WO2018192809A1

WO2018192809A1 - Apparatus and method for a microfluidic system for analyzing a sample

Info

Publication number: WO2018192809A1
Application number: PCT/EP2018/059217
Authority: WO
Inventors: Julian Kassel; Daniel Czurratis
Original assignee: Robert Bosch Gmbh
Priority date: 2017-04-18
Filing date: 2018-04-11
Publication date: 2018-10-25
Also published as: DE102017206489A1

Abstract

The invention relates to a lab-on-chip cartridge (100) for a microfluidic system for analyzing a sample, wherein the lab-on-chip cartridge (100) has a reagent receptacle (104) having a multi-layer structure, wherein an integrated cavity (108) and a flexible membrane (110) are arranged in the multi-layer structure, wherein the flexible membrane (100) separates at least two substrate layers (106) from one another. Furthermore, the lab-on-chip cartridge (100) comprises an insert part (114) which is arranged in the integrated cavity (108), wherein the insert part (114) contains a liquid reagent (116) and is sealed with a barrier film (118). Lastly, the lab-on-chip cartridge (100) comprises a plunger (102) for releasing the liquid reagent (116) from the insert part (114), wherein the plunger (102) is arranged opposite the reagent receptacle (104), wherein an end face of the plunger (102) has at least one planar portion for pushing in the flexible membrane (110).

Description

Description title

Apparatus and method for a microfluidic system for analyzing a sample

State of the art

The invention is based on a device or a method according to the preamble of the independent claims. The subject of the present invention is also a computer program.

On chip laboratory cartridges, the whole functionality of a

housed on a credit card-sized plastic substrate, whereby complex biological, diagnostic, chemical or physical processes can be miniaturized.

Disclosure of the invention

Against this background, with the approach presented here, a method, furthermore a control unit, which uses this method, and finally a corresponding computer program according to the main claims are presented. The measures listed in the dependent claims are advantageous developments and improvements in the independent

Claim specified device possible.

A liquid reagent for a chip laboratory cartridge can be stored for long-term stability and reproducibly provided if necessary by the method of a mechanical element, such as a plunger, this process being independent of the orientation or inclination of the chip laboratory cartridge. It will be a chip laboratory cartridge for a microfluidic system for

Analyzing a sample (for example, a diagnostic material), the chip cartridge having the following features: a reagent receptacle having a multilayer structure, wherein an integrated cavity and a flexible membrane are arranged in the multi-layer structure, the flexible membrane at least two (polymer) ) Separates substrate layers from each other; an insert disposed in the integrated cavity, wherein the

Insert contains a liquid reagent and is sealed with a barrier film; and a plunger for releasing the liquid reagent from the insert, wherein the plunger is disposed opposite to the reagent receptacle, wherein an end face of the plunger has at least one planar portion for pressing the flexible membrane.

A chip laboratory cartridge may be a microfluidic device in which a liquid reagent is provided or transported on a chip. In particular, the chip laboratory cartridge may be suitable for so-called lab-on-chip products and medical devices which receive samples of diagnostic or diagnostic material and

then process automatically. The diagnostic material may be analytical material for complex biological, chemical or physical processes.

Many chip lab cartridges require a variety of liquid reagents, such as saline solutions, ethanol-containing solutions, aqueous solutions

Solutions, detergents or dry reagents, such as lyphilisates or salts, which are required for a wide variety of diagnostic applications. These

Reagents can be manually pipetted onto the chip laboratory cartridge or pre-stored on the cartridge. The latter brings advantages in terms of automation, contamination risks, user friendliness and transportability of chip laboratory cartridges.

The chip laboratory cartridge can furthermore be a chip which has a multilayer construction, that is to say it is composed of at least two layers, which are made of polymer-based substrates. The (polymer)

Substrate layers may be separated by a flexible membrane. The flexible membrane is adapted to be deflected upon pressure on the same. The flexible membrane is designed tear-resistant, d. H. it is so stable that it does not tear when pressed by the plunger.

