Mixing apparatus and method for providing a mixture

Abstract

The present invention relates to a method for providing a mixture from the interior 17 of a container 1, wherein the contents 14, 15 in the interior 17 comprise a liquid 14 and a component 15, the container 1 comprises at least one opening 2 for receiving and / or removing the contents 14, 15, and a supply element 5 is positioned in the container 1 so that a fluid can be supplied into the interior 17 of the container 1 via the supply element 5, the method comprising the steps of supplying a fluid into the liquid 14 in the interior 17 via the supply element 5, the liquid 14 moving in such a manner that the fluid is added so that the liquid 14 and the component 15 are mixed and as a result the component 15 is dispersed in the liquid 14, and providing a mixture by removing a predetermined amount of the contents 14, 15 from the interior 17 while supplying the fluid.

Description

Technical Field

[0001] The present invention relates to a mixing device and a method for providing a mixture as described in the preamble of the independent claim.

Background Art

[0002] In the state of the art, many techniques for purifying and processing cells, DNA, and other biomolecules are known. One method of purification is DNA extraction in which DNA is precipitated in a nonpolar environment. DNA can also be purified, for example, by centrifugation or by electrophoresis techniques after cell lysis.

[0003] Biomolecules can also be synthesized and purified by immobilization on an insoluble support. Common substrates for immobilizing biomolecules are glass as well as other less common substrates such as gold, platinum, oxides, semiconductors, and various polymer substrates.

[0004] Manual purification and processing require a great deal of time for many operations, so those processes are now fully automated. Therefore, so-called "magnetic beads" (magnetic particles / magnetite particles) play an important role in the automation of experimental techniques.

[0005] "Magnetic bead-based cleaning" and "magnetic bead-based normalization" are methods widely used in the immobilization, purification, and adjustment of nucleic acid concentrations. Typical applications of those methods are the preparation of samples in the context of DNA sequencing or DNA detection (for example, using PCR, English: polymerase chain reaction, German: Polymerase Kettenreaktion).

[0006] Magnetic particles were developed at the Whitehead Institute in 1995 for the purification of PCR products. Magnetic particles are typically paramagnetic and can be made, for example, from iron-coated polystyrene. Different molecules containing carboxyl groups can then be attached to the iron. These carboxyl groups can reversibly bind DNA molecules, thereby immobilizing them.

[0007] A method using magnetic particles includes, for example, the following steps: First, the PCR product is bound to the magnetic particles. This process generally involves mixing the liquid with the magnetic particles placed in the liquid. Next, the magnetic particles with the attached PCR product are separated from contaminants (this step is achieved, for example, by pipetting the solution from a solid / container). Then, the magnetic particles with the attached PCR product are washed. After washing, the PCR product is eluted from the magnetic particles and transferred to a new plate.

[0008] In a fully automated process, the required reagents are automatically pipetted as samples and removed again in the separation process after the substrate is introduced. Magnetic particle-bound nucleic acids are collected at the bottom and rim of the container and introduced back into the solution by optimizing intermittent pipetting. Finally, the DNA or RNA is eluted into separate containers with lids for immediate storage for further use.

[0009] Therefore, one of the most important methods for the synthesis, normalization, and purification of biomolecules is the method using magnetic particles. In this specification, biomolecules are bound to the surface of magnetic particles. The magnetic particles are then attached by a magnet, and the solution containing by-products and contaminants can be easily separated. In this way, biomolecules can be easily purified and detached. The advantage of magnetic beads is that the beads can move freely in a trial batch, which is important for the binding step. Now, for example, if we want to remove liquid from a container in the washing step, we can simply hold a magnet in the container and then separate the liquid.

[0010] Magnetic particles are small paramagnetic or ferromagnetic beads coated with different materials to give them the desired properties. Nickel particles coated with plastic are often used.

[0011] In current technology, the steps described above are generally performed in a bioreactor, where mixing of magnetic particles and a liquid containing biomolecules is carried out to ensure that the biomolecules bind to the magnetic particles as completely as possible. However, such bioreactors or mixing devices can also be used for processing and mixing other biological or chemical substances, such as cells.

