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EP1000661A1 - Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage - Google Patents

Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage Download PDF

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Publication number
EP1000661A1
EP1000661A1 EP98120187A EP98120187A EP1000661A1 EP 1000661 A1 EP1000661 A1 EP 1000661A1 EP 98120187 A EP98120187 A EP 98120187A EP 98120187 A EP98120187 A EP 98120187A EP 1000661 A1 EP1000661 A1 EP 1000661A1
Authority
EP
European Patent Office
Prior art keywords
ultrathin
multiwell plate
plate according
walled
volume
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP98120187A
Other languages
German (de)
English (en)
Inventor
Alexandre Tretiakov
Hans-Peter Saluz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hans-Knoll-Institut fur Naturstoff-Forschung Ev
Leibniz Institut fuer Naturstoff Forschung und Infektionsbiol eVi
Original Assignee
Hans-Knoll-Institut fur Naturstoff-Forschung Ev
Leibniz Institut fuer Naturstoff Forschung und Infektionsbiol eVi
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hans-Knoll-Institut fur Naturstoff-Forschung Ev, Leibniz Institut fuer Naturstoff Forschung und Infektionsbiol eVi filed Critical Hans-Knoll-Institut fur Naturstoff-Forschung Ev
Priority to EP98120187A priority Critical patent/EP1000661A1/fr
Priority to PCT/EP1999/008178 priority patent/WO2000025920A1/fr
Priority to DE69914220T priority patent/DE69914220T2/de
Priority to EP99952630A priority patent/EP1133359B1/fr
Priority to JP2000579350A priority patent/JP4538152B2/ja
Priority to CA002348564A priority patent/CA2348564A1/fr
Priority to AT99952630T priority patent/ATE257743T1/de
Publication of EP1000661A1 publication Critical patent/EP1000661A1/fr
Priority to US10/848,608 priority patent/US20040214315A1/en
Withdrawn legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B01L3/50851—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates specially adapted for heating or cooling samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples

