# US 11,999,932 B2 · Microfluidic control

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- Number: US 11,999,932 B2
- Document ID: US11999932B2
- Status: Granted
- Kind: B2
- Application number: 17/036,652
- Filed: 2020-09-29
- Published: 2021-01-28
- Granted: 2024-06-04
- Earliest claimed: 2018-04-03
- Assignees: Emulate, Inc.
- Applicants: EMULATE, Inc.
- Inventors: Christopher David Hinojosa; Grace Ahn
- Web page: https://patentgrep.com/patent/US11999932B2

## CPC

- C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)}
- F04B 19/006: {Micropumps (F04B43/043 and F04B43/095 take precedence)}
- B01L 3/505: {Flexible containers without fluid transport within}
- C12M 23/26: {flexible (flexible containers for laboratory use B01L3/505)}
- C12M 25/02: {Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00)}
- C12M 29/00: {Means for introduction, extraction or recirculation of materials, e.g. pumps (pumps per se F04B)}
- C12M 41/18: {Heat exchange systems, e.g. heat jackets or outer envelopes}
- F04B 23/02: having reservoirs
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/50273: {characterised by the means or forces applied to move the fluids}
- B01L 2400/0481: squeezing of channels or chambers
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- F04B: POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS (machines for liquids, or pumps, of rotary-piston or oscillating-piston type F04C; non-positive-displacement pumps F04D; pumping of fluid by direct contact of another fluid or by using inertia of fluid to be pumped F04F)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

The present invention is related to the field of fluidic devices, and in particular, microfluidic cell culture systems. The present invention provides pumping, recirculation and sampling for a microfluidic device or devices. The present invention provides solutions to the control of microfluidics for both terrestrial and space applications, including the control over the movement of fluids in zero gravity or microgravity.

## Claims

### Claim 1

A method of moving liquid, comprising:
a) providing an assembly, comprising one or more liquid reservoirs contained within a container, said container in fluidic communication with e a first working fluid reservoir and a second working fluid reservoir, said first working fluid reservoir comprising a first working fluid, said second working fluid reservoir comprising a second working fluid, said container further comprising a portion of said second working fluid, said one or more liquid reservoirs in fluidic communication with one or more fluidic devices, and comprising liquid, wherein there is no mixing of said working fluid with said liquid; and
b) introducing said first working fluid from said first working fluid reservoir into said container under conditions that said pressure is imparted on said second working fluid such that liquid in said one or more reservoirs is displaced and flows into said one or more fluidic devices, thereby moving said liquid.

### Claim 2 (depends on 1)

The method of claim 1, wherein said container is rigid or flexible.

### Claim 3 (depends on 1)

The method of claim 1, wherein said container is collapsible.

### Claim 4 (depends on 1)

The assembly of claim 1, wherein said container is insulated.

### Claim 5 (depends on 1)

The method of claim 1, wherein said container is sealed such that said working fluid can only enter or exit the container through conduits.

### Claim 6 (depends on 1)

The method of claim 1, wherein said one or more reservoirs are flexible.

### Claim 7 (depends on 1)

The method of claim 1, wherein said one or more reservoirs comprise at least one deformable portion.

### Claim 8 (depends on 1)

The method of claim 1, wherein said first working fluid comprises gas.

### Claim 9 (depends on 1)

The method of claim 1, further comprising cooling said first or second working fluid, or both.

### Claim 10 (depends on 9)

The method of claim 9, wherein said working fluid is cooled with a cooling conduit comprising a cooling fluid, wherein said cooling fluid does not mix with said working fluid or with said liquid.

### Claim 11 (depends on 10)

The method of claim 10, wherein said cooling conduit comprises stainless steel tubing.

### Claim 12 (depends on 1)

The method of claim 1, wherein said liquid is heated after it is displaced but prior to it flowing into said one or more fluidic devices.

### Claim 13

A method of introducing reagent into microfluidic devices comprising:
a) providing an assembly, comprising one or more flexible cell culture media reservoirs and one or more flexible reagent reservoirs contained within a container, said container in fluidic communication with a pump and a working fluid reservoir and comprising a working fluid, said flexible cell culture reservoirs in fluidic communication with one or more microfluidic devices comprising cells on a surface and comprising cell culture media, said one or more flexible reagent reservoirs in fluidic communication with one or more microfluidic devices and comprising reagent, wherein there is no contact or mixing of said working fluid with said cell culture media;
b) pumping said working fluid into said container with said pump under conditions that the culture media in the flexible reservoirs is displaced and flows into said one or more microfluidic devices, thereby perfusing said cells, wherein reagent in said flexible reagent reservoirs is not displaced; and
c) pumping said working fluid into said container with said pump under conditions that the reagent in at least one flexible reagent reservoir is displaced and flows into said one or more microfluidic devices, thereby introducing reagent into said one or more microfluidic devices.

### Claim 14 (depends on 13)

The method of claim 13, wherein said assembly further comprises valves to control the flow of said culture media out of said flexible cell culture media reservoirs and valves to control the flow of reagent out of said flexible reagent reservoirs.

### Claim 15 (depends on 14)

The method of claim 14, wherein a valve prevents reagent in said flexible reagent reservoir from being displaced in step 2).

### Claim 16 (depends on 14)

The method of claim 14, wherein a valve permits reagent in at least one of said flexible reagent reservoir to be displaced in step 3).

### Claim 17 (depends on 13)

The method of claim 13, wherein said one or more microfluidic devices are outside said container.

