Droplet-based assay system

Patent metadata

Field	Value
Publication number	US-11633739-B2
Application number	US-202017137274-A
Country	US
Kind code	B2
Filing date	Dec 29, 2020
Priority date	Sep 23, 2008
Publication date	Apr 25, 2023
Grant date	Apr 25, 2023

Abstract

System for performing a flow-based assay. The system may comprise a droplet generator to produce an emulsion including droplets in a carrier fluid. The system also may comprise a thermocycler including two or more temperature-controlled zones and also including a channel connected to the droplet generator for receiving the emulsion. The channel may form a single-pass continuous fluid route traversing the temperature-controlled zones multiple times, such that droplets passing through the channel are thermally cycled. The system further may comprise a detection station downstream from the thermocycler and configured to detect a signal from the droplets after such droplets have been thermally cycled by passing through the channel.

First claim

We claim:

1. A system for performing a flow-based assay, the system comprising:
a droplet generator to produce an emulsion including droplets in a carrier fluid;
a thermocycler including two or more temperature-controlled zones and also including a channel connected to the droplet generator for receiving the emulsion, the channel forming a single-pass continuous fluid route traversing the temperature-controlled zones multiple times, such that droplets passing through the channel are thermally cycled; and
a detection station downstream from the thermocycler and configured to detect a signal from the droplets after such droplets have been thermally cycled by passing through the channel.

2. The system of claim 1 , further comprising a disposable cartridge configured to be removably coupled to the thermocycler, the disposable cartridge containing one or more reagents.

3. The system of claim 2 , wherein the disposable cartridge contains one or more samples.

4. The system of claim 3 , wherein the disposable cartridge includes the droplet generator.

5. The system of claim 1 , wherein the droplet generator is configured to generate a plurality of emulsions separated from one another by spacer liquid.

6. The system of claim 5 , wherein the spacer liquid is labeled to be optically distinguishable from the carrier fluid.

7. The system of claim 1 , further comprising at least one pump configured to drive fluid through the droplet generator.

8. The system of claim 1 , further comprising at least one pump configured to drive the droplets through the channel and the detection station.

9. The system of claim 1 , wherein the thermocycler includes at least three temperature-controlled zones having independently controllable temperatures.

10. The system of claim 1 , wherein the fluid route is helical.

11. The system of claim 10 , wherein the fluid route extends about a central axis, and wherein at least one of the temperature-controlled zones varies in size along the central axis.

12. The system of claim 1 , wherein the thermocycler includes a thermoelectric cooler.

13. The system of claim 12 , wherein the thermocycler includes a thermal core, and wherein the thermoelectric cooler is configured to transfer heat between the thermal core and at least one of the temperature-controlled zones.

14. The system of claim 1 , wherein the thermocycler includes a thermal core and a plurality of segments attached to the thermal core and defining the temperature-controlled zones.

15. The system of claim 1 , wherein the channel is provided by tubing wrapped around a core of the thermocycler.

16. The system of claim 1 , wherein the detection station is an optical detection station configured to detect fluorescence of at least one color from the droplets.

17. The system of claim 16 , wherein the optical detection station is configured to perform multi-color fluorescence detection.

18. The system of claim 1 , further comprising a droplet separation region configured to increase a distance between the droplets downstream of the thermocycler and upstream of a position at which the signal is detected by the detection station.

19. The system of claim 1 , further comprising an analyzer configured to receive data from the detection station and to determine whether a nucleic acid target is present in individual droplets of the emulsion using the data.

Assignees

• Bio Rad Laboratories Inc

Inventors

• Ness Kevin D
• Masquelier Donald A
• Colston Jr Billy W
• Hindson Benjamin J

Classifications

• B01L7/525

  with physical movement of samples between temperature zones · CPC title

• B01F33/3011

  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 title

• G01N21/6486

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title

• Y02A90/10

  Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation · CPC title

• B01L2200/0689

  Sealing · CPC title