WO2009092571A2

WO2009092571A2 - Bubbles and method for producing and manipulating bubbles

Info

Publication number: WO2009092571A2
Application number: PCT/EP2009/000354
Authority: WO
Inventors: Thomas Franke; Lothar Schmid; Achim Wixforth
Original assignee: Universität Augsburg
Priority date: 2008-01-23
Filing date: 2009-01-21
Publication date: 2009-07-30
Also published as: DE102008005673A1; WO2009092571A3

Abstract

The invention relates to a bubble (2), particularly a vesicle or fluid cell, having a sheath (4), at least one liquid (6) trapped in the sheath (4), and at least one magnetic particle (8) trapped in the sheath (4), preferably a plurality of trapped magnetic particles (8). The invention also relates to a method for producing and manipulating the bubbles (2), and a device for this purpose.

Description

BUBBLES, AND METHOD FOR PRODUCING AND MANIPULATING BUBBLES

The invention relates to vesicles, in particular vesicles or fluid cells, to methods for the preparation and manipulation of the vesicles and to an arrangement for manipulating the vesicles.

Due to the rapid development in biotechnology, techniques are demanded with which the smallest amounts of liquid can be manipulated. The tendency towards miniaturization is driven above all by the following advantages: Small

Substances are cheaper and less toxic, often biological substances are available even in very small quantities. In addition, the reaction and analysis times are smaller by orders of magnitude and the specificity and efficiency is significantly higher. Therefore, microfluidic systems are already used today in so-called lab-on-a-chip systems, μ-TAS (total analysis system) and MEMS (micro-electro-mechanical system). Important components on such platforms are pumps, valves and integrated mixers. Such microfluidic systems also have disadvantages: the mixing is only diffusively possible due to the laminar flow. The transition from the laminar, hard-to-mix system to the turbulent system is characterized by the so-called Reynolds number, which relates inertial forces and viscous forces. If the Reynolds number is less than about 1000, this is referred to as the laminar regime in which adjacent layers of liquid flow "side-by-side" and can only diffuse diffusely, whereas if the Reynolds number is large, turbulences occur which dramatically accelerate intermixing

Liquid quantities are used eg surface wave mixing (SAW) (Wixforth A., Acoustically driven planar microfluidics, Superlattices Microstruct 2003; 33: 389-396). The principle here is to create specific vortexes in the liquid, which are much easier, virtually automatic, in macroscopic liquids, with the aim of creating the largest possible interface between the liquids to be mixed. Furthermore, the independent positioning of individual reagents is extremely difficult. However, controlled manipulation is of immense importance especially for applications. For example, in so-called "drug delivery" systems, various substances are included in vesicles (Szostak JW, Bartel DP, Luisi PL, Synthesizing life, Nature 2001, 409: 387-390). These are then injected, for example by means of syringes at the desired location in the body.

JP 2006045132 A discloses a liposome containing magnetic contrast agent for magnetic resonance measurements. US 5,078,986 A discloses a liposome-entrapped magnetic contrast agent which is released upon reaching a desired location in the body. US 4,728,575 A discloses vesicle-entrapped paramagnetic material as a contrast agent for NMR imaging. WO 2006/049497 A1 discloses permeable capsules which are produced by stabilization of vesicles. These capsules may contain magnetic particles as a diagnostic agent. The vesicles are generated by electrochemical polymerization. DD 290 807 A5 discloses vesicle-entrapped contrast agent for NMR. GB 2 078 660 A discloses magnetically-localizable, biodegradable lipid vesicles. The bubbles contain in addition to the magnetic material (magnetite) a medicinal substance. After injection of the bubbles into a body, the bubbles are held in place by a magnet (e.g., near a tumor) applied to the desired site to be treated. After a certain time, the shell of the bubble is broken down by the body (dissolved) and the active ingredient is released at the desired location.

It is an object of the invention to provide bubbles, bubbles in carrier liquid and a method for the production and manipulation of bubbles, can be manipulated as controllable as possible with the smallest amounts of liquid. This object is achieved by the features of claim 1, 15, 19, 35 and 39. Preferred embodiments are the subject of the dependent claims.

According to claim 19, a vesicle, in particular a vesicle or a liquid cell, is provided with a shell and at least one liquid enclosed in the shell, the shell including at least one magnetic particle. Preferably, a plurality of magnetic particles are included in the envelope. The at least one liquid may be a pure liquid or a liquid mixture. For example, a suspension, an emulsion or a solution. Through the envelope, the trapped liquid is shielded or separated, e.g. from a liquid surrounding the shell to prevent undesirable mixing with the environment.

A magnetic particle may be a single magnetic or consist of multiple components that together make up a magnetic particle.

Preferably, the envelope of the bubble is formed of polymers or comprises polymers. In this case, polymer molecules need not be present or not completely polymerized.

