Multilayered microfluidic DNA analysis system and method

• US 6,544,734 B1
• Filed: 12/09/1999
• Issued: 04/08/2003
• Est. Priority Date: 10/09/1998
• Status: Expired due to Fees

• Current Assignee: Motorola, Inc. (Motorola Solutions, Inc.)
• Original Assignee: Motorola, Inc. (Motorola Solutions, Inc.)
• Inventors: Yu, Huinan, Huang, Rong-Fong, Burdon, Jeremy W., Briscoe, Cynthia G., Grodzinski, Piotr, Marrero, Robert
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Taylor, Janell E.
• Application Number:
  US09/460,281
• Time in Patent Office: 1,216 Days
• Field of Search: 435/6, 435/91.1, 435/7.93, 536/25.32, 536/25.4
• US Class Current: 435/6.13
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00835 Comprising catalytically active material
• CPC Class Codes: B01J 2219/00873 Heat exchange
• CPC Class Codes: B01J 2219/00966 pH
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• 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 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0439 ultrasonic vibrations, vibrating piezo elements
• 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 7/00 : Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B32B 18/00 : Layered products essentially comprising ceramics, e.g. refractory products
• CPC Class Codes: B32B 2038/042 : Punching
• CPC Class Codes: B32B 2311/06 : Platinum
• CPC Class Codes: B32B 2311/08 : Silver
• CPC Class Codes: B32B 2315/02 : Ceramics
• CPC Class Codes: B81B 2201/058 : Microfluidics not provided for in B81B2201/051 - B81B2201/054
• CPC Class Codes: B81C 1/00119 : Arrangement of basic structures like cavities or channels, e.g. suitable for microfluidic systems
• CPC Class Codes: B81C 2201/019 : Bonding or gluing multiple substrate layers
• CPC Class Codes: C04B 2235/6562 : Heating rate
• CPC Class Codes: C04B 2237/341 : Silica or silicates
• CPC Class Codes: C04B 2237/343 : Alumina or aluminates
• CPC Class Codes: C04B 2237/348 : Zirconia, hafnia, zirconates or hafnates
• CPC Class Codes: C04B 2237/50 : Processing aspects relating to ceramic laminates or to the joining of ceramic articles with other articles by heating
• CPC Class Codes: C04B 2237/62 : Forming laminates or joined articles comprising holes, channels or other types of openings
• CPC Class Codes: C04B 2237/68 : Forming laminates or joining articles wherein at least one substrate contains at least two different parts of macro-size, e.g. one ceramic substrate layer containing an embedded conductor or electrode
• CPC Class Codes: C04B 2237/704 : of one or more of the ceramic layers or articles
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: F04B 19/006 : Micropumps F04B43/043 and F04B43/095 take precedence
• CPC Class Codes: F28D 2021/0029 : Heat sinks
• CPC Class Codes: F28D 9/00 : Heat-exchange apparatus having stationary plate-like or laminated conduit assemblies for both heat-exchange media, the media being in contact with different sides of a conduit wall F28F3/083, F28F3/086 take precedence
• CPC Class Codes: F28F 13/00 : Arrangements for modifying heat-transfer, e.g. increasing, decreasing F28F1/00 - F28F11/00 take precedence
• CPC Class Codes: F28F 2260/02 : having microchannels
• CPC Class Codes: G01N 2030/025 : Gas chromatography
• CPC Class Codes: G01N 2030/528 : Monolithic sorbent material
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 30/60 : Construction of the column
• CPC Class Codes: G01N 30/6004 : end pieces
• CPC Class Codes: G01N 30/6069 : with compartments or bed substructure
• CPC Class Codes: G01N 30/6095 : Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: H05K 1/0272 : Adaptations for fluid transport, e.g. channels, holes
• CPC Class Codes: H05K 1/0306 : Inorganic insulating substrates, e.g. ceramic, glass
• CPC Class Codes: H05K 3/4611 : by laminating two or more circuit boards H05K3/4652 takes precedence
• CPC Class Codes: H05K 3/4629 : laminating inorganic sheets comprising printed circuits, e.g. green ceramic sheets

Abstract

A multilayered microfluidic DNA analysis system includes a cell lysis chamber, a DNA separation chamber, a DNA amplification chamber, and a DNA detection system. The multilayered microfluidic DNA analysis system is provided as a substantially monolithic structure formed from a plurality of green-sheet layers sintered together. The substantially monolithic structure has defined therein a means for heating the DNA amplification chamber and a means for cooling the DNA amplification chamber. The means for heating and means for cooling operate to cycle the temperature of the DNA amplification chamber as required for performing a DNA amplification process, such as PCR.

Claims

1. A DNA analysis system comprising:

a sample inlet port;

a cell lysis chamber in fluid communication with said sample inlet port;

a DNA separation chamber in fluid communication with said cell lysis chamber;

a DNA amplification chamber in fluid communication with said DNA separation chamber, said DNA amplification chamber being defined by a substantially monolithic structure, said substantially monolithic structure being formed from a plurality of green-sheet layers sintered together, said green-sheet layers including particles selected from the group consisting of ceramic particles, glass particles, and glass-ceramic particles; and

a DNA detection system in fluid communication with said DNA amplification chamber.