The multi-layer structure of the chip may include cavities in the form of chambers and channels. The (polymer) substrate layer can be made of plastics with high barrier properties, such as COP, COC, PP, PE or PET. Concepts based on such plastics can be integrated directly into the material system of the chip laboratory cartridge or can be fluidly connected to the chip laboratory cartridge by means of a joining process such as gluing, welding or clamping. The insert may be an injection molded chamber within a sealed cavity in the (polymeric) substrate layer, the insert containing a liquid reagent for analysis of the material to be tested. The insert can be glued, clamped, welded or integrated by other joining methods in the chip laboratory cartridge, which eliminates process steps in the production and / or simplified.

The insert may be sealed with a barrier film. By means of such a barrier film, the liquid reagent can be stored safely in the fluid chamber and can be selectively released only when required by introducing the stamp unit or the plunger into the barrier film.

Furthermore, the insert may be formed so that it can be accommodated accurately in the reagent receptacle, wherein the material of the insert may have a higher barrier property with respect to the liquid reagent, as the (polymer) substrate layer. Thus, different fluids with different requirements for long-term stable storage in special for the requirements of the fluids molded inserts safely stored in the chip laboratory cartridge.

The plunger is advantageous with at least one planar section for

Pressing the flexible membrane equipped to ensure the most efficient squeezing out of the liquid reagent from the insert, the insert may now be configured angular to an existing

Make optimum use of the volume of construction in the reagent receptacle.

According to one embodiment, the (polymer) substrate layer of the

Reagent receptacle have at least one breakthrough. The breakthrough serves as a window to give the plunger an entry way into the

Open reagent receptacle to release the liquid reagent, the ram passes through the opening for reagent release with the plunger.

According to one embodiment, the plunger may be cylindrically shaped. By a simple cylindrically designed shape of the plunger forces required to break the barrier film are reduced, which

Cost savings in the design of the overall system allows. In its simplest embodiment as a cylindrically shaped form such a plunger can also be produced very inexpensively.

According to one embodiment, the end face of the plunger a

have circular segment-shaped recess. The advantage of this

circular segment-shaped recess in the plunger is in particular that after the tearing of the barrier film, the flexible membrane according to the design of the ram sets and thus the fluidic path can not be blocked. At the same time it is ensured by the advantageous circular segment-shaped recess of the plunger that just in this area the torn barrier film can not block the fluidic path as well, so that the liquid reagent can be efficiently expressed from the insert. According to one embodiment, the end face of the plunger a

(for example in the direction of the flexible membrane) obliquely

Have surface. The obliquely extending end face of the plunger then has, for example, an angle between 0.1 ° and 30 ° with respect to a

Movement axis of the plunger in the direction of the Einlegteils. The use of a plunger with a sloping surface leads to a

Stress exaggeration of the barrier film at the front edge of the plunger and thus ensures an initial crack propagation.

According to one embodiment, the plunger may have a projection adjacent to the end face. The plunger with an adjacent projection on the front side is advantageously suitable for pressing out the liquid reagent in the edge regions of the insert as well as for a multi-stage process for pressing out the liquid reagent. So can the ram with projection

for example, be retracted first to tearing the barrier film. After retracting the plunger it is then rotated, for example, whereby the projection can not block the fluidic path and the

Liquid reagent is emptied efficiently.

According to one embodiment, the flexible membrane can tear during

Pressing the plunger be formed. The flexible membrane allows a complete fluidic separation between the liquid reagent and the

mechanical pestle. This has the particular advantage that the

Material selection of the chip laboratory cartridge independent of the

Requirements of long-term stable reagent pre-storage remains. The flexible membrane also allows a reproducible displacement of the liquid in the adjacent cavity with high transfer efficiency.

There is further provided a method of releasing a liquid reagent using the chip laboratory cartridge, the method comprising the steps of:

Providing the liquid reagent, wherein the plunger is pressed onto the flexible membrane and penetrates to a predetermined depth to tear the barrier film; and Opening a fluidic path, wherein the liquid reagent is displaced into the cavity of the (polymer) substrate layer to release the liquid reagent.

