Method for filling a microfluidic device using a dispensing system and corresponding test system

[0001] The invention relates to a method for filling nozzles of a dispensing system of a test system, and a test system, comprising a dispensing system and a microfluidic device and at least one photosensitive sensor with a large number of photodetectors.

[0002] Many point-of-care tests for in vitro diagnostics require the manual introduction of various solutions into a sample vessel. For example, WO 2012/080339 A1

1O shows a measuring device in which three different reagents must be instilled manually and sequentially in the course of the test in a volume range of 10 μ | This manual instillation is inconvenient because the manual work time during processing of the test is very long and binds the work force. In addition, manual instillation, due to the low strength and repeatability, creates some risk of producing erroneous results. In addition, the reagent dosing device is also described in WO 2012/080339 A1, in the form of a blister pack.

[0003] Automatic dosing devices solve the problems of manual reagent addition. They enable the quick and accurate introduction of various volumes into the sample vessel. Systems for in vitro diagnostics are often used to enable non-contact and very precise volume dispensing in the micro- to nanolitre range.

[0004] Descriptions US 2010/0221704 A1, US 2003/0132112 A1 and US 6,589,790 B1 each disclose a method for introducing solutions into microfluidic devices. Traditional test systems for many reagents, however, use expensive and complex components that are not suitable for on-site testing systems.

[0005] In traditional automatic dispensing systems with only one nozzle, with an outlet, and multiple valves (also multi-channel) for the flow of reagent needed, dead volumes remain on the fluid path where the reagents mix. Here, after each reagent, complete rinsing must occur and the flushing liquid must be collected in a separate container. However, it is inconvenient to place the proprietary nozzle flushing circuit in the care system at the care site. [0006] If, moreover, the liquids to be dispensed cannot contact each other prior to dispensing, because otherwise biochemical reactions may occur in the fluid system instead of the measuring chamber, separate fluid paths must be placed. For such a system, it is particularly important to ensure that prior to dispensing, there is no air in the fluid path that could affect the amount of reagent dispensed.

[0007] Therefore, it is the object of the present invention to ensure that the fluid path, before starting the actual measuring process, is completely filled with the reagent up to the outlet opening.

[0008] The object of the invention is solved in each case alone by a method according to claim 1, for filling a microfluidic device, in particular nozzles of a dispensing system of a test system, comprising a dispensing system and a microfluidic device, and a test system according to claim 9, comprising at least one dispensing system, microfluidic device and at least one photosensitive sensor with a large number of photodetectors.

[0009] In this connection, it has proved to be advantageous that it can be ensured that the solution is in the nozzle, and thus the correct solution volume can be supplied from the nozzle for the next process steps, without having to refill the nozzle, rinse , washing, and so on.

[001 O] According to the development, the dosing system contains at least two containers with solutions, with the first solution in the first container, especially chemiluminescent solution, and in the second container another solution, preferably an enzyme solution, and the containers via a fluid line are connected to at least one micropump, and immediately after that at least two nozzles are placed, the first nozzle is for applying the first solution, especially chemiluminescent solution, and the second nozzle for applying another solution, preferably an enzyme solution, into the hole for applying samples of the microfluidic device, comprising the following stages:

- transporting the first solution, especially the chemiluminescent solution, and the further solution, especially the enzyme solution, from each container through each nozzle of the dosing system to the opening for applying the sample of the microfluidic device, by means of at least one micropump,

- measuring the light signal, especially the chemiluminescent signal, by the reaction of the first solution, especially the chemiluminescent solution, and the further solution, especially the enzyme solution, in the measuring area of ​​the microfluidic device, by means of at least one photosensitive sensor with a large number of photodetectors,

- switching off at least one micropump when a light signal is detected, especially a chemiluminescent signal.