[0012] A corresponding mixing apparatus is known from U.S. Patent Application Publication No. 2022 / 0135924(A1). The mixing apparatus of U.S. Patent Application Publication No. 2022 / 0135924(A1) provides optimal conditions for mixing reaction liquids. Mixing in this case is based on a change in the internal volume of the reaction vessel, which is caused by the movement of fluid. The change in internal volume depends particularly on a change in the shape of the reaction vessel, and for this purpose the reaction vessel has at least one flexible region / one flexible area. The flexible region in this case is a flexible bottom, which is moved by a magnetic drive element connected to the bottom. This causes a mixing motion. However, such a mixing apparatus does not guarantee efficient or complete mixing. Furthermore, the flexible region of the vessel introduces a weakness in the material. From the current art, it is known that materials that are frequently stretched and compressed and exposed to changing temperatures will eventually fatigue, and as a result, leakage of liquid may occur. [Prior art documents] [Patent Documents]

[0013] [Patent Document 1] U.S. Patent Application Publication No. 2022 / 0135924(A1) [Overview of the project] [Problems that the invention aims to solve]

[0014] Therefore, an object of the present invention is to provide a mixing apparatus and a method of using the mixing apparatus that avoid the adverse effects known from the existing art, particularly enabling high-speed, complete, and efficient mixing. In particular, it should be possible to prevent leakage of the liquid while ensuring that a reproducible concentration / amount of cells, magnetic particles, molecules, or other particles or components is always present in the volume of the removed liquid. [Means for solving the problem]

[0015] This objective is satisfied by mixing apparatus and methods for providing a mixture having a characteristic or independent claim.

[0016] According to the present invention, a method for providing a mixture from inside a container is proposed. Hereinafter, the contents inside comprise a liquid and (particularly solid) components, and the container has at least one opening for receiving and / or removing the contents. Furthermore, a supply element is positioned in the container so that a fluid can be supplied into the interior of the container via the supply element. The method according to the present invention comprises the following steps: A fluid is supplied into the liquid inside via the supply element, and the liquid is moved in such a manner that the liquid and components are mixed by the addition of the fluid, resulting in the components being dispersed (particularly evenly) in the liquid. Hereinafter, the mixture is provided by removing a predetermined amount of the contents from inside while supplying the fluid.

[0017] By supplying a fluid into the liquid inside a container, movement of the contents, particularly the liquid, is induced, resulting in a mixed movement / flow of the liquid, which causes mixing. Therefore, by supplying a fluid, a flow is induced, especially within the container, which leads to uniform mixing of the contents. By supplying a fluid, the contents of the container can be mixed flexibly and efficiently, and the mixing can be easily controlled by setting the amount of fluid supplied.

[0018] The mixture can be used in other chemical, biochemical and / or biological processes and / or procedural steps, and the mixture can be transferred to another container, particularly for this purpose. Preferably, the mixture is removed from inside the container using an instrument for removing liquid, such as a pipette or syringe. In this case, the instrument can therefore comprise a pipette or syringe, and this instrument may be designed in particular as a robot and comprise multiple pipettes and / or syringes.

[0019] In the embodiment of the present invention, the components may include, for example, cells (or a plurality of cells), and may consist particularly of cells. Alternatively, the liquid may further include biomolecules, and the liquid may be a suitable solvent, and may particularly include water.

[0020] Alternatively, the components may further include magnetic particles (or a plurality of magnetic particles), and in particular may consist of magnetic particles. In this case, the method according to the present invention may include the magnetic particles being collected by a magnet on the wall of a container. Furthermore, the collected magnetic particles may be resuspended. In particular, these steps can be carried out by moving the magnet. In practice, molecules in the liquid (such as biomolecules) can bind to the magnetic particles. The magnetic particles and molecules bound to the magnetic particles may be transferred into a second container, and / or the magnetic particles and molecules bound to the magnetic particles may be separated from each other, and / or the biomolecules may be concentrated by an elution buffer and a washing step.

[0021] The fluid is preferably a gas, and the gas particularly includes CO2 and / or oxygen and / or an inert gas and / or air. When the fluid is a gas, this has the advantage that the volume of the liquid inside and the concentration of the components do not change by supplying the fluid. In this case, the gas can be introduced in the bottom / lower / downward region of the container in such a manner that the liquid and components are mixed and the components are dispersed in the liquid, and that the liquid flows as the gas rises through the liquid.

[0022] In practice, the supply of the fluid may be controlled by an adjustment element, and the fluid may be supplied from a source (such as a gas container or a gas bottle) to the container via a conducting element (such as a pipe / pipeline).