Alternatively or in an embodiment, the shell is made up of or essentially of amphiphilic molecules. In particular from lipids or amphiphilic polymers. That is, the starting material or "building blocks" of the shell have at least one polar and one nonpolar portion or one hydrophilic and one hydrophobic portion. Due to the attractive and repulsive forces of these building blocks, they assemble into self-assembled structures such as vesicles, (e.g., vesicles, cells) or membranes. For example, a shell of the lipids sphingomyelin, phosphatidylcholine and cholesterol can be constructed, which are also main components of red blood cells. Another starting material for a shell may be block polymers, i. Polymers, unlike

Homopolymers of at least two different monomer units A and B are constructed. Depending on the distribution of monomers A and B in the polymer chain A distinction is made between random (AABABBBAABB), alternating (ABABABAB) or block (AAAABBBB) copolymers, wherein at least one of the monomers is nonpolar and at least one other polar to form the structures described above.

Preferably, the shell has one or more layers. Especially with vesicles, a large number of different numbers of layers is possible. Preference is given to vesicles having a plurality of concentrically arranged layers. More preferably, the vesicles have a bilayer, i. they are unilamellar. Especially with the smallest trapped amounts of liquid, there is a single layer, i. a micelle.

Preferably, the envelope includes a volume on the order of up to 10 microliters, preferably up to 1 microliter, more preferably up to 100 femtoliters. This allows the smallest quantities of liquid to be separated from a surrounding medium by means of a bubble.

Preferably, the sheath comprises at least one functionalized building block, e.g. to form a channel in the shell. Preferably, the sheath is a membrane, in particular a liquid-permeable membrane, such as e.g. a cell wall. For example, the shell has special proteins embedded in the shell in order to let membrane-like only specific substances pass through the shell. The vesicle provides a depot for the specific substance, e.g. Medicament, ready, which can deliver a certain dose of this substance through the permeable shell. For example, a dose may be increased by increasing the number of channels so that more drug is released from within a bubble through the sheath to an environment.

Particular preference is given to using functionalized building blocks incorporated in the shell for marking the bubbles. For example, by incorporation of antibodies or adhesive molecules, bubbles can be bound to specific cells. When transporting the bubbles in a carrier liquid, these functionalized building blocks bind to the specific cells (with receptors for the functionalized building blocks) and an active ingredient contained in the vesicle can be deposited locally.

Preferably, the entrapped liquid is or contains an aqueous solution. For example, a sugar solution or an ionic solution, such as a

Saline. More preferably, the entrapped liquid is a suspension or an emulsion. For example, solids such as e.g. Medicines included. Additionally or alternatively, the entrapped liquid is an emulsion of a polar and a nonpolar liquid, such as e.g. Water and oil, o wherein preferably an emulsifier is added to the emulsion.

Preferably, at least two vesicles are provided, each of which includes different liquids or active ingredient components or reactants, in particular active substance components which react and / or are activated upon interaction

In a particularly preferred embodiment, at least one further vesicle is enclosed within the envelope, in particular at least one further vesicle. Preferably, several vesicles are included in the sheath. That is, within the bubble, which is a separate volume per se, another separated volume is arranged.

Preferably, at least in one of the further bubbles, a further liquid is enclosed, in particular a liquid which differs from the liquid surrounding the further bubble. This further liquid may contain an active substance 5 or an active ingredient component. By means of the further bubble or bubbles within the envelope, a plurality of active ingredient components or reactants can be deposited in the shell in a shielded manner from one another. In particular, in the case of active ingredient components which are activated by contact with one another, a vesicle is thus provided which activates active substances at a desired time and / or releases activated active substances, for example by destroying the shell of the active ingredient

Bubble and the envelope of the other bubbles at this desired time, whereby the active ingredients come into contact with each other. Preferably, the at least one magnetic particle is paramagnetic, in particular superparamagnetic. That is, when an external field is applied, magnetization of the particles occurs in the direction of the field. In addition, at least a part of the plurality of magnetic particles accumulates upon application of an external magnetic field to each other and in particular form at least one aggregate or a cluster within the shell. Particularly in the presence of an inhomogeneous magnetic field, the magnetic particles are drawn into regions with higher field strength, ie the particles follow a gradient of the inhomogeneous magnetic field. Furthermore, ferromagnetic particles can be used.

The at least one or the plurality of magnetic particles preferably have a diameter between 5 nanometers and 2 microns, preferably a maximum diameter of 10 microns, preferably of at most 1 micron or 50 nanometers. The magnetic particles may have any shape. Preferably, the magnetic particles are spherical or substantially spherical.

Particularly preferably, the magnetic particles are biocompatible. That when using the particles, these have e.g. in the human or animal body no harmful influence on this body and can e.g. be easily eliminated by this. Alternatively or additionally, the magnetic substance or elements of the particle are enclosed in a shell or matrix. For example, in latex, carbon or the like.

According to one embodiment, further particles are enclosed within the envelope of the bubble and the envelope forms a transport means for the further particles, in particular for drugs or drug components. Other particles include cancer markers, antibodies or vaccines.

