Apparatus and method for thermally cycling samples of biological material with substantial temperature uniformity

• US 7,005,617 B2
• Filed: 10/23/2003
• Issued: 02/28/2006
• Est. Priority Date: 07/30/1999
• Status: Expired due to Term

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Stratagene California (Agilent Technologies, Inc.)
• Inventors: Brown, Larry Richard
• Primary Examiner(s): Jeffery, John A.
• Application Number:
  US10/691,874
• Publication Number:
  US 20040149725A1
• Time in Patent Office: 859 Days
• Field of Search: 219476-479, 219/521, 219/530, 219/540, 219/385, 219/430, 422/99, 422/104, 435/285.1, 435/288.4, 435/303.1, 935/85, 935/88, 62/33
• US Class Current: 219/476
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• 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 7/54 using spatial temperature gradients

Abstract

An apparatus for thermally cycling samples of a biological material including a thermal block assembly including a plurality of sample holders for receiving samples of biological material; a heat sink thermally coupled to the thermal block assembly, the heat sink transferring heat away from the thermal block assembly to ambient air in contact with the heat sink; a first heat source thermally coupled to the thermal block assembly to provide heat to the thermal block assembly; and a second heat source thermally coupled to the first heat source and configured to provide heat to a portion of the first heat source. The arrangement of the heat sink, first heat source and second heat source can provide substantial temperature uniformity among the plurality of sample holders. The invention also includes a method for thermally cycling samples of biological material.

Claims

1. An apparatus for thermally cycling samples of biological material comprising:

a thermal block assembly including a plurality of sample holders for receiving samples of biological material;

a heat sink located below the thermal block assembly and thermally coupled to the thermal block assembly to transfer heat away from the thermal block assembly;

a first heat source located between the thermal block assembly and the heat sink, the first heat source thermally coupled to the thermal block assembly to heat the thermal block assembly; and

a second heat source located below the first heat source, wherein the second heat source is radially outside the first heat source, the second heat source thermally coupled to the first heat source to heat at least a portion of the first heat source.

2. The apparatus of claim 1 wherein a stacked arrangement of the first heat source, the second heat source and the heat sink provides substantial temperature uniformity among the plurality of sample holders.

3. The apparatus of claim 1 wherein the first heat source includes at least one thermoelectric heater utilizing the Peltier effect for heating the thermal block assembly with substantial temperature uniformity by heating at least a portion adjacent the edges of the thermal block assembly.

4. The apparatus of claim 1 wherein the first heat source is located on an outer surface of the heat sink causing a temperature gradient across the heat sink.

5. The apparatus of claim 1 wherein the second heat source is located adjacent to at least a portion of the heat sink radially outside of a portion on which the first heat source is located.

6. The apparatus of claim 1 wherein a substantial portion of the second heat source is located outside the first heat source.

7. The apparatus of claim 1 wherein the second heat source includes at least one resistive element heater.

8. The apparatus of claim 1 wherein the first heat source has a higher temperature side and a lower temperature side, the higher temperature side having a higher temperature at an outer periphery of the first heat source than at an inner periphery of the first heat source corresponding approximately to a temperature gradient across the heat sink.

9. The apparatus of claim 1 further comprising a first thermal interface element located between the thermal block assembly and the first heat source to transfer heat to the thermal block assembly.

10. The apparatus of claim 1 further comprising a second thermal interface element located between the heat sink and a lower temperature side of the first heat source to transfer heat and extend the cycle life of the first heat source.

11. The apparatus of claim 1 wherein the thermal block assembly further comprises a thermal block plate, the plurality of sample holders engaging the thermal block plate.

12. The apparatus of claim 1 wherein the plurality of sample holders comprise a plurality of sample wells.

13. The apparatus of claim 1 further comprising a spacer bracket wherein the first heat source is positioned in an opening in the spacer bracket.

14. The apparatus of claim 1 further comprising a first insulating cover to thermally insulate the plurality of sample holders of the thermal block assembly.

15. The apparatus of claim 1 further comprising a third heat source including a plate located above the thermal block assembly to heat a plurality of sample tubes respectively located in the plurality of sample holders of the thermal block assembly.

16. The apparatus of claim 1 further comprising a second insulating cover to thermally insulate a plurality of sample tubes respectively located in the plurality of sample holders of the thermal block assembly.

17. The apparatus of claim 16 wherein the insulating second cover comprises a holding assembly for holding the sample tubes in the thermal block assembly by imparting a compressive load to improve the contact surface area between the respective heat sources and the thermal block assembly, and an insulating plate for the thermal block assembly and a first insulating cover.

18. The apparatus of claim 17 wherein the holding assembly of the second insulating cover includes a bracket with a clamping portion located adjacent the second cover for imparting the compressive load on the insulating plate.

