Retention device for a reaction vessel unit

Abstract

A retention device for a reaction vessel unit for centrifuging reagents in the reaction vessel unit, which has at least one blind-shaped reaction vessel with a reaction vessel opening, wherein the retention device is designed to be placed on or inserted into the at least one reaction vessel and either has one or more through-openings that are smaller than the reaction vessel opening, so that when centrifuging with the reaction vessel opening facing radially outwards, a solid body that is larger than the through-openings can be retained by the retention device, or that at least one solid body for immobilizing one or more chemical or biological samples or which itself constitutes a sample is attached to the retention device in such a way that the solid body is located inside the at least one reaction vessel when the retention device is placed on a reaction vessel unit.

Description

[0001] The present invention relates to a retaining device for a reaction vessel unit.

[0002] It is known that reaction vessel units containing multiple reaction vessels, such as microtiter plates (MTPs), along with their reaction vessels or wells, can be cleaned by centrifugation. The microtiter plates are mounted on a rotor in a centrifuge's rotor chamber with the openings of the reaction vessels facing away from the rotor's rotational axis, and are centrifuged at a speed of up to several thousand rpm. This causes the contents of the reaction vessels to be ejected and collected by a wall of a rotor chamber containing the rotor.

[0003] WO 2023 / 148272 A1 and WO 2023 / 148273 A1 each disclose collecting devices for a reaction vessel unit. These devices have a collecting surface configured such that, in a centrifuge, it is arranged opposite the openings of the reaction vessels, so that during centrifugation, the ejected liquid is drained away in a controlled manner along the collecting surface approximately parallel to the rotor's rotational axis. With such a device, the ejected liquid can be drained from the rotor chamber in a controlled manner without the liquid being distributed throughout the rotor chamber and posing a risk of contamination for other reaction vessel units to be centrifuged. This can significantly reduce the need for cleaning the rotor chamber.

[0004] DE 20 2014 010 544 U1 and DE 10 2021 124 023 A1 disclose centrifuges with elements for cleaning the rotor chamber. These are no longer necessary with such a collecting device, at least not to this extent.

[0005] DE 10 2008 242 971 A1 discloses a centrifuge in which a magnetic device is integrated in order to hold magnetizable particles within a reaction vessel by means of the magnetic force, while liquid is removed from the magnetizable particles during centrifugation.

[0006] WO 2023 / 110944 A1 discloses a reaction vessel unit with reaction vessels designed to retain at least part of the contents during centrifugation due to adhesion forces between the liquid and a retention area and cohesion within the liquid, thus removing only a portion of the liquid from the reaction vessel unit. This retention area can be formed by a special surface coating or by a capillary cavity.

[0007] The present invention is based on the object of providing a retaining device for a reaction vessel unit for centrifuging reagents.

[0008] The object is achieved by a restraint device having the feature of claim 1. Advantageous embodiments are specified in the subclaims.

[0009] The invention relates to a retaining device for a reaction vessel unit for centrifuging reagents in the reaction vessel unit, which has at least one blind-hole-shaped reaction vessel with a reaction vessel opening, wherein the retaining device is designed to be placed on or inserted into the at least one reaction vessel and has either one or more through-openings that are smaller than the reaction vessel opening, so that during centrifugation with the reaction vessel opening directed radially outwards, a solid body that is larger than the through-openings can be retained by the retaining device, or at least one solid body for immobilizing one or more chemical or biological samples or which itself represents a sample is attached to the retaining device in such a way that the solid body is located within the at least one reaction vessel when the retaining device is placed on a reaction vessel unit.

[0010] The invention makes it possible for the first time to arrange any solid in a reaction vessel of a reaction vessel unit and to centrifuge it with a centrifuge in such a way that the reaction vessel opening points away from the axis of rotation, so that any liquid contained in the reaction vessel is ejected, while the solid is retained. This allows solids to be automatically separated from liquids. These solids are usually solids to which a sample reagent is chemically or physically immobilized or coupled. The retention device then allows the sample reagent to be automatically brought into contact with different liquids in succession by dispensing liquid into the reaction vessel and centrifuging, and then separated from them again.This allows any chemical and / or biological test series that are carried out with a sample reagent coupled to a solid to be automated.

[0011] The at least one solid body can also be attached directly to the retaining device, so that the solid body is located within the at least one reaction vessel when the retaining device is placed on a reaction vessel unit.

[0012] The retaining device therefore preferably has the form of an approximately flat plate on which one or more approximately tubular projections are formed, each of which can extend into a reaction vessel of the reaction vessel unit, wherein a grid with the one or more through openings is formed at the end of the tubular projections remote from the flat plate in order to retain the solid body, or the solid body itself is directly attached thereto.

