Instrument systems for integrated sample processing

• US 10,245,587 B2
• Filed: 04/20/2018
• Issued: 04/02/2019
• Est. Priority Date: 11/05/2014
• Status: Active Grant

• Current Assignee: 10X Genomics Incorporated
• Original Assignee: 10X Genomics Incorporated
• Inventors: Masquelier, Donald A., Hindson, Benjamin, Ness, Kevin, Bharadwaj, Rajiv
• Primary Examiner(s): Hobbs, Michael L
• Application Number:
  US15/958,391
• Publication Number:
  US 20180236443A1
• Time in Patent Office: 347 Days
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/043 Hinged closures
• CPC Class Codes: B01L 2300/0609 Holders integrated in container to position an object
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0605 check valves
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: F16K 2099/0084 Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0057 the fluid being the circulating fluid itself, e.g. check valves

Abstract

An integrated system for processing and preparing samples for analysis may include a microfluidic device including a plurality of parallel channel networks for partitioning the samples including various fluids, and connected to a plurality of inlet and outlet reservoirs, at least a portion of the fluids comprising reagents, a holder including a closeable lid hingedly coupled thereto, in which in a closed configuration, the lid secures the microfluidic device in the holder, and in an open configuration, the lid is a stand orienting the microfluidic device at a desired angle to facilitate recovery of partitions or droplets from the partitioned samples generated within the microfluidic device, and an instrument configured to receive the holder and apply a pressure differential between the plurality of inlet and outlet reservoirs to drive fluid movement within the channel networks.

Claims

1. A system for processing samples, comprising:

a microfluidic device including a plurality of channel networks configured to partition said samples into partitioned samples, wherein a channel network of said plurality of channel networks is connected to an inlet reservoir and an outlet reservoir;

an instrument configured to interface with said microfluidic device and apply a pressure differential between said inlet reservoir and said outlet reservoir to subject fluid within said channel network to movement; and

a holder configured to receive said microfluidic device; and

a rotatable body coupled to said holder, wherein said rotatable body is configured to rotate from a closed configuration to an open configuration, or vice versa,

wherein said holder is configured to (i) secure said microfluidic device and permit said instrument to interface with said microfluidic device to apply said pressure differential when said rotatable body is in said closed configuration, and (ii) permit said microfluidic device to be inserted into or removed from said holder when said rotatable body is in said open configuration.

2. The system of claim 1, wherein in said open configuration, said rotatable body is configured to support said holder against a surface.

3. The system of claim 1, wherein said instrument comprises:

(a) a tray configured to support and retain said holder, which tray is disposable inside or outside said instrument;

(b) a manifold assembly configured to be actuated to interface with said microfluidic device;

(c) at least one fluid drive component configured to apply said pressure differential between said inlet reservoir and said outlet reservoir; and

(d) a controller configured to direct application of said pressure differential.

4. The system of claim 3, further comprising a gasket comprising a plurality of apertures, wherein when said rotatable body is in said closed configuration, said gasket is positioned between said microfluidic device and said manifold assembly to provide a sealable interface, and wherein said plurality of apertures is configured to permit fluid communication between said at least one fluid drive component and at least one of said outlet reservoir and said inlet reservoir.

5. The system of claim 3, further comprising a biasing unit that is configured to bias said manifold assembly in a raised position and an actuator that is configured to lower said manifold assembly.

6. The system of claim 1, wherein said channel network comprises a plurality of interconnected fluid channels connected at a channel junction, wherein said channel junction is configured to combine a first fluid containing at least a subset of said samples with a stream of a second fluid immiscible with said first fluid, to partition said at least said subset of said samples into discrete droplets within said second fluid, to thereby provide at least a subset of said partitioned samples within said discrete droplets, which discrete droplets are stored in said outlet reservoir or a storage vessel.

7. The system of claim 6, wherein said plurality of interconnected fluid channels is part of a monolithic microfluidic structure having intersecting fluid channels.

8. The system of claim 1, further comprising at least one monitoring component interfaced with said channel network, wherein said at least one monitoring component is configured to observe or monitor one or more characteristics or properties of said at least one of said plurality of channel networks and fluids flowing therein.

9. The system of claim 1, wherein at least one channel of said channel network comprises a channel segment that widens, wherein said channel segment is configured to control flow by breaking capillary forces acting to draw a fluid into said at least one channel.

10. The system of claim 1, wherein at least one channel of said channel network comprises a passive check valve.

11. The system of claim 1, wherein said channel network comprises:

a first channel segment fluidly connected to a source of barcode reagents;

a second channel segment fluidly connected to a source of a sample of said samples, wherein said first channel segment and said second channel segment are fluidly connected to a first channel junction;

a third channel segment and a fourth channel segment, wherein said third channel segment is fluidly connected to said first channel junction, wherein said fourth channel segment is fluidly connected to a source of partitioning fluid, and wherein said third channel segment and said fourth channel segment are fluidly connected to a second channel junction; and

a fifth channel segment fluidly connected to said second channel junction,

wherein said instrument is configured to interface with said channel network to (i) drive flow of said barcode reagents and said sample into said first channel junction to form a reagent mixture comprising said barcode reagents and said sample in said third channel segment, and (ii) drive flow of said reagent mixture and said partitioning fluid into said second channel junction to form droplets comprising said reagent mixture in a stream of partitioning fluid within said fifth channel segment.

12. The system of claim 1, wherein said plurality of channel networks are substantially parallel to one another.

13. The system of claim 1, wherein an additional channel network of said plurality of channel networks is connected to an additional inlet reservoir and an additional outlet reservoir, which additional inlet reservoir and additional outlet reservoir are separate from said inlet reservoir and outlet reservoir.

14. A holder assembly, comprising:

a holder configured to receive a microfluidic device comprising a plurality of channel networks for partitioning samples; and

a rotatable body coupled to said holder, wherein said rotatable body is configured to rotate from a closed configuration to an open configuration, or vice versa, and wherein said rotatable body is configured to (i) permit said microfluidic device to be secured in said holder when said rotatable body is in said closed configuration, and (ii) permit said microfluidic device to be inserted into or removed from said holder when said rotatable body is in said open configuration.

15. The holder assembly of claim 14, further comprising a gasket coupled to said rotatable body.

16. The holder assembly of claim 15, wherein said gasket is removable from said rotatable body.

17. The holder assembly of claim 15, wherein said gasket comprises securing features configured to mate with complementary features on said rotatable body.

18. The holder assembly of claim 15, wherein said gasket comprises a plurality of apertures configured to be aligned with an inlet reservoir or an outlet reservoir of a channel network of said plurality of channel networks when said microfluidic device is secured in said holder and said rotatable body is in said closed configuration.

19. The holder assembly of claim 14, further comprising said microfluidic device secured in said holder.

20. The holder assembly of claim 19, wherein said plurality of channel networks are substantially parallel to one another.

21. The holder assembly of claim 14, further comprising a gasket coupled to said microfluidic chip.

22. The hold assembly of claim 21, wherein said gasket is removable from said microfluidic chip.