Microfluidic device for analysing sample material and method for operating a microfluidic device

Abstract

The invention relates to a microfluidic device (100) for analyzing sample material, wherein the device (100) comprises a microfluidic network (105), the network (105) comprising a first subnetwork (110) for purifying sample material. In the first subnetwork (110), a first microfluidic pump chamber (151) and a second microfluidic pump chamber (152) are connected in series to a functional module (270) for filtering sample material.

Description

State of the art

[0001] The invention relates to a microfluidic device for analyzing sample material and a method for operating a microfluidic device according to the preamble of the independent claims. The present invention also relates to a computer program.

[0002] Microfluidic analysis systems, so-called lab-on-chips (LoCs), enable automated, reliable, fast, compact, and cost-effective processing of patient samples for medical diagnostics. By combining a variety of operations for the controlled manipulation of fluids, complex molecular diagnostic test sequences can be performed in a lab-on-chip cartridge, which can also be referred to as a microfluidic analysis device. Lab-on-chip cartridges can, for example, be manufactured cost-effectively from polymers using series production processes such as injection molding, stamping, or laser transmission welding. The processing of a lab-on-chip cartridge takes place in an associated analysis device. Different types of microfluidic analysis systems are known.

[0003] DE 10 2021 203 638 A1 discloses a corresponding microfluidic device for analyzing sample material, wherein the device comprises a first sub-network for purifying sample material and a second sub-network for amplifying sample material, connected to the first sub-network by a connecting channel. Disclosure of the invention

[0004] Against this background, the approach presented here provides a microfluidic device for analyzing sample material and a method for operating a microfluidic device, as well as a control device using this method and, finally, a corresponding computer program according to the main claims. The measures listed in the dependent claims enable advantageous refinements and improvements of the device specified in the independent claim.

[0005] In general, a pressure-based microfluidic analysis device comprises a multitude of active microfluidic elements, such as valves and pumping chambers, which are interconnected in a suitable network of microfluidic channels. The pneumatic control of the active microfluidic elements is achieved via pneumatic microchannels within the microfluidic device, which can be subjected to positive or negative pressure via a compact interface between the microfluidic analysis device and an analysis device, which can also function as a processing unit. This induces actuation of the microfluidic elements by correspondingly deflecting an elastic membrane.When designing such a microfluidic analysis device, the question arises in particular as to a particularly advantageous arrangement of the elements and design of the microfluidic network in order to be able to address a given range of applications with the microfluidic device in a particularly advantageous manner. The microfluidic device presented here offers a particularly advantageous design of a microfluidic network with active, pneumatically controllable elements for processing a sample fluid. This advantageously minimizes the consumption of reagents required to perform a test procedure, while simultaneously achieving particularly high reliability in microfluidic processing.

[0006] A microfluidic device for analyzing sample material is presented, wherein the device has a microfluidic network, wherein the network comprises a first sub-network for extracting and thus purifying sample material and optionally a second sub-network connected to the first sub-network by a connecting channel for replicating and thus amplifying sample material.

[0007] If the microfluidic device comprises the second subnetwork, the first subnetwork and the second subnetwork can optionally be arranged in a linear topology. Furthermore, the subnetworks can be separable from each other.

[0008] The first subnetwork comprises a first functional module for providing liquid reagents, a second functional module for introducing a sample substance, a third functional module for filtering sample material, a fourth functional module for storing a liquid, a first microfluidic pumping chamber, and a second microfluidic pumping chamber for establishing fluid transport. According to a particular embodiment, the second and fourth functional modules can also be configured as a single functional module. The first microfluidic pumping chamber and the second microfluidic pumping chamber are connected in series with the third functional module for filtering sample material.

[0009] The optional second subnetwork comprises a first functional module for amplifying sample material and optionally a second functional module for dissolving at least one amplification reaction bead.

[0010] The microfluidic device can, for example, be a lab-on-chip cartridge as a component of a pressure-based microfluidic analysis system. The liquid transport within the microfluidic analysis device can occur through the application of pressure, usually pneumatic. The sample material, which can, for example, originate from a sample substance introduced into the microfluidic device and can be processed within the microfluidic device, can, for example, be a sample liquid. A sample substance can be understood to mean the sample that can be introduced into the cartridge, for example a liquid sample or a swab sample. On the other hand, the sample material can also, in particular, be merely components of the sample substance that were obtained from the sample substance, for example by extraction.For example, this sample fluid can be an aqueous solution, for example, obtained from a biological substance, for example of human origin, such as a body fluid, a swab, a secretion, sputum, a tissue sample, or a device with attached sample material. The sample fluid can contain, for example, species of medical, clinical, diagnostic, or therapeutic relevance, such as bacteria, viruses, cells, circulating tumor cells, cell-free DNA, proteins, or other biomarkers, or in particular, components of these objects.For example, the sample fluid can be a master mix or components thereof, for example for carrying out at least one amplification reaction in the microfluidic analysis device, for example for DNA detection at the molecular level, such as an isothermal amplification reaction or a polymerase chain reaction. In this case, sample material can first be extracted in the device from an input sample substance, for example a liquid sample or swab sample. This means that sample material can be purified to enable subsequent amplification of components of the sample substance. Purification is particularly advantageous because the original sample substance can contain components, so-called inhibitors, which would adversely affect the amplification reaction.In the subsequent amplification reaction, only components of the original sample substance, such as specific DNA base sequences, can be amplified, i.e., replicated. By locally applying positive or negative pressure, an elastic membrane, for example, as a component of the microfluidic device, can be specifically deflected into recesses within the microfluidic device, thereby drawing fluid into or expelling fluid from the recesses. The elastic membrane simultaneously creates a separation between the microfluidic, fluid-carrying areas of the device, on the one hand, and the pneumatic areas and the external environment, on the other.Depending on the size of a recess in a fluid-conducting region and the associated displacement volume upon deflection of the membrane into this region, a microfluidic pump chamber with a large displacement volume can be used, for example, to primarily generate a flow in the device, or a microfluidic valve with a small displacement volume can be used to primarily control the flow in the device. By combining at least three microfluidic pump chambers and, additionally or alternatively, valves, a directed flow can be generated in the microfluidic device by successively actuating the elements. For example, a pump chamber can be combined with two surrounding valves, i.e., an inlet and an outlet valve, to achieve directed fluid transport.In addition, for example, three similar elements with comparable displacement volumes can be combined to achieve peristaltic fluid transport through serial, successive actuation.

[0011] For example, the functional modules of the first subnetwork can be used successively to purify or extract sample material. For example, the first functional module can provide liquid reagents in which sample material can be dissolved and additionally or alternatively transported within the device. A sample substance, for example in the form of a sample liquid or a swab sample, can be introduced into the second functional module and, if necessary, removed from a carrier element such as a sampling device. Using the third functional module, for example, certain species can be filtered out of the sample substance so that only the portion of the sample material to be analyzed can be transported further.In the fourth functional module, for example, liquid that arises during the purification of the sample substance can be stored or temporarily stored. The pump module can be used to transport liquid between the different functional modules and additionally or alternatively between the sub-networks. For example, after the sample substance has been purified in the functional modules of the first sub-network, a fluid enriched with sample material can be transported through the connecting channel from the first sub-network to the second sub-network. Here, sample material can be replicated or amplified in the first functional module, for example, while a so-called amplification reaction bead can be provided in a second functional module.Advantageously, this allows the number of rinsing steps required to perform a test sequence within the microfluidic device to be reduced. This allows the time required to perform a test sequence to be shortened.

[0012] According to a further embodiment, the fourth functional module of the first subnetwork and the first functional module of the second subnetwork can be fluidically connected by the connecting channel. For example, liquid resulting from the purification of the sample substance can be stored in the fourth functional module of the first subnetwork. Using the connecting channel, a liquid containing sample material can be transferred to the first functional module of the second subnetwork, and sample material can then be amplified. Advantageously, the sole connection of the two functional modules simplifies the separation of individual functional modules and the subnetworks from one another, and the occurrence of cross-contamination between different process steps, i.e., undesired mixing of different liquid solutions used during the test procedure, can be prevented.

[0013] The device presented here advantageously has a delimitation of the various functionalities that can be provided by the microfluidic device in the form of microfluidically separable sub-networks. A sub-network can be understood as a plurality of interconnected microfluidic elements. As mentioned above, the microfluidic elements can be passive elements such as chambers or actuatable elements such as pump chambers, pumps, or valves. Preferably, a sub-network comprises at least two, preferably more than two, microfluidic elements that are fluidically connected to one another. By two separable sub-networks, it is meant in particular that the two sub-networks are arranged in two spatially separate regions and, in a preferred embodiment, can be separated from one another by a planar cut.Alternatively or additionally, two separable sub-networks can be understood to mean that the two sub-networks are connected to one another by one or more channels, preferably only by one channel, wherein the channels or the channel can preferably be fluidically separated from one another by at least one fluidic separating element such as a valve.

