Mixing device and method for providing a mixture

Abstract

The invention relates to a method for providing a mixture from an interior space (17) of a container (1), wherein a content (14, 15) in the interior space (17) comprises a liquid (14) and a constituent (15), and the container (1) comprises at least one opening (2) for the intake and / or withdrawal of the content (14, 15), wherein a supply element (5) is arranged at the container (1) such that a fluid can be supplied by means of the supply element (5) into the interior space (17) of the container (1), wherein the method comprises the following steps: supplying the fluid by means of the supply element (5) into the liquid (14) in the interior space (17), wherein the liquid (14) is set in motion by the addition of the fluid such that the liquid (14) and the constituent (15) are mixed, causing the constituent (15) to be dispersed in the liquid (14); and providing the mixture by withdrawing a specifiable amount of the content (14, 15) from the interior space (17) as the fluid is being supplied.

Description

[0001] Hombrechtikon Systems Engineering AG, Garstliqweq 6, CH-8634

[0002] Hombrechtikon

[0003] Mixing device and method for providing a mixture

[0004] The invention relates to a mixing device and a method for providing a mixture according to the preamble of the independent claims.

[0005] Many methods for purifying and processing cells, DNA, and other biomolecules are known in the art. One type of purification is DNA extraction, in which the DNA is precipitated in a nonpolar environment. DNA can also be purified by centrifugation, e.g., after cell disruption, or by electrophoretic methods.

[0006] Biomolecules can also be synthesized and purified by immobilizing them on an insoluble support. Common substrates for immobilizing biomolecules include glass, as well as other, less common substrates such as gold, platinum, oxides, semiconductors, and various polymer substrates.

[0007] Since manual purification and processing in numerous steps is too time-consuming, these processes are now fully automated. Magnetic beads (magnetic particles) play a key role in the automation of laboratory methods.

[0008] Magnetic bead-based cleanup and magnetic bead-based normalization are widely used methods for the immobilization, purification, and concentration adjustment of nucleic acids. Typical applications of these methods include sample preparation in the context of DNA sequencing or DNA detection (e.g., using PCR, or polymerase chain reaction). The magnetic particles were developed in 1995 at the Whitehead Institute for the purification of PCR products. The magnetic particles are usually paramagnetic and can consist, for example, of polystyrene coated with iron. Various molecules with carboxyl groups can then be attached to the iron. These carboxyl groups can reversibly bind the DNA molecules, thus immobilizing the DNA molecules.

[0009] Methods using magnetic particles, for example, include the following steps: First, the PCR products are bound to the magnetic particles.

[0010] This typically involves mixing the liquid and the magnetic particles it contains. The magnetic particles, along with the attached PCR products, are then separated from impurities (this step can be achieved, for example, by pipetting the solution away from the solid / container). The magnetic particles, along with the attached PCR products, are then washed. After washing, the PCR products are eluted from the magnetic particles and transferred to a new plate.

[0011] In fully automated processes, the necessary reagents are automatically pipetted into the sample after the starting material has been introduced in an isolation process and then removed again. The magnetic particle-bound nucleic acids are collected at the bottom and edges of the containers and, depending on the routine, are redissolved by optimized pipetting. Finally, the DNA or RNA is eluted into separate, capped containers for immediate storage or further use.

[0012] One of the most important methods for synthesizing, normalizing, and purifying biomolecules is the method using magnetic particles. In this method, the biomolecules are bound to the surface of the magnetic particles. The magnetic particles are then immobilized using a magnet, and the solution, which contains byproducts and impurities, can be easily separated. This allows the biomolecules to be purified and isolated quickly and easily. The advantage of magnetic beads is that the beads can move freely within the experimental setup, which is important for the binding steps. If you want to remove the liquid from the container, for example, in a washing step, you simply hold a magnet to the container and the liquid can then be separated.

[0013] The magnetic particles are small paramagnetic or ferromagnetic spheres coated with various materials that impart the required properties. Nickel particles coated with a plastic are often used.

[0014] In the current state of the art, the aforementioned steps are usually carried out in bioreactors, in which the magnetic particles and the liquid are mixed with biomolecules to ensure the most complete binding of the biomolecules to the magnetic particles. However, such bioreactors or mixing devices can also be used for the processing and mixing of other biological or chemical substances, such as cells.

