Integrated active flux microfluidic devices and methods

• US 8,129,176 B2
• Filed: 08/05/2009
• Issued: 03/06/2012
• Est. Priority Date: 06/05/2000
• Status: Expired due to Term

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Quake, Stephen R., Chou, Hou-Pu
• Primary Examiner(s): Bullock, In Suk
• Assistant Examiner(s): Kingan, Timothy G
• Application Number:
  US12/536,129
• Publication Number:
  US 20100120018A1
• Time in Patent Office: 944 Days
• Field of Search: 436/503, 435/6, 422/103
• US Class Current: 435/287.2
• 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 33/3022 the components being formed by independent droplets which are alternated, the mixing of the components being achieved by diffusion between droplets
• CPC Class Codes: B01F 33/3039 with mixing achieved by diffusion between layers
• CPC Class Codes: B01F 35/71 Feed mechanisms with proportioning B01F35/80
• CPC Class Codes: B01F 35/712 for feeding fluids
• CPC Class Codes: B01F 35/71755 using means for feeding components in a pulsating or intermittent manner
• CPC Class Codes: B01F 35/717611 Peristaltic pumps
• CPC Class Codes: B01F 35/71805 using valves, gates, orifices or openings
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0689 Sealing
• 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 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/088 Channel loops
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0418 : electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/043 : magnetic forces
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0638 : membrane valves, flap valves
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/0694 : vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 3/02 : Burettes; Pipettes
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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 3/50273 : 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 3/502753 : characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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 1/02 : with agitation means; with heat exchange means
• CPC Class Codes: C12Q 1/6816 : characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: G01N 33/53 : Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 137/0329 : Mixing of plural fluids of diverse characteristics or conditions
• CPC Class Codes: Y10T 137/2218 : Means [e.g., valve] in control input
• CPC Class Codes: Y10T 137/2224 : Structure of body of device
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/145555 : Hetero-N
• CPC Class Codes: Y10T 436/173845 : Amine and quaternary ammonium
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The invention relates to a microfabricated device for the rapid detection of DNA, proteins or other molecules associated with a particular disease. The devices and methods of the invention can be used for the simultaneous diagnosis of multiple diseases by detecting molecules (e.g. amounts of molecules), such as polynucleotides (e.g., DNA) or proteins (e.g., antibodies), by measuring the signal of a detectable reporter associated with hybridized polynucleotides or antigen/antibody complex. In the microfabricated device according to the invention, detection of the presence of molecules (i.e., polynucleotides, proteins, or antigen/antibody complexes) are correlated to a hybridization signal from an optically-detectable (e.g. fluorescent) reporter associated with the bound molecules. These hybridization signals can be detected by any suitable means, for example optical, and can be stored for example in a computer as a representation of the presence of a particular gene. Hybridization probes can be immobilized on a substrate that forms part of or is exposed to a channel or channels of the device that form a closed loop, for circulation of sample to actively contact complementary probes. Universal chips according to the invention can be fabricated not only with DNA but also with other molecules such as RNA, proteins, peptide nucleic acid (PNA) and polyamide molecules.

Claims

1. A method for effectively mixing two or more different fluids in a microfluidic device, which method comprises:

(a) introducing the different fluids to a microfluidic device, the microfluidic device having;

(i) a loop channel communicating with at least one service channel, and

(ii) a pump associated with the loop channel, so that each of the different fluids is loaded into the loop channel; and

(b) circulating the different fluids using the pump associated with the loop channel to mix the different fluids, wherein the pump is a peristaltic pump comprising at least three cooperating microvalves acting within the loop channel.

2. A method according to claim 1, wherein the microfluidic device has at least one inlet channel and at least one outlet channel, the different fluids being loaded into the loop channel through at least one inlet channel.

3. A method according to claim 2, wherein:

the microfluidic device has two or more inlet channels; and

each of the two or more different fluids is loaded into the loop channel through a different inlet channel.

4. A method according to claim 2, wherein each inlet channel and each outlet channel is separated from the loop channel by a microvalve.

5. A method according to claim 4, wherein the microvalves separating the inlet and outlet channels from the loop channel are closed while the pump is activated.

6. A method according to claim 4, wherein the microvalves separating the inlet and outlet channels from the loop channel remain open while the pump is activated.

7. A method according to claim 1, wherein at least one of the different fluids comprises a solution of molecules.

8. A method according to claim 7, wherein the solution of molecules contains molecules selected from a group consisting of:

nucleic acid molecules, polypeptide molecules, antibody molecules.

9. A method according to claim 1, wherein at least one of the different fluids comprises a suspension of particles.

10. A method according to claim 9, wherein the suspension of particles contains particles selected from a group consisting of:

cells, virions, and microscopic beads.

11. A method according to claim 1, wherein the microfluidic device further comprises a microvalve separating the loop channel from the service channel.

12. A method for binding a sample to a target, which method comprises:

(a) introducing a fluid containing the sample to a microfluidic device, the microfluidic device having;

(i) a loop channel communicating with at least one service channel and having molecules of the target disposed therein, and

(ii) a pump associated with the loop channel, so that the fluid containing the sample is loaded into the loop channel; and

(b) circulating the fluid through the loop channel using the pump, wherein the sample binds to the target molecules disposed in the loop as the fluid circulates therethrough, and wherein the pump is a peristaltic pump comprising at least three cooperating microvalves acting within the loop channel.

13. A method according to claim 12, wherein the target molecules are selected from a group consisting of polynucleotide probes, protein probes, antibody probes, biotin and avidin.

14. A method according to claim 12, wherein the sample comprises nucleic acid molecules, protein molecules, cells or virions.

15. A method according to claim 14, wherein

the sample comprises nucleic acid molecules, and

the target comprises polynucleotide probes specific for nucleic acid molecules in the sample.

16. A method according to claim 14, wherein:

the target comprises biotin molecules, and

the sample comprises particles or molecules having a biotin specific label.

17. A method according to claim 12, wherein the loop channel resides in a layer of elastomeric material.

18. A method according to claim 12, wherein the microfluidic device further comprises a microvalve separating the loop channel from the service channel.