Container for carrying out and monitoring biological processes

• US 20050064582A1
• Filed: 08/09/2004
• Published: 03/24/2005
• Est. Priority Date: 06/04/1990
• Status: Active Grant

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah Research Foundation (State of Utah)
• Inventors: Wittwer, Carl T., Ririe, Kirk M.
• Granted Patent: US 7,745,205 B2
• Time in Patent Office: Days
• US Class Current: 435/287.200
• CPC Class Codes: B01D 15/34 Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 3/5082 Test tubes per se
• 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: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2527/107 Temperature of melting, i.e. Tm
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C12Q 2561/113 Real time assay
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/07 Centrifugal type cuvettes G01N21/09 takes precedence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/06 : Preparation
• CPC Class Codes: G01N 30/466 : with separation columns in parallel
• CPC Class Codes: G01N 35/025 : having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: Y10S 435/808 : Optical sensing apparatus
• CPC Class Codes: Y10S 435/809 : Incubators or racks or holders for culture plates or containers

Abstract

A thermal cycling method and device is disclosed. The device comprises 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 as the DNA polymerase chain reaction. Biological samples are placed in containers each comprising a reservoir and a reaction portion, wherein the reaction portion has a small volume. The small volume reaction portion permits the rapid and accurate temperature modulation. With an optically transmissible reaction portion, DNA amplification may be monitored by fluorescence during PCR.

Claims

1-18. -18. (Cancelled)

19. A container for holding a fluidic biological sample, said container comprising:

a capillary tube portion which is closed at one end; and

a funnel-shaped portion joined to the open end of the capillary tube portion wherein said container holds less than 1 ml of the sample and comprises an optically clear material and wherein said container is adapted for use in nucleic acid amplification using a polymerase chain reaction.

20. The container according to claim 19, wherein said funnel-shaped portion is made from plastic.

21. The container according to claim 19, wherein the capillary tube has a thermal conductivity of from 20-35 in accordance with the formula:

22. The container according to claim 19, wherein said capillary tube has a tip with a curvature selected from the group consisting of an outside curvature and an inside curvature and combinations thereof.

23. The container according to claim 19, wherein said capillary tube portion provides total internal reflection along the length of the capillary tube portion.

24. The container according to claim 19, further comprising a structure to seal the capillary tube portion.

25. The container according to claim 24, wherein said structure is a stopper.

26. The container according to claim 19, wherein said capillary tube portion holds a sample of about 0.01 μ

l to about 100 μ

l.

27. The container according to claim 19, wherein the sample loaded in the capillary tube portion forms a 0.2-2.0 cm column of fluid.

28. A system comprising multiple containers according to claim 19, 34, or 39, and means for holding a plurality of sample containers.

29. A system according to claim 28, wherein said means is a rotatable carousel.

30. A system according to claim 28, further comprising means for performing fluorescent detection in real time.

31. A system according to claim 19, 34, or 39, further comprising an epifluorescent excitation and detection pathway.

32. A system according to claim 31, wherein the detection pathway is focused at the tip of the capillary tube portion.

33. A system according to claim 32, wherein the tip of the capillary tube portion is positioned at the focal point of the detection pathway during monitoring.

34. A container for holding a fluidic biological sample, said container comprising:

a capillary tube portion which is closed at one end;

a funnel-shaped portion joined to the open end of the capillary tube portion; and

a stopper to seal the capillary tube portion wherein said container holds less than 1 ml of the sample and comprises an optically clear material.

35. The container according to claim 34, wherein said funnel-shaped portion is made from plastic.

36. The container according to claim 34, wherein the capillary tube has a thermal conductivity of from 20-35 in accordance with the formula:

37. The container according to claim 34, wherein said capillary tube has a tip with a curvature selected from the group consisting of an outside curvature and an inside curvature and combinations thereof.

38. The container according to claim 34, wherein said capillary tube portion provides total internal reflection along the length of the capillary tube portion.

39. A container holding a fluidic biological sample, comprising:

a capillary tube portion which is closed at one end;

a funnel-shaped portion joined to the open end of the capillary tube portion; and

the biological sample wherein the sample comprises nucleic acids and components for amplification of the nucleic acids using a polymerase chain reaction wherein said container holds less than 1 ml of the sample and comprises an optically clear material.