According to claim 35, a combination of at least one carrier liquid and at least one bubble contained in the at least one carrier liquid is provided, wherein the at least one bubble be formed as described above can. Preferably, the vesicle contains an active ingredient which interacts with the carrier liquid, in particular reacts. Preferably, the active ingredient is activated by the interaction. An interaction can arise on the one hand by leakage of the active ingredient from the bubble through channels described above or by opening or at least partially destroying the envelope of the bubble.

The term "carrier liquid" is to be understood as the liquid carrying or containing the bubbles. This means that, on the one hand, a liquid for "storage" of bubbles - i. without this liquid contains components intended to interact with a drug of the bubbles - as

Carrier liquid is called. On the other hand, it is possible to add this carrier liquid with the bubbles, for example, to a blood sample, in which case the blood of the blood sample forms the carrier liquid for the bubbles.

The carrier liquid particularly preferably contains subunits with which an active substance contained in the vesicle or vesicles interacts, in particular reacts. For example, with cellular components such as blood cells when using blood as a carrier fluid.

More preferably, bubbles present in the carrier liquid in each case contain different active substances or components which interact or react with one another, in particular when interacting with one another.

The carrier liquid is preferably an aqueous solution or contains such, in particular one or more of the following: a sugar solution, ionic solution, blood, blood plasma.

According to claim 39, a method for producing at least one bubble, in particular a bubble as described above is provided. First, magnetic particles and at least one shell material are provided.

A starting material are, for example, amphiphilic polymers. At least one liquid is added to the magnetic particles and the starting material. The cover of a The forming bubble includes at least a portion of the liquid and at least one of the magnetic particles. After the formation, there is thus a volume separated from the surrounding liquid.

The at least one starting material preferably has at least one polymer former. In particular, the polymers described above can be used.

Particularly preferably, after the addition of the at least one liquid to the starting material, an electrical voltage, in particular an alternating voltage, is applied in order to promote the formation of bubbles.

In a particularly preferred embodiment, the magnetic particles and the at least one starting material are provided by applying a plurality of magnetic particles to a carrier substrate and a solvent containing the starting material to the carrier substrate. After stripping off, evaporating off and / or allowing the solvent to evaporate, the starting material for the at least one shell remains on the carrier substrate. The carrier substrate preferably has electrodes and / or the carrier substrate is an electrode in order to apply a voltage to the liquid as described above. The electrodes are preferably corrosion-free and / or biocompatible, in particular bioinert.

Preferably, electrodes are protected from corrosion by the application of a protective layer (e.g., quartz), the deposited layer forming the carrier substrate.

In order to assist in the formation of bubbles, in forming the bubbles, the temperature of the liquid is maintained in a range in which, in particular, the amphiphilic molecules are in a fluid phase. In the fluid phase, for example, a membrane formed from these molecules is "liquid", ie easily deformable. This ensures that, for example starting from a flat membrane surface, this membrane can curve or deform sufficiently to form a bubble. The added liquid preferably has further particles, such as functionalized particles and / or active substance substances, or further particles are applied to the carrier substrate. In this case, the further particles are enclosed in the bubbles during the formation of the bubbles and / or incorporated into the envelope. For example, the functionalized particles are cancer markers as described above.

According to claim 1, a method for manipulating at least one of a carrier liquid surrounded bubble is provided. Preferably, the vesicle is a vesicle as described above or prepared by the method described above. In particular, the vesicle has a shell with at least one liquid enclosed therein and at least one magnetic particle enclosed therein. The method provides for the application of an external magnetic field such that a force is applied to the at least one magnetic particle within the envelope of the bubble.

By applying a magnetic field, at least a portion of the plurality of magnetic particles preferably accumulates on one another and, in particular, forms a multiplicity of aggregates or clusters, in particular of oblong shape.

Preferably, the magnetic field is temporally changed and additionally or alternatively spatially changed, whereby the at least one magnetic particle or an aggregate of magnetic particles is influenced, in particular is moved. Preferably, the magnetic field has a spatial gradient and / or a magnetic field density change in order to exert spatially or temporally variable forces on the magnetic particle or particles.

More preferably, the magnetic field is a rotating magnetic field with which a magnetic particle is a cluster of magnetic particles is excited to rotate. Such a rotation of the magnetic particles or aggregates can be generated by spatially rotating permanent magnets or coils. Another

Possibility is arranging several coils next to each other, through alternating or successive switching of the coils is also a moving (eg a rotating magnetic field) is generated.

In particular, by applying a magnetic field, the at least one magnetic particle is moved within the envelope or moved relative to the carrier liquid. This results in the possibility of moving a bubble by means of the magnetic particles contained within the bubble. In particular, due to the force transmitted to the magnetic particles, a bubble can be moved independently of or even against flows outside the bubble. For example, a vesicle can be transported in this way in the bloodstream, regardless of the blood flow to a specific location or placed there or detained. Furthermore, for example, in this way a bubble can be positioned or transported on a fluidic analysis chip independently of a flow direction on the chip.

