WO2005002729A1

WO2005002729A1 - Use of a disposable container, microfluidic device and method for processing molecules

Info

Publication number: WO2005002729A1
Application number: PCT/EP2003/007645
Authority: WO
Inventors: Jürgen SCHÜLEIN; Björn GRASSL; Jürgen KRAUSE; Roland Barten; Dirk Kuhlmeier; Jörg HASSMANN; Hans Kosak
Original assignee: november Aktiengesellschaft Gesellschaft für Molekulare Medizin
Priority date: 2003-07-04
Filing date: 2003-07-15
Publication date: 2005-01-13
Also published as: EP1641564A1; ATE376882T1; DE50308515D1; EP1641564B1; AU2003250952A1

Abstract

The invention relates to the use of a disposable container comprising a cylinder (1) containing a plunger (2, 24) that is displaceably guided in said cylinder and a connection (3) that is provided on a first end (E1) of the cylinder (1) lying opposite the plunger (2, 24), said container being designed to create a difference in pressure in a microfluidic device (13), to hold (6) an agent (F, P) for processing molecules and/or to act as a reaction vessel.

Description

Use of a disposable container, microfluidic device and method for processing molecules

The invention relates to the use of a disposable container, a microfluidic device and a method for processing molecules.

Automatic analyzers for carrying out chemical and biochemical reactions are generally known from the prior art. Solutions required for the reaction are removed from a storage container by means of a suction pump and fed to a sample room. In addition, automatic analysis devices are also known, in which the solutions required for the reaction are supplied, for example, by means of a piston pump. - In the known devices, the solutions can be contaminated disadvantageously. The required solutions are usually presented in a certain volume. In this respect, there is a risk of inaccurate filling by the user. Automatic analyzers must be carefully cleaned after each analysis. This is time consuming. Apart from this, even with careful cleaning, a residue can remain in the device. Such a backlog leads to falsification of results in later analyzes.

From the field of medicine, e.g. B. known from DE 33 90 336 Tl disposable syringes. Such disposable syringes are e.g. T. filled up. A piston guided in a cylinder is designed such that the liquid received in the cylinder can be pressed out manually by means of the piston. Ampoules for taking liquid medicines are also known from the medical field. Such drugs can be injected, for example, using a syringe. It is therefore not possible to fill an exact volume in the μl range.

So-called microfluidic devices for the detection of predetermined biochemical molecules are also known from the prior art. Such devices work with small volumes. This is a detection of biochemical molecules, for. B. of DNA possible. Such a device is known for example from EP 0 397 424 A2 or EP 0 189 316 B1.

The object of the invention is to eliminate the disadvantages of the prior art. In particular, a use, a microfluidic device and a method are to be specified which enable a simplified and as accurate as possible automated implementation of sample preparation for carrying out chemical detection reactions and / or chemical detection reactions. In particular, a simplified automated preparation of samples for the detection of biochemical molecules, such as DNA, is to be made possible.

This object is achieved by the features of claims 1, 22 and

50 solved. Expedient configurations result from the features of claims 2 to 21, 23 to 49 and 51 to 59.

According to the invention, the use of a disposable container is provided, which has a cylinder with a piston displaceably guided therein and a connection provided at a first end of the cylinder opposite the piston, for generating a pressure difference in a Microfluidic device and for providing a means for processing molecules and / or as a reaction vessel.

Such a disposable container can be manufactured inexpensively. It can be filled with a predetermined volume using suitable machines. The filling process can be carried out in such a way that contamination is excluded. The disposable container can not only serve as a storage container for the agent for processing molecules, but also as a reaction vessel. For this, e.g. B. first a means for processing molecules in a microfluidic device and then a solution in the disposable container are pressed or sucked out of the microfluidic device. The disposable containers are easy to handle. Operation of microfluidic devices is thus quick, simple and easy to carry out.

In the sense of the present invention, a “microfluidic device” is understood to mean a compact and handy device for carrying out an analysis, synthesis, purification, modification and / or increasing the concentration of molecules. Such a microfluidic device can form a closed system with respect to the environment. A sample taken up in the microfluidic device is moved to carry out the respective reaction via at least one channel provided therein and, if necessary, brought into contact with a liquid presented in the microfluidic device. The channel is designed in such a way that the liquid can be displaced exactly over a predetermined section and thus moved precisely to a predetermined location of the device. According to one embodiment, it is provided that the means for processing is a liquid, a gel or a solid or a combination thereof. The solid can comprise at least one of the following constituents: soluble or suspendable particles, lyophilisate, chromatographic material, preferably an ion exchanger or an affinity matrix. The processing agent can also be selected from the following group: lysis liquid, elution liquid, buffer solution, beads, enzymes, primers, reactants, reagents.