[0014] The detachable subnetworks advantageously enable multiple control of microfluidic elements, which can be assigned to different subnetworks. Furthermore, separating the purification and amplification processes into separate subnetworks allows for a reduction in the number of rinsing steps and prevents unwanted microfluidic crosstalk between different process steps.

[0015] By arranging the first subnetwork and the second subnetwork in a linear topology, i.e., by arranging the subnetworks linearly or in a row, a linear network topology is realized, which allows for particularly low dead volumes that can occur during the execution of a test procedure within the microfluidic analysis device. This advantageously reduces the amount of reagents required to perform the test procedure within the microfluidic device.

[0016] The functional modules of the first subnetwork and, additionally or alternatively, of the second subnetwork can be fluidically separated from one another. For example, each functional module can fulfill a specific task within a purification or analysis process and, for this purpose, have a variable number of individual elements such as valves, pumping, or storage chambers. The individual functional modules can be separated from one another, for example, by means of isolation valves. Furthermore, in a preferred embodiment, at least some of the functional modules can be arranged in a linear topology. The functional modules can be controlled successively during a purification and analysis process, for example, to first extract a species from a sample substance and then enable detection of the species or components thereof in separate areas of the presented microfluidic analysis device.Advantageously, this prevents potential crosstalk, i.e., unwanted mixing of different liquid solutions, which can typically occur due to the multiple use of certain areas of the microfluidic network. This can increase the efficiency of chemical reactions occurring within the microfluidic analysis device and enhance the sensitivity that can be achieved during detection.

[0017] The first microfluidic pumping chamber and the second microfluidic pumping chamber can be used to transport fluid through the third functional module for filtering sample material. Furthermore, the two pumping chambers can be used to transport fluid through other functional modules of the first subnetwork and optionally the second subnetwork and / or to transport fluid from the first subnetwork into the second subnetwork. According to one embodiment, the first subnetwork does not comprise any further pumping chamber or pumping device besides the first and second microfluidic pumping chambers.

[0018] By connecting the two pump chambers in series to the third functional module for filtering sample material, the two pump chambers can be used together to generate a pressure difference between ports of the third functional module.

[0019] For this purpose, the device can have a control port for applying a first pressure to the first microfluidic pumping chamber and a control port for applying a second pressure to the second microfluidic pumping chamber. The first pressure can cause the first microfluidic pumping chamber to expel fluid, and the second pressure can cause suction to the second microfluidic pumping chamber. Combined expulsion and suction can create a large pressure difference across the third functional module. For example, the first pressure can be an overpressure and the second pressure a negative pressure, each relative to an ambient pressure.

[0020] According to one embodiment, a first port of the first microfluidic pump chamber can be connected to the first functional module for providing liquid reagents and to the second functional module for introducing a sample substance. A second port of the first microfluidic pump chamber can be connected to a first port of the third functional module for filtering sample material. A first port of the second microfluidic pump chamber can be connected to the second functional module for introducing a sample substance. A second port of the second microfluidic pump chamber can be connected to a second port of the third functional module for filtering sample material. Depending on the embodiment, the corresponding ports can be connected directly, i.e., without a further interposed element, or, for example, detachably, i.e., via at least one interposed microfluidic valve.

[0021] The third functional module can comprise a filter element that can be detachably connected to the first connection of the third functional module and detachably connected to the second connection of the third functional module. Furthermore, the third functional module can comprise at least one microfluidic valve connected between the first and second connections of the third functional module. This allows the filter element to be bypassed. The filter element can be configured as a correspondingly known filter and can enable the extraction of components from a liquid.

[0022] The second functional module can comprise a sample input chamber for inputting a sample substance. The sample input chamber can be detachably connected to a first port of the second functional module and detachably connected to a second port of the second functional module. The first port of the first microfluidic pump chamber can be connected to the first port of the second functional module, and the first port of the second microfluidic pump chamber can be connected to the second port of the second functional module. Advantageously, the pump chamber can also be used to transport a sample substance input into the sample input chamber through the first subnetwork.

[0023] According to one embodiment, the device can have a common control port for applying pressure, wherein the common control port can be connected to an element of a functional module of the first subnetwork by a first control channel and to an element of a functional module of the second subnetwork by a second control channel. The element can be, for example, a valve or a pump chamber or another microfluidic element of the device. For example, an overpressure or underpressure can be applied to the common control port, which can also be referred to as a port or control port, in order to control the elements of the functional modules of the subnetworks. In this advantageous manner, several active elements, each belonging to different functional modules, can be switched via a common control channel.In other words, multiplexing in the control process is possible. By multiplexing the control of the active microfluidic elements, the number of control ports required to control a given microfluidic network can be advantageously reduced. This allows the costs incurred for the analysis device, in particular the processing unit, to be lowered. Furthermore, by multiplexing the control of the active microfluidic elements with a given number of control ports, a microfluidic network with an increased range of functions can be realized. In this way, the range of applications addressed by such a microfluidic analysis device can be expanded.

[0024] According to a further embodiment, the fourth functional module of the first sub-network can have a first shut-off valve for closing the connecting channel and, additionally or alternatively, the first functional module of the second sub-network can have a second shut-off valve for closing the connecting channel. For example, the first shut-off valve can be kept closed while sample material is being processed within the first sub-network. This has the advantage that an undesired premature penetration of a transfer fluid containing sample material used in the first sub-network into the second sub-network can be prevented. Likewise, after transport to the second sub-network, during which both shut-off valves can be open, closing the second shut-off valve can advantageously prevent a backflow of the transfer fluid containing the sample material into the first sub-network.

[0025] According to a further embodiment, the device can comprise a valve control port for applying pressure, wherein the first shut-off valve and the second shut-off valve can be controlled by means of the valve control port. For example, the first shut-off valve and the second shut-off valve can be closed or opened simultaneously by means of the valve control port to enable or prevent fluid transfer between the first sub-network and the second sub-network of the device. The joint control can advantageously reduce the number of control elements required for processing the device, thus reducing the costs of the system.

[0026] According to a further embodiment, the fourth functional module of the first subnetwork can be configured to dispense sample material. For example, if only purification and extraction of sample material is performed in the device, the sample material can be removed from the microfluidic device after performing an extraction, for example, without transferring it to the second subnetwork. For this purpose, the fourth functional module can, for example, comprise an output device for dispensing the sample material. Advantageously, the device can thus be used variably for different analytical processes.

[0027] According to a further embodiment, the first subnetwork can have an additional fourth functional module for storing a liquid. This allows the range of applications of the microfluidic analysis device to be expanded.

[0028] According to a further embodiment, the device can comprise at least one further control port for applying pressure, wherein the further control port can be connected via a further control channel either to an element of a functional module of the first subnetwork or to an element of a functional module of the second subnetwork. The microfluidic device can, for example, comprise a plurality of control ports, wherein some control ports can be configured to control multiple elements of different functional modules, while other control ports can be used to control a specific element of a functional module. This advantageously enables precise control of all elements of the functional modules.

[0029] In addition, a method for operating a variant of the previously presented microfluidic device is presented, wherein the method comprises a step of extracting sample material using the first sub-network, optionally a step of transferring sample material from the first sub-network to the second sub-network, and optionally a step of amplifying sample material using the second sub-network.

[0030] The extraction step comprises a sub-step of filtering in which a pressure difference is generated at the third functional module by combined squeezing of the first microfluidic pumping chamber and suction with the second microfluidic pumping chamber in order to effect the filtering of sample material.

[0031] According to one embodiment, the extraction step may comprise a sub-step of mixing and, additionally or alternatively, a sub-step of dissolving and, additionally or alternatively, a sub-step of lysing and, additionally or alternatively, a sub-step of rinsing. Additionally or alternatively, the optional transferring step may comprise a sub-step of eluting and, additionally or alternatively, the optional amplifying step may comprise a sub-step of dissolving and, additionally or alternatively, a sub-step of amplifying and, additionally or alternatively, a sub-step of detecting.

[0032] This method can be implemented, for example, in software or hardware or in a mixed form of software and hardware, for example in a control unit.

[0033] The approach presented here further provides a control unit configured to perform, control, or implement the steps of a variant of a method presented here in corresponding devices. This embodiment of the invention in the form of a control unit also allows the problem underlying the invention to be solved quickly and efficiently. For example, the control unit can be used to apply pressures suitable for operating the device to the control ports of the microfluidic device.

[0034] For this purpose, the control unit can have at least one computing unit for processing signals or data, at least one memory unit for storing signals or data, at least one interface to a sensor or an actuator for reading sensor signals from the sensor or for outputting control signals to the actuator, and / or at least one communication interface for reading or outputting data embedded in a communication protocol. The computing unit can be, for example, a signal processor, a microcontroller, or the like, wherein the memory unit can be a flash memory, an EEPROM, or a magnetic storage unit.The communication interface can be designed to read in or output data wirelessly and / or wired, wherein a communication interface that can read in or output wired data can read this data, for example, electrically or optically from a corresponding data transmission line or output it to a corresponding data transmission line.