[0011] If a light signal can be detected in the microfluidic channel, and thus the first solution, especially the chemiluminescent solution, and the second solution, preferably the enzyme solution, react with each other because the signal is formed only by the reaction of both reagents. Because the contact and thus the reaction of both solutions is only possible after delivery from each nozzle, so both the first solution, especially the chemiluminescent solution, as well as the further solution, preferably the enzyme solution, must pass through the nozzles. By detecting the light signal that is formed from the chemical or biochemical reaction of both reagents, it is ensured that both the first solution, especially the chemiluminescent solution, as well as the further solution, preferably the enzyme solution, has reached the measuring area of ​​the microfluidic channel and is therefore also in dispensing nozzles that transport liquids to the microfluidic device through the sample application opening. At the same time, it turns out to be advantageous that it can be ensured that both the nozzle for applying the first solution, especially the chemiluminescent solution, as well as the nozzle for the further solution, preferably the enzyme solution, is filled with each liquid up to the outlet end, and thus during dispensing the correct volume is delivered, which is set at the time of dispensing. For carrying out various analyzes using a test system, it is important that prior to measurement it is ensured that the liquid meniscus in the dispensing device is at the end of the injection nozzles. If the meniscus of the first solution, especially the chemiluminescent solution, or the further solution, preferably the enzyme solution, was located further behind in the nozzle's liquid channel, then there would be no adequate amount

Of the solution during the measurement process and erroneous results occur during analysis by means of the test system due to insufficiently dosed volumes of liquid. It is also advantageous that thanks to this a cost-effective and high reproducibility system is also available. In the method according to the invention, in the test system according to the invention, the drawback of the prior art of using an additional container for a liquid path flushing solution is eliminated. To determine the filling of the nozzles up to the end of the outlet, no additional sensors are needed to prevent the compact design of the on-site testing system.

[0012] According to the development, it is provided that

O (a) first solution, especially chemiluminescent solution, is transported through the first nozzle to the microfluidic device sample application opening by means of at least one micropump, and then sucked into the measuring area of ​​the microfluidic device, (b) the measurement of the light signal takes place in the measuring area of ​​the microfluidic device by means of a world / eye / ego sensor with a large number of photodetectors, (c) the micropump switches off when the light signal changes, especially when the optofluid lens effect increases, (d) another solution, especially the enzyme solution, is transported through a further nozzle to the hole for applying a sample of the microfluidic device using a micropump, and then sucked into the measuring area of ​​the microfluidic device, (e) light signal, especially a chemiluminescent signal, is detected in the measuring area of ​​the microfluidic device using a photosensitive sensor with a large number of photodetectors, (f) the micropump switches off when a light signal is detected, especially the chemiluminescent signal.

[0013] The sequential application of solutions has the advantage that the presence of a first solution, especially a chemiluminescent solution, can be controlled, regardless of the subsequent solution, preferably the enzyme solution. In this way, in the absence of a light signal, especially the chemiluminescent signal, the source of the error can be found quickly, through the nozzle no sample was delivered to the sample application opening, relatively few first solution, especially chemiluminescent solution, either none or too / for another solution, preferably an enzyme solution, [0014] When the first solution is transported to the microfluidic channel, the light signal changes due to the optofluid lens effect, especially increases thanks to which the meniscus of the first solution can be measured.

[0015] In accordance with the development, further steps are carried out, wherein (g) the additional solution, especially the washing solution, is transported through another nozzle to the sample application opening of the microfluidic device by means of a micropump, and then sucked into the measuring area of ​​the microfluidic device,

(H) the offset of the previously measured light signal is measured in the measuring area of ​​the microfluidic device by means of a photosensitive sensor with a large number of photodetectors, (i) the micropump switches off when a light signal shift is detected.

[0016] This results in the fact that in the test system it can also be ensured that the nozzle is filled with an additional solution, in particular a washing solution, before the additional solution is dispensed during the actual analysis.

[0017] Contact of the solutions, especially the enzyme solution, with air may lead to clogging of the nozzle outlet. However, the outlet openings of the nozzles may also dry out as a result of them receding in them

An incorrect sample volume is provided about the liquid meniscus and in the subsequent analysis. Therefore, preferably, the outlet openings of the nozzles, after switching off the micropump, are closed by at least one sealing device, especially hermetically.

