Integrated system for processing microfluidic samples, and method of using the same

• US 9,040,288 B2
• Filed: 03/26/2007
• Issued: 05/26/2015
• Est. Priority Date: 03/24/2006
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Handique, Kalyan, Brahmasandra, Sundaresh N., Ganesan, Karthik, Wu, Betty, Phadke, Nikhil, Parunak, Gene, Williams, Jeff
• Primary Examiner(s): Marcheschi, Michael
• Assistant Examiner(s): Doe, Shanta G
• Application Number:
  US11/728,964
• Publication Number:
  US 20070292941A1
• Time in Patent Office: 2,983 Days
• Field of Search: 435/6, 4352851-2852, 4352871-2873, 4352885-2887, 435/303.1
• US Class Current: 435/288.5
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• 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 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0611 duck bill valves
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 2400/0683 : mechanically breaking a wall or membrane within a channel or chamber
• 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 3/50273 : 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 7/52 : 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: 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 99/003 : Valves for single use only
• CPC Class Codes: F16K 99/0032 : using phase transition or influencing viscosity
• CPC Class Codes: F16K 99/0044 : using thermo-electric means
• CPC Class Codes: F16K 99/0061 : actuated by an expanding gas or liquid volume
• CPC Class Codes: G01N 2035/00881 : network configurations
• CPC Class Codes: Y02A 90/10 : Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

This patent application describes an integrated apparatus for processing polynucleotide-containing samples, and for providing a diagnostic result thereon. The apparatus is configured to receive a microfluidic cartridge that contains reagents and a network for processing a sample. Also described are methods of using the apparatus.

Claims

1. An apparatus, comprising:

a receiving bay configured to receive an insertable multi-lane microfluidic cartridge comprising a plurality of PCR reaction zones, each reaction zone configured to accept a polynucleotide-containing sample, each lane comprising one of the plurality of PCR reaction zones;

a processor;

a memory in communication with the processor;

a plurality of heater sets fixed in position in the receiving bay, each heater set configured to thermally couple to one of the plurality of PCR reaction zones in the multi-lane cartridge, each heater set comprising a plurality of heat sources, the memory comprising instructions that direct the processor to independently control the temperature of the plurality of heat sources in each heater set to cyclically heat one of the plurality of PCR reaction zones in a series of heating phases without moving the plurality of heat sources relative to each other, the memory further comprising instructions that direct the processor to control the temperature of the plurality of heat sources to maintain a substantially uniform temperature throughout one of the plurality of PCR reaction zones during each heating phase without moving the plurality of heat sources relative to each other, the memory further comprising instructions that direct the processor to perform independent PCR reactions on polynucleotide-containing samples in the multi-lane cartridge; and

a detector configured to detect the presence of one or more amplified polynucleotides on the multi-lane microfluidic cartridge.

2. The apparatus of claim 1, further comprising a registration member that is complementary to the multi-lane microfluidic cartridge, whereby the receiving bay receives the multi-lane microfluidic cartridge in a single orientation.

3. The apparatus of claim 1, further comprising a sensor coupled to the processor, the sensor configured to sense whether the multi-lane microfluidic cartridge is received.

4. The apparatus of claim 1, wherein the detector is an optical detector.

5. The apparatus of claim 4, wherein the optical detector comprises a light source configured to emit light in an absorption band of a fluorescent dye, and wherein the optical detector further comprises a light detector configured to detect light in an emission band of the fluorescent dye, wherein the fluorescent dye corresponds to a fluorescent polynucleotide probe or a fragment thereof.

6. The apparatus of claim 4, wherein the optical detector is configured to independently detect a plurality of fluorescent dyes at a plurality of different locations of the multi-lane microfluidic cartridge, wherein each fluorescent dye corresponds to a fluorescent polynucleotide probe or a fragment thereof.

7. The apparatus of claim 1, wherein the processor is programmable to operate the detector to detect a polynucleotide or a probe thereof in the multi-lane microfluidic cartridge.

8. The apparatus of claim 1, wherein each of the plurality of heater sets comprises at least two contact heat sources.

9. The apparatus of claim 8, wherein at least one of the at least two contact heat sources is selected from a group consisting of:

a resistive heater, a radiator, a fluidic heat exchanger, and a Peltier device.

