DE102005000834B4 - Method and device for metering and mixing small quantities of liquid - Google Patents

Abstract

The invention relates to a method for the integrated metering and mixing of small amounts of liquid with a volume of up to 1 ml, in which at least one metering reservoir is completely filled with a first liquid which communicates with a reaction reservoir via at least one connecting structure, wherein the at least one Connection structure is designed such that the entry of the first liquid is initially prevented in the reaction reservoir, and the reaction reservoir is completely filled with a second liquid, so that the second liquid at the connection structure with the first liquid in communication. In the liquid at least in or on the reaction reservoir, a flow pattern for mixing the liquids is generated. The invention further relates to a device and an apparatus for carrying out the method according to the invention and a use.

Description

The The invention relates to a method for metering and mixing small amounts of liquid, an apparatus and an apparatus for carrying out the method and a Use.

diagnostic Assays, in particular in the field of clinical chemistry and immunochemistry become big these days Part done automatically. In the corresponding machines are defined volumes of sample liquid and reagents in a cuvette or pipetted into the well of a microtiter plate and mixed. Subsequently a first reference measurement is carried out, in which, for example, the optical transmission through the cuvette is determined. After a certain reaction time between sample and reagents becomes a second Measurement of the same parameter made. By comparison The two measured values result in the concentration of the sample with respect to one certain ingredient or just the presence of the ingredient. Typical volumes are a few hundred microliters in total, where necessary mixing ratios from sample to reagent between 1: 100 and 100: 1. Possibly can also provided several reagents for mixing with a sample be. In addition to the instruments just described for high Throughput, typically found in specialized laboratories, There are also efforts, assays decentralized and without major instrumental To carry out effort. It would be desirable, if the most recent Time introduced "lab-on-a-chip" technology could be in the processing of liquids can be integrated on or integrated in a chip. assay times less than an hour are desirable.

to Movement of liquids For example, microfluidic systems are used here, in which liquid is moved by electro-osmotic potentials, see for example Anne Y. Fu, et al. "A micro fabricated fluorescence-activated cell sorter ", Nature Biotechnology, Vol. 17, November 1999, p. 1109-1111

A method for mixing liquids in the microliter range is in DE 103 25 307 B3 described in which small volumes of liquid in microtiter plates are mixed by means of sound-induced flow. Another method of generating motion in small quantities of fluid on a solid surface describes DE 101 42 789 C1 , Here, a liquid is mixed with the help of surface sound waves or mixed several liquids together.

According to a in DE 100 55 318 A1 described method, an amount of liquid is brought to a region of a substantially planar surface, the wetting properties of which differ from the surrounding surface in such a way that the liquid preferably resides on it, being held together by their surface tension. Movement of the amount of liquid can be generated by the momentum transfer of a surface acoustic wave to the liquid.

Problematic is in particular the integration of dosage and mixture of Sample and reagents in a low-cost lab-on-the-chip system. A homogeneous mixing of different such small amounts of liquid is difficult to realize.

For dosing, it is necessary to precisely define volumes of liquid quantities. This is geometrically feasible, for example. For example, in an open system, the wetting properties of the surface can determine a volume, as in FIG DE 100 55 318 A1 is described. Here, the volumes are defined by hydrophilic and hydrophobic regions over the wetting angle on a substantially smooth surface. If several volumes have been defined in this way which are to be reacted, the volumes are moved toward one another in order to achieve this. When moving on a surface, liquid residues or liquid molecules of the analyte or the reagent can adhere to the surface, so that the movement of a volume loss or a concentration reduction of unknown level can not be excluded. In addition, provision must be made against the evaporation, which can be problematic especially at longer assay times.

Other approaches use channels of defined cross-section, which are filled with liquid capillary. If the liquid is an aqueous solution, then at the end of the channel a hydrophobic barrier is attached, which can not be filled capillary. Furthermore, there is a lateral branch on this channel with a likewise hydrophobic surface, which can not be capillary filled. The cross-section and length of the channel between the hydrophobic barrier and the hydrophobic branch now determine a volume which can be separated and moved by pneumatic pressure through the branch (Burns et al., An integrated nanoliter DNA analysis device, Science, Vol. 282, 1998, pp. 448-487). This type of volume definition creates high costs due to the necessary wetting structure of the surface (hydrophilic for filling the channel itself and hydrophobic for the barrier and the Ab branch). In addition, you must work with air pressure, which requires appropriate devices. In order to enable the capillary filling of the measuring channel, the channel cross-section must be small. For large volumes in the range of a few 100 microliters, therefore, long channels are required. This inevitably leads to large unwanted interactions of the molecules in the liquid with the channel wall. An efficient mixing of several quantities of liquid is almost impossible in this geometry.

Of the The term "liquid" as used herein Text including pure liquids, Mixtures, dispersions and suspensions and liquids, in which solid particles, for example biological material, are located. To be dosed and mixed liquids can for Example also be two or more solutions, which only through dissolved in it Differentiate ingredients that are to be reacted.

task It is the object of the present invention to provide a method and an apparatus with the help of which a precise dosage of liquid quantities in a big one dynamic range is easy to perform and which requires complete mixing of liquids enable. The method should be feasible in a compact lab-on-the-chip system.

