EP1341611B1 - Porte-echantillon, son couvercle et procede de preparation d'analyse - Google Patents

Porte-echantillon, son couvercle et procede de preparation d'analyse Download PDF

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Publication number
EP1341611B1
EP1341611B1 EP01997350A EP01997350A EP1341611B1 EP 1341611 B1 EP1341611 B1 EP 1341611B1 EP 01997350 A EP01997350 A EP 01997350A EP 01997350 A EP01997350 A EP 01997350A EP 1341611 B1 EP1341611 B1 EP 1341611B1
Authority
EP
European Patent Office
Prior art keywords
sample carrier
cover
cover plate
liquid
sample
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.)
Expired - Lifetime
Application number
EP01997350A
Other languages
German (de)
English (en)
Other versions
EP1341611A1 (fr
Inventor
Nico Michaelsen
Martin Klumpp
Olaf Sollböhmer
Sabine KÖSTER
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.)
Evotec OAI AG
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Evotec OAI AG
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Filing date
Publication date
Application filed by Evotec OAI AG filed Critical Evotec OAI AG
Publication of EP1341611A1 publication Critical patent/EP1341611A1/fr
Application granted granted Critical
Publication of EP1341611B1 publication Critical patent/EP1341611B1/fr
Anticipated expiration legal-status Critical
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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
    • B01L3/508—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
    • B01L3/50853—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 with covers or lids

