Microfluidic Device for Processing and Aliquoting a Sample Liquid, Method and Controller for Operating a Microfluidic Device, and Microfluidic System for Carrying Out an Analysis of a Sample Liquid

Abstract

A microfluidic device is for processing and aliquoting a sample liquid. The microfluidic device has a dividing chamber for receiving a starting volume of the sample liquid. The dividing chamber has a plurality of cavities for receiving sub-volumes of the sample liquid, the sub-volumes being usable for analytical reactions. The microfluidic device also has a microfluidic network for using the dividing chamber in a fluid-mechanical manner and at least one pump device for pumping fluids within the device. The at least one pump device and the microfluidic network are configured to pump the sample liquid, as a first phase, and a sealing liquid, as a second phase, through the microfluidic network and into the dividing chamber in order to seal the sub-volumes of the sample liquid in the cavities using the sealing liquid.

Claims

1. A microfluidic apparatus for processing and aliquoting a sample liquid, the microfluidic apparatus comprising: a division chamber configured to accommodate an input volume of the sample liquid, the division chamber defining a plurality of cavities configured to accommodate partial volumes of the sample liquid that are usable for detection reactions; a microfluidic network configured to make the division chamber accessible in fluid-mechanical fashion, the microfluidic network defining at least one feed channel and a removal channel connected to the division chamber in fluid-mechanical fashion; and at least one pump device configured to convey fluids within the microfluidic apparatus, wherein the at least one pump device and the microfluidic network are configured to convey the sample liquid as a first phase through the microfluidic network into the division chamber, in order to arrange the partial volumes of the sample liquid in the cavities of the plurality of cavities, and to convey a sealing liquid as a second phase through the microfluidic network into the division chamber, in order to seal the partial volumes of the sample liquid in the cavities of the plurality of cavities using the sealing liquid.

2. The microfluidic apparatus as claimed in claim 1, further comprising: at least one channel branching point of the at least one feed channel configured to branch into a discharge channel and a supply channel, the supply channel connected to the division chamber in fluid-mechanical fashion; and at least one valve configured to influence a fluid flow in a region of the channel branching point.

3. The microfluidic apparatus as claimed in claim 1, further comprising: the sample liquid; and the sealing liquid.

4. The microfluidic apparatus as claimed in claim 1, further comprising: a temperature-control device configured to control a temperature of the partial volumes of the sample liquid that are arranged in the cavities; and/or a detection device configured to optically detect at least one property of the partial volumes of the sample liquid that are arranged in the cavities.

5. The microfluidic apparatus as claimed in claim 2, wherein: the supply channel is branched into at least two sub-channels which lead into the division chamber, and at least one dimension of a fluid channel cross section is reduced at a region in which the at least two sub-channels lead into the division chamber.

6. The microfluidic apparatus as claimed in claim 1, wherein: the cavities of the plurality of cavities are formed in a chip which is arranged in the division chamber, and at least one dimension of a fluid-conducting region of the division chamber is reduced in a transition region to the chip in the division chamber.

7. The microfluidic apparatus as claimed in claim 2, further comprising: at least one elastic membrane configured to be (i) deflected into at least one pump chamber in order to perform a function of the at least one pump device, and/or (ii) deflected into at least one valve chamber in order to perform a function of the at least one valve.

8. The microfluidic apparatus as claimed in claim 1, wherein: the at least one pump device includes a plurality of the pump devices, and the pump devices configured to convey the fluid in the microfluidic network at different flow rates and/or to convey different fluid volumes per pump cycle.

9. The microfluidic apparatus as claimed in claim 1, further comprising: a further chamber connected in parallel to the at least one feed channel in fluid-mechanical fashion and connected to a ventilation channel in fluid-mechanical fashion; and a further temperature-control device configured to control a temperature of fluid arranged in the further chamber.

10. A method for operating a microfluidic apparatus comprising: introducing a sample liquid into the microfluidic apparatus; effecting conveyance of the sample liquid as first phase through a microfluidic network into a division chamber in order to arrange partial volumes of the sample liquid in cavities of a plurality of cavities; and effecting conveyance of a sealing liquid as a second phase through the microfluidic network into the division chamber in order to seal the partial volumes of the sample liquid in the cavities using the sealing liquid.

