EP2928605B1 - Reaction vessel having a magnetic closure - Google Patents
Reaction vessel having a magnetic closure Download PDFInfo
- Publication number
- EP2928605B1 EP2928605B1 EP13805819.3A EP13805819A EP2928605B1 EP 2928605 B1 EP2928605 B1 EP 2928605B1 EP 13805819 A EP13805819 A EP 13805819A EP 2928605 B1 EP2928605 B1 EP 2928605B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- reaction vessel
- subdivision
- state
- magnetic field
- movable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Not-in-force
Links
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5082—Test tubes per se
- B01L3/50825—Closing or opening means, corks, bungs
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/041—Connecting closures to device or container
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/046—Function or devices integrated in the closure
- B01L2300/047—Additional chamber, reservoir
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/046—Function or devices integrated in the closure
- B01L2300/049—Valves integrated in closure
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
- B01L2400/0633—Valves, specific forms thereof with moving parts
Definitions
- the invention relates to a reaction vessel having a first and a second subdivision, wherein in a first state of the reaction vessel, the first is separated from the second subdivision and in a second state, the first is connected to the second subdivision. Furthermore, the use of the reaction vessel in a method for the detection of proteins and / or nucleic acids and in a method for the amplification of nucleic acids is part of the invention. The invention further includes a method for converting the one into the other of the two states of the reaction vessel.
- the patent application WO 00/12675 describes an apparatus having a first hollow elongate cylinder with a closed end and therein a plurality of chambers, a second hollow elongated cylinder internally positioned contiguously adjacent to the first cylinder and capable of relative rotation.
- a closed position A a sample is introduced into the second cylinder.
- the second cylinder contains reagents for extraction of nucleic acids.
- the second cylinder is rotated to an open position B.
- the extracted nucleic acid remains in a solid state in an upper chamber as the fluid flows into a reservoir.
- the second cylinder is rotated to the closed position A so that the second cylinder is sealed, water is added, and a cover is closed.
- a reaction bead from one chamber is added to the second cylinder.
- the reaction bead contains those for amplification necessary enzymes that are resuspended in the water.
- the amplification takes place on the solid phase, which contains the extracted nucleic acids.
- the second cylinder is rotated relative to the first cylinder to a washout position C.
- the reaction mixture of the amplification may enter the detection chamber and is absorbed by a pad.
- the reaction mixture is absorbed by a strip.
- colored microparticles bind to the haptens from the amplification reaction and move to the capture zone on a membrane where they form a visible detection line if the target sequence is present.
- the patent application WO 2006/050636 A1 describes a PCR reaction tube having a tubular body and a cap, wherein the inner surface of the cap is provided with an inner tube and the inner tube is provided with at least one hole.
- the reaction vessel is suitable for performing nested PCR, whereby reagents for the nested inner part of the PCR in the inner tube can be provided. After the first part of the PCR is complete, the reaction vessel can be centrifuged or exposed to vibration so that the reagents are released from the inner tube. Thereafter, the second part of the PCR can be performed.
- the U.S. Patent 5,576,197 discloses a reaction vessel for PCR in which wax is applied to the inner surface.
- the wax is at a position that is approximately at the height of the meniscus of a filled PCR mix.
- the wax does not completely close the volume of the vessel.
- the wax is solid at temperatures below 40 ° C and liquid at PCR temperatures. When the wax liquefies, it spontaneously covers the surface of the PCR mix.
- the wax is present in an amount sufficient so that when the wax is liquefied, it completely covers the surface of the PCR mix.
- the molten fat or wax creates a vapor barrier to minimize evaporation of the solution during the thermal PCR cycles and, upon cooling, after the amplification is complete, again forms a solid barrier that, inter alia, reduces the likelihood of contamination of the following reactions.
- the patent application WO 2008/047272 A2 discloses an apparatus suitable for use in the amplification and detection of nucleic acids.
- the device comprises a simple sample tube having a chamber which is preferably open at the top so that its contents flow out when the sample tube is turned upside down.
- the sample tube further comprises a lid containing a microarray for nucleic acids to which amplified samples can hybridize DNA.
- a bio-valve thin-film device which comprises a plurality of chambers, for example a preparation chamber, a buffer chamber, a reaction chamber and a waste chamber. Between the chambers arranged magnetic valves can be opened and closed by the force of a movable permanent magnet.
- the patent US 3,424,820 discloses a method in which two substances are mixed together by the action of a magnetic field. In a first state, these substances are separated by a liquid having a certain density. A perforator can be moved by means of magnetic fields in such a way that the separating liquid perforated and as a result, the two substances are mixed together.
- the patent application US 2006 0116270 A1 discloses a system for maintaining a liquid separation comprising a liquid container, a solenoid-operated valve within the liquid container, and a centrifuge that positions the valve so that liquid can flow past the valve, while the centrifuge is running.
- the valve contains an iron material that is placed in a plug.
- the plug interacts with a solenoid of a valve actuator. This interaction causes the valve to be selectively opened and closed. When the valve is open, the flow of a sample around the valve is possible. When the valve is closed, the flow of the sample around the valve is blocked.
- the invention has for its object to provide an improved reaction vessel and a use for such an improved reaction vessel.
- an object of the invention is to provide a method using the improved reaction vessel available.
- the object is achieved by a reaction vessel with the features of claims 1, 2 and 3.
- the task is also solved by use of the reaction vessel in a process for purification, modification and / or detection of proteins and / or nucleic acids.
- a further solution of the object according to the invention consists in a method for transferring one into the other of the two states of the reaction vessel.
- the reaction vessel is a vessel capable of reacting in it.
- the reaction vessel is closable, z. B. with a lid.
- the preferred closed reaction vessel is liquid-tight with respect to its environment, particularly preferably it is gas-tight.
- the reaction vessel can in principle assume any shape and size.
- the reaction vessel is substantially cylindrical, circular cylindrical or conical, more preferably, the reaction vessel has both a substantially circular cylindrical portion and a substantially conical portion.
- the reaction vessel z.T. plane walls, in particular e.g. a rectangular base, as in commercial cuvettes.
- the volume of the reaction vessel is less than 500 ml, more preferably less than 100 ml, more preferably less than 50 ml, more preferably less than 25 ml, more preferably less than 10 ml, most preferably less than 5 ml, most preferably less than 2 , 5 ml, particularly preferably less than 1 ml, more preferably less than 0.5 ml and most preferably less than 0.25 ml.
- the preferred volume of the reaction vessel is greater than 0.05 ml, more preferably greater than 0 , 1 ml.
- the reaction vessel is at least partially, particularly preferably in its entirety made of one or more inert materials such as glass or plastic, preferably polyethylene, polypropylene or silicone.
- the reaction vessel is preferably transparent, so that an optical inspection of the opening mechanism or the opening success is possible.
- the reaction vessel has at least two subdivisions.
- the first In the first state, the first is separated from the second subdivision, so that no liquid - preferably also no gases - can flow from one of the two subdivisions into the other.
- the first In the second state, the first is connected to the second subdivision and liquids - preferably also gases - can flow from one of the two subdivisions into the other.
- One subdivision may adjoin the other, one may comprise the other.
- the partitions may also be spaced apart so that they do not abut each other.
- the sealed reaction vessel encloses the subdivisions.
- the subdivisions are fixed in the reaction vessel.
- the subdivisions are connected. This can be done reversibly, so that the reaction vessel from the second can be converted back into the first state.
- the second state is irreversibly produced, for. When a wall separating the first from the second partition is destroyed in the state change.
- the reaction vessel may preferably have further subdivisions.
- the further subdivisions may be in the first and / or second state with the first and / or the second subdivision and / or interconnected or separated from each other.
- the reaction vessel may preferably assume further states.
- one or more of the further subdivisions may preferably be connected to or separate from one or more of the further subdivisions and / or with the first and / or the second subdivision.
- the first and / or second state can be converted into the other of the two states by the action of one magnetic field.
- the first can be separated from the second subdivision or the first can be connected to the second subdivision.
- the reaction vessel can be used in a process for the purification, modification and / or detection of proteins and / or nucleic acids.
- a first reaction can take place in the first subdivision, which is initially separated from the second subdivision.
- the first reaction may, for. Example, the proliferation of nucleic acids by PCR or the purification of nucleic acids or proteins from a sample.
- the second reaction may, for. B. be a detection reaction to a specific nucleic acid or a specific protein. In this way, nucleic acids and proteins can be analyzed in a particularly simple and low-contamination manner.
- the modification may be, for example, in a digestion of a sample or label of a protein or a nucleic acid.
- the reaction vessel can be used according to the invention in a method for the amplification of nucleic acids.
- a part of the reagents required for the duplication are present in the first subdivision.
- the remaining part is preferably contained exclusively in the second subdivision.
- the first partition may be connected to the second partition by the action of a magnetic field. Due to the separate storage of the two parts a higher shelf life of the reagents can be achieved.
- the particular desired reaction conditions can be optimally adjusted by the targeted presence or absence and the concentration of the respective reagents in each of the two parts.
- the invention further includes a method for changing the state of the reaction vessel.
- the reaction vessel is exposed to a magnetic field.
- a magnetic field In this case, either no magnetic field is present before exposure or a previously existing magnetic field is changed when exposed to alignment and / or field strength. This can be accomplished by moving the reaction vessel relative to an existing magnetic field.
- a non-constant magnetic field may be used, e.g.
- a magnetic field can be generated or changed by means of an electromagnet.
- the invention it is advantageously possible to store two different liquids in two subdivisions of a reaction vessel without the liquids mixing.
- one partition can be connected to the other so that the two liquids can mix with one another.
- the abandonment of the opening of the reaction vessel for mixing the liquids can be the automation of Simplify reactions.
- waxes are used to separate two liquids in a reaction vessel, which are heated to mix the liquids; as soon as the wax cools down and hardens, mixing is no longer possible or greatly impeded.
- an inert material can be used to separate the liquids in the subdivisions, no mixing is necessary for mixing the liquids, whereby an impairment of reactions can be avoided.
- the invention makes it possible to produce a structurally simple, inexpensive reaction vessel.
- the first and second subdivisions are separated by a movable shutter.
- the movable closure may in principle take any shape, but preferably the movable closure is spherical or cylindrical in shape.
- the connection of the first with the second subdivision can be made reversible.
- a separation of initially associated subdivisions can be brought about by a movement of the movable closure.
- Movement of the movable shutter which leads to the change of state of the reaction vessel, may consist of a translatory or rotational movement or a combination of the two types of movement.
- the movable shutter may consist of a single or multiple elements.
- the movable shutter is a ball that closes off the first partition from the second partition.
- a one-piece movable closure can advantageously be produced particularly easily.
- the movable closure consists of a flap and a latch.
- the flap can separate the two divisions against each other, the latch can lock the flap.
- the latch In order to bring about the state change, in this embodiment, first of all, the latch must be moved, so that the flap is released. Only then can the flap be moved. With such a Two-piece movable closure can be prevented that the change of state, for example, during an optional centrifugation or shocks generated accidentally, for example, by an un-padded case on the ground.
- Such an unintentional, premature change of state can preferably be prevented by the bar according to the invention also in cases in which a high pressure builds up in the closed subdivision (be it by a high temperature, by the use of very volatile chemicals, by reaction-induced gas formation or by low external pressure eg during transport).
- the movable shutter includes a magnetic component.
- the movable closure consists of the magnetic component.
- the preferred magnetic component is cylindrical or spherical.
- the magnetic component of the movable shutter can be moved. In an inhomogeneous magnetic field, the magnetic component may move from a weaker location to a stronger magnetic field location.
- the magnetic component of the movable shutter can be aligned by the action of the magnetic field along the field line of the magnetic field. This can be done both in an inhomogeneous as well as in a homogeneous magnetic field. Due to the magnetic component of the movable shutter, the state change can be achieved particularly easily by the action of the magnetic field.
- the movable shutter may e.g. consist of a magnetic component (e.g., a ferromagnetic material) provided with a plastic coating for better sealing and / or corrosion protection.
- the movable shutter may be e.g. Also, consist of a non-magnetic body, which is provided with a ferromagnetic layer, which represents the magnetic component.
- the movable shutter with the magnetic component may be incorporated in a partition wall between the first and second divisions.
- the magnetic component By acting on the magnetic field, the magnetic component can move and / or align so as to form a connection between the first and second subdivisions.