Definitions

  • the invention relates to a sample carrier, in particular a titer plate with a plurality of wells, can be introduced into the smallest samples. Furthermore, the invention relates to a lid for sample carrier and a method for examination preparation.
  • the smallest samples are, in particular, liquids or other samples having a volume of 0.01-50 ⁇ l, in particular 0.2-5 ⁇ l. Likewise, the samples may be cell culturing on this scale, preferably on a 1-5 ⁇ l scale.
  • the sample carriers may be chips.
  • chips usually have one or more channels, which may be, for example, capillary channels and possibly one or more liquid reservoirs.
  • it may be microfluidic chips with preferably two reservoirs, which communicate with each other via a channel are connected.
  • a fluid exchange takes place between the two reservoirs, which can be regulated by suitable valves, membranes, osmotic barriers and / or ion barriers.
  • the mixing behavior of liquids for example under the influence of electromagnetic forces can be investigated.
  • sample components can be separated from each other.
  • Known chips are made of silicon, glass, plastic or ceramic.
  • sample carriers can also be, for example, planar plates with a hydrophobic grid. Such plates are partially coated with a hydrophobic substance, so that droplets of sample liquid form in these areas, which can then be examined. Sample carriers may also be so-called SBS microtiter plate formats and the so-called Terasaki format.
  • the wells of titer plates can be produced, for example, by etching-milling or by using injection molding and laser machining.
  • a lid By providing a lid by which, for example, the pre-wells are sealed tightly in a titer plate, a corresponding sterility can be ensured and any evaporation from the wells of the titer plate can be prevented, but it can not set the desired cultivation conditions. For example, a CO 2 content between 5 - 7% for optimal cell cultivation (pH regulation of the cell culture medium) is needed, which can not be adjusted by the conventional arrangements.
  • WO 95/97196 a sample carrier is known whose individual wells can be sealed separately with the aid of a lid in order to avoid cross-contamination of adjacent wells.
  • the object of the invention is to provide a sample carrier, in particular a titer plate, a lid for a sample carrier and a method for preparation of investigations, in particular of chemical and / or biological micro samples, in which or in which, in particular, evaporation of samples is substantially avoided and at the same time optimal cultivation conditions, ie sufficient sterility and precise control of the gas exchange, in particular a precise classification of the CO 2 content for cells in sample carriers for very small samples, can be achieved.
  • the sample carrier has a receiving part for receiving small samples. If the sample carrier is a titer plate, the smallest samples are placed in wells of the titer plate. Is it the sample carrier around a chip, the smallest samples are introduced into channels or in depressions of the chip. Correspondingly, the samples are applied to the grid in a sample carrier designed as a planar plate with a hydrophobic grid, so that drops of sample material are formed.
  • the wells of a titer plate or the wells of a chip preferably have a volume of 0.01 to 50 ul. In particular, the volume is 0.2 to 5 ⁇ l.
  • the sample carrier has a lid, through which the receiving part is closed in such a way that a cavity is formed between a sample surface and the lid. The lid underside thus has a distance to the sample surface and generally also to the top of the receiving part.
  • the lid is designed in such a way that a saturated atmosphere in the cavity is essentially preserved under changing environmental conditions.
  • the lid thus has a certain vapor permeability and / or moisture absorption and / or dispensing capability.
  • the cover is designed such that it is suitable for receiving and dispensing liquid.
  • the saturated atmosphere in the cavity remains so constant under changing environmental conditions that there is a deviation of less than 5% from a saturation level. In particular, this deviation is less than 2%.
  • the cultivation conditions for z. B. provided in wells of a titer plate cells are maintained even in changing outdoor conditions.
  • the lid of the sample carrier according to the invention has a porous cover plate.
  • This is preferably made of ceramic or glass and serves as a liquid storage.
  • the resulting from the preferably stored in the porous cover plate liquid vapor phase creates in the cavity between the sample carrier and lid a saturated atmosphere, whereby the evaporation of sample liquid is prevented.
  • a gas exchange through the porous lid is possible, so that, for example, for cell cultivation, the optimal CO 2 concentration can be adjusted in the cavity.
  • the gas exchange takes place, for example, via pores not closed by wetting liquid, so that the gas can pass through the pores to the outside.
  • an indirect gas exchange by the gas, in particular CO 2 is dissolved in the liquid and ggl. out of the liquid into the cavity ausast.
  • the lid according to the invention is designed so that liquid in the form of vapor can escape from the pores of the lid material.
  • the sample carrier according to the invention is introduced into an incubator and exposed to elevated temperatures, steam escapes from the liquid-saturated pores within the lid and creates a saturated atmosphere corresponding to the temperature in the cavity between sample carrier and lid.
  • the porous cover material it is also possible that condensation on the inside of the cover is avoided because condensing Liquid is absorbed sufficiently quickly into the pores and thus a dripping down into individual wells is prevented.
  • a falsification of investigations by dripping liquid is excluded.
  • a lid maintains the required sterility over a long incubation period. This is achieved with a lid with small pore diameters (preferably 0.2-0.5 ⁇ m), which prevents germs from penetrating from the outside. Surprisingly, a sufficient sterility is given even with larger pore diameters.
  • the lid is also designed so that from the outside, no liquid can penetrate inwards.
  • the lid has a shape that can be used very well in automated inspection systems.
  • a porous cover plate has the particular advantage that the atmosphere in the cavity between the sample carrier and the lid by the choice of liquid, with which the porous plate is wetted, can be adjusted.
  • the goal is z. B., that due to a temperature increase resulting vapor pressure change in the cavity between the sample carrier and lid is achieved by evaporation of the liquid contained in the cover plate substantially, thus preventing the evaporation of the samples.
  • This is achieved inter alia by the much larger active surface of the porous cover plate compared to the surface of the sample carrier, so that the evaporation takes place very quickly on the surface of the porous plate and thus the vapor pressure in the cavity is built up in a short time.
  • a liquid is usually selected which has the same or a similar vapor pressure as the sample liquid.