The proposed disposable containers are preferably kept ready as a kit. Such a kit can be pre-assembled for certain analysis or diagnostic purposes. It can contain, for example, solutions required for cell disruption as well as adsorbents for DNA. Advantageously, the disposable containers are designed differently depending on the type of agent for processing molecules presented therein, for example in their diameter or in their length. This can prevent a disposable container from being accidentally connected to an incorrect connection of a microfluidic system. In this case, a recording of a microfluidic device is carried out in such a way that only the correct disposable container can be used in it. Of course, it is also possible to avoid a mix-up or a corresponding print

To provide coloring on the disposable container.

The processing of the molecules can involve analysis, synthesis, purification and / or increasing the concentration of the molecules. Such reactions always take place in vitro. The disposable container is expediently completely filled with the processing agent. In this case, the means for processing is a liquid, a paste, in gel or the like. A complete filling is understood to mean an essentially bubble-free filling in which the proportion of gas bubbles is <5.0% by volume. is expediently <1.0% by volume, preferably <0.1% by volume, particularly preferably <0.01% by volume. Liquids are expediently degassed before the disposable container is filled. Filling is preferably carried out under sterile conditions. It is also possible to sterilize the disposable container after it has been filled. This increases the durability of the processing agent contained in the disposable container.

The piston can be made of an elastic material, preferably rubber or plastic. It can have at least one circumferential seal which is symmetrical in cross section. This enables the piston to be pushed back and forth repeatedly, with a complete sealing effect being guaranteed at all times.

According to a further embodiment, the piston is designed to correspond to the connection, so that a complete emptying of the cylinder and possibly the connection is possible when the piston rests on the first end. This enables a particularly precise procedure; there are no residual volumes left in the disposable container. If the connection has a volume, this volume is also emptied.

According to a further embodiment, the piston has a means for engaging a pushing and / or towing means. This enables the disposable container to be easily connected a device for moving the piston. The means for engagement can be a recess provided centrally in the piston, e.g. B. a hemispherical or conical recess to a thread, a bayonet or a snap lock or the like .. It may also be the case that a pushing and / or towing device is attached to the piston. It can be a rod or a cylinder. In this case, the pushing and / or towing means has a means for engaging in a pushing and / or towing device at the free end. The means for engagement can be an opening, radially protruding projections, a flange or the like. The means for engagement is expediently designed such that it can be connected to a push and / or towing rod of a push and / or towing device by inserting it.

According to a further embodiment, the cylinder is made of a transparent material. This allows a simple visual inspection. It can immediately be determined whether the cylinder is correct, e.g. H. is bubble-free, filled. The cylinder can also be made of an opaque material, in particular for receiving light-sensitive reagents. The cylinder is preferably made of a material that is inert for the means for processing contained therein.

The cylinder is expediently made of an elastic plastic, preferably polypropylene. Polyethylene or polycarbonate are also suitable for manufacturing the cylinder. A particularly good sealing effect is achieved when using an elastic material. According to a further embodiment, the connection is closed with a closure means. The closing means can be a rubber or plastic membrane, a ball, a cone or a closing cylinder. Advantageously, the ball, the cone and / or the locking cylinder are made of a plastic or glass which is inert to the received means for processing.

According to a further particularly advantageous embodiment, a radially inwardly projecting projection that blocks a displacement of the piston out of the cylinder is provided on a second end of the cylinder opposite the connection. This makes removal of the piston impossible. Unwanted manipulation of the disposable container is avoided. Furthermore, at the second end, a further projection projecting radially inwards and opposing a displacement of the piston in the direction of the connection can be provided. The further projection is expediently designed such that the piston can only be moved in the direction of the first end by applying a predetermined force. The force is expediently chosen such that the connection is brought into a correct connection position with the connection piece before the piston is displaced. As soon as this is the case, a further axial displacement of the disposable container is no longer possible. The force acting on the further projection is then so great that the resistance thus formed can be overcome and the piston can be displaced in the direction of the first end. The provision of the further projection, in addition to ensuring a proper and tight connection of the disposable container to the microfluidic device, has the further advantage that an undesired leakage of liquid received in the disposable container prior to the Position of a completely sealed connection is avoided. The piston can only be moved after a tight connection has been made. The further projection is expediently designed to be radially symmetrical. This prevents uneven wear or damage to the piston when the further projection is overcome.

According to a further embodiment, a means for automatically reading out information about the means for processing received in the disposable container is provided. The means can be a barcode, a transponder, a chip or a specific shape. A barcode can, for example, be printed on the outside of the cylinder or applied by means of a label. A transponder or a suitable chip can be cast in at a suitable location on the disposable container. Furthermore, it may be that the cylinder has a specific shape, for example projections or recesses provided on the outside, which contain the information in codified form.