[0018] The combination of the micropump with the nozzle has the advantage that a stream is produced from nanolitre drops of each liquid, whereby targeted and contactless delivery of the respective solution from the nozzle outlet to the sample application opening of the microfluidic device is enabled. Targeted delivery is necessary to ensure that the entire volume of each solution is actually applied to the sample application opening of the microfluidic device to obtain the correct analysis result. Thanks to the micropump, the dosing of a few microliters into the application opening of the sample of the microfluidic device is obtained with an accuracy of mit + 1 μ |

[0019] After filling the nozzle of the dispensing system, the biological sample is introduced into the sample application opening and is transported to the microfluidic channel, where the target molecules of the biological sample react with the molecules through specific binding sites, which are located in the measuring sections of the measuring area, and by adding solutions from the dosing system, a chemical reaction takes place, relatively biochemical under light emission conditions, and a light signal is created, which is detected by a photosensitive sensor with a large number of photodetectors, so that a rapid assessment of the biological sample can occur, because many samples can be measured in succession, without intermediate rinsing.

[0020] In addition, according to the development, that the dispensing system may contain at least two containers with solutions, with the first container containing the first solution, especially chemiluminescent solution, and the second container contains another solution, preferably an enzyme solution, and the containers via a fluid line are connected to at least one micropump, and immediately after that at least two nozzles are placed, the first nozzle is for applying the first solution, especially chemiluminescent solution, and the second nozzle for applying another solution, preferably an enzyme solution, into the opening for applying the sample of the microfluidic device, thanks to which it is possible to quickly pour systems for testing.

[0021] According to the development, it is provided that the containers have a connector with a port for connection to a standard Luer cone of the fluid conduit to create a separable connection through the liquid conduit with the micropump. Thanks to this, bubble-free container docking can be obtained. In addition, it is preferred that the empty containers, after use contained in them

After reconstitution, the solution can easily be replaced with filled containers, without introducing air into the liquid path of the dispensing system.

[0022] If the containers that make up the reservoir for the respective solution are shaped as a sack in a sack, then the introduction of air bubbles into the dispensing system, in particular into the nozzles, is avoided. Thus, venting the container is also obsolete.

[0023] According to the development, at least one check valve is placed in the fluid line between the container and the micropump, in particular with pretension. The non-return valve blocks back fluid flow to the container tank. Due to the pre-tension, any excess pressure in the container tank is cushioned, and thus liquid dripping from the opening is prevented

About the outlet nozzle.

[0024] Furthermore, it is provided that at least one sealing device is arranged in the area of ​​the nozzle outlet opening, whereby it is possible to close the nozzle outlet opening, thereby avoiding reversal of the liquid meniscus as well as clogging of the nozzle outlet opening. The nozzles can be hermetically sealed, especially between measurements, and thus clogging of the nozzles will be eliminated.

[0025] The dispensing system comprises at least two dispensing units, wherein at least one container with the first solution is disposed in the first dispensing unit, especially chemiluminescent solution, which is connected to at least one micropump via a fluid line, and then with the nozzle, and in the second dispensing unit at least one container with another solution is placed, especially the enzyme solution, which is also connected to at least one micropump via a fluid line, and then with the nozzle, and optionally at least one container with additional solution is placed in the third dispensing unit, especially the washing solution, which is also connected to at least one micropump via a fluid line, and then with the nozzle. The parallel placement of at least two dispensing units has proved advantageous because in this way there are completely separate fluid paths in the dispensing system, which only after switching to the device can contact each other, and thus prevent premature reaction or contamination of one solution with another solution.

[0026] In each dispensing unit, the container may have a port connector attached, for connecting a standard Luer cone, at least one non-return valve is located in the fluid line between the container and the micropump, especially with pretension, thanks to, on the one hand, it is possible to easily replace containers from the remaining system for testing, especially the dispensing system, on the other hand, the container overpressure can be captured, to prevent drops from dripping onto the hole for applying samples of the microfluidic device, and the solution return flow to the container. [0027] The nozzles of the adjacent dispensing units are preferably arranged such that they enter nanoliter liter solution drops into the same apertures for applying the sample of the microfluidic device. In this connection, it has proved to be advantageous that the additional devices or means that direct the liquid stream to the sample application openings are outdated. It has proved advantageous to place the nozzle outlet end at a distance of 0.1 mm to 80 mm from the sample application opening microfluidic device, thanks to which the distance between the dispensing device and the microfluidic device has been covered, which enables contactless feeding

EP 2 988 871 81 of the respective solution into the opening for applying a sample of the renal fluid device. Thanks to tern, it is avoided that the solutions of the different metering units or nozzles infect each other.