11. The method as claimed in claim 10, wherein effecting h conveyance of the sample liquid and effecting the conveyance of the sealing liquid comprises: producing a multi-phase system from the sample liquid as first phase and from at least one further phase, which comprises the sealing liquid and a transport liquid, in the microfluidic network; transporting the multi-phase system via a feed channel to a channel branching point using the at least one pump device, wherein at least one valve is controlled such that h transport liquid discharged via a discharge channel; and introducing the sample liquid, followed by the sealing liquid, via a supply channel into the division chamber by switching over the at least one valve after a boundary interface between the sample liquid and the transport liquid has passed the channel branching point.

12. The method as claimed in claim 10, further comprising: controlling a the temperature of the partial volumes of the sample liquid that are arranged in the cavities.

13. The method as claimed in claim 10, further comprising: optically detecting at least one property of the partial volumes of the sample liquid that are arranged in the cavities.

14. The method as claimed in claim 10, further comprising: thermally degassing the sample liquid and/or the sealing liquid in a further chamber which is connected in parallel to the at least one feed channel in fluid-mechanical fashion and is connected to a ventilation channel in fluid-mechanical fashion.

15. The method as claimed in claim 14, further comprising: displacing the sealing liquid which seals the partial volumes of the sample liquid that are arranged in the cavities by the sealing liquid that has been thermally degassed.

Description

[0048] Exemplary embodiments of the approach presented here are illustrated in the drawings and discussed in more detail in the following description. In the drawings:

[0049] FIG. 1 shows a schematic illustration of a microfluidic apparatus according to one exemplary embodiment;

[0050] FIG. 2A shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0051] FIG. 2B shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0052] FIG. 2C shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0053] FIG. 3 shows a schematic illustration of a microfluidic apparatus according to one exemplary embodiment;

[0054] FIG. 4 shows a schematic illustration of a microfluidic apparatus according to one exemplary embodiment;

[0055] FIG. 5A shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0056] FIG. 5B shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0057] FIG. 5C shows a schematic illustration of a partial portion of a microfluidic apparatus according to one exemplary embodiment;

[0058] FIG. 6 shows a schematic illustration of a microfluidic apparatus according to one exemplary embodiment; and

[0059] FIG. 7 shows a flow diagram of an operating method according to one exemplary embodiment.

[0060] In the following description of expedient exemplary embodiments of the present invention, the same or similar reference designations will be used for the elements of similar action illustrated in the various figures, wherein a repeated description of these elements will be omitted.

[0061] FIG. 1 shows a schematic illustration of a microfluidic apparatus 100 according to one exemplary embodiment, in particular a schematic illustration of a cross section through a microfluidic apparatus 100 according to one exemplary embodiment. A microfluidic network is connected to a central chamber or division chamber 115 via at least one feed channel 111, at least one pump device 121, and at least one channel branching point 114 of the feed channel 111 into a discharge channel 112 and a supply channel 113, and at least two valves 131, 132 or alternatively a multi-way valve for controlling the microfluidic flow at the branching point 114.

[0062] The division chamber 115 has in particular a plurality of cavities or apertures or compartments 140 which can be filled with a sample liquid 10 as first phase and can be overlaid with a sealing liquid 20 as second phase, such that the sample liquid 10 at least partially remains in the cavities 140. In this way, microfluidic aliquoting of the sample liquid 10 is achieved. Furthermore, the division chamber 115 also has a connection to a removal channel 116 in addition to a connection to the supply channel 113.