According to a further provision of the invention, a microfluidic device for processing molecules is provided with a device having at least one channel for conveying a sample, at least two connecting pieces being provided on the channel for connecting two disposable containers, each of the disposable containers having a cylinder which is displaceably guided therein Piston and a connection provided at a first end of the cylinder opposite the piston, and wherein the disposable containers with the connection provided thereon can each be connected to one of the connection pieces, so that liquid can be conveyed through the channel by displacing one of the pistons. The proposed microfluidic device is particularly simple. The use of micro pumps and the like can be dispensed with. A pressure to move the liquid in the channel of the microfluidic device is generated by moving the pistons. The disposable containers not only serve to provide means for processing molecules, but can also serve as a reaction space. If at least two disposable containers are connected to the microfluidic device, it is possible, for example, to purify or disrupt biological material by moving a liquid back and forth between the disposable containers. The proposed microfluidic device is surprisingly simple. It is easy to handle. The use of the proposed disposable containers eliminates the need to manually supply exact volumes, for example of liquids required to carry out reactions. Contamination is practically impossible. With the proposed microfluidic devices, exact results can be achieved quickly and reliably.

According to one embodiment, the channel is a channel system composed of a plurality of channels connected to one another. The channel or the channel system can be meandering at least in sections. In connection with the channel, the device can have a microfluidic mixing chamber and / or a microfluidic reaction space and / or a microfluidic detection space and / or a bubble trap. The channel can have a diameter of at most 2 mm, preferably less than 1.5 mm. The device can also have at least one agent selected from the following group: sensor, electrode, temperature control unit, sieve, filter, membrane, affinity matrix, substance or magnet. Furthermore, a connecting channel connecting the connecting piece to the channel can be provided. In addition, an inlet opening connected to the channel, preferably closable by means of a first valve, can be provided. Furthermore, an outlet opening connected to the channel, preferably closable by means of a second valve, can be provided. The proposed designs of the facility enable a differentiated and automated process management. The arrangement and design of the channel are expediently chosen so that the desired reaction can be carried out simply and quickly.

Advantageously, liquid can be conveyed into the other disposable container by moving the pistons. The piston can be pushed back into other disposable containers. It may also be the case that the piston in the other container is pulled back by a tow bar.

In a particularly advantageous embodiment, each connection piece has a pipe socket, preferably a hollow needle, for opening a closure means closing the connection. The connector or the pipe socket can be closed with a further closure means. It can be a rubber or plastic branch or the like. This prevents contamination of the device and / or of a liquid placed therein. The disposable container can expediently be a disposable container with the features described above. Each of the disposable containers can have a connection corresponding to the connection pieces. This enables simple and tight connection of the disposable containers to the connector. The connection can be closed with a closure means, for example a plastic or rubber membrane, a metal foil or the like.

According to a particularly advantageous embodiment, the device has a means for fixing the disposable container in a position that is fixed relative to the connector. This prevents undesired loosening of the connection from the connection piece. Leakages cannot occur, in particular when a liquid moves from the disposable container into the device or from the device into the disposable container. The means for fixing can be, for example, a thread, a bayonet catch or the like. For example, the connector can have an internal thread and the connector an external thread. Contamination of the agent for processing the molecules contained in the disposable container is avoided.

Advantageously, the device has at least one, preferably cylindrical, recess corresponding to the outer diameter of the disposable container for guiding the connection of the disposable container into an engagement position with the connecting piece. This makes it easier to connect the disposable container. The proposed microfluidic device is compact, stable and robust. Due to its design, it can easily be inserted into a device for the automatic movement of the pistons. Another advantage of the compact design of the microfluidic device is that it can be transported, packaged and stored is particularly simple. A faulty, possibly insufficiently tight connection is prevented.

Furthermore, the means for fixing can have a means for holding the disposable container that is fully inserted into the recess and engages in the connector in a fixed position. This prevents unwanted loosening of the connector from the connector. The means for holding can be, for example, at least one first latching means which surrounds the second end of the disposable container.

According to a further embodiment, a distance between the first latching means and the connection piece is selected such that the disposable container can be inserted into the recess without opening a closure means provided thereon. This makes it possible to insert the disposable container into one of the predetermined recesses by the manufacturer. This prevents the disposable containers from being inserted incorrectly by the user. The device can be delivered pre-assembled with the disposable containers suitable for the respective purpose. Such a prefabricated device, which can be designed, for example, in the manner of a cassette, then only has to be inserted into a suitable device for the automatic movement of the pistons and filled with the sample to be processed. Incorrect operation and contamination are practically excluded with such a pre-assembled device.

The disposable container is expediently received in the recess with an unopened closure means. Second locking means can be provided on the cylinder of the disposable container and / or on the inner wall of the recess. The second rest means can be designed so that the disposable container, for. B. by pressure on the piston, is displaceable into a latching position in which the connector closes the connector liquid-tight. Such an embodiment is expedient if the connection piece or the pipe socket are not closed with a second connection means. The second locking means can also be designed such that the disposable container can be moved from the first to a second locking position, in which the connection closes the connection piece in a liquid-tight manner and the pipe socket pierces the closing means. In the second latching position, a connection is first established between the disposable container and the channel. It is then possible, for example, to push a liquid from the disposable container into the channel.