[0035] In this context, a control unit can be understood as an electrical device that processes sensor signals and outputs control and / or data signals depending on them. The control unit can have an interface that can be implemented in hardware and / or software. In a hardware implementation, the interfaces can, for example, be part of a so-called system ASIC, which contains a wide variety of functions of the control unit. However, it is also possible for the interfaces to be separate integrated circuits or to consist at least partially of discrete components. In a software implementation, the interfaces can be software modules that are present, for example, on a microcontroller alongside other software modules.

[0036] Also advantageous is a computer program product or computer program with program code that can be stored on a machine-readable carrier or storage medium such as a semiconductor memory, a hard disk memory or an optical memory and is used to carry out, implement and / or control the steps of the method according to one of the embodiments described above, in particular when the program product or program is executed on a computer or a device.

[0037] Examples of the approach presented here are shown in the drawings and explained in more detail in the following description. It shows: Fig. 1 a schematic representation of a microfluidic device; Fig. 2 a schematic representation of a microfluidic device; Fig. 3 a schematic representation of a microfluidic device; Fig. 4a schematic representation of a microfluidic device; Fig. 5 a schematic representation of an embodiment of a microfluidic device; Fig. 6 a schematic representation of an embodiment of a microfluidic device; Fig. 7 a schematic representation of an embodiment of a microfluidic device; Fig. 8 a schematic representation of an embodiment of a microfluidic device; Fig. 9 a perspective top view of an embodiment of a microfluidic device; Fig. 10 a top view of an embodiment of a microfluidic device; Fig. 11 a perspective top view of a section of the microfluidic device according to an embodiment; Fig. 12 a perspective bottom view of a section of the microfluidic device according to an embodiment; Fig. 13a flowchart of a method 1000 for operating a microfluidic device according to an embodiment; Fig. 14 a flowchart of a method 1000 for operating a microfluidic device according to an embodiment; Fig. 15 a flowchart of a method 1000 for operating a microfluidic device according to an embodiment; Fig. 16 a flowchart of a method 1000 for operating a microfluidic device according to an embodiment; and Fig. 17 a schematic representation of an embodiment of an analytical device for accommodating a microfluidic device.

[0038] In the following description of advantageous embodiments of the present invention, the same or similar reference numerals are used for the elements shown in the various figures and having a similar effect, whereby a repeated description of these elements is omitted.

[0039] Fig. 1 shows a schematic representation of a microfluidic device 100. In this exemplary embodiment, the device 100 comprises a microfluidic network 105, which optionally has a linear topology. The network 105 is divided into a first subnetwork 110 and an optional second subnetwork 120, with both subnetworks 110 and 120 being marked by dashed lines in the illustration shown here.

[0040] Even if embodiments of the microfluidic device 100 with the two sub-networks 110, 120 are shown in this and the following figures, the microfluidic device shown in each case can also be designed with only the first sub-network 110, i.e. can either comprise no second sub-network 120 or comprise a differently designed second sub-network.

[0041] If a corresponding microfluidic device, such as the one in Fig. 1While the microfluidic device 100 shown comprises both the first subnetwork 110 and the second subnetwork 120, the two subnetworks 110, 120 are connected to each other via a microfluidic connecting channel 125. Optionally, the first subnetwork 110 and the second subnetwork 120 are arranged in a linear topology. However, according to this and the embodiments illustrated in the following figures, a different arrangement of the subnetworks 110, 120 is also possible.

[0042] Both the first subnetwork 110 and the second subnetwork 120 comprise further microfluidic channels, which are schematically represented in the figure shown here as solid lines, which connect different microfluidic elements to each other. In this exemplary embodiment, the network 105 of the microfluidic device 100 is characterized in particular by twenty active microfluidic valves 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150. For example only, two valves 144, 145 are arranged on the connecting channel 125 and are designed as a first shut-off valve 144 and a second shut-off valve 145 for closing the connecting channel. Thus, the subnetworks 110, 120 are separable from one another.The network 105 also comprises six active microfluidic pump chambers 151, 152, 153, 154, 155, 156, a passive microfluidic chamber 160, three pre-storage chambers with vent openings 161, 162, 163 for long-term stable pre-storage of liquid reagents, two sample input chambers 167, 168 which enable sample input, a filter element 170 which enables extraction of components from a liquid, a liquid storage chamber 180 which serves to receive liquids after processing in the microfluidic network 105, and four venting devices 191, 192, 193, 194 which serve to vent the microfluidic network. In this embodiment, the total of twenty-six active microfluidic elements can be controlled by pneumatic control channels, as illustrated by the lines in the reference symbols branching off perpendicularly to the microfluidic channels.The arrangement and linking of the named elements forms the microfluidic network 105, which has a linear topology only as an example.

[0043] The microfluidic network 105 of the microfluidic device 100 is composed of the first subnetwork 110, which is intended for the purification of sample material, and the second subnetwork 120, which is intended for the amplification of sample material. The two subnetworks 110, 120 are fluidically connected to one another via a microfluidic connecting channel 125. This enables microfluidic processing within the two subnetworks 110, 120 to be carried out successively, i.e., one after the other, within the scope of a test sequence. Accordingly, a combination of active microfluidic elements, each arranged in different subnetworks 110, 120, can be controlled, if necessary, via a common control channel.

[0044] Fig. 2shows a schematic representation of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figure and comprises a microfluidic network 105, which is divided into a first sub-network 110 and a second sub-network 120. In this exemplary embodiment, the microfluidic elements of the network 105 are arranged within the sub-networks 110, 120 in a plurality of separable functional modules, wherein the individual functional modules are each marked in the illustration shown here by framing rectangular boxes made of dot-dashed lines. In detail, the sub-network 110 for purifying sample material in this exemplary embodiment comprises a pump module 230 for transporting fluids and sample material dissolved therein within the network 105 or between the different functional modules.For example only, the pump module comprises two active microfluidic pump chambers 151, 152 and a microfluidic valve 136. Furthermore, the first subnetwork 110 has a first functional module 250 for providing liquid reagents. In this first functional module, three pre-storage chambers 161, 162, 163 are arranged for long-term stable pre-storage of liquid reagents. For example only, the liquid reagents are aqueous solutions, for example, buffer solutions for processing a sample substance or components thereof. In another embodiment, mineral oils, silicone oils, or fluorinated hydrocarbons can also be stored in the pre-storage chambers.In this exemplary embodiment, each pre-storage chamber 161, 162, 163 is fluidically connected to a respective microfluidic valve 131, 132, 133, which in turn are fluidically connected to one another via a first channel interface 255 and to a second channel interface 257. By closing the valves 131, 132, 133, the first functional module 250 can be separated from the remaining microfluidic network 105. Via the second channel interface 257, the first functional module 250 in this exemplary embodiment is connected to the pump module 230, to a second functional module 261 for introducing sample substances, and, merely by way of example, to an additional second functional module 262 for introducing sample substances. The second functional module 261 comprises a sample introduction chamber 167 with a venting device 191.The sample input chamber 167 is designed merely by way of example to receive a swab sample, i.e., a sampling device with attached sample material, and is fluidically connected to two valves 134, 135 within the second functional module. Similarly, the additional second functional module 262 comprises a sample input chamber 168 with a venting device 192, which is fluidically connected to two valves 137, 138. In this exemplary embodiment, the sample input chamber 168 is designed merely by way of example to input a liquid sample. The second functional module 261 and the additional second functional module 262 are fluidically connected to the second channel interface via the valves 134, 137.Via the valves 135, 138, the second functional module 261 and the additional second functional module 262 are in turn fluidically connected to a third channel interface 265, via which, merely by way of example, a fluidic connection exists to the pump module 230 and to a third functional module 270 for filtering the sample material. By closing the valves 134, 135, the second functional module 261 can be separated from the remaining microfluidic network 105, just as the additional second functional module 262 can be separated from the remaining microfluidic network 105 by closing the valves 137, 138. In this embodiment, the third functional module 270 comprises, in addition to four microfluidic valves 139, 140, 141, 142, a filter element 170, which is merely an example of a silica filter and is designed merely as an example to filter out species from a sample substance.Closing the valves 139, 140, which are fluidically connected to the third channel interface 265 and a fourth functional module 280, directs a fluid flow through the filter element 170. By closing the valves 139, 141, the third functional module 270 can be separated from the functional modules 250, 261, 262 and the pump module 230, and by closing the valves 140, 142, the third functional module 270 can be separated from the fourth functional module 280. In this exemplary embodiment, the fourth functional module 280 is designed to store a fluid used in the network 105. For this purpose, the fourth functional module 280 has, merely as an example, a liquid storage chamber 180 with a venting device 193 and two valves 143, 144. In another embodiment, the fourth functional module 280 may additionally be configured to dispense sample material.In this exemplary embodiment, the fourth functional module 280 is fluidically connected to the third functional module 270 and the connecting channel 125 via a fourth channel interface 285. The valve 144 is designed, merely by way of example, as a shut-off valve and is arranged on the connecting channel 125. By closing the valve 144, the entire first subnetwork 110 can thus be separated from the remaining microfluidic network 105. In this exemplary embodiment, the connecting channel 125 represents a fluidic connection between the fourth functional module 280 of the first subnetwork 110 and a first functional module 290 of the second subnetwork 120 for amplifying sample material.The first functional module 290 of the second subnetwork 120 is designed in this embodiment to amplify sample material and, for this purpose, comprises, by way of example only, three active microfluidic pump chambers 153, 154, 155 arranged in a row, which are bracketed by two microfluidic valves 145, 146, by way of example only. In this embodiment, the valve 145 is arranged directly on the connecting channel 125 and is designed as a shut-off valve. By closing the valve 145, the entire second subnetwork 120 can thus be separated from the remaining microfluidic network 105. In this embodiment, the second subnetwork 120 also has a second functional module 295 with four valves 147, 148, 149, 150, an active microfluidic pump chamber 156, a passive microfluidic chamber 160 and a venting device 194.The first functional module 290 and the second functional module 295 are fluidically connected to one another via the valves 146, 147. Separation of the two functional modules 290, 295 from one another is thus possible both by means of the valve 146 of the first functional module 290 and by means of the valve 147 of the second functional module 295. The second functional module 295 of the second subnetwork 120 is designed, merely by way of example, to provide at least one amplification reaction bead. The amplification reaction bead is, merely by way of example, a lyophilized or freeze-dried reagent for producing an amplification reaction mix.