These The object is achieved by a method having the features of the claim 1, a device with the features of claim 17 or an apparatus solved with the features of claim 35. Subclaims are directed to preferred embodiments. An advantageous use is the subject of claim 41.

at the method according to the invention For integrated dosing and mixing, a dosing reservoir is used with a first liquid Completely filled, the above at least one connecting structure with a reaction reservoir is in communication, wherein the connection structure is preferably such in relation to dimensioned to the reservoirs that the surface tension the first liquid prevents entry into the reaction reservoir. Especially For this purpose, the cross section of the connection structure can be chosen to be smaller as the cross section of the reaction reservoir. The reaction reservoir comes with a second liquid Completely filled, leaving the second fluid at the connection structure comes into contact with the first liquid. After all gets in the liquid in or on the reaction reservoir generates a flow pattern, the Mixing of the liquids leads, the flow pattern until the complete Homogenization of liquids is maintained. Preferably, a laminar flow pattern generated.

at the method according to the invention is the amount of the first liquid to be metered in the metering reservoir established. The first liquid will prevented from entering the reaction reservoir. In a preferred process management prevents the surface tension, that the liquid enters the reaction reservoir. Only when the first liquid with the second liquid comes in contact, which brought into or on the reaction reservoir was, can be a fluid exchange occur. In this case, due to the smaller cross section of the connecting channel structure the fluid exchange negligible due to diffusion. Only by generating a corresponding flow pattern in the reaction reservoir an effective mixing is effected. The amount of the second liquid is due to the size of the reaction reservoir certainly.

With the method according to the invention Can be a dosage and mixing of liquids in a large dynamic Range, so with very different mixing ratios precise carried out become. The mixing ratio between Reagents and sample fluid can be set between 1: 100 and 100: 1, for example.

The flow pattern can be due to sound waves in the liquid be generated on or in the second reservoir.

to Generation of sound waves can produce surface sound waves be used, in a conventional manner with the help of a Interdigitaltransducers are generated on a piezoelectric chip can, which is attached to the device. It will be either the Momentum transfer the surface acoustic waves directly or with the help of the surface acoustic wave generated sound waves used. The term surface sound waves includes in the present text also boundary sound waves at the interfaces between two solids.

The reservoirs and the connection structures can be designed in three dimensions or two dimensions. Thus, the reservoirs and interconnect structures may be correspondingly shaped depressions in a surface. Other configurations are correspondingly shaped cavities. In a two-dimensional embodiment, the reservoirs and connecting structures are formed by correspondingly shaped areas of a surface, which are wetted by the liquids more preferably than the surrounding areas of the surface. For aqueous solutions who chosen for the reservoirs and connecting structures surfaces that are hydrophilic compared to their environment. Such wetting-modulated surfaces are, for example, in DE 100 55 318 A1 described. The liquids are held together on the preferably wetted areas by their surface tension as drops.

to simpler representation in the present text are each three-dimensional and includes two-dimensional implementations, if not explicit otherwise stated, even if terms are chosen, the only one way seem to describe. This is also the case for applying, for example a liquid on a two-dimensional reservoir surface the term "introduction into a Reservoir "or" filling "is used for example also for the movement of liquid on a two-dimensional connection structure the term "movement through the Connection structure "used etc. The "volume" or size of a "cross-section" means in two-dimensional Realizations in an analogous way, for example, the area or the width.

at The connection structure may be an appropriately sized opening between the Dosagereservoir and the reaction reservoir act. A special precise Process management uses the capillary force in a connecting capillary structure, the from the first liquid is wetted and by the capillary forces from the Dosierreservoir filled becomes. At the entry point of the connecting capillary structure into the reaction reservoir take due to the enlarged cross-section the capillary forces abruptly, allowing leakage of the first fluid prevented from the Verbindungskapillarstruktur in the reaction reservoir becomes. Only when the second liquid is introduced into the reaction reservoir or applied to the reservoir surface becomes, the second liquid enters with the first liquid in conjunction so that mixing can occur.

at a three-dimensional metering device are in one embodiment the metering the reservoirs filled by filling, the preferably located in the upper end of the reservoir.

The Dosagereservoir can by an appropriately sized volume be formed. In a particularly preferred embodiment of the inventive method a reservoir capillary structure is used as the metering reservoir, which has at least two openings along its extent. Through an opening can filled the capillary structure become. liquid enters through the first opening and moves through the capillary force driven to the second opening. The Reservoirkapillarstruktur is doing so as a capillary structure selected, that the liquid front the moving liquid occupies the entire cross section of the capillary structure. Except the Filling opening and the second opening are no further openings in opened to the system. At the second opening stops the liquid her movement. Since no further vents are provided, it builds beyond the second opening a back pressure, which prevents further fluid movement. Furthermore decreases the capillary force at the second opening abruptly. A further filling over the second opening In addition, therefore, up to a certain threshold of the filling pressure not possible. This way is through the distance between the two openings defines an exact volume in the reservoir capillary structure, to allow an exact dosage. In a modification, two symmetrically arranged to the filling second openings used. The liquid volume the fluid dosed in such a reservoir capillary structure then corresponds to the distance between these two second openings.