- parts or the entire partition wall between the first and second Subdivision of the magnetic component Preferably, parts or the entire partition wall between the first and second Subdivision of the magnetic component. In this way, the reaction vessel can be produced with a particularly low cost of materials.
- the magnetic component may include a ferromagnetic material or be made of a ferromagnetic material.
- the magnetic component contains iron. By using iron in the magnetic component, the manufacturing cost of the reaction vessel can be kept particularly low.
- the magnetic component is at least partially permanent magnetic.
- the magnetic component consists entirely of a permanent magnet.
- an applied magnetic field it is possible for an applied magnetic field to exert a greater force on the magnetic component than if the magnetic component consists solely of a ferromagnetic, non-magnetized material.
- a permanent magnet that aligns the magnetic component in a magnetic field.
- the magnetic component may align in a magnetic field along the field lines.
- the alignment along the field lines produces a rotational movement of the magnetic component, by means of which the first and the second subdivision can be connected to one another or separated from one another.
- the movable closure in the first or second state is non-positively connected to the reaction vessel.
- an area of the movable shutter on a surface of the reaction vessel is such that a force along the surface normal generates sufficient friction between the surfaces to hold the movable shutter in the reaction vessel.
- exposure to the magnetic field may exert sufficient force on the magnetic component of the movable shutter to overcome the friction of the two surfaces, such that the movable shutter moves in the reaction vessel and the first and second subdivisions are connected or disconnected can.
- the movable closure and the adjacent parts of the reaction vessel can take any shape.
- the movable closure of a magnetic rotationally symmetrical body preferably of a permanent magnetic circular cylinder or a permanent magnetic ball.
- the diameter of the body is preferably between 0.1 and 10 mm, more preferably between 1 and 6 mm and most preferably between 2 and 4 mm.
- the body can be inserted into a flexible tube which is closed at one end.
- the tube is made of silicone.
- the ratio of the diameter of the body to the inner diameter of the tube is preferably 1: 1, particularly preferably 1.01: 1, particularly preferably 1.03: 1, particularly preferably 1.05: 1, more preferably 1.1: 1, more preferably 1.2: 1, most preferably 1.3: 1, and most preferably 1.5: 1.
- the hose is stretched during insertion of the body so that a tight connection between the hose and the body is formed.
- the tube preferably has a wall thickness between 0.1 and 5 mm, more preferably between 0.5 and 3 mm, and most preferably between 0.7 and 1.5 mm.
- the first subdivision in which a liquid can be located.
- the stiction holding the body in the tube can be overcome so that the body is removed from the tube and the first partition is connected to a second partition comprising the tube.
- the movable closure in the first or second state is positively connected to the reaction vessel.
- a first portion of the movable shutter is arranged to obstruct a second portion of the movable shutter the path that must be traveled to connect the first subdivision with the second subdivision.
- the first portion is a bar which blocks the second portion so that no connection between the first and the second partition can occur.
- the second portion is a hinged lid and the first portion is a bar containing the magnetic component.
- the opening of the lid can by a suitable elastic element, such as a steel spring, or be relieved at a corresponding material of the lid by biasing the lid.
- a suitable elastic element such as a steel spring
- the positive connection of the movable closure allows a particularly secure separation of the first from the second subdivision in the first state. An accidental release of the movable closure, for example in an optional centrifugation step, can be avoided.
- the movable closure in the first or second state is firmly bonded to the reaction vessel.
- the movable closure is irreversibly joined to the rest of the reaction vessel.
- a state change in which the first and second subdivisions are connected together can only be achieved by destroying parts of the movable shutter.
- the movable closure z. B. consist of a film with which the first subdivision is closed.
- a magnetic component present in the film can, by the action of the magnetic field, transmit a force to the film sufficient to destroy the film and thereby connect the first to the second subdivision.
- the movable closure - can be made in one piece with the rest of the reaction vessel except for the magnetic component, preferably predetermined breaking points are attached to the movable closure, so that by the action of the magnetic field, the magnetic component on the movable closure exerts a force to destroy the predetermined breaking points and thus the state change of the reaction vessel leads.
- At least one of the subdivisions is at least partially bounded by a cover of the reaction vessel. With the lid open, the reaction liquid can be filled into at least one of the subdivisions. After closing the lid both subdivisions are preferably closed on all sides. In this embodiment, the filling of liquids is particularly simple.
- the reaction vessel is manufactured in one piece with at least one of the two subdivisions.
- both subdivisions are made in one piece with the reaction vessel.
- the moving one Closure may preferably be made in one piece with the rest of the reaction vessel. In this type of production, the reaction vessel can be produced particularly cheap.
- At least one of the two subdivisions is located in a hollow body which is movable in the reaction vessel.
- this hollow body is not attached in the reaction vessel, but is located loosely in the reaction vessel.
- Such a subdivision may, for. B. are designed as a hose which is closed on one side by the movable closure and on the other side by a solid wall and is not attached to the rest of the reaction vessel.
- an already closed subdivision is introduced into the reaction vessel, which may be advantageous when using particularly volatile reagents.
- the magnetic component may thereby move in the inhomogeneous magnetic field of zones of low magnetic field strength to higher magnetic field strength, wherein preferably the first subdivision is connected to the second subdivision or these two are separated from each other.
- these can be aligned in the direction of the field lines in the inhomogeneous magnetic field and the state change can thus be achieved.
- the state change of the reaction vessel is brought about by a homogeneous magnetic field.
- the magnetic component is preferably permanently magnetic, so that it rotates in the magnetic field and aligns along the field lines.
- the magnetic component can be mounted so that the movable shutter is opened by its rotation.
- the movable shutter may consist of a magnetized ball having a bore.
- the ball may be rotatably mounted in a bearing, so that in an orientation of the ball, the bore is sealed by the bearing.
- the orientation of the permanent magnetic sphere may change so that the bore is no longer obscured and the first is connected to the second subdivision via the bore of the sphere.
- connection of the subdivisions can be made reversible in this way.
- the state change of the reaction vessel is done using a permanent magnet or an electromagnet.
- a permanent magnet By using a permanent magnet, the state change of the reaction vessel can be brought about without an electric device. This is particularly advantageous for mobile use of the reaction vessels.
- An electromagnet can generate the magnetic field necessary for the change of state in a relatively small size. Since the magnetic field can be switched on and off as desired by the electromagnet, no relative movement of the reaction vessel to the electromagnet is necessary to trigger the change of state. Rather, the reaction vessel can be placed firmly in place and the magnetic field switched on at the appropriate time.
- the state change of the reaction vessel using two permanent and / or electromagnets which are arranged such that between the permanent and / or electromagnets, a gap is formed.
- the inventors have found through simulations that such an array of magnets can create an inhomogeneous field whose spatial variation in magnetic flux density is sufficient to provide the necessary force for the change in state of the reaction vessel to the magnetic component.
- the magnitude of the magnetic field gradient is preferably between 0.001 and 10 T / mm, more preferably between 0.01 and 1 T / mm and most preferably between 0.03 and 0.3 T / mm.
- the substances presented in the subdivisions may preferably mix or react with one another.
- the substances preferably come into contact after the connection has been established by gravity.
- it may be necessary to shake the reaction vessel, vibrate or suspend in a suitable form of centrifugation so that the liquid flows out.
- FIG. 1 shows a reaction vessel 1, which is made in this embodiment of polypropylene, polyethylene, silicone or other plastic.
- the reaction vessel 1 consists of an upwardly open circular cylindrical and a lower conical part which is rounded towards the lower end.
- the outer opening 2 on the circular cylindrical part of the reaction vessel 1 can be closed by a cover 3.
- a hose 4 is fixed for example by clamping, gluing or welding.
- the tube 4 is made of silicone and clamped in the lid 3.
- the tube 4 can also be made of other materials, preferably plastics.
- the lid 3 is produced in one piece together with the tube 4 made of polypropylene.
- a liquid or a solid, such as a powder can be filled.
- the tube 4 can be closed by a movable cylindrical closure 5 in this case.
- the tube 4 thus encloses, together with the lid 3 and the movable closure 5, the first partition 6, which is surrounded by the remaining reaction vessel 1 as the second partition 7.
- the circular cylindrical movable shutter 5 includes a magnetic component 8 made of a ferromagnetic or ferrimagnetic material such as iron or ferrite.
- the entire movable closure 5 consists of the magnetic component 8.
- the movable closure 5 can also be provided with a plastic coating or consist of a plastic in which magnetic particles are introduced.
- FIG. 1 In the partial illustration on the top right, a schematic external permanent magnet 9 is brought into the vicinity of the reaction vessel 1 and thus generates an inhomogeneous magnetic field in the reaction vessel.
- the movable shutter 5 moves in the inhomogeneous field in the direction of higher magnetic field strength to the permanent magnet 9 and is removed from the tube 4.
- the liquid previously contained in the tube 4 flows downwards out of the tube 4 out and falls on the liquid which is in the second subdivision 7.
- a centrifugal acceleration of preferably less than 10,000 g (g 9.81 m / s 2 ), particularly preferably less than 1000 g, particularly preferably less than 100 g and very particularly less than 10 g, is usually required for this. It may be sufficient to pick up the reaction vessel and make a suitable circular rapid movement of the hand.
- the reaction vessel 1 can now be moved.
- the movable shutter 5 can also be set in motion by an alternating magnetic field ( FIG. 1 , bottom left) so as to mix the liquids, such as is usually done with a magnetic fish and a magnetic stirrer. Subsequently, by a lateral placement of the external permanent magnet 9, the movable shutter 5 can be removed from the liquid mixture in order to perform an absorption measurement of the liquid with a laser 10 or another light source, eg an LED ( FIG. 1 , below, in the middle and on the right).
- a laser 10 or another light source eg an LED
- reaction vessel 1 is particularly suitable for the detection of nucleic acids using nanoparticles.
- a sample to be tested can be filled into the lower part of the reaction vessel 1 together with PCR mix including primers and polymerase.
- the nucleic acids required for the detection reaction, conjugated with nucleic acids nanoparticles can be filled as a suspension in the tube 4 on the lid 3.
- the lid 3 is now closed with the movable closure 5 and placed on the remaining reaction vessel 1.
- the subdivisions 6, 7 are initially separated from each other.
- the reaction vessel 1 passes through the temperature cycles necessary for carrying out a PCR in a thermocycler. Thereafter, the amplified nucleic acid is present in the liquid at the bottom of the reaction vessel 1.
- the movable shutter 5 is removed from the tube 4 by the action of an inhomogeneous magnetic field, so that the nanoparticle suspension is released and flows to the bottom of the reaction vessel (optionally by exposing the reaction vessel to suitable accelerations, such as by shaking).
- a low-frequency alternating magnetic field is applied, whereby the movable shutter 5 mixes the liquids at the bottom of the reaction vessel 1.
- the low-frequency alternating field can preferably also be generated by movement of the reaction vessel in the vicinity of a (permanent) magnet.
- the nanoparticles can specifically bind to the nucleic acids amplified by PCR via nucleic acids conjugated to them, whereby a change in the absorption spectrum of the liquid can be produced.
- the movable shutter 5 is removed from the liquid by the application of an inhomogeneous magnetic field and held in the upper region of the reaction vessel 1. Subsequently, the concentration of the desired nucleic acids by absorption measurement by means of a laser 10 is determined.
- FIG. 2 schematically illustrates a method with which in.
- Reaction vessel 1 shown a change in state can be brought about.
- a gap 12 is located between two external permanent magnets 9, which in this case each consist of three individual, joined individual magnets 11.
- a reaction vessel 1 with a separate first 6 and second subdivision 7 is moved from left to right through the gap 12 between the two external permanent magnet 9 moves.
- the field strength of the magnetic field in the center of the gap 12 is much stronger than at its edge, so that the movable shutter 5 is pulled out of the tube 4 due to the magnetic component 8 contained therein and the partitions 6, 7 are connected.
- the liquid contained in the tube 4 can thus mix with the liquid present at the bottom of the reaction vessel 1.
- the state change can be brought about here without the use of electricity.
- FIG. 3 shows a reaction vessel 1 in a further embodiment.
- the first subdivision is provided by a tube 4 made of silicone, which is bounded by the lid 3 of the reaction vessel 1 on one side and by the movable shutter 5 on the other side.
- the movable shutter 5 is not designed here as a circular cylinder 13 but as a ball 14.
- the ball 14 may for example consist of a ferromagnetic material or contain a ferromagnetic material. For better sealing and / or for better corrosion protection, the ball 14 may be coated with a plastic material (eg PTFE).