[0045] Fig. 3shows a schematic representation of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures. In the illustration shown here, the functional modules 250, 261, 262, 270, 280, 290, 295 and the pump module 230 are shown in a simplified manner without the individual microfluidic elements. The functional modules 250, 261, 262, 270, 280, 290, 295 and the pump module 230 are arranged in a linear topology. Furthermore, the functional modules 250, 261, 262, 270, 280 and the pump module 230 are designed as components of the first sub-network 110 and the functional modules 290, 295 as components of the second sub-network 120, wherein the first sub-network 110 and the second sub-network 120 are connected to one another via exactly one connecting channel 125.The selected topology makes it possible, for example, to reduce the size of dead volumes, reduce the number of pumping steps and prevent unwanted microfluidic crosstalk between different process steps of a sequence.

[0046] Fig. 4shows a schematic representation of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures with the described network. In addition to the microfluidic elements that form the network 105 of the microfluidic device 100, the control channels that serve to control the active microfluidic valves 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150 and pump chambers 151, 152, 153, 154, 155, 156 are also shown as dashed lines. In this particularly advantageous embodiment, these twenty-six active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156 by means of a compact control interface 400. The control interface 400 comprises, by way of example, only twenty control connections 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, which can also be referred to as control ports. In this embodiment, the control ports are pneumatic connections, via which the active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156 are actuated, merely by applying positive or negative pressure. This achieves a total of six-fold multiplexing, as results from the difference between the number of twenty-six elements and twenty ports. In the figure shown here, the associated six branches of pneumatic control channels that enable the multiplexing are shown.illustrated in the form of black nodes. The branching of the control channels occurs either directly at a control port or at a suitable point on a control channel. In the exemplary embodiment shown, a branching occurs directly at the control connection 409 merely as an example. The control connection 409 is designed merely as an example to jointly control the valve 141 of the third functional module 270 of the first sub-network 110 through a first control channel 431 and the valve 148 of the second functional module 295 of the second sub-network 120 through a second control channel 432. Similarly, in this exemplary embodiment, the first shut-off valve 144 and the second shut-off valve 145 are controllable by means of a valve control connection 418 for applying pressure. This makes it possible to open or close the connecting channel 125 using the valve control connection 418, and the individual sub-networks 110,120 are separable from each other. In contrast, a second control port 410 in this exemplary embodiment has a first branch 441 and a second branch 442 and is thus designed to jointly control both the valve 131 of the first functional module 250 of the first sub-network 110 and the valves 149, 150 of the second functional module 295 of the second sub-network 120. The control interface 400 also comprises a further control port 401 for applying pressure, wherein the further control port 401 in this exemplary embodiment is connected to the valve 132 of the first functional module 250 of the first sub-network 110 via a further control channel 451. For example only, a second further control port 402 is connected to the pump chamber 152 of the pump module 230 of the first sub-network 110.a third further control connection 403 is connected to the valve 137 of the additional second functional module 262 of the first sub-network 110, a fourth further control connection 404 is connected to the valve 138 of the additional second functional module 262 of the first sub-network 110, a fifth further control connection 405 is connected to the valves 139, 140 of the third functional module 270 of the first sub-network 110, a sixth further control connection 406 is connected to the valve 143 of the fourth functional module 280 of the first sub-network 110, an eighth further control connection 408 is connected to the valve 142 of the third functional module 270 of the first sub-network 110, an eleventh further control connection 411 is connected to the valve 133 of the first functional module 250 of the first sub-network 110, a twelfth further control connection 412 is connected to the valves 134,135 of the second functional module 261 of the first subnetwork 110, a thirteenth further control connection 413 is connected to the pump chamber 151 of the pump module 230 of the first subnetwork 110, and a fourteenth control connection 414 is connected to the valve 136 of the pump module 230 of the first subnetwork 110. Likewise, elements of the second subnetwork 120 of the device 100 in this exemplary embodiment can also be controlled by means of the control interface 400. For example, a seventh further control connection 407 is connected to the pump chamber 153 of the first functional module 290 of the second sub-network 120, a fifteenth further control connection 415 is connected to the valve 146 of the first functional module 290 of the second sub-network 120, a sixteenth further control connection 416 is connected to the pump chamber 155 of the first functional module 290 of the second sub-network 120,a seventeenth further control connection 417 is connected to the pump chamber 154 of the first functional module 290 of the second sub-network 120, a nineteenth further control connection 419 is connected to the valve 147 of the second functional module 295 of the second sub-network 120, and a twentieth further control connection 420 is connected to the pump chamber 156 of the second functional module 295 of the second sub-network 120.

[0047] Fig. 5 shows a schematic representation of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures.

[0048] Optional is in contrast to the one based on Fig. 3 described device, instead of an additional second functional module for inputting sample substances, an additional fourth functional module 580 for storing a liquid is shown.

[0049] In contrast to the one based on Fig. 3 According to this embodiment, a first microfluidic pumping chamber 151 and a second microfluidic pumping chamber 152 are used to transport liquid between the functional modules 250, 261, 270, 280, 290, 295, 580.

[0050] In the figure shown here, the functional modules 250, 261, 270, 280, 290, 295, and 580 are shown in a simplified manner without the individual microfluidic elements. The functional modules 250, 261, 270, 280, 290, 295, and 580 are arranged in a linear topology. The functional modules 250, 261, 270, 280, and 580 as well as the pump chambers 151, 152 are designed as components of the first subnetwork 110, and the functional modules 290, 295 are designed as components of the second subnetwork 120, with the first subnetwork 110 and the second subnetwork 120 being connected to one another via precisely one connecting channel 125.

[0051] The first microfluidic pump chamber 151 is connected between the first functional module 250 for providing liquid reagents and the third functional module 270 for filtering sample material. For example, a first port of the first microfluidic pump chamber 151 is coupled to a port of the first functional module 250, a first port of the second functional module 261, and a first port of the additional fourth functional module 580. A second port of the first microfluidic pump chamber 151 is coupled, for example, to a first port of the third functional module 270 and a second port of the additional fourth functional module 580. Thus, the first microfluidic pump chamber 151 is connected in parallel to the optional additional fourth functional module 580.

[0052] The second microfluidic pump chamber 152 is connected between the second functional module 261 and a port of the first subnetwork 110 coupled to the connecting channel 125. For example, a first port of the second microfluidic pump chamber 152 is coupled to a second port of the second functional module 261. A second port of the second microfluidic pump chamber 152 is connected, for example, to a port of the fourth functional module 280 for storing a liquid.

[0053] In this way, a series connection between the microfluidic pump chambers 151, 152 and the third functional module 270 is possible, which makes it possible to operate the microfluidic pump chambers 151, 152 such that they jointly create a pressure difference between the connections of the third functional module 270. For example, the first microfluidic pump chamber 151 can be operated such that the first microfluidic pump chamber 151 creates an overpressure at the first connection of the third functional module 270. At the same time, the second microfluidic pump chamber 152 can be operated such that the second microfluidic pump chamber 152 creates a negative pressure at the second connection of the third functional module 270.

