# US 12,005,454 B2 · Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,005,454 B2 - Document ID: US12005454B2 - Status: Granted - Kind: B2 - Application number: 17/145,858 - Filed: 2021-01-11 - Published: 2021-05-06 - Granted: 2024-06-11 - Earliest claimed: 2014-04-10 - Assignees: 10x Genomics, Inc. - Applicants: 10X Genomics, Inc. - Inventors: Rajiv Bharadwaj; Kevin Ness; Debkishore Mitra; Donald A. Masquelier; Anthony Makarewicz; Christopher Hindson; Benjamin Hindson; Serge Saxonov - Web page: https://patentgrep.com/patent/US12005454B2 ## CPC - 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 3/0241: {Drop counters; Drop formers} - B01L 3/502784: {specially adapted for droplet or plug flow, e.g. digital microfluidics} - B01L 2200/0673: Handling of plugs of fluid surrounded by immiscible fluid - B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions - B01L 2400/049: vacuum - B01L 2400/086: using baffles or other fixed flow obstructions - G01N 35/085: {Flow Injection Analysis} - G01N 2035/00237: {Handling microquantities of analyte, e.g. microvalves, capillary networks} - G01N 2035/1034: {Transferring microquantities of liquid} - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - 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) ## 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. ## Claims ### Claim 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. ### Claim 2 (depends on 1) 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. ### Claim 3 (depends on 2) 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. ### Claim 4 (depends on 1) The microfluidic system of claim 1, wherein the plurality of microcapsules comprises reagents. ### Claim 5 (depends on 4) The microfluidic system of claim 4, wherein the reagents are encapsulated in the plurality of microcapsules. ### Claim 6 (depends on 4) The microfluidic system of claim 4, wherein the reagents are barcode sequences. ### Claim 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. ### Claim 8 (depends on 7) 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. ### Claim 9 (depends on 8) 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. ### Claim 10 (depends on 7) The microfluidic system of claim 7, wherein the agitating component is configured to prevent settling of the plurality of microcapsules. ### Claim 11 (depends on 7) The microfluidic system of claim 7, wherein the plurality of microcapsules comprises reagents. ### Claim 12 (depends on 11) The microfluidic system of claim 11, wherein the reagents are encapsulated in the plurality of microcapsules. ### Claim 13 (depends on 11) The microfluidic system of claim 11, wherein the reagents are barcode sequences. ### Claim 14 (depends on 7) The microfluidic system of claim 7, wherein the agitating component is piezoelectric, acoustic, or mechanical.