[0029] In order to better understand the invention, it will be explained in more detail on the basis of the following figures. [0030] And so they show each time in a very simplified view:

Fig. 1 schematic view of the test system;

Fig. 2 shows the signal amplification through the optofluidic effect;

Fig. 3 a representation of the chemiluminescent signal in the measuring region of the riotrofluidic channel; Fig. 4 shows the shift of the chemiluminescent signal in the measuring region of the radical cell channel.

[0031] At the outset, it should be noted that in the various embodiments described, the same parts are provided with the same designators or the same designation of the components, wherein the disclosures contained in the entire description can be transferred, in the sense, to identical parts with with the same markers or the same designations of the components. Also the location data selected in the description, such as top, bottom, side, etc., refer to the figure directly described and depicted, and in the event of a change of position, they should be moved, according to the sense, to the new position. In addition, also individual features or combinations of features of the various embodiments shown and described can be self-contained, inventive or inventive solutions.

[0032] All data regarding ranges of values ​​in the present description should be understood to include any and all of their partial ranges, for example given 1 to 1O should be understood to include all partial ranges starting from the lower limit 1 and upper 1 O, i.e. all partial ranges start at the lower limit of 1 or more and end at the upper limit of 10 or less, for example 1 to 1.7, or 3.2 to 8.1 or 5.5 to 10.

[0033] The present invention describes a method of preparing a test system for use in in vitro diagnostics, especially in the field of point-of-care, and a test system.

[0034] Fig. 1 shows a schematic cross-sectional view of the test system 1 according to the invention, which comprises at least one dispensing system 2, a fluorescent device 3 and a photosensitive sensor 4 with a large number of photodetectors 5. The test system 1 is controlled by control modules.

[0035] Furthermore, a container 6 with a bag 7, a coupling 8, a check valve 9, a rotor tube 10 and a nozzle 11 of the dispensing system 2, which are connected to each other by a fluid line 12, is shown.

[0036] In addition to the opening 1 3 for applying the sample, the radiofluid device 3 comprises a radiophile fluid channel 14 with a measuring area with a plurality of test sections and optionally a reservoir 15. The fluid reflux device 3 is held by a receiving device 16 on which or in which the photosensitive sensor 4 is located .

[0037] By reacting the sample material transported in the rnikrofluidalny device 3 with the reagents from the dosing system 2 or via the reagents of the dosing system 2 itself, the test sections change the optical property or light emission based on the chemical reaction,

Relative biochemical, so that the photodetectors 5, assigned to each test section, detect the change in the incident light intensity.

[0038] The emission of light or light signal, as in the case of chemiluminescence or color change, can be based, for example, on a chemical or biochemical reaction, but also, for example during fluorescence or phosphorescence, on the supply of excitation energy from other sources. Preferably a chemiluminescent signal is detected.

[0039] In a possible embodiment, the dispensing system 2 is formed by a container 6 with a solution tank. The fluid line 12 connects the container 6 to the micropump 10 and, consequently, to the nozzle 11.

[0040] The dispensing system 2 is preferably formed of at least two containers 6, which are connected by one fluid line 12 to the micro pump 10 and at least two nozzles 11. The dispensing system 2 can be formed of several dispensing units 17, wherein the dispensing unit 17 comprises at least one container 6, fluid line 12, micropump 10 and nozzle 11. The nozzle 11 of the dispensing unit 17 can be closed by a sealing device 18.

[0041] The containers 6 serve as a reservoir for various solutions. In a possible embodiment, they are shaped as sacks 7 in a sack-like manner, which are known from infusion medicine, so that no air bubbles enter the fluid path of the dispensing system 2. Preferably, the chemiluminescent solution is contained in the bag 7, the enzyme solution in the next bag 7, and the washing solution in the additional bag 7. However, other solutions may also be present in the containers 6 which, for example, are necessary for the color reaction.