  • an automatic regulation of the vapor pressure takes place in the cavity.
  • a porous film is provided instead of or in addition to the porous cover plate.
  • the porosity of the film also ensures that the saturated atmosphere in the cavity between the receiving part and the lid is substantially maintained under changing environmental conditions.
  • the cover instead of the porous cover plate has a porous film
  • a film can be used as the film, which automatically closes again. This is possible due to the small size of the holes created.
  • films in which the hole is retained since, with a small number of holes, the dimensions of the holes are so small that there is no appreciable impairment of the atmosphere in the cavity between the film and the receiving part.
  • a porous film in addition to a cover plate.
  • the film is preferably arranged between the cover plate and the receiving part. This makes it possible to ensure or adjust the sterility within the sample carrier by a suitable choice of the pore size of the film.
  • the pore size of the cover plate can then be chosen independently of the sterility requirements.
  • a cover plate can then, for example, a textile material, a sponge or other suitable for liquid storage material z. B. in the form of porous flakes or beads are used.
  • the gas permeability of the lid can be controlled by the film used.
  • the pore diameter of the porous cover plate and / or the film are preferably less than 0.5 .mu.m, in particular less than 0.2 .mu.m. As a result, the sterility can be ensured within the sample carrier. Surprisingly enough sterility is achieved even with larger pore diameters.
  • the invention further relates to a lid for a sample carrier, in particular a titer plate.
  • the lid according to the invention preferably has a porous cover plate which is wettable with a liquid for the purpose of regulating the vapor pressure.
  • the cover may be developed according to the preferred embodiments of the sample carrier described above.
  • the cover can serve for receiving electrodes which, for example, extend into individual recesses and cause the migration of sample components in an electric field.
  • the lid has sensors.
  • the moisture content in the cavity between the lid and the receiving part can be determined.
  • a renewed moistening of the lid can be carried out automatically, for example, if the moisture content is too low.
  • the sensors it is possible to arrange the sensors such that the moisture content of the cover plate itself can be determined.
  • sensors for determining substance contents in the cavity With such sensors, for example, the CO 2 content in the cavity can be determined.
  • the lid preferably has handle prisms, so that the lid can be handled automatically by a gripper arm.
  • the handle prisms and the dimensions of the sample carrier preferably correspond to the dimensions and the position of the handle prisms in the case of the sample carrier handled together with the lid. It is thus possible to handle both the lid and the receiving part of the sample carrier with the same gripping arm.
  • the sample carrier is a chip
  • the channels provided in the chip it is preferable for the channels provided in the chip to be micro- or nano-channels whose width and height are in the range from 1 ⁇ m to 1 mm.
  • electrodes can also be arranged within the channels.
  • the sample carrier according to the invention is in particular for the storage of reagents, for the storage of samples, such as cells or other sample substances, for the separation of liquid, for example by means of CE or CEC, for the manipulation of Samples, for example, by electroporation, electropermeation or hybridization and the like. be used.
  • sample carrier or the like a transparent glass plate. so that the samples can be used by fluorescence correlation spectroscopy, especially using convocal optics.
  • sample carriers and an optical tweezers for holding particles can be used.
  • the invention further comprises a method for the preparation of tests of chemical and / or biological micro samples.
  • small samples with a volume of preferably 0.2 to 5 .mu.l in wells of the sample carrier, in particular in wells of the titer plate are introduced.
  • the sample carrier is closed with a lid, which preferably has a porous cover plate.
  • the porous cover plate can be advantageously developed as described above.
  • the porous cover plate is wetted with a liquid. As a result, a vapor pressure regulation is achieved in the cavity formed between the sample top and the lid.
  • the inventive method has the advantages described above with regard to the use of a lid with a porous cover plate.
  • the lid is sterilized prior to sealing the sample carrier. This avoids that germs get into the samples when closing the sample carrier.
  • the liquid used is a PBS buffer containing at least 0.7% of sodium chloride.
  • a buffer solution is used which has substantially the same vapor pressure, like the samples contained in the wells. It is also advantageous to use a liquid having a pH of 7-8, in particular of 7.5.
  • the visible in Fig. 1 cover 10 has a porous cover plate 12 which is supported by a holding part 14.
  • a holding part 14 In the illustrated embodiment, it is a frame-shaped holding part 14 which completely surrounds the cover plate 12.
  • the cover plate 12 may be glued into the holding part 14.
  • supports 16 In order to define the position exactly, supports 16 (FIG. 2) are introduced into the holding part 14.
  • the supports 16 are L-shaped in cross-section and protrude from the frame-shaped holding part 14 inwards, so that the cover plate 12 can be inserted from above into the lid 10 and lies on the supports 16.
  • the pads 16 are preferably also frame-shaped so that they extend completely along the inner edge 18 of the holding part 14. However, individual pads 16 may be arranged distributed on the inner circumference 18 of the frame.
  • a sample carrier 20 (FIG. 2) has a frame-shaped holding part 22 with a rectangular opening. In this a receiving part 24 is arranged with recesses 26. In the wells 26, the samples are housed.
  • the receiving part 24 consists for example of plastic and has recesses 26 through openings. These are on the bottom, preferably with a glass plate 28, closed.
  • the sealing lip 30 is connected via the support 16 with the holding part 14. In the exemplary embodiment shown, this is a preferred embodiment in which the sealing lip extends completely along the inner circumference 32 of the sample carrier 20.
  • the sealing lip 30 is thus rectangular according to the opening in the frame-shaped holding part 22.
  • a cavity 38 is formed between a bottom 34 of the cover plate 12 and a top 36 of the receiving part 24, a cavity 38 is formed.
  • the cavity 38 forms due to very low evaporation of liquid from the wells 26 and due to the main evaporation of the liquid provided in the porous cover plate 12, a vapor pressure.
  • the lower part of the cavity 38 forms a recess in the sample carrier 20. In this recess engages the sealing lip 30 for fixing the position.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Analysing Materials By The Use Of Radiation (AREA)

