Method for obtaining a collection of plugs comprising biological molecules

• US 8,273,573 B2
• Filed: 02/09/2011
• Issued: 09/25/2012
• Est. Priority Date: 05/09/2002
• Status: Expired due to Term

• Current Assignee: University of Chicago
• Original Assignee: University of Chicago
• Inventors: Tice, Joshua David, Gerdts, Cory John, Zheng, Bo, Ismagilov, Rustem F
• Primary Examiner(s): Gakh, Yelena G
• Application Number:
  US13/024,171
• Publication Number:
  US 20110174622A1
• Time in Patent Office: 594 Days
• Field of Search: 436164-166, 436/172, 435/6.12, 435/6.19
• US Class Current: 436/52
• CPC Class Codes: B01D 2009/0086 Processes or apparatus therefor
• CPC Class Codes: B01D 9/0072 Crystallisation in microfluidic devices
• CPC Class Codes: B01F 25/433 Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• 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 14/00 Chemical processes in general for reacting liquids with liquids; Apparatus specially adapted therefor B01J8/00, B01J19/08 take precedence
• 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 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 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00736 Non-biologic macromolecules, e.g. polymeric compounds
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00837 comprising coatings other than catalytically active coatings
• CPC Class Codes: B01J 2219/0086 Dimensions of the flow channels
• CPC Class Codes: B01J 2219/00869 Microreactors placed in parallel, on the same or on different supports
• CPC Class Codes: B01J 2219/00889 : Mixing micromixers B01F33/30
• CPC Class Codes: B01J 2219/00891 : Feeding or evacuation
• CPC Class Codes: B01J 2219/00894 : More than two inlets
• CPC Class Codes: B01J 2219/00903 : Segmented flow
• CPC Class Codes: B01J 2219/00975 : Ultraviolet light
• CPC Class Codes: B01J 2219/00977 : Infrared light
• CPC Class Codes: B01L 2200/0673 : Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/12 : Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0867 : Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 3/502784 : 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: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07K 14/43 : Sweetening agents, e.g. thaumatin, monellin
• CPC Class Codes: C12N 9/2462 : Lysozyme (3.2.1.17)
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Y 302/01017 : Lysozyme (3.2.1.17)
• CPC Class Codes: C30B 29/54 : Organic compounds
• CPC Class Codes: C30B 29/58 : Macromolecular compounds
• CPC Class Codes: C30B 7/14 : the crystallising materials being formed by chemical reactions in the solution
• CPC Class Codes: G01N 1/28 : Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• 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: Y10T 117/10 : Apparatus
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

The present invention provides microfabricated substrates and methods of conducting reactions within these substrates. The reactions occur in plugs transported in the flow of a carrier-fluid.

Claims

1. A method for obtaining a collection of plugs comprising at least one biological molecule and, optionally, at least one or more chemical reactants for conducting a reaction with the biological molecule, the method comprising the steps of:

providing a microfluidic system comprising at least a first and second channel, the first and second channels configured to intersect with each other at a junction, and an outlet adapted for receiving a segment of tubing or a sample tube;

continuously flowing an aqueous fluid containing the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule, through the first channel;

continuously flowing a carrier fluid immiscible with the aqueous fluid through the second channel;

forming a plurality of the plugs of the aqueous fluid containing the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule, by partitioning the aqueous fluid with the flowing immiscible carrier fluid at the junction of the first and second channels, the plugs being substantially surrounded by the immiscible carrier fluid flowing through the channel, wherein at least one plug of the plurality comprises the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule; and

collecting the plurality of the plugs in the segment of tubing or the sample tube, wherein the plugs remain separated by the carrier immiscible fluid.

2. The method according to claim 1, wherein the immiscible carrier fluid is an oil.

3. The method according to claim 2, wherein the oil comprises a surfactant.

4. The method according to claim 3, wherein the surfactant is a fluorosurfactant.

5. The method according to claim 2, wherein the oil is a fluorinated oil.

6. The method according to claim 1, wherein the biological molecule is DNA or RNA.

7. A method for obtaining a collection of plugs comprising at least one biological molecule and, optionally, at least one or more chemical reactants for conducting a reaction with the biological molecule, the method comprising the steps of:

providing a microfluidic device comprising at least a first and second channel, the first and second channels configured to intersect with each other at a junction, and an outlet;

continuously flowing an aqueous fluid containing the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule, through the first channel;

continuously flowing a carrier fluid immiscible with the aqueous fluid through the second channel;

forming a plurality of the plugs of the aqueous fluid containing the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule, by partitioning the aqueous fluid with the flowing carrier immiscible fluid at the junction of the first and second channels, the plugs being substantially surrounded by the immiscible carrier fluid flowing through the channel, wherein at least one plug of the plurality comprises the at least one biological molecule and, optionally, the at least one or more reagents for conducting the reaction with the biological molecule; and

dispensing the plurality of the plugs into a segment of tubing or a sample tube from the outlet using a driving force, wherein the plugs in the segment of tubing or the sample tube remain separated by the carrier immiscible fluid.

8. The method according to claim 7, wherein the immiscible carrier fluid is an oil.

9. The method according to claim 8, wherein the oil comprises a surfactant.

10. The method according to claim 9, wherein the surfactant is a fluorosurfactant.

11. The method according to claim 8, wherein the oil is a fluorinated oil.

12. The method according to claim 7, wherein the driving force is a positive or negative pressure.

13. The method according to claim 7, wherein the biological molecule is DNA or RNA.