[0015] A corresponding mixing device is known from US 2022 / 0135924 A1. The mixing device of US 2022 / 0135924 A1 is intended to provide optimal conditions for mixing a reaction liquid. The mixing is based on a change in the internal volume of the reaction vessel, which triggers movement of the liquid. The change in the internal volume is based in particular on a change in the shape of the reaction vessel, for which the reaction vessel has at least one flexible area / region. The flexible area is a flexible base, which is moved by a magnetic drive element connected to the base. This creates a mixing movement. However, such mixing devices do not ensure efficient or complete mixing. Furthermore, the flexible areas of the vessel represent a weak point in the material.It is known from the state of the art that materials that are frequently stretched and compressed and exposed to changing temperatures will fatigue over time, which can lead to fluid leakage.

[0016] The object of the invention is therefore to provide a mixing device and a method for using a mixing device that avoid the adverse effects known from the prior art, in particular enabling rapid, complete, and efficient mixing. Specifically, it should be possible to ensure that a reproducible concentration / amount of, in particular, cells, magnetic particles, molecules, or other particles or components is always present in a withdrawn volume of liquid, while preventing any leakage of liquid.

[0017] The object is achieved by a mixing device and by a method for providing a mixture having the features of the independent claims.

[0018] According to the invention, a method for providing a mixture from the interior of a container is proposed. The contents in the interior comprise a liquid and a (particularly solid) component, and the container has at least one opening for receiving and / or removing the contents. In addition, a feed element is arranged on the container so that a fluid can be fed into the interior of the container via the feed element. The method according to the invention comprises the following steps: The fluid is fed into the liquid in the interior via the feed element, wherein the liquid is set in motion by the addition of the fluid such that the liquid and the component are mixed, so that the component is (particularly evenly) distributed in the liquid. The mixture is provided by removing a predeterminable amount of the contents from the interior while the fluid is being fed.

[0019] By supplying the fluid to the liquid in the interior of the container, a movement of the contents, in particular the liquid, is generated, thus inducing a mixing movement / flow of the liquid, which leads to mixing. Thus, by supplying the fluid, the flow in the container is generated, which results in a homogeneous mixing of the contents. By supplying the fluid, the contents of the container can be mixed flexibly and efficiently, whereby the mixing can be easily adjusted by adjusting the amount of fluid supplied.

[0020] The mixture can be used for further chemical, biochemical, and / or biological processes and / or process steps; in particular, the mixture can be transferred to another container for this purpose. Preferably, the mixture is removed from the interior of the container using an instrument for removing the liquid, such as a pipette or syringe.

[0021] In this case, the instrument can therefore comprise the pipette or the syringe, which can in particular also be designed as a robot and can comprise a plurality of pipettes and / or syringes.

[0022] In the embodiment of the invention, the component may, for example, comprise cells (or a plurality of them), in particular consist of cells. Alternatively or additionally, the liquid may comprise biomolecules, and the liquid may be a suitable solvent, in particular comprising water.

[0023] Alternatively or additionally, the component may comprise (or a plurality of) magnetic particles, in particular consisting of magnetic particles. The method according to the invention may comprise collecting the magnetic particles on a wall of the container using a magnet. Furthermore, the collected magnetic particles may be resuspended. In particular, these steps may be performed by moving the magnet. In practice, a molecule (such as biomolecules) in the liquid may bind to the magnetic particles. The magnetic particles and the molecules bound to them may be transferred to a second container, and / or the magnetic particles and the molecules bound to them may be separated from one another, and / or the biomolecules may be enriched using an elution buffer and a washing step.

[0024] The fluid is preferably a gas, wherein the gas particularly comprises CO2 and / or oxygen and / or an inert gas and / or air. If the fluid is a gas, this has the advantage that the volume of the liquid in the interior and the concentration of the component are not changed by the addition of the fluid. The gas can be introduced at a bottom / underside / lower region of the container in such a way that, upon rising in the liquid, the gas sets the liquid in motion in such a way that the liquid and the component are mixed, and the component is distributed throughout the liquid.

[0025] In practice, the supply of the fluid can be controlled via a regulating element and the fluid can be supplied from a source (such as a gas container or a gas cylinder) via a conduit element (such as a pipe / pipeline) to the container.