According to one embodiment, in the method for releasing a

Liquid reagent of the plunger from the reagent receptacle initially retracted and then pressed again into the reagent receptacle. Through this multi-stage method of moving the mechanical plunger, the liquid reagent to be released receives a fluidic preferred direction, so that the reproducibility and the efficiency of the

Reagent release can be significantly increased. The manipulation of the fluidic preferred direction also allows a reagent release at angles from 0 ° to 90 °. This allows for enormous degrees of freedom in the design and processing of the entire chip laboratory cartridge.

According to one embodiment, in the method for releasing a

Liquid reagent of the plunger are rotated along its axis of movement. By a combination of translational and rotating process paths of the mechanical plunger, the emptying efficiency can be further increased, in particular for a highly viscous liquid reagent.

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.

The approach presented here also provides a control unit which is designed to implement the steps of a variant of a method presented here

to implement, control or implement appropriate facilities. Also by this embodiment of the invention in the form of a control device, the object underlying the invention can be achieved quickly and efficiently.

For this purpose, the control unit can have at least one arithmetic 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 arithmetic unit may be, for example, a signal processor, a microcontroller or the like, wherein the memory unit may be a flash memory, an EEPROM or a magnetic memory unit. The communication interface can be designed to read or output data wirelessly and / or by line, wherein a communication interface that can read or output line-bound data, for example, electrically or optically read this data from a corresponding data transmission line or output in a corresponding data transmission line.

In the present case, a control device can be understood as meaning an electrical device which processes sensor signals and outputs control and / or data signals in dependence thereon. The control unit may have an interface, which may be formed in hardware and / or software. In a hardware training, the interfaces may for example be part of a so-called system ASICs, the various functions of the

Control unit includes. However, it is also possible that the interfaces are their own integrated circuits or at least partially consist of discrete components. In a software training, the interfaces may be software modules that are present, for example, on a microcontroller in addition to other software modules.

Also of advantage is a computer program product or computer program with program code which 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 for the implementation, implementation and / or

Triggering the steps of the method according to one of the above

described embodiments, in particular when the program product or program is executed on a computer or a device. Embodiments of the approach presented here are shown in the drawings and explained in more detail in the following description. It shows:

1 shows a schematic representation of a chip laboratory cartridge with a plunger according to an embodiment;

Fig. 2 is a schematic representation of a cylindrical plunger penetrating into a reagent receptacle, according to one embodiment;

Fig. 3 is a schematic representation of a plunger with an adjacent projection on its end face, which penetrates into a Reagenzaufnahmebehälter, according to an embodiment;

Fig. 4 is a schematic representation of a plunger having a tapered surface penetrating into a reagent receptacle, according to one embodiment;

Fig. 5 is a schematic representation of a plunger with a

circular segment-shaped recess in the front side, in a

Reagent receptacles penetrate according to one embodiment;

Fig. 6 is a schematic representation of a plunger, in a

Reagent receptacle penetrates, according to an embodiment;

7 shows a schematic sequence of a cylindrically shaped plunger which penetrates into a chip laboratory cartridge according to an embodiment; and

8 is a flowchart of a method for releasing a

Liquid reagent using the chip laboratory cartridge according to an embodiment.

1 shows a schematic representation of a chip laboratory cartridge 100 with an opposing tappet 102 according to one exemplary embodiment. In the left part of the illustration, the plunger 102 and its orientation relative to a Reagenzaufnahmebehälters 104 are shown in a multi-layer structure at an angle of 90 °. In the right part of the illustration, the plunger 102 and its orientation with respect to the reagent receptacle 104 are shown at an angle of 30 °.

In order to increase in particular the reproducibility and efficiency of the emptying in the reagent release, on the one hand, the geometry of the plunger 102 used and on the other hand the procedure, such as a method direction, a process speed and a rotation of the

Pestle 102, of importance. In addition, it may be in terms of

Of interest to the end user that the reagent release in the multi-layer structure should be performed at different angles from 0 ° to 90 °. In particular, at angles smaller than 90 °, the decreasing part of the gravity-driven evacuation may be achieved by an advantageous

Design of the plunger 102 and its process movement,

In particular translation and / or rotation, be compensated so that a reliable reagent release is made possible. FIG. 2 shows a schematic representation of a cylindrical plunger 102 which penetrates into a reagent receptacle 104 of a chip laboratory cartridge 100 according to an exemplary embodiment. The illustration shows four individual

Illustrations of which the left figure shows a plan view of the plunger 102 and the remaining three images each show a transverse view in a sequence of the plunger 102, as this in the reagent receptacle 104 of

Chiplabor- cartridge 100 retracts.