The channel is advantageously filled with liquid. This enables a bubble-free liquid column to be easily produced in the entire device. This enables the liquid to be moved particularly precisely in the channel.

According to a further embodiment, several recesses can be provided on one side of the device. This simplifies and reduces the cost of building a device for the automatic movement of the pistons of the disposable containers. In this case, such a device only has to be equipped with a plurality of push rods which are suitable for pressing down the pistons and which are moved according to a predetermined program. In addition, the parallel arrangement of the recesses contributes to a particularly compact embodiment of the device according to the invention. The device is advantageously made in one piece from plastic. The cost of manufacturing the proposed facility is not particularly high. The device can be manufactured, for example, by means of injection molding. The device combined from the device with the disposable containers can be manufactured overall without great effort. It can be kept ready in the manner of a disposable device.

According to a further embodiment, the device can have a means for automatically reading out information about means for processing included in the device. The means can be a barcode, a transponder, a chip or a specific shape of the device.

The microfluidic device can be designed as a kit that includes the device and several of the disposable containers according to the invention. The disposable containers can already be captively inserted into the recesses, the closure means of which are of course still closed. Such a kit or microfluidic device is particularly easy to store, transport and operate by the user.

According to another aspect of the invention, a method for processing molecules is provided with the following steps:

Provision of at least two disposable containers, each of which has a cylinder with one which is displaceably guided therein

Piston and has a connection provided on a first end of the cylinder opposite the piston, Providing a microfluidic device with at least one channel, at least two connecting pieces being provided on the channel for connecting the disposable containers,

Connecting the disposable containers with the connectors provided on the connectors,

Shift one of the pistons so that a liquid is conveyed in the channel.

The proposed method enables a particularly simple processing of molecules. The pressure for moving or conveying the liquid in the channel of the device is generated by moving the pistons in the disposable containers. In this way, the liquid can be easily transported from one place to another in the facility. In particular, no micropumps and the like are required. The exact positioning of the liquid in the device can reduce the number of valves required.

The liquid is expediently contained in one of the disposable containers. It is also possible that liquid is present in the channel. This enables the simple production of a bubble-free liquid column in the facility. The device can be kept as a disposable part. The liquid can be transferred from one container to the other. When filling one of the disposable containers, the piston accommodated therein is expediently displaced by the liquid pressure. In this case, it is not necessary to tow the piston by means of a special towing device in order to convey the liquid into the disposable container. By moving one of the pistons, a liquid in the channel can be preferably meandering section of the channel and / or a microfluidic mixing chamber and / or a microfluidic reaction space and / or a microfluidic detection space and / or a bubble trap. Moving the pistons enables an exact

Displacement of the liquid. The liquid can be processed according to a predetermined program at certain points on the channel. For this purpose, the predetermined section of the channel can have a meandering channel and / or microfluidic detection space. To control the movement of the liquid in the channel, at least one valve provided therein can also be opened and / or closed according to a predetermined program. The pistons also move according to the specified program, so that the liquid in the channel or in the disposable containers is moved according to specified reaction steps.

According to a further embodiment, the disposable containers are inserted into recesses provided on the device. This enables the connections provided on the disposable containers to be guided precisely to the connection pieces of the device. When connecting the connector to the connector, the disposable container is expediently fixed relative to the connector. For this purpose, a means for fixing can be provided. The fixation is expediently only when the connection is properly connected to the connector. In this way, the establishment of a leaky connection can be avoided.

According to a further embodiment, the disposable container can be moved into a first latching position in the recess provided on the device, so that the connection closes the connection piece in a liquid-tight manner. Furthermore, can the disposable container can be moved from the first into a second latching position, so that the connection closes the connector in a liquid-tight manner and the pipe socket pierces the closure means. In the second latching position, a connection is only established between the disposable container and the channel.

Advantageously, the liquid is conveyed in the device by moving the pistons in such a way that the steps required to carry out at least one of the following methods are carried out: washing, purification, PCR, detection. The aforementioned methods are particularly suitable for the detection of DNA from different samples.

Exemplary embodiments of the invention are explained in more detail below with reference to the drawings. Show it:

1 is a schematic sectional view of a first disposable container,

2 shows a schematic partial sectional view of a first microfluidic device,

3 shows the partial sectional view according to FIG. 2 with a disposable container inserted,

4 shows the partial sectional view according to FIG. 3 with a push rod,

5 shows the partial sectional view according to FIG. 4 with the piston partially pressed in, FIG. 6 is a schematic sectional view of a second disposable container,

7 shows a schematic sectional view of a microfluidic device with the second disposable container according to FIG. 6,

8 shows the arrangement according to FIG. 7, the second disposable container being connected to the microfluidic device,

9 shows the arrangement according to FIG. 8, further disposable containers being connected,

10 shows the arrangement according to FIG. 9, with a sample container being connected,

11 shows the arrangement according to FIG. 10, the sample container having been emptied,

12 shows the arrangement according to FIG. 11, wherein the second disposable container has been emptied,

13 shows a sectional view of a further microfluidic device and

14a-c sectional views of the connection and the connection piece in different locking positions.

