# US 10,118,174 B2 · Device and method for pressure-driven plug transport and reaction

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- Number: US 10,118,174 B2
- Document ID: US10118174B2
- Status: Granted
- Kind: B2
- Application number: 15/825,689
- Filed: 2017-11-29
- Published: 2018-06-07
- Granted: 2018-11-06
- Earliest claimed: 2002-05-09
- Assignees: The University of Chicago
- Applicants: The University of Chicago
- Inventors: Rustem F. Ismagilov; Joshua David Tice; Helen Song Baca; Lewis Spencer Roach
- Web page: https://patentgrep.com/patent/US10118174B2

## CPC

- B01L 3/502784: {specially adapted for droplet or plug flow, e.g. digital microfluidics}
- B01F 23/45: using flow mixing
- 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
- B01F 25/4331: {Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow}
- 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}
- 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)}
- B01J 19/0093: {Microreactors, e.g. miniaturised or microfabricated reactors (laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027)}
- B01L 3/502746: {characterised by the means for controlling flow resistance, e.g. flow controllers, baffles or throttle valves}
- B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- C12P 19/34: Polynucleotides, e.g. nucleic acids, oligoribonucleotides
- C12Q 1/44: involving esterase
- G01N 15/1404: Handling flow, e.g. hydrodynamic focusing
- G01N 35/08: using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
- G01N 35/085: {Flow Injection Analysis}
- B01F 25/31: in conduits or tubes through which the main component flows
- B01F 2101/23: Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
- B01J 2219/00286: Reactor vessels with top and bottom openings
- B01J 2219/00576: fluorophore
- B01J 2219/00585: Parallel processes
- B01J 2219/0059: Sequential processes
- B01J 2219/00599: Solution-phase processes
- B01J 2219/00722: Nucleotides
- B01J 2219/00725: Peptides
- B01J 2219/00736: Non-biologic macromolecules, e.g. polymeric compounds
- B01J 2219/0074: Biological products
- B01J 2219/00756: Compositions, e.g. coatings, crystals, formulations
- B01J 2219/00783: Laminate assemblies, i.e. the reactor comprising a stack of plates
- B01J 2219/00837: comprising coatings other than catalytically active coatings
- B01J 2219/0086: Dimensions of the flow channels
- B01J 2219/00867: Microreactors placed in series, on the same or on different supports
- B01J 2219/00869: Microreactors placed in parallel, on the same or on different supports
- B01J 2219/00889: Mixing
- B01J 2219/00891: Feeding or evacuation
- B01J 2219/00975: Ultraviolet light
- B01J 2219/00977: Infrared light
- B01L 2200/0647: Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L 2200/0668: Trapping microscopic beads
- B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution
- B01L 2300/0883: Serpentine channels
- B01L 2400/0424: Dielectrophoretic forces
- B01L 2400/0448: Marangoni flow; Thermocapillary effect
- B01L 2400/0454: radiation pressure, optical tweezers
- B01L 2400/0487: fluid pressure, pneumatics
- B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites
- G01N 15/1409: Handling samples, e.g. injecting samples
- G01N 15/1484: {microstructural devices}
- Y02A 90/10: Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation
- Y10T 436/118339: with formation of a segmented stream
- Y10T 436/12: Condition responsive control
- Y10T 436/2575: Volumetric liquid transfer
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01F: MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING (mixing paints B44D3/06)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## 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

### Claim 1

A method of conducting an autocatalytic reaction comprising forming a plurality of plugs and conducting the autocatalytic reaction in at least one plug, wherein:
each plug comprises an aqueous fluid surrounded by a hydrophobic fluid;
the aqueous fluid of at least one plug comprises a biological reagent;
the hydrophobic fluid of each plug comprises a fluorosurfactant; and
the fluorosurfactant is linked to an oligoethylene glycol (OEG).

### Claim 2 (depends on 1)

The method of claim 1, wherein the plugs are formed in a microchannel.

### Claim 3 (depends on 1)

The method of claim 1, further comprising sorting the plugs.

### Claim 4 (depends on 1)

The method of claim 1, wherein the autocatalytic reaction is a polymerase-chain reaction (PCR).

### Claim 5 (depends on 1)

The method of claim 1, wherein at least one plug comprises a single biological reagent.

### Claim 6 (depends on 5)

The method of claim 5, wherein the single biological reagent is DNA or RNA.

### Claim 7 (depends on 1)

The method of claim 1, wherein the fluorosurfactant comprises an oligoethylene glycol terminal.

### Claim 8 (depends on 7)

The method of claim 7, wherein the fluorosurfactant is of the formula:
wherein m is 0, 3, 4, 5, or 6; and n is an integer of no less than 16.

### Claim 9 (depends on 7)

The method of claim 7, where the oligoethylene glycol is linked to a terminal end of the fluorosurfactant.

### Claim 10 (depends on 1)

The method of claim 1, wherein the fluorosurfactant is linked to the oligoethylene glycol (OEG).

### Claim 11

A method of preparing a composition comprising a plurality of plugs, the method comprising:
flowing an aqueous fluid in a first microchannel toward a junction, the aqueous fluid comprising a biological reagent;
flowing a hydrophobic fluid that is immiscible with the aqueous fluid in a second microchannel toward the junction, the hydrophobic fluid comprising a fluorosurfactant is linked to an oligoethylene glycol (OEG); and
contacting the aqueous fluid with the hydrophobic fluid at the junction and forming a plurality of plugs at the junction, wherein each plug comprises a subvolume of the aqueous fluid surrounded by the hydrophobic fluid and the subvolume of the aqueous fluid of at least one plug comprises a biological reagent.

### Claim 12 (depends on 11)

The method of claim 11, wherein pressure is applied in the first and second microchannels to induce fluid flow.

### Claim 13 (depends on 12)

The method of claim 12, wherein continuous flow of the hydrophobic fluid is provided in the microchannel after the contacting.

### Claim 14 (depends on 11)

The method of claim 11, wherein the biological reagent is at least one of a cell, a virion, an enzyme, a protein, and a nucleic acid.

### Claim 15 (depends on 11)

The method of claim 11, wherein the fluorosurfactant comprises a hydrophilic head.

### Claim 16 (depends on 11)

The method of claim 11, wherein the fluorosurfactant comprises an oligoethylene glycol terminal.

### Claim 17 (depends on 16)

The method of claim 16, wherein the fluorosurfactant is of the formula:
wherein m is 0, 3, 4, 5, or 6; and n is an integer of no less than 16.

### Claim 18 (depends on 16)

The method of claim 16, where the oligoethylene glycol is linked to a terminal end of the fluorosurfactant.

### Claim 19 (depends on 11)

The method of claim 11, wherein the fluorosurfactant is linked to the oligoethylene glycol (OEG).

### Claim 20

A method of conducting an enzymatic reaction comprising forming a plurality of plugs and conducting the enzymatic reaction in at least one plug, wherein:
each plug comprises an aqueous fluid surrounded by a hydrophobic fluid;
the aqueous fluid of at least one plug comprises an enzyme;
the hydrophobic fluid of each plug comprises a fluorosurfactant; and
the fluorosurfactant is linked to an oligoethylene glycol (OEG).