A Continuing uses a Reservoirkapillarstruktur with several such selectable openings, depending on the desired Dosing volume of the first liquid open become. Will continue from the opening used as the filling opening removed openings open, so can the liquid up to these openings enter and a larger volume taking.

The Dosierreservoir corresponds to this process management the with first liquid filled Volume of the reservoir capillary structure. The remaining part of the Reservoir capillary structure is part of the reaction reservoir.

A Another advantageous embodiment of the method according to the invention employs a plurality of preferably different sized Dosierreservoirs via connecting structures communicate with the reaction reservoir. In addition, stand the Dosierreservoirs with a filling opening in connection. The connection structures between the individual metering reservoirs and the reaction reservoir can initially closed in one embodiment his and to choose the desired one Dosing reservoirs open become. In another embodiment, the desired metering reservoir with the desired Volume selected by that the rest Connecting structures closed to the other metering reservoirs become.

In a modification of this procedure, all dosing reservoirs are first filled and then the connection structure of the desired dosing reservoir is opened. In this case, if necessary, individual dosing reservoirs by other dosing filled reservoirs. Such a process management allows the filling of a larger number of Dosierreservoirs by only one filling opening and thus only a Stel development of a filling device, eg. B. a pipette tip. This procedure has the advantage that the corresponding filling device does not have to be moved and thus the apparatus required is low. Only after complete filling of all metering reservoirs, the selected metering reservoir is connected to the reaction reservoir by opening the corresponding connection structure.

Either the opening as well as the closing can with a suitable selection of the material of the dosing device used be effected by a melting process. So is suitable for example a plastic part as a metering device. Either the connection structures first sealed, with the desired connection structures before use are melted to make a connection. At a other proceedings dosing devices are used, in which the connection structures initially open are and are not needed Connecting structures before application by a melting process getting closed.

at another procedure especially for two-dimensional design will be the connection between the two liquids over one made small "bridgehead", the between the two liquids is brought and generates a liquid bridge. The bridge drop has doing a smaller volume than both the first and the second Amount of liquid.

Especially advantageous is an embodiment in which between a Dosierreservoir and the reaction reservoir is more than one connection structure. Here can be the fluid exchange - for example driven by sound waves - in take place a cycle until complete homogenization of the liquids occurred.

The inventive method is not on the metered addition of one fluid to a second amount of liquid limited. With a corresponding number of metering reservoirs and connection structures, which connect these metering reservoirs with the reaction reservoir, can several amounts of liquid at the same time or successively for addition to the liquid be provided in the reaction reservoir.

A device according to the invention, with the inventive method carried out can has at least one Dosierreservoir for a first amount of liquid on. Furthermore, a reaction reservoir for a second amount of liquid and at least one connection structure between the two reservoirs intended. The connection structure is preferably in proportion to the reservoirs dimensioned that the first liquid is not due to their surface tension can enter the reaction reservoir. Finally, the device according to the invention has a Device for generating preferably a laminar flow pattern for mixing of liquid in the reaction reservoir.

to Generation of the flow pattern shows a preferred embodiment at least one sound wave generating device for irradiation of Sound waves in the reaction reservoir on. Preferably, the at least one sound wave generating device by a surface acoustic wave generating device formed, in particular by an interdigital transducer on a piezoelectric chip.

The reservoirs and the at least one connecting structure may be formed as depressions or cavities in a solid body. In a two-dimensional embodiment of the device according to the invention, the reservoirs and connecting structures are formed by correspondingly shaped regions of a surface, which are more preferably wetted by the liquids than the surrounding regions of the surface. Such wetting-modulated surfaces are, for example, in DE 100 55 318 A1 described.

A three-dimensional embodiment the metering device according to the invention For example, it may include pits in a solid that are filled by a Lids are completed to the reservoirs or connecting structure to build. The lid can be easily made from a foil be, preferably made of plastic.

An apparatus according to the invention with which the method according to the invention can be carried out using a device according to the invention comprises a receptacle for a device according to the invention. When the device is inserted, the at least one device for generating a flow pattern is electrically contacted. The apparatus of the invention further comprises controllable filling devices, for. As pipettes or dispensers, which are arranged in the receptacle inserted device above the filling structures. By using a device according to the invention or carrying out the method according to the invention, the accuracy requirements for the filling devices are not very high, since the dosage only takes place within the device itself. Finally, the apparatus has a control for controlling the timing of a protocol, which controls the device to generate the flow pattern and the filling facilities. Preferred embodiments include opening means for opening individual filling structures, vents or barrier structures or means for closing individual barrier structures.

Of the inventive apparatus can with appropriate equipment also fulfill other functions, if z. B. a heater is provided for temperature control. After all can also z. B. electrical or optical evaluation with integrated.