- the diameter of the ball 14 is slightly larger than the inner diameter of the tube 4, so that it is expanded by the ball 14 and thus the sealing effect between the tube 4 and movable closure 5 is improved. Due to the symmetry properties of the ball 14, the movable shutter 5 can be particularly easily inserted into the tube 4 in this embodiment, since it does not have to pay attention to whether the axis of rotation of the movable shutter 5 coincides with the axis of the tube 4. To change the state, an inhomogeneous magnetic field is also applied here, which has a higher field strength, for example, at the bottom of the reaction vessel 1, so that the ball 14 is pulled out of the tube 4 and the subdivisions 6, 7 are connected.
- the magnetic field is selected such that the strongest gradient of the magnetic field strength is at the location of the sphere.
- the liquid flows from the tube 4 to the bottom of the reaction vessel 1.
- an alternating magnetic field eg, by the relative movement of the reaction vessel to a permanent magnet
- the ball 14 mixes the two liquids.
- the ball 14 need not be removed from the liquid, since it is small enough that the light of the laser 10 above the ball 14 can radiate through the liquid.
- FIG. 4 shows an alternative embodiment of the reaction vessel 1 with a fixed to the cover 3 first subdivision 6.
- a recess 15 in which the movable closure is introduced in the form of a ball 14.
- the ball 14 By the action of a magnetic field, the ball 14 can be pulled down from the first partition 6 and then stored laterally on the reaction vessel 1. The liquid contained in the first subdivision 6 can thus flow into the remaining reaction vessel 1 and mix with the liquid at the bottom of the reaction vessel 1.
- the magnetic field can also be designed so that the ball 14 is drawn into the first subdivision 6 and the state change of the reaction vessel 1 is thereby brought about.
- the first partition 6 is not attached to the lid 3, but at an upper wall portion 18 of the reaction vessel 1.
- the reaction vessel 1 is designed so that, when the lid 3 is open, advantageously liquids can be filled both directly into the first subdivision 6 and the second subdivision 7.
- the first partition 6 is closed at its lower end with a spherical movable shutter 5. Again, the inhomogeneous magnetic field can be generated so that in the state change of the movable shutter 5 is pulled into the first 6 or in the second division 7.
- a reaction vessel 1 is divided into an upper, first subdivision 6 and a lower, second subdivision 7 by a partition wall 19 which runs parallel to the plane of the cover 3.
- the movable shutter 5 is in the form of a ball 14. Liquid can be filled only in the second partition 7 when the ball 14 is removed.
- the first subdivision 6 should be empty, in order to avoid that liquid from the first subdivision 6 already gets into the second subdivision 7 during the filling of the liquids.
- FIG. 7 shows a further embodiment of a reaction vessel 1 with a vertical partition wall 19 between the first 6 and second partition 7.
- the partition 19 is located in a plane of symmetry of the reaction vessel 1.
- a spherical, movable shutter 5 can be pulled by the action of a magnetic field in the first 6 or second partition 7, so that the two partitions 6, 7 are connected and at sufficient filling level, the liquids in the partitions 6, 7 are mixed together.
- FIG. 8 the reaction vessel 1 according to the invention is shown, which has a first subdivision 6, which is surrounded by the outer walls of the reaction vessel 1 including the lid 3.
- a second, freely movable subdivision 7 can be added in the first subdivision 6, a second, freely movable subdivision 7 can be added.
- the second partition 7 is formed by a movable hollow body 20, which is permanently closed at one end and in which a movable closure 5 can be sealingly introduced at the other end. If the subdivisions 6, 7 are to be connected to one another, an inhomogeneous magnetic field can be applied, so that the movable shutter 5 with the second subdivision 7 rests against the wall of the reaction vessel 1.
- spacers 21 are mounted, which can keep the opening of the second partition 7 at least as far away from the inner wall of the first partition 6 when acting on the inhomogeneous magnetic field, that there is sufficient space to pull out the movable shutter 5.
- FIG. 9 shows a further embodiment of the reaction vessel 1 according to the invention with an outer first subdivision 6 and a second freely movable therein Division 7 in a movable hollow body 20.
- the second partition 7 is cylindrical and with a spherical, movable closure 5 closed.
- the pulling out of the movable shutter 5 upon the action of an inhomogeneous alternating magnetic field is based here on the lower inertia of the movable shutter 5 in comparison to the remaining second subdivision 7
- the movable shutter 5 has a substantially lower mass than the second subdivision including the liquid 7 contained therein.
- reaction vessel 1 has a lower conical portion 22, in which after filling a liquid, a movable closure 5 - can be pressed - in the form of a ball 14 - here.
- a movable closure 5 - can be pressed - in the form of a ball 14 - here.
- the ball 14 is pulled upwards, so that the subdivisions 6, 7 connect.
- an elastic ring 23 is introduced, which preferably consists of silicone.
- the outer diameter of the ring 23 corresponds to the inner diameter of the reaction vessel 1 and the inner diameter of the ring 23 corresponds to the outer diameter of a circular cylinder 13 (or is slightly smaller), which constitutes the movable shutter 5.
- the movable shutter 5 is pressed into the ring 23.
- the ring 23 is pressed with the movable shutter 5 in the reaction vessel 1, whereby two subdivisions 6, 7 arise.
- the movable shutter can be removed from the ring 23, so that the partitions 6, 7 are connected to each other.
- FIG. 12 is a first subdivision 6 on the lid 7 of the reaction vessel 1 by a film 24 created.
- a magnetic component 8 which in this embodiment is realized as a ball 14.
- a liquid is additionally included in the first subdivision 6.
- the movable shutter 5 in this case consists of two components: the magnetic component 8 (the ball 14) and the film 24.
- the reaction vessel 1 is in an inhomogeneous field, the ball 14 can be pulled down so that the film 24 breaks and a connection between the two divisions 6, 7 arises.
- the connection of the two subdivisions 6, 7 is irreversible, the movable shutter 5 is partially destroyed by the action of the magnetic field.
- the inhomogeneous magnetic field is generated here by a suitable magnet, for example an electromagnet 26. Therefore, if the state change is to be brought about, the reaction vessel 1 does not have to be moved, but rather the electromagnet 26 can simply be switched on.
- Reaction vessel 1 shown is a 0.2 ml standard PCR tube (Molecular Bioproducts, Inc., San Diego, Calif., USA) and corresponds to that described in US Pat FIG. 3 shown embodiment.
- the lid 3 of the reaction vessel 1 continues on the inside in a cylindrical projection 25, which is introduced when closing the lid 3 in the opening 2 of the reaction vessel 1 and contributes to a good sealing effect of the lid 3.
- a tube 4 is pressed, which has a length of 8 mm and is made of silicone.
- the inner diameter of the tube 4 is 2.45 mm, the outer diameter is 5 mm (quality SL601 and hardness 60 ° Shore, J. Lindemann GmbH, Helmstedt).
- the pressed-in the lid 4 end of the tube 4 is sealed by the lid 3.
- the still open end of the tube 4 is closed by the movable shutter 5 in the form of a Teflon coated ball 14 made of magnetic stainless steel (Nirosta material 1.4034) with a diameter of 2.54 mm.
- the movable shutter 5 can be removed again in an inhomogeneous magnetic field, which preferably has a gradient of at least 0.05 T / mm, particularly preferably> 0.1 T / mm.
- FIG. 14 An arrangement is shown with which such an inhomogeneous magnetic field can be generated.
- the ball can preferably be brought into a region of the reaction vessel through a small permanent magnet 9 (eg 5 mm diameter and 10 mm length), where it does not impair further investigation.
- a small permanent magnet 9 of NdFeB is shown, which is guided directly to the wall of the reaction vessel 1, so that the ball 14 approaches the permanent magnet 9.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
Die Erfindung betrifft ein Reaktionsgefäß mit einer ersten und einer zweiten Unterteilung, wobei in einem ersten Zustand des Reaktionsgefäßes die erste von der zweiten Unterteilung getrennt ist und in einem zweiten Zustand die erste mit der zweiten Unterteilung verbunden ist. Ferner ist die Verwendung des Reaktionsgefäßes in einem Verfahren zum Nachweis von Proteinen und/oder Nukleinsäuren und in einem Verfahren zur Vervielfältigung von Nukleinsäuren Teil der Erfindung. Die Erfindung umfasst ferner ein Verfahren zur Überführung des einen in den anderen der beiden Zustände des Reaktionsgefäßes.The invention relates to a reaction vessel having a first and a second subdivision, wherein in a first state of the reaction vessel, the first is separated from the second subdivision and in a second state, the first is connected to the second subdivision. Furthermore, the use of the reaction vessel in a method for the detection of proteins and / or nucleic acids and in a method for the amplification of nucleic acids is part of the invention. The invention further includes a method for converting the one into the other of the two states of the reaction vessel.
Die Patentanmeldung
Die Patentanmeldung
Das
In der Patentschrift
Die Patentanmeldung
Aus der internationalen Offenlegungsschrift
Die Patentschrift
Die Patentanmeldung
Ähnliche Vorrichtungen sind aus der japanischen Offenlegungsschrift
Der Erfindung liegt die Aufgabe zugrunde, ein verbessertes Reaktionsgefäß und eine Verwendung für ein solches verbessertes Reaktionsgefäß bereitzustellen. Zudem besteht eine Aufgabe der Erfindung darin, ein Verfahren unter Einsatz des verbesserten Reaktionsgefäßes zur Verfügung zu stellen.The invention has for its object to provide an improved reaction vessel and a use for such an improved reaction vessel. In addition, an object of the invention is to provide a method using the improved reaction vessel available.
Die Aufgabe wird erfindungsgemäß durch ein Reaktionsgefäß mit den Merkmalen der Ansprüche 1, 2 und 3 gelöst. Die Aufgabe wird zudem durch eine Verwendung des Reaktionsgefäßes in einem Verfahren zur Aufreinigung, Modifizierung und/oder zum Nachweis von Proteinen und/oder Nukleinsäuren gelöst. Eine weitere Lösung der erfindungsgemäßen Aufgabe besteht in einem Verfahren zur Überführung des einen in den anderen der beiden Zustände des Reaktionsgefäßes.The object is achieved by a reaction vessel with the features of
Das Reaktionsgefäß ist ein Gefäß, das dazu geeignet ist, dass eine Reaktion in ihm ausgeführt werden kann. Vorzugsweise ist das Reaktionsgefäß verschließbar, z. B. mit einem Deckel. Das bevorzugt verschlossene Reaktionsgefäß ist gegenüber seiner Umgebung flüssigkeitsdicht, besonders vorzugsweise ist es gasdicht.The reaction vessel is a vessel capable of reacting in it. Preferably, the reaction vessel is closable, z. B. with a lid. The preferred closed reaction vessel is liquid-tight with respect to its environment, particularly preferably it is gas-tight.
Das Reaktionsgefäß kann prinzipiell jede beliebige Form und Größe annehmen. Bevorzugt ist das Reaktionsgefäß im Wesentlichen zylindrisch, kreiszylindrisch oder konisch, besonders vorzugsweise weist das Reaktionsgefäß sowohl einen im Wesentlichen kreiszylindrischen Anteil als auch einen im Wesentlichen konischen Anteil auf. In einer anderen Ausführungsform hat das Reaktionsgefäß z.T. plane Wände, ins besondere z.B. eine rechteckige Grundfläche, wie bei handelsüblichen Küvetten. Vorzugsweise beträgt der Rauminhalt des Reaktionsgefäßes weniger als 500 ml, besonders vorzugsweise weniger als 100 ml, besonders vorzugsweise als 50 ml, besonders vorzugsweise weniger als 25 ml, besonders vorzugsweise weniger als 10 ml, besonders vorzugsweise weniger als 5 ml, besonders vorzugsweise weniger als 2,5 ml, besonders vorzugsweise kleiner als 1 ml, besonders vorzugsweise kleiner als 0,5 ml und ganz besonders vorzugsweise kleiner 0,25 ml. Zugleich ist der bevorzugte Rauminhalt des Reaktionsgefäßes größer als 0,05 ml, besonders bevorzugt ist er größer als 0,1 ml. Vorzugsweise ist das Reaktionsgefäß zumindest teilweise, besonders vorzugsweise in seiner Gesamtheit aus einem oder mehreren inerten Materialien wie Glas oder Kunststoff, vorzugsweise Polyethylen, Polypropylen oder Silikon, gefertigt. Darüber hinaus ist das Reaktionsgefäß vorzugsweise transparent, damit eine optische Inspektion des Öffnungsmechanismus bzw. des Öffnungserfolges möglich ist.The reaction vessel can in principle assume any shape and size. Preferably, the reaction vessel is substantially cylindrical, circular cylindrical or conical, more preferably, the reaction vessel has both a substantially circular cylindrical portion and a substantially conical portion. In another embodiment, the reaction vessel z.T. plane walls, in particular e.g. a rectangular base, as in commercial cuvettes. Preferably, the volume of the reaction vessel is less than 500 ml, more preferably less than 100 ml, more preferably less than 50 ml, more preferably less than 25 ml, more preferably less than 10 ml, most preferably less than 5 ml, most preferably less than 2 , 5 ml, particularly preferably less than 1 ml, more preferably less than 0.5 ml and most preferably less than 0.25 ml. At the same time, the preferred volume of the reaction vessel is greater than 0.05 ml, more preferably greater than 0 , 1 ml. Preferably, the reaction vessel is at least partially, particularly preferably in its entirety made of one or more inert materials such as glass or plastic, preferably polyethylene, polypropylene or silicone. In addition, the reaction vessel is preferably transparent, so that an optical inspection of the opening mechanism or the opening success is possible.
