System for generating droplets—instruments and cassette

• US 9,636,682 B2
• Filed: 11/14/2016
• Issued: 05/02/2017
• Est. Priority Date: 09/23/2008
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Hiddessen, Amy L., Ness, Kevin D., Hindson, Benjamin J., Masquelier, Donald A.
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US15/351,335
• Publication Number:
  US 20170056886A1
• Time in Patent Office: 169 Days
• CPC Class Codes: B01F 23/41 Emulsifying
• CPC Class Codes: B01F 33/3011 using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/088 Channel loops
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 3/0241 Drop counters; Drop formers making arrays for combinatorial libraries B01J19/0046; automation of dispensing for analysis G01N35/10
• CPC Class Codes: B01L 3/502 : with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• 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 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 3/52 : Containers specially adapted for storing or dispensing a reagent B01L3/02 takes precedence; containers for medical or pharmaceutical purposes A61J1/00; containers in general B65D; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: G01N 15/1404 : Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1433 : using image recognition
• CPC Class Codes: G01N 15/1434 : Optical arrangements
• CPC Class Codes: G01N 15/1436 : the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
• CPC Class Codes: G01N 15/1459 : the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1415 : Control of particle position

Abstract

System, including methods, apparatus, and kits, for forming emulsions. An exemplary system may comprise a device including a sample well configured to receive sample-containing fluid, a continuous-phase well configured to receive continuous-phase fluid, and a droplet well. The device also may include a channel network having a first channel, a second channel, and a third channel that meet one another in a droplet-generation region. The system also may comprise a holder for the device. The system further may comprise an instrument configured to operatively receive an assembly including the device and the holder and to drive sample-containing fluid from the sample well to the droplet-generation region via the first channel, continuous-phase fluid from the continuous-phase well to the droplet-generation region via the second channel, and sample-containing droplets from the droplet-generation region to the droplet well via the third channel.

Claims

1. A system for generating droplets, comprising:

a device including a sample well configured to receive sample-containing fluid, a continuous-phase well configured to receive continuous-phase fluid, and a droplet well, the device also including a channel network having a first channel, a second channel, and a third channel that meet one another in a droplet-generation region;

a holder for the device; and

an instrument configured to operatively receive an assembly including the device and the holder and to drive sample-containing fluid from the sample well to the droplet-generation region via the first channel, continuous-phase fluid from the continuous-phase well to the droplet-generation region via the second channel, and sample-containing droplets from the droplet-generation region to the droplet well via the third channel.

2. The system of claim 1, further comprising a gasket configured to engage a rim of at least one of the wells.

3. The system of claim 2, wherein the gasket also is configured to engage the holder.

4. The system of claim 2, wherein the gasket is configured to be attached directly to the holder.

5. The system of claim 3, wherein the gasket defines an aperture, and wherein the holder has a projection configured to be received in the aperture.

6. The system of claim 1, wherein the holder has a larger footprint than the device.

7. The system of claim 1, wherein the device has a planar base portion and a plurality of tubular protrusions that project upwardly from the base portion, and wherein the device is configured to be captured by the holder via the base portion.

8. The system of claim 1, wherein the instrument forms a depression in which the holder is configured to be operatively received.

9. The system of claim 1, wherein holder is adjustable between (a) an open configuration that allows the device to be placed into and removed from the holder and (b) a closed configuration that prevents removal of the device from the holder.

10. The system of claim 1, wherein the holder defines at least one opening, and wherein the instrument includes at least one pin configured to enter the at least one opening when the assembly is operatively received by the instrument.

11. The system of claim 1, wherein instrument is configured to detect that the holder is properly positioned in the instrument.

12. The system of claim 11, wherein the instrument is configured to detect that the holder is properly positioned in the instrument using an electrode.

13. The system of claim 1, further comprising a gasket configured to be placed onto the device, wherein the instrument is configured to optically detect whether the gasket is present on the device when the assembly is operatively received by the instrument.

14. A system for generating droplets, comprising:

a device including a row of sample wells each configured to receive sample-containing fluid, a row of continuous-phase wells each configured to receive continuous-phase fluid, and a row of droplet wells, the device also including a corresponding channel network for each sample well, the channel network including a droplet-generation region and fluidically connecting the sample well to one of the continuous-phase wells and one of the droplet wells;

a holder for the device;

a gasket configured to be attached directly to the holder, such that the gasket extends over each sample well, each continuous-phase well, and each droplet well; and

an instrument configured to

(a) receive an assembly including the device, the holder, and the gasket,

(b) engage the gasket with a manifold, and

(c) apply positive pressure and/or negative pressure to the device via the manifold, such that sample-containing fluid flows from each sample well to the corresponding droplet-generation region, continuous-phase fluid flows from each continuous-phase well to the corresponding droplet-generation region, and sample-containing droplets flow from each droplet-generation region to the corresponding droplet well.

15. The system of claim 14, wherein the instrument is configured to apply negative pressure to each droplet well of the row of droplet wells to drive formation of sample-containing droplets at each droplet-generation region.

16. The system of claim 14, wherein the gasket defines a plurality of apertures, and wherein the holder has a plurality of projections configured to be received in the plurality of apertures to directly attach the gasket to the holder.

17. The system of claim 14, wherein the device has a planar base portion and a plurality of tubular protrusions that project upwardly from the planar base portion, and wherein the device is configured to be captured in the holder via the planar base portion.

18. The system of claim 14, wherein the instrument is configured to detect that the holder has been received in the instrument.

19. The system of claim 14, wherein the instrument is configured to detect whether the gasket is present after the assembly has been received by the instrument.

20. A system to form an emulsion, comprising:

a device including a sample well configured to receive sample-containing fluid, a continuous-phase well configured to receive continuous-phase fluid, and a droplet well, the device also including a channel network including a droplet-generation region, the channel network fluidically connecting the wells to one another;

a holder configured to receive the device; and

an instrument configured to

(a) receive an assembly including the device and the holder,

(b) detect that the holder has been received, and

(c) create a pressure differential, such that sample-containing fluid flows from the sample well to the droplet-generation region, continuous-phase fluid flows from the continuous-phase well to the droplet-generation region, and sample-containing droplets flow from the droplet-generation region to the droplet well.

21. The system of claim 20, further comprising a gasket configured to be placed on the device, wherein the instrument is configured to detect whether the gasket is present on the device after the assembly has been received.