[0054] Fig. 6shows a schematic representation of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figure and comprises a microfluidic network 105, which is divided into a first subnetwork 110 and a second subnetwork 120. In this embodiment, the microfluidic elements of the network 105 are arranged within the subnetworks 110, 120 in a plurality of detachable functional modules, wherein the individual functional modules are each marked in the representation shown here by framing rectangular boxes made of dot-dashed lines.

[0055] The microfluidic device 100 essentially corresponds to the one described in Fig. 2 described device, the essential difference being the arrangement of the microfluidic pumping chambers 151, 152.

[0056] Optional is in contrast to the one based on Fig. 2 In the device described above, instead of an additional second functional module for introducing sample substances, an additional fourth functional module 580 for storing a liquid is shown. The additional fourth functional module 580 comprises an additional liquid storage chamber 680, a venting device 192, and, for example, three valves 137, 138, 638.

[0057] Specifically, in this exemplary embodiment, the subnetwork 110 for purifying sample material comprises a pump module for transporting fluids and sample material dissolved therein within the network 105 or between the different functional modules. For example only, the pump module for this purpose comprises the two active microfluidic pump chambers 151, 152 and the microfluidic valves 136, 151.

[0058] In addition, the first subnetwork 110 has a first functional module 250 for providing liquid reagents. In this first functional module, three pre-storage chambers 161, 162, 163 are arranged, merely by way of example, for the long-term stable pre-storage of liquid reagents. The liquid reagents are aqueous solutions, for example buffer solutions for processing a sample substance or components thereof. In another exemplary embodiment, mineral oils, silicone oils, or fluorinated hydrocarbons can also be stored in the pre-storage chambers. In this exemplary embodiment, each pre-storage chamber 161, 162, 163 is fluidically connected to a respective microfluidic valve 131, 132, 133, which in turn are fluidically connected to one another via a first channel interface 255 and to a second channel interface 257.By closing the valves 131, 132, 133, the first functional module 250 can be separated from the remaining microfluidic network 105.

[0059] In this exemplary embodiment, the first functional module 250 is connected via the second channel interface 257 to the microfluidic valve 651, which is connected in series with the microfluidic pump chamber 151, to a second functional module 261 for introducing sample substances, and, merely by way of example, to the additional fourth functional module 580 for introducing sample substances. A first connection of the first pump chamber 151 is connected to the second channel interface 257 via the valve 651.

[0060] The second functional module 261 comprises a sample input chamber 167 with a venting device 191. The sample input chamber 167 is designed merely as an example to receive a swab sample, i.e. a sampling device with attached sample material, and is fluidically connected to two valves 134, 135 within the second functional module 261.

[0061] The second functional module 261 and the additional fourth functional module 580 are fluidically connected to the second channel interface 257 via the valves 134, 137. The additional fourth functional module 580 is in turn fluidically connected to a third channel interface 165 via the valves 138, 138, which, merely by way of example, provides a fluidic connection to the first pump chamber 151 and to a third functional module 270 for filtering the sample material. By closing the valves 134, 135, the second functional module 261 can be separated from the remaining microfluidic network 105, just as the additional fourth functional module 262 can be separated from the remaining microfluidic network 105 by closing the valves 137, 138, 138.

[0062] In this exemplary embodiment, the third functional module 270 comprises, in addition to four microfluidic valves 139, 140, 141, 142, a filter element 170, which is merely an example of a silica filter and is designed merely as an example to filter species from a sample substance. Closing the series-connected valves 139, 140, which are connected between the third channel interface 265 and a fourth channel interface 285, directs a fluid flow through the filter element 170. By closing the valves 139, 141, the third functional module 270 can be separated from the functional modules 250, 261, 262 and the pump module 230, and by closing the valves 140, 142, the third functional module 270 can be separated from the fourth functional module 280.

[0063] In this exemplary embodiment, the fourth functional module 280 is designed to store a fluid used in the network 105. For this purpose, the fourth functional module 280 has, merely as an example, a liquid storage chamber 180 with a venting device 193 and two valves 143, 144. In another exemplary embodiment, the fourth functional module 280 can also be designed to dispense sample material. In this exemplary embodiment, the fourth functional module 280 is fluidically connected to the fourth channel interface 285 via the second pump chamber 252 and the valve 136.

[0064] Thus, the third functional module 270 for filtering sample material is connected between the third channel interface 265 and the fourth channel interface 285. A second connection of the first pump chamber 151 is connected directly to the third channel interface 265, and a second connection of the second pump chamber 152 is connected to the third channel interface 265 via the valve 136.

[0065] The fourth channel interface 285 is connected to the connecting channel 125 via the valve 144, which is designed merely as a shut-off valve by way of example. By closing the valve 144, the entire first subnetwork 110 can thus be separated from the remaining microfluidic network 105 and in particular from the second subnetwork 120. In this exemplary embodiment, the connecting channel 125 represents a fluidic connection between the fourth functional module 280 of the first subnetwork 110 and a first functional module 290 of the second subnetwork 120 for amplifying sample material.

[0066] The first functional module 290 of the second subnetwork 120 is designed in this embodiment to amplify sample material and, for this purpose, comprises, by way of example only, three active microfluidic pump chambers 153, 154, 155 arranged in a row, which are bracketed by two microfluidic valves 145, 146, by way of example only. In this embodiment, the valve 145 is arranged directly on the connecting channel 125 and is designed as a shut-off valve. By closing the valve 145, the entire second subnetwork 120 can thus be separated from the remaining microfluidic network 105. In this embodiment, the second subnetwork 120 also has a second functional module 295 with four valves 147, 148, 149, 150, an active microfluidic pump chamber 156, a passive microfluidic chamber 160 and a venting device 194.The first functional module 290 and the second functional module 295 are fluidically connected to one another via the valves 146, 147. Separation of the two functional modules 290, 295 from one another is thus possible both by means of the valve 146 of the first functional module 290 and by means of the valve 147 of the second functional module 295. The second functional module 295 of the second subnetwork 120 is designed, merely by way of example, to provide at least one amplification reaction bead. The amplification reaction bead is, merely by way of example, a lyophilized or freeze-dried reagent for producing an amplification reaction mix.

[0067] Figure 7shows a schematic representation of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures with the described network. In addition to the microfluidic elements that form the network 105 of the microfluidic device 100, the control channels that serve to control the active microfluidic valves 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156 are also shown as dashed lines. In this embodiment, these active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 638, 651 are controlled by means of a compact control interface 400.The control interface 400 comprises, by way of example, only twenty control connections 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, which can also be referred to as control ports. In this exemplary embodiment, the control ports are pneumatic connections via which the active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 638, 651 are actuated, merely by applying positive or negative pressure. This achieves multiple multiplexing. In the embodiment shown here. Fig. 7The corresponding branches of pneumatic control channels, which enable multiplexing, are illustrated as black nodes. The control channels are branched either directly at a control port or at a suitable point on a control channel.

[0068] Thus, Fig. 7 a schematic representation of the network 105 of a further advantageous embodiment of the microfluidic analysis device 100 with a linear topology.

[0069] Comparable to the previous embodiments, the network 105 of the further embodiment of the microfluidic analysis device 100 is composed of microfluidic channels, which are schematically represented as solid lines, which connect different microfluidic elements to one another. The network 105 of the microfluidic analysis device 100 is characterized in this further embodiment in particular by twenty-two active microfluidic valves 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 638, 651, six active microfluidic pump chambers 151, 152, 153, 154, 155, 156, one passive microfluidic chamber 160, three pre-storage chambers 161, 162, 163 with vent openings for long-term stable pre-storage of liquid reagents, a chamber 167 which enables sample introduction, a filter element 170 which enables extraction of components from a liquid, two liquid storage chambers 180, 680, which serve to hold liquids after processing in the microfluidic network 105, and three venting devices 191, 192, 193 which serve to vent the microfluidic network 105.

[0070] The total of twenty-eight active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 638, 651 can be controlled in this embodiment by pneumatic control channels, which are in Fig. 6 are each illustrated by the lines branching off perpendicularly to the microfluidic channels. A detailed circuit diagram of this advantageous embodiment of the microfluidic analysis device 100, including the design of the pneumatic control channels, is shown in Fig. 7 given.

[0071] As in the previous embodiments, the arrangement and connection of the aforementioned elements to form the microfluidic network 105 is selected with a particularly advantageous, linear topology. As shown in the figures, the microfluidic network 105 of the microfluidic analysis device 100 is composed of a first subnetwork 110, which is intended for the purification of sample material, and a second subnetwork 120, which is intended for the amplification of sample material. The two subnetworks 110, 120 are connected to each other via the microfluidic connecting channel 210.Because the microfluidic processing takes place successively, i.e. one after the other, within the scope of carrying out a test sequence within the two sub-networks 110, 120, a combination of active microfluidic elements, each belonging to a different sub-network 110, 120, can be controlled by a common control channel if necessary, without this resulting in a disadvantage for the microfluidic processing.