Das Reaktionsgefäß weist mindestens zwei Unterteilungen auf. In dem ersten Zustand ist die erste von der zweiten Unterteilung getrennt, sodass keine Flüssigkeit - vorzugsweise auch keine Gase - von einer der beiden Unterteilungen in die andere fließen kann. In dem zweiten Zustand ist die erste mit der zweiten Unterteilung verbunden und Flüssigkeiten - vorzugsweise auch Gase - können von einer der beiden Unterteilungen in die andere fließen.The reaction vessel has at least two subdivisions. In the first state, the first is separated from the second subdivision, so that no liquid - preferably also no gases - can flow from one of the two subdivisions into the other. In the second state, the first is connected to the second subdivision and liquids - preferably also gases - can flow from one of the two subdivisions into the other.
Eine Unterteilung kann an die andere angrenzen, eine kann die andere umfassen. Die Unterteilungen können auch voneinander beabstandet sein, so dass sie nicht aneinander angrenzen. Das verschlossene Reaktionsgefäß umschließt die Unterteilungen. Vorzugsweise sind die Unterteilungen im Reaktionsgefäß befestigt. Es ist jedoch auch möglich, dass eine oder beide Unterteilungen sich lose im Reaktionsgefäß befinden.One subdivision may adjoin the other, one may comprise the other. The partitions may also be spaced apart so that they do not abut each other. The sealed reaction vessel encloses the subdivisions. Preferably, the subdivisions are fixed in the reaction vessel. However, it is also possible that one or both subdivisions are loosely in the reaction vessel.
Beim Übergang vom ersten in den zweiten Zustand werden die Unterteilungen verbunden. Dies kann reversibel geschehen, sodass das Reaktionsgefäß vom zweiten wieder in den ersten Zustand überführt werden kann. Es ist jedoch auch möglich, dass der zweite Zustand irreversibel hergestellt wird, z. B. wenn eine Wand, die die erste von der zweiten Unterteilung trennt, bei der Zustandsänderung zerstört wird.In the transition from the first to the second state, the subdivisions are connected. This can be done reversibly, so that the reaction vessel from the second can be converted back into the first state. However, it is also possible that the second state is irreversibly produced, for. When a wall separating the first from the second partition is destroyed in the state change.
Das Reaktionsgefäß kann vorzugsweise noch weitere Unterteilungen aufweisen. Die weiteren Unterteilungen können in dem ersten und/oder zweiten Zustand mit der ersten und/oder der zweiten Unterteilung und/oder untereinander verbunden oder voneinander getrennt sein. Zudem kann vorzugsweise das Reaktionsgefäß noch weitere Zustände annehmen. In den weiteren Zuständen können vorzugsweise ein oder mehrere der weiteren Unterteilungen mit einer oder mehreren der weiteren Unterteilungen und/oder mit der ersten und/oder der zweiten Unterteilung verbunden oder von diesen getrennt sein.The reaction vessel may preferably have further subdivisions. The further subdivisions may be in the first and / or second state with the first and / or the second subdivision and / or interconnected or separated from each other. In addition, the reaction vessel may preferably assume further states. In the further states, one or more of the further subdivisions may preferably be connected to or separate from one or more of the further subdivisions and / or with the first and / or the second subdivision.
In dem erfindungsgemäßen Verfahren kann der erste und/oder zweite Zustand durch das Einwirken eines magnetischen Feldes in den anderen der beiden Zustände überführt werden. Durch Einwirken eines magnetischen Feldes kann die erste von der zweiten Unterteilung getrennt oder die erste mit der zweiten Unterteilung verbunden werden.In the method according to the invention, the first and / or second state can be converted into the other of the two states by the action of one magnetic field. By the action of a magnetic field, the first can be separated from the second subdivision or the first can be connected to the second subdivision.
Erfindungsgemäß kann das Reaktionsgefäß in einem Verfahren zur Aufreinigung, Modifizierung und/oder zum Nachweis von Proteinen und/oder Nukleinsäuren verwendet werden. Dabei kann eine erste Reaktion in der ersten Unterteilung ablaufen, die zunächst von der zweiten Unterteilung getrennt ist. Die erste Reaktion kann z. B. die Vermehrung von Nukleinsäuren mittels PCR oder die Aufreinigung von Nukleinsäuren oder Proteinen aus einer Probe sein. Durch Verbinden der beiden Unterteilungen können Reagenzien aus der zweiten Unterteilung zu der ersten Unterteilung hinzugefügt werden, sodass eine zweite Reaktion ablaufen kann. Die zweite Reaktion kann z. B. eine Nachweisreaktion auf eine spezifische Nukleinsäure oder ein spezifisches Protein sein. Auf diese Weise können Nukleinsäuren und Proteine besonders einfach und kontaminationsarm analysiert werden. Die Modifizierung kann beispielsweise in einer Verdauung einer Probe oder Markierung eines Proteins oder einer Nukleinsäure bestehen. Das Reaktionsgefäß kann erfindungsgemäß in einem Verfahren zur Vervielfältigung von Nukleinsäuren verwendet werden. Dabei sind vorzugsweise in der ersten Unterteilung lediglich ein Teil der für die Vervielfältigung benötigten Reagenzien vorhanden. Der übrige Teil ist vorzugsweise ausschließlich in der zweiten Unterteilung enthalten. Zur Ausführung der Vervielfältigung kann die erste Unterteilung mit der zweiten Unterteilung durch Einwirken eines magnetischen Feldes verbunden werden. Durch die getrennte Lagerung der beiden Teile ist eine höhere Haltbarkeit der Reagenzien erreichbar. Zudem können durch die gezielte An- oder Abwesenheit und die Konzentration der jeweiligen Reagenzien in jedem der beiden Teile die jeweils gewünschten Reaktionsbedingungen optimiert eingestellt werden.According to the invention, the reaction vessel can be used in a process for the purification, modification and / or detection of proteins and / or nucleic acids. In this case, a first reaction can take place in the first subdivision, which is initially separated from the second subdivision. The first reaction may, for. Example, the proliferation of nucleic acids by PCR or the purification of nucleic acids or proteins from a sample. By combining the two subdivisions, reagents from the second subdivision to the first subdivision so that a second reaction can proceed. The second reaction may, for. B. be a detection reaction to a specific nucleic acid or a specific protein. In this way, nucleic acids and proteins can be analyzed in a particularly simple and low-contamination manner. The modification may be, for example, in a digestion of a sample or label of a protein or a nucleic acid. The reaction vessel can be used according to the invention in a method for the amplification of nucleic acids. In this case, preferably only a part of the reagents required for the duplication are present in the first subdivision. The remaining part is preferably contained exclusively in the second subdivision. To perform the duplication, the first partition may be connected to the second partition by the action of a magnetic field. Due to the separate storage of the two parts a higher shelf life of the reagents can be achieved. In addition, the particular desired reaction conditions can be optimally adjusted by the targeted presence or absence and the concentration of the respective reagents in each of the two parts.
Die Erfindung enthält ferner ein Verfahren zur Zustandsänderung des Reaktionsgefäßes. In diesem Verfahren wird das Reaktionsgefäß einem magnetischen Feld ausgesetzt. Dabei ist vor dem Aussetzen entweder kein magnetisches Feld vorhanden oder ein zuvor vorhandenes magnetisches Feld wird beim Aussetzen in Ausrichtung und/oder Feldstärke verändert. Dies kann durch eine Bewegung des Reaktionsgefäßes relativ zu einem vorhandenen magnetischen Feld verwirklicht werden. Alternativ kann auch ein zeitlich nicht konstantes magnetisches Feld verwendet werden, z.B. kann zur Zustandsänderung ein magnetisches Feld mit einem Elektromagneten erzeugt oder verändert werden.The invention further includes a method for changing the state of the reaction vessel. In this process, the reaction vessel is exposed to a magnetic field. In this case, either no magnetic field is present before exposure or a previously existing magnetic field is changed when exposed to alignment and / or field strength. This can be accomplished by moving the reaction vessel relative to an existing magnetic field. Alternatively, a non-constant magnetic field may be used, e.g. For changing the state, a magnetic field can be generated or changed by means of an electromagnet.
Durch die Erfindung ist es vorteilhafterweise möglich, zwei unterschiedliche Flüssigkeiten in zwei Unterteilungen eines Reaktionsgefäßes aufzubewahren, ohne dass sich die Flüssigkeiten vermischen. Zu gegebener Zeit kann dann, vorzugsweise ohne dass das Gefäß geöffnet werden muss, durch Einwirken eines magnetischen Feldes, eine Unterteilung mit der anderen verbunden werden, so dass sich die beiden Flüssigkeiten miteinander vermischen können. So sind zweistufige Nachweismethoden ohne ein zwischenzeitliches Öffnen des Reaktionsgefäßes möglich, wodurch auf Arbeitsschritte eingespart und Kontaminationen vermieden werden können. Der Verzicht auf die Öffnung des Reaktionsgefäßes zur Vermischung der Flüssigkeiten kann die Automatisierung von Reaktionen vereinfachen. Im Stand der Technik werden Wachse zum Trennen von zwei Flüssigkeiten in einem Reaktionsgefäß verwendet, die zum Vermischen der Flüssigkeiten erhitzt werden; sobald das Wachs erkaltet und somit erhärtet ist dann keine Vermischung mehr möglich bzw. stark erschwert. Erfindungsgemäß kann ein inertes Material zur Trennung der Flüssigkeiten in den Unterteilungen verwendet werden, zur Vermischung der Flüssigkeiten ist kein Erhitzen nötig, wodurch eine Beeinträchtigung von Reaktionen vermieden werden kann. Zudem ermöglicht die Erfindung die Herstellung eines konstruktiv einfachen, günstig zu fertigenden Reaktionsgefäßes.By means of the invention, it is advantageously possible to store two different liquids in two subdivisions of a reaction vessel without the liquids mixing. In due course, then, preferably without the vessel having to be opened, by the action of a magnetic field, one partition can be connected to the other so that the two liquids can mix with one another. Thus, two-step detection methods without an interim opening of the reaction vessel are possible, which saves on work steps and contamination can be avoided. The abandonment of the opening of the reaction vessel for mixing the liquids can be the automation of Simplify reactions. In the prior art, waxes are used to separate two liquids in a reaction vessel, which are heated to mix the liquids; as soon as the wax cools down and hardens, mixing is no longer possible or greatly impeded. According to the invention, an inert material can be used to separate the liquids in the subdivisions, no mixing is necessary for mixing the liquids, whereby an impairment of reactions can be avoided. In addition, the invention makes it possible to produce a structurally simple, inexpensive reaction vessel.
