EP1239960B1 - Plaque a microtitration et procede de fabrication associe - Google Patents

Plaque a microtitration et procede de fabrication associe Download PDF

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Publication number
EP1239960B1
EP1239960B1 EP00988309A EP00988309A EP1239960B1 EP 1239960 B1 EP1239960 B1 EP 1239960B1 EP 00988309 A EP00988309 A EP 00988309A EP 00988309 A EP00988309 A EP 00988309A EP 1239960 B1 EP1239960 B1 EP 1239960B1
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EP
European Patent Office
Prior art keywords
filter means
wells
filter
spouts
opening
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EP00988309A
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German (de)
English (en)
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EP1239960A1 (fr
Inventor
Karl-Andreas Moll
Roland Beck
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3M Innovative Properties Co
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3M Innovative Properties Co
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Priority to EP00988309A priority Critical patent/EP1239960B1/fr
Publication of EP1239960A1 publication Critical patent/EP1239960A1/fr
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    • 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
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration

Definitions

  • the present invention relates to a method for producing a micro-titer test plate. Further, this invention relates to a particular micro-titer test plate that can be produced in connection with the method of the present invention.
  • Multi-well test plates also called micro-titer plates or micro-titer test plates
  • Micro-titer test plates have been described in numerous patents including US-A-4,948,442, US-A-3,540,856, US-A-3,540,857, US-A-3,540,858, US-A-4,304,865, US-A-4,948,564, US-A-5,620,663, US-A-5,464,541, US-A-5,264,184, WO 97/41955, WO 95/22406, EP-A-0 645 187 and EP-A-0 098 534.
  • Selected wells in the micro-titer test plate can be used to incubate respective microcultures or to separate biological or biochemical material followed by further processing to harvest the material.
  • Each well has filtration means so that, upon application of a vacuum to one side of the plate, fluid in each well is expressed through the filter leaving solids, such as bacteria and the like, entrapped in the well.
  • the filtration means can also act as a membrane such that certain materials in the test specimen are selectively bonded or otherwise retained in the filter means. The retained material may thereafter be harvested by means of a further solvent.
  • the liquid expressed from the individual wells through the filter means may be collected in a common collecting vessel in case the liquid is not needed for further processing or alternatively, the liquid from the individual wells may be collected in individual collecting containers as disclosed in US-A-5,464,541 and EP-A-0 098 534.
  • micro-titer plates have been used that conform to a standardized size of 85.47 by 127.76 mm having 12 rows of 8 wells each.
  • Many expensive automation equipment has been designed to this standard.
  • Such should preferably be accomplished in the most cost effective way and it has been proposed to retain approximately the size of the micro-titer plates yet increasing the number of wells therein. This would require minimal changes in the automation equipment.
  • micro-titer plate Various methods are known to produce a micro-titer plate. These methods are typically designed to produce the standard micro-titer plates having 96 wells. For example, such plates may be manufactured as multi-layer structures including a single sheet of filter material disposed to cover the bottom apertures of all the wells, the filtration material being bonded to the periphery of one or more of the well apertures. Such a structure may suffer from a problem called "cross-talk" by which fluid from adjacent wells mingles through for example capillary action, gravity or application of pressure.
  • a micro-titer, multi-layer plate includes a substantially rigid culture tray provided with wells having upstanding edges or rims bondeding the wider openings to the wells, and incubation is achieved while the culture tray is held "upside-down", i.e. the rims are disposed below the sheet.
  • a sheet of filter paper is placed over the top of a substantially rigid harvester tray having a like plurality of wells, each disposed and dimensioned to provide a tight push-fit with respect to the periphery of the rim of a corresponding well in the culture tray.
  • the latter US patent proposes a method of manufacturing in which the wells of a culture tray and harvester tray are welded together with there between a filter sheet which extends across the openings of the wells.
  • this method still does not completely solve the problem of cross talk.
  • welding of the wells may not be sufficient to avoid capillary action to cause mingling of fluids from adjacent wells.
  • this problem will be even more enhanced with micro-titer plates that have a high number of wells per unit area.
  • micro-titer plate by providing an array of wells connected to each other having opposite inlet and outlet openings, separately die cutting filter means conforming to the opening of the wells from a filter sheet and then inserting the filter means into the individual wells of the micro-titer plate.
  • This method would have the disadvantage of being difficult to automate as the handling of the individual filter means would be complicated and cumbersome thus requiring sophisticated and expensive equipment.
  • the degree of complexity and risk of failure during production would substantially increase when the amount of wells per area increases.
  • WO 98/55233 discloses the punching and inserting of filter discs from a filter sheet into a harvester plate having a conical drain funnel.
  • micro-titer plates it is desirable to find a further method for producing micro-titer plates, which method is preferably convenient, cost effective, capable of producing micro-titer plates that have a high number of wells per unit area and which micro-titer plates preferably have a reduced problem of cross-talk and good separation performance.
  • a micro-titer test plate having a plurality of sample containers connected to each other.
  • Each sample container has one or more side walls enclosing the interior of said sample container, a bottom wall with an outlet opening and an opposite upper end that is open and defines an inlet opening.