[0072] In the Fig. 7sketched circuit diagram of this advantageous embodiment of the microfluidic device 100, it can be seen that in this embodiment, in a particularly advantageous manner, for controlling the total of twenty-eight active microfluidic elements 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 638, 651 only twenty pneumatic control ports 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720 can be used, whereby this can be achieved via a total of eight branches of the pneumatic control channels. The branching points of the pneumatic control channels are in Fig. 7Each is schematically illustrated by black, circular nodes on the pneumatic control channels shown as dashed lines. This exemplary embodiment thus illustrates that the selected inventive design of the microfluidic device 100 and the multiplexing in the control of the elements can provide a significant functional advantage, since, on the one hand, only a reduced number of pneumatic control ports is required to actuate the microfluidic elements and, on the other hand, due to the linear topology of the microfluidic network 105, no significant restrictions in the microfluidic process control have to be accepted. Thus, the particularly advantageous design according to the invention can combine improved functionality with reduced effort for controlling the active microfluidic elements.Furthermore, separating the purification and amplification processes in the separate sub-networks 110, 120 advantageously enables a reduction of dead volumes, a reduction in the number of rinsing steps, and the prevention of undesired microfluidic crosstalk between different process steps.

[0073] Through the Figures 6 and 7 In the advantageous embodiment of the microfluidic device 100 outlined above, a particularly variable pumping strategy can be selected for the purification of a sample substance during the extraction of components of the sample substance by the filter element 170.

[0074] Compared to the Figures 1 to 4 The first embodiment outlined in the drawings is Figures 6 and 7In the further embodiment shown, the two pump chambers 151 and 152 are integrated into the microfluidic network 105 in such a way that they are connected in series with the filter element 170. This enables, for example, improved pumping of a sample liquid via the filter element 170 by combining squeezing of the pump chamber 151 with suction with the pump chamber 152, so that a particularly high pressure difference exists in the microfluidic system when pumping a sample liquid via the filter element 170. In this way, for example, particularly highly viscous or viscous sample substances or inhomogeneous sample liquids with highly viscous sample components can be reliably pumped through the filter element 170 in an advantageous manner, for example to achieve an extraction or accumulation of sample components on the filter element 170.

[0075] Furthermore, a microfluidic separation of the first pump chamber 151 from the pre-storage chambers 161, 162, 163 and the sample input chamber 167 is implemented via at least two of the microfluidic valves 131, 132, 133, 134, 651, so that intrinsically a particularly reliable microfluidic separation of these fluid paths can be ensured in the event of pressurization of the microfluidic pump chamber 151.

[0076] Furthermore, the detailed presentation in Fig. 7 the individual functional modules 250, 261, 270, 280, 290, 295, 580, which form the sub-networks 110, 120 of the microfluidic analysis device 100, each shown by framing rectangular boxes: In detail, the sub-network 110 for the purification of sample material is composed of the first functional module 250 for providing liquid reagents, the second functional module 261 for introducing a sample substance into the microfluidic device 100, the third functional module 270 for filtering out species from the sample substance, two fourth functional modules 280, 580 for storing liquids, and two pump modules 730, 731 for establishing the liquid transport between the different functional modules.

[0077] The subnetwork 120 for amplification of sample material consists of the first functional module 290 for amplification, and the second functional module 295 for providing an amplification reaction bead.

[0078] The Figures 5 to 7show in varying degrees of detail the arrangement of the functional modules 250, 261, 270, 280, 580 in the first sub-network 110 for the purification of sample material and the functional modules 290, 295 in the second sub-network 120 for the amplification of sample material in a linear topology in the further embodiment.

[0079] As described above, in this advantageous embodiment, improved pumping of a sample liquid via the filter element 170 can be enabled by using the two pump modules 151, 152, which are connected on both sides to the third functional module 270 for filtering out species from the sample substance, in the first sub-network 110 for purifying sample material.

[0080] For example, the control port 718 is used to apply a first pressure to the first microfluidic pump chamber 151, and the control port 709 is used to apply a second pressure to the second microfluidic pump chamber 152. Thus, the two pump chambers 151, 152 can be controlled independently of each other.

[0081] The common control port 703 is used, for example, to control the valves 138, 141 of the first subnetwork 110 via a first control channel 431 and the valve 148 of the second subnetwork 120 via a second control channel 432. Accordingly, the common control port 701 is used both to control the first functional module 250 of the first subnetwork 110 and to control the second functional module 295 of the second subnetwork 120.

[0082] The valve control connection 713 is used to control the shut-off valves 144, 145.

[0083] Fig. 8shows a schematic representation of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures. For illustrative purposes, the microfluidic network 105 is shown in a simplified manner in the figure shown here. In this exemplary embodiment, the microfluidic device 100 comprises two sub-networks 110, 120, which are connected to one another via a microfluidic connecting channel 125, and a pneumatic control interface 400 with four control connections 409, 410, 418, 500 and 701, 703, 713, 500, respectively. The control connections 409, 410, 418, 500, 701, 703, 713, 500 are each designed in this exemplary embodiment to control microfluidic elements of the first sub-network 110 as well as the second sub-network 120.Through this quadruple multiplexing, a total of eight active microfluidic elements of the device 100 can advantageously be controlled by means of the four control ports, four of them per subnetwork 110, 120. The multiplexing is achieved by branching the control channels at a total of four nodes 409, 418, 441, 505 and 703, 713, 441, 505.

[0084] Fig. 9shows a perspective top view of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the previous figures. In this embodiment, the device 100 has a reagent bar 600 arranged within the three pre-storage chambers 161, 162, 163 with three compartments, in each of which different liquid reagents can be pre-stored with long-term stability. Accordingly, the pre-storage chambers 161, 162, 163 can also be referred to as reagent pre-storage chambers. In addition, within the sample input chamber 167 of the device described in the previous figures Figures 2 to 4 described second functional module, a sampling device 605 is arranged merely by way of example, which allows the transfer of a swab sample located thereon into the network 105 of the microfluidic device 100.

[0085] Fig. 10shows a top view of an embodiment of a microfluidic device 100. The device 100 shown here corresponds to or is similar to the device described in the preceding figures. In the illustration shown here, some of the elements of the microfluidic device 100 described in the preceding figures are numbered by way of example. Specifically, the microfluidic valve 146, the microfluidic pump chambers 151, 152, 153, 154, 155, 156, the microfluidic chamber 160, the three pre-storage chambers 161, 162, 163, the two sample input chambers 167, 168, the microfluidic filter element 170, and the liquid storage chamber 180 are numbered.Furthermore, the first sub-network 110 for the purification of a sample substance, the second sub-network 120 for the amplification of sample material and the pneumatic control interface 400 with an additional control connection 401 numbered by way of example are shown in dashed lines. For example only, the device has an overall lateral dimension of 118 × 78 mm 2<. In another exemplary embodiment, the overall dimension can be 50 × 25 mm 2< to 300 × 200 mm 2<, preferably 75 × 25 mm 2< to 200 × 100 mm 2<. In this exemplary embodiment, the fluidic and pneumatic microchannels also have a cross-sectional dimension of, for example, 600 × 400 µm 2<. In another embodiment, the cross-sectional dimensions of the fluidic and pneumatic microchannels can be 100 × 100 µm 2< to 3 × 3 mm 2<, preferably 300 × 300 µm 2< to 1 × 1 mm 2<.The effective displacement volume of a microfluidic pump chamber in this embodiment is 20 µl, the effective displacement volume of a microfluidic valve is merely 125 nl by way of example, and the volume of the microfluidic chamber with filter element in this embodiment is merely 8 µl by way of example. In other embodiments, the effective displacement volume of a microfluidic pump chamber can be 1 µl to 50 µl, preferably 5 µl to 30 µl, the effective displacement volume of a microfluidic valve can be 50 nl to 1 µl, preferably 100 nl to 300 nl, and the volume of the microfluidic chamber with filter element can be 3 µl to 20 µl, preferably 5 µl to 10 µl.For microfluidic processing within the device 100 in this embodiment, a pressure difference (overpressure or negative pressure relative to atmospheric pressure), which is used to generate the microfluidic flow by extruding or sucking in a pump chamber or controlling the microfluidic flow by means of a valve, of only 700 mbar, for example, can be used. In another embodiment, the pressure difference can be 100 mbar to 2000 mbar, preferably 400 mbar to 1500 mbar.