Vorzugsweise sind im ersten Zustand die erste und zweite Unterteilung durch einen beweglichen Verschluss voneinander getrennt. Durch eine geeignete Bewegung des beweglichen Verschlusses kann die erste mit der zweiten Unterteilung verbunden werden. Der bewegliche Verschluss kann prinzipiell eine beliebige Form annehmen, vorzugsweise ist der bewegliche Verschluss jedoch sphärisch oder zylindrisch geformt. Durch die Wahl eines geeigneten beweglichen Verschlusses kann die Verbindung der ersten mit der zweiten Unterteilung reversibel gestaltet werden. So kann bei geeigneter Ausführung auch erreicht werden, dass durch eine Bewegung des beweglichen Verschlusses eine Trennung von zunächst verbundenen Unterteilungen herbeigeführt werden kann. Bewegung des beweglichen Verschlusses, die zur Zustandsänderung des Reaktionsgefäßes führt, kann in einer translatorischen oder rotatorischen Bewegung oder aus einer Kombination der beiden Bewegungsarten bestehen. Der bewegliche Verschluss kann aus einem einzelnen oder mehreren Elementen bestehen. In einer Ausführungsform ist der bewegliche Verschluss eine Kugel, die die erste Unterteilung gegenüber der zweiten Unterteilung abschließt. Ein einteiliger beweglicher Verschluss kann vorteilhafterweise besonders einfach produziert werden.Preferably, in the first state, the first and second subdivisions are separated by a movable shutter. By suitable movement of the movable shutter, the first can be connected to the second partition. The movable closure may in principle take any shape, but preferably the movable closure is spherical or cylindrical in shape. By choosing a suitable movable closure, the connection of the first with the second subdivision can be made reversible. Thus, with a suitable design can also be achieved that a separation of initially associated subdivisions can be brought about by a movement of the movable closure. Movement of the movable shutter, which leads to the change of state of the reaction vessel, may consist of a translatory or rotational movement or a combination of the two types of movement. The movable shutter may consist of a single or multiple elements. In one embodiment, the movable shutter is a ball that closes off the first partition from the second partition. A one-piece movable closure can advantageously be produced particularly easily.
In einer weiteren Ausführungsform besteht der bewegliche Verschluss aus einer Klappe und einem Riegel. Die Klappe kann die beiden Unterteilungen gegeneinander abgrenzen, der Riegel kann die Klappe arretieren. Um die Zustandsänderung herbeizuführen muss in dieser Ausführungsform zunächst der Riegel bewegt werden, so dass die Klappe freigegeben wird. Erst dann kann die Klappe bewegt werden. Mit einem solchen zweiteiligen beweglichen Verschluss kann verhindert werden, dass die Zustandsänderung z.B. während einer optionalen Zentrifugation oder durch Stöße unbeabsichtigt erzeugt wird, z.B. durch einen ungepolsterten Fall auf den Boden. Eine solche unbeabsichtigte, vorzeitige Zustandsänderung kann vorzugsweise durch den erfindungsgemäßen Riegel auch in solchen Fällen verhindert werden, in denen sich in der verschlossenen Unterteilung ein hoher Überdruck aufbaut (sei es durch eine hohe Temperatur, durch die Verwendung sehr flüchtiger Chemikalien, durch reaktionsbedingte Gasbildung oder durch geringen Außendruck z.B. während des Transports).In a further embodiment, the movable closure consists of a flap and a latch. The flap can separate the two divisions against each other, the latch can lock the flap. In order to bring about the state change, in this embodiment, first of all, the latch must be moved, so that the flap is released. Only then can the flap be moved. With such a Two-piece movable closure can be prevented that the change of state, for example, during an optional centrifugation or shocks generated accidentally, for example, by an un-padded case on the ground. Such an unintentional, premature change of state can preferably be prevented by the bar according to the invention also in cases in which a high pressure builds up in the closed subdivision (be it by a high temperature, by the use of very volatile chemicals, by reaction-induced gas formation or by low external pressure eg during transport).
In einer bevorzugten Ausführungsform enthält der bewegliche Verschluss eine magnetische Komponente. Besonders vorzugsweise besteht der bewegliche Verschluss aus der magnetischen Komponente. Die bevorzugte magnetische Komponente ist zylindrisch oder sphärisch. Durch das Einwirken des magnetischen Feldes kann die magnetische Komponente des beweglichen Verschlusses bewegt werden. In einem inhomogenen magnetischen Feld kann sich die magnetische Komponente von einem Ort der schwächeren zu einem Ort der stärkeren magnetischen Feldstärke bewegen. Alternativ oder zusätzlich kann sich die magnetische Komponente des beweglichen Verschlusses durch Einwirken des magnetischen Feldes entlang der Feldlinie des magnetischen Feldes ausrichten. Dies kann sowohl in einem inhomogenen als auch in einem homogenen magnetischen Feld geschehen. Durch die magnetische Komponente des beweglichen Verschlusses kann die Zustandsänderung durch Einwirken des magnetischen Feldes technisch besonders einfach erreicht werden. Der bewegliche Verschluss kann z.B. aus einer magnetischen Komponente (z.B. aus einem ferromagnetischen Material) bestehen, die zur besseren Dichtwirkung und/oder zum Korrosionsschutz mit einem Kunststoffüberzug versehen ist. Alternativ kann der bewegliche Verschluss z.B. auch aus einem nichtmagnetischen Körper bestehen, der mit einer ferromagnetischen Schicht versehen ist, die die magnetische Komponente darstellt.In a preferred embodiment, the movable shutter includes a magnetic component. Particularly preferably, the movable closure consists of the magnetic component. The preferred magnetic component is cylindrical or spherical. By the action of the magnetic field, the magnetic component of the movable shutter can be moved. In an inhomogeneous magnetic field, the magnetic component may move from a weaker location to a stronger magnetic field location. Alternatively or additionally, the magnetic component of the movable shutter can be aligned by the action of the magnetic field along the field line of the magnetic field. This can be done both in an inhomogeneous as well as in a homogeneous magnetic field. Due to the magnetic component of the movable shutter, the state change can be achieved particularly easily by the action of the magnetic field. The movable shutter may e.g. consist of a magnetic component (e.g., a ferromagnetic material) provided with a plastic coating for better sealing and / or corrosion protection. Alternatively, the movable shutter may be e.g. Also, consist of a non-magnetic body, which is provided with a ferromagnetic layer, which represents the magnetic component.
Der bewegliche Verschluss mit der magnetischen Komponente kann in eine Trennwand zwischen der ersten und zweiten Unterteilung eingebracht sein. Durch Einwirken des magnetischen Feldes kann sich die magnetische Komponente so bewegen und/oder sich so ausrichten, dass eine Verbindung zwischen erster und zweiter Unterteilung entsteht. Vorzugsweise bestehen Teile oder die gesamte Trennwand zwischen erster und zweiter Unterteilung aus der magnetischen Komponente. Auf diese Weise kann das Reaktionsgefäß mit besonders geringem Materialaufwand hergestellt werden.The movable shutter with the magnetic component may be incorporated in a partition wall between the first and second divisions. By acting on the magnetic field, the magnetic component can move and / or align so as to form a connection between the first and second subdivisions. Preferably, parts or the entire partition wall between the first and second Subdivision of the magnetic component. In this way, the reaction vessel can be produced with a particularly low cost of materials.
Die magnetische Komponente kann ein ferromagnetisches Material enthalten oder aus einem ferromagnetischen Material bestehen. Vorzugsweise enthält die magnetische Komponente Eisen. Durch die Verwendung von Eisen in der magnetischen Komponente können die Herstellungskosten des Reaktionsgefäßes besonders niedrig gehalten werden.The magnetic component may include a ferromagnetic material or be made of a ferromagnetic material. Preferably, the magnetic component contains iron. By using iron in the magnetic component, the manufacturing cost of the reaction vessel can be kept particularly low.
In einer Ausführungsform ist die magnetische Komponente zumindest teilweise permanentmagnetisch. Vorzugsweise besteht die magnetische Komponente vollständig aus einem Permanentmagneten. Durch die Verwendung eines Permanentmagneten ist es möglich, dass ein einwirkendes magnetisches Feld eine größere Kraft auf die magnetische Komponente ausüben kann, als wenn die magnetische Komponente lediglich aus einem ferromagnetischen, nicht magnetisierten Material besteht. Zudem kann durch einen Permanentmagneten erreicht werden, dass sich die magnetische Komponente in einem magnetischen Feld ausrichtet. Zum Beispiel kann sich die magnetische Komponente in einem magnetischen Feld entlang der Feldlinien ausrichten. Bei geeigneter Ausgestaltung entsteht durch die Ausrichtung entlang der Feldlinien eine Drehbewegung der magnetischen Komponente, durch die die erste und die zweite Unterteilung miteinander verbunden oder voneinander getrennt werden können.In one embodiment, the magnetic component is at least partially permanent magnetic. Preferably, the magnetic component consists entirely of a permanent magnet. By using a permanent magnet, it is possible for an applied magnetic field to exert a greater force on the magnetic component than if the magnetic component consists solely of a ferromagnetic, non-magnetized material. In addition, can be achieved by a permanent magnet, that aligns the magnetic component in a magnetic field. For example, the magnetic component may align in a magnetic field along the field lines. In a suitable embodiment, the alignment along the field lines produces a rotational movement of the magnetic component, by means of which the first and the second subdivision can be connected to one another or separated from one another.
Erfindungsgemäß ist es bevorzugt, dass der bewegliche Verschluss im ersten oder zweiten Zustand kraftschlüssig mit dem Reaktionsgefäß verbunden ist. Dabei liegt vorzugsweise eine Fläche des beweglichen Verschlusses an einer Fläche des Reaktionsgefäßes so an, dass eine Kraft entlang der Flächennormale eine ausreichende Reibung zwischen den Flächen erzeugt, um den beweglichen Verschluss in dem Reaktionsgefäß festzuhalten. Vorzugsweise kann ein Einwirken des magnetischen Feldes eine ausreichende Kraft auf die magnetische Komponente des beweglichen Verschlusses ausüben, um die Reibung der beiden genannten Flächen aufeinander zu überwinden, sodass sich der bewegliche Verschluss im Reaktionsgefäß bewegt und die erste und zweite Unterteilung miteinander verbunden oder voneinander getrennt werden können. Prinzipiell können der bewegliche Verschluss und die angrenzenden Teile des Reaktionsgefäßes eine beliebige Form annehmen. In einer Ausführungsform besteht der bewegliche Verschluss aus einem magnetischen rotationssymmetrischen Körper, vorzugsweise aus einem permanentmagnetischen Kreiszylinder oder einer permanentmagnetischen Kugel. Der Durchmesser des Körpers beträgt dabei vorzugsweise zwischen 0,1 und 10 mm, besonders vorzugsweise zwischen 1 und 6 mm und ganz besonders vorzugsweise zwischen 2 und 4 mm. Der Körper kann in einen flexiblen Schlauch eingebracht werden, der an einem Ende verschlossen ist. In einer Ausführungsform ist der Schlauch aus Silikon gefertigt. Bei der Ausführung des Körpers als Kugel oder Kreiszylinder beträgt das Verhältnis des Durchmessers des Körpers zu dem Innendurchmesser des Schlauches vorzugsweise 1:1, besonders vorzugsweise 1,01:1, besonders vorzugsweise 1,03:1, besonders vorzugsweise 1,05:1, besonders vorzugsweise 1,1:1, besonders vorzugsweise 1,2:1, besonders vorzugsweise 1,3:1, und ganz besonders vorzugsweise 1,5:1. Vorzugsweise wird der Schlauch beim Einbringen des Körpers so gedehnt, dass eine dichte Verbindung zwischen Schlauch und Körper entsteht. Der Schlauch hat dabei vorzugsweise eine Wandstärke zwischen 0,1 und 5 mm, besonders vorzugsweise zwischen 0,5 und 3 mm und ganz besonders vorzugsweise zwischen 0,7 und 1,5 mm. In dem verschlossenen Endes des Schlauches befindet sich die erste Unterteilung, in der sich eine Flüssigkeit befinden kann. Durch das Einwirken eines magnetischen Feldes kann die Haftreibung, die den Körper in dem Schlauch hält, überwunden werden, sodass der Körper aus dem Schlauch entfernt wird und die erste Unterteilung mit einer den Schlauch umfassenden zweiten Unterteilung verbunden wird. Durch eine kraftschlüssige Verbindung des beweglichen Verschlusses mit dem Reaktionsgefäß kann die Erfindung konstruktiv besonders einfach ausgeführt werden.According to the invention it is preferred that the movable closure in the first or second state is non-positively connected to the reaction vessel. Preferably, an area of the movable shutter on a surface of the reaction vessel is such that a force along the surface normal generates sufficient friction between the surfaces to hold the movable shutter in the reaction vessel. Preferably, exposure to the magnetic field may exert sufficient force on the magnetic component of the movable shutter to overcome the friction of the two surfaces, such that the movable shutter moves in the reaction vessel and the first and second subdivisions are connected or disconnected can. In principle, the movable closure and the adjacent parts of the reaction vessel can take any shape. In one embodiment, the movable closure of a magnetic rotationally symmetrical body, preferably of a permanent magnetic circular cylinder or a permanent magnetic ball. The diameter of the body is preferably between 0.1 and 10 mm, more preferably between 1 and 6 mm and most preferably between 2 and 4 mm. The body can be inserted into a flexible tube which is closed at one end. In one embodiment, the tube is made of silicone. In the embodiment of the body as a sphere or circular cylinder, the ratio of the diameter of the body to the inner diameter of the tube is preferably 1: 1, particularly preferably 1.01: 1, particularly preferably 1.03: 1, particularly preferably 1.05: 1, more preferably 1.1: 1, more preferably 1.2: 1, most preferably 1.3: 1, and most preferably 1.5: 1. Preferably, the hose is stretched during insertion of the body so that a tight connection between the hose and the body is formed. The tube preferably has a wall thickness between 0.1 and 5 mm, more preferably between 0.5 and 3 mm, and most preferably between 0.7 and 1.5 mm. In the closed end of the tube is the first subdivision, in which a liquid can be located. By the action of a magnetic field, the stiction holding the body in the tube can be overcome so that the body is removed from the tube and the first partition is connected to a second partition comprising the tube. By a non-positive connection of the movable closure with the reaction vessel, the invention can be structurally particularly simple.