[0013] The solid body(s) can be designed to immobilize one or more chemical or biological samples, or they can themselves represent a sample. Solid bodies for immobilizing chemical or biological samples are known. These can be formed, for example, as spherical plastic bodies called beads or from tissue pieces. Predefined functional groups, such as COOH or NH2, are often coupled to the surface of the solid body, allowing chemical and / or biological samples to be chemically bound therein.

[0014] The solids themselves can serve as samples. Fabrics can be subjected to specific chemical tests. For example, specifically soiled fabrics can be used to test detergents and test corresponding cleaning processes on a small scale.

[0015] The projections of the retaining device, which each dip into one of the reaction vessels during use, are preferably arranged in the same grid as the corresponding reaction vessels of the reaction vessel unit. If the reaction vessel units are microtiter plates, the arrangement of the reaction vessels and thus their grid is standardized. Microtiter plates are available with, for example, 12, 24, 48, 96, 192, 384, or 768 reaction vessels. Accordingly, there are corresponding grids for the arrangement of the reaction vessels, since the base area of ​​the microtiter plates is always the same size at 85.48 mm x 127.76 mm.

[0016] In research and development, there is a significant need to carry out chemical and / or biological reactions using a fully automated workflow. The higher the level of automation, the more samples can be screened in parallel.

[0017] There are a variety of different tests and experiments that are carried out with a sample reagent coupled to a solid.

[0018] For the purposes of the present invention, any body that can be retained by such a retention device during centrifugation is a solid. Liquids and gases are not solids. A solid within the meaning of the present invention does not necessarily have to be rigid. Suitable solids include, for example, pieces of textile or paper. However, any other solids can also be used, such as plastic beads.

[0019] The retaining device can be designed to cover multiple reaction vessels of the reaction vessel unit. The reaction vessel openings are preferably arranged in a common opening plane, so that the retaining device then forms a "perforated" lid for the reaction vessel unit. The retaining device is preferably designed in sections to complement the shape of a corresponding reaction vessel unit, so that it can be positioned in a specific position on the reaction vessel unit. For this purpose, corresponding projections and recesses can be provided, for example, on the retaining device and / or on the reaction vessel unit, which interact to position the reaction vessel unit and the retaining device.

[0020] The retaining device can also be designed as an insert element for insertion into the at least one reaction vessel of the reaction vessel unit. The retaining device can therefore be an insert that is completely received in the reaction vessel. The retaining device can be designed to be flat, wherein the reaction vessel has one or more projections along a trajectory circumferentially extending on the reaction vessel unit, such that the retaining device can be pressed into the reaction vessel up to this projection(s) and is fixed there by friction. However, the retaining device can also be designed from an approximately flat section, on the edge of which one or more fixing elements are arranged, extending approximately transversely to the plane of the flat section and causing a frictional engagement with the inner surface of the respective reaction vessel in order to fix the retaining device in the reaction vessel.

[0021] The retaining device can be configured with a complementary shape to at least one section of the at least one reaction vessel. Preferably, the retaining device has a plurality of such complementarily shaped sections that fit into a plurality of different sections of different reaction vessels of the reaction vessel unit. These complementarily shaped sections of the retaining device can, on the one hand, establish a frictional connection with the reaction vessel unit to fix the retaining device to the reaction vessel unit, and / or, on the other hand, ensure a precise positioning of the retaining device on the reaction vessel unit.

[0022] The restraint device can be designed as a grid with wire-shaped grid struts. "Wire-shaped" refers to elongated, thin struts. These can have a circular cross-section. However, they can also have a polygonal cross-section. These grid struts are preferably arranged crosswise, and in a plan view of the grid, they can be arranged at different angles to each other. A grid in which the wire-shaped grid struts are arranged at right angles to each other is expedient.

[0023] The grid struts preferably have a maximum thickness of 1 mm, in particular a maximum thickness of 0.5 mm and particularly preferably a maximum thickness of 0.3 mm.

[0024] The open area of ​​the passage opening is preferably not more than 15 mm 2 or 10 mm 2 or 3 mm 2 or 2 mm 2 or 1 mm 2 or 0.5 mm2 The smaller the through-hole, the smaller the solid body that can be retained by the retaining device.

[0025] The area of ​​the through hole is preferably at least 0.5 mm 2 or 1 mm 2 or 2 mm 2 or 5 mm 2 or 10 mm 2 or 15 mm 2 The larger the opening of the through-hole, the easier it is to direct a jet of liquid into the reaction vessel through the opening during dispensing.