Definitions

  • the invention relates to plastic plates for conventional heat block thermocycling of biological samples, particularly to multiwell plates. More specifically, it relates to ultrathin-walled multiwell plates with an improved heat transfer to small-volume samples. Such plates can be used for rapid temperature cycling of multiple, small-volume samples (i.e. 1-10 ⁇ l) by using heat block thermocyclers with an increased block temperature ramping (i.e. 4° C/second and greater) and standard heated-lid technology for sealing the plates.
  • the tube has a cylindrically shaped upper wall section and a relatively thin (i.e approximately 0.3 mm) conically-shaped lower wall section.
  • the samples as small as 20 ⁇ l are placed in the tubes, the tubes are closed by deformable, gas-tight caps and positioned into similarly shaped conical wells machined in the body of the heat block.
  • the heated cover compresses each cap and forces each tube down firmly into its own well.
  • the heated platen i.e. heated lid
  • the PCR tubes can be put in a two-piece holder (US patent 5,710,381) of an 8x12, 96-well microplate format, which can be used to support the high sample throughput needs with any number between 1 and 96 individual reaction tubes.
  • the thickness of the tube and the minimal sample volume are yet too high to enable rapid heat transfer.
  • the temperature equilibrium between the large sample and the block is reached relatively slow.
  • the sample temperature lags behind the temperature of the heat block of the thermal cycler for 20-40 seconds and the total DNA amplification time of 1.5 hours or more is typical (see Wittwer and Garling, BioTechniques, 10, 76-83, [1991]).
  • the tubes are designed to hold relatively large sample volumes (50-100 ⁇ l) rather than small ones (0.5-10 ⁇ l), the use of small sample volumes in said tubes results in an increased vapor sorption due to the increased free internal surface of the tube.
  • the multiwell thin-walled PCR plates have been commercially introduced as cost effective alternatives to the above mentioned trays of individual tubes.
  • Such modem plates comprise arrays of conically-shaped wells (0.3mm) molded as a single plastic unit. They are manufactured in various formats to meet the needs of particular biomedical tasks (i.e. 24-well to 48-well plates for routine applications and 96-well to 384-well plates for large scale applications, respectively; see also below). Principally, the same heated-lid technology is used for sitting the arrayed conically shaped wells into the similarly tapered and arrayed wells machined in the body of the heating block. The only difference compared to the tube arrays concerns the sealing of the sample containers. The one-piece plates containing the samples are transferred to the sample block of the thermal cycler, upon sealing all the wells of the plates by a single silicon mat or a special sealing film (e.g.
  • the thickness, geometry and material (polypropylene) of the multiwell-PCR plates is the same as used for the above described tubes, both, the efficiency of the heat transfer and the minimal sample size are directly comparable.
  • the modern heat block thermocyclers are capable to change the block temperature at a rate which is theoretically sufficient to perform an exponential amplification in less than 20 minutes (i.e. Primus-96; ramping rate 5° C/second; supplier: MWG-Biotech, Kunststoff, Germany)
  • the heat transfer from the block to the relatively large sample volumes contained in the modern plastic PCR tubes or multiwell PCR plates is inefficient to reach these amplification times. Therefore, such PCR plates cannot be used for parallel rapid thermocycling of multiple samples.
  • High troughput PCR can be achieved by either increasing the number of samples per run, e. g. 384-well plates and/or by reducing the amplification time.
  • the latter has the advantage of a reduced turnaround, a very important aspect in, for example, rapid and cost effective PCR screening of pooled samples when serial rounds of PCR reactions and product analysis have to be performed.
  • the present invention concerns plastic multiwell plates for performing conventional heat block thermocycling of multiple samples. More specifically, it concerns ultrathin-walled multiwell plates for heat block temperature cycling of samples with a wall thickness of not more than 50 microns. Ultrathin-walled multiwell plates are suited for rapid, oil-free, heat block temperature cycling of small-volume samples (i.e. approximately 0.5-10 ⁇ l) whereas the lower limit is given by the reliability of the conventional pipetting systems.
  • Figure 1 illustrates the multiwell plate according to the invention
  • Figure 2 the positioning of the plate in the block of the thermal cycler
  • Figure 3 illustrates the photograph of electrophoretically separated 454-bp fragments of human papillomavirus DNA amplified by means of rapid PCR.
  • thermoforming thin thermoplastic films are, for example, polyolefin films, such as metallocene-catalyzed polyolefin films, copolymer films and cast polypropylene films, such films having a thickness of not more than 50 microns.
  • the multiwell plate is vacuumformed out of a 30-50 micron cast, unoriented polypropylene film.
  • the film is formed into a female" mold comprising a plurality of spaced-apart, conically shaped wells which are machined in the body of a rectangular- or square-array shaped mold.
  • the advantage of vacuumforming into a female" mold is that the thickness of the walls of the formed wells is gradually reduced to 15-20 ⁇ m at the bottoms of the wells.
  • the plastic material polypropylene is compatible with the standard PCR procedures and therefore widely used for injection molding of PCR tubes and/or multiwell plates. In addition, it has a reduced water vapor sorption when compared to other plastics (e.g. polycarbonate).
  • the volume of the wells is not more than 40 ⁇ l, preferably 16 ⁇ l, the height of the wells is not more than 3.5 mm, and the inter-well spacing is not more than 4.5 mm.
  • the number of wells is in the range of 24-96.
  • the handling of the plate (1) containing the multiple wells (2) is facilitated, by a rigid 0.5-1 mm thick plastic frame (3) which is heat bonded to the plate.
  • the thickness of the well walls of the film-formed plate is reduced 10-20 fold when compared to the conventionally injection-molded PCR plates.
  • the heat transfer through the walls of the film-formed plates is 10-20 fold faster when compared to conventional PCR plates.
  • the geometry of the wells enables the positioning of the entire multiwell plate (1) into the heat block (4), i.e. the parts of the multiwell plate project above the top surface of the block.
  • the pressure caused by the heated lid to the conventionally film- or silicon mat-sealed (13: seal) multiwell plate is actually directed to those parts of the multiwell plate which are supported by the top surface of the heat block (4) and not to the thin walls of the plate as it is the case for the PCR tubes or conventional PCR plates.
  • This advantage makes it possibe to increase the sealing pressure of the heated lid several fold when compared to the conventionally used pressure of 30-50 g per well without cracking the conically shaped walls.
  • the tight thermal contact between the extremely thin walls of the wells and the body of the block (4) is achieved automatically by increased air pressure arrising in the sealed wells at elevated temperatures.
  • samples of a volume of as few as 0.5 ⁇ l can be easily amplified without reducing the PCR efficiency.
  • the following example serves to illustrate the invention but should not be construed as a limitation thereof.
  • reaction mixture prepared according to Ting and Manos (PCR protocols, chapter 42 (1990) Eds.: Innes, Gelfand, Sninsky and White, ISBN 0-12-372180-6) and containing 10 4 input DNA copies of human papilloma virus (HPV-18), integrated into the genome of human HeLa cells, was pipetted (3- ⁇ l volume) into the wells of a 36-well ultrathin-walled plate vacuumformed out of a 47-micron thick cast polypropylene film. The samples were sealed by means of a commercial sealing film, and temperature cycled using a conventional Peltier-driven heat-block thermal cycler (ramping rate 4.5° C/second).
  • Incubation times were as follows: Denaturing: 3 seconds at 95°C, annealing time 3 seconds at 55°C, extension time 16 seconds at 72°C, number of cycles: 30; total amplification time 20 minutes.
  • a photograph in Figure 3 demonstrates some results of the amplification of 454-bp long viral DNA fragments (Line 1-5: viral DNA and line 6: molecular weight marker [Lambda-phage DNA, pstI-restriction digest]).
  • Line 1-5 viral DNA
  • line 6 molecular weight marker [Lambda-phage DNA, pstI-restriction digest]