In einer bevorzugten Ausführungsform der Erfindung ist der bewegliche Verschluss im ersten oder zweiten Zustand formschlüssig mit dem Reaktionsgefäß verbunden. Vorzugsweise ist ein erster Anteil des beweglichen Verschlusses so angeordnet, dass er einem zweiten Anteil des beweglichen Verschlusses den Weg versperrt, der zur Verbindung der ersten Unterteilung mit der zweiten Unterteilung zurückgelegt werden muss. Vorzugsweise ist der erste Anteil ein Riegel, der den zweiten Anteil so versperrt, dass keine Verbindung zwischen der ersten und der zweiten Unterteilung entstehen kann. Besonders bevorzugt ist der zweite Anteil ein schwenkbarer Deckel und der erste Anteil ein Riegel, der die magnetische Komponente enthält. Durch Einwirken des magnetischen Feldes kann so der Riegel verschoben werden, wodurch der Deckel nicht mehr versperrt wird und geöffnet werden kann. Die Öffnung des Deckels kann durch ein geeignetes elastisches Element, z.B. eine Stahlfeder, oder bei einem entsprechenden Material des Deckels durch Vorspannung des Deckels erleichtert werden. Die formschlüssige Verbindung des beweglichen Verschlusses ermöglicht eine besonders sichere Trennung der ersten von der zweiten Unterteilung im ersten Zustand. Ein versehentliches Lösen des beweglichen Verschlusses, z.B. bei einem optionalen Zentrifugationsschritt, kann so vermieden werden.In a preferred embodiment of the invention, the movable closure in the first or second state is positively connected to the reaction vessel. Preferably, a first portion of the movable shutter is arranged to obstruct a second portion of the movable shutter the path that must be traveled to connect the first subdivision with the second subdivision. Preferably, the first portion is a bar which blocks the second portion so that no connection between the first and the second partition can occur. Particularly preferably, the second portion is a hinged lid and the first portion is a bar containing the magnetic component. By acting on the magnetic field so the bolt can be moved, so that the lid is no longer locked and can be opened. The opening of the lid can by a suitable elastic element, such as a steel spring, or be relieved at a corresponding material of the lid by biasing the lid. The positive connection of the movable closure allows a particularly secure separation of the first from the second subdivision in the first state. An accidental release of the movable closure, for example in an optional centrifugation step, can be avoided.
In einer bevorzugten Ausführungsform der Erfindung ist der bewegliche Verschluss im ersten oder zweiten Zustand stoffschlüssig mit dem Reaktionsgefäß verbunden. Dabei ist der bewegliche Verschluss irreversibel mit dem übrigen Reaktionsgefäß zusammengefügt. Eine Zustandsänderung, in der die erste und zweite Unterteilung miteinander verbunden werden, kann nur durch Zerstörung von Teilen des beweglichen Verschlusses erreicht werden. In dieser Ausführung kann der bewegliche Verschluss z. B. in einer Folie bestehen, mit der die erste Unterteilung verschlossen ist. Eine in der Folie vorhandene magnetische Komponente kann durch Einwirken des magnetischen Feldes eine Kraft auf die Folie übertragen, die ausreicht, um die Folie zu zerstören und dadurch die erste mit der zweiten Unterteilung zu verbinden. Vorzugsweise kann der bewegliche Verschluss - bis auf die magnetische Komponente - einteilig mit dem übrigen Reaktionsgefäß gefertigt werden, wobei vorzugsweise Sollbruchstellen an dem beweglichen Verschluss angebracht sind, sodass durch Einwirken des magnetischen Feldes die magnetische Komponente auf den beweglichen Verschluss eine Kraft ausübt, die zur Zerstörung der Sollbruchstellen und damit zur Zustandsänderung des Reaktionsgefäßes führt.In a preferred embodiment of the invention, the movable closure in the first or second state is firmly bonded to the reaction vessel. In this case, the movable closure is irreversibly joined to the rest of the reaction vessel. A state change in which the first and second subdivisions are connected together can only be achieved by destroying parts of the movable shutter. In this embodiment, the movable closure z. B. consist of a film with which the first subdivision is closed. A magnetic component present in the film can, by the action of the magnetic field, transmit a force to the film sufficient to destroy the film and thereby connect the first to the second subdivision. Preferably, the movable closure - can be made in one piece with the rest of the reaction vessel except for the magnetic component, preferably predetermined breaking points are attached to the movable closure, so that by the action of the magnetic field, the magnetic component on the movable closure exerts a force to destroy the predetermined breaking points and thus the state change of the reaction vessel leads.
In einer bevorzugten Ausführungsform wird zumindest eine der Unterteilungen zumindest teilweise von einem Deckel des Reaktionsgefäßes begrenzt. Bei geöffnetem Deckel kann so Reaktionsflüssigkeit in zumindest eine der Unterteilungen eingefüllt werden. Nach Schließen des Deckels ist sind vorzugsweise beide Unterteilungen zu allen Seiten hin verschlossen. In dieser Ausführungsform ist das Einfüllen von Flüssigkeiten besonders einfach.In a preferred embodiment, at least one of the subdivisions is at least partially bounded by a cover of the reaction vessel. With the lid open, the reaction liquid can be filled into at least one of the subdivisions. After closing the lid both subdivisions are preferably closed on all sides. In this embodiment, the filling of liquids is particularly simple.
In einer bevorzugten Ausführungsform der Erfindung ist das Reaktionsgefäß mit zumindest einer der beiden Unterteilungen einstückig gefertigt. Vorzugsweise sind beide Unterteilungen mit dem Reaktionsgefäß einstückig gefertigt. Auch der bewegliche Verschluss kann vorzugsweise mit dem übrigen Reaktionsgefäß einstückig hergestellt sein. In dieser Fertigungsart kann das Reaktionsgefäß besonders günstig produziert werden.In a preferred embodiment of the invention, the reaction vessel is manufactured in one piece with at least one of the two subdivisions. Preferably, both subdivisions are made in one piece with the reaction vessel. Also the moving one Closure may preferably be made in one piece with the rest of the reaction vessel. In this type of production, the reaction vessel can be produced particularly cheap.
In einer bevorzugten Ausführungsform befindet sich zumindest eine der beiden Unterteilungen in einem im Reaktionsgefäß beweglichen, hohlen Körper. Vorzugsweise ist dieser hohle Körper dabei nicht im Reaktionsgefäß befestigt, sondern befindet sich lose im Reaktionsgefäß. Eine solche Unterteilung kann z. B. als Schlauch ausgeführt werden, der auf einer Seite durch den beweglichen Verschluss und auf der anderen Seite durch eine feste Wand verschlossen ist und nicht am übrigen Reaktionsgefäß befestigt ist. So ist es möglich, dass eine bereits verschlossene Unterteilung in das Reaktionsgefäß eingeführt wird, was bei der Verwendung von besonders volatilen Reagenzien von Vorteil sein kann.In a preferred embodiment, at least one of the two subdivisions is located in a hollow body which is movable in the reaction vessel. Preferably, this hollow body is not attached in the reaction vessel, but is located loosely in the reaction vessel. Such a subdivision may, for. B. are designed as a hose which is closed on one side by the movable closure and on the other side by a solid wall and is not attached to the rest of the reaction vessel. Thus, it is possible that an already closed subdivision is introduced into the reaction vessel, which may be advantageous when using particularly volatile reagents.
In dem erfindungsgemäßen Verfahren zur Zustandsänderung des Reaktionsgefäßes wird vorzugsweise ein inhomogenes magnetisches Feld zur Zustandsänderung verwendet. Die magnetische Komponente kann sich dabei in dem inhomogenen magnetischen Feld von Zonen mit niedriger magnetischer Feldstärke zu höherer magnetischer Feldstärke bewegen, wobei vorzugsweise die erste Unterteilung mit der zweiten Unterteilung verbunden wird oder diese beiden voneinander getrennt werden. Zudem kann bei einer permanentmagnetischen magnetischen Komponente sich diese im inhomogenen Magnetfeld in Richtung der Feldlinien ausrichten und so die Zustandsänderung erreicht werden. Alternativ wird die Zustandsänderung des Reaktionsgefäßes durch ein homogenes Magnetfeld herbeigeführt. Dabei ist vorzugsweise die magnetische Komponente permanentmagnetisch, so dass sie sich in dem magnetischen Feld dreht und entlang der Feldlinien ausrichtet. Die magnetische Komponente kann so angebracht werden, dass durch ihre Drehung der bewegliche Verschluss geöffnet wird. Z.B. kann der bewegliche Verschluss aus einer magnetisierten Kugel bestehen, die eine Bohrung aufweist. Die Kugel kann drehbar in einem Lager befestigt sein, so dass in einer Ausrichtung der Kugel die Bohrung durch das Lager dichtend verschlossen ist. Durch das Einwirken eines geeigneten homogenen magnetischen Feldes kann sich die Ausrichtung der permanentmagnetischen Kugel ändern, so dass die Bohrung nicht mehr verdeckt ist und die erste mit der zweiten Unterteilung über die Bohrung der Kugel verbunden wird.In the method according to the invention for changing the state of the reaction vessel, preference is given to using an inhomogeneous magnetic field for the change of state. The magnetic component may thereby move in the inhomogeneous magnetic field of zones of low magnetic field strength to higher magnetic field strength, wherein preferably the first subdivision is connected to the second subdivision or these two are separated from each other. In addition, in the case of a permanent magnetic magnetic component, these can be aligned in the direction of the field lines in the inhomogeneous magnetic field and the state change can thus be achieved. Alternatively, the state change of the reaction vessel is brought about by a homogeneous magnetic field. In this case, the magnetic component is preferably permanently magnetic, so that it rotates in the magnetic field and aligns along the field lines. The magnetic component can be mounted so that the movable shutter is opened by its rotation. For example, the movable shutter may consist of a magnetized ball having a bore. The ball may be rotatably mounted in a bearing, so that in an orientation of the ball, the bore is sealed by the bearing. By the action of a suitable homogeneous magnetic field, the orientation of the permanent magnetic sphere may change so that the bore is no longer obscured and the first is connected to the second subdivision via the bore of the sphere.
Vorteilhafterweise kann die Verbindung der Unterteilungen auf diese Weise reversibel gestaltet werden.Advantageously, the connection of the subdivisions can be made reversible in this way.
