Temperature cycling cuvette

• US 4,902,624 A
• Filed: 11/21/1988
• Issued: 02/20/1990
• Est. Priority Date: 11/23/1987
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Helfer, Jeffrey L., Columbus, Richard L., Porte, Johannes J., Wellman, Jeffrey A.
• Primary Examiner(s): Kamen, Noah P.
• Application Number:
  US07/273,781
• Time in Patent Office: 456 Days
• Field of Search: 435/287, 435/292, 435/296, 435/316, 356/246, 220/86 R, 215/1 R, 215/1 C, 422/58, 422/61, 422/102, 422/103, 422/104, 137/588
• US Class Current: 435/285.1
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• 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 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2537/10 the purpose or use of
• CPC Class Codes: C12Q 2565/519 characterised by the capture moiety being a single stranded oligonucleotide

Abstract

There is disclosed a thermal cycling cuvette constructed with an improved sample fluid thermal time constant. The fluid surface-to-volume ratio, and the cuvette thermal path length and thermal resistance of at least one of the walls that provides a major surface of contact, are selected to provide such a thermal time constant that is no greater than about 10 seconds for a cuvette volume no greater than about 200 μl. Also included in the cuvette are means permitting the introduction and removal of liquid into and from the cuvette. A preferred construction permits ready and substantially complete removal of the liquid.

Claims

1. In a thermal cycling cuvette for controlled reaction of components of a liquid involving cycling through a temperature range of at least about 35°

C., said cuvette having at least one liquid-confining chamber defined by two spaced-apart opposing walls each providing a major surface of contact, side walls connecting said two opposing walls, and means permitting the introduction of liquid into, and the removal of such liquid from, said chamber;

the improvement wherein said opposing walls and said side walls are dimensioned to provide (a) a spacing between said opposing walls of no less than 0.5 mm and (b) a predetermined surface-to-volume ratio of said chamber, at least one of said opposing walls being provided with a predetermined thermal path length and thermal resistance such that said ratio, path length and thermal resistance are effective to provide, for pure water contained within said chamber in contact with said opposing walls, a thermal time constant for such water that is no greater than about 10 seconds, for a liquid volume of no more than about 200 μ

l.

5. A cuvette as defined in claim 1 or 3, wherein the structural component of at least one of said opposing walls comprises aluminum no thicker than about 0.15 mm.

6. A cuvette as defined in claim 1 or 3, wherein at least one entire surface of one of said opposing walls of said chamber has a thermal resistance of no more than about 0.01°

C./watt.

7. A cuvette as defined in claim 1 or 3, and further including a reagent coated over at least one of said opposing walls.

8. A cuvette as defined in claim 7, wherein said one opposing wall is opposite to said opposing wall having said thermal path length.

9. A cuvette as defined in claim 1 or 3, wherein said permitting means includes means defining a liquid access aperture.

10. A cuvette as defined in claim 9, and further including an opening constructed to be larger than said liquid access aperture, to receive closure means for sealing said larger opening, and wherein said aperture is located within said larger opening.

11. A cuvette as defined in claim 10, and further including an air vent for said chamber.

12. A cuvette as defined in claim 11, wherein said air vent terminates at a location within said larger opening.

13. A cuvette as defined in claim 9, wherein said liquid access aperture is configured to removably mate with a pipette or pipette tip.

14. A cuvette as defined in claim 1 or 3, wherein said permitting means includes a liquid access aperture, an air vent, and a movable valve for selectively sealing off fluid flow between (a) each of said access aperture and air vent, and (b) said chamber.

15. A cuvette as defined in claim 14, wherein a portion of said valve is rotatably mounted in the cuvette, and further including means for rotating said valve portion about an axis, said access aperture and said air vent being rotationally oriented about said axis to cooperate with rotation of said valve portion to open and close said access aperture and said air vent.

16. A cuvette as defined in claim 14, wherein a portion of said valve is translatably mounted in the cuvette, and further includes biasing means for biasing said valve portion to a position that seals off said chamber from the exterior of the cuvette.

17. A cuvette as defined in claims 1 or 3, and further including a second liquid-confining chamber defined by two spaced-apart opposing walls with a relatively high exposed surface-to-thickness ratio, side walls connecting said second-chamber two opposing walls, means permitting the introduction of liquid into said second chamber from said first chamber, and means permitting the removal of such liquid out of said second chamber;

said chambers further providing a major plane of liquid containment, the major plane of one of said chambers being disposed above the major plane of the other of said chambers.

18. A cuvette as defined in claim 17, and further including a third liquid-confining chamber fluidly connected to said second chamber, and a liquid-permeable membrane separating said third chamber from said second chamber.

19. A cuvette as defined in claim 18, and further including a liquid-absorbing material in said third chamber.

20. A cuvette as defined in claim 18, and further including on said membrane, means for capturing a proteinaceous material capable of bonding to a DNA strand.

21. A cuvette as defined in claim 20, wherein said capturing means includes either avidin or biotin, said proteinaceous material having bonded thereto, the other of the avidin/biotin pair.

22. A cuvette as defined in claim 1 or 3, wherein one of said walls is transparent and is recessed to protect it from scratches.

2. In a thermal cycling cuvette for controlled reaction of components of a liquid involving cycling through a temperature range of at least about 35°

C., said cuvette having at least one liquid-confining chamber defined by two spaced-apart opposing walls, side walls connecting said two opposing walls, and means permitting the introduction and removal of such liquid, into and from said chamber,

the improvement wherein said opposing walls and said side walls are dimensioned to provide said chamber with an effective temperature controlled surface-to-volume ratio that is between about 65 in^-1 and about 130 in^-1 for a fill volume of between 100 and 200 μ

l,

and at least one of said opposing walls having a thermal path length of no more than about 0.3 mm and a thermal resistance of no more than about 0.01°

C./watt,

whereby the cuvette has a maximized rate of heat transfer during heating and cooling cycles.

3. A cuvette as defined in claim 2, wherein said opposing walls and said side walls are spaced so that water can be effectively withdrawn in no longer than 10 seconds with a capillary number of less than 0.05.

4. A cuvette as defined in claim 3, wherein said distance of spacing of said opposing walls is from about 0.5 mm to about 2.5 mm.

23. In a cuvette for controlled reaction of components of a liquid, said cuvette having first and second liquid confining chambers for conducting reactions, each defined by two spaced-apart opposing walls, side walls connecting respective ones of said two opposing walls, and means permitting the introduction and removal of such liquid into and from each of said chambers, each of said chambers providing a major plane of liquid containment;

the improvement wherein said chambers are disposed such that said major plane of one of said chambers lies disposed above the major plane of the other of said chambers;

and further including a third liquid-confining chamber fluidly connected to said second chamber, and a liquid permeable membrane separating said third chamber from said second chamber.

24. A cuvette as defined in claim 23, and further including a liquid-absorbing material in said third chamber.

25. A cuvette as defined in claim 23, and further including on said membrane, means for capturing a proteinaceous material capable of bonding to a DNA strand.

26. A cuvette as defined in claim 25, wherein said capturing means includes either avidin or biotin, said proteinaceous material having bonded thereto, the other of the avidin/biotin pair.