Method and apparatus for effecting rapid thermal cycling of samples in microtiter plate size

• US 6,482,615 B2
• Filed: 05/02/2001
• Issued: 11/19/2002
• Est. Priority Date: 03/02/2001
• Status: Expired due to Fees

• Current Assignee: Integrated Genetic Devices Limited
• Original Assignee: Integrated Genetic Devices Limited
• Inventors: Tal, Michael, Liran, Yoram, Livneh, Joshua
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/846,182
• Publication Number:
  US 20020123131A1
• Time in Patent Office: 566 Days
• Field of Search: 435/91.2, 435/285.1, 435/286.6, 435/287.2, 435/303.1, 422/110, 422/131, 935/77, 935/78, 935/85, 935/88, 236/3, 236/91.A, 219/635, 219/663, 219/400
• US Class Current: 435/91.2
• 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

A method and apparatus for effecting rapid thermal cycling of samples, by producing a high-velocity air flow through a closed loop flow path, and energizing an electrical heater within the closed loop flow path to heat the air flowing therethrough to a desired temperature. A sample holder is introduced into the closed loop flow path for exposing the sample holder to the high-velocity heated air flowing therethrough for rapidly heating the sample. The sample is rapidly cooled to a desired temperature by de-energizing the electrical heater, and opening an air outlet from the closed loop flow path, while continuing to produce the high-velocity air flow therethrough.

Claims

1. A method of effecting rapid thermal cycling of samples, comprising:

producing a high-velocity laminar air flow through a channel defining a closed loop flow path;

energizing an electrical heater within said closed loop flow path to heat the air flowing therethrough to a desired temperature;

introducing a sample holder containing at least one sample into a section of said closed loop flow path for exposing said sample holder to the high-velocity heated air flowing therethrough for rapidly heating the sample;

and rapidly cooling the sample to a desired temperature by de-energizing said electrical heater and opening an air outlet from said closed loop flow path, while continuing to produce said high-velocity airflow through the channel.

2. The method according to claim 1, wherein said section of the closed loop flow path into which the sample holder is introduced is of rectangular cross-section.

3. The method according to claim 1, wherein said channel defining said closed loop flow path is a closed loop channel having a selectively-openable inlet and a selectively-openable outlet.

4. The method according to claim 1, wherein said sample holder is introduced into a sample-holder compartment including a cover containing another electrical heater which may be energized during the process to reduce excessive vaporization of the sample.

5. The method according to claim 1, wherein the air flow velocity is controlled to produce a temperature gradient across the sample holder.

6. The method according to claim 1, wherein said section of the closed loop flow path into which said sample holder is introduced, has a transverse dimension slightly larger than that of a said sample holder to define an air flow space between said section of the closed loop flow path and said sample holder on each of the opposite sides of the sample holder.

7. The method according to claim 1, wherein the air velocity through said section of the closed loop flow path is 10-30 m/sec.

8. The method according to claim 1, wherein said electrical heater is of annular configuration circumscribing a portion of said closed loop flow path.

9. The method according to claim 1, wherein said sample holder includes a plurality of metal tubes arranged in a rectangular matrix array for receiving the samples.

10. The method according to claim 1, wherein the section of said closed loop flowpath into which the sample holder is introduced is of decreasing cross-sectional area from the upstream side of said sample holder to the downstream side of the sample holder, to produce an increase in the velocity of the airflow at said downstream side as compared to that at said upstream side.

11. The method according to claim 10, wherein said decreasing cross-sectional area is effected by the provision of an inclined baffle in said section of the closed loop flow path into which the sample holder is introduced, said inclined baffle underlying said sample holder.

12. The method according to claim 10, wherein said decreasing cross-sectional area is effected by mounting the sample holder in an inclined position in said section of the closed loop flowpath.

13. Apparatus for effecting rapid thermal cycling of samples, comprising:

a housing including a channel defining a closed loop flow path for air;

an impeller within said housing for producing a high velocity laminar air flow through said closed loop flow path;

an electrical heater within said housing for heating the air flowing through said closed loop flow path to a desired temperature;

an access opening in a section of said channel for introducing a sample holder containing at least one sample into a sample compartment in said closed loop flow path for exposure to the high-velocity heated air flowing therethrough; and

a control means for selectively energizing the electrical heater to rapidly heat said sample by the heated air flowing through the closed loop flowpath, and for selectively de-energizing the electrical heater and opening the closed loop flow path with respect to the atmosphere to rapidly cool said sample.

14. The apparatus according to claim 13, wherein said section of the channel into which the sample holder is introduced is of rectangular cross section.

15. The apparatus according to claim 13, wherein said channel comprises a first section including first and second legs parallel to each other and joined by a first U-shaped juncture, and a second section including third and fourth legs parallel to each other and joined by a second U-shaped juncture;

said first and second legs of the first section being perpendicularly joined to said third and fourth legs of the second section to define a closed loop flow path constituted of two U-shaped loops perpendicularly joined to each other.

16. The apparatus according to claim 13, wherein said sample compartment has a bottom mat and includes a cover for pressing the sample-holder against the mat, the cover containing another electrical heater which may be energized during the process to reduce excessive vaporization of the samples.

17. The apparatus according to claim 13, wherein the section of the channel into which sample holder is introduced has a transverse dimensions slightly larger than that of said sample holder to define an air flowspace between said section of the channel and said sample holder on each of the opposite sides of the sample holder.

18. The apparatus according to claim 13, wherein said channel defining said closed loop flow path is a closed loop channel including an inlet opening leading from the atmosphere into said loop channel, an outlet opening leading from the closed loop channel to the atmosphere, and an actuator for selectively opening and closing at least said outlet opening.

19. The apparatus according to claim 18, wherein there is also an actuator for selectively opening and closing said inlet opening.

20. The apparatus according to claim 13, wherein said electrical heater is of annular configuration circumscribing a portion of said closed loop flow path.

21. The apparatus according to claim 13, wherein said electrical heater is positioned inside the channel upstream from the air impeller.

22. The apparatus according to claim 13, wherein said sample holder includes a plurality of metal tubes arranged in a rectangular matrix array for receiving the samples.

23. Apparatus for effecting rapid thermal cycling of samples, comprising:

a housing including a channel defining a closed loop flow path for air;

an impeller within said housing for producing a high velocity air flow through said closed loop flow path;

an electrical heater within said housing for heating the air flowing through said closed loop flow path to a desired temperature;

an access opening in a section of said channel for introducing a sample holder containing at least one sample into a sample compartment in said closed loop flow path for exposure to the high-velocity heated air flowing therethrough;

and control means for selectively energizing the electrical heater to rapidly heat said sample by the heated air flowing through the closed loop flowpath, and for selectively de-energizing the electrical heater and opening the closed loop flow path with respect to the atmosphere to rapidly cool said sample;

the section of said closed loop flow path into which the sample holder is introduced being of decreasing cross-sectional area from the upstream side of said sample holder to the downstream side of the sample holder, to produce an increase in the velocity of the airflow at said downstream side as compared to that at said upstream side.

24. The apparatus according to claim 23, wherein said decreasing cross-sectional area is effected by including an inclined baffle in said section of the closed loop flow path into which the sample holder is introduced, said baffle underlying said sample holder.

25. The apparatus according to claim 23, wherein said decreasing cross-sectional area is effected by mounted the sample holder in an inclined position in said section of the closed loop flowpath.

26. The apparatus according to claim 23, wherein said impeller produces a high velocity laminar air flow through said closed loop flow path.