[0026] The fluid can be in a liquid state upon introduction, in particular a liquid. Thus, by introducing the fluid, the concentration of the component can be adjusted, while simultaneously mixing and adjusting the concentration. Alternatively, the fluid can be introduced in a liquid state and then transition to a gaseous state in the liquid. Thus, in addition to mixing, a cooling effect can be created, for example, by using liquid nitrogen.

[0027] According to the invention, a mixing device for use in the method according to the invention is further proposed. Here, the mixing device comprises the container with the interior for receiving the contents, wherein the container comprises the opening for receiving and / or removing the contents and a connecting element separate from the opening. The container is replaceable, i.e., can be removed from the mixing device and replaced with another container. For this purpose, the mixing device comprises a housing with a feed element for feeding the fluid into the interior of the container and a receiving element for receiving the container. Furthermore, the container is arranged in the receiving element, and the feed element can be arranged on the connecting element such that the fluid can be fed into the interior of the container.

[0028] In practice, the mixing device can comprise a plurality of feed elements and / or connecting elements (which are arranged in particular at different locations on the container / evenly distributed on the container) to achieve more flexible control and more thorough mixing. In an embodiment of the invention, the feed element can comprise a regulating element for adjusting a flow rate of the fluid, wherein the regulating element can be and / or comprise a nozzle and / or a valve (such as a lip check valve). The regulating element, as well as the other device parts and method steps, can be controlled via a control unit. The mixing device can comprise the control unit.

[0029] In a preferred embodiment, the container can be supported on a support surface of the receiving element, and the feed element can comprise a fixing element. The feed element can be displaceable relative to the receiving element, so that the container can be fixed between the fixing element and the support surface after being arranged in the receiving element. The mixing device can comprise a movement device which can be connected to the feed element or is part of the feed element, so that the feed element can be moved by the movement device. For this purpose, the movement device can comprise a drive, in particular a motor (such as an electric motor, in particular a servo motor). The feed element can comprise a rail, so that a movement of the feed element (and thus also a movement distance) is predetermined by the rail.Alternatively, the feed element can be arranged on an element / support / holder which is moved by the drive.

[0030] Preferably, the connecting element is a connection opening separate from the opening, and the feed element comprises a feed tube, which feed tube can be inserted / extended into the connection opening such that the fluid can be fed into the interior of the container. For this purpose, the connection opening can comprise a sealing element which seals the connection opening and into which the feed tube can be inserted. In particular, the sealing element can be designed in the form of a membrane (which can be made of a polymer / plastic, for example), by which the opening is closed. The feed tube can be passed through / pierced through the membrane such that the fluid can be fed into the interior of the container. Preferably, the feed tube is designed as a needle / needle-shaped for piercing. In addition, the opening can also be provided with such a sealing element / membrane.Alternatively or additionally, the mixing device can comprise the source connected to the supply element via the conduit element, wherein the fluid can be conducted from the source to the supply element via the conduit element. In practice, the container can be designed, in particular, as a consumable, i.e., as a consumable material that is introduced into the mixing device and can be disposed of after a corresponding process or method has been performed. A new container can then be introduced into the mixing device. The container is therefore replaceable.

[0031] The container according to the invention is particularly preferably chemically resistant, pH-stable, and temperature-resistant. Furthermore, the container can be designed to hold at least 10 ml of liquid / contents, preferably 10 ml to 500 ml or 20 ml to 100 ml, particularly preferably 20 ml to 50 ml of liquid / contents. The container can, in particular, comprise a polymer, particularly preferably consist of a polymer. Examples of polymers are polyethylene, polypropylene, polyethylene terephthalate, and / or polyetheretherketone.

[0032] The interior of the container can contain a liquid and, as components, a large number of magnetic particles / cells / molecules, other particles or parts that sink in a liquid. The liquid and the large number of magnetic particles / cells / molecules, other particles or sinking parts can be mixed by introducing the fluid. A pre-filled container of this type can in particular be provided and sold as a consumable. The mixability ensures in particular that the concentration of magnetic particles / cells / molecules, other particles or sinking parts is the same for each volume of liquid / contents (i.e. the mixture / specified amount) withdrawn, which can then be used in further steps or methods for processing, for example, biomolecules.The mixing device / container can in particular be a mixing device / container for providing a liquid with magnetic particles / cells / molecules, other particles, or sinking parts. In one embodiment, the mixing device can further comprise the magnet, wherein the magnet can be arranged on the container in such a way that magnetic particles that can be introduced into the interior can be fixed in the container. This allows the liquid to be drained off and a new liquid to be added without removing the magnetic particles from the interior. The magnet can be a permanent magnet and / or an electromagnet. To drain (or add) the liquid, the instrument for removing the liquid, such as a pipette, can be used so that the liquid can be removed from the container after molecules have been immobilized on the surface of the magnetic particles.This can be done through the opening of the container. The instrument used to remove the fluid can also be a syringe or another suitable instrument.