The chip laboratory cartridge 100 comprises a reagent receptacle 104, which in turn is composed of a multi-layer structure. The multilayer structure consists of at least two (polymer) substrate layers

106. In at least one of the (polymer) substrate layers 106 of the

Multilayer construction is additionally integrated with at least one cavity 108.

At least one of the (polymer) substrate layers 106 is connected to a flexible membrane 110 and another (polymer) substrate layer 106. At least one (polymer) substrate layer 106 has at least one Breakthrough 112 in which the flexible membrane 110 is not connected to the (polymer) substrate layer 106 and therefore is movable. The aperture 112 is formed through an opening of one of the substrate layers 106. In at least one (polymer) substrate layer 106 are located in a recess an additional insert 114, which ideally has higher barrier properties compared to the liquid reagent than the surrounding (polymer)

Substrate layer 106 and thus a long-term stable storage of a

Liquid reagent 116 is guaranteed. The insert 114 contains the liquid reagent 116 and is provided with a barrier film

118 sealed. As can be seen in the middle image of the illustration, for the release of the liquid reagent 116, a plunger 102 is pressed in through the opening 112, deflecting the flexible membrane 110 without it tearing and breaking the barrier film 118. In this case, the liquid reagent 116 is displaced via the fluidic path 120 into the integrated cavity 108 and now stands

(Corresponding to the right image of the representation from FIG. 2) for further microfluidic processes which will not be described in detail here.

3 shows a schematic representation of a plunger 102 with an adjacent projection 304 on its end face 302, which in one of the

Reagent receptacle 102 of the chip laboratory cartridge 100 according to an embodiment penetrates. The illustration shows four individual images, of which the left-hand illustration shows a plan view of the tappet 102 and the remaining three images each show a transverse view in a sequence of the tappet 102 as it is inserted into the reagent receptacle 104 of the chip laboratory.

Cartridge 100 retracts.

At angles of the reagent container 102 to the horizontal, which are smaller than 90 °, the proportion of gravity decreases in the

Reagent release increasingly. Therefore, in an advantageous

Embodiment of the mechanical plunger 102 is provided with a projection 304 on the end face 302 in order to promote the tearing of the barrier film 118 in the region of the fluidic path 120. Thus, the highest stresses initially concentrate in this region, which leads to premature tearing of the barrier film 118 in this area and thus the fluidic path 120 in FIG the integrated cavity 108 releases. In this way, a certain preferred direction of the released liquid reagent 116 is generated, even in the case of a

On the one hand this increases the emptying efficiency of the liquid reagent 116 from the insert 114 and on the other hand it prevents any possible crosstalk of adjacent chambers in a chamber structure that are fluidly interconnected.

4 shows a schematic representation of a tappet 102 with an inclined surface 402, which penetrates into a reagent receptacle 102 of a chip laboratory cartridge 100 according to one exemplary embodiment. The illustration shows four individual images, of which the left-hand illustration shows a plan view of the plunger 102 and the remaining three images each show a transverse view in a sequence of the plunger 102, as shown in FIGS

Reagent receptacle 104 of the chip laboratory cartridge 100 retracts.

In a further advantageous embodiment of the plunger 102, the end face 302 of the plunger is provided, for example, at an angle of 0.1 ° to 30 ° with respect to a movement axis of the plunger 102. The obliquely extending on the flexible membrane 110 surface 402 leads to a

Voltage overshoot of the barrier film 118 at the end face 302 of the plunger 102 and provides an initial crack propagation in this area. This produces a certain preferred direction of the released liquid reagent 116 even with decreasing proportion of gravity due to inclination of the chip laboratory cartridge 100. Smaller angles of the end face 302 thus allow a higher evacuation efficiency of the chambers, since the shape of the plunger 102 better in the shape of the chambers can add. On the other hand, larger angles allow cracking of the barrier film 118 at lower forces, and accordingly, the draining efficiency may decrease. This emptying efficiency can be improved by adjusting the shape or slope of the end face 302 of the plunger 102 according to the design of the insert 114. In addition, these structures are used to break up the

