WO2001014063A1 - Dispositif et procede permettant de manipuler et de traiter des echantillons et/ou des melanges reactionnels de petit volume - Google Patents

Dispositif et procede permettant de manipuler et de traiter des echantillons et/ou des melanges reactionnels de petit volume Download PDF

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Publication number
WO2001014063A1
WO2001014063A1 PCT/SE2000/001632 SE0001632W WO0114063A1 WO 2001014063 A1 WO2001014063 A1 WO 2001014063A1 SE 0001632 W SE0001632 W SE 0001632W WO 0114063 A1 WO0114063 A1 WO 0114063A1
Authority
WO
WIPO (PCT)
Prior art keywords
format
microtitre plate
well
adapter
well format
Prior art date
Application number
PCT/SE2000/001632
Other languages
English (en)
Inventor
Mats Malmquist
Original Assignee
Alphahelix Ab
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 Alphahelix Ab filed Critical Alphahelix Ab
Priority to EP00957213A priority Critical patent/EP1214147A1/fr
Priority to AU68866/00A priority patent/AU6886600A/en
Priority to JP2001518192A priority patent/JP2003531358A/ja
Publication of WO2001014063A1 publication Critical patent/WO2001014063A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/251—Colorimeters; Construction thereof
    • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
    • 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
    • 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
    • B01L3/50855—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 using modular assemblies of strips or of individual wells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N21/07—Centrifugal type cuvettes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/021—Adjust spacings in an array of wells, pipettes or holders, format transfer between arrays of different size or geometry
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0893—Geometry, shape and general structure having a very large number of wells, microfabricated wells