[0023] The fluid may be in a liquid aggregate state when supplied, and particularly may be a liquid. Thus, in particular, the concentration of the components may be adjusted by supplying the fluid, and mixing and concentration adjustment may be carried out simultaneously. Alternatively, the fluid may be supplied in a liquid aggregate state and then can transition to a gas aggregate state in the liquid.

[0024] In this way, for example, by using liquid nitrogen, a cooling effect can be achieved in addition to mixing.

[0025] According to the present invention, a mixing device for use in the method according to the present invention is further proposed. In this case, the mixing device includes a container having an interior for receiving the contents, and the container includes an opening for receiving and / or removing the contents and a connecting element separated from the opening. The container is replaceable, that is, the container can be removed from the mixing device and replaced with another container. For this purpose, the mixing device includes a housing having a supply element for supplying the fluid into the interior of the container and a receiving element for receiving the container. Further, the container is disposed within the receiving element, and the supply element can be disposed on the connecting element in such a manner that the fluid can be supplied into the interior of the container.

[0026] In fact, the mixing device can include a plurality of supply elements and / or connecting elements (particularly, disposed at different positions of the container / scattered evenly throughout the container) in order to achieve flexible control and increased mixing. In an implementation of the present invention, the supply element can include an adjustment element for setting the flow rate of the fluid, and the adjustment element is a nozzle and / or a valve (such as a duckbill check valve) and / or can include this. The adjustment element as well as other device components and procedure steps can be controlled by a control unit. In this case, the mixing device includes a control unit.

[0027] In a preferred embodiment, the container may be supported on the support surface of the receiving element, and the support element may comprise a fixing element. In this case, the supply element can be made displaceable relative to the receiving element such that the container can be fixed between the fixing element and the support surface after being placed in the receiving element. In this case, the mixing device can comprise a moving device, which can be connected to the supply element or be part of the supply element, and thus the supply element can be moved by the moving device. For this purpose, the moving device can comprise a drive, in particular a motor (such as an electric motor, especially a servo motor, etc.). In this case, the supply element can comprise rails such that the movement of the supply element (and thus also the movement path) is predetermined by the rails. Alternatively, the supply element may be arranged on an element / support / mount that is moved by the drive unit.

[0028] The connection element is preferably a connection opening separated from the opening, and the supply element comprises a supply pipe, which can be introduced / inserted into the connection opening in such a way that fluid can be supplied 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 supply pipe can be inserted. The sealing element can in particular be designed in the form of a membrane (for example, made of polymer / plastic), and the opening is closed by the membrane. In this case, the supply pipe can be inserted / pierced through the membrane in such a way that fluid can be supplied into the interior of the container. The supply pipe is preferably designed as a needle / a piercing-shaped needle. Furthermore, such a sealing element / such a membrane may be provided in the opening. Alternatively, or furthermore, the mixing device can comprise a source connected to the supply element via a conducting element, and the fluid can be conducted from the source to the supply element via the conducting element.

[0029] In practice, the container may be designed as a "consumable", i.e., as a consumable material introduced into the mixing device, and may be disposed of after the corresponding process or procedure has been carried out. Subsequently, a new container can be introduced into the mixing device. Thus, the container is replaceable.

[0030] The container according to the present invention is particularly preferably chemically resistant, pH resistant, and heat resistant. Furthermore, the container may be designed to hold at least 10 ml of liquid / contents, preferably 10 ml to 500 ml or 20 ml to 100 ml, and particularly preferably 20 ml to 50 ml of liquid / contents. In this case, the container may contain, and particularly preferably be made of, a polymer. Examples of polymers are polyethylene, polypropylene, polyethylene terephthalate, and / or polyether ketone.

[0031] The contents of a container may include a liquid and, as components, various magnetic particles / cells / molecules, other particles, or components that settle in the liquid. In this case, the liquid and the settling magnetic particles / cells / molecules, other particles, or components can be mixed by introducing a fluid. Therefore, pre-filled containers are provided in particular and can be dispersed as "consumables." The miscibility ensures that the concentration of the settling magnetic particles / cells / molecules, other particles, or components is the same for each volume of the liquid / contents removed (i.e., a mixture / predeterminable amount) which can then be used in other steps or processes, for example, to process biomolecules. The mixing device / container may, in particular, be for providing a liquid having settling magnetic particles / cells / molecules, other particles, or components.

