APPARATUS AND METHODS FOR CONDUCTING ASSAYS AND HIGH THROUGHPUT SCREENING

• US 20080274493A1
• Filed: 05/27/2008
• Published: 11/06/2008
• Est. Priority Date: 11/16/2000
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Unger, Marc A., Chou, Hou-Pu, Scherer, Axel, Quake, Stephen R., Thorsen, Todd A.
• Granted Patent: US 7,887,753 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: B01D 2313/08 Flow guidance means within the module or the apparatus
• CPC Class Codes: B01D 2313/60 Specific sensors or sensor arrangements
• CPC Class Codes: B01D 2313/90 Additional auxiliary systems integrated with the module or apparatus
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 61/28 Apparatus therefor
• CPC Class Codes: B01D 63/081 Manufacturing thereof
• CPC Class Codes: B01D 63/088 Microfluidic devices comprising semi-permeable flat membranes
• CPC Class Codes: B01F 25/433 Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• CPC Class Codes: B01F 27/00 Mixers with rotary stirring devices in fixed receptacles magnetic mixers B01F33/45; Kneaders
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/81 Combinations of similar mixers, e.g. with rotary stirring devices in two or more receptacles
• CPC Class Codes: B01F 33/813 mixing simultaneously in two or more mixing receptacles
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• 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 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 : Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 : comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 : Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0877 : Flow chambers
• CPC Class Codes: B01L 2300/0887 : Laminated structure
• CPC Class Codes: B01L 2300/123 : Flexible; Elastomeric
• CPC Class Codes: B01L 2300/1822 : using Peltier elements
• CPC Class Codes: B01L 2300/1827 : using resistive heater
• CPC Class Codes: B01L 2400/0415 : electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0633 : with moving parts
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/082 : Active control of flow resistance, e.g. flow controllers
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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: C12M 23/16 : Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 41/46 : of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: F04B 43/043 : Micropumps
• CPC Class Codes: F04B 43/14 : having plate-like flexible members
• CPC Class Codes: F16K 2099/0074 : using photolithography, e.g. etching
• CPC Class Codes: F16K 2099/008 : Multi-layer fabrications
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0009 : the valve element held by multiple arms
• CPC Class Codes: F16K 99/0015 : Diaphragm or membrane valves
• CPC Class Codes: F16K 99/0046 : using magnets
• CPC Class Codes: F16K 99/0048 : using piezoelectric means
• CPC Class Codes: F16K 99/0051 : using electrostatic means
• CPC Class Codes: F16K 99/0059 : actuated by a pilot fluid
• CPC Class Codes: G01N 15/06 : Investigating concentration of particle suspensions by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10
• CPC Class Codes: G01N 27/44743 : Introducing samples
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/00 : Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
• CPC Class Codes: G01N 33/5005 : involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5008 : for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/56966 : Animal cells
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The present invention provides microfluidic devices and methods for using the same. In particular, microfluidic devices of the present invention are useful in conducting a variety of assays and high throughput screening. Microfluidic devices of the present invention include elastomeric components and comprise a main flow channel; a plurality of branch flow channels; a plurality of control channels; and a plurality of valves. Preferably, each of the valves comprises one of the control channels and an elastomeric segment that is deflectable into or retractable from the main or branch flow channel upon which the valve operates in response to an actuation force applied to the control channel.

Claims

1. A microfluidic device for conducting assays, comprising:

(a) a main flow channel adapted to allow the flow of a solution therethrough;

(b) a plurality of branch flow channels, wherein each branch flow channel is in fluid communication with the main flow channel and comprises a detection region;

(c) a plurality of control channels; and

(d) a plurality of valves operatively disposed with respect to the main flow channel and/or the plurality of branch flow channels to regulate flow of the solution through the main and branch flow channels, wherein each of the valves comprises one of the control channels and an elastomeric segment that is deflectable into or retractable from the main or branch flow channel upon which the valve operates in response to an actuation force applied to the control channel, the elastomeric segment when positioned in the flow channel restricting solution flow therethrough.

2. The microfluidic device of claim 1, wherein the control channel of each valve is separated from the flow channel in which the valve operates by the elastomeric segment.

3. The microfluidic device of claim 1 further comprising a solution inlet in fluid communication with the main flow channel for introduction of the solution.

4. The microfluidic device of claim 3 further comprising a second solution inlet for each branch flow channel in fluid communication therewith for introduction of a second solution.

5. The microfluidic device of claim 3 further comprising an auxiliary inlet in fluid communication with the main flow channel.

6. The microfluidic device of claim 5 further comprising an inlet flow channel in fluid communication with the main flow channel, and wherein the solution inlet and the auxiliary inlet are in fluid communication with the inlet flow channel.

7. The microfluidic device of claim 1 further comprising a detector operatively disposed with respect to at least one of the detection regions to detect an event or agent within the detection region.

