Microfluidic devices

• US 8,476,063 B2
• Filed: 06/15/2010
• Issued: 07/02/2013
• Est. Priority Date: 09/15/2004
• Status: Active Grant

• Current Assignee: IntegenX Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: IntegenX Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Jovanovich, Stevan Bogdan, Blaga, Iuliu Ioan
• Primary Examiner(s): Bhat, Narayan
• Application Number:
  US12/815,685
• Publication Number:
  US 20110076735A1
• Time in Patent Office: 1,113 Days
• Field of Search: 435/6.1, 435/91.2, 435/173.9, 435/283.1, 435/287.2, 435/288.6, 435/302.1, 204/451, 536/24.33
• US Class Current: 435/287.2
• 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: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• 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 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0803 Disc shape
• 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 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 : electrophoretic flow
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• 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 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: 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: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12N 15/1003 : Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12N 15/1013 : by using magnetic beads
• CPC Class Codes: C12Q 1/24 : Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6869 : Methods for sequencing
• CPC Class Codes: C12Q 1/70 : involving virus or bacteriophage immunoassay for viruses G01N33/56983
• CPC Class Codes: G01N 2035/00514 : Stationary mixing elements
• CPC Class Codes: Y10T 436/163333 : Organic [e.g., chemical warfare agents, insecticides, etc.]
• CPC Class Codes: Y10T 436/25875 : Gaseous sample or with change of physical state

Abstract

Methods and devices for the interfacing of microchips to various types of modules are disclosed. The technology disclosed can be used as sample preparation and analysis systems for various applications, such as DNA sequencing and genotyping, proteomics, pathogen detection, diagnostics and biodefense.

Claims

1. An instrument configured to perform variable number tandem repeat (VNTR) analysis on a cell-containing sample, comprising:

(a) an integrated set of modules comprising;

(i) a capturing and concentrating off-chip macroscale module comprising;

(1) a reservoir dimensioned to receive a liquid having an initial sample volume of a milliliter to a liter,

(2) magnetic beads contained in the macroscale module configured for capturing and concentrating a DNA analyte from said initial sample volume; and

(3) a reservoir containing lysis buffer;

(ii) a bioprocessor module comprising;

(1) a microfluidic chip comprising a microfluidic channel in fluidic communication with the macroscale module and configured to receive the magnetic beads from said macroscale module,

(2) PCR primers for amplifying variable number tandem repeats, and

(3) a reaction chamber in fluid communication with the microfluidic channel, wherein the reaction chamber is configured to receive the DNA analyte and for performing a nucleic acid amplification reaction on the DNA analyte to produce a processed sample; and

(iii) an analysis module fluidically connected to said reaction chamber and comprising a capillary array electrophoresis device;

(b) a magnet configured to immobilize the magnetic beads with the captured DNA in the macroscale module and to release the magnetic beads with the captured DNA analyte into a reduced sample volume as compared to the initial sample volume;

(c) a pressure source configured for moving the magnetic beads from the macroscale module into the microfluidic channel, wherein the magnetic beads with captured DNA analyte move into the microfluidic chip in said reduced sample volume;

(d) a thermal cycling temperature control element for performing thermal cycling in the reaction chamber; and

(e) a computer comprising software programmed to control the pressure source and the thermal cycling control element.

2. The instrument of claim 1, wherein said magnetic beads are carboxylated.

3. The instrument of claim 1, wherein said microfluidic chip comprises at least one pneumatically operated diaphragm valve.

4. The instrument of claim 1, wherein the microfluidic chip comprises a fluidic layer, a deformable membrane layer and a pneumatic layer, said fluidic layer comprising the microfluidic channel, and said at least one pneumatically operated diaphragm valve comprising the deformable membrane layer.

5. The instrument of claim 4 wherein said deformable membrane layer comprises polydimethylsiloxane.

6. The instrument of claim 1, wherein said reagents for amplifying VNTRs comprise reagents for performing PCR.

7. The instrument of claim 1, wherein said microfluidic microchip is disposable or reusable.

8. The instrument of claim 1, wherein said software sequentially operates pneumatic solenoids so as to create a pumping action that drives fluid in the microfluidic channel.

9. The instrument of claim 1, wherein said capillary electrophoresis array is on-chip.

10. The instrument of claim 1, wherein said capillary electrophoresis array is off-chip.

11. The instrument of claim 1, wherein the capillary electrophoresis array is arranged radially.

12. The instrument of claim 1, further comprising a waste reservoir.

13. A method for performing variable number tandem repeat (VNTR) analysis on a cell-containing sample, comprising the steps of:

a) concentrating a DNA analyte in a macroscale module by;

(i) providing a sample comprising cells comprising a DNA analyte in an initial volume into an off-chip macroscale module configured to hold a volume of a milliliter to a liter;

(ii) lysing cells in the sample to release DNA from said initial volume;

(iii) capturing the released DNA analyte onto a plurality of magnetic beads in the macroscale module;

(iv) immobilizing the magnetic beads with the captured DNA analyte in the macroscale module with a magnet;

(v) washing the magnetic beads with the captured DNA analyte; and

(vi) releasing the washed beads into a reduced volume compared with said initial volume to concentrate the DNA analyte;

b) introducing, with pressure from a pressure source, the released magnetic beads with the captured DNA analyte in the reduced volume into a bioprocessor module that is fluidically integrated with the macroscale module, wherein introducing comprises passing the magnetic beads with the captured DNA analyte from the macroscale module through a microfluidic channel in a microfluidic chip and into a reaction chamber in the bioprocessor module wherein the microfluidic channel is in fluidic communication with the macroscale module and with the reaction chamber;

c) amplifying variable number tandem repeats (VNTRs) in the DNA analyte in the reaction chamber using PCR primers; and

d) analyzing the amplified DNA analyte by capillary electrophoresis in an analysis module fluidically integrated with the bioprocessor module.

14. The method of claim 13 wherein the initial volume comprising the analyte in the macroscale module is one milliliter to one liter.

15. The method of claim 13 further comprising eluting the analytes from the magnetic beads in the bioprocessor module.

16. The method of claim 13 wherein the introducing comprises pumping the magnetic beads into the microfluidic chip using an on-chip diaphragm pump comprising a series of three diaphragm valves.

17. The method of claim 13 further comprising moving a lysis buffer into a chamber in the macroscale module that holds the cells.

18. The method of claim 13 wherein amplifying comprises thermal cycling.

19. The method of claim 13, comprising performing capillary electrophoresis off-chip.

20. The method of claim 13 performed on a plurality of samples.