THERMAL CYCLING DEVICE

• US 20050186585A1
• Filed: 02/24/2005
• Published: 08/25/2005
• Est. Priority Date: 02/24/2004
• Status: Active Grant

• Current Assignee: Thermal Gradient
• Original Assignee: Thermal Gradient
• Inventors: Juncosa, Robert, Grover, Joel
• Granted Patent: US 7,618,811 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1883 using thermal insulation
• 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 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]

Abstract

A multi-layer device suitable for thermal cycling is set forth. The device is particularly suitable for performing a polymerase chain reaction (PCR). One embodiment includes a first layer (18), a second layer (20) adjacent to the first layer, and a third layer (22) adjacent to the second layer opposite the first layer. A continuous channel (54) is formed within the first layer (18), the second layer (20), and the third layer (22). The continuous channel (54) has a plurality of cycle segments, each of the cycle segments comprises a first portion (58) disposed within the first layer (18), a second portion (62) disposed within the third layer (22), and a third portion (66) disposed within the second layer (20). An integral or separate temperature source or sources (14), (16) may be provided to maintain the first layer (18) at a first temperature and the third layer (22) at a second temperature. Thermal conduction maintains the third layer (22) at a temperature between the first temperature and the second temperature.

Claims

1. An apparatus comprising:

a first layer;

a second layer proximate to the first layer; and

a continuous channel formed within the first layer and the second layer, said continuous channel having a plurality of cycle segments, each cycle segment comprising a first cycle portion disposed within the first layer and a second cycle portion disposed within the second layer.

2. An apparatus as recited in claim 1 wherein the first layer and second layer form a device for performing a polymerase chain reaction, wherein the first layer comprises an annealing and extension layer and the second layer comprises a denaturing layer.

3. An apparatus as recited in claim 1 wherein the first layer and second layer are thermally conductive.

4. An apparatus as recited in claim 1 further comprising a first insulator disposed between the first layer and second layer.

5. An apparatus as recited in claim 1 wherein the first layer is a substantially planar layer.

6. An apparatus as recited in claim 1 wherein the second layer is a substantially planar layer.

7. An apparatus as recited in claim 1 further comprising a first temperature source thermally coupled to and maintaining the first layer at a first temperature and a second temperature source thermally coupled to and maintaining the second layer at a second temperature.

8. An apparatus as recited in claim 1 wherein the first layer and the second layer are composed of the same material.

9. An apparatus as recited in claim 1 wherein the first layer and the second layer are composed of a different material.

10. An apparatus as recited in claim 1 further comprising an LED and a photodetector disposed within the continuous channel.

11. An apparatus as recited in claim 1 further comprising a third layer adjacent to the second layer opposite the first layer, wherein the continuous channel is formed within the first layer, the second layer and the third layer and wherein each cycle segment comprising the first cycle portion disposed within the first layer, the second cycle portion disposed within the third layer and a third cycle portion disposed within the second layer.

12. An apparatus as recited in claim 11 wherein the first layer, second layer, and third layer form a polymerase chain reaction device, wherein the first layer comprises a denaturing layer, the second layer comprises an extension layer and the third layer comprises an annealing layer.

13. An apparatus as recited in claim 12 wherein prior to the first cycle portion or first cycle portion and second cycle portion, said continuous channel having an extended denaturing portion.

14. An apparatus as recited in claim 11 wherein the cycle segment comprises the first portion followed by the second portion and followed by the third portion.

15. An apparatus as recited in claim 11 wherein the first layer, second layer and third layer are thermally conductive.

16. An apparatus as recited in claim 11 further comprising a first insulator disposed between the first layer and second layer.

17. An apparatus as recited in claim 16 wherein the first insulator has a thermal conductivity lower than the first layer.

18. An apparatus as recited in claim 16 wherein the first insulator comprises a first insulative layer.

19. An apparatus as recited in claim 11 further comprising a second insulator disposed between the second layer and the third layer.

20. An apparatus as recited in claim 19 wherein the second insulator has a thermal conductivity lower than the second layer.

21. An apparatus as recited in claim 19 wherein the second insulator comprises a second insulative layer.

22. An apparatus as recited in claim 11 further comprising a first temperature source maintaining the first layer at a first temperature.

23. An apparatus as recited in claim 22 wherein the first temperature source is disposed adjacent to the first layer.

24. An apparatus as recited in claim 22 wherein the first temperature source is disposed between the first layer and the second layer.

25. An apparatus as recited in claim 22 wherein the first temperature source is part of the first layer.

26. An apparatus as recited in claim 22 wherein the first temperature source contacts an edge of the first layer.

27. An apparatus as recited in claim 22 further comprising a second temperature source maintaining the third layer at a second temperature.

28. An apparatus as recited in claim 27 wherein the second layer comprises a third temperature source coupled thereto.

29. An apparatus as recited in claim 27 wherein the first temperature is greater than the second temperature.

30. An apparatus as recited in claim 29 wherein the second layer is maintained at a third temperature between the first temperature and the second temperatures.

31. An apparatus as recited in claim 11 wherein the first layer, the second layer and the third layer are composed of the same material.

32. An apparatus as recited in claim 11 wherein at least two of the first layer, the second layer and the third layer are composed of a different material.

33. An apparatus as recited in claim 11 wherein the first layer is a substantially planar layer, the second layer is a substantially planar layer, and the third layer is a substantially planar layer.