[0033] In practice, the container and mixing device according to the invention are preferably used to provide liquid containing magnetic particles for other processes, such as biomolecule processing (whereby a consistent / reproducible concentration of magnetic particles can be provided for each withdrawn volume of liquid). Alternatively, the container according to the invention can be a reaction vessel and the mixing device a bioreactor in which biomolecules are processed with magnetic particles.

[0034] When used as a reaction container, the magnet can be movably mounted on the container / in the mixing device such that the magnetic particles are freely movable within the container during a reaction step and are fixed in place during a washing step by changing the magnet position. In particular, the magnet can be movable such that the magnet is arranged in a first position on the container, fixing the magnetic particles, and moving the magnet to a second position makes the magnetic particles movable. Of course, in an automated device, for example, the container can also be moved relative to the magnet to achieve the same effect.

[0035] In the method according to the invention, the contents can also optionally be introduced into the interior of the container first. The contents can, for example, comprise magnetic particles and the liquid, and the magnetic particles and the liquid can be mixed in the interior by adding the fluid, thus preventing the magnetic particles from sinking.

[0036] In particular, the method can be a method for providing a liquid with magnetic particles / cells / molecules, other particles, or sinking particles, in which magnetic particles / cells / molecules, other particles, or sinking particles and the liquid (as contents) are introduced into the interior of the mixing device, and the liquid and magnetic particles / cells / molecules, other particles, or sinking particles are mixed in the interior. A predeterminable volume of liquid / contents can be withdrawn from the container, particularly preferably while the mixing / addition of the fluid is taking place.

[0037] Additionally, the method may be a method for processing biomolecules, in which magnetic particles and a liquid containing biomolecules (as contents) are introduced into the interior of the mixing device, and the liquid and magnetic particles are mixed in the interior. Furthermore, the method may include securing the magnetic particles in the container by arranging a magnet on the container and / or removing the liquid containing magnetic particles from the interior using the liquid removal instrument.

[0038] In the method according to the invention, in particular, the magnetic particles (also magnetic particles) / cells / molecules, other particles or sinking parts and the liquid are mixed by the action of the fluid on the contents in order to obtain a liquid with magnetic particles / cells / molecules, other particles or sinking components, which has a uniform concentration of magnetic particles / cells / molecules, other particles or sinking parts.

[0039] Additionally or alternatively, the supply of the fluid is designed to keep the particles, cells, molecules, or sinking particles contained in the interior suspended in such a way that sedimentation at the bottom of the container can be prevented. Furthermore, a mixing or swirling process is improved by keeping the particles suspended in the vessel and / or by preventing or reducing coagulation of beads. Allowing the magnetic particles to float freely and / or preventing sedimentation of the magnetic particles advantageously improves biochemical reactions in the mixing device, namely biomolecule immobilization.

[0040] In the context of the present invention, the term biomolecule can be understood as DNA, RNA, nucleic acids, proteins, start sequences, monomers, or other biologically active molecules. Such biomolecules can be processed in the mixing device according to the invention and introduced into the container according to the invention (preferably in a liquid). The processing can include, among other things, washing steps and / or reaction steps and can be carried out with magnetic particles. A washing step in the context of the invention is a process step in which the liquid is removed from the container after thorough mixing, and in particular the impurities are separated from the magnetic particles with the attached biomolecules. A washing step can also include rinsing with a rinsing fluid.Within the scope of the invention, a reaction step is a process step in which the biomolecules bound to the magnetic particles are converted, bound to the particles, or extended (chain extension, e.g., PCR "polymerase chain reaction"). A contaminant is understood in particular to be a substance that has not fully reacted or is not bound to the magnetic particles, the solvent, byproducts, and contaminants, as well as a mixture of two or more of the above-described substances.

[0041] Within the scope of the invention, a liquid or the content can be a solution or suspension, in particular a mixture of liquid and magnetic particles or a reaction mixture of biomolecules and / or reagents and / or impurities.