[0032] In one embodiment, the mixing apparatus may further include a magnet, which may be positioned in the container such that magnetic particles that can be carried inside can be fixed to the inside of the container. This allows the liquid to be discharged and / or new liquid to be supplied without removing the magnetic particles from the inside. In this case, the magnet may be a permanent magnet and / or an electromagnet. When discharging (or supplying) the liquid, an instrument for removing the liquid, such as a pipette, can be used so that the liquid can be removed from the container after the molecules have been immobilized on the surface of the magnetic particles. This can be done through an opening in the container. The instrument for removing the liquid may be a syringe or another suitable instrument.

[0033] In practice, the container and mixing apparatus according to the present invention are preferably used to provide a liquid containing magnetic particles or biomolecules for other processes, such as processing processes (in which case equivalent / reproducible concentrations of magnetic particles may be provided in each volume of the liquid to be removed). Alternatively, the container according to the present invention may be a reaction vessel, and the mixing apparatus may be a bioreactor, in which the biomolecules containing magnetic particles are processed.

[0034] When used as a reaction vessel, the magnet can be positioned in the vessel / mixing apparatus such that magnetic particles can move freely within the vessel during the reaction steps and can be fixed within the vessel during the washing steps by changing the position of the magnet. The magnet can be made movable, in particular, such that the magnet is positioned in the vessel at a first position, fixing the magnetic particles, and then the magnetic particles become movable by moving the magnet to a second position. Naturally, the vessel can also be moved relative to the magnet in an automated apparatus, for example, to achieve the same effect.

[0035] In the method according to the present invention, the contents may be initially introduced into the container, which is of optional choice. The contents may include, for example, magnetic particles and a liquid, which can be mixed inside by supplying the fluid, thereby preventing the sedimentation of the magnetic particles.

[0036] This method can, in particular, provide a liquid having sinking magnetic particles / cells / molecules, other particles or components, in which the sinking magnetic particles / cells / molecules, other particles or components and the liquid (as contents) are introduced into a mixing device, and the liquid and the sinking magnetic particles / cells / molecules, other particles or components are mixed inside. In this case, particularly preferably, a predetermined volume of the liquid / contents can be removed from the container while the fluid is being mixed / supplied.

[0037] Furthermore, this method can be a method for processing biomolecules, in which magnetic particles and a liquid containing biomolecules (as contents) may be introduced into a mixing device, and the liquid and magnetic particles are mixed inside. This method may further include fixing the magnetic particles in the container to the container by placing a magnet in the container, and / or removing the liquid containing the magnetic particles from inside using an instrument for removing the liquid.

[0038] The method according to the present invention, in particular, involves mixing the liquid with the sinking magnetic particles (including magnet particles) / cells / molecules, other particles or components by the action of the fluid on the contents, so that the liquid has a uniform concentration of sinking magnetic particles / cells / molecules, other particles or components.

[0039] Additionally, or alternatively, the fluid supply is designed to keep sinking particles, cells, molecules, or components suspended in such a manner that sedimentation at the bottom of the container is prevented. Furthermore, the mixing or swirling process is improved by allowing particles to remain freely suspended in the container and / or by preventing or reducing foam coagulation. Advantageously, allowing magnetic particles to remain freely suspended and / or avoiding sedimentation of magnetic particles improves biochemical reactions within the mixing apparatus, i.e., the immobilization of biomolecules.

[0040] Within the framework of the present invention, the term biomolecule may be understood to mean DNA, RNA, nucleic acids, proteins, start sequences, monomers, or other bioactive molecules. Such biomolecules may be processed in a mixing apparatus according to the present invention and introduced into a container according to the present invention (preferably in a liquid). The processing may include, among other things, a washing step and / or a reaction step, which may be carried out with magnetic particles. Within the framework of the present invention, the washing step is a procedure step in which the liquid is removed from the container after mixing, and therefore after contaminants have been separated from the magnetic particles, particularly those having sticky biomolecules. Rinsing with a rinsing fluid may also be part of the washing step. Within the framework of the present invention, the reaction step is a procedure step in which biomolecules bound to magnetic particles are transformed, bound to particles, or extended (chain extension, e.g., PCR "polymerase chain reaction"). In particular, contaminants should be understood as substances that have not reacted completely or that have not bound to the magnetic particles, solvent, by-products and contaminants, and mixtures of two or more of the foregoing.