With an apparatus according to the invention can the inventive method be carried out easily and automatically. It can as inventive devices disposable parts are easily used for integrated dosing and mixing become.

advantages the device according to the invention, the Apparatus according to the invention and preferred embodiments the dependent claims result from the above description of the advantages and preferred Embodiments of the method according to the invention.

The inventive method, the device according to the invention and the apparatus of the invention can especially effective for dosing and mixing biological liquids be used, where a precise dosing smallest amounts of liquid necessary is.

embodiments and embodiments of the invention will be apparent from the appended Figures explained in detail. The figures are not necessarily to scale and are used to schematic representation. It shows:

1 a top view of a metering device according to the invention in the open state,

2 a cross section in the direction of view II by the embodiment of the 1 .

3 a plan view of a further embodiment of a metering device according to the invention, and

4 a plan view of a third embodiment of the metering device according to the invention.

1 shows a plastic part 5 with chambers 1 . 3 , The plastic part 5 can be produced for example by injection molding. The cover of the chamber is covered by a thin laminated plastic film 2 causes the in 2 is visible and in 1 not shown to the inner workings of the plastic part 5 to clarify. The connection between the chambers 1 and 3 takes place via two bottlenecks 11 , reference numeral 13 refers to the wall between the chambers 1 and 3 ,

In 1 are the layers of the filling openings 7 and 9 indicated in the plastic film 2 are provided, however, in 1 not shown.

Below the chamber 1 , which is also referred to below as the reaction chamber, is an acoustic chip 15 , which may be, for example, a piezoelectric solid state chip, on which in known manner an interdigital transducer for generating surface acoustic waves is applied. The interdigital transducer is designed in such a way that the surface sound waves generated with it produce a sound wave radiation into the reaction chamber 1 enable. The emission of sound waves into a volume of liquid separated by a solid from the surface acoustic waves generating the interdigital transducer is shown in FIG DE 103 25 307 B3 described. In an analogous way, the acoustic chip 15 also on the slide 2 or be provided in a side area.

The acoustic chip 15 is connected via electrical connections, not shown, to an AC voltage source with which an AC voltage of a frequency of a few tens of MHz can be generated in order to produce with the interdigital transducer surface acoustic waves, which are used for the radiation of sound waves in the reaction chamber 1 to lead.

The location of the acoustic chip 15 is in 1 indicated, although the chip in this view would not be visible per se, since it is mounted in the embodiment shown on the underside of the device. In the sketch of the 1 the acoustic chip is drawn in the form of parallel lines, which only schematically the alignment of the individual finger electrodes of the interdigital transducer on the piezoelectric chip 15 to indicate. The emission direction of the surface acoustic waves of such an aligned interdigital transducer is perpendicular to the alignment of the finger electrodes.

The necessary size of the serving as a reaction reservoir chamber 1 depends on the frequency of the sound waves used. The smallest extent should be much larger than the wavelength of the sound used. Finally, the expansion of the reaction chamber should be 1 in the propagation direction of the sound waves about one Magnitude be greater than the extent of bottlenecks 11 , The smallest dimension of the reservoir is, for example, 1 mm to 10 mm with a sound wave length of, for example, 100 μm. The total length of the channel system is a few centimeters. The filling openings 7 . 9 are at least an order of magnitude smaller than the reaction chamber 1 ,

The device according to the invention of this embodiment is used as follows. The reaction reservoir comprises, for example, 100 .mu.l or 150 .mu.l while the metering reservoir holds 5 .mu.l. Such fluid volumes are particularly characteristic of many diagnostic applications. First, the dosing reservoir 3 through the filling hole 7 filled with a first liquid, which can be done for example by capillary force. The fluid gets to the bottlenecks 11 remain standing, because here the capillary force because of the larger diameter of the reservoir 1 abruptly decreases. Subsequently, the reservoir becomes 1 through the filling holes 9 filled with a second liquid. A possible supernatant of liquid on the respective filling holes 7 . 9 is not critical. The liquid of this supernatant does not participate in the following mixing process for geometrical reasons, in particular when the following mixing process is effected by a laminar flow pattern. In this way, the volumes of the two liquids have now been defined geometrically, without great precision of the filling devices used, for example, pipettes, would be necessary. At the bottlenecks 11 the liquids are in contact. Diffusion occurs due to the narrow cross-section of the bottlenecks 11 only to a negligible extent. A homogeneous mixing of the entire liquid quantities is done with the help of the acoustic chip 15 reached. By applying an AC voltage to the acoustic chip, acoustic energy is radiated into the defined volumes of the liquids and a laminar flow pattern is generated. The liquids or their ingredients are mixed and, if necessary, reacted. The result of this reaction can be read, for example, optically or electrically. It is advantageous that the filling holes 7 . 9 do not have to be closed.

The metering and mixing of the liquids thus takes place in a cost-effective optionally configured as a disposable cartridge device 5 , Dosing is also very easy. Even if it comes to a supernatant on the filling holes, this will not participate in the mixing reaction for geometric reasons and / or due to the laminar flow pattern used.