The movement of the particle through a magnetic field is preferably a linear movement, a rotational or circular path movement or a combination of linear and circular path movement. That The magnetic particle can be moved by a magnetic field on a predetermined or predetermined path. In particular, e.g. By stimulating rotation and / or vibration of the magnetic particle, mixing of a plurality of liquids present within the envelope can be improved or accelerated.

When providing at least one further bubble within the envelope, the envelope of the one further bubble can be destroyed or at least partially destroyed by a movement of the magnetic particles, for example by their rotation. Thus, for example, a liquid or an active ingredient of this further bubble can be released. In turn, this agent can then react with or activate the fluid within the sheath. This is particularly advantageous if an active ingredient to be activated is effective only for a short time after activation. Thus, in the vesicles smallest amounts of different drug or Reaction components are stored to be brought to react only when needed.

Preferably, the envelope of the bubble and / or the further envelope of the additional bubble (s) may be at least partially destroyed in order to contain the bubble contained therein

Liquids and / or agents, e.g. the above-described activated agents to release into the carrier liquid. Preferably, the shell is at least partially destroyed by means of at least one magnetic pulse. Further possibilities for destruction or opening of the envelope (envelopes) are the application of an electric field or ultrasound and additionally or alternatively microwave radiation. The energy of microwave rays is mainly absorbed by the magnetic particles and, in particular, has little influence on an aqueous carrier liquid. Alternatively, a solvent may be added to the carrier liquid which dissolves the shell of the bubble (s).

Preferably, the liquid released after the opening or at least partial destruction of the shell is mixed with the carrier liquid by means of the likewise released magnetic particles as described above, by applying a magnetic field or the magnetic field remains applied. In particular, a rotation of the released magnetic particles is excited. Thus, mixing of the released liquid with the carrier liquid is improved or accelerated and an efficient and rapid reaction or activation of a released active substance is made possible, in particular at the place of release.

Particularly preferably, the liberated liquid reacts with subunits contained in the carrier liquid. For example, when using blood as a carrier fluid, the released fluid reacts with cellular blood components, such as platelets. This reaction is also improved or accelerated by the above-described mixing by means of the magnetic particles. In particular, by the method, a liquid contained in the bubbles specifically in contact with, for example, certain organs or vessel walls (eg, veins) be brought and thus improve or accelerate a desired reaction of the released liquid with these.

In a further preferred embodiment, first bubbles with at least one first active component and second bubbles with at least one second

Active ingredient component provided in a carrier liquid. The at least one first active ingredient component is intended to react with the at least one second active ingredient component. For example, the vesicles with the respective active ingredient components can be prepared separately and then combined to provide a carrier liquid with vesicles each containing different active ingredient components. The active substances in the bubbles can be released in the manner described above, in particular after a positioning or transport of the bubbles, wherein a reaction of the first with the second active component by the method described above or mixing by means of the magnetic particles is improved.

By means of the method described, it is thus possible to specifically position an active substance or a reagent and to selectively activate and release it at this position. In addition, mixing with a carrier liquid, with subunits of a carrier liquid and / or a mixture of different ones is released

Drug components improved. This method can be used for example in the body (in vivo), but also in the laboratory (in vitro), for example on so-called analysis chips for DNA, PCR or blood analysis. Another area of application is the exact positioning of reactants in the synthesis of nano-structured substances.

Preferably, after release or after mixing by applying a magnetic field, the at least one magnetic particle or the plurality of magnetic particles is removed from the carrier liquid or locally concentrated in the carrier liquid. Especially when using this method in fluidic

Analysis devices so the carrier liquid can be replaced or removed without removing the magnetic particles. Ie after the exchange of a Carrier liquid, the same magnetic particles are available for further experiment or test.

According to claim 15, an arrangement for manipulating bubbles is provided with a carrier liquid and a bubble contained in the carrier liquid, the bubble having a shell and at least one magnetic particle contained in the shell. Furthermore, at least one magnetic field generator is provided which applies a magnetic field to the bubble, in particular a spatially and additionally or alternatively temporally variable magnetic field. In particular, with such an arrangement, a bubble as described above can be manipulated according to the methods described above.

Preferably, the at least one magnetic field generator is suitable for generating a focused magnetic field, wherein the at least one magnetic field generator has a device for selectively positioning a generated magnetic field.

When used in the laboratory, a device for receiving the carrier liquid with the bubbles contained therein is preferably arranged separately from the magnetic field generator, so that the carrier liquid can easily be exchanged for various experiments. Alternatively, the receiving device is designed as a disposable product, so that after an experiment or a test no cleaning of fluidic components is necessary.

