Miniaturized flow thermocycler

• US 5,716,842 A
• Filed: 05/23/1996
• Issued: 02/10/1998
• Est. Priority Date: 09/30/1994
• Status: Expired due to Term

• Current Assignee: Biometra biomedizinische Analytik GmbH (Noblex GmbH)
• Original Assignee: Biometra biomedizinische Analytik GmbH (Noblex GmbH)
• Inventors: Poser, Siegfried, Kohler, Johann Michael, Dillner, Ulrich, Baier, Volker, Bodner, Ulrich, Schimkat, Dieter
• Primary Examiner(s): Redding, David A.
• Application Number:
  US08/649,632
• Time in Patent Office: 628 Days
• Field of Search: 435/283.1, 435/289.1, 435/293.1, 435/305.1, 435/305.4, 422/109, 422/198, 422/68.1, 422/82.11, 422/63, 422/102, 422/58
• US Class Current: 435/283.1
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00873 Heat exchange
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/1883 using thermal insulation
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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: B01L 7/525 with physical movement of samples between temperature zones

Abstract

A miniaturized thermocycler is provided for carrying out thermally controlled biochemical or biological melecular processes, in particular polymerase chain reactions. The aim of the invention is to provide a miniaturized thermocycler which enables such reactions to be carried out more effectively, avoids the problem of parasitic heat absorbers and can be manufactured inexpensively in series. This aim is achieved by virtue of the fact that the sample holder (1) is designed as a series of meanders winding in a plane, the sample holder (1) has a groove in a wall and closed over by a cover, the groove passing alternately through comparable heating zones (2) and cooling zones (3), located at intervals along the groove, as it meanders.

Claims

1. A miniaturized flow thermocycler comprising:

a substrate plate having grooves formed in a plane;

a cover plate having a first surface for covering said substrate plate and sealing said grooves;

means for interconnecting said grooves to form a meandering sample holding channel for receiving and passing liquid media in one direction; and

means for applying heating and means for applying cooling to alternating portions of said meandering sample holding channel such that said meandering sample holding channel is alternatingly affected creating a plurality of alternating heating zones and cooling zones for said liquid media which are spaced apart from one another to effect alternating heating and cooling of said liquid media as said liquid media passes continuously through said meandering sample holding channel in said one direction whereby said meandering sample holding channel functions as a continuous reaction chamber having alternating heating and cooling areas.

2. A miniaturized flow thermocycler according to claim 1, wherein:

said substrate plate having said grooves is formed of one of a silicon and a glass, and said grooves are etched in parallel to one another; and

said interconnecting means includes bridging channels formed in said cover to interconnect ends of said grooves to form said meandering sample holding channel.

3. A miniaturized flow thermocycler according to claim 2, wherein:

said cover is formed of a silicon sheet;

said cover has at least one thermally insulating notch formed in a second surface opposite said first surface to form a diaphragm in said cover for sealing portions of said grooves below said diaphragm; and

said at least one thermally insulating notch being interposed between said means for applying cooling and said means for applying heating thereby serving to thermally isolate said means for applying cooling and said means for applying heating.

4. A miniaturized flow thermocycler according to claim 1, wherein at least one of said heating zones and said cooling zones comprise equally long sample flow sections.

5. A miniaturized flow thermocycler according to claim 1, wherein at least one of the heating zones is divided into spaced apart ranges being subject to different temperatures.

6. A miniaturized flow thermocycler according to claim 1, wherein:

said cover is formed of a silicon sheet;

said cover has at least one thermally insulating notch formed in a second surface opposite said first surface to form a diaphragm in said cover for sealing portions of said grooves below said diaphragm; and

said at least one thermally insulating notch being interposed between said means for applying cooling and said means for applying heating thereby serving to thermally isolate said means for applying cooling and said means for applying heating.

7. A miniaturized flow thermocycler apparatus, comprising:

a substrate assembly having a meandering reaction channel in a plane for receiving a liquid media to be passed therethrough in one direction;

said substrate assembly having at least one thermally insulating notch formed in a first surface thereof and substantially parallel to said plane of said meandering reaction channel to form diaphragm portions between a bottom surface of said notch and said meandering reaction channel; and

said thermally insulating notch dividing said first surface of said substrate assembly into at least one heating area and one cooling area for applying and removing heat from said meandering reaction channel, respectively, such that said meandering channel has alternating heating and cooling portions separated from one another by portions of said meandering reaction channel defined by said diaphragm portions to permit alternating heating and cooling of said liquid media as said liquid media passes through said meandering reaction channel in said one direction.

8. The miniaturized flow thermocycler apparatus according to claim 7 wherein said substrate assembly comprises:

a base member having parallel grooves formed therein;

a cover member covering said base member;

said cover member having interconnect grooves formed in a first surface of said cover member, said first surface contacting and sealing said parallel grooves, and said interconnect grooves interconnecting said parallel grooves at ends thereof to form said meandering reaction channel; and

said cover member having said thermally insulating notch formed in a second surface opposite said first surface, said second surface corresponding to said first surface of said substrate assembly, and said thermally insulating notch being formed perpendicular relative to said parallel grooves.

9. The miniaturized flow thermocycler apparatus according to claim 7 wherein said at least one heating area of said first surface has a heating element formed thereon.

10. The miniaturized flow thermocycler apparatus according to claim 7 wherein at least one of said at least one heating area of said first surface is subdivided by a second thermally insulating notch into two first and second heating areas for heating said liquid media to first and second temperatures respectively.

11. A miniaturized flow thermocycler apparatus, comprising:

a substrate assembly having meandering reaction channel in a plane for receiving a liquid media to be passed therethrough in one direction;

said substrate assembly including a cover member having a first surface covering and sealing said meandering reaction channel;

said cover member having thick areas and thin areas defined by at least one recess formed in a second surface of said cover member opposite said first surface, said thin areas thermally insulating said thick areas from one another;

said thick areas and said thin areas being alternatingly disposed along a path of and adjacent said meandering reaction channel; and

alternating ones of said thick areas along said path of said meandering reaction channel being heating areas and cooling areas, respectively, for applying and removing heat from said meandering reaction channel, respectively, such that said meandering reaction channel has alternating heating and cooling portions separated from one another by portions of said meandering reaction channel adjacent said thin areas to permit alternating heating and cooling of said liquid media as said liquid media passes through said meandering reaction channel in said one direction.