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EP2129467B1 - Pulvérisateur et procédé de préparation du traitement d'un échantillon biologique - Google Patents

Pulvérisateur et procédé de préparation du traitement d'un échantillon biologique Download PDF

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Publication number
EP2129467B1
EP2129467B1 EP08735665.5A EP08735665A EP2129467B1 EP 2129467 B1 EP2129467 B1 EP 2129467B1 EP 08735665 A EP08735665 A EP 08735665A EP 2129467 B1 EP2129467 B1 EP 2129467B1
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EP
European Patent Office
Prior art keywords
sample
vessel
refrigerated
process according
movable body
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.)
Not-in-force
Application number
EP08735665.5A
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German (de)
English (en)
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EP2129467A2 (fr
Inventor
Dennis Mertens
Thomas Voit
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.)
Qiagen GmbH
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Qiagen GmbH
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Filing date
Publication date
Application filed by Qiagen GmbH filed Critical Qiagen GmbH
Publication of EP2129467A2 publication Critical patent/EP2129467A2/fr
Application granted granted Critical
Publication of EP2129467B1 publication Critical patent/EP2129467B1/fr
Not-in-force legal-status Critical Current
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B02—CRUSHING, PULVERISING, OR DISINTEGRATING; PREPARATORY TREATMENT OF GRAIN FOR MILLING
    • B02C—CRUSHING, PULVERISING, OR DISINTEGRATING IN GENERAL; MILLING GRAIN
    • B02C17/00—Disintegrating by tumbling mills, i.e. mills having a container charged with the material to be disintegrated with or without special disintegrating members such as pebbles or balls
    • B02C17/14—Mills in which the charge to be ground is turned over by movements of the container other than by rotating, e.g. by swinging, vibrating, tilting