In summary, by means of the described method, apparatus and vesicles, the smallest amounts of liquids and / or active substances-separated from a surrounding medium-can be purposefully transported and positioned, and the liquids / active substances can be deliberately released at this position. Additionally, prior to release of the fluids / agents or after release, mixing thereof within the vesicle or with a carrier fluid is enhanced. With reference to figures exemplary embodiments of the invention are explained in detail. Show it:

1 a source chamber for generating bubbles, FIG. 2 a first exemplary embodiment of a bubble, FIG.

3 shows a further embodiment of a bubble, in which a further bubble is contained,

FIG. 4 shows the bubble of FIG. 2 under the influence of a magnetic field, FIG. 5 shows the bubble of FIG. 3 under the influence of a magnetic field, FIG. 6 shows the bubble of FIG

bubble,

FIG. 7 shows the bubble of FIG. 2 with the envelope open, FIG. 8 shows the bubble of FIG. 7 without influence of a magnetic field, and FIG. 9 shows a schematic representation of a device for manipulating bubbles.

FIG. 1 shows a source chamber 12 which is used for the production of vesicles 2. In the following, a method for producing vesicles 2 using an electric field is described by way of example. In this case, a thin lipid film is swollen in aqueous solution and the film is replaced by an alternating electric field and converted into vesicles 2.

In the examples described, two different lipids are used as the main constituent of the swelling solutions: DOPC (1,2-dioleyol-sn-glycero-3-phosphocholine) is a lipid that is accessible experimentally throughout

Temperature range in the fluid phase is. DMPC (1,2-dimyristol-sn-glycero-3-phosphocholine chloroform) can be converted to the gel phase at low temperatures. The first source solution is a mixture of 150 μl DOPC chloroform solution with a concentration of c = 2.5 mg / ml and 10 μl Texas Red ™ DHPE chloroform solution with a concentration c = 0.5 mmol / 1. The

The composition of the second source solution is the same, but the DOPC solution is replaced by a DMPC solution with the same concentration. When Carrier substrate 14, 14 'glass plates are used, which are preferably coated with indium tin oxide (ITO). Since the plates are transparent, the formation of the vesicles 2 in the source chamber 12 can be observed under the microscope. The conductive ITO coating serves as an electrode.

As magnetic particles Magnetbeads 8 are used. In order to be able to swell vesicle 2 with enclosed magnetic beads 8, before applying a lipid solution or swelling solution to the carrier substrate 14, 14 ', initially 10 μl of a magnetic bead solution is distributed onto the carrier substrate 14, 14' and dried in vacuo. Thereafter, 10 μl of the DOPC or DMPC source solution is applied to the carrier substrate 14, 14 '. The support substrate 14, 14 'is then placed in vacuum for 3 hours to allow the chloroform to evaporate. Then two carrier substrates 14, 14 'are glued together with a spacer 16, in particular a Teflon spacer, to the source chamber 12. The base area of the source chamber 12 is approximately 20 × 50 mm ² , the distance between the carrier substrates is approximately 2 mm. Subsequently, the source chamber 12 is filled with a liquid 6, in particular a sugar solution.

For swelling, a 10 Hz alternating voltage is applied to the carrier substrates 14, 14 'or to the electrodes by means of an alternating voltage source 18, which voltage is slowly increased from 0 to 2.6 volts. The actual source process takes about 18 hours.

As shown in Fig. 1, it can be observed under the microscope that initially form on the plates bubble-like membrane forms that slowly fill with liquid 6 and finally peel off, each forming a shell 4 of a vesicle 2. Some of the formed vesicles 2 contain single magnetic beads 8, for example up to thirty magnetic beads 8. For the most part the vesicles are unilamellar, ie they have two single-layered lipid layers (ie a lipid bilayer). The size of the vesicle 2 is between 10 and 60 microns, with some vesicles 2 can be up to 130 microns in size. After swelling, the vesicles 2 can be pipetted off and stored in the refrigerator. In the preparation of DMPC vesicles 2, it must be noted that the phase transition temperature of this lipid is 23 ° C. In comparison, the phase transition temperature of DOPC is 20 ° C. Since lipids must be in the fluid phase (L _α phase) during swelling to be sufficiently deformable, the source chamber 12 becomes 2 when producing DMPC vesicles incubated in a heat bath. Furthermore, during the transition from the fluid phase into the so-called gel phase, the area of a lipid bilayer is reduced by about 30%. In order to avoid a rupture of the vesicles 2 by the membrane contraction on cooling, the volume of the vesicles 2 must be previously reduced osmotically. In addition, sucrose is added to the DMPC-Vesicle solution to increase the total concentration to 300 mM. This increases the available volume in vesicles 2 and prevents vesicle 2 from bursting.

Fig. 2 shows an embodiment of a vesicle 2 made, for example, by the method described above, with a sheath 4 containing the liquid 6 and

Magnetbeads 8 includes. The shell 4 may comprise a plurality of layers.