Fig. 1 shows a schematic sectional view of a first

Disposable container. In a suitably made of a transparent plastic, such as polyethylene or polypropylene, cylinder 1 is slidably a, for. B. Piston 2 made of plastic or rubber slidably guided. A connection 3 is provided at a first end E1 of the cylinder 1 opposite the piston 2. The connection 3 is closed with a closure means 4, here for example in the form of a glass ball. A liquid F is taken up in the volume formed by the cylinder 1 and the piston 2 located therein. This can be, for example, a lysis liquid, an elution liquid, a buffer solution or the like. The piston 2 is formed on its side facing the first end E1 of the cylinder 1 corresponding to the first end E1 of the cylinder 1. In particular, it has a projection 5 corresponding to the shape of the connection 3. If the piston 2 is pressed completely to the first end E1, the projection 5 fulfills the connection 3, so that a complete emptying of the liquid F from the cylinder 1 and the volume formed by the connection 3 is possible. On the side of the piston 2 opposite the projection 5, a central recess 7, a central projection 5 or the like can be provided for the engagement of a push rod and / or towing bar. A second end of the cylinder 1 opposite the first end E1 is designated by the reference symbol E2.

FIG. 2 shows a sectional view of a microfluidic device 13, specifically a receptacle 6 for the disposable container shown in FIG. 1. The receptacle 6 has a cylindrical recess 7 corresponding to the diameter of the cylinder 1. The recess 7 is designed such that the disposable container can be inserted therein. The connection 3 is guided in such a way that it arrives correctly in an engagement position of a connection piece 8. The connecting piece 8 can comprise a hollow needle 9 with which the closure means 4 pierce or pressed into the cylinder 1 and thus a connection of the cylinder 1 with the microfluidic device 13 is achieved.

3 shows the disposable container connected to the connector 8 using the receptacle 6. To transfer the liquid F, the piston 2 can be pressed in the direction of the connecting piece 8 by means of a push rod 10. This situation is shown in Figs. 4 and 5.

Fig. 6 shows in cross section a second disposable container. At the second end E2, the cylinder 1 has a retaining means 11 pointing radially inwards. With the retaining means 11, a displacement of the piston 2 beyond the second end E2 is prevented. The piston 2 has on its side facing away from the connection 3 an engagement means 12 for connection 3 to a push rod and / or towing rod. As shown here, the engagement means 12 can be designed, for example, in the form of a stamp. Of course, other suitable configurations are also possible. In the present exemplary embodiment, the closure means 4 is designed as a membrane. It can be a plastic and rubber membrane. It is expedient to have a plastic film formed in one piece with the cylinder 1. The cylinder 1 can be produced together with the plastic film in one piece, for example by means of injection molding. As shown in FIG. 6, particles P can be suspended in the liquid F.

7 shows in cross section a microfluidic device 13 which has a plurality of further connecting pieces 14 lying next to one another. The further connecting pieces 14 are closed with a further closure means 15, so that one Contamination of the microfluidic device 13 is excluded. The further closure means 15 can also be a plastic film, a rubber membrane or the like. Each of the further connecting pieces 14 is connected via a connecting channel 16 to a channel 17 connecting the connecting channels 16. The channel 17 is also connected via a first valve 18 to an inlet opening 19 and via a second valve 20 to an outlet opening 21. The connecting channels 16 and the channel 17 expediently have a diameter in the range from 1 to 2 mm. They are incorporated in a base plate 22 made of transparent plastic. Each of the connecting pieces 8, 14 has a hollow needle 9 for piercing the closure means 4.

8 shows the microfluidic device 13, the second disposable container having the connection 3 being connected to the further connection piece 14. The hollow needle 9 penetrates the connection 3, so that a tight connection is established. The liquid F can now be moved through the hollow needle 9 into the connecting channel 16 and into the channel 17 by moving the piston 2 in the direction of the connection 3.

FIG. 9 shows a sectional view of the microfluidic device 13 according to FIG. 8, second disposable containers being connected to all further connection pieces 14 here. As can be seen from Fig. 9, the diameter of the second disposable container can be designed differently. If necessary, recesses 7 provided on the microfluidic device 13 can thus be made such that only certain disposable containers can be used therein. For example, incorrect operation can be caused by connecting a disposable container to a wrong further connector 14 can be prevented. The other disposable containers shown in FIG. 9 are otherwise identical to the second disposable container.

FIG. 10 shows the microfluidic device 13 according to FIG. 9, a syringe 23 in which sample liquid PF is received being connected to the inlet opening 19 here. The sample liquid PF can be pressed, for example, into the opposing second disposable container by means of the syringe 23. To do this, it is necessary to open the first valve 20 and to hold down another piston 24 of a further disposable container 25. In this case, the sample liquid PF is pressed via the connecting channel 16 into the disposable container opposite the syringe 23. A piston 24 located therein is displaced by the applied pressure (see FIG. 11).