  • the micro-titer plate is produced from a first and a second part. The first part will have a plurality of wells connected to each other and the second part has a conforming number and arrangement of a plurality of spouts connected to each other.
  • Each of the wells of the first part has one or more side walls enclosing the interior of the wells and each of the wells has an upper end that is open and that will define the inlet opening of the sample containers and an opposite bottom opening. At their bottom opening, each of the wells will be bonded to the second part.
  • the wells will be tubular but they may also have a cross-section of a different shape parallel to the plane of the bottom openings. Further, the size of the cross-section in the axial direction of the wells may vary.
  • Each of the spouts of the second part encloses at its first end an opening that will define the outlet opening of a sample container once the two parts have been bonded together to form the micro-titer plate.
  • the first end of the spout will also define the bottom wall of the sample container.
  • the second end is defined by the free end of the spout.
  • the spouts may be provided at their first end with one or more walls enclosing an upper opening that is adapted for receiving the filter means. These walls extend in the axial direction away from the second end of the spouts.
  • the spouts taper towards their second end and they may be surrounded by a collar, co-axially extending from the first end.
  • the first and second part will generally be formed from a thermoplastic material and can be produced by injection molding.
  • thermoplastic materials that can be used include polystyrenes, polyvinyl chloride (including homo and copolymers thereof), polyethylenes and polyvinylidene chloride.
  • a filter sheet is placed on the side of the first part such that the filter sheet extends across each of the wells of the first part.
  • the sheet is placed on the side of the first part that has the bottom openings of the wells.
  • the filter sheet may be placed on this side of the second part and will then extend across each of the upper openings.
  • the filter sheet may be placed such as to directly overlay the openings of the wells or upper openings of the second part, but preferably, a die cut plate is provided between the filter sheet and the openings of the wells or the upper openings of the second part.
  • Such a die cut plate will have openings conforming to the shape and size of the desired filter means and the die cut plate will be placed in register with the openings of the wells or the upper openings of the second part.
  • a cutting stem may then penetrate the openings of the die cut plate thereby cutting the filter means out of the filter sheet.
  • the cutting stem may then also push the filter means in the openings of the wells or the upper openings of the second part.
  • the filter means When the filter means have been placed in the openings of the wells, the filter means will abut along their periphery the inner surface of the one or more side walls enclosing the interior of the wells.
  • the filter means When the filter means are placed in the upper openings of the second part, the filter means will preferably abut along their periphery the inner surface of the side walls enclosing the upper opening as well as the first end of the spouts. It is also possible to cut the filter means out of the filter sheet by means of other cutting techniques such as laser cutting and cutting by means of water jets or by providing sharp edges circumscribing the bottom opening of the wells of the first part or circumscribing the upper opening adapted for receiving filter means of the spouts of the second part.
  • other cutting techniques such as laser cutting and cutting by means of water jets or by providing sharp edges circumscribing the bottom opening of the wells of the first part or circumscribing the upper opening adapted for receiving filter means of the spouts of the second part.
  • a die cut plate will not be necessary and the filter means will be cut out while overlying the wells or spouts and they are thereafter pressed into the bottom openings of the wells or if provided, in the upper openings on the first end of the spouts.
  • filter means and “filter sheet” in connection with this invention are meant any means or sheet that can cause separation of one or more components from a mixture of components.
  • the terms “filter means” and “filter sheet” include sheets that can separate a solid component from the liquid in a dispersion as well as a membrane or sheet which can separate components which may be dissolved by selectively binding them.
  • the filter means of the present invention for example are means that allow selective adsorption, in particular of nucleic acids and proteins from liquids containing complete plant, animal or human cells or parts thereof.
  • the filter sheet and filter means in connection with the present invention may be single layer sheets or means but they are preferably laminates comprising several layers.
  • the filter sheet and filter means can be a laminate of a pre-filter layer, a solid phase extraction medium preferably in the form of a membrane and a porous support layer.
  • the filter means of the present invention will typically have a rigidity such that they will not substantially deform and substantially stay in place while being used so as to be capable of performing its separation function in the micro titer test plate.
  • the plurality of filter means will be preformed in the filter sheet according to feature (b) of claim 1.
  • preformed in the filter sheet is meant that the shape and size of the plurality of filter means is substantially formed in the filter sheet but wherein the filter means continue to be held within the filter sheet such that they do not accidentally separate from the filter sheet during its handling.
  • Preforming of the filter means can be carried out by partially cutting out the filter means from the filter sheet prior to placing the filter sheet on one side of the first or second part. Such partial cutting may be carried out by any cutting means known to those skilled in the art including, cutting by means of knifes, laser or water jets.
  • the filter means are cut out in such a way that the filter means stay connected to the filter sheet at one or more points on their periphery.