[0041] Within the framework of the present invention, the liquid or contents may be a solution or suspension, in particular a mixture of liquid and magnetic particles or a reaction mixture made of biomolecules and / or reagents and / or contaminants.

[0042] Within the framework of the present invention, “magnetic particles” (also called “magnetic beads”) can be understood as particles in the micrometer or millimeter range. Furthermore, magnetic particles can be porous. Biomolecules can be bonded to the surface of magnetic particles, in particular by thiol groups and / or amino groups and / or hydroxy groups and / or carboxyl groups and / or carbonyl groups and / or ester groups and / or nitrile groups and / or amine groups and / or other functional groups or other interactions. Magnetic particles can also be nickel-coated particles or other ferromagnetic or paramagnetic particles. Magnetic particles typically have a diameter of about 1 micrometer. Within the framework of the present invention, about 1 micrometer should be understood as 0.5 to 1.5 micrometers, particularly 0.7 to 1.3 micrometers, and especially preferably 0.9 to 1.1 micrometers.

[0043] Therefore, the aforementioned magnetic particles can be functionalized magnetic particles that bind biomolecules such as DNA in the sample material. The magnetic particles can then be immobilized using magnetic force and thus separated from the reagent solution. The reagent solution is removed, and then the biomolecules can be separated from the particles. The magnetic particles should preferably be added in precise amounts. This is made possible by the apparatus and method according to the present invention. The mixing apparatus can be used for the preparation of any enzymatic procedure containing nucleic acids or for preparation in any enzymatic procedure (e.g., polymerase chain reaction (PCR), isothermal DNA amplification, reverse transcription). The biomolecules bound to the magnetic particles can be separated from the surface of the magnetic particles and used later.

[0044] The present invention will be described in more detail below with reference to the drawings, with regard to exemplary embodiments. [Brief explanation of the drawing]

[0045] [Figure 1] This is a cross-sectional view of a first exemplary embodiment of the mixing apparatus according to the present invention. [Figure 2A] This is a plan view of a receiving element according to the present invention, which has a container. [Figure 2B] Figure 2A is a side view of the receiving element according to the present invention, which has a container. [Figure 3] This is a schematic diagram of a second exemplary embodiment of the mixing apparatus according to the present invention. [Figure 4] This is a schematic diagram of a mixing apparatus according to the present invention, which includes a pipette. [Modes for carrying out the invention]

[0046] A mixing apparatus is, in particular, an apparatus having a container in which a liquid containing magnetic particles can be provided under configurable, and therefore as optimal as possible, conditions. Therefore, the operation of a mixing apparatus is, in particular, to apply chemical, biochemical, or biotechnological properties that enable or provide free bodies to chemical and biological processes.

[0047] Factors that can be controlled or monitored within a mixing apparatus include the composition of the contents, gas supply, temperature, pH value, and sterility. The mixing apparatus can serve to provide a liquid with a predetermined concentration of sedimentary magnetic particles / cells / molecules / other particles and / or components, or to acquire and process biomolecules. Compound processing and decomposition can also be carried out within the mixing apparatus.

[0048] Figure 1 shows a cross-sectional view of a first exemplary embodiment of the mixing apparatus 11 according to the present invention used in the method according to the present invention.

[0049] The mixing device 11 comprises a housing 4 and a container 1, the container 1 having an interior 17. This interior 17 is suitable for receiving contents such as liquids or components. In this case, the container 1 further comprises an opening 2. Through this opening 2, the contents can be removed from the interior 17 of the container, or another substance can be added to the contents of the interior 17 by pipette. The container 1 may be connected to a supply element 5 via a connecting element 3 different from the opening. Fluid can be introduced into the interior 17 of the container 1 via this supply element 5, and the supply element 5 can be located in or within the housing 4. In this exemplary embodiment, this is done via a side chamber 31, which is connected to the interior 17 via a side opening 32 at the bottom of the container 1.

[0050] Furthermore, the housing 4 is equipped with a receiving element 6, which can receive the container 1. In this case, the container 1 is positioned in the receiving element 6 such that the supply element 5 can be positioned in the connecting element 3 in such a manner that fluid can be introduced into the interior 17 of the container 1 (by introducing the supply pipe 18 into the connecting element 3).

[0051] By introducing this fluid into container 1, the liquid already contained within container 1 begins to move. This mixes the liquid with the components also contained in container 1, thereby achieving a uniform concentration of the components in the liquid.