[0042] The sack-like bag 7 and fluid tube 12, which is shaped as a hose system, are connected to each other in the connector 8. In this connection, connector 8 is formed by a Luer connection. Sacks 7, sack-type, have a Luer cone at their lower end, which forms a liquid-tight and hermetic connection with fluid line 12. The sealing is achieved by the conical construction of the connecting parts, the so-called Luer cone. The container 6 with the tank can easily be replaced by a Luer connection without air bubbles.

[0043] Several such bags 7 can be combined in one housing and form their own structural unit. This construction unit can be easily disconnected via connector 8 from the remaining dispensing system 2, when, for example, a certain number of analyzes have been carried out using test system 1 and the volume of liquid contained in bags 7 has been used up. How often such a construction unit has been used, and thus when it needs to be replaced, this is assessed, for example, by an RFID system.

[0044] The micropump 10 allows the transfer of the smallest volumes. It contains injection fittings for the housing and pump chamber, piezo actuator and valves. Such micro-pumps 10 are known in the art, for example from Bartels-Mikrotechnik, or described in WO2009 / 059664 A1.

[0045] The fluid path of the solutions begins in the respective liquid reservoir in the container 6 and extends through the fluid conduit 12 to the micro pump 10 and ends in the respective nozzle 11. The micropump 10, in combination with the nozzle 11, creates a stream of nanolitre drops, which makes it possible to contactlessly and purposefully dispense a few microliters into the opening 13 for applying a sample of the microfluidic device 3. Nanolitre drops are droplets with volumes of 1 ni to 100 ni, preferably 2 ni to 10 ni, especially 5 ni.

[0046] In the area of ​​the outlet end or outlet opening, the nozzles 11 have a diameter smaller than 500 Pm. Preferably, the diameter of the nozzle 11 in this area is between 100 μm and 300 μm, especially 250 μm. [0047] In a preferred embodiment, a check valve 9 is arranged between the micropump 10 and the container 6 in a fluid path that blocks the back flow of the respective solution to the container tank

6. In addition, the check valve 9 can be positioned with a pre-tension, wherein the pre-pressure of the tank is equalized by the pre-tension, and thus liquid dripping from the nozzles 11 into the opening 13 for applying a sample of the microfluidic device 3.k is prevented. A development of the invention, a sealing device 18 which hermetically closes the nozzles 11 between measurements and thus eliminates clogging in the nozzles 11 can be located in

10 About the nozzle outlet 11. The sealing device 18 can be located separately for each nozzle 11 or only for selected nozzles 11. In an alternative embodiment, the sealing device 18 can also be formed in one piece for several nozzles 11, e.g. in the form of silicone pads .

[0049] Bag 7 with connector 8 for fluid line 12, with non-return valve 9 with inlet, and micro pump

10 in combination with the nozzle 11 form a dispensing unit 17 of the dispensing system 2.

[0050] Several of such dispensing units 17 may be arranged in parallel in the dispensing system 2. In a preferred embodiment, three dispensing units 17 are arranged side by side, the dispensing units 17 differing in solutions in the bag reservoir 7. The first bag 7 contains a solution which is responsible or necessary for a chemical or biochemical reaction, especially a chemiluminescent solution, in the second bag 7 another solution, preferably an enzyme solution, and in the third bag 7 an additional solution, especially a washing solution. To reach the same opening 13 for applying the sample of the microfluidic device 3, when three nozzles 11 arranged next to each other are used, both outer nozzles 11 are arranged at an acute angle relative to the central nozzles 11.

[0051] In an alternative embodiment, the dispensing system 2 may also contain only one dispensing unit 17, which consists of several bags 7 with one fluid line 12 each, which can be connected in one micropump 10 with several chambers for different solutions , and from there each solution is discharged in separated nozzles 11.

[0052] In order to ensure targeted delivery of the respective solution to the application opening 13 of the sample for microfluidic device 3, the outlet opening of the nozzle 11 is positioned from 0.1 mm to 80 mm from the application opening 13. Preferably at an interval of 1 mm to 50 mm, in particular at an interval of 2 mm to 20 mm.

[0053] The microfluidic device 3 includes at least one opening 13 for sample application, the microfluidic channel 14 and the reservoir 15. The microfluidic channel 14 has a length from 30 mm to 50 mm, width from 1 mm to 4 mm and a height from 10 μm to 200 μm , and can be made by injection molding.