Definitions

  • the invention relates to a method for the preparation of a vegetable or animal sample for a processing, that is, for example, for the isolation of nucleic acids or proteins from the sample and a pulverizer.
  • a processing that is, for example, for the isolation of nucleic acids or proteins from the sample and a pulverizer.
  • Such preparations and examinations are carried out in a laboratory by a laboratory assistant or a laboratory technician on the basis of standardized work instructions.
  • Such a manual includes a so-called protocol.
  • An example of such a protocol for the isolation of plasmid DNA from E. coli. goes out of the document DE 101 53 957 A1 out.
  • kits can be obtained commercially, for example the "UItraClean Tissue DNA Isolation Kit” of the company, depending on the sample and the desired result.
  • Qiagen www.Qiagen.com
  • a sample is processed using such a kit according to a given protocol, it must be suitably prepared.
  • an organ is taken from a test animal, such as a rat. It depends on the objective, which organ of an animal is selected.
  • the removed tissue of the animal is washed in a washing buffer solution, for example in PBS (Phosphate Buffered Saline with the following contents: Na 2 HPO 4 (dried), NaH 2 PO 4 (dried), NaCl and distilled water).
  • a washing buffer solution for example in PBS (Phosphate Buffered Saline with the following contents: Na 2 HPO 4 (dried), NaH 2 PO 4 (dried), NaCl and distilled water).
  • tissue with a body temperature of, for example, 37 ° C is immersed in liquid nitrogen. Bubbles develop. The tissue is only removed from the liquid nitrogen when bubbling stops. Subsequently, the tissue is stored at -80 ° C, for example, with the aid of dry ice.
  • RNAlater® is a viscous liquid developed by Ambion (www.ambion.com) for the preservation of fresh tissue. The preservative effect is based primarily on the fact that all enzymes inactivated by dehydration in the tissue and cell activities are stopped. The viscous fluid must quickly diffuse into all cells of the tissue. The size of the fabric pieces is therefore limited to an edge length of a maximum of half a centimeter. Following the chemical treatment, the tissue thus treated is also cooled at -80 ° to store it until processing.
  • the separated sample ie the separated tissue is now digested, that is, the cell walls must be opened.
  • This can be done mechanically, chemically or enzymatically.
  • Mechanical disruption takes place, for example, with the aid of a "TissueRuptor” from Qiagen, which is known from the TissueRuptor Handbook, July 2006 by Qiagen.
  • a rotating knife with 35,000 revolutions per minute smashes cell walls of the tissue.
  • Mechanical digestions are regularly performed in a buffer to avoid damage to ingredients such as nucleic acids.
  • a sample is first blocked in paraffin and then cut into thin tissue layers by means of a microtome.
  • scalping can only be done with soft materials such as leaves, soft beans, etc.
  • soft materials such as leaves, soft beans, etc.
  • they are cooled in liquid nitrogen and ground in a liquid nitrogen cooled mortar using a pestle.
  • German patent 738 286 teaches to freeze and grind cells together with a dispersion liquid to thus crush cells.
  • a crushing / mixing device for foods such as spices.
  • the device comprises a hollow body with a ball therein, with which foodstuffs are to be crushed.
  • JP 2002-066366 A such as JP 03-186 360 A disclose further examples of ball mills and comparable devices.
  • a solution to the problem comprises the features of claim 1. Further solutions include the features of the dependent claims, which are also directed to a method. An apparatus for carrying out the method comprises the features of the last subsidiary claim. Advantageous embodiments emerge from the subclaims.
  • the claimed teaching works for both plant and animal or human tissue, both for stabilized and fresh samples of plants or tissues.
  • a sample is first washed as described above and treated, for example, with liquid nitrogen or stored in RNAlater.
  • a closable preferably made of metal vessel or plastic existing vessel, which is equipped with a metal inlay, and a container located in the body, movable body cooled to a temperature which is well below 0 ° C.
  • the plastic container is a shell for the inlay.
  • An inlay is a body made separately from the shell.
  • a double-walled vessel as for example from the document US 6,235,501 B1 10, which does not comprise an inlay in the sense of the present invention, since the two vessel walls known therefrom are integrally connected to one another.
  • the temperature to which the vessel is cooled should be at least -50 ° C.
  • a temperature of about - 80 ° C for example, from -70 ° C to -90 ° C to choose, since this is much further away from 0 ° C.
  • -80 ° C or -70 ° C to -90 ° C can be provided inexpensively by means of dry ice. It has been shown that can be achieved by cooling to about -80 ° C particularly good results. Temperatures lower than -80 ° C are possible to a certain extent. It should be noted, however, that the vessel must not be allowed to cool too much. For example, a temperature of liquid nitrogen, ie -196 ° C, has proven to be too low for good results.
  • the interior of the vessel is dimensioned and designed in addition to the movable body therein.
  • Particularly suitable is a cylindrical interior with hollow spherical ends.
  • the movable body is then preferably a ball or a bolt with spherical ends.
  • the diameter the ball or the bolt is smaller than the diameter of the interior, so as to ensure the mobility.
  • the movable body is made of a hard, preferably heavy material such as metal in order to be able to crush the sample.
  • the biological sample for example a cooled rat heart, is preferably placed completely in the vessel without first chopping the sample in order to obtain good results.
  • a sample at least -50 ° C cold is added to the vessel.