[0063] In other words, the microfluidic apparatus 100 for processing and aliquoting the sample liquid 10 thus comprises the division chamber 115 for the purpose of accommodating an input volume of the sample liquid 10. The division chamber 115 has a plurality of cavities 140 for accommodating partial volumes of the sample liquid 10 that are usable for detection reactions. Furthermore, the apparatus 100 comprises a microfluidic network for making the division chamber 115 accessible in fluid-mechanical fashion. The microfluidic network has at least one feed channel 111 having at least one channel branching point 114 into a discharge channel 112 and a supply channel 113 which is connected to the division chamber 115 in fluid-mechanical fashion, at least one valve 131, 132 for influencing a fluid flow in the region of the channel branching point 114 and a removal channel 116 which is connected to the division chamber 115 in fluid-mechanical fashion. Furthermore, the apparatus 100 comprises at least one pump device 121 for conveying fluids within the apparatus 100. The at least one pump device 121 and the microfluidic network are designed to convey the sample liquid 10 as a first phase through the microfluidic network into the division chamber 115, in order to arrange partial volumes of the sample liquid 10 in the cavities 140, and to convey a sealing liquid 20 as a second phase through the microfluidic network into the division chamber 115, in order to seal the partial volumes of the sample liquid 10 in the cavities 140 using the sealing liquid 20.

[0064] In the exemplary embodiment illustrated schematically in FIG. 1, the apparatus 100 additionally comprises at least one thermal interface or heat-exchange interface or temperature-control device 201 in the region of the division chamber 115 and in particular of the cavities 140, and also an optical interface or detection device 301 in particular in the region of the cavities 140. The temperature-control device 201 can thus be used in particular to control the temperature of the first phase or sample liquid 10 enclosed in the cavities 140. The detection device 301 can be used in particular to optically read a fluorescence signal which is emitted in particular by the sample liquid 10 enclosed in the cavities 140. Furthermore, during the processing, the apparatus 100 in the exemplary embodiment shown in FIG. 1 is suitably oriented with respect to a gravitational field g or alternatively set in rotation, such that a buoyancy force 500 results which can be used to discharge gas bubbles 50 that may form.

[0065] According to the exemplary embodiment illustrated in FIG. 1, the pump device 121 is connected in the feed channel 111 in fluid-mechanical fashion. A first valve 131 is connected in the supply channel 113 between the branching point 114 and the division chamber 115. A second valve 132 is connected in the discharge channel 112.

[0066] FIG. 2A, FIG. 2B and FIG. 2C show schematic illustrations of a partial portion of an apparatus according to one exemplary embodiment. The apparatus corresponds or is similar to the apparatus from FIG. 1. FIG. 2A shows an oblique plan view, FIG. 2B shows a plan view and FIG. 2C shows a sectional view of the partial portion of the apparatus. In this exemplary embodiment, the cavities 140 are located in a chip which is fixed in the division chamber 115, for example by means of an adhesive bond which connects a first side of the chip and a first side of the division chamber 115 to one another.

[0067] The supply channel 113 leads from the first side into the division chamber 115. The removal channel 116 is arranged on a second side of the division chamber 115. The geometry of the division chamber 115 and of the chip comprising the cavities 140 results in an abrupt reduction in the spatial dimensions 1130, 1150 of the fluid-conducting region of the division chamber 115 at the transition to the chip comprising the cavities 140. This reduction in the spatial dimensions 1130, 1150 is accompanied by a change in the capillary pressure that is present in accordance with the Young-Laplace equation. So-called pinning also occurs at an edge which is present at the location of abrupt reduction in the fluid-conducting region. In this way, an alignment, assisted by capillary action, of a liquid meniscus along the total width of the chip can be promoted, before the liquid wets a second side of the chip comprising the cavities 140. The spatially homogeneous change in capillary pressure and fluidic resistance along the total width of the chip also assists in the formation of a homogeneous flow profile in the division chamber 115, in particular in the region of the cavities 140 which are arranged on the second side of the chip.

[0068] In addition, in this advantageous configuration of the apparatus, the use of a sealing liquid having a density higher than the density of the sample liquid, the introduction of the liquids on the first side of the central chamber 115 and a suitable alignment of the central chamber 115 and/or of the apparatus 100 with respect to a gravitational field, for example by suitable tilting of the apparatus, make it possible, on account of the present density difference, to achieve a stable separation of sample liquid and sealing liquid and a spatially uniform propagation of the two-phase boundary interface through the central chamber 115, in the case of which each of the cavities 140 is first filled with sample liquid and then overlaid with the sealing liquid.