[0086] Fig. 11shows a perspective top view of a section of the microfluidic device 100 according to an embodiment. The device 100 shown here corresponds to or is similar to the device described in the previous figures. In the figure shown here, the microfluidic valve 131, the microfluidic pump chambers 151, 152, the sample input chamber 168, the microfluidic chamber with the filter element 170, the venting device 193, and the pneumatic control ports 405, 409, 410 are numbered by way of example. In this embodiment, the microfluidic device 100 is realized by two rigid polymer components, 800, 805, which are connected to one another via a flexible membrane 810. The lower polymer component 800 is closed off by a further polymer film in order to seal the microchannels present in the polymer component.The multilayer structure thus enables, in particular, the realization of fluidic and pneumatic microchannels on at least two different lateral planes. In this way, fluidic and pneumatic microchannels can be crossed, and a particularly high degree of integration of the microfluidic device 100 with a central and particularly compact design of the microchannels shown in FIG. Figure 4This can be achieved using the pneumatic control interface described above. By applying positive or negative pressure to the pneumatic control ports, the flexible polymer membrane can be deflected via the pneumatic microchannels into recesses located on the microfluidic valves and pump chambers, thereby opening or closing a microfluidic valve or pump chamber. By sucking in or expelling fluid via a pump chamber, the microfluidic flow within the network 105 can be precisely controlled with a suitable valve position.In other embodiments, the following materials, for example, can be used to realize the microfluidic analysis device: Primarily polymers such as polycarbonate (PC), polystyrene (PS), styrene-acrylonitrile copolymer (SAN), 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). The device can be manufactured, for example, using series production processes such as injection molding, injection-compression molding, thermoforming, punching, or laser transmission welding.

[0087] Fig. 12shows a perspective bottom view of a section of the microfluidic device 100 according to an embodiment. The device 100 shown here corresponds to or is similar to the device described in the previous figures. In the figure shown here, the microfluidic valve 131, the microfluidic pump chambers 151, 152, the sample input chambers 167, 168, the microfluidic chamber with the filter element 170, the venting device 193, and the pneumatic control port 405 are numbered by way of example.

[0088] Fig. 13shows a flow diagram of a method 1000 for operating a microfluidic device according to an embodiment. The device operated with the method can be the device described in the preceding figures. The method 1000 comprises a step 1005 of extracting sample material using the first sub-network. By way of example only, a sample substance is purified in the sub-network, and sample material such as DNA or RNA is extracted from the sample substance and transferred into the microfluidic network. This is optionally followed by a step 1010 of transferring sample material from the first sub-network to the second sub-network, wherein a solution with previously extracted sample material is transferred from the first sub-network to the second sub-network.In the optionally following step 1015 of amplifying sample material using the second subnetwork, sample material is amplified in the second subnetwork and thus made accessible for, for example, fluorometric detection. In a further embodiment of the method 1000, the extraction step 1005 or the transfer and amplification steps 1010, 1015 can be omitted. The first extraction step 1005 can be omitted, for example, if the species to be detected are already present in extracted form in an input sample fluid.The second step 1010 of transferring and the third step 1015 of amplifying can be omitted, for example, if only a purification or extraction of sample material is carried out in the device and the sample material is removed from the microfluidic device again, for example after the first step 1005 of extracting has been carried out.

[0089] Fig. 14 shows a flowchart of a method 1000 for operating a microfluidic device according to an embodiment. The method 1000 shown here corresponds to or is similar to the method described in the previous Figure 13described method, with the difference that the individual steps 1005, 1010, 1015 have additional sub-steps. In this exemplary embodiment, the extraction step 1005 comprises a mixing sub-step 1105 and a dissolving sub-step 1110 and a lysing sub-step 1115 and a filtering sub-step 1120 and a rinsing sub-step 1125. In this exemplary embodiment, the transferring step 1010 comprises a eluting sub-step 1130, and the amplifying step 1015 comprises a dissolving sub-step 1135 and a multiplying sub-step 1140 and a detecting sub-step 1145. In the mixing sub-step 1105, a liquid sample introduced into the sample introduction chamber is mixed with a binding buffer merely by way of example.The mixing is carried out in particular using the first functional module, the additional second functional module, the pump module, and a plurality of the following elements: microfluidic valves, microfluidic pump chambers, a sample input chamber, and a venting device. In this exemplary embodiment, the binding buffer is stored upstream in exactly one of the three pre-storage chambers of the first subnetwork. In this exemplary embodiment, the adjacent functional modules comprise microfluidic valves, which are closed in this sub-step 1105, thus preventing unwanted microfluidic crosstalk with the adjacent functional modules.

[0090] In sub-step 1110 of detachment, in this exemplary embodiment, sample material is detached within the sample input chamber, which sample material is initially connected to a sampling device inserted into the sample input chamber. The detachment takes place, by way of example only, by means of a transfer fluid that is introduced into the chamber, in particular using the first and second functional modules and the pump module and a plurality of the following elements: microfluidic valves, microfluidic pump chambers, pre-storage chambers, a sample input chamber, and a venting device. The transfer fluid is stored, by way of example only, in exactly one of the pre-storage chambers. The adjacent functional modules comprise, in particular, microfluidic valves, which are closed in this sub-step 1110, thus preventing undesired microfluidic crosstalk with the adjacent functional modules.

[0091] In sub-step 1115 of lysing, in this exemplary embodiment, lysis buffer is added to a sample substance introduced into at least one of the sample input chambers, and the sample substance or components thereof are tempered to effect thermally induced lysis of components of the sample substance. The components of the sample substance to be lysed in this sub-step 1115 are, by way of example only, bacteria. In another exemplary embodiment, they may be, for example, other cells. The lysing is carried out in particular using the first, second, and additional second functional modules and the pump module, as well as a plurality of the following elements: microfluidic valves, microfluidic pump chambers, pre-storage chambers, a sample input chamber, and a venting device. The lysis buffer is pre-stored in exactly one of the pre-storage chambers, by way of example only.The adjacent functional modules comprise, in particular, microfluidic valves, which are closed in this sub-step 1115, thus preventing unwanted microfluidic crosstalk with the adjacent functional modules. In an alternative embodiment of the lysis sub-step 1115, thermally induced lysis of components of the sample substance can be carried out by tempering the filter element after the components of the sample substance have been concentrated on the filter element in the filtering sub-step 1120.

[0092] In this exemplary embodiment, in sub-step 1120 of filtering, components of the sample substance introduced into at least one of the sample input chambers are filtered and bound using the filter element, so that the components of the sample substance are concentrated on the filter element. The filtering is carried out in particular using the third, fourth, second, or additional second functional module and the pump module(s), as well as a plurality of the following elements: microfluidic valves, microfluidic pump chambers, a sample input chamber, the filter element, the liquid storage chamber, and venting devices. The adjacent functional modules comprise, in particular, microfluidic valves, which are closed in this sub-step 1120, so that undesired microfluidic crosstalk with the adjacent functional modules is prevented in this sub-step.For example, a binding buffer with sample material is pumped from one of the sample input chambers via the filter element into the liquid storage chamber by means of the pump module, whereby components of the sample material, such as DNA or RNA, are bound to the filter element and concentrated. In this exemplary embodiment, the filtering sub-step 1120 takes place after the mixing sub-step 1105, the dissolving sub-step 1110, and the lysing sub-step 1115 have been carried out. In another exemplary embodiment, the filtering sub-step 1120 can take place before or after the lysing sub-step, depending on the precise embodiment of the method: If the lysis takes place in one of the sample input chambers, the filtering sub-step can be carried out after the lysing sub-step. If, however, the lysis takes place on the filter element, the filtering sub-step can be carried out before the lysing sub-step 1115.

[0093] In sub-step 1125 of rinsing, in this exemplary embodiment, the microfluidic network is rinsed with a wash buffer, for example, to remove residues of the binding buffer present in the microfluidic network after sub-step 1120 of filtering. Rinsing is carried out in particular using the first, third, and fourth functional modules and the pump module, as well as a plurality of the following elements: microfluidic valves, microfluidic pump chambers, pre-storage chambers, the liquid storage chamber, and a venting device. For example only, the wash buffer is pre-stored in exactly one of the pre-storage chambers.

[0094] In sub-step 1130 of elution, in this exemplary embodiment, sample material present on the filter element is dissolved in order to make it accessible for molecular diagnostic detection by means of an amplification reaction. For example only, the sample material is DNA. In another example, it can also be RNA, for example. For this purpose, for example only, an elution buffer is pumped over the filter element in order to transfer sample material present on the filter element into the elution buffer. In connection with the elution, in this exemplary embodiment, the elution buffer with sample material is also transferred from the first sub-network to the second sub-network via the microfluidic connecting channel.The elution is carried out in particular using the first, third, and fourth functional modules of the first subnetwork and the first functional module of the second subnetwork, as well as the pump module, as well as a plurality of the following elements: microfluidic valves, microfluidic pump chambers, microfluidic connecting channel, pre-storage chambers, liquid storage chamber, and venting device. The elution buffer is stored upstream in exactly one of the pre-storage chambers, merely by way of example.