2. The DNA analysis system of claim 1, wherein said substantially monolithic structure has defined therein a means for heating said DNA amplification chamber and a means for cooling said DNA amplification chamber.

3. The DNA analysis system of claim 2, wherein said cell lysis chamber is defined in said substantially monolithic structure.

4. The DNA analysis system of claim 3, further comprising first and second electrodes disposed on opposing surfaces of said cell lysis chamber.

5. The DNA analysis system of claim 1, wherein said DNA separation chamber is defined in said substantially monolithic structure.

6. The DNA analysis system of claim 5, further comprising an electromagnet for directing a magnetic field into said DNA separation chamber, said electromagnet being defined in said substantially monolithic structure.

7. The DNA analysis system of claim 1, wherein said DNA detection system comprises a capillary electrophoresis channel defined in said substantially monolithic structure.

8. The DNA analysis system of claim 7, further comprising a window for providing optical access to said capillary electrophoresis channel, said window being defined in said substantially monolithic structure.

9. The DNA analysis system of claim 1, wherein said DNA detection system comprises a molecular probe array, said molecular probe array being defined by a plurality of test sites formed into a substrate, each one of said test sites containing a known oligonucleotide.

10. The DNA analysis system of claim 1, further comprising a first fluid flow control system between said cell lysis chamber and said DNA separation chamber.

11. The DNA analysis system of claim 10, wherein said first fluid flow control system comprises a pair of electrodes disposed in a microfluidic channel for electroosmotic pumping.

12. The DNA analysis system of claim 1, further comprising a second fluid flow control system between said DNA separation chamber and said DNA amplification chamber.

13. The DNA analysis system of claim 12, wherein said second fluid flow control system comprises a pair of electrodes disposed in a microfluidic channel for electroosmotic pumping.

14. The DNA analysis system of claim 1, further comprising a third fluid flow control system between said DNA amplification chamber and said DNA detection system.

15. The DNA analysis system of claim 14, wherein said third fluid flow control system comprises a capillary stop disposed in a microfluidic channel, whereby said capillary stop substantially blocks the flow of fluid at low pressures but allows the flow of fluid at high pressures.

16. A method for performing DNA analysis, said method comprising:

placing in a fluidic sample containing cells in a cell lysis chamber;

lysing said cells in said cell lysis chamber to release cell contents, said cell contents including sample DNA;

passing said cell contents to a DNA separation chamber;

adsorbing said sample DNA onto a plurality of micro-beads in said DNA separation chamber;

eluting said sample DNA from said micro-beads;

passing said sample DNA to a DNA amplification chamber within a substantially monolithic structure formed from a plurality of green sheet layers sintered together;

amplifying said sample DNA in said DNA amplification chamber to produce amplified DNA; and

detecting said amplified DNA.

17. The method of claim 16, wherein said lysing step includes the step of applying an electric field to said cells.

18. The method of claim 16, wherein said detecting step includes the step of performing electrophoresis on said amplified DNA.

19. The method of claim 16, further comprising the step of conjugating said amplified DNA to a fluorophore to form conjugated DNA, wherein said detecting step includes the step of performing electrophoresis on said conjugated DNA.

20. The method of claim 18 or 19, wherein said electrophoresis is performed in an electrophoresis channel defined in said substantially monolithic structure.

21. The method of claim 19 wherein said detecting step includes the step of detecting fluorescence from said conjugated DNA.

22. The method of claim 16, further comprising the step of passing said amplified DNA to a molecular probe array, said molecular probe array being defined by a plurality of test sites formed into a substrate, each one of said test sites containing a known oligonucleotide.

23. The method of claim 21, wherein said detecting step includes the step of said amplified DNA hybridizing with said known oligonucleotide in one of said test sites.

24. The method of claim 16, further comprising the steps of conjugating said amplified DNA to a fluorophore to form conjugated DNA and passing said conjugated DNA to a molecular probe array, said molecular probe array being defined by a plurality of test sites formed into a substrate, each one of said test sites containing a known oligonucleotide.

25. The method of claim 24, wherein said detecting step includes the step of said conjugated DNA hybridizing with said known oligonucleotide in one of said test sites.

26. The method of claim 16, wherein said micro-beads are paramagnetic, further comprising the step of applying an magnetic field to entrain said micro-beads in said DNA separation chamber.

27. The method of claim 26, wherein said step of applying a magnetic field includes the step of energizing an electromagnet, said electromagnet being defined in said substantially monolithic structure.

28. The method of claim 16, wherein said green-sheet layers comprise particles selected from the group consisting of ceramic particles, glass particles, and glass-ceramic particles.

29. The method of claim 16, wherein said cell lysis chamber is in said substantially monolithic structure.

30. The method of claim 16 or 29, wherein said separation chamber is in said substantially monolithic structure.