Preferably, the channel has a length of 40 mm, a width of 2 mm and a height of 100 μm. A measuring area is located in the microfluidic channel 14, where molecules that have specific binding sites for specific target molecules in the biological sample are also immobilized. Based on the geometrical characteristics of the microfluidic channel 14, the fluid capillary movement occurs, from the sample application opening 13, through the microfluidic channel 14, up to the reservoir 15. After the sample has been delivered to the sample application opening 13, a pressure gradient is created in the channel 14 with the resulting

This is due to the capillary force towards the reservoir 15, whereby it is ensured that the sample is passed through the channel or the microfluidic system itself, and therefore that no means for generating the pressure difference or the flow movement is necessary. This is especially the so-called convection-driven hybridization, during which convection gradients are formed in channel 14, which, in addition to passing the sample through channel 14, also ensure that the sample material is directed towards the test sections of the measuring area (Squires TM et al., "Making it stick: convection, reaction and diffusion in surface-based biosensors", Nature Biotech, 26, 4, 2008). During the analysis, the analyte in the biological sample contacts the particles immobilized in the measurement area, thanks to which in each measurement area, in the presence of a suitable analyte, a chemical binding reaction occurs in the sample, which consequently leads to the addition of appropriate reagents. for chemical or biochemical reaction (e.g. chemiluminescence, color change, etc.) with light emission.

[0054] Further possibilities for forming the microfluidic device 3, to provide more light to the photosensitive sensor 4, is the shape of the bounding surface of the channel 14, opposite the photosensitive sensor 4, in an optically reflective manner; forming a concave cross-section in channel 14, which serves as a focusing lens; placing the light guiding structure in the microfluidic device 3; forming the microfluidic device 3 as an optical fiber plate. Details regarding the possible options for forming the microfluidic device 3 are contained in WO 2012/080339. In an alternative embodiment, the microfluidic channel 14 can also be filled by means of a pump, preferably a micropump. In this type of microfluidic device 3, no reservoir is required that causes a capillary effect.

[0055] The microfluidic device 3 is detachably arranged in or on the receiving device 16, so that the light-emitting side of the microfluidic device 3 is facing the photosensitive sensor 4. The photosensitive sensor 4 is housed in the base body, the individual photodetectors 5 being protected by a transparent cover layer. A large number of test sections are located in the measuring area of ​​the microfluidic device 3. During the analysis, the test sections are returned to the microfluidic volume, thanks to which the analyte capillary movement occurs in the sample application opening 13 when the sample is discharged. As a result, the analyte contacts the test sections in the measuring area.

[0056] The microfluidic device 3 is positioned on the receiving device 16 in a housing in which the test system 1 is at least partly contained, which is shaped in such a way that the microfluidic device 3 is seated in the fixed part of the receiving device 16 and is held in a manner fixed by a second movable and / or foldable part of the receiving device 16. It is also possible that a part of the receiving device 16 has a pre-tensioned element which, when the microfluidic device 3 is fitted, is compressed and thus fixes the microfluidic device 3 in the receiving device 16. The receiving device 16 can also be shaped like a drawer and by actuating the element, the drawers are pulled out of the housing, the microfluidic device 3 is applied and fixed and retracted, whereby the photosensitive sensor 4 with photodetectors 5 can already be placed in the drawer element.

[0057] The photosensitive sensor 4 with a large number of photodetectors 5 is arranged in such a way that, if the microfluidic device 3 is positioned and held in the receiving device 16, the test sections, in the measuring area of ​​the microfluidic channel 14 are adjacent its side emitting light through photodetectors 5 of photosensitive sensor 4. For this purpose, the receiving device 16 may have, for example, a stationary and movable, longitudinally movable, spring loaded spring bias, so that when the microfluidic device 3 is mounted, the movable part can be displaced in the longitudinal direction to facilitate the seating of the microfluidic device 3, and fixes it accordingly, after springback back to the holding position. In addition to the longitudinally displaceable configuration, a folding or latching mechanism can also be provided. It is also possible that at least in one part

There is a pressure means, e.g. a rubber or spring element, which, after seating, fixes the microfluidic device 3 described above.