Barrier film 118 required forces reduced, which allows cost savings in the design of the overall system. 5 shows a schematic representation of a plunger 102 with a circular segment-shaped recess 502 in the end face 302, which penetrates into the reagent receptacle 104 of a chip laboratory cartridge 100 according to one exemplary embodiment. The illustration shows four individual images, of which the left-hand illustration shows a plan view of the plunger 102 and the remaining three images each show a transverse view in a sequence of the plunger 102 as it enters the reagent receptacle 104 of the chip laboratory cartridge 100. In a further advantageous embodiment, the plunger 102 is designed so that in the end face 302 of the plunger 102 is a semicircular or

circular segment-shaped recess 502 is located. This recess 502 extends along a portion of a side wall of the plunger 102. In order to further increase the tension for a controlled crack propagation in this area, the end face 302 of the plunger 102 can additionally with a

Angle of 0.1 ° to 30 ° are provided. The advantage of the semicircular recess 502 of the end face 302 of the plunger 102 is, in particular, that after the tearing of the barrier film 118 on the formation of the end face 302 of the plunger 102 corresponding to the flexible membrane 110 sets around the plunger 102 thus the fluidic path 120 is not blocked can be.

At the same time, however, the advantageous semicircular or circular segment-shaped recess 502 of the end face 302 of the tappet 102 ensures that precisely in this region the torn barrier foil 118 likewise can not block the fluidic path 120 or the probability of a blockage is reduced.

Fig. 6 shows a schematic representation of a plunger, which in a

Reagent receptacle 104 a chip laboratory cartridge 100 according to an embodiment penetrates. The illustration shows four individual figures, which show a transverse view of a sequence of the plunger 102, as this in the

Reagent receptacle 104 of the chip laboratory cartridge 100 retracts.

In addition to the design of an advantageous embodiment of the plunger 102, the procedure of the mechanical plunger 102 plays a crucial role for a reproducible and efficient reagent release. Next to the previously single-stage retraction of the plunger 102 in the multi-layer structure of the reagent receptacle 104, can also be a two- or multi-stage

Retracting the plunger 102 to be beneficial. For example, after the tearing foil 118 tears, the tappet 102 is returned to its original position. On the one hand, by the retraction of the plunger 102, depending on the angle of the system, already a gravity-driven

Emptying of the liquid reagent 116 favors, since the flexible membrane 110, the fluidic path 120 in the direction of the integrated cavity 108 immediately after the deflection of the flexible membrane 110 completely free again. On the other hand, this backfilling has the advantage that the liquid reagent 116 experiences a certain suction direction as a result of the volume released, depending on the angle of the system, and efficient re-introduction of the tappet 102 enables efficient emptying with preferential direction of the liquid reagent 116. This is particularly advantageous for highly viscous liquid reagents. In this multi-stage process, the already presented advantageous

Formations of the end face 302 or the side edge of the plunger 102 are also used.

7 shows a schematic representation of a cylindrically shaped plunger 102, which penetrates into a reagent receptacle 104 of a chip laboratory cartridge 100 according to one exemplary embodiment. The illustration shows six individual images, of which the left-hand illustration shows a plan view of the plunger 102 and the remaining five images each show a transverse view in a sequence of the plunger 102, as shown in FIGS

Reagent receptacle 104 of the chip laboratory cartridge 100 retracts.

In addition to the different variants in the translational movement of the plunger 102, rotating movements and combinations of both forms of movement are also advantageous for reliable reagent release. In an advantageous embodiment, the plunger 102 with the projection 304 on its end face 302 first into the reagent receptacle 104th

retracted to the tearing of the barrier film 118 in the region of the fluidic path 120. After moving back the plunger 102, this is rotated by 180 ° about its axis of movement, for example. This allows the projection 304 the fluid path 120 and the liquid reagent 116 can be efficiently emptied.