40. A device as recited in claim 32 further comprising a cover layer disposed on the first thermally conductive layer.

41. A device as recited in claim 40 wherein a cover layer is transparent.

34. A device for performing a reaction comprising:

a first heating means;

a first thermally conductive layer thermally coupled to the first heating means;

a first insulating layer proximate the first thermally conductive layer;

a second thermally conductive layer directly adjacent to the first insulating layer;

a second insulating layer directly adjacent to the second thermally conductive layer opposite the first insulating layer;

a third thermally conductive layer proximate to the second layer opposite the first layer;

a second heating means thermally coupled to the third layer; and

a continuous channel formed through the first thermally conductive layer, the first insulating layer, the second thermally conductive layer, the second insulating layer and the third thermally conductive layer, said continuous channel having a plurality of cycle segments, each of said cycle segments comprising a first portion disposed within the first thermally conductive layer, a second portion disposed within the third thermally conductive layer and a third portion disposed within the second thermally conductive layer.

35. A device as recited in claim 34 further comprising a fluid movement means coupled to the continuous channel.

36. A device as recited in claim 35 wherein the fluid movement means comprises at least one of a pump or a vacuum pump.

37. A device as recited in claim 34 wherein the first heating means is disposed between the first insulating layer and the first thermally conductive layer.

38. A device as recited in claim 34 wherein the first heating means is part of the first layer.

39. A device as recited in claim 34 wherein the first heating means comprises is coupled to an edge of the first layer.

42. A device as recited in claim 34 further comprising a recirculating device coupled to an inlet port and an outlet port of the continuous channel.

43. A device as recited in claim 42 wherein the recirculating device comprises an upper chamber coupled to the inlet port and a lower chamber coupled to the outlet port, a pump coupled to the inlet port and a plurality of selectively closable valves fluidically coupling the upper chamber and the lower chamber.

44. A device as recited in claim 34 further comprising an LED and a photo detector disposed within the continuous channel.

45. A method of performing a reaction comprising:

introducing a sample into a device having a first layer, a second layer, and a third layer;

performing a first portion of a cycle in the first layer at a first temperature;

moving the sample to the third layer;

thereafter, performing a second portion of the cycle in the third layer at a second temperature;

moving the sample to a second layer;

thereafter, performing a third portion of a cycle in the second layer at a third temperature; and

repeatedly performing the first portion, the second portion and the third portion for a predetermined number of cycles to perform the reaction.

46. A method as recited in claim 45 wherein the second temperature is lower than the first temperature and the third temperature is between the first and second temperature.

47. A method as recited in claim 45 wherein the reaction comprises a polymerase chain reaction.

48. A method as recited in claim 45 wherein the first portion is a denaturing portion.

49. A method as recited in claim 47 wherein the second portion comprises an annealing portion.

50. A method as recited in claim 45 wherein the third portion comprises an extension portion.

51. A method as recited in claim 45 wherein moving the sample to the third layer comprises moving the sample to the third layer through the second layer.

52. A method as recited in claim 45 further comprising providing a first insulator between the first layer and the second layer and a second insulator between the second layer and the third layer.

53. A method as recited in claim 45 wherein moving the sample to the third layer comprises moving the sample to the third layer through the second layer, the first insulator and the second insulator.

54. A method as recited in claim 45 wherein introducing a sample comprises introducing a sample separated by bubbles and further comprising facilitating mixing within the sample using the bubbles.

55. A PCR analyzer comprising:

a plurality of thermal cycling elements, each thermal cycling element receiving the fluidic sample comprising,

a first layer;

a second layer adjacent to the first layer;

a third layer adjacent to the second layer opposite the first layer,

a continuous channel formed within the first layer, the second layer and the third layer, said continuous channel having a plurality of cycle segments, each cycle segment comprising a first portion disposed within the first layer, a second portion disposed within the third layer and a third portion disposed within the second layer,

an incubator having a first platen and a second platen for imparting a first temperature and a second temperature on the thermal cycling element.

56. An analyzer as recited in claim 55 further comprising a sample preparation kit for preparing a fluidic sample.

57. An analyzer as recited in claim 55 further comprising a real time detector coupled to the thermal cycling element for determining an amount of reaction within the thermal cycling element.

58. A method of performing a PCR comprising:

providing a thermal cycling element with one or more tests thereon, said thermal cycling element having a first layer, a second layer adjacent to the first layer, a third layer adjacent to the second layer opposite the first layer and a continuous channel formed within the first layer, the second layer and the third layer, the continuous channel having a plurality of cycle segments, each cycle segment comprising a first portion disposed within the first layer, a second portion disposed within the third layer and a third portion disposed within the second layer;

selecting one or more of the tests;

preparing a fluidic sample corresponding to the one or more tests;

placing the fluidic sample within the thermal cycling device;

incubating the thermal cycling device; and

circulating the fluidic sample through the continuous channel so that the fluidic sample enters multiple first portions, multiple second portions and multiple third portions.

59. A method of performing a PCR comprising:

providing a thermal cycling element having a first layer, a second layer adjacent to the first layer, a third layer adjacent to the second layer opposite the first layer and a continuous channel formed within the first layer, the second layer and the third layer, the continuous channel having a plurality of cycle segments, each cycle segment comprising a first portion disposed within the first layer, a second portion disposed within the third layer and a third portion disposed within the second layer;

preparing a fluidic sample having an enzymatic reaction time and an absorption rate;

placing the fluidic sample within the thermal cycling device;

incubating the thermal cycling device; and

circulating the fluidic sample through the continuous channel so that the fluidic sample enters multiple first portions, multiple second portions and multiple third portions, said circulation rate being a function of the absorption rate and the enzymatic reaction time.