3 shows another embodiment of a metering device according to the invention. Shown here is the detail of a plastic body 105 containing the metering device, which also depressions in the plastic structure 105 includes. Visible is the reaction reservoir 101 with filling holes 109 , 103 shows a capillary structure with multiple openings, the opening 107 serves as a filling opening. The capillary structure 103 Fig. 10 illustrates a dosing capillary structure via connection capillary structures 111 with the reaction reservoir 101 communicates. The entire structure is also finished with a plastic film. Also in this embodiment are not visible in the open view openings 107 . 109 . 121 and 122 indicated to represent their relative position. Also indicated in its location is arranged below the device and therefore not actually visible in the representation acoustic chip 115 with an interdigital transducer. The acoustic chip 115 corresponds to the related to the 1 and 2 described chip 15 ,

With such a metering device, different mixing ratios can be set. The filling takes place via the filling hole 107 that is open. All other holes 109 . 121 . 122 are initially closed. The volume of the first liquid that is filled can now be opened by selectively opening the holes 121 . 122 to adjust. Are z. B. only one hole 121 in the immediate vicinity of the filling opening 107 and the hole arranged symmetrically on the other side 121 opened, it is possible to define a liquid volume of a length equal to the distance between the two opened openings 121 equivalent.

The capillary structure 103 causes the front of the liquid the entire cross section of the capillary structure 103 fills. If no other vent holes are opened, builds up a back pressure, which leads to the stopping of the liquid. A movement over the open holes 121 It is therefore not possible. This effect is reinforced by the fact that through the opened opening 121 the capillary force causing the movement decreases.

Be the two outer openings 122 opened, results in a correspondingly larger volume.

In both cases, the residual volume in channel 103 and the connection capillary structures 111 by filling via the reaction reservoir 101 through the openings then open 109 be filled. The residual volume of the channel 103 then counts to the reaction reservoir.

The characteristic dimensions of an embodiment according to the 3 correspond to the characteristic dimensions of the embodiment of the 1 and 2 ,

With such an embodiment, therefore, the setting of different mixing ratios in a simple manner possible. Depending on how much of the first liquid to be added to the second liquid, the corresponding openings 121 . 122 open. This can be done, for example, by simply piercing the plastic film at accordingly marked locations. The further mode of operation essentially corresponds to the embodiment of FIG 1 and 2 ,

4 shows another embodiment. Here are several dosing reservoirs 203 . 223 provided by connecting capillary structures 211 . 212 with the reaction reservoir 201 keep in touch. The dosing reservoirs 203 . 223 have different sized volumes and have a connection channel structure 216. in connection. In the connection channel structure 216. is the filling opening 207 , The dosing reservoirs 203 . 223 have vents 221 on. The connection channel 216. in the embodiment shown is via a connecting capillary structure 210 also with the reaction reservoir 201 connected. The structure 210 also includes a vent hole 221 , Finally, in the reaction reservoir 201 filling openings 209 intended.

217 . 218 . 219 . 220 and 224 schematically represent barrier structures. The entire metering device of 4 is provided in a plastic part that passes through a foil with openings 207 . 209 . 221 is completed. It may be in the metering of the 4 also a disposable part, which is prefabricated ex works. In this case, in the embodiment shown, the barrier structures 217 . 218 . 219 . 220 . 224 initially closed.

Also in the presentation of 4 are the filling openings not visible in the open representation 207 . 209 or the vents 221 indicated in their position. In addition, under the arrangement of the 4 an acoustic chip 215 , the acoustic chip already described 15 . 115 equivalent. Also the acoustic chip 215 is in 4 indicated, although it is not visible in this representation, since it is located below the arrangement.

Also in the embodiment of the 4 The characteristic dimensions correspond to the characteristic dimensions of the embodiment of FIG 1 and 2 ,

For use of the embodiment, it is first decided in a process management which of the metering reservoirs 203 . 223 to be filled with liquid to define a corresponding volume of liquid. For explanation, in the present description, the metering reservoir 223 selected. After the selection is made, the corresponding barriers will be 217 . 219 connected to the dosing reservoir 223 bordered, for example, by a heater or with laser energy melted. This can be done, for example, with the aid of a machine that processes the dosing device.

The appropriately selected dosing reservoir 223 can then over the filling opening 207 be filled and used for dosing. In this case, the dosage is similar to, for example, in the embodiment of 1 and 2 described carried out. In particular, the dimensions of the structures are selected so that filling of the metering reservoir can be effected by the action of the capillary force. Alternatively, a filling can be done with pressure. The vent 221 is arranged so that a complete filling of the reservoir is possible.

Due to the entry point of the connection capillary structure 211 in the reaction reservoir 201 abruptly decreasing capillary action, the liquid does not enter the reaction reservoir 201 one. Only when filling the reaction reservoir 201 through the filling openings 209 Liquid comes out of the reaction reservoir 201 with liquid in the connec tion capillary structure 211 in connection. The further operation in turn corresponds to the embodiment of the 1 and 2 ,

Will the reservoir 203 is selected analogously using the corresponding barrier structures 218 . 220 and the connection capillary structure 212 method.

