Thermal cycling apparatus

• US 8,003,370 B2
• Filed: 12/15/2008
• Issued: 08/23/2011
• Est. Priority Date: 05/17/2006
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Goodwin, David, Scherer, Axel, Walker, Christopher I., Maltezos, George, Johnston, Matthew
• Primary Examiner(s): WOOLWINE, SAMUEL C
• Application Number:
  US12/335,448
• Publication Number:
  US 20090275113A1
• Time in Patent Office: 981 Days
• Field of Search: None
• US Class Current: 435/283.1
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 with covers or lids
• 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
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters

Abstract

This invention provides an apparatus for providing heating and cooling of samples in sample vessels having uniform temperature profiles. The apparatus can be used for performing PCR, and real time PCR in particular, with control and uniformity. The system employs a heat block containing a liquid composition to efficiently transfer heat to and from reaction vessels.

Claims

1. A thermal cycler apparatus for performing a polymerase chain reaction (PCR) comprising:

a heat block comprising;

i. an upper surface comprising a plurality of wells in the upper surface adapted to mate with a plurality of sample vessels;

ii. a sealed container comprising an oil in thermal contact with the plurality of wells;

iii. a stirring element in contact with the oil for circulating the oil within the container; and

iv. a bottom surface; and

a heating element in contact with the bottom surface of the heat block.

2. The apparatus of claim 1 further comprising a cooling element in thermal contact with the oil.

3. The apparatus of claim 2 wherein the heating element and the cooling element are Peltier devices.

4. The apparatus of claim 2 wherein the heating element is a resistive heater and the cooling element is a Peltier device.

5. The apparatus of claim 2 wherein the heating element and the cooling element are the same element.

6. The apparatus of claim 5 wherein the same element is a Peltier device.

7. The apparatus of claim 1 further comprising optical assembly having a light source and an optical detector, wherein the optical assembly is positioned such that light from the light source is directed into the sample vessels, and light from the sample vessels is detected by the detector.

8. The apparatus of claim 7 wherein said optical assembly comprises a PIN photodiode, a CCD imager, a CMOS imager, a line scanner, a photodiode, a phototransistor, a photomultiplier or an avalanche photodiode.

9. The apparatus of claim 7 wherein the light source comprises one or more LEDs, laser diodes, vertical cavity surface emitting lasers (VCSELs), vertical external cavity surface emitting lasers (VECSELs), or diode pumped solid state (DPSS) lasers.

10. The apparatus of claim 7 wherein the optical detector comprises a plurality of optical detectors, wherein at least one of the plurality of optical detectors corresponds to each of the plurality of sample vessels.

11. The apparatus of claim 7 further comprising a control assembly which controls the heating element, the light source and the detector.

12. The apparatus of claim 11 wherein the control assembly comprises a programmable computer programmed to automatically process samples, run multiple temperature cycles, obtain measurements, digitize measurements into data and convert data into charts or graphs.

13. The apparatus of claim 12 wherein the multiple temperature cycles correspond to multiple cycles of nucleic acid amplification.

14. The apparatus of claim 13 wherein the nucleic acid amplification comprises real time PCR.

15. The apparatus of claim 1 wherein the container in the heat block comprises a closed system.

16. The apparatus of claim 15 wherein the oil is sealed within the container.

17. The apparatus of claim 1 wherein the stirring element comprises a stir bar.

18. The apparatus of claim 17 wherein the stir bar is a horizontal stir bar.

19. The apparatus of claim 17 wherein the stir bar is a fan shaped stir bar.

20. The apparatus of claim 17 wherein the stir bar has multiple projections.

21. The apparatus of claim 17 wherein the stir bar is magnetically responsive.

22. The apparatus of claim 17 wherein the stir bar is mechanically connected to a motor.

23. The apparatus of claim 1 wherein the stirring element comprises more than one stir bar.

24. The apparatus of claim 1 further comprising a plate having 6, 12, 16, 24, 48, 96, or 384 sample vessels.

25. The apparatus of claim 1 wherein the oil comprises silicone oil or carboxy-modified silicone oil, mineral oil, napthenic oil, hydroisomerized oil, or paraffinic oil.

26. The apparatus of claim 1 wherein the oil comprises a hydrocarbon compound.

27. A heat block for uniformly heating and cooling multiple sample vessels within a sample plate comprising:

a. a receptacle barrier having a receptacle barrier top surface wherein the receptacle barrier top surface comprises wells which correspond to the sample vessels within the sample plate such that when the plate mates with the receptacle barrier, the sample vessels are in thermal contact with the wells; and

b. a base plate, wherein the heat block comprises a liquid container having a top and a bottom and containing a liquid oil;

wherein the receptacle barrier comprises the top of the liquid container, and the base plate comprises the bottom of the liquid container, such that when the sample plate is mated with the receptacle barrier, the liquid oil is in thermal contact with the sample vessels; and

wherein the liquid container comprises a stirring element for circulating the liquid oil within the liquid container; and

c. a heating element in thermal contact with the liquid oil in the liquid container, wherein the heating element is positioned below the base plate outside of the liquid container.

28. The heat block of claim 27 further comprising a cooling element in thermal contact with the liquid oil in the liquid container.

29. The heat block of claim 28 wherein the heating element and the cooling element are the same device.

30. The heat block of claim 29 wherein the same device comprise a Peltier device.

31. The heat block of claim 30 wherein the Peltier device is in physical contact with the base plate.

32. The heat block of claim 27 wherein the heating element comprises a resistive heater.

33. The heat block of claim 27 wherein the sample vessels have a maximum volume of 50 μ

L.

34. The heat block of claim 27 wherein the plate comprises 6, 12, 16, 24, 48, 96, or 384 sample vessels.

35. The heat block of claim 27 comprising a closed system.

36. The heat block of claim 35 wherein the liquid oil is sealed in the heat block during the heating and cooling of the sample vessels.

37. The apparatus of claim 27 wherein the stirring element comprises a stir bar.

38. The apparatus of claim 35 wherein the stir bar is a horizontal stir bar.

39. The apparatus of claim 37 wherein the stir bar is a fan shaped stir bar.

40. The apparatus of claim 37 wherein the stir bar has multiple projections.

41. The apparatus of claim 37 wherein the stir bar is magnetically coupled to a motor.

42. The apparatus of claim 37 wherein the stir bar is mechanically connected to a motor.

43. The apparatus of claim 27 wherein the stirring element comprises more than one stir bar.

44. The apparatus of claim 43 wherein the stir bars are magnetically coupled to one or more motors.

45. The apparatus of claim 43 wherein the stir bars are mechanically connected to one or more motors.

46. The apparatus of claim 27 wherein the base plate comprises a metal plate.

47. The apparatus of claim 46 wherein the base plate comprises aluminum or copper.

48. The heat block of claim 27 wherein the oil comprises silicone oil or carboxy-modified silicone oil, mineral oil, napthenic oil, hydroisomerized oil, or paraffinic oil.

49. The heat block of claim 27 wherein the oil comprises a hydrocarbon compound.