Claims (17)

  1. Porte-échantillon, en particulier plaque de titrage, comportant
    un élément de réception (24) avec des évidements (26) pour recevoir de très petits échantillons et
    un couvercle (10) fermant l'élément de réception (24) de telle sorte qu'il se forme une cavité (38) entre une surface d'échantillon et le couvercle (10)
    caractérisé en ce que
    le couvercle (10) présente une plaque de recouvrement (12) poreuse, mouillable par un liquide, pour la réception et la distribution de liquide de telle sorte qu'une atmosphère saturée en liquide est maintenue sensiblement dans la cavité (38) dans des conditions ambiantes variables et qu'un échange de gaz est possible à travers la plaque de recouvrement (12) poreuse,
    la cavité (38) étant réalisée entre une face inférieure (34) de la plaque de recouvrement (12) et une face supérieure (36) de l'élément de réception (24).
  2. Porte-échantillon selon la revendication 1, caractérisé en ce que, dans des conditions ambiantes variables, l'atmosphère saturée diverge de moins de 5 %, en particulier de moins de 2 % par rapport à un niveau de saturation maximal.
  3. Porte-échantillon selon la revendication 1 ou 2, caractérisé en ce que le couvercle (10) présente une feuille poreuse.
  4. Porte-échantillon selon la revendication 3, caractérisé en ce que la feuille poreuse est disposée parallèlement à la plaque de recouvrement (12), de préférence entre la plaque de recouvrement (12) et l'élément de réception (24).
  5. Porte-échantillon selon l'une des revendications 1 à 4, caractérisé en ce que la plaque de recouvrement (12) poreuse et/ou la feuille sont supportées par un élément de support (14) entourant en forme de cadre la plaque de recouvrement (12).
  6. Porte-échantillon selon l'une des revendications 1 à 5, caractérisé en ce qu'il est prévu, sur le couvercle (10) et/ou l'élément de support (14) une lèvre d'étanchéité (30) orientée en direction de l'élément de réception (24).
  7. Porte-échantillon selon la revendication 6, caractérisé en ce que la lèvre d'étanchéité (30) est fermée sur elle-même et s'étend de préférence le long du bord (32) de l'élément de réception (24).
  8. Porte-échantillon selon la revendication 6 ou 7, caractérisé en ce que l'élément de réception (24) est entouré par un cadre (22) surélevé de telle sorte qu'il se forme un évidement dans lequel la lèvre d'étanchéité (30) s'engage pour la fixation en position.
  9. Porte-échantillon selon l'une des revendications 1 à 8, caractérisé en ce que les pores de la plaque de recouvrement (12) poreuse et/ou de la feuille présentent un diamètre de pores moyen inférieur à 0,5 µm, en particulier à 0,2 µm.
  10. Porte-échantillon selon l'une des revendications 1 à 9, caractérisé en ce que la plaque de recouvrement (12) est en céramique ou en verre.
  11. Porte-échantillon selon l'une des revendications 1 à 10, caractérisé en ce que le couvercle (10) présente un capteur, en particulier pour déterminer le degré d'humidité et/ou les taux de substance dans la cavité (38).
  12. Couvercle pour un porte-échantillon, en particulier une plaque de titrage, caractérisé par une plaque de recouvrement (12) poreuse qui peut être mouillée avec un liquide pour la régulation de la pression de vapeur.
  13. Procédé de préparation d'analyse de micro-échantillons chimiques et/ou biologiques, comportant les étapes :
    apport des micro-échantillons sur un porte-échantillon, en particulier introduction des micro-échantillons dans les évidements (26) d'une plaque du titrage,
    fermeture du porte-échantillon avec un couvercle (10), le couvercle (10) étant réalisé de telle sorte qu'une atmosphère saturée est sensiblement maintenue dans des conditions ambiantes variables dans une cavité (38) réalisée entre le couvercle et une surface d'échantillon, et
    mouillage du couvercle (10) présentant une plaque de recouvrement (12) poreuse, avec un liquide pour la régulation de l'atmosphère.
  14. Procédé selon la revendication 13, dans lequel le couvercle (10) est stérilisé avant la fermeture du porte-échantillon.
  15. Procédé selon la revendication 13 ou 14, caractérisé en ce que le liquide utilisé est un tampon PBS contenant au moins 0,7 % de chlorure de sodium.
  16. Procédé selon l'une des revendications 13 à 15, caractérisé en ce que le liquide utilisé présente une valeur de pH de 7-8, en particulier de 7,5.
  17. Procédé selon l'une des revendications 13 à 16, dans lequel avant ou après l'introduction de l'échantillon, on introduit une solution réactive ou une solution nutritive dans les évidements (26).
EP01997350A 2000-11-23 2001-11-23 Porte-echantillon, son couvercle et procede de preparation d'analyse Expired - Lifetime EP1341611B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE10058108A DE10058108A1 (de) 2000-11-23 2000-11-23 Probenträger, Deckel für Probenträger und Verfahren zur Untersuchungsvorbereitung
DE10058108 2000-11-23
PCT/EP2001/013651 WO2002041993A1 (fr) 2000-11-23 2001-11-23 Porte-echantillon, son couvercle et procede de preparation d'analyse