  • the vessel is then shaken, in particular by hand, in particular for 10 to 30 seconds so that the movable body is hurled back and forth.
  • the thus crushed sample is removed and the desired amount is processed using a kit according to a protocol.
  • the sample is in powder form, it advantageously has a particularly large surface area, from which subsequently used chemicals can attack.
  • a desired amount of powder can be provided particularly easily for subsequent steps, for example by weighing or even by appropriately sized measuring vessels, such as a measuring spoon.
  • FIG. 1 shows in section a vessel 1, on which a cover 2 can be screwed.
  • a ball 3 which has a smaller diameter compared to the diameter of the vessel 1.
  • the diameter of the vessel is in the range of a few centimeters.
  • the interior of the vessel 1 is cylindrical. When closed, the ends are 5 and 6 of the vessel hemispherical shaped. By appropriate shaking of the vessel 1, the ball 3 is moved back and forth in the interior and strikes the ends 5 and 6.
  • the vessel 1, the lid 2 and the ball 3 are made of metal and that made of stainless steel.
  • the vessel is cooled in dry ice at -80 ° C.
  • a sample 7 is taken from an animal.
  • the sample is washed and dipped in nitrogen until no more bubbles are formed or stored in RNAlater for 24 hours, for example. Following this, the sample can first be stored in dry ice at -80 ° C. In the case of a plant dried plant tissue such as seeds in the vessel 1 is cooled on dry ice. Fresh plant material such as leaves are previously cooled to -80 ° C.
  • the vessel If the vessel has been cooled to the desired temperature, it is removed by hand.
  • the hand is expediently protected from the cold with a cotton glove and a glove made of latex.
  • the sample 7 With a pair of tweezers, the sample 7 is placed in the vessel 1, in which the ball 3 is located. Subsequently, the lid 2 is screwed onto the vessel 1 and this closed so tightly. Now the vessel 1 is shaken by hand back and forth, so that the ball 3 impinges alternately on the two ends 5 and 6. The sample 7 is thereby smashed. This process ends after 20 to 30 seconds.
  • the result is a free-flowing powder that can be initially stored in the vessel 1 further. It is unscrewed immediately or following storage of the lid 2 and removed the crushed sample. A desired amount can now be provided by weighing, for example.
  • liver was taken from a rat and washed in PBS. Following this, the liver was stored in RNAlater® for 24 hours. Following this, the rat liver was stored at -80 ° C until further use.
  • the liver was crushed in the cooled, closed vessel for 20 seconds as previously described. With the aid of a funnel, the crushed sample was filled into a Falcon tube and stored in dry ice at -80 ° C initially.
  • the used DNeasy protocol from Qiagen includes the following steps.
  • the 1 0 mg tissue is incubated with 180 ⁇ l of BUFFER 1 + 20 ⁇ l proteinase K at 56 ° C for one hour in a rocking platform.
  • 4 ⁇ l RNase A are pipetted in and incubated for 2 minutes at room temperature.
  • 200 ⁇ l of AL lysis buffer
  • 200 ⁇ l of ethanol 100%) are pipetted in and mixed by gentle inversion.
  • the point where the DNA can be recognized as a "cloud”.
  • the resulting solution is mixed with the pipette 3 times by drawing.
  • the entire solution is then pipetted into a DNeasy column and centrifuged for 1 minute at 8000 rpm.
  • the corresponding nucleic acid is now bound to the column material.
  • the supernatant is discarded and the column now with 500 ul wash buffer AW 1 at 8000 revolutions Washed for 1 minute per minute. This step was additionally applied again, even if it is not recommended by the protocol. Subsequently, the column is washed as described in the protocol with 500 ul buffer AW 2 for 3 minutes at 14,000 revolutions per minute.
  • the supernatant is again discarded.
  • the optional drying step described in the protocol is used for 1 minute at 14,000 rpm.
  • 200 ⁇ l of RNase-free water is pipetted into the center of the column and incubated at room temperature for 1 minute, then centrifuged at 8000 rpm for 1 minute. This eluate is measured by means of a spectrometer. This serves to determine the concentration and to determine the degree of purity.
  • an agarose gel is made for qualitative and quantitative DNA determination. On this basis, the quality of the DNA, the degree of degradation, the size of the molecule and its amount can be determined.
  • the bright upper areas in the FIG. 2 characterize the genomic DNA.
  • the bright area above TR is not as high as above TD +, TD- or TD / TR.
  • the Fig. 2 shows images of DNA agarose gels used as described above to determine the quantity and quality of the DNA.
  • a 1% gel is poured.
  • 1 g of agarose powder (long-chain carbohydrate molecules that can polymerize) is weighed and dissolved in 100 ml (1 x) TAE buffer and heated in the microwave.
  • the resulting solution is mixed with 5 .mu.l Ethid iumbromid and mixed by shaking the flask and poured into a mold.
  • the hot solution now combs are clamped, which leave as the solution cools, which polymerizes, prints in the gel, which later form the slots in which the sample mixed with 5 ⁇ l Loading Dye is pipetted into it.
  • a marker is additionally added to the gel for later size comparison.
  • the DNA is distributed after application of an electrical voltage due to their different weight and charge, ie small degraded fragments move faster in the gel than high molecular weight DNA.
  • the DNA obtained by the method according to the invention can be seen in the form of a cloud with the naked eye.
  • the same method was used for pulverization as in the experiment described above.
  • the rat's heart was removed and cooled by liquid nitrogen until no more bubbles were produced. Afterwards the heart was stored on dry ice (-80 ° C). Shortly before processing, the jar 1 was cooled on dry ice and filled with the frozen whole heart and shaken by hand for 20 seconds. The powder was then stored in the jar to allow time to get the materials for the RNeasy Fibrous Tissue Mini Protocol. This poses no problem, because the vessel was digestion and storage vessel in one.