Preferably, it is unilamellar, d. H. consists of a lipid bilayer. The

Liquid 6 may be a mixture of several liquids, or may additionally contain other functionalized particles (not shown), e.g. Active ingredients, o reagents or markers. The vesicle 2 forms a compartment shielded from the external environment or a container for the liquid 6 and / or the functionalized particles.

3 shows a further exemplary embodiment of a vesicle 2 'in its shell 4', a further vesicle 2 ", which is enclosed by beads 8 and a liquid 6 '.This further vesicle 2" contains a further liquid 6 " "may contain or be a substance suitable for reaction with the liquid 6 '. In particular, the liquids 6 ', 6 "are aqueous solutions, in particular biocompatible solutions, Preferably the liquids 6', 6" contain active substances 0 for medical or therapeutic treatment. For example, contain the

Fluids 6 ', 6 "vaccines or cancer markers Further vesicles 2 may be included in "further magnetic beads 8." The one or more further magnetic beads 8 'may also be provided instead of the magnetic beads 8.

Fig. 4 shows the vesicle of Fig. 1 in a carrier liquid 10 under the influence of a magnetic field M. By the application of a magnetic field M, the judge

Magnetbeads 8 after the magnetic field M and in particular form aggregates, wherein the beads are arranged in particular rod-shaped. When applying a spatially and / or temporally variable magnetic field, these units can be moved. In particular, these aggregates may be used as a kind of "micro stir bar" when rotated by rotation of the external magnetic field M. Thus, the liquid 6 can be mixed in the interior of the shell 4 by means of the aggregates or the Magnetbeads 8.

This is particularly advantageous when there is another vesicle 2 "in the sheath 6 'of the vesicle 2' as shown in Fig. 2. As shown in Fig. 5, by the movement of the magnetic beads 8 by means of the external magnetic field M, the sheath 4 "further vesicle 2" are at least partially destroyed or opened, so that the liquid 6 "contained in the interior of the second vesicle 2" is released .. Subsequently, by agitation (eg rotation) of the magnetic bead aggregates a mixing of the further liquid 6 " be improved or accelerated with the liquid 6 'of the vesicle 2'. Thus, at any given time, an active substance can be released from the further vesicle 2 "and activated in the targeted vesicle 2 ', without the thus activated liquid or the active ingredient being released into the surrounding carrier liquid 10.

FIG. 6 shows how by applying a magnetic field, for example, the vesicle 2 of FIG. 1 can be moved in a carrier liquid 10. As a result of the fact that the aggregates or even individual magnetic beads 8 experience a force in the magnetic field M-in this representation the force action is directed to the right-they move inside the vesicle 2 or relative to the carrier liquid 10. In particular, a magnetic bead

8 or the Magnetbeads 8 pulled into a focus of the magnetic field M or follow the Focus of the magnetic field M. Thus, the vesicle 2 can be selectively transported and positioned by the force exerted on the beads 8 force.

FIG. 7 shows the release of the liquid 6 of the vesicle 2 from FIG. 1. For example, the magnetic beads 8 are accelerated in a jerky manner by means of a magnetic pulse and pass through the sheath 4 of the vesicle 2. Another possibility for opening the sheath 4 results a movement of the beads 8 at a speed which the vesicle 2, for example due to the viscous counter forces of the carrier liquid 10 can not follow. In this case, a part of the shell 4 is deformed by the beads or pulled in the length until the shell 4 breaks due to the deformation. By opening or at least partially destroying the shell 4, the liquid 6, as well as the beads 8 are released into the carrier liquid 10. In the same way, an activated active substance as described above is released.

After the release, as described above, by means of movement of the - now in the

Carrier liquid 10 present - Magnetbeads 8 mixing or mixing of the liquid 6 with the carrier liquid 10 are accelerated.

Fig. 8 shows the opened vesicle 2 of Fig. 7 without applied magnetic field M. The aggregates separate and there are again single magnetic beads 8 before. The

Magnetic beads can be separated from the carrier liquid 10 again by means of a magnetic field M or be separated from it. When using the vesicles in the body (for example of vesicles 2, 2 'in the blood as the carrier liquid 10) biocompatible Magnetbeads 8 are used, so that the beads 8 have no harmful effect on the body and, for example, can be easily eliminated from the body. In particular, the magnetic beads 8 have a biocompatible coating, such as a latex sheath.

9 shows a schematic representation of a device 20 for the manipulation of vesicles 2, 2 '. By means of permanent magnets 22, 22 'attached to a rotor 24, a rotating magnetic field is generated below a carrier liquid 10 arranged on a carrier 21. As a result, vesicles 2 contained in the carrier liquid 10, 2 'manipulated with Magnetbeads 8 in the manner described above. In particular, a liquid 6, 6 'contained in the vesicles 2, 2' and / or an active ingredient is released and mixed with the carrier liquid 10.

