Rapid thermal cycling device

• US 5,455,175 A
• Filed: 01/10/1994
• Issued: 10/03/1995
• Est. Priority Date: 06/04/1990
• Status: Expired due to Fees

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah Research Foundation (State of Utah)
• Inventors: Hillyard, David R., Wittwer, Carl T., Ririe, Kirk M.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Ludlow, Jan M.
• Application Number:
  US08/179,969
• Time in Patent Office: 631 Days
• Field of Search: 435/287-291, 435/809, 219/400, 436/91, 436/147, 422/50, 422/104, 422/109, 422/99, 935/85, 935/88, 392/343, 392/410, 392/412
• US Class Current: 435/286.1
• 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: G01N 2030/3084 ovens
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

A thermal cycling device having a sample chamber whose temperature can be rapidly and accurately modulated over a range of temperatures needed to carry out a number of biological procedures, such a the DNA polymerase chain reaction. Biological samples are placed in glass microcapillary tubes and then located inside the sample chamber. A programmable controller regulates the temperature of the sample inside the sample chamber. Once a heating cycle is completed, the controller opens a door to the chamber for venting hot air out and cool ambient air is moved in. Temperature versus time profiles corresponding to optimum denaturation, annealing and elongation temperatures for amplification of DNA are achievable by the present invention.

Claims

1. An apparatus for subjecting a plurality of biological samples to rapid thermal cycling, the apparatus comprising:

means for holding the plurality of biological samples, the means for holding the plurality of biological samples having an interior and an exterior, the means for holding the plurality of biological samples comprising;

a plurality of biological sample containers, each biological sample container having an elongated cylindrical shape and capable of holding a volume of biological material in the range from about 1 μ

l to about 10,000 μ

l and each of the biological sample containers comprising an inner diameter in the range from about 0.02 mm to about 1.0 mm; and

a sample chamber, the sample chamber having a substantially circular shape, the plurality of biological sample containers being positioned in the sample chamber;

an insulative lining positioned on the interior of the means for holding the plurality of biological samples; and

means for heating the interior of the means for holding the plurality of biological samples, the means for heating being positioned within the sample chamber and comprising an incandescent lamp centrally located among the plurality of biological sample containers;

means for moving air into the means for holding the plurality of biological samples;

means for dispersing the air within the means for holding the plurality of biological samples and for creating a circular air flow path within the means for holding the plurality of biological samples, the means for dispersing the air comprising rotating means for dispersing air positioned within the sample chamber;

means for holding at least a portion of each of the plurality of biological sample containers within the sample chamber such that the portion of the biological sample containers within the sample chamber is oriented substantially perpendicular to the circular air flow path and such that the biological sample containers which are farthest from the means for heating are no more than substantially 40% farther from the means for heating than the distance between the means for heating and those biological sample containers which are closest to the means for heating;

means for venting the means for holding the plurality of biological samples to allow the air moved into the means for holding to cool the sample;

control means for operating, and operatively connected to, the means for heating, the means for moving air, the means for dispersing, and the means for venting; and

means for sensing the temperature in the means for holding the plurality of biological samples, the means for sensing being operatively connected to the control means, such that the temperature of the sample can be rapidly and controllably increased and decreased at a rate at least as great as 1.0°

C./second by the control means in response to the temperature sensed by the means for sensing such that the biological sample can be subjected to rapid thermal cycling over a temperature range of at least 30°

C., the means for sensing comprises a thermocouple, the thermocouple having a cylindrical shape and at least partially positioned in the sample chamber and oriented substantially perpendicular to the circular air flow path, the thermal response of the thermocouple being substantially matched to the thermal response of the biological samples held in the biological sample containers.

2. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the means for holding the plurality of biological samples comprises means for holding at least a portion of each of the plurality of biological sample containers within the sample chamber such that the biological sample containers which are farthest from the means for heating are no more than substantially 25% farther from the means for heating than the distance between the means for heating and those biological sample containers which are closest to the means for heating.

3. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the means for sensing is operatively connected to the control means such that the temperature of the sample can be rapidly and controllably increased and decreased at a rate at least as great as 4.0°

C./second by the control means in response to the temperature sensed by the means for sensing over a temperature range of at least 40°

C.

4. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the means for sensing is operatively connected to the control means such that the temperature of the sample can be rapidly and controllably increased and decreased at a rate at least as great as 10.0°

C./second by the control means in response to the temperature sensed by the means for sensing over a temperature range of at least 40°

C.

5. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the means for venting comprises a door positioned at an upper portion of the means for holding the plurality of biological samples.

6. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 further comprising a housing containing the means for holding the plurality of biological samples, the means for heating, and the means for venting.

7. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the sample containers comprise capillary tubes.

8. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the sample chamber comprises an interior wall, the interior wall being at least partially comprised of a material which converts at least some electromagnetic radiation striking thereon into heat in the sample chamber.

9. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 8 wherein the insulative lining comprises an insulative foam material.

10. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 1 wherein the means for moving air into the means for holding the plurality of biological samples comprises a rotating structure within the means for holding the plurality of biological samples.

11. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 10 wherein the means for moving air into the means for holding the plurality of biological samples creates air velocities in the means for holding the plurality of biological samples at least as great as 200 meters per minute.

12. An apparatus for subjecting a biological sample to rapid thermal cycling as defined in claim 10 wherein the means for moving air into the means for holding the plurality of biological samples further comprises a first portal formed in the means for holding the plurality of biological samples and a larger second portal formed in the means for holding the plurality of biological samples.