[0052] The mixing apparatus 11 shown herein also has a source 13 located within or in the housing 4, the source 13 being connected to the supply element 5 by a conduction element 12, so that the fluid can travel from the source 13 through the conduction element 12 to the supply element 5, and from there into the container 1.

[0053] Furthermore, the receiving element 6 has a support surface 8, and the container 1 is supported on the support surface 8 of the receiving element 6. The supply element 5, in this case, has a fixing element 9, and the supply element 5 can be displaced relative to the receiving element 6 by moving the fixing element 9 onto the container 1, and after being positioned within the receiving element 6, it can be fixed between the fixing element 9 and the support surface 8. Since the container 1 is positioned on the spring 20 via the support surface 8 (or via the support element), the container 1 can be tightened between the fixing element 9 and the support surface 8 because the spring 20 can apply pressure to the container 1 positioned within the receiving element 6.

[0054] Figure 2A shows a plan view of the receiving element 6 according to the present invention, which has a container 1.

[0055] The container 1 is positioned within the receiving element 6 such that the receiving element 6 surrounds the container 1. This allows access to the interior 17 of the container 1 from above through the opening 2.

[0056] The fixing element 9 can be pushed onto the wall 19 of the container 1, thereby fixing the container 1 within the receiving element 6. In this case, the fixing element 9 is movably positioned within the housing 4 / receiving element 6 such that the container 1 can be inserted into the receiving element 6 and is fixed within the receiving element 6 by moving the fixing element 9 onto the wall 19.

[0057] Figure 2B shows a side view of a receiving element 6 having a container 1. In this case, the support surface 8 is positioned on the platform 22 by a cylindrical pressure transmission element 80, and the platform 22 is positioned by a spring 20 such that pressure can be applied to the container 1 placed inside the receiving element 6 via the platform 22 and the support surface 8 by the spring 20.

[0058] Figure 3 shows a schematic diagram of a second exemplary embodiment of the mixing apparatus 11 according to the present invention.

[0059] The interior 17 of container 1 contains liquid 14 and component 15 placed inside container 1. Container 1 has an opening 2 for receiving and / or removing the contents 14, 15, and a supply element 5 is located on the underside of container 1, so that gas can be supplied into the interior 17 of container 1 via the supply element 6 through the connecting element 3.

[0060] The gas is supplied to the liquid 14 inside the interior 17 via the supply element 5 along direction A, and the liquid 14 starts moving in such a manner that the liquid 14 and component 15 are mixed by the addition of the gas, and as a result, component 15 is dispersed in the liquid 14.

[0061] The gas is introduced at the bottom of the container 1 in such a manner that the liquid 14 and component 15 are mixed, and that the gas causes the liquid 14 to flow as it rises through the liquid 14.

[0062] A predetermined amount of the contents 14, 15 can be removed from the interior 17 through the opening 2 while supplying gas.

[0063] Figure 4 shows a schematic diagram of a portion of the mixing apparatus 11 according to the present invention, which has a pipette 16, and the supply element is not explicitly shown.

[0064] In this example, the pipette 16 can be passed through the opening 2 of the container 1 to the interior 17 and immersed in the liquid 14 containing the component 15 placed inside the container 1. By manipulating the pipette 16, the liquid 14 and component 15 are kept suspended by the fluid flow in the liquid and can be collected and removed from the container 1 through the opening 2. Similarly, the liquid 14 and / or component 15 can be supplied to the interior 17 of the container 1 through this opening 2.

[0065] In this exemplary embodiment, component 15 is a plurality of magnetic particles 15. The mixing device 11 is equipped with a magnet 7, which is positioned in the container 1 in such a manner that the magnetic particles 15 present in the container 1 are fixed in place. This allows the liquid 14 to be introduced into the interior 17 of the container 1 via the opening 2 by the pipette 16 and then discharged by the pipette 16 without the magnetic particles 15. When the magnet 7 is moved away from the container 1, the liquid 14 can be removed with a predetermined concentration of magnetic particles 15.

[0066] Although the present invention is depicted and described in detail in the figures and the foregoing description, such depictions and descriptions should be considered illustrative or exemplary and not limiting.