10. The apparatus of claim 8, wherein the at least two contact heat sources are configured to be in direct physical contact with the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is inserted into the receiving bay.

11. The apparatus of claim 1, wherein at least one of the plurality of heater sets has a heating area of between about 1 mm² and about 225 mm².

12. The apparatus of claim 11, wherein the heating area is between about 1 mm² and about 100 mm².

13. The apparatus of claim 8, further comprising a compliant layer at the at least two contact heat sources, wherein the compliant layer is configured to thermally couple the at least two contact heat sources with one or more selected regions of the multi-lane microfluidic cartridge.

14. The apparatus of claim 13, wherein the compliant layer has a thickness of between about 0.05 and about 2 millimeters and a Shore hardness of between about 25 and about 100.

15. The apparatus of claim 1, wherein at least one of the plurality of heat sources is a resistive heater.

16. The apparatus of claim 1, further comprising a heating stage configured to be removable from the apparatus wherein at least one of the plurality of heat sources is located in the heating stage.

17. The apparatus of claim 1, further comprising a lid at the receiving bay, the lid being operable to at least partially exclude ambient light from the receiving bay.

18. The apparatus of claim 17, wherein the lid is a sliding lid.

19. The apparatus of claim 1, wherein the plurality of heater sets comprise a plurality of resistive heaters.

20. The apparatus of claim 19, wherein the plurality of resistive heaters in each heater set align with one PCR reaction zone in one lane of the multi-lane microfluidic cartridge when the multi-lane microfluidic cartridge is inserted into the receiving bay.

21. The apparatus of claim 1, wherein the apparatus is configured to direct, under the control of the processor, electrical signals to each of the plurality of heat sources in each heater set independently.

22. The apparatus of claim 1, further comprising one or more force members configured to apply force to at least a portion of the multi-lane microfluidic cartridge.

23. The apparatus of claim 22, wherein the one or more force members are configured to operate a mechanical member at the multi-lane microfluidic cartridge.

24. The apparatus of claim 23, wherein the mechanical member is a pierceable reservoir.

25. The apparatus of claim 22, wherein the one or more force members are configured to apply force to a plurality of locations in the multi-lane microfluidic cartridge.

26. The apparatus of claim 22, wherein the force applied by the one or more force members results in an average pressure at an interface between a portion of the receiving bay and a portion of the multi-lane microfluidic cartridge of between about 5 kilopascals and about 50 kilopascals.

27. The apparatus of claim 26, wherein the average pressure is at least about 14 kilopascals.

28. The apparatus of claim 22, wherein at least one of the one or more force members is manually operated.

29. The apparatus of claim 22, wherein at least one of the one or more force members is mechanically coupled to a lid at the receiving bay, whereby operation of the lid operates the force member.

30. The apparatus of claim 1, further comprising at least one input device coupled to the processor, the at least one input device being selected from a group consisting of a keyboard, a touch-sensitive surface, a microphone, a track-pad, a retinal scanner, a fingerprint reader, and a mouse.

31. The apparatus of claim 1, further comprising a data storage medium configured to receive data from one or more of a processor, an input device, or a communication interface, the data storage medium being selected from a group consisting of:

a hard disk drive, an optical disk drive, a flash-card, and a CD-Rom.

32. The apparatus of claim 1, further comprising a communication interface coupled to the processor, the communication interface being selected from a group consisting of:

a serial connection, a parallel connection, a wireless network connection, and a wired network connection.

33. The apparatus of claim 1, further comprising at least one sample identifier coupled to the processor, the at least one sample identifier being selected from a group consisting of:

an optical character reader, a bar code reader, and a radio frequency tag reader.

34. The apparatus of claim 33, wherein the at least one sample identifier is a handheld bar code reader.

35. The apparatus of claim 1, further comprising at least one output device coupled to the processor, the at least one output device being selected from a group consisting of:

a visual display, a printer, and a speaker.

36. The apparatus of claim 1, wherein the detector is configured to detect the presence of one or more amplified polynucleotides that are amplified by a method selected from a group consisting of:

polymerase chain reaction;

TMA;

SDA;

NASBA;

LCR; and

Rolling-Cycle Amplifications.