The transferred sample liquid PF can then be mixed with the liquid F contained in the cylinder 1, for example by depressing the piston 2. For this purpose, it is necessary to depress the piston 2 and again to hold the second piston 24 in position, and to keep the first 18 and the second valve 20 closed (see FIG. 12). The first and / or second valve can be a simply designed one-way valve which has, for example, a spring-loaded ball or a spring-loaded cone as the valve body. Such a one-way valve is arranged such that a liquid flow in the direction of the channel is possible under pressure and the valve body is pressed into its closed position by a pressure generated in the channel. By moving the valve body away from its closed position, the valve can be emptied. The first and / or second valve, it can also be a simply designed reusable valve, e.g. B. a diaphragm valve act.

By means of a program-controlled predetermined movement of the pistons 2, 24 of the disposable containers, the sample liquid PF can be acted upon in succession with several different liquids. It can be mixed intensively with the liquids. For example, a simple disruption of cells contained in the sample liquid PF is possible. As soon as the cells are disrupted, DNA molecules contained therein can be separated by magnetic beads containing liquids in the disposable containers and can be sent to a further detection method for analysis.

FIG. 13 shows a further exemplary embodiment of a microfluidic device 13. In contrast to the microfluidic device 13 shown in FIGS. 7 to 12, it has cutouts 7 on one side, preferably in a parallel arrangement, for inserting disposable containers (not shown here) ) on. The recesses 7 have first latching means 26 at their ends opposite the connector 8. The first latching means 26 serve to ensure that a disposable container once inserted into the recess 7 can no longer be easily removed from the recess 7. On

Distance A between the first latching means 26 and the connection piece 8 is selected such that a disposable container can be pushed completely into the recess 7 without the hollow needle 9 piercing a closure means 4 provided at its connection 3. The disposable container can therefore be presented captively with the microfluidic device. To start up the device according to the invention, it is only necessary to remove the To move the disposable container in the direction of the connector 8 and thus to connect it to the microfluidic device 13.

The sectional views FIGS. 14a to 14c show an advantageous embodiment of the connection of the disposable container to the microfluidic device 13. Second latching means provided in the recess 7 in the vicinity of the connector 8 are designated by 27. In the area of the connection 3 of the one-way container, third locking means 28 are provided on the cylinder 1, which are designed to correspond to the second locking means 25. The hollow needle 9 is with a, for. B. executed as a 0-ring, sealant 29.

In the first position shown in FIG. 14 a, the disposable container is pushed into the recess 7 of the microfluidic device 13. There is no sealing connection between the connection 3 and the sealing means 29 provided on the hollow needle 9. The closure means 4 is closed.

In the second position shown in FIG. 14b, the disposable container is in a first latching position. In this position, the connection 3 overlaps the sealant 29. The connection 3 thus closes the opening formed by the hollow needle 9 in a liquid-tight manner.

14c, the disposable container is in a second latching position. The hollow needle 9 penetrates the locking device 4 (not shown here). There is a fluid connection between the hollow needle 9 and the disposable container. It is readily apparent from the exemplary embodiments that the proposed microfluidic device can be easily equipped with a wide variety of combinations of reagents, agents, buffers and the like. The proposed device can be used for a wide variety of assays. It is only necessary to fill the disposable containers with the necessary reagents, agents and the like. The disposable containers themselves and the microfluidic device do not have to be changed. Nevertheless, they can be made extremely inexpensively from injection molded plastic. This makes it possible to design the disposable containers as well as the microfluidic device as a disposable part. Nevertheless, extremely precise reaction management is achieved. Defined concentrations and volumes can easily be kept in the disposable containers. Since the proposed system is completely closed, contamination is avoided. The proposed device is outstandingly suitable for the automated detection of DNA or similar biochemical molecules. A device to be provided for automatic

Control of the movements of the pistons 2, 24 of the disposable containers can be carried out relatively easily and inexpensively. Such a device does not require a particularly high level of maintenance, since liquids do not escape from the proposed device. The proposed disposable containers allow simple and flexible storage, assembly and manufacture of the proposed device. LIST OF REFERENCE NUMBERS

1 cylinder

2 pistons

3 connection

4 fasteners

5 head start

6 recording

7 recess

8 connector

9 hollow needle

10 push rod

11 restraint devices

12 means of intervention

13 microfluidic device

14 additional connector

15 further closure means

16 connecting channel

17 channel

18 first valve

19 entrance opening

20 second valve

21 exit opening

22 base plate

23 syringe

24 more pistons

25 further disposable containers

26 first locking means

27 second locking means

28 third locking means

distance F liquid

P particles

PF sample liquid

Claims

claims

1. Use of a disposable container which has a cylinder (1) with a piston (2, 24) slidably guided therein and a connection (3) provided on a first end (E1) of the cylinder (1) opposite the piston (2, 24) has, for generating a pressure difference in a microfluidic device (13) and for providing a means (F, P) for processing molecules and / or as a reaction vessel.

