# US 11,779,923 B2 · Microfluidic device and a method for provision of double emulsion droplets

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- Number: US 11,779,923 B2
- Document ID: US11779923B2
- Status: Granted
- Kind: B2
- Application number: 16/770,135
- Filed: 2018-12-04
- Published: 2021-08-19
- Granted: 2023-10-10
- Earliest claimed: 2017-12-06
- Assignees: Samplix Aps
- Applicants: Samplix Aps
- Inventors: Thomas Kvist; Esben Bjørn Madsen; Solène Cherre; Marie Just Mikkelsen
- Web page: https://patentgrep.com/patent/US11779923B2

## CPC

- B01L 3/502784: {specially adapted for droplet or plug flow, e.g. digital microfluidics}
- B01F 23/41: Emulsifying
- B01F 23/4105: {Methods of emulsifying}
- B01F 33/3011: using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
- B01F 33/3017: {Mixing chamber}
- B01F 33/813: {mixing simultaneously in two or more mixing receptacles}
- B01L 3/502707: {characterised by the manufacture of the container or its components}
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01F 23/4143: {Microemulsions}
- B01F 23/4144: {Multiple emulsions, in particular double emulsions, e.g. water in oil in water; Three-phase emulsions}
- B01F 2101/23: Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
- B01L 2200/028: Modular arrangements
- B01L 2200/06: Fluid handling related problems
- B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid
- B01L 2200/12: Specific details about manufacturing devices
- B01L 2200/141: Preventing contamination, tampering
- B01L 2300/0681: Filter
- B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01F: MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING (mixing paints B44D3/06)

## Abstract

The present invention relates to a microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, the present invention relates to an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, the present invention relates to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets. The microfluidic device comprises a transfer conduit comprising a first transfer conduit part having a first affinity for water; and a collection conduit comprising a first collection conduit part having a second affinity for water being different from the first affinity for water. A well section and a microfluidic section of the microfluidic device are fixedly connected to each other.

## Claims

### Claim 1

A microfluidic device for production of double emulsion droplets comprising:
a microfluidic section comprising a plurality of microfluidic units; and
a well section comprising a plurality of groups of wells comprising one group of wells for each microfluidic unit;
each microfluidic unit comprising a fluid conduit network comprising:
a plurality of supply conduits comprising a primary supply conduit, a secondary supply conduit, and a tertiary supply conduit;
a transfer conduit comprising a first transfer conduit part having a first affinity for water, wherein the first affinity for water is hydrophobic;
a collection conduit comprising a first collection conduit part having a second affinity for water being different from the first affinity for water, wherein the second affinity for water is hydrophilic;
a first fluid junction providing fluid communication between the primary supply conduit, the secondary supply conduit, and the transfer conduit, the first transfer conduit part extending from the first fluid junction; and
a second fluid junction providing fluid communication between the tertiary supply conduit, the transfer conduit, and the collection conduit, the first collection conduit part extending from the second fluid junction;
each group of wells comprising a plurality of wells comprising a collection well and a plurality of supply wells comprising a primary supply well and a tertiary supply well,
the well section and the microfluidic section being fixedly connected to each other such that each group of wells being fixedly connected to a respective corresponding microfluidic unit, wherein for each group of wells:
the collection well is in fluid communication with the collection conduit of the corresponding microfluidic unit;
the primary supply well is in fluid communication with the primary supply conduit of the corresponding microfluidic unit;
the tertiary supply well is in fluid communication with the tertiary supply conduit of the corresponding microfluidic unit; and
one supply well of the plurality of supply wells is in fluid communication with the secondary supply conduit of the corresponding microfluidic unit,
wherein the microfluidic device is configured for containing liquids in an amount required for the provision of droplets as well as the resulting droplets, and further wherein the microfluidic device is a single use device and produces double emulsion droplets.

### Claim 2 (depends on 1)

The microfluidic device according to claim 1, wherein the first transfer conduit part and the first collection conduit part of each microfluidic unit are configured to retain their respective affinity for water for at least one month of storage from time of provision of the respective conduit parts.

### Claim 3 (depends on 1)

The microfluidic device according to claim 1, wherein the first transfer conduit part or the first collection conduit part of each microfluidic unit is provided by a coating.

### Claim 4 (depends on 3)

The microfluidic device according to claim 3, wherein the microfluidic section comprises a base microfluidic piece providing at least a part of each of: the primary supply conduit of each microfluidic unit; the secondary supply conduit of each microfluidic unit; the tertiary supply conduit of each microfluidic unit; the transfer conduit of each microfluidic unit; the collection conduit of each microfluidic unit; the first fluid junction of each microfluidic unit; and the second fluid junction of each microfluidic unit; and wherein:
the base microfluidic piece is provided in a base material having surface properties corresponding to the first affinity for water, wherein at least a part of the coating providing the first collection conduit part is provided on top of the base material of the base microfluidic piece; or
the base microfluidic piece is provided in a base material having surface properties corresponding to the second affinity for water, wherein at least a part of the coating providing the first transfer conduit part is provided on top of the base material of the base microfluidic piece.