  • stay connected at one or more points on the periphery is meant that the major part of the periphery of the filter means is cut out and only a small portion on the periphery is not cut. At the minimum, the portion of the periphery that is not cut should be sufficient to retain the filter means in the filter sheet during further handling in the manufacturing of the micro-titer plate.
  • the filter sheet is a laminate of a prefilter layer and a porous support layer with a solid phase extraction medium there between.
  • the filter means can then be preformed in the filter sheet by ultrasonically welding the prefilter layer and the porous support layer together at the periphery of the filter means.
  • the prefilter layer and porous support layer are welded together at the complete periphery of the filter means.
  • the preformed filter means will then be comprised of the solid phase extraction medium that is enclosed by the prefilter layer and porous support layer that are welded together.
  • Such preformed filter means can be subsequently separated from the filter sheet when overlaying the array of sample containers by punching the preformed filter means out of the filter sheet without substantial dust formation.
  • dust formation during the separation of the filter means from the filter sheet may be further reduced by also partially cutting the preformed filter means at their periphery where the prefilter layer and support layer are welded together. The additional partial cutting can be carried out as described above.
  • the internal solid phase extraction medium can be in a variety of forms, such as fibers, particulate material, a membrane, other porous material having a high surface area, or combinations thereof.
  • the SPE medium is in the form of a membrane that includes a fibril matrix and sorptive particles enmeshed therein.
  • the fibril matrix is typically an open-structured entangled mass of microfibers.
  • the sorptive particles typically form the active material.
  • active it is meant that the material is capable of capturing an analyte of interest and holding it either by adsorption or absorption.
  • the fibril matrix itself may also form the active material, although typically it does not.
  • the fibril matrix may also include inactive particles such as glass beads or other materials for enhanced flow rates.
  • the prefilter layer is a porous material that can be made of a wide variety of materials. Typically, and preferably, it is made of a nonwoven material. More preferably, it is a nonwoven web of melt blown microfibers. Such "melt blown microfibers” or simply “blown microfibers” or “BMF” are discrete, fine, discontinuous fibers prepared by extruding fluid fiber-forming material through fine orifices in a die, directing the extruded material into a high-velocity gaseous stream to attenuate it, and then solidifying and collecting the mass of fibers.
  • the prefilter layer includes a nonwoven web of melt blown polyolefin fibers, particularly polypropylene fibers.
  • the prefilter layer preferably has the following characteristics: a solidity of no greater than about 20%; a thickness of at least about 0.5 millimeters (mm); and a basis weight of at least about 70 grams per square meter (g/m 2 ).
  • solidity refers to the amount of solid material in a given volume and is calculated by using the relationship between weight and thickness measurements of webs. That is, solidity equals the mass of a web divided by the polymer density divided by the volume of the web and is reported as a percentage of the volume.
  • the thickness refers to the dimension of the prefilter through which the sample of interest flows and is reported in mm.
  • the basis weight refers to mass of the material per unit area and is reported in g/m 2 .
  • the support layer can be made of a wide variety of porous materials that do not substantially hinder flow of the liquid of the sample of interest.
  • the porous material is typically a material that is capable of protecting the solid phase extraction medium from abrasion and wear during handling and use.
  • the material is sufficiently porous to allow the liquid sample to flow through it, although it does not allow particles that might be within the solid phase extraction medium from contaminating the sample.
  • the support layer is made of a nonwoven material.
  • the material of the prefilter and the support layer are very similar in composition (as opposed to structure), and more preferably, they are the same.
  • the plurality of preformed filter means conform in arrangement, number and shape to the arrangement, number and shape of the bottom openings of the wells or the optional upper openings on the second part.
  • the size of the filter means will typically be such that when the filter means are placed in the bottom openings of the wells or upper openings of the second part, the periphery of the filter means will abut the inner surface of the side walls of the wells or the inner surface of the side walls forming the upper openings.
  • the filter sheet will be placed in register with the bottom openings of the wells or the upper openings of the second part and the filter means can then be separated from the filter sheet and inserted in the openings.
  • Separation of the filter means can be caused by pressing the filter means in the sample container thereby tearing off the filter means from the remainder of the filter sheet or alternatively, the preformed filter means may be separated by cutting and subsequently or simultaneously pressing the filter means into the bottom openings of the wells or the upper openings on the second part.
  • the remainder of the filter sheet from which the filter means have been separated is removed and the first and second part are then bonded together.
  • Bonding the two parts together is carried out by bringing the first and second part together such that the bottom openings of the wells face the upper first end of the spouts of the second part. Both parts are bonded together by bonding each of the wells of the first part to each of the spouts of the second part in an irreversible and permanent way.
  • irreversible and permanent is meant that the two parts can no longer be separated from each other without damaging the micro-titer plate.
  • Bonding the two parts together is further accomplished in such a way that each of the plurality of formed sample containers connected to each other, are each sealed with respect to each other.
  • a preferred means for binding the wells to the spouts includes thermal bonding and in particular ultrasonic welding.
  • the bottom opening of the wells of the first part may be circumscribed with one of a groove and ridge. The first end of the spouts will then be provided with the other of the groove and ridge.
