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

• US 6,657,169 B2
• Filed: 07/30/1999
• Issued: 12/02/2003
• Est. Priority Date: 07/30/1999
• Status: Expired due to Term

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Stratagene California
• Inventors: Brown, Larry Richard
• Primary Examiner(s): Jeffery, John A.
• Application Number:
  US09/364,051
• Publication Number:
  US 20020030044A1
• Time in Patent Office: 1,586 Days
• Field of Search: 219/476, 219/477-479, 219/521, 219/530, 219/540, 219/385, 422/99, 422/104, 135/285.1, 135/288.4, 135/303.1, 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 heating samples of biological material, comprising:

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, said 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, said first heat source generally positioned in a stacked relationship between the heat sink and the thermal block assembly; and

a second heat source located below the first heat source with a substantial portion of the second heat source located outside the first heat source and thermally coupled to the first heat source to provide heat to at least a portion of the first heat source,

wherein said thermal block assembly is positioned above the heat sink in said stacked relationship with the heat sink, and further wherein the arrangement of the heat sink, first heat source and second heat source provides substantial temperature uniformity among the plurality of sample holders.

2. The apparatus of claim 1, wherein said second heat source is located on an outer surface of the heat sink in order to cause a temperature gradient across the heat sink.

3. The apparatus of claim 2, wherein said first heat source includes at least one thermoelectric heater utilizing the Peltier effect, said first heat source producing a temperature gradient as a function of the temperature gradient across the heat sink, said temperature gradient of the first heat source causing the thermal block assembly to be urged toward said substantial temperature uniformity by heating at least a portion adjacent the edges of the thermal block assembly.

4. The apparatus of claim 3, wherein said second heat source is located adjacent at least a portion of the heat sink radially outside of the portion on which the first heat source is located.

5. The apparatus of claim 4, wherein said second heat source includes at least one resistive element heater.

6. The apparatus of claim 3, wherein during a first phase of operation, the first heat source and the second heat source work in conjunction to heat the thermal block assembly, and during a second phase of operation, the first heat source operates to direct heat away from the thermal block and into the heat sink.

7. The apparatus of claim 1, said first heat source including a first side and a second side, said first 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.

8. The apparatus of claim 1, wherein said thermal block assembly further comprises a thermal block plate.

9. The apparatus of claim 8, wherein said plurality of sample holders comprise a plurality of sample wells.

10. The apparatus of claim 9, wherein said plurality of sample wells are coupled to a surface of the thermal block plate.

11. The apparatus of claim 10, wherein said plurality of sample wells are attached to the thermal block plate by a method selected from a group consisting of silver brazing, copper casting, and ultrasonic welding.

12. The apparatus of claim 10, wherein said plurality of sample wells and thermal block plate are both composed of a copper alloy.

13. The apparatus of claim 1, wherein said first heat source includes at least one thermoelectric heater utilizing the Peltier effect, said at least one thermoelectric heater including a first side and second side, said first side typically being at a higher temperature than said second side during a heating of the sample, said apparatus further comprising a first thermal interface element located between the thermal block assembly and the first side of the at least one thermoelectric heater to provide heat transfer to the thermal block assembly.

14. The apparatus of claim 13, further comprising a second thermal interface element located between the heat sink and a lower temperature side of the at least one thermoelectric heater to provide heat transfer between the heat sink and the lower temperature side of the at least one thermoelectric heater and to improve the cycle life of the thermoelectric heater.

15. The apparatus of claim 1, wherein said thermal block assembly further comprises a thermal block plate, said plurality of sample holders being located on a surface of the thermal block plate,

said apparatus further comprising:

a first cover of insulating material on the thermal block assembly; and

a plurality of longitudinal fasteners for imparting a compressive force across said first cover, said thermal block plate, said first heat source, and said heat sink.

16. The apparatus of claim 15, wherein each said longitudinal fastener includes a screw and an insulating screw cap on one end of the longitudinal fastener, said insulating screw cap being located on a bottom surface of the heat sink.

17. The apparatus of claim 16, wherein each longitudinal fastener further includes an insulating washer located between the end of each longitudinal fastener and the bottom surface of the heat sink.

18. The apparatus of claim 1, further comprising a spacer bracket, each said first heat source being positioned in an opening in the spacer bracket.

19. The apparatus of claim 18, wherein said spacer bracket is sized so that the thermal block assembly is placed in compression.

20. The apparatus of claim 1, further comprising:

a first cover of insulating material, said first cover thermally insulating the sample holder of the thermal block assembly; and

a third heat source, said third heat source including a plate located above the thermal block assembly to provide heat to a plurality of sample tubes respectively located in the sample wells of the thermal block assembly, wherein said sample holders comprise sample wells.