Publications (2)

Publication Number Publication Date
EP1341611A1 EP1341611A1 (fr) 2003-09-10
EP1341611B1 true EP1341611B1 (fr) 2007-09-26

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EP01997350A Expired - Lifetime EP1341611B1 (fr) 2000-11-23 2001-11-23 Porte-echantillon, son couvercle et procede de preparation d'analyse

Country Status (4)

Country Link
EP (1) EP1341611B1 (fr)
AT (1) ATE374077T1 (fr)
DE (2) DE10058108A1 (fr)
WO (1) WO2002041993A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8222048B2 (en) 2007-11-05 2012-07-17 Abbott Laboratories Automated analyzer for clinical laboratory

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DE10352716A1 (de) * 2003-11-05 2005-06-16 Einsle, Xaver Plattform
DE102004041941B4 (de) * 2004-08-30 2007-01-11 P.A.L.M. Microlaser Technologies Ag Verfahren zur Gewinnung von biologischen Objekten mit einer Aufnahmeeinheit
DE102006048264B4 (de) * 2006-10-12 2011-03-24 Bundesrepublik Deutschland, vertreten durch das Bundesministerium der Verteidigung, vertreten durch das Bundesamt für Wehrtechnik und Beschaffung Verfahren zum Nachweis spezifischer Nukleinsäuresequenzen
DE102011004449B4 (de) * 2011-02-21 2019-01-24 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Biopsieplättchen, Einbettkassette und Diagnosevorrichtung

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EP0388159A3 (fr) * 1989-03-15 1991-06-12 Seiko Instruments Inc. Dispositif pour le scellement d'un liquide dans des cavités
US5342581A (en) * 1993-04-19 1994-08-30 Sanadi Ashok R Apparatus for preventing cross-contamination of multi-well test plates
WO1995027196A1 (fr) * 1994-04-04 1995-10-12 Sanadi Ashok R Procede et appareil destine a empecher la contamination croisee de plaques de test a cupules multiples
US5789251A (en) * 1994-06-16 1998-08-04 Astle; Thomas W. Multi-well bioassay tray with evaporation protection and method of use
BE1010984A3 (nl) * 1995-02-17 1999-03-02 Praet Peter Van Incubator voor microtiter plaat.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8222048B2 (en) 2007-11-05 2012-07-17 Abbott Laboratories Automated analyzer for clinical laboratory
US9329194B2 (en) 2007-11-05 2016-05-03 Abbott Laboratories Automated analyzer for clinical laboratory

Also Published As

Publication number Publication date
DE50113066D1 (de) 2007-11-08
WO2002041993A1 (fr) 2002-05-30
EP1341611A1 (fr) 2003-09-10
DE10058108A1 (de) 2002-06-06
ATE374077T1 (de) 2007-10-15

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