Microfluidic analysis system

• US 7,622,076 B2
• Filed: 03/03/2006
• Issued: 11/24/2009
• Est. Priority Date: 09/05/2003
• Status: Active Grant

• Current Assignee: Stokes Bio Ltd. (Perspecta, Inc.)
• Original Assignee: Stokes Bio Ltd. (Perspecta, Inc.)
• Inventors: Davies, Mark, Dalton, Tara
• Primary Examiner(s): Griffin; Walter D
• Assistant Examiner(s): Ramdhanie; Bobby
• Application Number:
  US11/366,524
• Publication Number:
  US 20060205062A1
• Time in Patent Office: 1,362 Days
• Field of Search: 422/57
• US Class Current: 422/504
• CPC Class Codes: B01F 25/10 Mixing by creating a vortex flow, e.g. by tangential introduction of flow components
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/141 Preventing contamination, tampering
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• 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 3/502776 specially adapted for focusing or laminating flows
• 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 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2035/00514 Stationary mixing elements
• CPC Class Codes: G01N 35/08 using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G01N 35/1095 for supplying the samples to flow-through analysers for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00

Abstract

A microfluidic analysis system (1) performs polymerase chain reaction (PCR) analysis on a bio sample. In a centrifuge (6) the sample is separated into DNA and RNA constituents. The vortex is created by opposing flow of a silicon oil primary carrier fluid effecting circulation by viscous drag. The bio sample exits the centrifuge enveloped in the primary carrier fluid. This is pumped by a flow controller (7) to a thermal stage (9). The thermal stage (9) has a number of microfluidic devices (70) each having thermal zones (71, 72, 73) in which the bio sample is heated or cooled by heat conduction to/from a thermal carrier fluid and the primary carrier fluid. Thus, the carrier fluids envelope the sample, control its flowrate, and control its temperature without need for moving parts at the micro scale.

Claims

1. A biological sample analysis system comprising:

a carrier fluid supply;

a sample supply;

a sample preparation stage for providing a flow of sample droplets enveloped in an immiscible primary carrier fluid;

at least one channel for continuous flow of the carrier fluid which carries the sample droplets;

at least one analysis stage for performing analysis of the sample droplets while continuously enveloped within the primary carrier fluid without the sample droplets contacting a solid surface of the channel; and

a controller for controlling the system.

2. The biological sample analysis system as claimed in claim 1, wherein there are a plurality of analysis stages, and said analysis stages comprise a thermal cycling stage and an optical detection stage for performance of a polymerase chain reaction.

3. The biological sample analysis system as claimed in claim 1, wherein there are a plurality of analysis stages, and said analysis stages comprise a thermal cycling stage and an optical detection stage for performance of a polymerase chain reaction;

and wherein the sample preparation stage comprises a centrifuge for separation of samples from an input fluid and for introduction of the samples to the primary carrier fluid.

6. The biological sample analysis system as claimed in claim 3, wherein the controller directs separation in the centrifuge either radially or axially due to gravity according to nature of the input fluid such as blood containing the sample.

8. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid.

9. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the microfluidic thermal device thermal zone further comprises separate sample and thermal outlets positioned to allow flow of thermal carrier fluid into and out of contact with the primary carrier fluid.

10. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

wherein the microfluidic thermal device thermal zone further comprises separate sample and thermal outlets positioned to allow flow of thermal carrier fluid into and out of contact with the primary carrier fluid; and

wherein there is at least one pair of opposed thermal carrier inlet/outlet pairs on opposed sides of a sample channel.

11. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the thermal cycling stage comprises a plurality of thermal zones.

12. The biological sample analysis system as claimed in claim 11, wherein the microfluidic thermal device comprises a plurality of thermal zones in series.

13. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the thermal cycling stage comprises a plurality of microfluidic thermal devices in series.

14. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the microfluidic thermal device comprises a closed sample channel for re-circulation of sample with successive heating or cooling in successive thermal zones.

15. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the controller directs flow of the thermal and primary carrier fluids to control flowrate of sample by enveloping within the primary carrier fluid and by viscous drag between the thermal carrier fluid and the primary carrier fluid.

16. The biological sample analysis system as claimed in claim 1, wherein the primary carrier fluid is biologically non-reactive.

17. The biological sample analysis system as claimed in claim 16, wherein the primary carrier fluid is a silicone oil.

18. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the thermal carrier fluid is biologically non-reactive.

19. The biological sample analysis system as claimed in claim 18, wherein the thermal carrier fluid is a silicone oil.

20. The biological sample analysis system as claimed in claim 2, wherein the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid;

and

wherein the temperatures and flowrates of the carrier fluids are controlled to achieve a temperature ramping gradient of 17°

C./sec to 25°

C./sec.

4. A biological sample analysis system comprising:

a carrier fluid supply;

a sample supply;

a sample preparation stage for providing a flow of sample droplets enveloped in an immiscible primary carrier fluid;

at least one channel for continuous flow of the carrier fluid which carries the sample droplets;

at least one analysis stage for performing analysis of the sample in the continuous flow while enveloped within the primary carrier fluid without the sample contacting a solid surface of the channel; and

a controller for controlling the system,

wherein there are a plurality of analysis stages, and said analysis stages comprise a thermal cycling stage and an optical detection stage for performance of a polymerase chain reaction;

wherein the sample preparation stage comprises a centrifuge for separation of samples from an input fluid and for introduction of the samples to the primary carrier fluid; and

wherein the centrifuge comprises a pair of opposed primary carrier fluid channels on either side of a vortex chamber, whereby flow of primary carrier fluid in said channels causes centrifuging of sample in the vortex chamber and flow of sample from the chamber into said channels.

5. The biological sample analysis system as claimed in claim 4, wherein contact between the sample and the vortex chamber surface is avoided by wrapping the sample in an initial carrier fluid within the chamber.

7. The biological sample analysis system as claimed in claim 4, wherein the primary carrier fluid velocity is in the range of 1 m/s to 20 m/s.

21. A biological sample analysis system comprising:

at least one channel for continuous flow of sample droplets enveloped in an immiscible carrier fluid, wherein the droplets do not contact a surface of the channel; and

an analysis stage for analyzing the sample droplets while the droplets are flowing through the channel.