[0069] Overall, in dependence on the selected dimensions, the apparatus thus permits the formation of a flow profile that is as spatially homogeneous as possible both as a result of the arising capillary forces and the gravitational force acting on the liquids. In this way, on the one hand, reliable filling and sealing of all the cavities 140 can be achieved and, on the other hand, a high transfer efficiency of the sample liquid from the microfluidic network into the cavities 140 of the aliquoting structure can be obtained; i.e. a relatively small volume of sample liquid is already sufficient for filling all the cavities 140.

[0070] FIG. 3 shows a schematic illustration of a microfluidic apparatus 100 according to one exemplary embodiment, in particular a schematic cross section through an apparatus 100 according to a further exemplary embodiment. Here, the apparatus 100 is similar to the apparatus from one of the figures outlined above, in particular FIG. 1. In this exemplary embodiment, the apparatus 100 comprises two pump devices 121, 122, such as for example peristaltic pumps, which are suitable for effecting different flow rates in the microfluidic network of the apparatus 100. The combination of two pump devices 121, 122 having different pump volumes makes it possible to achieve both particularly rapid and particularly precise pumping of liquids. Furthermore, in the exemplary embodiment illustrated in FIG. 3, the supply channel 131 to the central chamber 115 has a branching arrangement 1131 which is used for the production of a spatially homogeneous flow in the central chamber 115 and for the capillary stabilization of the microfluidic boundary interfaces during the widening of the flow.

[0071] Here, a second pump device 122 is connected in the feed channel 111 between a first pump device 121 and the branching point 114. At the branching arrangement 1131, the supply channel 113 branches into a plurality of sub-channels, here four sub-channels merely by way of example.

[0072] FIG. 4 shows a schematic illustration of an apparatus 100 according to one exemplary embodiment. Here, the apparatus 100 is similar to the apparatus from one of the figures outlined above. In this exemplary embodiment of the apparatus 100, production and control of a microfluidic flow is based on the use of an elastic membrane which can be deflected by targeted application of pressure at defined points. The membrane is deflected into apertures of the microfluidic network that are provided therefor in order to, as a result, for example displace liquids, e.g. in the form of a pump chamber, or to open or close a fluidic path, e.g. in the form of at least one valve. In the exemplary embodiment of the apparatus 100 illustrated in FIG. 4, three microfluidic valves are arranged on the supply channel 111, which form a peristaltic pump unit 121. The combination of two of the aforementioned three valves of the supply channel 111 with the pump chamber adjoining the two valves has the effect of realizing a second pump function 122. In dependence on the pump function used, it is possible to transfer different volumes in a pump cycle. On the left below the central chamber 115 in the perspective projection depicted in FIG. 4, the supply channel 111 has a branching point 114 into a connecting channel 113 to the central chamber 115 and a discharge channel 112. The connecting channel 113 has a two-stage branching arrangement 1131 prior to the introduction into the central chamber 115 comprising the cavities 140. The central chamber 115 also has a removal channel 116.

[0073] FIG. 5A, FIG. 5B and FIG. 5C show schematic illustrations of a partial portion of a microfluidic apparatus according to one exemplary embodiment. Here, the apparatus corresponds or is similar to the apparatus from FIG. 4. FIG. 5A shows an oblique plan view, FIG. 5B shows a plan view and FIG. 5C shows a sectional view of the partial portion of the apparatus.

[0074] More precisely, this is an implementation of the division chamber 115 comprising an aliquoting structure composed of cavities 140, said division chamber being connected to a microfluidic network via a supply channel 113 having a branching arrangement 1131 and a removal channel 116. In this advantageous embodiment of the apparatus according to the invention, there is a reduction in the spatial dimensions 1130, 1150 of the fluid-conducting structures at the transition of the, here for example, four channels 1132 of the branching arrangement 1131 to the division chamber 115. In particular, a height 1150 of the division chamber 115 is significantly smaller than an extent 1130 of the supply channels 1132 of the branching arrangement 1131 at the transition to the division chamber 115. In accordance with the Young-Laplace equation, this corresponds with a change in the capillary pressure that is present at the transition of the supply channels 1132 to the division chamber 115, such that the “pinning” of phase boundary interfaces that occurs here has the effect that the channels 1132 of the branching arrangement 1131 can first be completely filled and then the division chamber 115 can be filled as homogeneously as possible.