Vorzugsweise geschieht die Zustandsänderung des Reaktionsgefäßes unter Verwendung eines Permanentmagneten oder eines Elektromagneten. Durch die Verwendung eines Permanentmagneten kann die Zustandsänderung des Reaktionsgefäßes ohne ein elektrisches Gerät herbeigeführt werden. Dies ist insbesondere bei einer mobilen Verwendung der Reaktionsgefäße von Vorteil. Ein Elektromagnet kann das zur Zustandsänderung nötige magnetische Feld bei relativ geringer Baugröße erzeugen. Da durch den Elektromagneten das magnetische Feld nach Belieben an- und ausgeschaltet werden kann, ist keine Relativbewegung des Reaktionsgefäßes zum Elektromagneten nötig, um die Zustandsänderung auszulösen. Das Reaktionsgefäß kann vielmehr fest an einem Ort platziert werden und das magnetische Feld zu geeigneter Zeit angeschaltet werden.Preferably, the state change of the reaction vessel is done using a permanent magnet or an electromagnet. By using a permanent magnet, the state change of the reaction vessel can be brought about without an electric device. This is particularly advantageous for mobile use of the reaction vessels. An electromagnet can generate the magnetic field necessary for the change of state in a relatively small size. Since the magnetic field can be switched on and off as desired by the electromagnet, no relative movement of the reaction vessel to the electromagnet is necessary to trigger the change of state. Rather, the reaction vessel can be placed firmly in place and the magnetic field switched on at the appropriate time.
In einer bevorzugten Ausführung geschieht die Zustandsänderung des Reaktionsgefäßes unter Verwendung zweier Permanent- und/oder Elektromagnete, die derart angeordnet sind, dass zwischen den Permanent- und/oder Elektromagneten ein Spalt entsteht. Die Erfinder haben durch Simulationen herausgefunden, dass durch eine derartige Anordnung von Magneten ein inhomogenes Feld erzeugt werden kann, dessen räumliche Änderung der magnetischen Flussdichte ausreichend ist, um die für die Zustandsänderung des Reaktionsgefäßes nötige Kraft auf die magnetische Komponente bereitzustellen. Dabei ist der Betrag des Gradienten des Magnetfeldes vorzugsweise zwischen 0,001 und 10 T/mm, besonders vorzugsweise zwischen 0,01 und 1 T/mm und ganz besonders vorzugsweise zwischen 0,03 und 0,3 T/mm.In a preferred embodiment, the state change of the reaction vessel using two permanent and / or electromagnets, which are arranged such that between the permanent and / or electromagnets, a gap is formed. The inventors have found through simulations that such an array of magnets can create an inhomogeneous field whose spatial variation in magnetic flux density is sufficient to provide the necessary force for the change in state of the reaction vessel to the magnetic component. In this case, the magnitude of the magnetic field gradient is preferably between 0.001 and 10 T / mm, more preferably between 0.01 and 1 T / mm and most preferably between 0.03 and 0.3 T / mm.
Sobald die Zustandsänderung eine Verbindung der Unterteilungen erlaubt, können sich vorzugsweise die in den Unterteilungen vorgelegten Substanzen vermischen bzw. miteinander reagieren. Hierbei geraten die Substanzen vorzugsweise nach dem Herstellen der Verbindung aufgrund der Schwerkraft in Kontakt. Je nach Beschaffenheit der Substanzen (insbesondere in Hinblick auf Oberflächenspannung und Viskosität), sowie je nach Beschaffenheit (u.a. Form, Größe, Oberfläche) der zuvor verschlossenen Unterteilung kann es erforderlich sein, das Reaktionsgefäß zu schütteln, zu vibrieren oder in geeigneter Form einer Zentrifugation auszusetzen, damit die Flüssigkeit herausfließt.As soon as the state change permits a combination of the subdivisions, the substances presented in the subdivisions may preferably mix or react with one another. In this case, the substances preferably come into contact after the connection has been established by gravity. Depending on the nature of the substances (in particular with regard to surface tension and viscosity), and depending on the nature (including shape, size, surface) of the previously closed subdivision, it may be necessary to shake the reaction vessel, vibrate or suspend in a suitable form of centrifugation so that the liquid flows out.
Bei wässrigen Lösungen ist hierfür häufig eine Zentrifugalbeschleunigung von vorzugsweise weniger als 10.000 g (g = 9,81 m/s2), besonders vorzugsweise weniger als 1000 g, besonders vorzugsweise von weniger als 100 g und ganz besonders weniger als 10 g erforderlich. Es kann ausreichen, das Reaktionsgefäß in die Hand zu nehmen und eine geeignete, kreisförmige schnelle Bewegung der Hand auszuführen.For aqueous solutions, this often requires a centrifugal acceleration of preferably less than 10,000 g (g = 9.81 m / s 2 ), more preferably less than 1000 g, more preferably less than 100 g and most preferably less than 10 g. It may be sufficient to pick up the reaction vessel and make a suitable circular rapid movement of the hand.
Es zeigen:
-
ein Reaktionsgefäß mit einem kreiszylindrischen beweglichen Verschluss;Figur 1 -
einen Versuchsaufbau, in dem das Reaktionsgefäß durch ein magnetisches Feld bewegt wird;Figur 2 -
ein Reaktionsgefäß mit einem sphärischen beweglichen Verschluss und einer am Deckel befestigten Unterteilung;Figur 3 -
ein Reaktionsgefäß in einer weiteren Ausführungsform mit einem sphärischen beweglichen Verschluss und einer am Deckel befestigten Unterteilung;Figur 4 -
Figur 5 ein Reaktionsgefäß mit einem sphärischen beweglichen Verschluss und einer deckelnahen Unterteilung im Reaktionsgefäß; -
ein Reaktionsgefäß mit einer Trennwand, die vom Deckel aus nur zu einer Unterteilung direkten Zugang erlaubt;Figur 6 -
ein Reaktionsgefäß mit einer Trennwand entlang einer Symmetrieebene des Reaktionsgefäßes;Figur 7 -
Figur 8 das erfindungsgemäße Reaktionsgefäß mit einer im Reaktionsgefäß frei beweglichen Unterteilung mit Abstandshaltern; -
in einer schematischen Darstellung den auf Trägheit basierenden Öffnungsvorgang einer frei beweglichen Unterteilung;Figur 9 -
ein Reaktionsgefäß, in dem die Trennung der Unterteilungen durch einen sphärischen beweglichen Verschluss, der in einem konischen Reaktionsgefäß aufliegt, verwirklicht ist;Figur 10 -
ein Reaktionsgefäß, in dem die Unterteilungen durch eine Ring geschaffen sind, in der sich ein kreiszylindrischer beweglicher Verschluss befindet;Figur 11 -
Figur 12 das erfindungsgemäße Reaktionsgefäß, in dem eine Unterteilung am Deckel angebracht ist und der bewegliche Verschluss eine Folie enthält, die bei der Zustandsänderung durch eine sphärische magnetische Komponente zerstört wird; -
Figur 13 eine Fotografie des Reaktionsgefäßes in der in gezeigten Ausführungsform;Figur 3 -
eine Fotografie des Versuchsaufbaus ausFigur 14Figur 2
und -
eine Fotografie eines Reaktionsgefäßes mit seitlich durch einen Permanentmagneten gesichertem beweglichen Verschluss.Figur 15
-
FIG. 1 a reaction vessel with a circular cylindrical movable closure; -
FIG. 2 a test setup in which the reaction vessel is moved by a magnetic field; -
FIG. 3 a reaction vessel having a spherical movable shutter and a partition attached to the lid; -
FIG. 4 a reaction vessel in a further embodiment with a spherical movable closure and a partition attached to the lid; -
FIG. 5 a reaction vessel with a spherical movable closure and a lid-like subdivision in the reaction vessel; -
FIG. 6 a reaction vessel with a dividing wall which allows direct access from the lid to a subdivision only; -
FIG. 7 a reaction vessel having a partition wall along a plane of symmetry of the reaction vessel; -
FIG. 8 the reaction vessel according to the invention with a freely movable in the reaction vessel subdivision with spacers; -
FIG. 9 in a schematic representation of the inertia-based opening operation of a freely movable subdivision; -
FIG. 10 a reaction vessel in which the separation of the partitions is realized by a spherical movable shutter which rests in a conical reaction vessel; -
FIG. 11 a reaction vessel in which the partitions are provided by a ring in which a circular cylindrical movable shutter is located; -
FIG. 12 the reaction vessel according to the invention, in which a subdivision is attached to the lid and the movable shutter contains a film which is destroyed in the state change by a spherical magnetic component; -
FIG. 13 a photograph of the reaction vessel in theFIG. 3 embodiment shown; -
FIG. 14 a photograph of the experimental setupFIG. 2
and -
FIG. 15 a photograph of a reaction vessel with laterally secured by a permanent magnet movable shutter.
In
Um die beiden Flüssigkeiten zu vermischen kann nun das Reaktionsgefäß 1 bewegt werden. Alternativ kann auch durch ein magnetisches Wechselfeld der bewegliche Verschluss 5 in Bewegung versetzt werden (
Das in
Die Nanopartikel können über an sie konjugierte Nukleinsäuren spezifisch an die durch PCR vervielfachten Nukleinsäuren binden, wodurch eine Änderung des Absorptionsspektrums der Flüssigkeit erzeugt werden kann. Vor der Messung der Absorption wird der bewegliche Verschluss 5 durch das Anlegen eines inhomogenen magnetischen Feldes aus der Flüssigkeit entfernt und im oberen Bereich des Reaktionsgefäßes 1 gehalten. Anschließend wird die Konzentration der gesuchten Nukleinsäuren durch Absorptionsmessung mittels eines Lasers 10 bestimmt.The nanoparticles can specifically bind to the nucleic acids amplified by PCR via nucleic acids conjugated to them, whereby a change in the absorption spectrum of the liquid can be produced. Before measuring the absorption, the movable shutter 5 is removed from the liquid by the application of an inhomogeneous magnetic field and held in the upper region of the
In einer weiteren Ausführungsform, wie sie in
In
In
mit einer äußeren ersten Unterteilung 6 und einer darin frei beweglichen zweiten
Unterteilung 7 in einem beweglichen hohlen Körper 20. Auch hier ist die zweite Unterteilung 7 zylindrisch aufgebaut und mit einem sphärischen, beweglichen Verschluss
5 verschlossen. Das Herausziehen des beweglichen Verschlusses 5 beim Einwirken eines inhomogenen magnetischen Wechselfelds basiert hier auf der niedrigeren Trägheit des beweglichen Verschlusses 5 im Vergleich zur übrigen zweiten Unterteilung 7. Dazu weist der bewegliche Verschluss 5 eine wesentlich geringere Masse als die zweite Unterteilung einschließlich der darin enthaltenen Flüssigkeit 7 auf.
with an outer
5 closed. The pulling out of the movable shutter 5 upon the action of an inhomogeneous alternating magnetic field is based here on the lower inertia of the movable shutter 5 in comparison to the remaining
Das in
In das Reaktionsgefäß 1 in
In
Das in
Sofern anschließend z.B. eine optische Transmissionsmessung erfolgen soll, kann die Kugel vorzugsweise durch einen kleinen Permanentmagneten 9 (z.B. 5mm Durchmesser und 10mm Länge) in einen Bereich des Reaktionsgefäßes verbracht werden, wo sie die weitere Untersuchung nicht beeinträchtigt. In
- 1. Reaktionsgefäß1st reaction vessel
- 2. äußere Öffnung des Reaktionsgefäßes2. outer opening of the reaction vessel
- 3. Deckel3. Lid
- 4. Schlauch4. hose
- 5. beweglicher Verschluss5. movable lock
- 6. erste Unterteilung6. first subdivision
- 7. zweite Unterteilung7. second subdivision
- 8. magnetische Komponente8. magnetic component
- 9. externer Permanentmagnet9. external permanent magnet
- 10. Laser zur Absorptionsmessung10. Laser for absorption measurement
- 11. Einzelmagnet11. Single magnet
- 12. Spalt zwischen zwei externen Permanentmagneten12. gap between two external permanent magnets
- 13. Kreiszylinder13. Circular cylinder
- 14. Kugel14. Ball
- 15. Aussparung in der ersten Unterteilung15. recess in the first subdivision
- 16. sich klammerartig an den beweglichen Verschluss anschmiegende W andabschnitte16 clinging to the movable closure clinging W
- 17. Boden der ersten Unterteilung17th floor of the first subdivision
- 18. oberer Wandbereich des Reaktionsgefäßes18. Upper wall area of the reaction vessel
- 19. Trennwand zwischen erster und zweiter Unterteilung19. Partition between first and second subdivision
- 20. beweglicher hohler Körper20. movable hollow body
- 21. Abstandshalter21. Spacers
- 22. konischer Anteil des Reaktionsgefäßes22. conical portion of the reaction vessel
- 23. elastischer Ring23. elastic ring
- 24. Folie24. Slide
- 25. zylindrischer Vorsprung des Deckels25 cylindrical projection of the lid
- 26. Elektromagnet26. Electromagnet
Claims (15)
- Reaction vessel (1) having a first subdivision (6) and a second subdivision (7), wherein in a first state of the reaction vessel (1) the first subdivision (6) is separated from the second subdivision (7) and in a second state the first subdivision (6) is connected to the second subdivision (7), wherein one of the two states is transferable into the other by the action of a magnetic field, and in the first state the first subdivision (6) and the second subdivision (7) are separated from one another by a movable closure (5) having a magnetic component (8), characterized in that in the first state the movable closure (5) is integrally connected to the reaction vessel (1).