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Laminated Bodies (AREA)
  • Resistance Heating (AREA)
EP98120187A 1998-10-29 1998-10-29 Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage Withdrawn EP1000661A1 (fr)

Priority Applications (8)

Application Number Priority Date Filing Date Title
EP98120187A EP1000661A1 (fr) 1998-10-29 1998-10-29 Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage
PCT/EP1999/008178 WO2000025920A1 (fr) 1998-10-29 1999-10-28 Plaque a plusieurs puits a parois ultrafines pour thermocyclage par bloc thermique
DE69914220T DE69914220T2 (de) 1998-10-29 1999-10-28 Ultradünnwandige mehrfachlochplatte für heizblock-thermozyklen
EP99952630A EP1133359B1 (fr) 1998-10-29 1999-10-28 Plaque a plusieurs puits a parois ultrafines pour thermocyclage par bloc thermique
JP2000579350A JP4538152B2 (ja) 1998-10-29 1999-10-28 熱ブロック熱サイクル用超薄マルチウェルプレート
CA002348564A CA2348564A1 (fr) 1998-10-29 1999-10-28 Plaque a plusieurs puits a parois ultrafines pour thermocyclage par bloc thermique
AT99952630T ATE257743T1 (de) 1998-10-29 1999-10-28 Ultradünnwandige mehrfachlochplatte für heizblock-thermozyklen
US10/848,608 US20040214315A1 (en) 1998-10-29 2004-05-17 Ultrathin-walled multi-well plate for heat block thermocycling

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
EP98120187A EP1000661A1 (fr) 1998-10-29 1998-10-29 Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage

Publications (1)

Publication Number Publication Date
EP1000661A1 true EP1000661A1 (fr) 2000-05-17

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Family Applications (2)

Application Number Title Priority Date Filing Date
EP98120187A Withdrawn EP1000661A1 (fr) 1998-10-29 1998-10-29 Plaque multi-puits ultramince pour le thermocyclage en utilisant un bloc de chauffage
EP99952630A Expired - Lifetime EP1133359B1 (fr) 1998-10-29 1999-10-28 Plaque a plusieurs puits a parois ultrafines pour thermocyclage par bloc thermique

Family Applications After (1)

Application Number Title Priority Date Filing Date
EP99952630A Expired - Lifetime EP1133359B1 (fr) 1998-10-29 1999-10-28 Plaque a plusieurs puits a parois ultrafines pour thermocyclage par bloc thermique

Country Status (6)

Country Link
EP (2) EP1000661A1 (fr)
JP (1) JP4538152B2 (fr)
AT (1) ATE257743T1 (fr)
CA (1) CA2348564A1 (fr)
DE (1) DE69914220T2 (fr)
WO (1) WO2000025920A1 (fr)