  • the wells may be bonded to the spouts by mutually engaging mechanically means that snap into each other such that they cannot be disengaged without damaging the micro-titer plate formed.
  • the wells can be glued to the spouts or they may be molded to the spouts. In the latter case, after the first and second part have been engaged with each other, a small opening would remain that circumscribes each of the sample containers near the interface between the spouts and the wells. This opening is then subsequently filled with a thermoplastic polymer via an injection molding.
  • a micro-titer plate comprising sample containers connected to each other is obtained.
  • Each of the sample containers formed has one or more side walls enclosing the interior of the sample container, an upper end that is open and defines an inlet opening and an opposite bottom wall that has an outlet opening which is enclosed by a spout.
  • the bottom wall of the sample container with the outlet opening is formed by the first end of the spouts and the inlet opening is formed by the open upper end of the wells of the first part.
  • the filter means abuts the bottom wall and abuts along its periphery the inner surface of the one or more side walls that enclose the interior of the sample container.
  • the sample containers may further contain a band enclosing an opening.
  • This band can be inserted in each of the sample containers to press the filter means against the bottom wall of the sample container.
  • the band abuts along its periphery, the inner surface of the side wall(s) of the sample containers.
  • the bands generally conform to the shape of the sample container and are preferably rings when the sample containers are tubular.
  • the bands are preferably plastic or rubbery.
  • the wall(s) of the wells of the first part may be provided thinner for a portion proximate to the bottom opening of the well so as to adapt the bottom opening for receiving a filter means.
  • the wall(s) of the wells may be thickened over a portion at a certain distance away from the bottom opening.
  • the distance away from the bottom opening will generally be chosen such as to adapt the bottom opening for receiving a filter means such that the filter means will be pressed against the bottom of the sample container when the first and second part are bonded together.
  • the second part may have at the first end of the spouts, upper openings adapted for receiving a filter means.
  • the filter means will be cut out and placed in the upper openings of the second part such that they abut the first end of the spouts and the inner surface of the wall or walls defining the upper opening. If the size of the upper openings is elected to be somewhat larger than the size of the bottom opening of the wells of the first part, then when the first and second part are bonded together, the wails of the wells of the first part will press the filter means against the bottom wall in the sample containers of the micro-titer plate so produced.
  • a micro-titer plate in which a band or similar means is not necessary to press the filter means against the bottom wall of the sample containers.
  • a micro-titer plate according to this aspect of the invention comprises a plurality of sample containers connected to each other, each sample container having one or more side walls enclosing the interior of the sample container, an open upper wall defining an inlet opening and a bottom wall having an opening defining an outlet opening, the outlet opening connecting to a spout extending in the axial direction of the sample container, wherein the sample container contains a filter means that is in abutment with the bottom wall and side walls of the sample container and wherein one or more of the side walls of the sample container are adapted to press the filter means to the bottom walls.
  • Micro-titer plates produced in accordance with the present invention generally are less prone to cross-talk, are fairly convenient to produce, and have a good separation performance.
  • the micro-titer plates of the present invention it is possible to perform a physical separation, a chemical separation, or a bio-polymer separation or extraction of liquids containing plant, animal or human cells, and it allows, in particular, to perform the separation of nucleic acids and/or proteins of the cells.
  • the liquid in the sample container penetrates a filter means having selectively adsorbing material, the liquid leaving the filter means and entering a collecting container.
  • the filter means having selectively adsorbing material has chromatographic properties, which can include ion exchange properties or affinity-chromatographic properties, if the filter means comprises suitable affinity ligands.
  • a preferred filter means comprises a fibrillated polytetrafluoroethylene matrix having enmeshed therein sorptive derivatized silica particulate as are disclosed in US-A-4,810,381 and US-A-4,699,717, respectively. Subsequently, the collecting container is replaced by another one, and a liquid containing a solvent is applied on the filter means, which selectively removes a certain portion of the material adsorbed in the filter means so that it may enter the collecting container.
  • the filter means of the device of the present invention may comprise one or several layers.
  • Preferred filter means comprise a fibrillated polytetrafluoroethylene matrix having sorptive particulate enmeshed therein, as is disclosed, for example, in US-A-4,810,381.
  • the filter means may be formed by two porous fixation means, in particular frits, with particles therebetween.
  • the particles can be in the form of bulk material, have chromatographic properties as described before.
  • the preferred particles are made from a material that is based on silica gel, dextran or agarose. Frits may consist of glass, polyethylene (PE) or polytetrafluoroethylene (PTFE) and have a pore size of about 0.1-250 ⁇ m, preferably about 100 ⁇ m.
  • the thickness of the particle layer is about 1-10 mm, preferably 2.5 mm, with a particle size of 1-300 ⁇ m, preferably 16-23 ⁇ m.
  • the filter means has a support membrane in which the adsorptive particles are embedded. Since the support membrane can be rather weak and there being a possibility that it can burst when a partial vacuum is applied on it (of comparatively high pressure difference), a back-up fabric or fibrous layer can be arranged below the support membrane, which provides integrity to the support membrane on the bottom wall of the sample container and preferably consists of a non-woven polyalkylene fibrous material such as polypropylene or polyethylene.