[0026] The reaction vessels have a bottom and circumferential side walls. The surfaces of the retaining device, which are arranged facing the bottom, are preferably predominantly inclined to a horizontal extent of the reaction vessel unit. The reaction vessel units generally comprise several reaction vessels arranged side by side in a plane. This plane defines the horizontal extent. These inclined surfaces can, for example, be formed by lattice struts with a circular cross-section, which have only an infinitesimally small section running parallel to the bottom. In the case of wires with a polygonal cross-section, the surfaces facing the bottom of the reaction vessels are inclined.

[0027] The predominantly inclined surfaces of the retention device ensure that during centrifugation, the centrifugal forces loosen and separate liquids adhering to the retention device due to adhesion forces and reliably remove them.

[0028] The retention device may further comprise a drainage device, wherein such a retention device is provided for a reaction vessel unit with a plurality of reaction vessels, each of which has a reaction vessel opening located in a common opening plane. The drainage device comprises a drainage surface configured such that, in a centrifuge, it is arranged opposite the reaction vessel openings of the reaction vessels and configured such that, during centrifugation, it rotates with the reaction vessel unit and, due to centrifugal acceleration, collects and drains liquid escaping from the reaction vessels in a controlled manner.

[0029] The discharge device is preferably inclined relative to the opening plane and, in particular, inclined such that, when the reaction vessel unit and the discharge device are arranged in the centrifuge, the discharge surface is inclined relative to the centrifuge's rotational axis. This ensures that any liquid escaping from the reaction vessels is discharged in a controlled manner in a direction parallel to the rotational axis.

[0030] Preferably, the discharge device is designed to have a concave discharge surface, which in particular is flush with two opposite edges of the reaction vessel unit. This defines a self-contained chamber between the reaction vessel unit and the discharge device, the atmosphere of which is entrained during rotation in the centrifuge.

[0031] The discharge device can be designed like the collecting devices known from WO 2023 / 148272 A1 and WO 2023 / 148273 A1.

[0032] According to a further aspect of the present invention, a method for automatically processing chemical or biological samples immobilized on a solid is provided.This method is characterized in that at least one solid body, on which one or more chemical or biological samples are immobilized, is arranged in at least one reaction vessel of a reaction vessel unit, wherein a retaining device is provided on the reaction vessel unit in such a way that an opening of the reaction vessel is covered by the retaining device in such a way that in the region of the opening of the reaction vessel at least one or more through openings are provided which are smaller than the reaction vessel opening and sufficiently small to retain the solid body in the reaction vessel during centrifugation with the reaction vessel opening directed radially outwards, wherein during centrifugation the solid body is separated from a liquid located in the reaction vessel.

[0033] In this method, the retention devices and / or the solid bodies (e.g. plastic ball, fabric, fleece, etc.) explained above can be used.

[0034] This process also allows specific liquids to be automatically dispensed into the respective reaction vessel and separated again by centrifugation. The liquids can remain in the respective reaction vessels for a specific period of time. The liquids or reaction vessels can be exposed to specific conditions while the liquid is in the reaction vessel, such as a specific temperature profile (e.g., for PCR amplification), exposure to specific radiation (light, X-ray particle radiation), or shaking.

[0035] The invention is explained in more detail below by way of example with reference to the drawings. The drawings show: Fig. 1a to 1f an embodiment of a restraint device in a plan view, isometric sectional view, isometrically sectioned exploded view, in a cross-sectional view and in an exploded view in a cross-section. Fig. 2a to 2f an embodiment of a restraint device in a plan view, isometric sectional view, isometrically sectioned exploded view, in a cross-sectional view and in an exploded view in a cross-section. Fig. 3a to 3f an embodiment of a restraint device in a plan view, isometric sectional view, isometrically sectioned exploded view, in a cross-sectional view and in an exploded view in a cross-section. Fig. 4a to 4f an embodiment of a restraint device in a plan view, isometric sectional view, isometrically sectioned exploded view, in a cross-sectional view and in an exploded view in a cross-section.

[0036] A first embodiment of a restraint device 1 ( Fig. 1a to 1f) has a disc-shaped and approximately flat base plate 2, which is formed centrally with a through opening 3.

[0037] The retaining device 1 is designed to be placed on a reaction vessel unit 4, which has a reaction vessel 5 ( Fig. 1c to 1f).

[0038] In the following, the area located on the side of the reaction vessel 5 with respect to the retaining device 1 is referred to as below or below the retaining device 1, and the area located on the other side of the retaining device 1 or on the other side of the base plate 2 is referred to as above or above with respect to the base plate 2. During use, particularly in a centrifuge, there are also situations in which the reaction vessels 5 can be located above the retaining device 1.