[0075] FIG. 6 shows a schematic illustration of a microfluidic apparatus 100 according to one exemplary embodiment, in particular a schematic cross section through an apparatus 100 according to a further exemplary embodiment. Here, the apparatus 100 is similar to the apparatus from FIG. 3. Differences between the apparatus from FIG. 3 and the apparatus 100 illustrated in FIG. 6 are discussed below.

[0076] According to the exemplary embodiment illustrated here, the apparatus 100 comprises a further chamber 117 which is connected to the microfluidic network and which has a ventilation channel 118. Furthermore, the apparatus 100 comprises a further temperature-control device or thermal interface or heat-exchange interface 202 in the region of the further chamber 117. As a result, the further chamber 117 can be used in particular to control the temperature of liquids 10, 20, 30, for example for thermal degassing. The ventilation channel 118 makes it possible in particular to discharge gas bubbles 50 that form. The microfluidic channels 110, 111, 112, 113, 116, the pump devices 121, 122, 123 and the valves 130, 131, 132 can in this case be used to produce and control the microfluidic flow in a suitable manner, in particular between the division chamber 115, the further chamber 117 and the microfluidic network within the apparatus 100.

[0077] The first pump device 121 is connected in the feed channel 111 in fluid-mechanical fashion between the second pump device 122 and a third pump device 123. Here, the second pump device 122 is arranged between the first pump device 121 and the branching point 114. The ventilation channel 118 can be ventilated or shut off by means of a valve 130. The further chamber 117 is connected via a further channel 110 to the feed channel 111 between the second pump device 122 and the branching point 114 and is connected via a channel to the feed channel 111 between the first pump device 121 and the third pump device 123. In each case, a valve is arranged between the third pump device 123 and the first pump device 121, between the third pump device 123 and the further chamber 117, between the further chamber 117 and the second pump device 122, and between the second pump device 122 and the branching point 114.

[0078] FIG. 7 shows a flow diagram of an operating method 700 according to one exemplary embodiment. The operating method 700 can be carried out so as to operate the microfluidic apparatus from one of the figures described above or a similar microfluidic apparatus or to control an operation of same.

[0079] The operating method 700 has a step 710 of introducing the sample liquid or a sample into the apparatus. The operating method 700 then involves an effecting step 730 in which conveyance of the sample liquid as first phase, and the sealing liquid as second phase, through the microfluidic network into the division chamber is effected in order to arrange partial volumes of the sample liquid in the cavities and to seal them therein using the sealing liquid. According to the exemplary embodiment illustrated here, the step 730 of effecting conveyance has a production sub-step 732, a transporting sub-step 734 and an introduction sub-step 736, as discussed below.

[0080] In the production sub-step 732, a multi-phase system is produced from the sample liquid as first phase and from at least one further phase, which comprises the sealing liquid and/or a transport liquid, in the microfluidic network. The multi-phase system can for example be realized by embedding the sample liquid or first phase into a second phase which is immiscible or only slightly miscible with the sample liquid and which serves both as sealing liquid and as transport liquid. Alternatively, the sample liquid and the sealing liquid may be embedded on one or both sides into a further, third phase which serves as transport liquid. According to one exemplary embodiment, the liquids used, with the exception of components of the sample liquid, are in particular already pre-stored in the apparatus prior to the introduction step 710.

[0081] In the transporting sub-step 734, the multi-phase system is transported via the feed channel to the channel branching point by means of the at least one pump device. Here, the at least one valve is controlled such that a transport liquid which is optionally present in the multi-phase system is discharged via the discharge channel. In other words, in this case the multi-phase system is microfluidically transported via the supply channel to the channel branching point by means of at least one pump device, wherein a first valve is closed and the transport liquid is discharged via the discharge channel and an open second valve.