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002026384A2 (fr) * 2000-09-29 2002-04-04 Promega Corporation Plaque d'essai a plusieurs cupules et porte-plaque, et procede de montage de la plaque
US6748332B2 (en) 1998-06-24 2004-06-08 Chen & Chen, Llc Fluid sample testing system
US6780617B2 (en) 2000-12-29 2004-08-24 Chen & Chen, Llc Sample processing device and method
WO2004085134A1 (fr) * 2003-03-24 2004-10-07 Agency For Science, Technology And Research Puce en plastique multipuits peu profonde pour multiplexage thermique
WO2008093109A1 (fr) * 2007-02-02 2008-08-07 Advanced Biotechnologies Limited Plaque multipuits améliorée
WO2010089470A1 (fr) * 2009-02-06 2010-08-12 Bio-Rad Pasteur Appareil de validation thermique, ensemble d'un dispositif de traitement thermique d'échantillons biologiques et d'un tel appareil, et procédé de fabrication d'un tel appareil.
US7799521B2 (en) 1998-06-24 2010-09-21 Chen & Chen, Llc Thermal cycling
EP2255010A1 (fr) * 2008-02-20 2010-12-01 Streck Inc. Thermocycleur et récipient à échantillons pour l'amplification rapide de l'adn
EP2404672A1 (fr) * 2010-07-06 2012-01-11 Universiteit Twente Système multi-puits à haut rendement pour la culture de constructions tissulaires 3D in vitro ou in vivo, procédé de production dudit système multi-puits et procédés de préparation des constructions tissulaires 3D à partir de cellules à l'aide dudit système multi-puits
US8802000B2 (en) 2008-08-01 2014-08-12 Bio-Rad Laboratories, Inc. Microplates with ultra-thin walls by two-stage forming
US8936933B2 (en) 2003-02-05 2015-01-20 IQumm, Inc. Sample processing methods
US9005551B2 (en) 1998-06-24 2015-04-14 Roche Molecular Systems, Inc. Sample vessels
US9737891B2 (en) 2011-06-01 2017-08-22 Streck, Inc. Rapid thermocycler system for rapid amplification of nucleic acids and related methods
US10006861B2 (en) 2013-06-28 2018-06-26 Streck, Inc. Devices for real-time polymerase chain reaction

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1045038A1 (fr) * 1999-04-08 2000-10-18 Hans-Knöll-Institut Für Naturstoff-Forschung E.V. Thermocycleur à bloc de régulation rapide
US7347977B2 (en) 2000-06-08 2008-03-25 Eppendorf Ag Microtitration plate
DE102007062441A1 (de) 2007-12-20 2009-06-25 Aj Innuscreen Gmbh Mobiles Schnelltestsystem für die Nukleinsäureanalytik
KR102206856B1 (ko) * 2017-12-11 2021-01-25 (주)바이오니아 중합효소 연쇄반응 시스템
DE102019106699B4 (de) 2019-03-15 2024-01-25 Analytik Jena Gmbh+Co. Kg Vorrichtung und Verfahren zur thermischen Behandlung von Proben

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Cited By (36)