LIST OF REFERENCE NUMBERS

2, 2 'vesicles

4, 4 ', 4 "shell

6, 6 ', 6 "liquid

8 magnetic beads

10 carrier liquid

12 source chamber

14, 14 'carrier substrate

16 spacers

18 alternating voltage source

20 device

21 carriers

22, 22 'permanent magnet 24 rotor

Claims

claims:

Method for manipulating at least one vesicle (2, T) surrounding a carrier liquid (10), in particular a vesicle according to one of Claims 19 to 38 or a vesicle produced by the method according to one of Claims 39 to 46, the vesicle ( 2, 2 ') has a shell (4, 4') with at least one liquid (6, 6 ') enclosed therein and at least one magnetic particle (8) enclosed therein, and has the method:

Applying an external magnetic field (M), so that a force is exerted on the at least one magnetic particle (8).

2. The method of claim 2, wherein the carrier liquid (10) is an aqueous solution or contains, in particular is a body fluid, in particular blood.

3. The method of claim 1 or 2, wherein at least a part of the plurality of magnetic particles (8) upon application of a magnetic field (M) attach to each other, in particular form a rod, a needle, an aggregate and / or a cluster.

4. The method of claim 1, 2 or 3, wherein the magnetic field (M) changes temporally and / or spatially.

5. The method according to any one of claims 1 to 4, wherein the magnetic field (M) has a spatial gradient and / or a magnetic field density change, in particular, the gradient is spatially and / or temporally changed.

6. The method according to any one of claims 1 to 5, wherein the magnetic field (M) is a rotating magnetic field, in particular by a rotating magnetic field generator is generated, in particular by at least one permanent magnet (22) or at least one coil.

7. The method according to any one of claims 1 to 6, wherein the at least one particle (8) within the shell (4, 4 ') and / or relative to the carrier liquid (10) is moved by the magnetic field (M).

8. The method of claim 7, wherein the movement is a linear movement, a rotational or circular path movement or a combination of linear and circular path movement.

9. The method of claim 7 or 8, wherein by the movement of the at least one magnetic particle (8), the liquid (6, 6 ') in the shell (4, 4') and / or the carrier liquid (10) stirred and / or is mixed, in particular after an aggregation of a plurality of magnetic particles (8).

10. The method of claim 7, 8 or 9, wherein by the movement of the at least one magnetic particle (8), the sheath (4, 4 ') of the bladder (2, 2') and / or a sheath (4 ") of an inside the bubble (2 ') contained further bubble (2 ") is at least partially destroyed.

11. The method according to any one of claims 1 to 10, wherein the sheath (4, 4 ', 4 ") by means of at least one magnetic pulse, an electric field, ultrasound and / or opened by means of microwave rays and / or at least partially destroyed.

12. The method according to any one of claims 1 to 11, wherein by applying or by applying the magnetic field (M) from the open and / or at least partially destroyed shell (4, 4 ') released liquid (6, 6') with the Carrier liquid (10) and / or subunits of the carrier liquid by means of at least one of the opened and / or at least partially destroyed shell (4, 4 ') released magnetic particle (8) is mixed.

13. The method according to any one of claims 1 to 12, wherein at least two bubbles contain mutually different liquids and / or active ingredients and by applying or by leaving the magnetic field (M) released from the open and / or at least partially destroyed shells, different liquids be mixed together by means of at least one released magnetic particles (8).

14. The method according to claim 12 or 13, wherein after the release and / or after mixing by applying a magnetic field (M), the at least one magnetic particle (8) and / or the plurality of magnetic particles (8) from the

Carrier liquid (10) removed and / or locally concentrated in the carrier liquid (10).

15. Arrangement for manipulating bubbles (2, T), in particular bubbles according to one of Claims 1 to 14 or 19 to 34, with a carrier liquid (10), a bubble (2, T) contained in the carrier liquid (10), wherein the bubble comprises a shell (4, 4 ') and at least one magnetic particle (8) contained in the shell, and at least one magnetic field generator for applying a magnetic field (M) to the bubble, in particular for applying a spatially and / or temporally variable magnetic field (M).

16. Arrangement according to claim 15, wherein the at least one magnetic field generator has at least one permanent magnet (22) and / or at least one coil.

17. Arrangement according to claim 15 or 16, wherein the at least one magnetic field generator is suitable for generating a focused magnetic field.

18. Arrangement according to claim 15, 16 or 17, wherein the at least one

Magnetic field generator means for spatially positioning a generated Magnetic field (M), in particular of the focused magnetic field and / or the spatially and / or temporally variable magnetic field.

19. vesicles (2, T), in particular vesicles or liquid cells, with a shell (4, 4 '), at least one in the shell (4, 4') enclosed liquid (6, 6 '), and at least one in the shell (4, 4 ') enclosed magnetic particles (8), preferably a plurality of enclosed magnetic particles (8).

20. Blister according to claim 19, wherein the sheath (4, 4 ') is formed from polymers or polymers.

21. A vesicle according to claim 19 or 20, wherein the shell (4, 4 ') is composed of or substantially of amphiphilic molecules, in particular of lipids or amphiphilic polymers.

