Instrument systems for integrated sample processing

• US 9,975,122 B2
• Filed: 11/05/2015
• Issued: 05/22/2018
• 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:
  US14/934,044
• Publication Number:
  US 20160121325A1
• Time in Patent Office: 929 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. An integrated system for processing samples comprised of various fluids for analysis, the system comprising:

(a) a microfluidic device including a plurality of channel networks for partitioning the samples into partitioned samples, the plurality of channel networks being connected to a plurality of inlet and outlet reservoirs;

(b) a holder including a closeable lid hingedly coupled thereto,

wherein in a closed configuration, the closeable lid secures the microfluidic device in the holder, and

wherein in an open configuration, the closeable lid comprises a stand orienting the microfluidic device at an angle to facilitate recovery of the partitioned samples; and

(c) 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 plurality of channel networks.

2. The integrated system of claim 1, wherein the angle at which the microfluidic device is oriented by the closeable lid ranges from about 20 degrees to about 70 degrees.

3. The integrated system of claim 1, wherein the angle at which the microfluidic device is oriented by the closeable lid is approximately 45 degrees.

4. The integrated system of claim 1, wherein the instrument comprises:

(a) a retractable tray supporting and seating the holder, and slidable into and out of the instrument;

(b) a depressible manifold assembly configured to be actuated and lowered to the microfluidic device and to sealaby interface with the plurality of inlet and outlet reservoirs;

(c) at least one fluid drive component configured to apply the pressure differential between the plurality of inlet and outlet reservoirs; and

(d) a controller configured to operate the at least one fluid drive component to apply the pressure differential depending on a mode of operation or according to preprogrammed instructions.

5. The integrated system of claim 1, wherein at least a portion of one or more of the various fluids comprises reagents,

wherein at least one of the plurality of channel networks comprises a plurality of interconnected fluid channels fluidly communicated at a first channel junction, at which an aqueous phase containing at least one of the reagents is combined with a stream of a non-aqueous fluid to partition the aqueous phase into discrete droplets within the non-aqueous fluid, and

wherein the discrete droplets are stored in at least one outlet reservoir of the plurality of inlet and outlet reservoirs, or stored in at least one product storage vessel.

6. The integrated system of claim 5, wherein the plurality of interconnected fluid channels comprises a microfluidic structure having intersecting fluid channels fabricated into a monolithic component part.

7. The integrated system of claim 4, further comprising a gasket coupled to the holder and including a plurality of apertures,

wherein, when the closeable lid is in the closed configuration, the gasket is positioned between the plurality of inlet and outlet reservoirs and the depressible manifold assembly to provide a sealable interface, and

wherein the plurality of apertures allows pressure communication between at least one of the outlet and the inlet reservoirs and the at least one fluid drive component.

8. The integrated system of claim 4, further comprising springs to bias the depressible manifold assembly in a raised position, and a servo motor to actuate and lower the depressible manifold assembly.

9. The integrated system of claim 1, further comprising at least one monitoring component interfaced with at least one of the plurality of channel networks and configured to observe and monitor characteristics and properties of the at least one of the plurality of channel networks and fluids flowing therein, wherein the at least one monitoring component is selected from the group consisting of:

a temperature sensor, a pressure sensor, and a humidity sensor.

10. The integrated system of claim 1, wherein at least one channel of the plurality of channel networks comprises a channel segment that widens and controls flow by breaking capillary forces acting to draw aqueous fluids into the at least one channel.

11. The integrated system of claim 10, further comprising a passive check valve comprising the channel segment.

12. The integrated system of claim 1, wherein at least one of the plurality of channel networks comprises:

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

a second channel segment fluidly coupled to a source of a sample of the samples, the first and second channel segments fluidly connected at a first channel junction;

a third channel segment connected to the first and second channel segments at the first channel junction;

a fourth channel segment connected to the third channel segment at a second channel junction and connected to a source of partitioning fluid; and

a fifth channel segment fluidly coupled to the second channel junction and connected to a channel outlet,

wherein the instrument is coupled to at least one of the first, second, third, fourth, and fifth channel segments, and is configured to drive flow of the barcode reagents and the sample into the first channel junction to form a reagent mixture in the third channel segment and to drive flow of the reagent mixture and the partitioning fluid into the second channel junction to form droplets of the reagent mixture in a stream of partitioning fluid within the fifth channel segment.

16. The integrated system of claim 1, wherein each of the plurality of channel networks are parallel with one another.

17. The integrated system of claim 1, wherein the angle at which the microfluidic device is oriented by the closeable lid ranges from about 30 degrees to about 60 degrees.

18. The integrated system of claim 1, wherein the angle at which the microfluidic device is oriented by the closeable lid ranges from about 40 degrees to about 50 degrees.

13. A holder assembly, comprising:

(a) a holder body configured to receive a microfluidic device, the microfluidic device including a plurality of parallel channel networks for partitioning various fluids; and

(b) a closeable lid hingedly coupled to the holder body,

wherein in a closed configuration, the closeable lid secures the microfluidic device in the holder body, and in an open configuration, the closeable lid comprises a stand to orient the microfluidic device at an angle to facilitate recovery of partitioned fluids without spilling the partitioned fluids.

14. The holder assembly of claim 13, wherein the angle at which the microfluidic device is oriented by the closeable lid ranges from about 20 degrees to about 70 degrees.

15. The holder assembly of claim 13, wherein the angle at which the microfluidic device is oriented by the closeable lid is approximately 45 degrees.