- Reaction vessel (1) having a first subdivision (6) and a second subdivision (7), wherein in a first state of the reaction vessel (1) the first subdivision (6) is separated from the second subdivision (7) and in a second state the first subdivision (6) is connected to the second subdivision (7), wherein one of the two states is transferable into the other by the action of a magnetic field, and in the first state the first subdivision (6) and the second subdivision (7) are separated from one another by a movable closure (5) having a magnetic component (8), characterized in that at least one of the two subdivisions (6, 7) is located in a hollow body (20) movable in the reaction vessel (1).
- Reaction vessel (1) having a first subdivision (6) and a second subdivision (7), wherein in a first state of the reaction vessel (1) the first subdivision (6) is separated from the second subdivision (7) and in a second state the first subdivision (6) is connected to the second subdivision (7), wherein one of the two states is transferable into the other by the action of a magnetic field, characterized in that the first subdivision (6) is created on a cover of the reaction vessel by a film (24), and that a magnetic component (8) is located within the first subdivision (6).
- Reaction vessel (1) according to one of the preceding claims, characterized in that the magnetic component (8) is at least partially permanentmagnetic.
- Reaction vessel (1) according to one of the preceding claims, characterized in that in the first state the movable closure (5) is connected to the reaction vessel (1) in a non-positive manner.
- Reaction vessel (1) according to one of the preceding claims, characterized in that in the first state the movable closure (5) is positively connected to the reaction vessel (1).
- Reaction vessel (1) according to one of the preceding claims, characterized in that in the first state the movable closure (5) is materially connected to the reaction vessel (1).
- Reaction vessel (1) according to one of the preceding claims, characterized in that at least one of the subdivisions (6, 7) is at least partially limited by a cover (3) of the reaction vessel (1).
- Reaction vessel (1) according to one of the preceding claims, characterized in that the reaction vessel (1) is manufactured in one piece with at least one of the two subdivisions (6, 7).
- Reaction vessel (1) according to one of the preceding claims, characterized in that at least one of the two subdivisions (6, 7) is located in a hollow body (20) movable in the reaction vessel (1).
- Use of the reaction vessel (1) according to one of the preceding claims in a method for the purification, modification and/or detection of proteins and/or nucleic acids.
- Use of the reaction vessel (1) according to one of the preceding claims in a method for amplifying nucleic acids.
- Method for changing the state of the reaction vessel (1) according to one of claims 1 to 10.
- Method according to claim 13, characterized in that the change of state of the reaction vessel (1) is effected using a permanent magnet (9) or an electromagnet (26).
- Method according to one of the claims 13 or 14, characterized in that the change of state of the reaction vessel (1) is effected by using two permanent magnets (9) and/or electromagnets (26) which are arranged in such a way that there is a gap (12) between the permanent magnets (9) and/or electromagnets (26).
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102012222351.7A DE102012222351A1 (en) | 2012-12-05 | 2012-12-05 | Reaction vessel with magnetic closure |
PCT/EP2013/075637 WO2014086912A1 (en) | 2012-12-05 | 2013-12-05 | Reaction vessel having a magnetic closure |
Publications (2)
Publication Number | Publication Date |
---|---|
EP2928605A1 EP2928605A1 (en) | 2015-10-14 |
EP2928605B1 true EP2928605B1 (en) | 2019-09-04 |
Family
ID=49766052
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP13805819.3A Not-in-force EP2928605B1 (en) | 2012-12-05 | 2013-12-05 | Reaction vessel having a magnetic closure |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP2928605B1 (en) |
DE (1) | DE102012222351A1 (en) |
WO (1) | WO2014086912A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3984638A4 (en) * | 2019-06-13 | 2023-05-31 | Chin Hung Wang | Centrifugal reaction microtube, centrifugal reaction device and centrifugal test method therefor |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2000292325A (en) * | 1999-04-13 | 2000-10-20 | Shimadzu Corp | Head space gas sampler |
WO2006020182A2 (en) * | 2004-07-16 | 2006-02-23 | Smart Medical Technologies, Llc | Centrifuge system |
WO2006050636A1 (en) * | 2004-11-10 | 2006-05-18 | Capitalbio Corporation | A kind of net pcr reaction tube |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2086919A5 (en) * | 1970-04-14 | 1971-12-31 | Commissariat Energie Atomique | |
IT998660B (en) * | 1973-09-27 | 1976-02-20 | Erba Carlo Spa | ANALYTICAL CARTRIDGE CONTAINING THE SPECIFIC REAGENTS FOR DETERMINING THE SPECTROPHOTOMETRIC |
EP0515506B1 (en) | 1990-02-16 | 2000-01-05 | F. Hoffmann-La Roche Ag | Improvements in the specificity and convenience of the polymerase chain reaction |
US5576197A (en) | 1995-04-07 | 1996-11-19 | Molecular Bio-Products | Polymerase chain reaction container and methods of using the same |
AU6254796A (en) * | 1995-06-07 | 1996-12-30 | Becton Dickinson & Company | Device and method for phage-based antibiotic susceptibility testing |
US6153425A (en) | 1995-07-13 | 2000-11-28 | Xtrana, Inc. | Self-contained device integrating nucleic acid extraction, amplification and detection |
FI102642B (en) * | 1996-06-19 | 1999-01-15 | Orion Diagnostica Oy | Plug for a reaction vessel or equivalent |
DE602005000853D1 (en) * | 2004-07-02 | 2007-05-24 | Vivactis Nv | Measurement of heat generated by a chemical or biological process. |
US20060021673A1 (en) * | 2004-07-27 | 2006-02-02 | Stephan Rodewald | Self-sealing apparatus for chemical reaction vessel |
RU2009118455A (en) | 2006-10-17 | 2010-11-27 | Конинклейке Филипс Электроникс Н.В. (Nl) | DEVICE FOR AMPLIFICATION AND DETECTION OF NUCLEIC ACIDS |
KR101555476B1 (en) * | 2007-12-10 | 2015-10-06 | 삼성전자주식회사 | A thin film bio valve device and its controlling apparatus |
AT508708B1 (en) * | 2009-10-22 | 2011-06-15 | Gerhard Bonecker | TEST SET FOR A PHOTOMETRIC MEASURING DEVICE AND PHOTOMETRIC MEASURING METHOD FOR A SAMPLE LIQUID |
US20110239791A1 (en) * | 2010-01-29 | 2011-10-06 | Dolores Fici | System and method for biological sample storage and retrieval |
-
2012
- 2012-12-05 DE DE102012222351.7A patent/DE102012222351A1/en not_active Withdrawn
-
2013
- 2013-12-05 WO PCT/EP2013/075637 patent/WO2014086912A1/en active Application Filing
- 2013-12-05 EP EP13805819.3A patent/EP2928605B1/en not_active Not-in-force
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2000292325A (en) * | 1999-04-13 | 2000-10-20 | Shimadzu Corp | Head space gas sampler |
WO2006020182A2 (en) * | 2004-07-16 | 2006-02-23 | Smart Medical Technologies, Llc | Centrifuge system |
WO2006050636A1 (en) * | 2004-11-10 | 2006-05-18 | Capitalbio Corporation | A kind of net pcr reaction tube |
Also Published As
Publication number | Publication date |
---|---|
DE102012222351A1 (en) | 2014-06-05 |
EP2928605A1 (en) | 2015-10-14 |
WO2014086912A1 (en) | 2014-06-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE102014202838B3 (en) | Transport device, sample distribution system and laboratory automation system | |
EP2369343B1 (en) | Device and method for manipulating or examining a liquid sample | |
DE102009035941B4 (en) | diagnostic system | |
EP2030684B1 (en) | Device for providing pipettable substances | |
DE102005039175A1 (en) | Apparatus and method for separating magnetic particles from a liquid | |
DE10057396C1 (en) | Separation of e.g. biomolecules from dispersion or solution, employs magnetic particles onto which substance is sorbed, and electromagnet for their extraction | |
EP3025779B1 (en) | Incubation tray with test strip | |
DE112011103384T5 (en) | A method of observing a sample floating on a liquid surface in a scanning electron microscope | |
EP2599548B1 (en) | Sample holder for positioning an organic, biological and/or medical sample | |
JP2022136156A (en) | Microfluidic method for handling microdrops | |
EP3160646B1 (en) | Method and device for the transfer of liquids | |
EP3230730B1 (en) | Syringe housing for pipetting a biological material, comprising integrated membranes | |
EP2928605B1 (en) | Reaction vessel having a magnetic closure | |
DE102009005925B4 (en) | Apparatus and method for handling biomolecules | |
DE60007285T2 (en) | CAPILLARY LIQUID TRANSFER DEVICE INSIDE | |
WO2020094780A2 (en) | Device and method for transporting magnetic particles | |
EP3669981B1 (en) | Pressure-tight container containing a liquid | |
EP3573760A1 (en) | Carrier plate for laboratory devices | |
EP3034170A1 (en) | Supply holder for fluids | |
Ren | Magnetic Janus Particles and Their Applications | |
WO2019137775A1 (en) | Microfluidic device and method for the operation thereof | |
DE102013015522A1 (en) | Dialysis cell for in vitro release test apparatus, use of dialysis cell and in vitro release test apparatus | |
AT508394A1 (en) | DEVICE AND METHOD FOR THE APPLICATION OF RINGSTROMINDUCTION MODULATED BY MAGNETIC FIELD VARIATION IN ELECTRICALLY CONDUCTIVE NANO PARTICLES FOR MECHANICAL IMPACT ON CELLULAR MEMBRANES | |
DE202021105458U1 (en) | Device for the magnetic purification of biological samples | |
WO2023110944A1 (en) | Reaction-vessel unit, and methods for selectively removing a liquid from, and for introducing a liquid containing a target substance into, a reaction vessel of a reaction-vessel unit |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20150702 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
AX | Request for extension of the european patent |
Extension state: BA ME |
|
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20180220 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: GRANT OF PATENT IS INTENDED |
|
INTG | Intention to grant announced |
Effective date: 20190319 |
|
GRAS | Grant fee paid |
Free format text: ORIGINAL CODE: EPIDOSNIGR3 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE PATENT HAS BEEN GRANTED |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: FG4D Free format text: NOT ENGLISH |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: EP |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: REF Ref document number: 1174594 Country of ref document: AT Kind code of ref document: T Effective date: 20190915 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R096 Ref document number: 502013013535 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: FG4D Free format text: LANGUAGE OF EP DOCUMENT: GERMAN |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: MP Effective date: 20190904 |
|
REG | Reference to a national code |
Ref country code: LT Ref legal event code: MG4D |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: FI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: NO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191204 Ref country code: BG Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191204 Ref country code: SE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: HR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: LT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: RS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: AL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: ES Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: LV Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: GR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191205 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: PL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: PT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200106 Ref country code: NL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: IT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: RO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: EE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: SM Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: CZ Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200224 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R097 Ref document number: 502013013535 Country of ref document: DE |
|
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
PG2D | Information on lapse in contracting state deleted |
Ref country code: IS |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: DK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200105 |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
26N | No opposition filed |
Effective date: 20200605 |
|
REG | Reference to a national code |
Ref country code: BE Ref legal event code: MM Effective date: 20191231 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MC Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: SI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
GBPC | Gb: european patent ceased through non-payment of renewal fee |
Effective date: 20191205 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LU Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191205 Ref country code: GB Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191205 Ref country code: FR Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191231 Ref country code: IE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191205 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: BE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191231 Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191231 Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191231 |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: MM01 Ref document number: 1174594 Country of ref document: AT Kind code of ref document: T Effective date: 20191205 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: AT Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20191205 Ref country code: CY Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 Ref country code: HU Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO Effective date: 20131205 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: DE Payment date: 20211220 Year of fee payment: 9 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: TR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190904 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R119 Ref document number: 502013013535 Country of ref document: DE |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: DE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20230701 |