  • the micro-titer plate of the present invention is not limited to the dimensions of the single parts mentioned herein. Generally, the micro-titer plate of the invention can be produced in any desired size. Nevertheless, the method of the present invention is particularly suitable for producing micro-titer plates that have a large number of sample containers per unit of area without a substantial risk of cross-talk. For example the method of the present invention can be used to make a micro-titer plate having a length between 11 and 13 cm and a width between 8 and 9 cm and having from 90 to 400 sample containers. For example, a micro-titer plate of the aforementioned dimensions and having 96 or 384 sample containers may be produced with the method of the present invention.
  • Another aspect of the present invention is a filter sheet comprising a plurality of preformed filter means according to claim 9.
  • FIG. 1 there is shown a three dimensional view of a micro-titer test plate that can be produced with the method of the present invention.
  • the micro-titer test plate has a plurality of sample containers 10 connected to each other. As shown in Fig. 1, the sample containers are connected at the inlet openings 14 with each other by a plate 72 and near the spouts 24 by the plate 42.
  • the micro-titer test plate of Fig. 1 is produced using a first and second part that will permanently and irreversibly be bonded to each other according to the method of the invention.
  • Figs. 2a and 2b show a first embodiment of the manufacturing method of the invention.
  • Fig. 2a there is shown a partial cross-section of a first part 60 having a plurality of wells 30 connected to each other.
  • Each of the wells 30 has side walls 31 enclosing the interior of the wells.
  • Each of the wells 30 has an opening which will form the inlet opening 14 of the sample container and an opposite bottom opening 16.
  • the bottom opening 16 is adapted for receiving a filter means by providing thickened portions 33 having a bottom wall portion 33' on the side walls 31.
  • the thickened portions 33 are provided at a predetermined distance 12 from the bottom opening.
  • This predetermined distance 12 will generally be selected such that when the filter means 39 have been placed therein and the first part 60 has been bonded to the second part 80, the filter means 39 will be pressed against the first end 23 of the second part 80 which will form the bottom wall of the sample containers (see Figs. 2c and 2d).
  • predetermined distance 12 will correspond to or be somewhat less than the thickness of the filter means 39.
  • the thickened portion 33 of the side walls near the bottom opening may be provided by gradually thickening the side walls towards the interior of the well from a first point downwards towards the bottom opening 16 and then at a second point abruptly reducing the thickness of the side walls, preferably to the thickness of the side walls at the first point.
  • the side walls may have a first thickness from the inlet opening 14 towards the bottom opening and this first thickness may be reduced to a second thickness over a predetermined distance 12 at the bottom opening so as to adapt that bottom opening for receiving a filter means.
  • This embodiment is shown in Fig. 4a and 4b.
  • a die cut plate 100 that has openings 102 that conform in shape and size to the bottom opening 16 of the wells of the first part 60.
  • the die cut plate further has grooves 101 that mate with corresponding ridges 36 that circumscribe the bottom opening 16.
  • Die cut plate 100 is engaged with first part 60 whereby the ridges 36 mate with grooves 101.
  • a filter sheet 1 is provided on the die cut plate and filter means 39 are cut out of the filter sheet 1 by a plurality of cutting stems 90 which also force the filter means 39 into the bottom opening 16 adapted for the receiving means (see Fig. 2b).
  • the plurality of cutting stems 90 are then removed as well as the remainder of the filter sheet 1 and the die cut plate 100.
  • the remainder of the filter sheet 1 will be a sheet with holes corresponding to the filter means 39 that have been cut out.
  • a second part 80 (see Fig. 2c) is then provided that has a plurality of spouts 24 connected to each other that enclose an opening 22 at a first end 23.
  • the first end 23 will form the bottom wall of the sample containers 10 (see Fig. 1) and the opening 22 will define the outlet opening of the sample containers 10.
  • Opposite the first end is the second end 81 of the spouts 24.
  • spouts 24 taper towards the second end 81.
  • the spouts 24 are connected to each other via a plate 42. Grooves 35 are provided and circumscribe the first end 23 of the spouts 24.
  • the first part with the filter means 39 inserted in the bottom openings 16 of the wells is placed on the second part 80 such that the bottom openings 16 face the first ends of the spouts.
  • Fig. 2d shows that ridges 36 are inserted in the grooves 35 when the first part 60 and second part 80 are contacted with each other and are bonded with the corresponding spouts on the second part 80 by ultrasonically melting the ridges 36 with the grooves 35. Accordingly, a micro-titer plate is then obtained in which the sample containers are sealed relative to each other such that there is little or no potential for cross-talk.
  • a second part 50 having a plurality of spouts 24 connected to each other is provided.
  • Spouts 24 each enclose an opening 22 at first ends 23. Opposite to the first end of the spouts is the second end 81. Spouts 24 taper towards second end 81.
  • the spouts 24 are connected to each other by plate 42.
  • Each of the spouts 24 of the second part 50 also has side walls 51 circumscribing the first end 23 and defining an upper opening 55 adapted for receiving filter means 39.
  • the height of the side walls 51 will generally be elected such that when the second part 50 is bonded with a first part 110, the side walls of the wells of that first part 110 may press the filter means 39 against the first ends of the spouts 24 (see Fig. 3d) forming the bottom walls of the sample containers 10. Typically therefore, the height will be equal to or somewhat less than the thickness of the filter means 39.
  • a die cut plate 100 having grooves 101 capable of mating with ridges 52 provided on the walls 51 and circumscribing the upper openings 55 is provided.
  • the die cut plate 100 is contacted with the second part such that grooves 101 mate with ridges 52.