[0042] For the purposes of the invention, “magnetic particle” (also “magnetic bead”) can be understood to mean a particle in the micrometer or millimeter range. Furthermore, the magnetic particles can be porous. A biomolecule can be bound to the surface of the magnetic particles in particular via thiol groups and / or amino groups and / or hydroxyl groups and / or carboxyl groups and / or carbonyl groups and / or ester groups and / or nitrile groups and / or amine groups and / or any other functional groups or other interactions. Magnetic particles can also be coated nickel particles or any other ferro- or paramagnetic particles. Magnetic particles typically have a diameter of approximately 1 micrometer. For the purposes of the invention, approximately 1 micrometer is understood to mean 0.5 to 1.5 micrometers, in particular 0.7 to 1.3 micrometers, particularly preferably 0.9 to 1.1 micrometers.

[0043] The aforementioned magnetic particles can therefore be functionalized magnetic particles that bind biomolecules such as DNA in a sample material. They can then be immobilized using magnetic force and thus separated from a reagent liquid. This is removed, and the biomolecules can then be detached from the particles. The magnetic particles should preferably be added in a precise amount. This is made possible by the devices and method according to the invention. The mixing device can be used for the preparation of or in any enzymatic process involving nucleic acids (e.g., polymerase chain reaction (PCR), isothermal DNA amplification, reverse transcription). The biomolecules bound to the magnetic particles can be detached from the surface of the magnetic particles and subsequently reused.

[0044] In the following, the invention is explained in more detail using exemplary embodiments with reference to the drawings.

[0045] Fig. 1 is a sectional view of a first embodiment of a mixing device according to the invention;

[0046] Fig. 2A is a plan view of a receiving element with a container according to the invention;

[0047] Fig. 2B is a side view of the receiving element according to the invention with container according to Fig. 2A;

[0048] Fig. 3 is a schematic representation of a second embodiment of a mixing device according to the invention;

[0049] Fig. 4 is a schematic representation of a mixing device with a pipette according to the invention.

[0050] A mixing device is, in particular, a device with a container in which liquids containing magnetic particles can be provided under adjustable and thus optimal conditions. The operation of a mixing device is therefore, in particular, an application of chemistry, biochemistry, or biotechnology, which enables chemical and biological processes or provides starting materials for them. Factors that can be controlled or monitored in mixing devices include the composition of the contents, the gas supply, temperature, pH, sterility, and others. The mixing device can be used to provide a liquid with a predefined concentration of magnetic particles / cells / molecules / other particles and / or sinking particles, or for the extraction and processing of biomolecules. The processing and degradation of chemical compounds can also take place in mixing devices.

[0051] Fig. 1 shows a sectional view of a first embodiment of a mixing device 11 according to the invention for use in the method according to the invention.

[0052] The mixing device 11 comprises a housing 4 and a container 1 which has an interior space 17. This interior space 17 is suitable for accommodating a content such as a liquid and a component. The container 1 further comprises an opening 2. Through this opening 2, the content can be removed from the interior space 17 of the container 1 using a pipette, or another substance can be added to the content of the interior space 17. The container 1 can be connected to a feed element 5 via a connecting element 3 which is different from the opening. A fluid can be fed into the interior space 17 of the container 1 via this feed element 5, which can be arranged on or in the housing 4. In the present exemplary embodiment, this takes place via a side chamber 31 which is connected to the interior space 17 via a side opening 32 at the bottom of the container 1.

[0053] In addition, the housing 4 comprises a receiving element 6 that can accommodate the container 1. The container 1 is arranged in the receiving element 6 in such a way that the feed element 5 can be arranged on the connecting element 3 (by inserting a feed tube 18 into the connecting element 3) so that the fluid can be guided into the interior 17 of the container 1. By introducing this fluid into the container 1, the liquid already contained therein is set in motion. This leads to the liquid and the component also located in the container 1 being mixed, thus achieving a uniform concentration of the component in the liquid.

[0054] The mixing device 11 shown here also has a source 13, which is also arranged in or on the housing 4 and is connected to the feed element 5 by means of a line element 12, so that the fluid can pass from the source 13 via the line element 12 to the feed element 5 and from there into the container 1.