Another embodiment of this embodiment does not include any barrier structures ex works 217 . 219 , Before the application, it is first decided again which of the metering reservoirs 203 . 223 should be used. If z. B. the metering reservoir 223 selected, becomes the other dosing reservoir 203 decoupled with the help of a machine, in the places of the barriers 218 . 220 , which is the unused dosing reservoir 203 are adjacent, by application of heating energy or laser energy the appropriate connection channel structures zuschmilzt.

Also in the embodiments according to the 4 can the individual Dosierreservoirs 203 . 223 in each case via a plurality of connecting capillary structures 211 . 212 with the reaction reservoir 201 be connected, which are open when selecting the appropriate Dosierreservoirs.

In addition to the connection structures 211 . 212 with the barrier structures 219 . 220 may be another Verbindungskapapillarstruktur 210 be provided, which the connecting channel 216. with the reaction reservoir 201 combines. Also this connection capillary structure 210 includes a vent 221 and optionally a barrier structure 224 , The additional channel 210 can be used to form a circuit that promotes effective mixing. After one of the dosing reservoirs 203 . 223 is selected, it is filled. For the purpose of the description, this again is the dosing reservoir 223 , First, an embodiment will be described in which the barrier structures 217 . 218 . 219 . 220 . 224 are initially closed. To fill the reservoir 223 becomes the barrier structure 217 melted as described. Through the filling opening 207 Liquid is introduced, which is the dosing reservoir 223 and the connection capillary structure 211 crowded. Also the connection capillary structure 210 is filled with this liquid. The filling takes place for example by capillary force.

Now the barrier structures can 219 . 224 be melted. The liquid enters at the entry points of the connection capillary structures due to the 211 . 210 in the reservoir 201 abruptly decreasing capillary action not in the reservoir 201 one. Filling the reservoir 201 through the openings 209 with a second liquid causes the contact of the liquids at the entry points of the connecting capillary structures 210 . 211 , Creating a laminar flow, for example, with the acoustic chip 215 then causes an effective mixing of the liquids. This can lead to a circulatory movement of the liquids.

In such an embodiment, utilizing capillary forces in the connecting capillary structures 210 . 211 . 212 can on the barrier structure 224 be completely dispensed with. Especially in one embodiment with only two Dosierreservoirs, as in 4 is shown, takes the Verbindungskapapillarstruktur 210 In any case, participate in the cycle process, so that decoupling is not necessary.

In another procedure, the barrier structures become 219 . 224 only after introduction of the second liquid into the reservoir 201 melted. Otherwise the procedure is the same. In such a process management, the connection structures 210 . 211 . 212 not necessarily be able to exert capillary action on the liquids.

Another process using an apparatus according to 4 uses barrier structures 217 . 218 . 219 . 220 . 224 that are originally open. Through the filling opening 207 first liquid is introduced. This flows due to the capillary action in the metering reservoirs 203 . 223 and in the connection capillary structures 210 . 211 . 212 , They do not enter the reaction reservoir 201 a, because at the entry points of the connection structures 210 . 211 . 212 in the reaction reservoir 201 the capillary action breaks off. It is only now decided which dosing reservoir, and therefore which dosing volume of the first liquid, should be used. For the purpose of the present description, this again is the metering reservoir 223 , The barrier structures 218 . 220 are then sealed as described and thus the unused Dosierreservoir 203 decoupled with the liquid contained therein. It then becomes second liquid in the reaction reservoir 201 fills. The subsequent procedure corresponds to the already described cycle procedure.

A inventive device can also have more than two dosing reservoirs with corresponding connection structures include. It can then several Dosierreservoirs in the circuit "in series" are switched to the dosing the first liquid to enlarge. at Such a configuration can the individual dosing reservoirs have different or the same size.

specially in a procedure, in the case of a cycle of fluids is used, finds a reaction between the liquids not only in the reaction reservoir part of the device instead of.

to Differentiation against the use of the term "dosing reservoir", with which the dosage of the first liquid the term "reaction reservoir" has nevertheless been used in the present text since, in particular, in the embodiments shown the reaction reservoir is the main structure due to its size, in the reaction takes place. In particular, for example, at a embodiment of the

1 or the 4 However, it is also possible that the Dosierreservoirs and the reaction reservoir z. B. are the same size and in a circulation process also takes place in both reservoirs a reaction.

The dosing or mixing devices according to the invention can be processed in an automatic machine which fills the liquids into the devices, tempering the devices, controlling the acoustic chips and also opening or opening or closing the filling holes. In addition, if necessary, with such a machine, the z. B. electrical or optical evaluation are made. Such machines can be useful in diagnostics or general laboratory automation.

Therefore, it may be advantageous, for example, if in the embodiments of the 3 and 4 the filling of the reservoirs takes place only through one or at most two filling holes, since this simplifies the addition of the liquid through the machine. Corresponding pipetting heads or dispensers for filling can then be made stationary.

With the embodiments shown can for example, total volumes of up to 1 ml for single volumes of z. B. only 100nl be processed.

