with a second flow rate such that the respective plug of the first linear array of plugs is separated by the stream of the second carrier fluid to form a second linear array of plugs, wherein the size of the second linear array of plugs is varied by adjusting the first flow rate relative to the second flow rate, and wherein the second carrier fluid is substantially immiscible with the first plug fluid and the spacer; and

introducing an aqueous stream into a third inlet downstream of the first and second inlets with a third flow rate, the third inlet in fluid communication with the microchannel, such that the respective plug in the second linear array of plugs combines with a portion of the aqueous stream to form a third linear array of plugs, wherein the third linear array of plugs contains different concentrations of the protein originally contained in the first linear array of plugs.

20. A method of conducting a reaction within at least one plug, comprising the steps of:

introducing a carrier-fluid into a first microchannel of a device;

simultaneously introducing at least one first array of plugs and one aqueous stream into a first inlet in fluid communication with the first microchannel so that at least one plug of a second array of plugs forms in the carrier-fluid after the the first array of plugs and the aqueous stream contact the carrier-fluid;

wherein the first array of plugs comprises at least one first reagent;

the aqueous stream comprises a second reagent;

each plug-fluid is immiscible with the carrier-fluid; and

wherein at least one plug of the second array of plugs comprises at least both a portion of one plug of the first array of plugs and a portion of the aqueous stream so that the reaction of the reagents substantially occurs in the second array of plugs; and

each plug is substantially surrounded by carrier.