21. The apparatus of claim 20, further comprising:

a second cover of insulating material, said second cover thermally insulating the plurality of sample tubes respectively located in the sample wells of the thermal block assembly, said second cover comprising an assembly for holding the sample tubes in the thermal block by imparting a compressive load to improve the contact surface area between the respective heat sources and thermal block assembly, and a plate of insulating material for the thermal block assembly and first cover.

22. The apparatus of claim 21, wherein said holding assembly of the second cover includes a bracket with a clamping portion located adjacent the second cover for imparting said compressive load on the insulating plate.

23. The apparatus of claim 1, wherein said biological material includes a biological reaction mixture.

24. The apparatus of claim 23, wherein said apparatus is capable of thermally cycling said samples of biological reaction mixture.

25. The apparatus of claim 1, wherein the first heat source comprises at least one Peltier heater and the second heat source comprises at least one resistive heater.

26. The apparatus of claim 1 wherein the second heat source is located outside the first heat source.

27. An apparatus for heating samples of a biological material, comprising:

a thermal block assembly including a plurality of sample wells for receiving samples of biological material, each sample well having a length defined by the distance between a bottom surface of the sample well and a top edge surface of the sample well;

a first heat source thermally coupled to the thermal block assembly, said first heat source providing heat to the thermal block assembly;

a second heat source located below the first heat source with a substantial portion of the second heat source located outside the first heat source to provide heat to a portion of the first heat source; and

a first cover of insulating material, said first cover thermally insulating the sample wells of the thermal block assembly, said first cover comprising a plate with a plurality of cylindrical sample well openings, each cylindrical sample well opening corresponding to a respective sample well,

wherein the first cover includes a top portion configured to substantially cover the top edge surface of each of the sample wells, the first cover further including a peripheral portion configured to extend around a periphery of the plurality of sample wells and surround the plurality of sample wells along a substantial portion of the length of the sample wells.

28. The apparatus of claim 27, wherein each sample well extends for a substantial length of its respective cylindrical sample well opening in the first cover.

29. The apparatus of claim 27, wherein said thermal block assembly further comprises a thermal block plate, said plurality of sample wells being coupled to a surface of the thermal block plate.

30. The apparatus of claim 29, wherein the thermal block assembly further includes at least one sensor cup coupled to the surface of the thermal block plate, and wherein said first cover further includes at least one sensor cup opening corresponding to a respective sensor cup.

31. The apparatus of claim 27, further comprising:

a third heat source, said third heat source including a plate located above the thermal block assembly to provide heat to a plurality of sample tubes respectively located in a plurality of sample wells of the thermal block assembly.

32. The apparatus of claim 31, wherein said first heat source is a thermoelectric heater utilizing the Peltier effect, said second and third heat sources are resistive element heaters, and said samples of biological material is a biological reaction mixture including DNA.

33. The apparatus of claim 27, further comprising a heat sink thermally coupled to the thermal block assembly, wherein said second heat source is located on an outer surface of the heat sink in order to cause a temperature gradient on the heat sink.

34. The apparatus of claim 27, wherein the peripheral portion of the first cover is configured to surround the sample wells along the entire length of the sample wells.

35. The apparatus of claim 27 wherein the second heat source is located outside the first heat source.

36. An apparatus for thermally cycling samples of biological reaction mixture, comprising:

a thermal block assembly including a thermal block plate and a plurality of sample wells for receiving sample tubes containing biological reaction mixture;

a heat sink thermally coupled to the thermal block assembly, said heat sink transferring heat from the thermal block assembly to ambient air in contact with the heat sink, said thermal block assembly positioned above the heat sink in a stacked relationship with the heat sink;

at least one thermoelectric heater thermally coupled to the thermal block assembly, said at least one thermoelectric heater providing heat to the thermal block assembly, said at least one thermoelectric heater generally positioned in a stacked relationship between the heat sink and the thermal block assembly;

a thermal interface material between outer surfaces of the at least one thermoelectric heater and at least one of the thermal block assembly and heat sink, said thermal interface material providing heat transfer between the at least one thermoelectric heater and the at least one of the thermal block assembly and heat sink;

a first cover of insulating material for the sample wells of the thermal block assembly;

a first resistive element heater, said first resistive heater including a flat plate located above the thermal block assembly to provide heat to the thermal block assembly; and

a second resistive element heater located below the at least one thermoelectric heater with a substantial portion of the second resistive element heater located outside the at least one thermoelectric heater for heating a bottom portion of said at least one thermoelectric heater.

37. The apparatus of claim 36, further comprising a second cover of insulating material including a holding assembly for holding the sample tubes in the sample wells by imparting a substantially uniform compressive load, and a plate of insulating material for the thermal block and first cover.

38. The apparatus of claim 36 wherein the second resistive element heater is located outside the at least one thermoelectric heater.