With the method according to the invention Can be a dosage and mixing of liquids in a large dynamic Range, so with very different mixing ratios precise carried out become. The requirements for the accuracy of the filling devices used is not high, since the dosage by the process of the invention or Use of the device according to the invention happens. The mixing ratio between reagents and sample fluid can be set between 1: 100 and 100: 1, for example.

1

Reaction reservoir, reaction chamber

2

Plastic film

3

metering reservoir, metering

5

Plastic part

7, 9

filling openings

10

contraption for integrated dosing and through

Mix

11

bottlenecks

13

boundary wall

15

acoustic chip

20 30

devices for integrated dosing and

by mixing

101

Reaction reservoir, reaction chamber

103

Dosierkapillarstruktur

105

Plastic part

107 109

filling openings

111

Verbindungskapillarstruktur

115

acoustic chip

121 122

openings

201

Reaction reservoir, reaction chamber

203

metering reservoir, metering

207 209

filling openings

210 211, 212

Verbindungskapillarstrukturen

215

acoustic chip

216

Connecting channel structure

217 218, 219, 220

barrier structures

221 222

vents

223

metering reservoir, metering

224

barrier structure

Claims (41)

Method for integrated dosing and mixing of small quantities of liquid with a volume of up to 1 ml, in which at least one dosing reservoir ( 3 . 103 . 203 . 223 ) is completely filled with a first liquid, which via at least one connecting structure ( 11 . 111 . 211 . 212 ) with a reaction reservoir ( 1 . 101 . 201 ), wherein the connecting structure is designed such that the entry of the first liquid into the reaction reservoir ( 1 . 101 . 201 ) is first prevented, a reaction reservoir ( 1 . 101 . 201 ) is completely filled with a second liquid, so that the second liquid at the connection structure ( 11 . 111 . 211 . 212 ) comes into contact with the first liquid, and in the liquid at least in or on the reaction reservoir ( 1 . 101 . 201 ), a flow pattern is generated, which leads to the mixing of the liquids. Method according to Claim 1, in which the connection structure ( 11 . 111 . 211 . 212 ) is dimensioned such that the surface tension of the first liquid entering the reaction reservoir ( 1 . 101 . 201 ) prevented. Method according to one of claims 1 or 2, wherein for mixing a substantially laminar flow pattern is generated. Method according to one of claims 1 to 3, wherein for mixing sound waves in the liquid in or on the reaction reservoir ( 1 . 101 . 201 ) are irradiated. Method according to claim 4, wherein the sound waves with surface sound waves preferably with the aid of at least one interdigital transducer be generated. Method according to one of claims 1 to 5, wherein the at least a connection structure comprises a connection capillary structure. Method according to one of Claims 1 to 6, in which the reservoirs and the at least one connecting structure are formed by areas on a surface which, in comparison with the surrounding surface of the liquids are preferably wetted. Method according to one of claims 1 to 6, wherein the reservoirs and the at least one connection structure through recesses in a surface be formed. Method according to one of Claims 1 to 6, in which the reservoirs ( 1 . 3 . 101 . 103 . 201 . 203 . 223 ) and the at least one connection structure ( 11 . 111 . 210 . 211 . 212 ) through cavities in a solid state structure ( 5 . 105 ) are formed. Method according to Claim 9, in which the reservoirs ( 1 . 3 . 101 . 103 . 201 . 203 . 223 ) through filling openings ( 7 . 9 . 107 . 109 . 207 . 209 ) are preferably filled in the upper end of the reservoirs. Method according to one of Claims 9 or 10, in which a reservoir capillary structure ( 103 ) is used, which along its longitudinal extent at least two openings ( 107 . 121 . 122 ) having. Method according to Claim 11, in which the volume of the first quantity of liquid is determined by the selection of the openings ( 121 . 122 ) in the reservoir capillary structure ( 103 ) is selected. Method according to one of Claims 1 to 12, in which a device with a plurality of metering reservoirs ( 203 . 223 ) is preferably used of different sizes, on the one hand via connection structures ( 211 . 212 ) with the reaction reservoir ( 201 ) and on the other hand with a filling opening ( 207 ) keep in touch. Method according to Claim 13, insofar as it depends directly or indirectly on one of Claims 8 or 9, in which the connection structures ( 211 . 212 ) are initially closed and to select the desired Dosierreservoirs ( 203 . 223 ). Method according to claim 13, insofar as it depends directly or indirectly on one of claims 8 or 9, in which the desired dosing reservoir ( 203 . 223 ) by closing the connection structures ( 211 . 212 ) is selected to the other Dosierreservoirs. Method according to one of claims 14 or 15, in which the opening or closing of the connecting structures ( 211 ) is effected by a melting process in a plastic part. Device for the integrated dosing and mixing of small quantities of liquid with a volume of up to 1 ml for carrying out the method according to claim 1, with at least one dosing reservoir ( 3 . 103 . 203 . 223 ) for a first quantity of liquid, a reaction reservoir ( 1 . 101 . 201 ) for a second quantity of liquid, at least one connecting structure ( 11 . 111 . 211 . 