### Claim 5 (depends on 1)

The microfluidic device according to claim 1, wherein, for each microfluidic unit, the cross-sectional area of any opening between any supply conduit and the first fluid junction is smaller than 2500 μm2.

### Claim 6 (depends on 1)

The microfluidic device according to claim 1, wherein, for each microfluidic unit, the cross-sectional area of an opening between the first fluid junction and the transfer conduit is smaller than 2500 μm2.

### Claim 7 (depends on 1)

The microfluidic device according to claim 1, wherein, for each microfluidic unit, the cross-sectional area of an opening between the first fluid junction and the transfer conduit is between 50% and 100% of the cross-sectional area of an opening between the second fluid junction and the collection conduit.

### Claim 8 (depends on 1)

The microfluidic device according to claim 1, wherein the microfluidic section comprises a first planar surface and a capping piece comprising a second planar surface, the first planar surface having a plurality of ramified recesses providing a base part of each fluid conduit network of the microfluidic device, the second planar surface facing the first planar surface and providing a capping part of each fluid conduit network of the microfluidic device.

### Claim 9 (depends on 8)

The microfluidic device according to claim 8, wherein one, more, or all parts of each fluid conduit network form an acute trapezoidal cross section, wherein the longer base edge is provided by the second planar surface of the capping piece.

### Claim 10 (depends on 9)

The microfluidic device according to claim 9, wherein each acute trapezoidal cross section forms an isosceles trapezoidal cross section, wherein the side walls of equal length have a tapering of at least 5 degrees and at most 20 degrees with respect to a normal of either of the parallel base edges.

### Claim 11 (depends on 1)

The microfluidic device according to claim 1, wherein each microfluidic unit comprises:
a primary filter at or within the primary supply conduit;
a secondary filter at or within the secondary supply conduit; and
a tertiary filter at or within the tertiary supply conduit.

### Claim 12 (depends on 1)

The microfluidic device according to claim 1, wherein the first transfer conduit part extends at least 500 μm.

### Claim 13 (depends on 1)

An assembly comprising a receptor and a pressure distribution structure, the receptor being configured to receive and hold the microfluidic device according to claim 1, the pressure distribution structure being configured to supply pressure to the microfluidic device when held by the receptor, the pressure distribution structure comprising:
a plurality of well manifolds comprising a primary well manifold and a tertiary well manifold; a plurality of line pressure regulators comprising a primary line pressure regulator and a tertiary line pressure regulator; and a main manifold;
the primary well manifold being configured to be coupled to each primary supply well of the microfluidic device,
the tertiary well manifold being configured to be coupled to each tertiary supply well of the microfluidic device,
the primary line pressure regulator being coupled to the primary well manifold,
the tertiary line pressure regulator being coupled to the tertiary well manifold,
the main manifold being coupled to each well manifold via the respective line pressure regulators.

### Claim 14 (depends on 1)

A kit comprising:
one or more of the microfluidic device according to claim 1; and
a plurality of fluids configured for use with the microfluidic device according to claim 1, the plurality of fluids comprising: a sample buffer; an oil; and a continuous phase buffer;
the kit comprising an enzyme and nucleotides.

### Claim 15 (depends on 14)

The kit according to claim 14, wherein the density of the oil is higher than the density of the sample buffer.

### Claim 16 (depends on 1)

A method for providing double emulsion droplets, the method comprising use of any of:
the microfluidic device according to claim 1; or for the provision of double emulsion droplets, and comprising:
providing a first fluid to the primary supply well of a first group of wells;
providing a second fluid to the one supply well of the first group of wells, which supply well is in fluid communication with the secondary supply conduit of the corresponding microfluidic unit;
providing a third fluid to the tertiary supply well of the first group of wells; and providing individual pressure differences between each of the respective supply wells of the first group of wells and the collection well of the first group of wells, such that the pressure within each of the individual supply wells of the first group of wells is higher than within the collection well of the first group of wells.

### Claim 17 (depends on 1)

A method for manufacturing a microfluidic device according to claim 1, the method comprising fixing the well section and the microfluidic section to each other, such that fluid communication is provided between the individual wells of each group of wells via the corresponding respective microfluidic units.

### Claim 18 (depends on 1)

The microfluidic device according to claim 1, wherein the first affinity for water of said transfer conduit part is hydrophobic and wherein the second affinity for water of said first collection conduit part is hydrophilic and wherein said droplets are double-emulsion droplets.

### Claim 19 (depends on 16)

The method according to claim 16, wherein the one supply well is the primary supply well, the secondary supply well or the tertiary supply well in fluid communication with the secondary supply conduit.