22. A vesicle according to claim 19, 20 or 21, wherein the envelope (4, 4 ') comprises one or more layers.

23. A vesicle according to any one of claims 19 to 22, wherein the sheath (4, 4 ') a

Volume on the order of up to 10 microliters, preferably up to 1 microliter, up to 100 picoliters, 10 picoliters or 1 picoliter.

24. A vesicle according to any one of claims 19 to 23, wherein the sheath (4, 4 ') has at least one functionalized building block, in particular a channel-forming

Has shell module.

25. A vesicle according to any one of claims 19 to 24, wherein the sheath (4, 4 ') is a membrane, in particular a liquid-permeable membrane.

26. A vesicle according to any one of claims 19 to 25, wherein the trapped liquid (6, 6 ') is or contains an aqueous solution, a suspension and / or an emulsion, in particular a sugar solution and / or ionic solution.

27. A vesicle according to any one of claims 19 to 26, wherein within the shell (4 ') at least one further vesicle (2 "), in particular at least one further vesicle, is enclosed.

28. A vesicle according to claim 27, wherein at least in one of the further vesicles (2 ") a further liquid (6") and / or at least one magnetic particle (8) or further magnetic particles (8 ¹ ) is enclosed, in particular one of the the further bubbles (2 ") surrounding liquid (6 ') different liquid, an active ingredient and / or an active ingredient component.

29. A vesicle according to claim 28, wherein the further liquid (6 ") is an aqueous liquid.

30. A vesicle according to any one of claims 19 to 29, wherein the at least one magnetic particle (8) is paramagnetic, in particular superparamagnetic.

31. A vesicle according to any one of claims 19 to 30, wherein at least a portion of the plurality of magnetic particles (8) upon application or presence of an external field (M) attach to each other, in particular a rod, a needle, an aggregate and / or a Form clusters.

32. A vesicle according to any one of claims 19 to 31, wherein the at least one magnetic particle (8) has a diameter of between 5 nm and 2 μm, preferably a diameter between 10 nm and 1 μm, and / or a maximum diameter of 10 micrometers, preferably has a maximum of 2 microns, 500 nm, 100 nm or 50 nm.

33. A vesicle according to any one of claims 19 to 32, wherein the at least one magnetic particle (8) is biocompatible.

34. A vesicle according to any one of claims 19 to 33, wherein within the shell (4, 4 ') further particles are included, in particular functionalized particles and / or

Active substance substances, in particular medical or therapeutic substances.

35. Combination of at least one carrier liquid (10) and at least one vesicle (2, T) contained in the at least one carrier liquid (10) according to one of claims 19 to 34.

36. The combination as claimed in claim 35, wherein at least one active substance which interacts with the carrier liquid and / or with subunits contained in the carrier liquid, in particular reacts, is contained in the bubble or bubbles (2, 2 ').

37. Combination according to claim 35 or 36, wherein at least two bubbles contained in the carrier liquid each contain different components, in particular interacting with each other and / or reacting components.

38. A combination according to claim 35, 36 or 37, wherein the carrier liquid (10) is or contains an aqueous solution, in particular a sugar solution, ionic solution, blood and / or blood plasma.

39. A method for producing at least one bubble (2, T), in particular a bubble according to any one of the preceding claims, comprising the steps:

Providing magnetic particles (8) and at least one starting material for a shell, adding at least one liquid (6, 6 '), and Forming and / or forming at least one bubble (2, T), wherein the forming sheath (4, 4 ') includes at least a portion of the at least one liquid (6, 6') and at least one of the magnetic particles (8).

40. The method of claim 39, wherein the at least one starting material comprises at least one polymer former and / or amphiphilic molecules.

41. The method of claim 39 or 40, wherein after the addition of an electrical voltage, in particular an AC voltage is applied.

42. The method of claim 41, wherein after the application of the voltage, this is increased stepwise or continuously.

43. The method according to any one of claims 39 to 42, wherein the magnetic particles (8) and the at least one starting material is provided by:

Providing the magnetic particles (8) on a carrier substrate (14, 14 '), providing a solvent containing the at least one starting material on the carrier substrate (14, 14'), and

Stripping and / or evaporation of the solvent.

44. The method according to any one of claims 39 to 43, wherein after adding, while forming and / or during the forming let a temperature of the liquid (6, 6 ') is maintained in a range in which the at least one polymer former and / or the amphiphilic molecules are in a fluid phase.

45. The method according to any one of claims 39 to 44, wherein the added liquid (6, 6 ') is or contains an aqueous solution.

46. The method according to any one of claims 39 to 45, wherein the added liquid (6, 6 ') further particles and / or further particles on the

Carrier substrate are provided, in particular functionalized particles and / or drug substances, in particular medical or therapeutic substances. 3.1

Fig. 1

2.3

3.3

2