[0055] Furthermore, the receiving element 6 comprises a support surface 8, wherein the container 1 is supported on the support surface 8 of the receiving element 6. The feed element 5 comprises a fixing element 9, wherein the feed element 5 is displaceable relative to the receiving element 6, such that the container 1, after being arranged in the receiving element 6, can be fixed between the fixing element 9 and the support surface 8 by moving the fixing element 9 over the container 1. Since the container 1 is arranged on a spring 20 via the support surface 8 (or via a support element), the container 1 can be clamped between the fixing element 9 and the support surface 8, since the spring 20 can exert pressure on the container 1 located in the receiving element 6.

[0056] Fig. 2A shows a plan view of a receiving element 6 according to the invention with container 1 .

[0057] The container 1 is arranged in the receiving element 6 such that the receiving element 6 surrounds the container 1. The interior 17 of the container 1 can be accessed from above via the opening 2. The fixing element 9, which can be pushed over a wall 19 of the container 1 and thus fixes the container 1 in the receiving element 6. The fixing element 9 is arranged movably in the housing 4 / on the receiving element 6 such that the container 1 can be inserted into the receiving element 6 and is fixed in the receiving element 6 by moving the fixing element 9 over the wall 19.

[0058] Fig. 2B shows a side view of the receiving element 6 with the container 1. The support surface 8 is arranged on a platform 22 by a cylindrically shaped pressure transmission element 80, which in turn is arranged by the spring 20 in such a way that pressure can be exerted on the container 1 located in the receiving element 6 by means of the spring 20 via the platform 22 and the support surface 8.

[0059] Fig. 3 shows a schematic representation of a second embodiment of a mixing device 11 according to the invention.

[0060] The interior 17 of the container 1 comprises a liquid 14 and the component 15 located therein. The container 1 comprises the opening 2 for receiving and / or removing the contents 14, 15, wherein a feed element 5 is arranged on a lower side of the container 1, so that a gas can be fed via the feed element 5 through the connecting element 3 into the interior 17 of the container 1.

[0061] The gas is fed along direction A via the feed element 5 into the liquid 14 in the interior space 17, wherein the liquid 14 is set in motion by the addition of the gas in such a way that the liquid 14 and the component 15 are mixed, so that the component 15 is distributed in the liquid 14.

[0062] In particular, the gas is introduced at the bottom of the container 1 in such a way that, when the gas rises in the liquid 14, the liquid 14 is set in motion in such a way that the liquid 14 and the component 15 are mixed.

[0063] A predeterminable amount of the contents 14, 15 can be removed from the interior 17 via the opening 2 during the supply of the gas.

[0064] Fig. 4 shows a schematic representation of a section of a mixing device 11 according to the invention with the pipette 16, wherein, among other things, the feed element is not explicitly shown.

[0065] In this example, the pipette 16 can be guided through the opening 2 of the container 1 into the interior 17 and immersed into the liquid 14 containing the components 15. By actuating the pipette 16, the liquid 14 and component 15, which is suspended in the liquid by a fluid flow, can be picked up and removed from the container 1 through the opening 2. Likewise, liquid 14 and / or component 15 can be supplied into the interior 17 of the container 1 through this opening 2.

[0066] In the present embodiment, the component 15 is a plurality of magnetic particles 15. The mixing device 11 comprises the magnet 7, wherein the magnet 7 is arranged on the container 1 in such a way that the magnetic particles 15 located in the container 1 are fixed. This allows the liquid 14 to be discharged by means of a pipette 16 without magnetic particles 15 after the pipette 16 has been inserted through the opening 2 into the interior 17 of the container 1. If the magnet 7 is moved away from the container 1, the liquid 14 with a predeterminable concentration of magnetic particles 15 can be withdrawn.

[0067] While the invention has been shown and described in detail in the drawings and foregoing description, such illustration and description are to be regarded as illustrative or exemplary and not restrictive.

[0068] The invention is not limited to the disclosed embodiments.

[0069] Other variations of the disclosed embodiments may be understood and effected by those skilled in the art in practicing a claimed invention from a study of the drawings, the disclosure, and the dependent claims. In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite article "a" or "an" does not exclude a plurality. The mere fact that certain measures are repeated in mutually different dependent claims does not mean that a combination of those measures cannot be advantageously used. Any reference signs in the claims should not be construed as limiting the scope.