Definitions

  • the present invention concerns a device and a method for the simultaneous handling of a multitude of small volume samples, and in particular novel means and a method for compressing or condensing a set of samples such as a set of samples on a microtitre plate from one format to a less space-demanding one.
  • Biochemical reactions are routinely carried out in so-called microtitre plates - sheets of moulded plastic that typically contain eight rows and twelve columns of tiny wells, each of which capable of holding a few millilitres or less of the reaction rnixture. This is known as the 8 by 12 or 96-well format.
  • the 96-well format is widely used, not only in biochemical operations, such as analysis, diagnostics, small scale synthesis and the like, but also in fields like high throughput screening and combinatorial chemistry.
  • the receiving microtitre plate cannot have too high resolution.
  • a very dense format will become physically unmanageable from the user's point of view and the risk of mix-ups and contamination increases.
  • the 96-well format is widely used and accepted by the laboratories, both in research and other applications. Auxiliary equipment has been designed to accommodate this format.
  • the historical and wide-spread use of the 96-well format means that vast amounts of old patient samples, compounds for drug development etc are stored in the 96-well format.
  • the format has to be compressed. Presently, such reformation, compression or condensation is done by pipetting robots.
  • the above problem is solved by the device and method as defined in the attached claims.
  • the inventive device allows easy conversion between different microtitre plate formats, thus adding flexibility and making possible the sequential use of apparatuses designed for different formats.
  • Other applications and benefits of the invention will be described in or become evident from the description and examples below.
  • FIG. 1 shows schematically the compression steps from the 96-well format to the
  • Fig. 2 shows an adapter according to claim X in perspective
  • Fig. 3 shows the above adapter in cross section along the line IJJ;
  • Fig. 4 shows the above adapter in cross section along line TV; and Fig. 5 shows schematically how four adapters, two of each (denoted A and B, respectively) are assembled together.
  • microtitre plates in general, which is hereby defined as comprising any device capable of receiving a multitude of samples in separate and/or discrete locations or so called wells.
  • grid is used to define the spatial distribution of said wells, e.g. their distribution in rows and columns, on the microtitre plate.
  • upper is used to define the surface of the adapter having a grid corresponding to the less dense format. Consequently, the term “lower” is used to define the surface of the adapter having a grid corresponding to the more dense format
  • small volume reactions includes any chemical reaction conducted in volumes below 100 ⁇ l.
  • Typical biochemical reactions are immunological determinations, histological determinations, enzymatic determinations, biochemical diagnosis of diseases, determinations of paternity, various determinations in forensic medicine, various operations in analysis and synthesis applications.
  • Typical chemical reactions intended to fall within this definition are reactions in the screening and synthesis of pharmaceuticals, for example high throughput screening, combinatorial chemistry, analysis, determination of environmental pollutants etc.
  • the adapter according to the invention is a three-dimensional body having openings on one surface, said openings spaced according to the less dense donating format, and a corresponding amounts of openings on its reverse surface, said openings being connected by individual channels.
  • the relative position of each opening on one surface is identical to the position of the corresponding opening on the reverse surface.
  • the grid of the wells on the upper surface corresponds to the grid of microtitre plates of this less dense format, which is subject of compression.
  • the grid of the openings on the lower surface in its turn corresponds to the grid of the more dense, receiving format.
  • An adapter for compressing the 96-well format into the 384-well format typically has 24 wells, arranged in 6 rows and 4 columns, on its upper surface. Correspondingly, there are 24 openings on its lower surface, arranged in 6 rows and 4 columns.
  • the grid on the upper surface is identical to the grid of a 96-well microtitre plate, and the grid on the lower surface is identical to the grid of a 384-well microtitre plate.
  • the adapter is manufactured in two parts being a mirror image of each other. When put into use, two of each part are assembled as shown in Fig. 3. In order to compress a conventional 8 by 12 microtitre plate (the 96-well format) into a more dense format, e.g. a 16 by 24 plate (the 384-well format) four adapters are assembled as illustrated in Fig. 3. The set of four adapters can be arranged on top of a receiving microtitre plate, e.g. one comer or a quarter of a microtitre plate in the 384-well format. The samples and reagents, as desired, are then dispensed in the wells of the adapters, either manually or using equipment developed for the 96-well format.
  • the adapters together with the receiving microtitre plate or quarter thereof, are then placed in a centrifuge and subjected to a centrifugal force sufficient to transfer all the contents of the upper wells, through the channels, into the receiving denser format.
  • the adapters can then be removed and discarded and the samples, being in the 384-well format, processed further or analysed using equipment adapted to this format.
  • Possible further steps for the contents transferred to the denser format include analysis, storage or emptying or a combination thereof. Analysis steps may include the addition of a further reagent and the analysis itself can encompass photometric determinations, such as measuring the fluorescence intensity of the mixture.
  • the channels connecting the upper and lower openings on an adapter are made so narrow, that the sample and/or reaction mixture cannot pass the channels unless the adapter is subjected to a certain centrifugal force.
  • the channels can be closed by an inert substance which is either solid or viscous at normal analysis temperature, but which melts or turns less viscous at an increased temperature within a range, tolerable for the sample and reagents. In this way the user can have control over the reaction, for example by adding a sample plus reagent in the less dense format and, at a desired moment, end the reaction by centrifuging the mixture into the receiving format, possibly containing an agent quenching the reaction.
  • the channels connecting the upper and lower openings on the adapter are provided with constrictions or filter means for separating the sample and reagent mixture from auxiliary components, such as particulate matter or reagent capillaries.
  • Particulate matter e.g. silica particles, glass beads, latex beads etc is frequently used in extraction, purification and separation operations.
  • An adapter provided with constrictions or filter means would therefor function as a convenient means for separating the particulate matter from the reaction mixture, simultaneously as the separate samples are transferred to a denser format.
  • the filter means constitute a convenient way to simultaneously separate the particulate matter from the reaction mixture and emptying the contents into a denser format for further analysis or subsequent reactions.
  • the adapter itself can function as a microtitre plate, being the receptacle of samples and reagents, with the further benefit of allowing the reaction mixture to be transferred into a denser format at any time desired by the user.
  • the adapter or device according to the invention is preferable made of a suitable thermoplastic.
  • suitable materials include, but are not limited to, polypropylene (PP), polystyrene (PS), polyethylene (PE), high density polyethylene (HDPE), polycarbonate (PC), polyacetate (PA), poly-methyleene-methacrylate (PMMA) and poly-vinylidene-fluoride (PVDF).
  • PP polypropylene
  • PS polystyrene
  • PE polyethylene
  • HDPE high density polyethylene
  • PC polycarbonate
  • PA polyacetate
  • PMMA poly-methyleene-methacrylate
  • PVDF poly-vinylidene-fluoride
  • the choice of material is not only governed by thermal and chemical considerations, but also economic considerations such as material costs, production technology etc.
  • One suitable method of production is injection moulding. Vacuum die-casting is another possible method of production.
  • the adapter is of course manufactured under conditions rendering it sterile and free from
  • the device comprises physical characteristics aiding in the correct positioning of the device in relation to surrounding devices and in relation to the receiving microtitre plate.
  • Such physical characteristic can comprise veits, grooves, colour codes and pins, with corresponding members in the adjacent device. In practise this can be realised by coloured fields, forming a pre-determined pattern when the devices are correctly assembled.
  • hte device is equipped with pins on at least one side, said pins fitting in corresponding holes on at least one side of the adjacent device, when the devices are correctly assembled.
  • velts or raised details can be provided, said velts or raised details fitting in corresponding grooves or depressions on the adjacent device when correctly assembled.
  • the device according to the invention comprises predispensed reagents, preferably in reagent cartridges or capillaries which are emptied by c ⁇ ntrifugation.
  • reagents can be single- or multi-compartment cartridges or single- or multi-lumen capillaries.
  • reagents can be added to the samples or reaction mixtures before or during the compression or transferral to a denser format. This is achieved through the fact, that reagent cartridges and capillaries can be made to release their content at predefined temperatures or centrifugal forces.
  • reagent cartridges see e.g. WO 98/10866.
  • a method according to the present invention includes both the possibility, that the adapter is delivered with suitable reagent cartridges or capillaries in place and the possibility, that these are added to the wells of the adapter after addition of the sample or reagent mixture that is to be compressed to a denser format.
  • the channels in the adapter, connecting the openings at the upper surface, corresponding to a less dense format, and the openings at the lower surface, corresponding to a more dens format are designed not to allow the passage of the reagent cartridge or the reagent capillary.
  • the devices according to the invention fits into a frame, said frame also being part of the invention.
  • the frame is designed to fit the receiving microtitre plate and to hold the devices in an orientation and position in relation to each other and in relation to the receiving microtitre plate as to guarantee flawless transfer of the sample or reagents from one format to the other.
  • the adapter and methods of its use, according to the present invention, offer the user numerous benefits.
  • the device functions as an interface between manual handling of the traditional 96-well format and the automated handling of the newer 384-well format and more dense formats.
  • the device makes it possible to transfer samples and reaction mixtures from one format into a more dense one with little or no risk for error, as the alignment of the corresponding wells is an automatic, "built-in" feature of the adapters.
  • the possibility of conducting an exactly determinable part of a reaction in the adapter, after which the reaction mixture is transferred into the receiving format offers many advantages.
  • the inventive device and method is superior, both in regard of the initial costs and sample economy.
  • the inventive device and method is particularly suited for supplying old samples, stored in serial format or on 96-well plates, to the newly developed methods and apparatuses used in high throughput screening and combinatorial chemistry.
  • the device and method of the present invention constitute a convenient interface between manual and automated operations in all types of small volume reactions, e.g. in fields like diagnostic analysis, biochemical analysis and high throughput screening and combinatorial chemistry.
  • Example 1 Four devices according to the invention are assembled in a frame, and placed over the first quadrant of a 16 by 24 microtitre plate. Samples are manually pipetted into the wells of the devices, now resembling a conventional 8 by 12 microtitre plate. This way the first 96-samples are compressed to the 384-well format. The process is repeated by first assembling four devices over the second, third and fourth quadrant of the receiving 16 by 24 plate.
  • Four 96-well microtitre plates are filled with samples and reagents in a conventional manner and subjected to temperation in a thermo shaker, built for this format.
  • Four devices according to the invention are assembled in a frame and placed over the first quadrant of a 16 by 24 microtitre plate.
  • the first 96-well plate is emptied in the assembled devices and the reaction mixtures thus transferred and compressed to the 384-well format
  • the process is repeated with new devices, assembled in lots of four, corresponding to the second, third and fourth quadrant of the receiving 16 by 24 plate.
  • Example 3 Four adapters assembled to form a 96-well grid on their combined upper surfaces, are supplied with reagent cartridges, for example single- or multilumen capillaries, containing one or several reagents.
  • the adapters are connected, up-side-down to a conventional 96-well microtitre plate, forming a tight seal between the openings of the wells on the 96-well microtitre plate and the openings of the adapter.
  • This combination is then turned and connected to the receiving, denser format, e.g. a 384-well microtitre plate and placed in a centrifuge.
  • the receiving, denser format e.g. a 384-well microtitre plate and placed in a centrifuge.
  • the reagent cartridge or single- or multilumen capillaries are emptied of their contents and at another, predetermined speed, the entire content of the former 96-well plate and the reagents are transferred into the receiving, 384-well plate.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