2. Use according to claim 1, wherein the means (F, P) for processing is a liquid, a gel or a solid or a combination thereof.

3. Use according to claim 1 or 2, wherein the solid comprises at least one of the following constituents: soluble or suspendable particles, lyophilisate, chromatographic material, preferably an ion exchanger or an affinity atrix.

4. Use according to one of the preceding claims, wherein the agent (F, P) for processing is selected from the following group: lysis liquid, elution liquid, buffer solution, beads, enzymes, primers, reactants, agents.

5. Use according to one of the preceding claims, wherein the processing is an analysis, synthesis, purification, modification and / or increasing the concentration of the molecules.

6. Use according to one of the preceding claims, wherein the disposable container is completely filled with the processing means.

7. Use according to one of the preceding claims, wherein the piston (2, 24) is made of an elastic material, preferably of rubber or plastic.

8. Use according to one of the preceding claims, wherein the piston (2, 24) has at least one circumferential cross-sectionally symmetrical seal.

9. Use according to any one of the preceding claims, wherein the piston (2, 24) is designed to correspond to the connection (3), so that when the piston (2, 24) abuts the first end (El), the cylinder (1) is completely emptied. and possibly the connection (3) is possible.

10. Use according to one of the preceding claims, wherein the piston (2, 24) has a means (12) for engaging a pushing and / or towing means.

11. Use according to one of the preceding claims, wherein a pushing and / or towing means (10) is attached to the piston (2, 24).

12. Use according to one of the preceding claims, wherein the pushing and / or towing means (10) at the free end

Has means for engaging in a pushing and / or towing device.

13. Use according to one of the preceding claims, o- in which the cylinder (1) is made of a transparent or an opaque material.

14. Use according to one of the preceding claims, wherein the cylinder (1) is made of an elastic plastic, preferably polypropylene.

15. Use according to one of the preceding claims, wherein the connection (3) is closed with a closure means (4).

16. Use according to one of the preceding claims, o- in which the closure means (4) is a rubber or plastic membrane, a ball, a cone or a closure cylinder.

17. Use according to one of the preceding claims, wherein the ball, the cone and / or the locking cylinder are made of an inert plastic or of glass.

18. Use according to one of the preceding claims, wherein at a second end (E2) of the cylinder (1) opposite the connection (3) there is a radially inwardly projecting displacement of the piston (2, 24) out of the cylinder (1 ) blocking projection (11) is provided.

19. Use according to one of the preceding claims, wherein at the second end (E2) a radially inwardly projecting, a displacement of the piston (2, 24) in the direction of the connection (3) is provided a further projection.

20. Use according to one of the preceding claims, wherein a means for automatically reading out information about the means (F, P) accommodated in the disposable container is provided for processing.

21. Use according to one of the preceding claims, wherein the means is a barcode, a transponder, a chip or a specific shape.

22. Microfluidic device for processing molecules with a device (13) having at least one channel (16, 17) for conveying a sample (PF),

at least two connecting pieces (8, 14) are provided on the channel (16, 17) for connecting two disposable containers,

Each of the disposable containers has a cylinder (1) with a piston (2, 24) slidably guided therein and a connection (3) provided on a first end (E1) of the cylinder (1) opposite the piston (2, 24) , and

The disposable containers with the connection (3) provided thereon are each connected to one of the connection pieces (8, 14), so that liquid (F) is displaced through the channel (16, 17) by moving one of the pistons (2, 24). is transportable.

23. The apparatus of claim 22, wherein the channel (16, 17) is a channel system of several interconnected channels.

24. The device according to claim 22 or 23, wherein the channel (16, 17) or the channel system is at least partially changing-shaped.

25. Device according to one of claims 22 to 24, wherein the device (13) in connection with the channel (16, 17) a microfluidic mixing chamber and / or a microfluidic see reaction space and / or a microfluidic detection space and / or a bubble trap.

26. Device according to one of claims 22 to 25, wherein the channel (16, 17) has a diameter of at most 2 mm, preferably less than 1.5 mm

27. Device according to one of claims 22 to 26, wherein the device (13) has at least one selected from the following group: sensor, electrode, temperature control unit, sieve, filter, membrane, affinity matrix, substance, magnet.

28. Device according to one of claims 22 to 27, wherein a connecting channel (16) connecting the connecting piece (8, 14) to the channel (17) is provided.

29. Device according to one of claims 22 to 28, wherein an inlet opening (19) which is connected to the channel (17) and can preferably be closed by means of a first valve (18) is provided.

30. Device according to one of claims 22 to 29, wherein an outlet opening (21) which is connected to the channel (18) and can be closed, preferably by means of a second valve (20), is provided.

31. Device according to one of claims 22 to 30, wherein liquid (F) can be conveyed into the other disposable container by displacing one of the pistons (2, 24).