Claims

1. A method for providing a mixture from an interior space (17) of a container (1), wherein a content (14, 15) in the interior space (17) comprises a liquid (14) and a component (15), and the container (1) comprises at least one opening (2) for receiving and / or removing the content (14, 15), wherein a feed element (5) is arranged on the container (1) so that a fluid can be fed into the interior space (17) of the container (1) via the feed element (5), the method comprising the following steps: a) feeding the fluid into the liquid via the feed element (5) (14) in the interior (17), wherein the liquid (14) is set in motion by the addition of the fluid in such a way that the liquid (14) and the component (15) are mixed, so that the component (15) is distributed in the liquid (14), b) providing the mixture by removing a predeterminable amount of the contents (14, 15) from the interior (17) during the supply of the fluid.

2. The method according to claim 1, wherein the component (15) comprises cells, in particular consists of cells.

3. Method according to one of the preceding claims, wherein the liquid (14) comprises biomolecules.

4. Method according to one of the preceding claims, wherein the component (15) comprises magnetic particles (15), in particular consists of magnetic particles (15).

5. The method according to claim 4, wherein the magnetic particles (15) are collected by a magnet (7) on a wall of the container (1).

6. The method according to claim 5, wherein the collected magnetic particles (15) are resuspended.

7. Method according to one of claims 4 to 6, wherein a molecule in the liquid (14) binds to the magnetic particles (15).

8. The method according to claim 7, wherein the magnetic particles (15) and the molecules bound thereto are transferred into a second container.

9. The method according to claim 8, wherein the magnetic particles (15) and the molecules bound thereto are separated from each other.

10. The method according to claim 9, wherein the biomolecules are enriched by an elution buffer and a washing step.

11. A method according to any one of the preceding claims, wherein the mixture is transferred to another container.

12. Method according to one of the preceding claims, wherein the mixture is removed from the interior (17) of the container (1) using a pipette (16) or a syringe.

13. Method according to one of the preceding claims, wherein the fluid is a gas, in particular CO2 and / or oxygen and / or an inert gas.

14. The method according to claim 13, wherein the gas is introduced at a bottom and / or a bottom of the container (1) in such a way that the gas, when rising in the liquid (14), sets the liquid (14) in motion in such a way that the liquid (14) and the component (15) are mixed and the component (15) is distributed in the liquid (14).

15. Method according to one of the preceding claims, wherein a supply of the fluid is controlled via a regulating element.

16. Method according to one of the preceding claims, wherein the fluid is supplied from a source (13) via a conduit element (12) to the container (1).

17. Method according to one of the preceding claims, wherein the fluid has a liquid aggregate state when supplied, in particular is the liquid (14).

18. Mixing device for use in a method according to one of the preceding claims, comprising an exchangeable container (1) with an interior space (17) for receiving a content (14, 15), wherein the container (1) comprises an opening (2) for receiving and / or removing the content (14, 15) and a connecting element (3) separate from the opening (2); and a housing (4) with a feed element (5) for feeding a fluid into the interior space (17) of the container (2) and a receiving element (6) for receiving the container (1), wherein the container (1) is arranged in the receiving element (6) and the feed element (5) can be arranged on the connecting element (3) such that the fluid can be fed into the interior space (17) of the container (1).

19. Mixing device according to claim 18, wherein the feed element (5) comprises a regulating element for adjusting a flow rate of the fluid.

20. Mixing device according to claim 18 or 19, wherein the container (1) is supported on a support surface (8) of the receiving element (6) and the feed element (5) comprises a fixing element (9), wherein the feed element (5) is displaceable relative to the receiving element (6) so that the container (1) can be fixed between the fixing element (9) and the support surface (8) after being arranged in the receiving element (6).

21. Mixing device according to one of claims 18 to 20, wherein the connecting element (3) is a connecting opening (3) separate from the opening (2) and the feed element (5) comprises a feed pipe (11), which feed pipe (18) can be introduced into the connecting opening (3) in such a way that the fluid can be fed into the interior (17) of the container (1).

22. Mixing device according to claim 21, wherein the connecting opening (3) is closed by a membrane and the feed pipe (18) can be passed through the membrane in such a way that the fluid can be fed into the interior (17) of the container (1).

23. Mixing device according to one of claims 18 to 22 comprising a source (13) connected to the feed element (5) via a line element (12), wherein the fluid can be conducted from the source (13) via the line element (12) to the feed element (5).

24. Mixing device according to claim 19, wherein the regulating element comprises a nozzle and / or a valve.

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