[0067] The present invention is not limited to the disclosed embodiments. Other variations of the disclosed embodiments can be understood and achieved by a skilled practitioner when practicing the claimed invention in consideration of the figures, this 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 the existence of multiple elements or steps. The mere fact that certain means are repeated in different dependent claims does not mean that such means cannot be used advantageously in combination. Reference numerals in the claims should not be construed as limiting the scope.

Claims

1. A method for providing a mixture from the inside (17) of a container (1), wherein the contents (14, 15) inside (17) comprise a liquid (14) and a component (15), the container (1) comprises at least one opening (2) for receiving and / or removing the contents (14, 15), and a supply element (5) is positioned in the container (1) such that a fluid can be supplied into the inside (17) of the container (1) via the supply element (5). a) A step of supplying the fluid to the liquid (14) inside (17) via the supply element (5), wherein the liquid (14) begins to move in such a manner that, upon addition of the fluid, the liquid (14) and the component (15) are mixed, and as a result, the component (15) is dispersed in the liquid (14). b) Providing the mixture by removing a predetermined amount of the contents (14, 15) from the interior (17) while supplying the fluid; Methods that include...

2. The method according to claim 1, wherein the component (15) includes cells, and more particularly consists of cells.

3. The method according to claim 1 or 2, wherein the liquid (14) contains biomolecules.

4. The method according to any one of claims 1 to 3, wherein the component (15) includes magnetic particles (15), and 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) located on the wall of the container (1).

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

7. The method according to any one of claims 4 to 6, wherein the molecules in the liquid (14) bond to the magnetic particles (15).

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

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

10. The method according to claim 9, wherein the biomolecule is concentrated by an elution buffer and a washing step.

11. The method according to any one of claims 1 to 10, wherein the mixture is transferred to another container.

12. The method according to any one of claims 1 to 11, wherein the mixture is removed from the inside (17) of the container (1) by a pipette (16) or syringe.

13. The aforementioned fluid is a gas, particularly CO ２ The method according to any one of claims 1 to 12, wherein the fluid is and / or oxygen, and / or the fluid contains an inert gas.

14. The method according to claim 13, wherein the gas is introduced at the bottom and / or underside of the container (1) in such a manner that the liquid (14) and the component (15) are mixed and the component (15) is dispersed in the liquid (14), and the gas causes the liquid (14) to flow as it rises through the liquid (14).

15. The method according to any one of claims 1 to 14, wherein the supply of the fluid is controlled by a control element.

16. The method according to any one of claims 1 to 15, wherein the fluid is supplied from a source (13) to the container (1) via a conductive element (12).

17. The method according to any one of claims 1 to 16, wherein the fluid is in a liquid aggregate state when supplied, and in particular is the liquid (14).

18. A mixing apparatus used in the method according to any one of claims 1 to 17, A replaceable container (1) having an interior (17) for receiving contents (14, 15), comprising an opening (2) for receiving and / or removing the contents (14, 15) and a connecting element (3) separated from the opening (2), A housing (4) having a supply element (5) for supplying fluid into the interior (17) of the container (1) and a receiving element (6) for receiving the container (1) Equipped with, A mixing device in which the container (1) is placed inside the receiving element (6), and the supply element (5) is placed on the connecting element (3) in such a manner that the fluid can be supplied into the interior (17) of the container (1).

19. The mixing apparatus according to claim 18, wherein the supply element (5) includes an adjustment element for setting the flow rate of the fluid.

20. The mixing apparatus according to claim 18 or 19, wherein the container (1) is supported on the support surface (8) of the receiving element (6), the supply element (5) comprises a fixing element (9), and the supply element (5) can be displaced 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 it has been placed inside the receiving element (6).

21. The mixing apparatus according to any one of claims 18 to 20, wherein the connecting element (3) is a connecting opening (3) separated from the opening (2), and the supply element (5) comprises a supply pipe (18), the supply pipe (18) can be introduced into the connecting opening (3) in such a manner that the fluid can be supplied into the interior (17) of the container (1).

22. The mixing apparatus according to claim 21, wherein the connecting opening (3) is closed by a membrane, and the supply pipe (18) can be inserted through the membrane in such a manner that the fluid can be supplied into the interior (17) of the container (1).

23. A mixing apparatus according to any one of claims 18 to 22, comprising a source (13) connected to the supply element (5) via a conduction element (12), wherein the fluid can be conducted from the source (13) to the supply element (5) via the conduction element (12).

24. The mixing apparatus according to claim 19, wherein the adjustment element comprises a nozzle and / or a valve.

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