[0082] In the introduction sub-step 736, the sample liquid, followed by the sealing liquid, is introduced via the supply channel into the division chamber. Here, the at least one valve is switched over after a boundary interface between the sample liquid and the optionally present transport liquid has passed the channel branching point. In this case, in particular after the boundary interface between sample liquid and transport liquid, which may be identical to the sealing liquid, that is to say which is realized by a liquid having the same physicochemical properties, has passed the channel branching point, the second valve is closed and the first valve opened, with the result that the sample liquid, followed by the sealing liquid, is introduced via the supply channel into the division chamber. In this way, the cavities or compartments of the aliquoting structure are first filled with the sample liquid and then overlaid with the sealing liquid, such that the sample liquid is finally aliquoted in the cavities or compartments.

[0083] According to one exemplary embodiment, the method 700 also has a step 720 of putting the apparatus into a processing unit which is used, inter alia, to control the microfluidic flow within the apparatus. In order to control the microfluidic flow in the apparatus, it is for example possible to produce a pneumatic connection between the apparatus and the processing unit, said pneumatic connection allowing controlled application of pressures to the apparatus. Additionally or alternatively, it is possible to produce a mechanical connection between the apparatus and the processing unit, said mechanical connection making it possible to transmit mechanical forces onto the apparatus, for example for the purpose of releasing liquid reagents pre-stored in the apparatus, and/or making it possible to set the apparatus into controlled rotation, with the result that the liquids enclosed in the apparatus can be processed by means of the inertia forces or pseudo forces, such as centrifugal, Coriolis or Euler forces, resulting from the rotational movement of the apparatus. Additionally or alternatively, the processing unit may have further interfaces to the microfluidic apparatus, which are established in particular in the putting-in step 720, in order to for example at least locally control the temperature of the apparatus and/or detect an optical signal and/or introduce ultrasound and/or introduce mechanical energy and/or couple-in electromagnetic energy.

[0084] According to one exemplary embodiment, after the effecting step 730, the method 700 for operating the microfluidic apparatus also has a step of controlling the temperature, in particular cyclically controlling the temperature, of the division chamber, which contains the cavities or compartments of the aliquoting structure, by means of the temperature-control device or thermal interface or heat-exchange interface. In this way, thermally influenced chemical reactions, for example polymerase chain reactions, can be carried out in the aliquots of the sample liquid which are present in the individual cavities or compartments of the aliquoting structure.

[0085] According to one exemplary embodiment, in a detecting step, a detection device, in particular an optical interface, is additionally used to detect a fluorescence signal which is emitted in particular by the sample liquid in the cavities. It is thus for example possible for the presence of specific deoxyribonucleic acid sequences in the sample liquid to be indicated by using a fluorescent oligonucleotide probe (e.g. TaqMan probe) which is quenched by means of Förster resonance energy transfer (FRET) and which can be cleaved by a polymerase. As result of the use of such fluorescent probes, the course of polymerase reactions in the aliquots of the sample liquid can thus be quantitively monitored in real time. In particular, in this case suitable orientation of the apparatus makes it possible to discharge gas bubbles that form during the temperature-control operation by means of the acting buoyancy force.

[0086] According to one exemplary embodiment, the operating method 700 also has a step of degassing one or more of the liquids, in particular the sealing liquid, for example thermal degassing within the apparatus in a further chamber which has a second temperature-control device or thermal interface. In this way, the quantity of gas bubbles that form during the temperature-control operation in the central chamber can be reduced. In particular, degassing and/or heating of the multi-phase system, in particular of the sample liquid and of the sealing liquid, within the further chamber provided therefor is carried out prior to the transporting sub-step 134, that is to say before the sample liquid and the sealing liquid are successively transported into the division chamber. Optionally, only the sealing liquid is heated and thermally degassed in the further chamber. After the sealing liquid has been degassed in the further chamber, it is pumped, in particular after the introduction sub-step 736 and prior to the temperature-control step, into the division chamber such that the quantity of sealing liquid present in the division chamber is replaced by the quantity of sealing liquid that has previously been heated and thermally degassed in the further chamber. In this way, the quantity of gas bubbles that form in particular during the thermal processing in the temperature-control step in the division chamber can be reduced.

[0087] Exemplary dimensions and specifications of the apparatus 100 are outlined briefly below with reference to the figures described above.

