Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same

Patent metadata

Field	Value
Publication number	US-12005454-B2
Application number	US-202117145858-A
Country	US
Kind code	B2
Filing date	Jan 11, 2021
Priority date	Apr 10, 2014
Publication date	Jun 11, 2024
Grant date	Jun 11, 2024

Abstract

The disclosure provides devices, systems and methods for the generation of encapsulated reagents and the partitioning of encapsulated reagents for use in subsequent analyses and/or processing, such as in the field of biological analyses and characterization.

First claim

The invention claimed is:

1. A microfluidic system, comprising:
a first fluid source fluidly connected to a first channel and comprising a first fluid that comprises an aqueous fluid and a plurality of microcapsules;
an agitating component disposed to agitate the first fluid source;
a second fluid source fluidly connected to a second channel and comprising a second fluid that is immiscible with the aqueous fluid;
a droplet generation junction fluidly connected to a third channel; and
a flow control system that comprises a first independently controlled pressure source that causes the first fluid from the first fluid source to flow along the first channel and a second independently controlled pressure source that causes the second fluid from the second fluid source to flow along the second channel, such that the microcapsules and the second fluid meet at the droplet generation junction to generate a plurality of droplets comprising the microcapsules from the plurality of microcapsules, which plurality of droplets flow along the third channel.

2. The microfluidic system of claim 1 , further comprising a third fluid source fluidly connected to the droplet generation junction through a fourth channel, wherein the third fluid source comprises a third fluid comprising an additional aqueous fluid.

3. The microfluidic system of claim 2 , further comprising a third independently controlled pressure source, wherein the first channel and the fourth channel meet at an additional junction upstream of the droplet generation junction, wherein the third independently controlled pressure source causes the third fluid from the third fluid source to flow along the fourth channel to the additional junction, thereby bringing the first fluid and the third fluid in contact to form a mixture comprising the aqueous fluid from the first fluid source and the additional aqueous fluid from the third fluid source.

4. The microfluidic system of claim 1 , wherein the plurality of microcapsules comprises reagents.

5. The microfluidic system of claim 4 , wherein the reagents are encapsulated in the plurality of microcapsules.

6. The microfluidic system of claim 4 , wherein the reagents are barcode sequences.

7. A microfluidic system, comprising:
a first fluid source fluidly connected to a first channel and comprising a first fluid that comprises an aqueous fluid and a plurality of microcapsules;
an agitating component disposed to agitate the first fluid source;
a second fluid source fluidly connected to a second channel and comprising a second fluid that is immiscible with the aqueous fluid;
a droplet generation junction fluidly connected to a third channel; and
a flow control system that:
(i) causes the first fluid from the first fluid source to flow along the first channel, and
(ii) causes the second fluid from the second fluid source to flow along the second channel, such that the microcapsules and the second fluid meet at the droplet generation junction to generate a plurality of droplets comprising the microcapsules from the plurality of microcapsules, which plurality of droplets flow along the third channel.

8. The microfluidic system of claim 7 , further comprising a third fluid source fluidly connected to the droplet generation junction through a fourth channel, wherein the third fluid source comprises a third fluid comprising an additional aqueous fluid.

9. The microfluidic system of claim 8 , wherein the first channel and the fourth channel meet at an additional junction upstream of the droplet generation junction, wherein the flow control system causes the third fluid from the third fluid source to flow along the fourth channel to the additional junction, thereby bringing the first fluid and the third fluid in contact to form a mixture comprising the aqueous fluid from the first fluid source and the additional aqueous fluid from the third fluid source.

10. The microfluidic system of claim 7 , wherein the agitating component is configured to prevent settling of the plurality of microcapsules.

11. The microfluidic system of claim 7 , wherein the plurality of microcapsules comprises reagents.

12. The microfluidic system of claim 11 , wherein the reagents are encapsulated in the plurality of microcapsules.

13. The microfluidic system of claim 11 , wherein the reagents are barcode sequences.

14. The microfluidic system of claim 7 , wherein the agitating component is piezoelectric, acoustic, or mechanical.

Assignees

• 10X Genomics Inc

Inventors

• Bharadwaj Rajiv
• Ness Kevin
• Mitra Debkishore
• Masquelier Donald A
• Makarewicz Anthony
• Hindson Christopher
• Hindson Benjamin
• Saxonov Serge

Classifications

• B01L2400/086

  using baffles or other fixed flow obstructions · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• B01L2200/0673

  Handling of plugs of fluid surrounded by immiscible fluid · CPC title

• B01L3/502784

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title

• B01L3/0241 Primary

  Drop counters; Drop formers · CPC title