  • a filter sheet 1 is placed thereon such that it extends across each of the upper openings 55 of the second part.
  • Cutting stems 90 then cut out the filter means 39 and press them into the upper openings 55 (Fig. 3b).
  • the die cut plate 100 and remainder of the filter sheet 1 are then removed and a first part 110 for bonding with the second part 50 is provided.
  • the first part 110 as shown in Fig. 3c has a plurality of wells 115 connected to each other, each of which has side walls 113 defining the interior of the wells.
  • Each of wells 115 further has at one end an opening defining an inlet opening 114 and at the opposite end an opening defining the bottom opening 116.
  • Side walls 113 are gradually thickened at portion 117 near the bottom opening 116 such that when the first part 110 is bonded with the second part 50, the bottom wall portion 113' of side walls 113 will partially overlap with the filter means 39 so as to press the latter against first ends 23 of the spouts of the second part 50.
  • the bottom wall portion 113' of side walls 113 are further provided with grooves 111 that circumscribe the bottom opening 16 and into which ridges 52 of the second part 50 can be inserted. Ultrasonic bonding or other thermal bonding techniques may thus permanently and irreversibly bind each of the wells of first part 110 to the corresponding spouts of second part 50 by melting ridges 52 with grooves 111.
  • a further embodiment of the present invention in which the first and second part are permanently and irreversibly bonded together by mutually engaging mechanical means.
  • a first part 120 have a plurality of wells 125 connected to each other that each have side walls 121 enclosing the interior of the wells.
  • an opening defining inlet opening 124 and opposite thereof is bottom opening 126.
  • the bottom opening 126 is adapted for receiving filter means 39 by providing thickened portion 123 having bottom wall portion 123' proximate to the bottom opening 126.
  • each of the wells 125 of the first part 120 are provided with the female portion 128 of mutually engaging and interlocking mechanical means.
  • Female portion 128 has snap-in holes 129 from which the corresponding male heads 203 (see 6c) cannot be withdrawn once they have been snapped into holes 129.
  • Female portion 128 further has sharp edges that are capable of cutting through at least some filter sheets.
  • a filter sheet 1 is provided on the side of the first part 120 that has the bottom openings and female portions 128 with sharp edges 127.
  • Filter sheet 1 is provided between first part 120 and a plate 105.
  • the first part 120 is pressed onto the filter sheet 1 thereby cutting out filter means 39 and simultaneously inserting them into the bottom openings 126. As can be appreciated from Fig. 6b, this also results in remaining portions 45 of the filter sheet 1 to be pressed into the female portions 128. These remaining portions 45 of the filter sheet 1 can be removed from the female portions 128 by applying a vacuum to plate 105 through channels 46.
  • the remaining portions 45 of the filter sheet 1 will be removed from the female portions 128.
  • the remaining portions 45 form a sheet with holes that correspond in shape, size and number to the filter means 39 that have been removed therefrom. As shown in Fig. 7, these holes are circular but they could be of a different form depending on the form of the bottom openings 126.
  • Fig. 6c shows the first part 120 with the filter means 39 inserted in bottom openings 126 and wherein the remaining portions 45 of the filter sheet 1 have been removed. Further shown is second part 200 for bonding with first part 120 to produce the micro-titer plate. Second part 200 has a plurality of spouts 24 connected to each other. Each spout 24 has a first end 201 that will ultimately form the bottom wall of the sample containers. The first end 201 has an outlet opening 202 that is enclosed by the spout. First ends 201 are each circumscribed by male portions 204 for engagement with female portions 128 of first part 120.
  • first part 120 and second part 200 of the micro-titer plate can no longer be separated from each other because the heads 203 irreversibly lock into holes 129.
  • the cutting of the filter means out of the filter sheet 1 is carried out by using a die cutting plate 100 and cutting stems 90 or is carried out by the sharp edges 127 on the first part 120 as shown in Figs. 6a-6d.
  • the filter sheet may contain the filter means 39 preformed therein by partially cutting them out.
  • filter means 39 are partially cut out in the filter sheet 5.
  • a plurality of filter means 39 are partially cut out which conform in arrangement, shape and number to the plurality of bottom openings of the wells of the first part or the upper openings of the second part in which they will be inserted.
  • Fig. 5 shows a few of such filter means 39 partially cut out in the filter sheet 5.
  • Filter means 39 of Fig. 5 have a circular periphery to conform to a circular bottom opening of the first part or upper opening of the second part. As can be seen, filter means 39 in Fig. 5 have been cut along there periphery except for two oppositely laying points 2, 3 where the filter means 39 remain connected to the filter sheet 5. While Fig. 5 illustrates filter means 39 as circular, filter means 39 may also have a square periphery to conform to openings that have a square cross-section. If a filter sheet 5 with the filter means 39 partially cut out is used instead of filter sheet 1, it will generally not be necessary to use a die cut plate.
  • the filter sheet 5 will be placed on the first or second part such that the filter means 39 are in register with the openings in which they are to be placed. They can then be separated from the filter sheet by means of a plurality of stems that push the filter means 39 into the relevant opening while at the same time tearing of the filter means at the points where they were still connected to the filter sheet.
  • This embodiment has the advantage that less dust is created when the filter means are separated from the filter sheet and accordingly there will be less risk that dust may interfere with the filter performance of the individual sample containers of the micro-titer plate produced.
  • the use of a filter sheet 5 with filter means 39 partially cut out further presents the advantage that when one of the first or second part has been equipped with sharp edges to cut the filter sheet, cutting will be facilitated.