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US7799521B2 (en) 1998-06-24 2010-09-21 Chen & Chen, Llc Thermal cycling
US10022722B2 (en) 1998-06-24 2018-07-17 Roche Molecular Systems, Inc. Sample vessels
US7833489B2 (en) 1998-06-24 2010-11-16 Chen & Chen, Llc Fluid sample testing system
US6748332B2 (en) 1998-06-24 2004-06-08 Chen & Chen, Llc Fluid sample testing system
US9005551B2 (en) 1998-06-24 2015-04-14 Roche Molecular Systems, Inc. Sample vessels
US7337072B2 (en) 1998-06-24 2008-02-26 Chen & Chen, Llc Fluid sample testing system
WO2002026384A3 (fr) * 2000-09-29 2002-08-15 Promega Corp Plaque d'essai a plusieurs cupules et porte-plaque, et procede de montage de la plaque
WO2002026384A2 (fr) * 2000-09-29 2002-04-04 Promega Corporation Plaque d'essai a plusieurs cupules et porte-plaque, et procede de montage de la plaque
US6660232B1 (en) 2000-09-29 2003-12-09 Promega Corporation Multi-well assay plate and plate holder and method of assembling the same
US6964862B2 (en) 2000-12-29 2005-11-15 Chen & Chen, Llc Sample processing device and method
US9662652B2 (en) 2000-12-29 2017-05-30 Chen & Chen, Llc Sample processing device for pretreatment and thermal cycling
US7935504B2 (en) 2000-12-29 2011-05-03 Chen & Chen, Llc Thermal cycling methods
US8148116B2 (en) 2000-12-29 2012-04-03 Chen & Chen, Llc Sample processing device for pretreatment and thermal cycling
US6780617B2 (en) 2000-12-29 2004-08-24 Chen & Chen, Llc Sample processing device and method
US10443050B2 (en) 2003-02-05 2019-10-15 Roche Molecular Systems, Inc. Sample processing methods
US8936933B2 (en) 2003-02-05 2015-01-20 IQumm, Inc. Sample processing methods
US9708599B2 (en) 2003-02-05 2017-07-18 Roche Molecular Systems, Inc. Sample processing methods
US8080411B2 (en) 2003-03-24 2011-12-20 Agency For Science, Technology And Research Shallow multi-well plastic chip for thermal multiplexing
WO2004085134A1 (fr) * 2003-03-24 2004-10-07 Agency For Science, Technology And Research Puce en plastique multipuits peu profonde pour multiplexage thermique
US7442542B2 (en) 2003-03-24 2008-10-28 Agency For Science, Technology And Research Shallow multi-well plastic chip for thermal multiplexing
WO2008093109A1 (fr) * 2007-02-02 2008-08-07 Advanced Biotechnologies Limited Plaque multipuits améliorée
US9034635B2 (en) 2008-02-20 2015-05-19 Streck, Inc. Thermocycler and sample vessel for rapid amplification of DNA
US20110039305A1 (en) * 2008-02-20 2011-02-17 Streck, Inc. Thermocycler and sample vessel for rapid amplification of dna
EP2255010A4 (fr) * 2008-02-20 2011-09-21 Streck Inc Thermocycleur et récipient à échantillons pour l'amplification rapide de l'adn
EP2255010A1 (fr) * 2008-02-20 2010-12-01 Streck Inc. Thermocycleur et récipient à échantillons pour l'amplification rapide de l'adn
US8802000B2 (en) 2008-08-01 2014-08-12 Bio-Rad Laboratories, Inc. Microplates with ultra-thin walls by two-stage forming
FR2941876A1 (fr) * 2009-02-06 2010-08-13 Bio Rad Pasteur Appareil de validation thermique, ensemble d'un dispositif de traitement d'echantillons biologiques et d'un tel appareil, et procede de fabrication d'un tel appareil.
US9221054B2 (en) 2009-02-06 2015-12-29 Bio-Rad Innovations Thermal validation apparatus, assembly including a device for the thermal processing of biological samples and such an apparatus, and method for manufacturing such an apparatus
WO2010089470A1 (fr) * 2009-02-06 2010-08-12 Bio-Rad Pasteur Appareil de validation thermique, ensemble d'un dispositif de traitement thermique d'échantillons biologiques et d'un tel appareil, et procédé de fabrication d'un tel appareil.
AU2009339202B2 (en) * 2009-02-06 2015-04-02 Bio-Rad Europe Gmbh Thermal validation apparatus, assembly including a device for the thermal processing of biological samples and such an apparatus, and method for manufacturing such an apparatus
WO2012005580A1 (fr) * 2010-07-06 2012-01-12 Universiteit Twente Système à multi-puits à haut rendement pour la mise en culture de constructions tissulaires tridimensionnelles in vitro ou in vivo, procédé de production dudit système à multi-puits et procédé de préparation de constructions tissulaires tridimensionnelles à partir de cellules à l'aide dudit système à multi-puits
EP2404672A1 (fr) * 2010-07-06 2012-01-11 Universiteit Twente Système multi-puits à haut rendement pour la culture de constructions tissulaires 3D in vitro ou in vivo, procédé de production dudit système multi-puits et procédés de préparation des constructions tissulaires 3D à partir de cellules à l'aide dudit système multi-puits
US9737891B2 (en) 2011-06-01 2017-08-22 Streck, Inc. Rapid thermocycler system for rapid amplification of nucleic acids and related methods
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US11385178B2 (en) 2013-06-28 2022-07-12 Streck, Inc. Devices for real-time polymerase chain reaction
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ATE257743T1 (de) 2004-01-15
DE69914220T2 (de) 2004-11-11
DE69914220D1 (de) 2004-02-19
EP1133359A1 (fr) 2001-09-19
JP2002528108A (ja) 2002-09-03
WO2000025920A1 (fr) 2000-05-11
JP4538152B2 (ja) 2010-09-08
CA2348564A1 (fr) 2000-05-11
EP1133359B1 (fr) 2004-01-14

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