8. The microfluidic device of claim 7, wherein the detector detects an optical signal from the detection region.

9. The microfluidic device of claim 8, wherein the detector can detect a fluorescence emission, fluorescence polarization or fluorescence resonance energy transfer.

10. The microfluidic device of claim 9, wherein the detector can perform time-resolved fluorescence measurements of fluorescence correlation spectroscopy.

11. The microfluidic device of claim 8, wherein the detector is an optical microscope, a confocal microscope or a laser scanning confocal microscope.

12. The microfluidic device of claim 7, wherein the detector is a non-optical sensor selected from the group consisting of a temperature sensor, a conductivity sensor, a potentiometric sensor and an amperometric sensor.

13. The microfluidic device of claim 1, wherein each branch flow channel is adapted to allow the flow of a second solution therethrough.

14. The microfluidic device of claim 1, wherein the main flow channel is one of a plurality of main flow channels, each adapted to allow the flow of a first solution therethrough and in fluid communication with each of the branch flow channels.

15. The microfluidic device of claim 1, wherein each of the branch flow channels intersect with the main flow channel at a different intersection, and further comprising a chamber located at each of the intersections, the chamber being adapted to collect solution therein.

16. The microfluidic device of claim 1 further comprising a plurality of pumps, and wherein each pump is operatively disposed with respect to one of the branch flow channels such that solution flow through each of the branch flow channels can be regulated by one of the pumps.

17. The microfluidic device of claim 16, wherein each of the plurality of pumps comprises at least three control channels each formed within an elastomeric material and separated from the branch flow channel by a section of an elastomeric membrane, the membrane being deflectable into the branch flow channel in response to an actuation force.

18. The microfluidic device of claim 1 further comprising a plurality of mixers, wherein each mixer is operatively disposed with respect to one of the branch flow channels and is adapted to receive and mix different solutions flowing through one of the branch flow channels.

19. The microfluidic device of claim 18 further comprising a temperature controller operatively disposed to regulate temperature within the mixer.

20. The microfluidic device of claim 18, wherein the mixer comprises

(i) an inlet section and an outlet section;

(ii) a looped flow channel in fluid communication with the inlet and outlet section; and

(iii) a pump operatively connected to the looped flow channel, wherein the pump comprises a plurality of control channels, each separated from the looped flow channel by a segment of an elastomeric membrane that is deflectable into the looped flow channel in response to an actuation force, and wherein the mixer is in fluid communication with at least one of the flow channels such that solution from the flow channel can enter via the inlet section and reenter the flow channel via the outlet section, and solution within the mixer can be cycled through the looped flow channel by the pump.

21. The microfluidic device of claim 20, wherein the detection region comprises the mixer.

22. The microfluidic device of claim 1 further comprising a temperature controller.

23. The microfluidic device of claim 22 further comprising a plurality of temperature controllers, wherein each temperature controller is operatively disposed with respect to one of the branch channels.

24. The microfluidic device of claim 22, wherein the temperature controller is disposed to regulate temperature within the detection region.

25. The microfluidic device of claim 1 further comprising a plurality of separation units, wherein each separation unit is operatively disposed with respect to one of the branch flow channels.

26. The microfluidic device of claim 25, wherein each separation unit comprises a semi-penneable membrane that separates one of the branch flow channels and a collection channel, the membrane allowing passage of certain agents in a solution flowing through the branch flow channel to pass through the semi-penneable membrane into the collection flow channel.

27. The microfluidic device of claim 25, wherein the separation unit comprises a separation material that separates molecules on the basis of affinity, size, charge or mobility.

28. The microfluidic device of claim 1 further comprising:

(e) an inlet in fluid communication with the main flow channel for introduction of the solution and an inlet for each branch flow channel in fluid communication therewith for introduction of a second solution;

(f) a plurality of chambers positioned at points where the main flow channel and the branch flow channels intersect, the chambers being adapted to collect solution therein;

(g) a plurality of pumps, each pump being operatively disposed with respect to one of the branch flow channels such that solution flow through each of the branch flow channels can be regulated by one of the pumps; and

(h) a plurality of mixers, each mixer being operatively disposed with respect to one of the branch flow channels and being adapted to receive and mix different solutions flowing through one of the branch flow channels.

29. The microfluidic device of claim 1 further comprising a pair of storage valves operatively disposed with respect to each of the branch flow channels, the pair of storage valves being disposed with respect to one another such that when the elastomeric segment of each storage valve of the pair extends into the branch flow channel a holding space is formed between the segments in which the solution can be retained.

30. The microfluidic device of claim 29 further comprising a plurality of control valves operatively disposed with respect to the main flow channel, and wherein

the control valves are positioned along the main flow channel such that by selectively actuating the control valves solution can be controllably introduced into the branch flow channels;

and

the control valves each comprise one of the control channels and an elastomeric segment that is deflectable into or retractable from the main flow channel in response to an actuation force applied to the control channel.