[0039] A tubular projection 6 extends downwards from the edge of the through-opening 3. At the end of the projection 6 remote from the base plate 2, a grid 7 is provided which extends over the entire cross-sectional area of ​​the tubular projection 6.

[0040] The tubular projection 6 has a downwardly tapered shape.

[0041] The reaction vessel 5 is formed from a bottom wall 8 and a circumferential shell wall 9. The shell wall 9 is shaped to widen conically upwards from the bottom wall 8, with the conical wall of the projection 6 of the retaining device 1 being shaped to complement the shell wall 9 in such a way that when the projection 6 is inserted into the reaction vessel 5, its circumferential wall engages the shell wall 9 with frictional engagement. This fixes the retaining device 1 to the reaction vessel unit 4.

[0042] Within the scope of the invention, it is of course also possible for the projection 6 to be designed such that it can be arranged with play in the respective reaction vessel 5. In such an embodiment, the retaining device 1 must be fixed in another way with respect to the reaction vessel unit 4.

[0043] In the Fig. 1c to 1f, a circumferential, disc-shaped collar 10 is shown on the upper edge of the reaction vessel 5. Preferably, the reaction vessel 5 is part of a reaction vessel unit, such as a microtiter plate, in which the collar 10 forms a flat plate on which a plurality of reaction vessels 5 are formed, each of which opens with its reaction vessel openings and is arranged in one plane. For simplicity of illustration, the present drawings only show reaction vessel units 4, each with a single reaction vessel 5. In practice, however, the reaction vessel units 4 generally comprise a plurality of reaction vessels 5 arranged in a regular grid, with the retaining device 1 then being correspondingly formed with a plurality of projections 6 arranged in the same grid.

[0044] The grid 7 is formed from several grid bars that are arranged at right angles to one another and thus define approximately square through-openings. The grid bars have a circular cross-section. They can also be designed with a cross-section in the shape of a polygon. Preferably, the grid bars are shaped such that their downward-facing surfaces run at an angle to a perpendicular that is arranged perpendicular to the plane of the base plate 2 or to the plane of the grid 7. When the retaining device 1 is used in a centrifuge, this perpendicular 11 essentially corresponds to the radial direction of the centrifuge. By arranging the downward-facing surfaces of the grid 7 at an angle with respect to the perpendicular 11, it is ensured that the centrifugal force can remove all of the liquid from the grid 7 and drive it radially outwards.

[0045] A second embodiment of the restraint device 1 again comprises a base plate 2 and a grid 7, wherein in this embodiment the grid 7 is arranged in the same plane as the base plate 2 and is fastened thereto ( Fig. 2a to 2f). At the edge of the through-opening 3 of the base plate 2, in which the grid 7 is located, a circumferential annular web 12 is formed on the base plate 2, so that when the retaining device 1 is placed on a reaction vessel unit 4, the annular web engages in a corresponding reaction vessel opening of the reaction vessel 5 and is fixed thereto by frictional engagement.

[0046] Both the annular web 12 of this second embodiment and the circumferential wall of the projection 6 of the first embodiment have an oblique lower edge 13 which is conically tapered from bottom to top so that during centrifugation liquid does not get stuck on the lower edge of the annular web 12 or the circumferential wall of the projection 6, but is reliably guided in the area of ​​the through-opening 3 of the base plate 2.

[0047] The grid 7 of the second embodiment is designed in the same way as the grid 7 of the first embodiment.

[0048] A third embodiment of the restraint device 1 ( Fig. 3a to 3f) is similar to the second embodiment ( Fig. 2a to 2f), wherein identical parts are designated by identical reference numerals and will not be explained again. This retaining device 1 also comprises a base plate 2 with a through-opening 3 and a grid 7 arranged in the same plane as the base plate 2 and attached thereto. The through-opening 3 of the base plate 2 has the same or a slightly larger opening width than the reaction vessel opening of the reaction vessel 5.

[0049] The reaction vessel 5 protrudes slightly from the collar 10 of the reaction vessel unit 4 with a circumferential annular web 14. The base plate 2 has a slightly downwardly protruding annular web 15, the inner width of which is dimensioned such that it rests positively on the outer edge of the annular web 14 of the reaction vessel unit 4. The two annular webs 14 and 15 position the retaining device 1 and the reaction vessel unit 4 precisely relative to one another. The frictional engagement between these two annular webs 14, 15 is so slight that it is expedient, during use, to fix the retaining device 1 to the reaction vessel unit 4 by other means. The retaining device 1 and the reaction vessel unit 4 can each be gripped behind by a rail of a centrifuge rotor at two opposite edge regions, so that both the retaining device 1 and the reaction vessel unit 4 are securely fixed to the rotor in the radial direction of the rotor.For example, the retaining device 1 and the reaction vessel unit 4 can be attached together to the rotor.