By multiple translational or rotating movements of the plunger 102, the stored Flüssigreagenz 116 can also before the actual

Processing be well mixed again. This may be particularly advantageous with a liquid reagent 116, such as a soap-like solution, brine, or detergent, in which solids may precipitate or settle over time. Here, for example, the plunger 102 initially only partially into the integrated cavity 108 and on the flexible membrane

110 pushed retracting and then back and forth several times. If the plunger 102 is finally moved to the bottom of the cavity 108 or of the insert, the well-mixed liquid reagent 116 is completely released into the integrated cavity 108.

8 shows a flowchart of a method 800 for releasing a liquid reagent using the chip laboratory cartridge according to an exemplary embodiment. In a step of providing 801 of the upstream Flüssigreagenz the plunger is used, which can penetrate through at least one opening in the (polymer) substrate layer of the reagent container by a translational, rotating or a combination of both in the chip laboratory cartridge. Here, the flexible membrane is deflected by the plunger without tearing. Upon contact with the barrier film, a force is applied by the plunger, which leads to tearing of the barrier film. In a step 803, the fluidic path for the liquid reagent is now opened. By one or more stages of the method of the mechanical plunger and an advantageous construction of the plunger and / or the end face of the plunger is a

Generated preferred direction of the released liquid reagent and prevents blocking of the fluidic path through the flexible membrane.

If an exemplary embodiment comprises an "and / or" link between a first feature and a second feature, then this is to be read so that the embodiment according to an embodiment is both the first feature as well as the second feature and according to another embodiment, either only the first feature or only the second feature.

Claims

claims

1. Chip laboratory cartridge (100) for a microfluidic system for

Analyzing a sample, the chip lab cartridge (100) comprising: a reagent receptacle (104) having a multi-layer structure, wherein in the multi-layer structure an integrated cavity (108) and a flexible membrane (110) are arranged, the flexible membrane (10) 110) separates at least two substrate layers (106) of the multi-layer structure from each other; an insert (114) disposed in the integrated cavity (108) of one of the substrate layers (106), the insert (114) containing a liquid reagent (116) and sealed with a barrier film (118); and a plunger (102) for releasing the liquid reagent (116) from the insert (114), wherein the plunger (102) is disposed opposite to the reagent receptacle (104), wherein an end face (302) of the plunger (102) is at least one planar Part section for pressing the flexible membrane (110).

2. The chip laboratory cartridge (100) according to claim 1, wherein one of the

Substrate layers (106) of the Reagenzaufnahmebehälters (104) via at least one opening (112) for the passage of the plunger (102) has.

3. chip laboratory cartridge (100) according to one of the preceding

Claims, wherein the end face (302) of the plunger (102) has a circular segment-shaped recess (402). Chip laboratory cartridge (100) according to one of the preceding

Claims, wherein the end face (302) of the plunger (102) has a sloping surface (402).

Chip laboratory cartridge (100) according to one of the preceding

Claims, wherein the plunger (102) adjacent to the end face (302) has a projection (304).

Chip laboratory cartridge (100) according to one of the preceding

Claims, wherein the flexible membrane (110) is tear-resistant when depressing the plunger (102) is formed.

A method (800) for releasing a liquid reagent (116) using the chip lab cartridge (100) according to claims 1 to 6, said method (800) comprising the steps of:

Providing (801) the liquid reagent (116), wherein the plunger (102) is pressed onto the flexible membrane (110) and penetrates to a predetermined depth to tear the barrier film (118); and

Opening a fluidic path (120), wherein the liquid reagent (116) is displaced into the integrated cavity (108) of one of the substrate layers (106) to release the liquid reagent (116).

A method (800) for releasing a liquid reagent (116) according to claim 7, wherein the plunger (102) is initially retracted from the reagent receptacle (104) and then reentered into the reagent receptacle (104)

Reagent receptacle (104) is pressed.

A method (800) for releasing a liquid reagent (116) according to claim 7 or 8, wherein the plunger (102) is rotated about its axis of motion.

A controller adapted to execute and / or drive steps of the method (800) according to any one of the preceding claims in respective units. A computer program adapted to perform the method (800) of any one of the preceding claims.

A machine readable storage medium storing the computer program of claim 11.