A l'aide d'un dispositif tel qu'un adaptateur on peut comprimer le format d'une plaque de microtitration classique à 96 cupules pour constituer un format à 384 cupules ou bien des formats encore plus denses. L'adaptateur peut également faire office de cuve à réaction dans une forme de réalisation dans laquelle il est vidé par centrifugation pour produire le format de réception plus dense. En outre, cet adaptateur peut comprendre des réactifs, qui se présentent de préférence sous forme de capillaires réactifs qui sont vidés lorsque la centrifugation a atteint un niveau prédéterminé.
PCT/SE2000/001632 1999-08-25 2000-08-24 Dispositif et procede permettant de manipuler et de traiter des echantillons et/ou des melanges reactionnels de petit volume WO2001014063A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP00957213A EP1214147A1 (fr) 1999-08-25 2000-08-24 Dispositif et procede permettant de manipuler et de traiter des echantillons et/ou des melanges reactionnels de petit volume
AU68866/00A AU6886600A (en) 1999-08-25 2000-08-24 Device and method for handling small volume samples and/or reaction mixtures
JP2001518192A JP2003531358A (ja) 1999-08-25 2000-08-24 小体積のサンプルおよび／または反応混合物を取り扱うデバイスおよび方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE9903002A SE9903002D0 (sv) 1999-08-25 1999-08-25 Device and method for handling small volume samples and/or reaction mixtures
SE9903002-5 1999-08-25