32. Device according to one of claims 22 to 31, wherein each connection piece (8, 14) a pipe socket, preferably has a hollow needle (9) for opening a closure means (4) closing the connection (3).

33. Device according to one of claims 22 to 32, wherein the connecting piece (8) and / or the pipe socket is closed with a further closure means (15).

34. Device according to one of claims 22 to 33, wherein the disposable container is a disposable container according to one of claims 1 to 20.

35. Device according to one of claims 22 to 34, wherein each of the disposable containers has a connection (3) corresponding to the connection pieces (8, 14).

36. Device according to one of claims 22 to 35, wherein the device (17) has at least one, preferably cylindrical, recess (7) corresponding to the outer diameter of the disposable container for guiding the connection (3) of the disposable container into an engagement position with the connecting piece ( 8, 14).

37. Device according to one of claims 22 to 36, wherein the device further comprises a means for fixing the disposable container in a fixed position relative to the connector.

38. Device according to one of claims 22 to 37, wherein the means for fixing has a means for holding the disposable container fully inserted into the recess (7) and engaging in the connection piece (8, 14) in a fixed position.

39. Device according to one of claims 22 to 38, wherein the means for holding is at least one first latching means (26) which surrounds the second end (E2) of the disposable container.

40. Device according to one of claims 22 to 39, wherein a distance (A) between the first latching means (26) and the connecting piece (8) is selected such that the disposable container can be inserted into the recess (7) without one provided closure means (4) is opened.

41. Device according to one of claims 22 to 40, wherein the one-way container (s) with unopened closure means (4) is / are received in the recess (7).

42. Device according to one of claims 22 to 41, wherein second locking means (27) are provided on the cylinder (1) of the disposable container and / or on the inner wall of the recess (7).

43. Device according to one of claims 22 to 42, wherein the second latching means (27) are designed such that the disposable container can be moved into a first latching position in which the connection (3) closes the connection piece (8) in a liquid-tight manner.

44. Device according to one of claims 22 to 43, wherein the second locking means (27) are designed such that the disposable container can be moved from the first into a second locking position, in which the connection (3) the connection piece (8) is liquid-tight closes and the pipe socket (9) pierces the closure means (4).

45. Device according to one of claims 22 to 44, wherein liquid is placed in the channel.

46. Device according to one of claims 22 to 45, wherein a plurality of recesses (7) are provided on one side of the device (13).

47. Device according to one of claims 22 to 46, wherein the device is made in one piece from plastic.

48. Device according to one of claims 22 to 47, wherein the device (13) has a means for automatically reading out information about means (F, P) recorded in the device (13) for processing.

49. The apparatus of claim 48, wherein the means is a barcode, a transponder, a chip or a specific shape.

50. Method for processing molecules with the following steps:

Providing at least two disposable containers, each of which has a cylinder (1) with a piston (2, 24) slidably guided therein and a connection (3) provided on a first end (E1) of the cylinder (1) opposite the piston (2, 24) ) having,

Providing a microfluidic device (13) with at least one channel (16, 17), at least two connecting pieces (8, 14) being provided on the channel (16, 17) for connecting the disposable containers, Connecting the disposable containers with the connectors (3) provided on them to the connectors (8, 14),

Shift one of the pistons (2, 24) so that a liquid (F) is conveyed in the channel (16, 17).

51. The method of claim 50, wherein the liquid (F) is contained in one of the disposable containers.

52. The method of claim 50 or 51, wherein the liquid (F) is conveyed from one to the other disposable container.

53. The method according to any one of claims 50 to 52, wherein when one of the disposable containers is filled, the piston (2, 24) received therein is displaced by the liquid pressure.

54. The method according to any one of claims 50 to 53, wherein by moving one of the pistons (2, 24) a liquid located in the channel (16, 17) into a predetermined, preferably meandering section of the channel (16, 17) and / or a microfluidic mixing chamber and / or a microfluidic reaction space and / or a microfluidic detection space and / or a bubble trap is shifted.

55. The method according to any one of claims 50 to 54, wherein to control the movement of the liquid in the channel (16, 17) at least one valve (18, 20) provided therein is opened and / or closed according to a predetermined program.

56. The method according to any one of claims 50 to 55, wherein the disposable container is inserted into a recess (7) provided on the microfluidic device.

57. The method according to any one of claims 50 to 56, wherein the disposable container in the recess (7) is moved into a first latching position, so that the connector (3) closes the connector (8) in a liquid-tight manner.

58. The method according to any one of claims 50 to 57, wherein the disposable container is moved from the first into a second latching position, so that the connection (3) closes the connection piece (8) in a liquid-tight manner and the pipe socket (9) pierces the closure means (4) ,

59. The method according to any one of claims 50 to 58, wherein the liquid (F) is conveyed in the device (13) by displacing the pistons (2, 24) in such a way that the steps required to carry out at least one of the following methods be carried out: washing, purification, PCR, detection.