  • the supernatant was then pipetted into a new 2 ml reaction tube and 0.5X volume of 96-100% ethanol pipetted in and mixed through the pipette. Subsequently, first 700 ⁇ l of the solution were pipetted into the RNeasy column and centrifuged for 15 seconds at 10,000 revolutions per minute. Since the column has only 700 ⁇ l capacity, this step was repeated for the remaining solution so that the entire sample was passed over the column. The supernatant after centrifugation was discarded. Thereafter, the column was washed with 350 ⁇ l of buffer RW1 for 15 seconds at 8000 g and the supernatant discarded.
  • the column was transferred to a new 2 ml reaction vessel and filled with 30 ⁇ l of RNase-free water and centrifuged for 1 minute at 10,000 revolutions per minute to elute. The eluate was then also measured by spectrometer and evaluated by RNA agarose gel.
  • FIG. 3 shows a particularly preferred embodiment of a closable vessel with which a sample is pulverized.
  • This vessel is called pulverizer below. It comprises an inlay 10, which preferably consists of metal for the reasons stated above.
  • the inlay 10 is comprised of a shell 11, preferably made of plastic.
  • the plastic shell serves primarily as an insulator to maintain the low temperature during shaking. Overall, the weight can be advantageously reduced in comparison to a completely made of metal pulverizer, which facilitates handling.
  • the double arrow below the in FIG. 3 The pulverizer shown illustrates the preferred direction of movement to comminute the sample 7.
  • the buffers and reagents mentioned in the present application can be obtained commercially from Qiagen GmbH, Hilden, Germany, unless otherwise specified.
  • FIG. 4 shows a section through a further improved embodiment of the invention.
  • the pulverizer comprises an inner sealable container 4 made of plastic.
  • the plastic inner container adjacent to so closely to the preferably made of metal inner walls of the pulverizer, so that the existing plastic inner container can not be destroyed by a ball 3 or a similar means during pulverization.
  • the plastic inner container 4 is removed after pulverization and can now serve as a storage vessel. Since the inner container is made of plastic, this can be made very inexpensive and therefore suitable for single use.
  • the lid of the inner container closes the remaining part of the container preferably by a positive connection, or it can be screwed onto the remaining part.
  • a positive connection is possible because plastic can be sufficiently elastic, so that, for example, an inwardly projecting bead of the lid snaps into a designated annular recess of the rest of the container body or vice versa, when the lid is pressed to the rest of the container body suitable.
  • Such a positive connection is particularly preferable because accidental release of the lid is particularly reliably avoided.
  • a container can be closed faster compared to a closure with a screw cap.
  • a set comprises, in addition to a pulverizer, a multiplicity of inner containers, which are preferably optically different, for example because of differently colored covers or different embossments.
  • the different optics can be used advantageously for the identification of a content.
  • a red-colored container lid can be used to identify a "heart" located therein and another, for example, green-colored lid another organ, such as a lung.
  • An internal, the storage of a powdered sample serving container must then no longer be labeled separately, which can be problematic due to the low temperatures.
  • the existing plastic plastic inner container can be made of PET, but also made of PP or PE, since such plastics are grown to the intended low temperatures in principle.
  • a kit in one embodiment, includes one or more measuring spoons in addition to a pulverizer so as to facilitate or accelerate the removal of the required amount of sample from the pulverizer or from an inner container.
  • a measuring spoon is so dimensioned and assigned to certain samples that the measuring spoon is capable of receiving a suitable amount of sample for further processing. A separate weighing of a sample taken from the pulverizer can thus be accelerated or even completely eliminated.
  • a set comprises a plurality of optically labeled measuring spoons associated with different samples.
  • a red-colored measuring spoon can be provided for the removal of a heart sample.
  • the identifications of inner containers or parts of inner containers coincide with the identifications of measuring spoons.
  • Such a set then also includes a predetermined assignment of identifiers to samples, that is, for example, to organs. If, for example, a green-colored measuring spoon is provided for the organ lung, then an inner container is likewise completely or partially dyed green. The dimensioning of the measuring spoon is adapted to the sample "lung".
  • the set then contains a corresponding assignment rule. This may consist in that already a corresponding sample, so for example a lung on the corresponding container and / or the measuring spoon is shown.
  • a set comprises, in addition to a pulverizer, a shaking apparatus to facilitate the opening of an imprinter Automated perform sample located pulverizer automatically.
  • a shaking apparatus to facilitate the opening of an imprinter Automated perform sample located pulverizer automatically.
  • One or more Pulverisatoren can be used in the shaker or attached to this suitable.
  • the shaking apparatus comprises coolable receiving devices for receiving pulverizers.
  • the shaking apparatus does not need to be digested immediately after filling the sample in the pulverizer, if it can be sufficiently cooled.
  • a large number of pulverizers can be prepared and used before a large number of samples are then automatically digested at the same time.