212 ) between the at least one metering reservoir and the reaction reservoir, wherein the connecting structure is configured such that the first liquid does not enter the reaction reservoir ( 1 . 101 . 201 ), and a body ( 15 . 115 . 215 ) for generating a flow pattern for mixing liquid at least in or on the reaction reservoir. Device according to Claim 17, in which the connection structure ( 11 . 111 . 211 . 212 ) is dimensioned such that the first liquid due to their surface tension not in the reaction reservoir ( 1 . 101 . 201 ) entry. Device according to one of claims 17 or 18, in which the device for generating a flow pattern comprises at least one sound wave generating device ( 15 . 115 . 215 ) for the irradiation of sound waves in the reaction reservoir ( 1 . 101 . 201 ). Apparatus according to claim 19, wherein said at least one sound wave generating means comprises surface acoustic wave generating means (16). 15 . 115 . 215 ). Apparatus according to claim 20, comprising at least one Interdigital transducer for generating surface acoustic waves. Device according to one of claims 17 to 21, in which the at least one connection structure has a connection capillary structure includes. Device according to one of claims 17 to 22, wherein the Reservoirs and the at least one connection structure surface areas on a surface include, compared to the surrounding surface of the fluids preferably be wetted. Device according to one of claims 17 to 22, wherein the Reservoirs and the at least one connecting structure depressions in a surface. Device according to one of Claims 17 to 22, in which the reservoirs ( 1 . 3 . 101 . 103 . 201 . 203 . 223 ) and the at least one connection structure ( 11 . 111 . 210 . 211 . 212 ) Cavities in a solid state structure ( 5 . 105 ). Apparatus according to claim 25, wherein the cavities are recesses in a solid ( 5 . 105 ) provided by a lid ( 2 ) Are completed. Device according to Claim 26, in which the lid ( 2 ) through a foil ( 2 ), preferably made of plastic. Device according to one of claims 25 to 27, wherein the at least one metering reservoir comprises a metering capillary structure ( 103 ). Device according to one of claims 25 to 28, wherein the cavities predetermined openings ( 121 . 122 ), which are initially closed and can be opened if necessary. Device according to claims 28 and 29, wherein the metering capillary structure ( 103 ) at least two predetermined, possibly openable openings ( 121 . 122 ) arranged along the dosing capillary structure. Device according to one of claims 17 to 27, wherein a plurality of metering reservoirs ( 203 . 223 ), preferably of different sizes, which are provided via connecting structures ( 211 . 212 ) with the reaction reservoir ( 201 ) are connected, which are initially closed and can be opened if necessary. Device according to claim 31, insofar as it depends directly or indirectly on one of the claims 24 or 25, in which the connection structures ( 211 . 212 ) fusible barriers ( 219 . 220 ) comprise plastic. Device according to one of claims 17 to 27, wherein a plurality of metering reservoirs ( 203 . 223 ), preferably of different sizes, which are provided via connecting structures ( 211 . 212 ) with the reaction reservoir ( 201 ) are connected, which are initially open and can be closed if necessary. Apparatus according to claim 33, insofar as it depends directly or indirectly on one of claims 24 or 25, in which the connection structures ( 211 . 212 ) Include barrier structures that become barriers through a melt process ( 219 . 220 ) can be formed. Apparatus for automatically carrying out a method according to claim 1, with a receptacle for a device ( 10 . 20 . 30 ) according to claim 17, electrical contacts for contacting the at least one device ( 15 . 115 . 215 ) for generating a flow pattern which, when inserted in the receptacle device ( 10 . 20 . 30 ) the at least one device ( 15 . 115 . 215 ) electrically contact to produce a flow pattern, means for automatically supplying liquid into the reservoirs ( 1 . 3 . 101 . 103 . 201 . 203 . 223 ), and a controller, preferably comprising a microprocessor, for controlling the at least one device ( 15 . 115 . 215 ) for generating a flow pattern and the automatic liquid supply means. Apparatus according to Claim 35, in which the automatic liquid supply devices comprise electrically activatable pipetting tips and / or dispensers which, when the device is inserted in the receptacle ( 10 . 20 . 30 ) above corresponding filling openings ( 7 . 9 . 107 . 109 . 207 . 209 ) are arranged. Apparatus according to one of claims 35 or 36, with opening-controllable opening means ( 121 . 122 ) of a device inserted in the receptacle ( 20 ) according to one of claims 29 or 30. Apparatus according to claim 37, in which the opening means are piercing points for piercing one of the lids of the device ( 20 ) include plastic film. Apparatus according to one of claims 35 or 36, comprising heat-generating means controllable by the controller for generating heat at the locations of the barrier structures ( 217 . 218 . 219 . 220 . 224 ) a device ( 30 ) according to one of claims 32 or 34. Apparatus according to claim 39, wherein the heat generating means heating wires or laser. Use of a method according to one of claims 1 to 16, a device ( 10 . 20 . 30 ) according to any one of claims 17 to 34 or an apparatus according to any one of claims 35 to 40 for the metering and mixing of biological fluids.