37. A system comprising the apparatus of claim 1 and a multi-lane microfluidic cartridge inserted into the receiving bay of the apparatus, wherein the multi-lane microfluidic cartridge comprises:

a sample inlet for receiving a quantity of biological sample containing one or more polynucleotides; and

a port permitting detection of one or more amplified polynucleotides.

38. A system comprising the apparatus of claim 1 and a multi-lane microfluidic cartridge inserted into the receiving bay of the apparatus, wherein the multi-lane microfluidic cartridge comprises:

one or more reagents configured to;

lyse cells in the sample;

prepare the polynucleotides for amplification; and

amplify the polynucleotides.

39. The system of claim 38, wherein the one or more reagents are held in one or more reagent retention vessels.

40. The system of claim 38, wherein the one or more reagents are held in a microfluidic chamber or a channel.

41. A system comprising the apparatus of claim 1 and a multi-lane microfluidic cartridge inserted into the receiving bay of the apparatus, wherein the multi-lane microfluidic cartridge comprises:

one or more microfluidic components configured to act on microfluidic volumes of polynucleotide containing sample before, during and after amplification of one or more polynucleotides from the sample, the one or more microfluidic components being selected from a group consisting of;

one or more channels configured to permit passage of the microfluidic volumes;

one or more actuators configured to move the microfluidic volumes;

one or more chambers configured to hold the microfluidic volumes; and

one or more components configured to inhibit motion of the microfluidic volumes.

42. The system of claim 41, wherein the one or more components configured to inhibit motion of the microfluidic volumes comprise:

a microfluidic valve configured to transform from an open to a closed state or

a microfluidic gate configured to transform from a closed to an open state.

43. The system of claim 38, wherein the one or more reagents are in freeze-dried form.

44. The system of claim 37, wherein the multi-lane microfluidic cartridge further comprises a waste reservoir for storing waste generated during sample processing and waste from the biological sample.

45. The apparatus of claim 1, wherein the plurality of heat sources are configured to maintain a temperature gradient of less than 1°

C. across a width of one of the plurality of PCR reaction zones at any point along a length of one PCR reaction zone.

46. The apparatus of claim 1, wherein the plurality of heat sources comprises four heaters configured to thermally couple to four sides of one of the plurality of PCR reaction zones.

47. The apparatus of claim 1, wherein each heater set further comprises one or more temperature sensors configured to control power supplied to the plurality of heat sources, the one or more temperature sensors further configured to transmit temperature information to the processor.

48. An apparatus, comprising:

a receiving bay configured to receive an insertable multi-lane microfluidic cartridge, the receiving bay comprising a plurality of contact heating zones, each of the plurality of contact heating zones configured to be independently thermally coupled to a distinct location in one lane of the multi-lane microfluidic cartridge;

a processor;

a memory in communication with the processor and comprising instructions;

a plurality of heater sets fixed in position in the receiving bay, each heater set thermally coupled to one of the plurality of contact heating zones, each heater set comprising a plurality of heat sources, the memory comprising instructions that direct the processor to independently control the temperature of the plurality of heat sources in each heater set to cyclically heat one of the plurality of contact heating zones in a series of heating phases without moving the plurality of heat sources relative to each other, the memory further comprising instructions that direct the processor to control the temperature of the plurality of heat sources to maintain a substantially uniform temperature throughout one of the plurality of contact heating zones during each heating phase without moving the plurality of heat sources relative to each other, the memory further comprising instructions that independently operate each of the plurality of contact heating zones to carry out nucleic acid amplification on polynucleotide-containing samples in the multi-lane microfluidic cartridge; and

a detector configured to detect the presence of one or more amplified polynucleotides on the multi-lane cartridge.

49. The apparatus of claim 48, wherein one of the plurality of heater sets comprises four heaters configured to thermally couple to one of the plurality of contact heating zones.

50. The apparatus of claim 48, wherein each of the plurality of heater sets comprises at least two contact heat sources, and wherein at least one of the at least two contact heat sources is selected from a group consisting of:

a resistive heater, a radiator, a fluidic heat exchanger, and a Peltier device.

51. The apparatus of claim 50, wherein the at least two contact heat sources are configured to be in direct physical contact with one of the plurality of contact heating zones.

52. The apparatus of claim 50, wherein at least one of the plurality of heater sets has a heating area for contacting one of the plurality of contact heating zones of between about 1 mm² and about 225 mm².