(0058] The test system 1 is at least partly contained in the housing, the containers 6 of the dispensing system 2, which should be easily accessible, to allow replacement, can also be located outside the housing. The housing must provide light-tight closure of the microfluidic device 3 to the environment. A sample delivery device may be provided in the housing, which allows forwarding to the sample application hole 13 of the microfluidic device 3.

[0059] The receiving device 16 is shaped like a drawer and transports the microfluidic device 3 for priming in the housing. A light-sensitive sensor 4 with a large number of photodetectors 5 can also be placed in the drawer element. The sealing element closes the microfluidic device 3 and the photosensitive sensor 4 in relation to the surroundings. The sealing element can be formed, for example, through a tongue and groove connection. The sealing element can, however, also be formed by an elastically deformable element, for example foam material or a rubber gasket, whereby the closing of the drawer element, due to the compression of the sealing element caused by this, a light-tight closure of the inside of the measuring instrument with respect to the environment is created. In order to feed the biological sample, after pouring, into the sample application opening 13 of the microfluidic device 3, the drawer element is extended again.

In this way, the microfluidic device 3 can be embedded in the receiving device 16, and then the drawer element can be closed, where the sample material or sample chemistry is no longer in the microfluidic system, so that it is ensured that the test sections the measuring area triggers a chemical reaction. Only then, with the drawer element closed and the light-tight closure of the microfiber / uid device 3 reliable, priming takes place and the necessary measurements are carried out. Immediately after priming, the biological sample is applied, for which purpose the drawer element is first opened, and then, for at least one necessary measurement using a photosensitive sensor 4 with a large number of photodetectors 5, the drawer element is closed again in a light-tight manner . e (0061] A lighting device, e.g. LEDs, can also be arranged in the housing. Possible further embodiments of the receiving device 16, the lighting device and the implementation of the measuring system with a light-sensitive sensor 4 with a large number of photodetectors 5 are described in WO 2012/080339 and belong to the disclosure of the present invention.

[0062] In order to keep possible sources of defects to the lowest possible level, due to manual operations and personnel costs, and thus the overall costs of the analysis, the method which is carried out by means of the test system should be as significant as possible automated degree. After manual addition of the sample material, i.e. the biological sample being analyzed, the solutions necessary for the analysis are introduced into the opening 13 for applying the sample of the microfluidic device 3. However, this is problematic in this regard, because on the one hand, the volumes delivered can be kept as accurately as possible, or the order of delivery of the solutions used in the analysis is pre-determined. [0063] By means of the method according to the invention, the analysis is automated insofar as only the sample to be analyzed has to be applied manually.

[0064] By means of the inventive priming method of the system 1 for testing, changes in the light signal and / or light emission of chemical or biochemical reactions are measured and their results are used as parameters for controlling the filling of the dispensing system 2.

[0065] Necessary for each analysis, the microfluidic device 3 is embedded in the test system 1, especially on the receiving device 16 for this. The dosing system 2 must then be properly prepared to provide the reagents necessary for analysis with the correct volume, free from air bubbles and in the correct order. For this purpose, preparation or priming can be started via the control unit. Alternatively, preparation can also be started automatically as soon as it is recognized by the test system 1 that the microfluidic device 3 is located in the test system 1 and is also light-tight closed.

[0066] In its simplest embodiment, the solution in the container 6 is transported through the nozzle 11 of the dispensing system 2 to the opening 13 for applying the sample of the microfluidic device 3 by means of the micro pump 1O, and then it is further transported to the measuring area of ​​the microfluidic channel 14 of the microfluidic device 3. In the measuring area of ​​the microfluidic device 3, the light signal is measured by means of at least one light-sensitive sensor 4 with a large number of photodetectors 5, with changes in the light signal and / or light emission of chemical or biochemical reactions being measured. As soon as a light signal or a change in the light signal is detected, the micropump 10 is turned off. [0067] For the preparation or priming of the test system, by starting the micropump 10, the first solution from the first container 6, by means of which a chemical or biochemical reaction with the next solution is possible to emit light through a fluid path that contains at least one hose, a 1 O micro pump and a nozzle 11 are transported to the opening 13 for applying a sample of the microfluidic device 3. Simultaneously or subsequently, a further solution, preferably an enzyme solution, is transported from the second container 6 on the distal fluid path, also to the sample application opening 13. There, the first solution, especially the chemiluminescent solution, reacts with the next solution, preferably the enzyme solution, and a light signal is generated, especially a chemiluminescent signal, which can be detected as soon as the liquid is in the measuring area of ​​the microfluidic channel 14. In this way, it is ensured that both the first solution, especially the chemiluminescent solution, as well as the further solution, preferably the enzyme solution, is located in the nozzles 11 of the dosing system 2 and thus fills them for subsequent analysis and makes them ready.