[0088] Lateral dimensions of the apparatus 100 are for example 30×30 mm.sup.2 to 300×300 mm.sup.2, preferably 50×50 mm.sup.2 to 100×100 mm.sup.2. Polymer substrates have a thickness for example of 0.6 mm to 30 mm, preferably 1 mm to 10 mm. A polymer membrane has a thickness for example of 50 μm to 500 μm, preferably 100 μm to 300 μm. Cross sections of the microfluidic channels 111, 112, 113 are for example 100×100 μm.sup.2 to 3×3 mm.sup.2, preferably 300×300 μm.sup.2 to 1×1 mm.sup.2. The pump chambers of the pump devices 121, 122, 123 have a volume for example of 30 nl to 100 μl, preferably 100 nl to 30 μl. Dimensions of the division chamber 115 comprising the aliquoting structure are for example 3×3×0.1 mm.sup.3 to 30×30×3 mm.sup.3, preferably 3×3×0.3 mm.sup.3 to 10×10×1 mm.sup.3. The division chamber 115 comprising the aliquoting structure has a volume for example of ˜1 μl to ˜3 ml, preferably ˜3 μl to ˜100 μl. The cavities or compartments 140 of the aliquoting structure have a volume for example of 10 μl to 10 μl, preferably 10 nl to 300 nl. Lateral dimensions of the temperature-control device or thermal interface 201, 202 are for example 1×1 mm.sup.2 to 100×100 mm.sup.2, preferably 3×3 mm.sup.2 to 30×30 mm.sup.2.

[0089] The sample liquid or first phase 10 comprises, for example, aqueous solutions, in particular for carrying out chemical, biochemical, medical or molecular diagnostic analyses, in particular with sample material, in particular of human origin, e.g. obtained from bodily fluids, smears, secretions, sputum or tissue samples, contained therein. Targets to be detected in the sample liquid have in particular medical, clinical, therapeutic or diagnostic relevance and can for example be bacteria, viruses, specific cells, such as for example circulating tumor cells, cell-free DNA, proteins or other biomarkers.

[0090] The sealing liquid or second phase 20 and the transport liquid or third phase 30 comprise, in particular, mineral oils, silicone oils, fluorinated hydrocarbons, such as for example 3M Fluorinert or Fomblin in suitable combination, wherein the two phases are immiscible or only slightly miscible with one another (for example 3M Fluorinert FC-40 or FC-70 and silicone oil), in particular having a low water solubility in order to prevent undesired mixing with the sample liquid or first phase 10, and/or having a low viscosity in order to obtain a high mobility, i.e. satisfactory discharging of gas bubbles 50 that form, and/or having a low thermal conductivity in order to keep the occurring parasitic heat losses as low as possible, and/or having a low thermal capacity in order to keep the thermal mass to be processed as small as possible, and/or containing surfactants in order to stabilize the boundary interface to the sample liquid or first phase 10.

[0091] The apparatus 100 is in particular primarily manufactured from polymers such as for example polycarbonate (PC), polypropylene (PP), polyethylene (PE), cycloolefin copolymer (COP, COC), polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS) or thermoplastic elastomers (TPE) such as polyurethane (TPU) or styrene block copolymer (TPS), in particular by high-throughput methods such as injection molding, thermoforming, punching, laser transmission welding. Where appropriate, the apparatus 100, in particular in the region of the heat-exchange interface or thermal interface or temperature-control device 201, is provided with components of materials having a high thermal conductivity, such as for example metals such as aluminum, copper, silver or alloys or silicon, in order to obtain an improved exchange of heat between liquids 10, 20, 30 enclosed in the apparatus 100 and the heating and/or cooling apparatuses used.

[0092] The microfluidic pump devices 121, 122, 123 and valves 130, 131, 132 are realized for example by the pneumatically actuated deflection of a polymer membrane into apertures in at least one polymer substrate, in which microfluidic channels and chambers are arranged.

[0093] If an exemplary embodiment comprises an “and/or” combination between a first feature and a second feature, this is to be read as meaning that the exemplary embodiment has, in one embodiment, both the first feature and the second feature and, in a further embodiment, either only the first feature or only the second feature.