31. The microfluidic device of claim 30, wherein the elastomeric segments of at least one pair of storage valves each comprise one or more protrusions, the protrusions adapted to allow for the solution to flow through the holding space once the elastomeric segments are deflected into the branch flow channel while capable of retaining a particle that is present in the solution within the holding space of the at least one pair of storage valves.

32. The microfluidic device of claim 30, wherein the branch flow channel is formed within an elastomeric material and a segment of the branch flow channel opposite the elastomeric segment of each storage valve of at least one pair of storage valves comprises one or more elastomeric protrusions, the protrusions adapted to allow for solution to flow through the holding space once the elastomeric segment is deflected into the branch flow channel while retaining a particle that is present in the solution within the holding space of the at least one pair of valves.

33. The microfluidic device of claim 30, wherein

the branch flow channels are formed within an elastomeric material;

the elastomeric segments of at least one pair of storage valves comprise one or more protrusions;

a segment of the branch flow channel opposite the elastomeric segments of the at least one storage valve pair comprises one or more protrusions, and wherein

the protrusions of the elastomeric segment and the protrusions of the branch flow channel are adapted to allow for solution to flow through the holding space of the at least one valve pair once the elastomeric segments are deflected into the branch flow channel while retaining a particle that is present in the solution within the holding space.

34. The microfluidic device of claim 29 further comprising a plurality of chambers positioned at points where an inlet flow channel and the branch flow channels intersect, the chambers adapted for storing solution.

35. The microfluidic device of claim 29 further comprising a plurality of branch control valves, wherein a branch control valve is operatively disposed with respect to each of the branch flow channels and each branch control valve comprises an elastomeric segment that separates one of the branch flow channels and one of the control channels and that is deflectable into or retractable from the branch flow channel upon which the valve operates in response to an actuation force applied to the control channel.

36. The microfluidic device of claim 29, wherein each branch flow channel is in communication with a pump, the pump comprising at least three of the control channels, each formed within an elastomeric material and separated from one of the branch flow channels by an elastomeric membrane, the membrane being deflectable into the branch flow channel in response to an actuation force.

37. The microfluidic device of claim 29, wherein each branch flow channel comprises a particle enrichment section that selectively retains particles of interest.

38. The microfluidic device of claim 37, wherein the enrichment section contains a ligand that is capable of specifically binding to particle (s) that are present in the solution.

39. The microfluidic device of claim 38, wherein the detection region includes the holding space and a detector is disposed to detect particle (s) within the holding spaces within the plurality of branch channels.

40. The microfluidic device of claim 29, wherein the detection region includes the hold space.

41. The microfluidic device of claim 29, wherein the detection region is located at a section of the branch flow channel other than the holding space.

42. A microfluidic device for conducting cellular assays, comprising:

(a) a main flow channel;

(b) an input adapted to receive a first solution and in fluid communication with the main flow channel, whereby solution introduced into the input can flow into the main flow channel;

(d) a plurality of control channels;

(e) a plurality of chambers positioned along the main flow channel and in fluid communication therewith, such that solution flowing through the main flow channel can be stored in the chambers;

(f) a plurality of branch flow channels, wherein each branch flow channel is in fluid communication with a branch inlet, each branch inlet adapted to receive a second solution and in fluid communication with one of the chambers, different branch flow channels being in fluid communication with different chambers;

(g) a plurality of storage valves, wherein a pair of storage valves are operatively disposed with respect to each of the branch flow channels, and wherein each of the storage valves comprises an elastomeric segment that separates one of the branch flow channels and one of the control channels and that is deflectable into or retractable from the flow channel upon which the valve operates in response to an actuation force applied to the control channel, the storage valves of a pair being disposed with respect to one another such that when the elastomeric segment of each storage valve of the pair extends into the branch flow channel a holding space is formed between the segments in which one or more cells can be retained; and

(h) one or more pumps to transport the first and/or second solution through the branch flow channels.

43. The microfluidic device of claim 42, wherein each pump comprises at least three control channels, each formed within an elastomeric material and separated from one of the branch flow channels by a section of an elastomeric membrane, the membrane being deflectable into the branch flow channel in response to an actuation force.

44. A method of conducting a cellular analysis, comprising

(a) providing a microfluidic device comprising

(i) a flow channel comprising a detection region;

(ii) a plurality of control channels; and

(iii) a pair of storage valves operatively disposed with respect to the flow channel, wherein each storage valve comprises one of the control channels, which control channel is separated from the flow channel by an elastomeric membrane, the elastomeric membrane being deflectable into the flow channel in response to an actuation force applied to the control channel, the two control channels of the storage valves being disposed relative to one another such that deflection of the elastomeric membrane of the respective control channels into the flow channel forms a holding space within the flow channel;

(b) introducing a sample containing one or more cells into the flow channel;

(c) actuating the storage valves to hold at least one of the cells within the holding space; and

(d) performing an assay by contacting the cells within the holding space with a solution containing one or more assay agents.