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  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (25)

  1. Procédé de digestion d'un échantillon biologique par l'introduction d'un échantillon biologique solide ayant une température inférieure à -50°C dans un récipient réfrigéré au-dessous de -50 °C qui peut être fermé, dans lequel se trouve un corps mobile réfrigéré au-dessous de -50 °C qui est mis en mouvement après que le récipient est fermé et qui triture l'échantillon biologique, caractérisé par le fait que le récipient n'est pas réfrigéré pendant la trituration et qu'aucun milieu de refroidissement ou aucune substance chimique est introduit dans le récipient pendant ou avant la digestion.
  2. Procédé selon la revendication 1, caractérisé par le fait que le récipient est une douille qui peut être ouvert et fermé, et un inlay.
  3. Procédé selon la revendication 1 ou 2, caractérisé par le fait que l'échantillon biologique ayant une température dans la plage de -70 °C à -90 °C est introduit dans un récipient réfrigéré à une température dans la plage de -70 °C à -90 °C qui peut être fermé.
  4. Procédé selon l'une des revendications 1 à 3, caractérisé par le fait que l'échantillon biologique est un échantillon végétal.
  5. Procédé selon l'une des revendications 1 à 4, caractérisé par le fait que l'échantillon biologique est un tissu d'origine animal ou humain.
  6. Procédé selon l'une des revendications 1 à 5, caractérisé par le fait que le corps mobile est fait bouger par agitation manuelle.
  7. Procédé d'analyse d'un échantillon biologique comprenant les étapes consister à digestion l'échantillon selon l'une des revendications 1 à 6, enlever l'échantillon ouvragé partiellement ou complètement et isoler des acides nucléiques et/ou des protéines de l'échantillon ouvragé.
  8. Procédé de préparation du traitement d'un échantillon d'origine animal ou humain, dans lequel l'échantillon est tout d'Abord lavé et ensuite est stabilisé, dans lequel l'échantillon suite à la stabilisation est surgelé, et est préférablement surgelé à -50 à 110 °C, dans lequel un récipient qui peut être fermé (1, 2) ayant un corps mobile (3) là-dedans est surgelé, et est préférablement surgelé à -50 à 110 °C, dans lequel l'échantillon surgelé est mis dans le récipient surgelé ayant un corps mobile (3) là-dedans, par la suite le récipient surgelé est fermé et le récipient surgelé est agité de gauche à droite et à la suite une échantillon trituré de ce fait est enlevé, le récipient n'étant pas surgelé pendant la trituration et aucun milieu de refroidissement ou substance chimique étant introduit dans le récipient pendant ou avant l'digestion.
  9. Procédé selon l'une des revendications précédentes, dans lequel l'échantillon suite à la stabilisation est surgelé dans la glace sèche et/ou le récipient qui peut être fermé (1, 2) ayant un corps mobile (3) là-dedans est surgelé dans la glace sèche, et est préférablement surgelé à -80 °C.
  10. Procédé selon l'une des revendications précédentes, dans lequel le récipient surgelé qui peut être fermé ayant l'échantillon (7) et le corps mobile (3) là-dedans est agité de gauche à droite pendant 10 à 40 secondes.
  11. Procédé selon l'une des trois revendications précédentes, dans lequel le récipient ne contient pas des constituants liquides, tel que l'azote liquide ou une solution de tampon, e particulièrement ne contient pas des constituants liquides pendant que l'échantillon (7) est dans le récipient (1, 2) et/ou l'échantillon n'est pas les os ni la peau.
  12. Procédé selon l'une des quatre revendications précédentes, dans lequel un foie, une cour, des feuilles ou des graines végétales sont utilisés en tant qu'échantillon.
  13. Procédé selon l'une des cinq revendications précédentes, dans lequel le corps mobile (3) est une bille et l'intérieure du récipient qui peut être fermé est en forme cylindrique ayant des bouts (5, 6) en forme de billes creuses.
  14. Procédé selon l'une des six revendications précédentes, dans lequel suite à l'enlèvement de l'échantillon trituré surgelé du récipient qui peut être fermé l'échantillon trituré (7) est traité.
  15. Procédé selon l'une des revendications précédentes, dans lequel l'échantillon ensemble avec le corps mobile (3) est mis dans un conteneur intérieur consistant en matière plastique et le conteneur intérieur (4) est inséré dans le récipient qui peut être fermé-
  16. Pulvérisateur pour des échantillons biologiques pour la mise en oeuvre d'un procédé selon l'une des revendications précédentes comprenant un récipient formé en tant qu'une douille, qui peut être ouvert et fermé, un inlay (10) et un corps mobile (3).
  17. Pulvérisateur selon la revendication 16 caractérisé par le fait que la douille (11) est fabriqué de matière plastique.
  18. Pulvérisateur selon les revendications 16 ou 17, caractérisé par le fait que l'inlay (10) est fabriqué de métal.
  19. Pulvérisateur selon l'une des revendications 16 à 18, caractérisé par le fait que le corps mobile (3) est fabriqué de métal ou d'un minéral.
  20. Pulvérisateur selon l'une des revendications 16 à 19, caractérisé par le fait que le corps mobile (3) est sphérique et le pulvérisateur a un intérieur ovale.
  21. Pulvérisateur selon l'une des revendications précédentes 16 à 20 ayant un récipient (4) consistant en matière plastique.
  22. Kit comprenant un pulvérisateur selon l'une des revendications 16 à 21 ayant une pluralité de conteneurs (4) intérieurs consistant en matière plastique et/ou d'une pluralité de cuillères graduées.
  23. Kit selon l'une des revendications précédentes, dans lequel les conteneurs intérieurs (4) et/ou les cuillères graduées sont optiquement marqués en façon différente.
  24. Kit selon l'une quelconque des deux revendications précédentes ayant une assignation prédéterminée entre des marquages optiques de conteneurs intérieurs et/ou de cuillères graduées et d'échantillons.
  25. Kit selon l'une des revendications précédentes 20 à 23 comprenant un appareille vibreur pour la digestion d'un échantillon qui est dans le pulvérisateur.
EP08735665.5A 2007-04-04 2008-04-02 Pulvérisateur et procédé de préparation du traitement d'un échantillon biologique Not-in-force EP2129467B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102007016221A DE102007016221A1 (de) 2007-04-04 2007-04-04 Pulverisator und dazugehöriges Verfahren für die Vorbereitung zur Prozessierung einer biologischen Probe
PCT/EP2008/053896 WO2008122550A2 (fr) 2007-04-04 2008-04-02 Pulvérisateur et procédé de préparation du traitement d'un échantillon biologique