19. The apparatus of claim 1 wherein the biological material includes a biological reaction mixture.

20. The apparatus of claim 1 wherein the second heat source extends beyond the first heat source toward an edge of the thermal block assembly.

21. The apparatus of claim 1 wherein the apparatus is capable of thermally cycling the samples of biological material with substantial temperature uniformity.

22. The apparatus of claim 1 wherein the first heat source is radially separated from the second heat source.

23. An apparatus for thermally cycling samples of biological material with substantial temperature uniformity comprising:

a thermal block assembly including a plurality of sample holders for receiving samples of biological material;

a first heat source located below the thermal block assembly and thermally coupled to the thermal block assembly to heat the thermal block assembly;

a second heat source located below the first heat source, wherein the second heat source is radially outside the first heat source toward an edge of the thermal block assembly, the second heat source thermally coupled to the first heat source to heat at least a portion of the first heat source; and

a heat sink located below the second heat source and thermally coupled to the thermal block assembly to transfer heat away from the thermal block assembly.

24. The apparatus of claim 23 wherein a stacked arrangement of the first heat source, the second heat source and the heat sink provides substantial temperature uniformity among the plurality of sample holders.

25. The apparatus of claim 23 wherein the first heat source includes at least one thermoelectric heater utilizing the Peltier effect for heating the thermal block assembly with substantial temperature uniformity by heating at least a portion adjacent the edges of the thermal block assembly.

26. The apparatus of claim 23 wherein the first heat source is located on an outer surface of the heat sink causing a temperature gradient across the heat sink.

27. The apparatus of claim 23 wherein the second heat source is located adjacent to at least a portion of the heat sink radially outside of a portion on which the first heat source is located.

28. The apparatus of claim 23 wherein a substantial portion of the second heat source is located outside the first heat source.

29. The apparatus of claim 23 wherein the second heat source includes at least one resistive element heater.

30. The apparatus of claim 23 wherein the first heat source has a higher temperature side and a lower temperature side, the higher temperature side having a higher temperature at an outer periphery of the first heat source than at an inner periphery of the first heat source corresponding approximately to a temperature gradient across the heat sink.

31. The apparatus of claim 23 further comprising a first thermal interface element located between the thermal block assembly and the first heat source to transfer heat to the thermal block assembly.

32. The apparatus of claim 23 further comprising a second thermal interface element located between the heat sink and a lower temperature side of the first heat source to transfer heat and extend the cycle life of the first heat source.

33. The apparatus of claim 23 wherein the thermal block assembly further comprises a thermal block plate, the plurality of sample holders engaging the thermal block plate.

34. The apparatus of claim 23 wherein the plurality of sample holders comprise a plurality of sample wells.

35. The apparatus of claim 23 further comprising a spacer bracket wherein the first heat source is positioned in an opening in the spacer bracket.

36. The apparatus of claim 23 further comprising a first cover of insulating material to thermally insulate the plurality of sample holders of the thermal block assembly.

37. The apparatus of claim 23 further comprising a third heat source including a plate located above the thermal block assembly to heat a plurality of sample tubes respectively located in the plurality of sample holders of the thermal block assembly.

38. The apparatus of claim 23 further comprising a second cover of insulating material to thermally insulate a plurality of sample tubes respectively located in the plurality of sample holders of the thermal block assembly.

39. The apparatus of claim 38 wherein the second cover comprises a holding assembly for holding the sample tubes in the thermal block assembly by imparting a compressive load to improve the contact surface area between the respective heat sources and the thermal block assembly, and an insulating plate for the thermal block assembly and a first cover.

40. The apparatus of claim 39 wherein the holding assembly of the second cover includes a bracket with a clamping portion located adjacent the second cover for imparting the compressive load on the insulating plate.

41. The apparatus of claim 23 wherein the biological material includes a biological reaction mixture.

42. The apparatus of claim 23 wherein the first heat source is radially separated from the second heat source.

43. An apparatus for thermally cycling samples of biological material comprising:

a thermal block assembly including at least one sample holder for receiving a biological material;

a heat sink located below the thermal block assembly and thermally coupled to the thermal block assembly to transfer heat from the thermal block assembly;

an upper heat source located under the thermal block assembly, the upper heat source providing heat to the thermal block assembly; and

a lower heat source providing heat to at least a portion of the upper heat source, the lower heat source located under the upper heat source and radially outside the upper heat source.

44. The apparatus of claim 43 wherein the lower heat source is radially separated from the upper heat source.

45. The apparatus of claim 43 wherein a stacked arrangement of the upper heat source, the lower heat source and the heat sink provides substantial temperature uniformity to at least one sample holder.

46. The apparatus of claim 43 wherein the upper heat source includes at least one thermoelectric heater for heating the thermal block assembly.

47. The apparatus of claim 43 further comprising a third heat source including a plate located above the thermal block assembly to heat at least one sample holder.