[0095] In sub-step 1135 of dissolving, in this embodiment, the elution buffer with the contained sample material is used to dissolve a freeze-dried or lyophilized reagent, which can also be referred to as a reaction bead, and thus provide a reaction mix for a subsequent amplification reaction. This is merely an example of a master mix for carrying out a polymerase chain reaction. Dissolving is carried out in particular using the first and second functional modules of the second subnetwork as well as a plurality of the following elements: microfluidic valves, microfluidic pump chambers, microfluidic chamber, and venting device. The freeze-dried or lyophilized reagent or the reaction bead is present in a pump chamber or the passive microfluidic chamber in this embodiment.In this embodiment, the elution buffer containing sample material is pumped into at least one of the elements for dissolving the reaction bead using pump chambers. After dissolving the reaction bead, the resulting reaction mix is ​​transferred, for example only, from the second functional module of the second subnetwork to the first functional module of the second subnetwork.

[0096] In this exemplary embodiment, in sub-step 1140 of amplification, at least one amplification reaction is carried out to amplify sample material present in the reaction mix. The amplification is carried out in particular using the first functional module of the second sub-network and a plurality of the following elements: microfluidic valves, microfluidic pump chambers. The amplification is carried out, for example, by a polymerase chain reaction. In another exemplary embodiment, the amplification can also be carried out by an isothermal amplification method. As part of the sub-step of amplification, the microfluidic pump chambers in which the reaction mix is ​​present can thus also be tempered in order to create suitable physical conditions that enable the amplification reaction to proceed.

[0097] In this exemplary embodiment, in sub-step 1145 of detection, a signal is detected in order to detect the occurrence of at least one amplification reaction during sub-step 1140 of replication. The signal is, merely by way of example, an optical signal, for example a fluorescence signal emanating from at least one fluorescent dye, which indicates the occurrence of at least one amplification reaction. In another exemplary embodiment, sub-step 1145 of detection can occur simultaneously with sub-step 1140 of replication or after sub-step 1140 of replication. Optionally, in sub-step 1145 of detection, the signal or a variable derived therefrom can also be output in order to output the result of the sample analysis performed in the microfluidic analysis device to a user.

[0098] In further embodiments of the method, individual sub-steps can be omitted or repeated or swapped in order with other sub-steps, as described by way of example in the following embodiments.

[0099] Fig. 15 shows a flowchart of a method 1000 for operating a microfluidic device according to an embodiment. The method 1000 shown here corresponds to or is similar to the method described in the preceding Figures 13 and 14 described method. In this variant of method 1000, the substeps 1105 of mixing, 1115 of lysing, 1120 of filtering, 1125 of rinsing, 1130 of elution, 1135 of dissolving, 1140 of amplification, and 1145 of detection are carried out successively.

[0100] Fig. 16shows a flowchart of a method 1000 for operating a microfluidic device according to an embodiment. The method 1000 shown here corresponds to or is similar to the method described in the preceding Figures 13, 14 and 15 described method. In this variant of method 1000, the substeps 1110 of dissolving, 1120 of filtering, 1115 of lysing, 1125 of rinsing, 1130 of elution, 1135 of dissolving, 1140 of amplification, and 1145 of detection are carried out successively.

[0101] Fig. 17 shows a schematic representation of an embodiment of an analysis device 1400 for receiving a microfluidic device. The analysis device 1400 is designed merely as an example to receive a microfluidic device, as described in the preceding Figures 1 to 12described in order to carry out analysis processes within the device. In this embodiment, the analysis device 1400 comprises a control device 1410, which is designed to carry out the steps of the Figures 13 to 16 described method with respect to the device.

Claims

1. A microfluidic device (100) for analyzing sample material, wherein the device (100) comprises a microfluidic network (105) with a first sub-network (110) for purifying sample material, wherein the first sub-network (110) comprises a first functional module (250) for providing liquid reagents, a second functional module (261) for introducing a sample substance, a third functional module (270) for filtering sample material, a fourth functional module (280) for storing a liquid, and a first microfluidic pumping chamber (151) and a second microfluidic pumping chamber (152) for establishing fluid transport, characterized in that the first microfluidic pumping chamber (151) and the second microfluidic pumping chamber (152) are connected in series to the third functional module (270) for filtering sample material.

2. Microfluidic device (100) according to claim 1, with a second sub-network (120) connected to the first sub-network (110) by a connecting channel (125) for amplifying sample material, wherein the first sub-network (110) and the second sub-network (120) are separable from one another, wherein the second sub-network (120) comprises at least a first functional module (290) for amplifying sample material and a second functional module (295) for releasing at least one amplification reaction bead.

3. Microfluidic device (100) according to one of the preceding claims, wherein the first subnetwork (110) and the second subnetwork (120) are arranged in a linear topology.

4. Microfluidic device (100) according to one of the preceding claims, wherein the device (100) has a control port (718) for applying a first pressure to the first microfluidic pumping chamber (151) and a control port (709) for applying a second pressure to the second microfluidic pumping chamber (152) in order to effect a pressure difference at the third functional module (270) for filtering sample material by squeezing the first microfluidic pumping chamber (151) and sucking in the second microfluidic pumping chamber (152).

5. Microfluidic device (100) according to one of the preceding claims, wherein a first port of the first microfluidic pumping chamber (151) is connected to the first functional module (250) for providing liquid reagents and to the second functional module (261) for introducing a sample substance, a second port of the first microfluidic pumping chamber (151) is connected to a first port of the third functional module (270) for filtering sample material, a first port of the second microfluidic pumping chamber (152) is connected to the second functional module (261) for introducing a sample substance, and a second port of the second microfluidic pumping chamber (152) is connected to a second port of the third functional module (270) for filtering sample material.

6. The microfluidic device (100) according to claim 5, wherein the third functional module (270) comprises a filter element (170) detachably connected to the first port of the third functional module (270) and detachably connected to the second port of the third functional module (270), and at least one microfluidic valve (139, 140') connected between the first and second ports of the third functional module (270).

7. Microfluidic device (100) according to one of claims 5 to 6, wherein the second functional module (261) comprises a sample input chamber (167) for inputting a sample substance, wherein the sample input chamber (167) is detachably connected to a first port of the second functional module (261) and detachably connected to a second port of the second functional module (261), wherein the first port of the first microfluidic pumping chamber (151) is connected to the first port of the second functional module (261) and the first port of the second microfluidic pumping chamber (152) is connected to the second port of the second functional module (261).

8. Microfluidic device (100) according to claim 2, wherein the device (100) has a common control port (703) for applying a pressure, wherein the common control port (703) is connected by a first control channel (431) to an element of a functional module (250, 261, 270, 280, 580) of the first sub-network (110) and by a second control channel (432) to an element of a functional module (290, 295) of the second sub-network (120).

9. Microfluidic device (100) according to claim 2, wherein the first sub-network (110) has a first shut-off valve (144) for closing the connecting channel (125) and / or the second sub-network (120) has a second shut-off valve (145) for closing the connecting channel (125).

10. Microfluidic device (100) according to claim 9, comprising a valve control port (713) for applying a pressure, wherein the first shut-off valve (144) and the second shut-off valve (145) are controllable by means of the valve control port (713).

11. Microfluidic device (100) according to one of the preceding claims, wherein the fourth functional module (280) of the first sub-network (110) is designed to dispense a liquid with sample material.

12. Microfluidic device (100) according to one of the preceding claims, wherein the first sub-network (110) has an additional fourth functional module (580) for storing a liquid, wherein the first microfluidic pumping chamber (151) is connected in parallel to the additional fourth functional module (580).

13. A method (1000) for operating a microfluidic device (100) according to any one of the preceding claims, wherein the method (1000) comprises the following steps (1005, 1010, 1015): extracting (1005) sample material using the first subnetwork (110); characterized in that the step (1005) of extracting comprises a sub-step (1120) of filtering, in which a pressure difference is generated at the third functional module (270) by combined squeezing of the first microfluidic pumping chamber (151) and suction with the second microfluidic pumping chamber (152) in order to effect the filtering of sample material.

14. The method (1000) according to claim 13, comprising a step of transferring (1010) sample material from the first sub-network (110) to the second sub-network (120) and a step of amplifying (1015) sample material using the second sub-network (120).

15. The method according to claim 14, wherein the step (1005) of extracting comprises a sub-step (1105) of mixing and / or a sub-step (1110) of dissolving and / or a sub-step (1115) of lysing and / or a sub-step (1125) of rinsing and / or wherein the step (1010) of transferring comprises a sub-step (1130) of eluted and / or wherein the step (1015) of amplifying comprises a sub-step (1135) of dissolving and / or a sub-step (1140) of amplifying and / or a sub-step (1145) of detecting.

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