### Claim 18 (depends on 13)

The method of claim 13, wherein said one or more microfluidic devices are housed in a manifold.

### Claim 19 (depends on 13)

The method of claim 13, wherein said surface is a microfluidic channel or portion thereof.

### Claim 20 (depends on 13)

The method of claim 13, wherein said surface is a membrane.

### Claim 21 (depends on 13)

The method of claim 13, wherein said container is rigid or flexible.

### Claim 22 (depends on 13)

The method of claim 13, wherein said container is collapsible.

### Claim 23 (depends on 13)

The method of claim 13, wherein said container is insulated.

### Claim 24 (depends on 13)

The method of claim 13, wherein said container is sealed such that said working fluid can only enter or exit the container through conduits.

### Claim 25 (depends on 13)

The method of claim 13, further comprising cooling said working fluid.

### Claim 26 (depends on 25)

The method of claim 25, wherein said working fluid is cooled with a cooling conduit comprising a cooling fluid, wherein said cooling fluid does not mix with said working fluid or with said liquid.

### Claim 27 (depends on 26)

The method of claim 26, wherein said cooling conduit comprises stainless steel tubing.

### Claim 28 (depends on 13)

The method of claim 13, wherein said liquid is heated after it is displaced but prior to it flowing into said one or more fluidic devices.

### Claim 29

A method of collecting samples from microfluidic devices, comprising:
a) providing an assembly, comprising one or more reservoirs contained within a container, said container in fluidic communication with a pump and a working fluid reservoir and comprising a working fluid, said reservoirs in fluidic communication with one or more microfluidic devices and comprising cell culture media, said one or more microfluidic devices in fluidic communication with a sampling conduit and comprising cells on a surface, wherein there is no mixing of said working fluid with said cell culture media;
b) pumping said working fluid into said container with said pump under conditions that the culture media in the reservoirs is displaced and flows into said one or more microfluidic devices, thereby causing fluid to exit said one or more microfluidic devices and enter said sampling conduit; and
c) collecting samples from said sampling conduit.

### Claim 30 (depends on 29)

The method of claim 29, further comprising introducing gas into said sampling conduit to separate said fluid into discrete volumes.

### Claim 31 (depends on 29)

The method of claim 29, wherein said container is rigid or flexible.

### Claim 32 (depends on 29)

The method of claim 29, wherein said container is collapsible.

### Claim 33 (depends on 29)

The method of claim 29, wherein said container is insulated.

### Claim 34 (depends on 29)

The method of claim 29, wherein said container is sealed such that said working fluid can only enter or exit the container through conduits.

### Claim 35 (depends on 29)

The method of claim 29, further comprising cooling said working fluid.

### Claim 36 (depends on 35)

The method of claim 35, wherein said working fluid is cooled with a cooling conduit comprising a cooling fluid, wherein said cooling fluid does not mix with said working fluid or with said liquid.

### Claim 37 (depends on 36)

The method of claim 36, wherein said cooling conduit comprises stainless steel tubing.

### Claim 38 (depends on 29)

The method of claim 29, wherein said liquid is heated after it is displaced but prior to it flowing into said one or more fluidic devices.

### Claim 39

A method of recirculating culture media through microfluidic devices without reversing the direction of fluid flow, comprising:
a) providing an assembly, comprising one or more reservoirs contained within a container, said container in fluidic communication with a pump and a working fluid reservoir and comprising a working fluid, said reservoirs in fluidic communication with one or more microfluidic devices and comprising cell culture media, said one or more microfluidic devices comprising cells on a surface and inlet and outlet ports, said inlet and outlet ports in fluidic communication with a recirculation pathway, wherein there is no mixing of said working fluid with said cell culture media; and
b) pumping said working fluid into said container with said pump under conditions that the culture media in the reservoirs is displaced and flows into said one or more microfluidic devices in a direction, thereby causing fluid to exit said outlet port of said one or more microfluidic devices and enter said recirculation pathway, moving in the direction of said inlet port of said one or more microfluidic devices.

### Claim 40 (depends on 39)

The method of claim 39, wherein said pumping causes said fluid moving in the direction of said inlet port to reach said inlet port, thereby recirculating said culture media without reversing the direction of fluid flow.

### Claim 41 (depends on 39)

The method of claim 39, wherein said one or more microfluidic devices are housed on a manifold.

### Claim 42 (depends on 41)

The method of claim 41, wherein said recirculation pathway is positioned on said manifold.

### Claim 43 (depends on 39)

The method of claim 39, wherein said container is rigid or flexible.

### Claim 44 (depends on 39)

The method of claim 39, wherein said container is collapsible.

### Claim 45 (depends on 39)

The method of claim 39, wherein said container is insulated.

### Claim 46 (depends on 39)

The method of claim 39, wherein said container is sealed such that said working fluid can only enter or exit the container through conduits.

### Claim 47 (depends on 39)

The method of claim 39, further comprising cooling said working fluid.

### Claim 48 (depends on 47)

The method of claim 47, wherein said working fluid is cooled with a cooling conduit comprising a cooling fluid, wherein said cooling fluid does not mix with said working fluid or with said liquid.

### Claim 49 (depends on 48)

The method of claim 48, wherein said cooling conduit comprises stainless steel tubing.

### Claim 50 (depends on 39)

The method of claim 39, wherein said liquid is heated after it is displaced but prior to it flowing into said one or more fluidic devices.