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  • Health & Medical Sciences (AREA)
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  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Devices For Use In Laboratory Experiments (AREA)
  • Secondary Cells (AREA)
  • Glass Compositions (AREA)
  • Polishing Bodies And Polishing Tools (AREA)
  • Laminated Bodies (AREA)

Claims (9)

  1. Méthode de fabrication d'une plaque à essai par microtitration comprenant une pluralité de récipients d'échantillons raccordés les uns aux autres, dont chaque récipient d'échantillons comporte une ou plusieurs parois latérales entourant l'intérieur dudit récipient d'échantillons, une paroi inférieure avec une ouverture de sortie et une extrémité supérieure opposée qui est ouverte et définit une ouverture d'entrée, ladite méthode comprenant les étapes qui consistent à :
    (a) fournir une première partie et une deuxième partie, ladite première partie comprenant une pluralité de puits raccordés les uns aux autres et comportant chacun une extrémité supérieure qui est ouverte et qui définira l'ouverture d'entrée desdits récipients d'échantillons et une ouverture inférieure opposée, et ladite deuxième partie comprenant une pluralité de becs raccordés les uns aux autres, lesdits becs correspondant, par leur disposition et par leur nombre, auxdits puits de ladite première partie et chaque bec entourant, à une première extrémité, une ouverture qui définira l'ouverture de sortie desdits récipients d'échantillons, et chaque bec comportant éventuellement une ouverture supérieure à ladite première extrémité pour recevoir un moyen de filtration ;
    (b) fournir une feuille filtrante dans laquelle est préformée une pluralité de moyens de filtration, les moyens de filtration préformés constituant ladite pluralité correspondant, par leur disposition, leur nombre et leur forme, à la disposition, au nombre et à la forme de ladite pluralité de récipients d'échantillons et ladite pluralité de moyens de filtration étant préformée dans ladite feuille filtrante par soudage par ultrasons à la périphérie des moyens de filtration et/ou par découpage partiel des moyens de filtration, de sorte que chaque moyen de filtration reste raccordé à la feuille filtrante en au moins un point sur sa périphérie ;
    (c) placer une feuille filtrante qui s'étend en travers de chacun desdits puits de ladite première partie en correspondance exacte avec ceux-ci sur un côté de ladite première partie, ou placer une feuille filtrante qui s'étend en travers de chacun desdits becs de ladite deuxième partie en correspondance exacte avec ceux-ci sur le côté de ladite deuxième partie comportant lesdites premières extrémités ;
    (d) séparer de ladite feuille filtrante des moyens de filtration qui correspondent par leur forme, leur taille, leur disposition et leur nombre, soit à l'ouverture inférieure des puits de ladite première partie, soit, lorsque celles-ci sont présentes, aux ouvertures supérieures facultatives présentes à la première extrémité des becs ;
    (e) placer lesdits moyens de filtration dans chacune des ouvertures inférieures des puits ou, lorsque celles-ci sont présentes, dans chacune desdites ouvertures supérieures présentes à la première extrémité des becs ;
    (f) retirer le reste de ladite feuille filtrante de laquelle les moyens de filtration ont été séparés ;
    (g) mettre ladite première partie et ladite deuxième partie en contact l'une avec l'autre, de sorte que l'ouverture inférieure desdits puits se trouve face à la première extrémité desdits becs ; et
    (h) lier ladite première partie et ladite deuxième partie l'une à l'autre en liant à titre permanent et de façon irréversible chacun des puits de ladite première partie à chacun des becs de ladite deuxième partie, en formant ainsi une pluralité de récipients d'échantillons raccordés les uns aux autres, qui sont étanchés les uns par rapport aux autres.
  2. Méthode selon la revendication 1, dans laquelle on sépare lesdits moyens de filtration de ladite feuille filtrante en appuyant sur lesdits moyens de filtration pour les enfoncer dans l'ouverture inférieure de chacun des puits ou dans l'ouverture supérieure présente à la première extrémité de chacun des becs de ladite deuxième partie, en déchirant ainsi les moyens de filtration pour les séparer de ladite feuille filtrante.
  3. Méthode selon la revendication 1, dans laquelle lesdits becs comportent une ouverture supérieure à ladite première extrémité pour recevoir un moyen de filtration et l'ouverture inférieure desdits puits a une taille inférieure à celle du moyen de filtration, de sorte que quand ladite première partie et ladite deuxième partie ont été liées l'une à l'autre, la partie des parois latérales des puits qui constitue la paroi inférieure presse lesdits moyens de filtration contre la paroi inférieure desdits récipients d'échantillons.
  4. Méthode selon la revendication 1, dans laquelle on sépare lesdits moyens de filtration de ladite feuille filtrante en découpant lesdits moyens de filtration.
  5. Méthode selon la revendication 1, dans laquelle l'ouverture inférieure desdits puits a été pourvue de bords coupants et dans laquelle on sépare lesdits moyens de filtration en appuyant sur ladite première partie pour l'enfoncer dans ladite feuille filtrante.
  6. Méthode selon l'une quelconque des revendications précédentes, dans laquelle ladite première partie et ladite deuxième partie sont liées à titre permanent et de façon irréversible l'une à l'autre par thermocollage, collage, moulage des puits de ladite première partie sur lesdits becs de ladite deuxième partie, ou par un moyen à engagement mutuel présent sur lesdits puits de ladite première partie et lesdits becs de ladite deuxième partie.
  7. Méthode selon la revendication 6, dans laquelle ledit thermocollage est le soudage par ultrasons.
  8. Méthode selon l'une quelconque des revendications précédentes, dans laquelle chaque bec de ladite pluralité de becs s'amincit au fur et à mesure qu'on se rapproche de sa deuxième extrémité opposée à ladite première extrémité.
  9. Feuille filtrante comprenant une pluralité de moyens de filtration préformés, les moyens de filtration préformés constituant ladite pluralité correspondant, par leur disposition, leur nombre et leur forme, à la disposition, au nombre et à la forme des récipients d'échantillons d'une plaque à essai par microtitration et ladite pluralité de moyens de filtration étant préformée dans ladite feuille filtrante par soudage par ultrasons à la périphérie des moyens de filtration et/ou par découpage partiel des moyens de filtration, de sorte que chaque moyen de filtration reste raccordé à la feuille filtrante en au moins un point sur sa périphérie.
EP00988309A 1999-12-23 2000-12-21 Plaque a microtitration et procede de fabrication associe Expired - Lifetime EP1239960B1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP00988309A EP1239960B1 (fr) 1999-12-23 2000-12-21 Plaque a microtitration et procede de fabrication associe