Publications (1)

Publication Number Publication Date
WO2001014063A1 true WO2001014063A1 (fr) 2001-03-01

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PCT/SE2000/001632 WO2001014063A1 (fr) 1999-08-25 2000-08-24 Dispositif et procede permettant de manipuler et de traiter des echantillons et/ou des melanges reactionnels de petit volume

Country Status (5)

Country Link
EP (1) EP1214147A1 (fr)
JP (1) JP2003531358A (fr)
AU (1) AU6886600A (fr)
SE (1) SE9903002D0 (fr)
WO (1) WO2001014063A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003055589A2 (fr) * 2001-12-31 2003-07-10 Institut für Physikalische Hochtechnologie e.V. Plaque de microtitration pour l'execution en parallele de plusieurs microsyntheses, en particulier a temperatures elevees
US8337775B2 (en) 2002-02-26 2012-12-25 Siemens Healthcare Diagnostics, Inc. Apparatus for precise transfer and manipulation of fluids by centrifugal and or capillary forces
WO2016139424A1 (fr) * 2015-03-03 2016-09-09 Nicolas Ugolin Electrodot : procede d'analyse d'echantillons biologiques en matrice de spots fixe par l'action d'un courant electrique conjugue a une difference de pression

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998024543A1 (fr) * 1996-12-02 1998-06-11 Glaxo Group Limited Procede et dispositif de transfert et de combinaison de reactifs
DE19712195A1 (de) * 1997-03-22 1998-09-24 Univ Schiller Jena Verfahren und Vorrichtung zum off-line Nachweis von Analyten nach MALDI-Massenspektrometrie
US5840256A (en) * 1996-04-09 1998-11-24 David Sarnoff Research Center Inc. Plate for reaction system
EP0955084A1 (fr) * 1998-04-27 1999-11-10 Corning Incorporated Réservoir capillaire refilé

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5840256A (en) * 1996-04-09 1998-11-24 David Sarnoff Research Center Inc. Plate for reaction system
WO1998024543A1 (fr) * 1996-12-02 1998-06-11 Glaxo Group Limited Procede et dispositif de transfert et de combinaison de reactifs
DE19712195A1 (de) * 1997-03-22 1998-09-24 Univ Schiller Jena Verfahren und Vorrichtung zum off-line Nachweis von Analyten nach MALDI-Massenspektrometrie
EP0955084A1 (fr) * 1998-04-27 1999-11-10 Corning Incorporated Réservoir capillaire refilé

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003055589A2 (fr) * 2001-12-31 2003-07-10 Institut für Physikalische Hochtechnologie e.V. Plaque de microtitration pour l'execution en parallele de plusieurs microsyntheses, en particulier a temperatures elevees
WO2003055589A3 (fr) * 2001-12-31 2003-11-06 Inst Physikalische Hochtech Ev Plaque de microtitration pour l'execution en parallele de plusieurs microsyntheses, en particulier a temperatures elevees
US8337775B2 (en) 2002-02-26 2012-12-25 Siemens Healthcare Diagnostics, Inc. Apparatus for precise transfer and manipulation of fluids by centrifugal and or capillary forces
WO2016139424A1 (fr) * 2015-03-03 2016-09-09 Nicolas Ugolin Electrodot : procede d'analyse d'echantillons biologiques en matrice de spots fixe par l'action d'un courant electrique conjugue a une difference de pression
FR3033409A1 (fr) * 2015-03-03 2016-09-09 Nicolas Ugolin Electrodot : procede d'analyse d'echantillons biologiques en matrice de spots fixes par l'action d'un courant electrique conjuguee a une difference de pression

Also Published As

Publication number Publication date
AU6886600A (en) 2001-03-19
SE9903002D0 (sv) 1999-08-25
JP2003531358A (ja) 2003-10-21
EP1214147A1 (fr) 2002-06-19

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