Publications (2)

Publication Number Publication Date
EP2129467A2 EP2129467A2 (fr) 2009-12-09
EP2129467B1 true EP2129467B1 (fr) 2014-07-02

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EP08735665.5A Not-in-force EP2129467B1 (fr) 2007-04-04 2008-04-02 Pulvérisateur et procédé de préparation du traitement d'un échantillon biologique

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US (2) US8348183B2 (fr)
EP (1) EP2129467B1 (fr)
JP (1) JP5150880B2 (fr)
DE (1) DE102007016221A1 (fr)
WO (1) WO2008122550A2 (fr)

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WO2020176644A2 (fr) * 2019-02-26 2020-09-03 SPEX SamplePrep, LLC Homogénéisateur et procédé de broyage de grandes quantités d'échantillon
JP7416496B1 (ja) * 2023-03-03 2024-01-17 株式会社小泉製作所 粉砕器具
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US20130026268A1 (en) 2013-01-31
US8348183B2 (en) 2013-01-08
EP2129467A2 (fr) 2009-12-09
WO2008122550A3 (fr) 2009-03-19
US20100137567A1 (en) 2010-06-03
JP5150880B2 (ja) 2013-02-27
DE102007016221A1 (de) 2008-10-09
WO2008122550A2 (fr) 2008-10-16
JP2010523956A (ja) 2010-07-15

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