[0050] A fourth embodiment of the restraint device 1 ( Fig.4a to 4f) is designed similarly to the second and third embodiments, each having a base plate 2 with a through-opening 3 and a grid 7 arranged in the plane of the base plate. The fourth embodiment differs from the second and third embodiments in that the through-opening 3 and thus the grid 7 are somewhat larger than in the embodiments described above and can extend somewhat beyond the reaction vessel opening of the corresponding reaction vessel 5. With such a retaining device 5, it is possible to reliably cover differently sized reaction vessels 5 with the grid 7. Such a retaining device 1 is also suitable for reaction vessel units in which the reaction vessels have a square shape in plan view.

[0051] Within the scope of the invention, it is also possible to design the retaining device 1 such that a continuous grid is provided that extends over all reaction vessels 5 of a reaction vessel unit 4. Preferably, such a grid is provided at the edge region with a frame that has greater stability than the grid.

[0052] The restraint device 1 is preferably made of plastic. It can be formed as an injection-molded part. The grid struts are arranged in one plane.

[0053] In use, the retention device 1 is used to retain a solid, such as a plastic bead or a piece of fabric or nonwoven fabric, in the reaction vessel 5. The solid may come into contact with one or more liquids, which may be dispensed through the grid 7 into the reaction vessel 5. The liquid may be separated from the solid by centrifugation.

[0054] This can be done fully automatically using a suitable centrifuge with an integrated or separate dispensing unit. One such centrifuge is described, for example, in WO 2015 / 018878 A2.

[0055] The above-described embodiments can also be modified such that, instead of the grid 7, the solid body, for example a fabric, a fleece, or a film, is attached directly to the retaining device 1. This solid body can then be brought into contact with different liquids in the same way. REFERENCE SYMBOL 1 restraint device 2 base plate 3 passage opening 4 reaction vessel unit 5 Reaction vessel 6 lead 7 grids 8 Bottom wall 9 Shell wall 10 collars 11 lots 12 Ringsteg 13 sloping lower edge 14 Ringsteg 15 Ringsteg QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited patent literature

[0000] WO 2023 / 148272 A1 [0003, 0031] WO 2023 / 148273 A1 [0003, 0031] DE 20 2014 010 544 U1

[0004] DE 10 2021 124 023 A1

[0004] DE 10 2008 242 971 A1

[0005] WO 2023 / 110944 A1

[0006] WO 2015 / 018878 A2

[0054]

Claims

[1] Retaining device for a reaction vessel unit for centrifuging reagents in the reaction vessel unit, which has at least one blind-hole-shaped reaction vessel with a reaction vessel opening, wherein the retaining device is designed to be placed on or inserted into the at least one reaction vessel and has either one or more through-openings which are smaller than the reaction vessel opening, so that during centrifugation with the reaction vessel opening directed radially outwards, a solid body which is larger than the through-openings can be retained by the retaining device, or that at least one solid body for immobilizing one or more chemical or biological samples or which itself represents a sample is fastened to the retaining device in such a way that the solid body is located within the at least one reaction vessel when the retaining device is placed on a reaction vessel unit. [2] Restraint device according to claim 1, characterized by that the retaining device is designed to cover several reaction vessels of the reaction vessel unit. [3] Restraint device according to claim 1, characterized by that the retaining device is designed as an insert element for insertion into the at least one reaction vessel. [4] Restraint device according to one of claims 1 to 3, characterized by that the retaining device is designed with a complementary shape to at least a portion of the at least one reaction vessel. [5] Restraint device according to one of claims 1 to 4, characterized by that the restraint device is designed as a grid with wire-shaped grid struts. [6] Restraint device according to claim 5, characterized by that the grid struts have a maximum thickness of 1 mm, preferably 0.5 mm and in particular 0.3 mm. [7] Restraint device according to one of claims 1 to 6, characterized by that the surfaces facing a bottom of the at least one reaction vessel unit are predominantly inclined to a horizontal extension of the reaction vessel unit. [8] Restraint device according to one of claims 1 to 7, characterized by in that the retention device has a drainage device, wherein the reaction vessel unit has a plurality of reaction vessels, each with a reaction vessel opening, which lie in a common opening plane, wherein the drainage device has a drainage surface which is designed such that it is arranged opposite the reaction vessel openings of the reaction vessels in a centrifuge, and is designed such that it rotates with the reaction vessel unit during centrifugation and catches and drains liquid emerging from the reaction vessels due to the centrifugal acceleration.

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