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP99204505 1999-12-23
EP99204505A EP1110611A1 (fr) 1999-12-23 1999-12-23 Plaque de microtitrage et sa méthode de fabrication
PCT/US2000/035092 WO2001045844A1 (fr) 1999-12-23 2000-12-21 Plaque a microtitration et procede de fabrication associe
EP00988309A EP1239960B1 (fr) 1999-12-23 2000-12-21 Plaque a microtitration et procede de fabrication associe

Publications (2)

Publication Number Publication Date
EP1239960A1 EP1239960A1 (fr) 2002-09-18
EP1239960B1 true EP1239960B1 (fr) 2006-05-31

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Application Number Title Priority Date Filing Date
EP99204505A Withdrawn EP1110611A1 (fr) 1999-12-23 1999-12-23 Plaque de microtitrage et sa méthode de fabrication
EP00988309A Expired - Lifetime EP1239960B1 (fr) 1999-12-23 2000-12-21 Plaque a microtitration et procede de fabrication associe

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP99204505A Withdrawn EP1110611A1 (fr) 1999-12-23 1999-12-23 Plaque de microtitrage et sa méthode de fabrication

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EP (2) EP1110611A1 (fr)
JP (1) JP4602623B2 (fr)
AT (1) ATE327829T1 (fr)
AU (1) AU2453101A (fr)
DE (1) DE60028413T2 (fr)
WO (1) WO2001045844A1 (fr)

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CA2455345C (fr) 2001-05-31 2009-08-25 Pall Corporation Puits servant a traiter un fluide
US7122155B2 (en) 2002-07-16 2006-10-17 Mcgill University Electron microscopy cell fraction sample preparation robot
EP1587624B1 (fr) 2003-01-17 2011-12-14 Greiner Bio-One GmbH Recipient a echantillon pour analyses
US9005549B2 (en) 2003-01-17 2015-04-14 Greiner Bio-One Gmbh High throughput polymer-based microarray slide
DE10321042B4 (de) * 2003-01-17 2006-09-21 Greiner Bio-One Gmbh Biochip-Träger
US7063216B2 (en) 2003-09-04 2006-06-20 Millipore Corporation Underdrain useful in the construction of a filtration device
US8753588B2 (en) 2003-10-15 2014-06-17 Emd Millipore Corporation Support and stand-off ribs for underdrain for multi-well device
US20050236318A1 (en) * 2004-04-23 2005-10-27 Millipore Corporation Low holdup volume multiwell plate
US7618592B2 (en) 2004-06-24 2009-11-17 Millipore Corporation Detachable engageable microarray plate liner
FR2887159B1 (fr) * 2005-06-20 2007-08-17 Ct Hospitalier Regional De Nan Puits filtrant d'analyse biologique
JP5175599B2 (ja) * 2008-04-09 2013-04-03 富士紡ホールディングス株式会社 固相抽出用カートリッジ充填カラムの製造方法
WO2015016315A1 (fr) * 2013-08-02 2015-02-05 株式会社ニコン Plaque, procédé de production de plaque, procédé d'observation de biopuce et procédé de criblage
US10324089B2 (en) * 2014-12-11 2019-06-18 Critical Care Diagnostics, Inc. Test apparatus and methods for ST2 cardiac biomarker

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EP0009320B1 (fr) * 1978-08-31 1982-04-21 National Research Development Corporation Appareil et procédé pour récolter de la matière de plaques de micro-culture
US4948442A (en) * 1985-06-18 1990-08-14 Polyfiltronics, Inc. Method of making a multiwell test plate
US5108704A (en) * 1988-09-16 1992-04-28 W. R. Grace & Co.-Conn. Microfiltration apparatus with radially spaced nozzles
US5264184A (en) * 1991-03-19 1993-11-23 Minnesota Mining And Manufacturing Company Device and a method for separating liquid samples
SE9400436D0 (sv) * 1994-02-10 1994-02-10 Pharmacia Lkb Biotech Sätt att tillverka filterbrunnar
US6391241B1 (en) * 1997-06-06 2002-05-21 Corning Incorporated Method of manufacture for a multiwell plate and/or filter plate
JPH11276912A (ja) * 1998-03-27 1999-10-12 Mitsubishi Chemical Engineering Corp ガラス器具およびガラスフィルター並びにこれらの製造方法
US6159368A (en) * 1998-10-29 2000-12-12 The Perkin-Elmer Corporation Multi-well microfiltration apparatus
EP1110610A1 (fr) * 1999-12-23 2001-06-27 3M Innovative Properties Company Plaque de microtitrage avec des filtres insérés et procédé pour sa production

Also Published As

Publication number Publication date
EP1110611A1 (fr) 2001-06-27
ATE327829T1 (de) 2006-06-15
JP4602623B2 (ja) 2010-12-22
WO2001045844A1 (fr) 2001-06-28
JP2003518248A (ja) 2003-06-03
DE60028413T2 (de) 2006-11-30
DE60028413D1 (de) 2006-07-06
EP1239960A1 (fr) 2002-09-18
AU2453101A (en) 2001-07-03

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