EP 2 988 871 81 [0068] In an alternative embodiment, to be prepared, or priming of system 1 for testing, from the first bag 7 the first solution can be transported, especially chemiluminescent solution, via fluid line 12, through check valve 9, through the micro pump 1O to the first nozzle 11, and from there in the form of nanolitre drops it is supplied to the opening 13 for applying a sample of the microfluidic device 3, to transport from there by capillary action, through the measuring area, to tank 15. To this end, the micropump 10 is first started and a measurement is carried out in the measuring area of ​​the microfluidic channel 14. If during the measurement the signal changes or the predefined limit value is exceeded, which already occurs when a liquid meniscus occurs, the solution is in the microfluidic channel 14 and the micropump 10 is stopped because the first solution, especially the chemiluminescent solution, is in the outlet opening the first nozzle 11. In order to measure the change in the light signal, a light source, in particular an LED, may be arranged in system 1 for testing. Preferably, the first solution is transparent. Due to the shape of the microfluidic channel 14 and the refractive index of the liquid, through the effect of the optofluid lens, the photodetector 5 amplifies the light signal, which is sent by the light source when the transparent liquid is in the microfluidic channel 14. Due to the configuration of the test system 1 according to the invention, the signal strength in the microfluidic channel 14 is increased due to the lens effect caused by the flow of the first solution. This signal amplification is clearly visible and can therefore be easily detected, as shown in Fig. 2, wherein l (AU) is the change in the intensity of the light signal.

[0069] Then, a second micropump 10 is activated to transport a further solution, preferably an enzyme solution, from the second bag 7 via liquid line 12, check valve 9 and micropump 10, to the second nozzle 11, and from there supplied as nanolitre drops to hole 13 for applying a sample of the microfluidic device 3. Due to capillary action, the solution is also drawn into the measuring area of ​​the microfluidic device 3. Since the first solution already exists, especially the chemiluminescent solution, a chemical or biochemical reaction of both solutions occurs, with light emission, and a light signal is created, especially a chemiluminescent signal, which is measured in the measuring area. When a light signal is detected, the micropump 10 is stopped because a further solution, preferably an enzyme solution, is up to the outlet opening of the second nozzle 11.

[0070] In a preferred embodiment, the microfluidic channel 14 is still rinsed to on the one hand avoid premature reaction of the analyzed sample, which is later supplied, and on the other hand also prepare the nozzle 11 by means of a rinsing solution. To this end, the third micropump 10 is started, and an additional solution, especially the washing solution, is transported from the third bag 7, via the connector 8, to the fluid line 12, through the check valve 9 and the micropump 10 to the third nozzle 11, and from there nano-liter drops are supplied to the opening 13 for applying a sample of the microfluidic device 3. By adding an additional solution, especially a washing solution, an additional volume of liquid reaches the microfluidic channel 14 and the light signal, especially the chemiluminescent signal, is shifted. When this shift of the light signal is detected, especially the shift of the chemiluminescent signal, the third micropump 10 is turned off, because now also an additional solution, especially the washing solution, reaches the outlet end of the third nozzle 11.

[0071] Fig. 3 shows a typical distribution of the chemiluminescent signal in the measurement area of ​​the microfluidic channel 14, after the reaction of the chemiluminescent solution with the enzyme solution, wherein [pA] on the y axis is a photocurrent.

[0072] Fig. 4 shows the chemiluminescent signal shift after the washing solution is added.

[0073] At the latest after filling the third nozzle 11 up to the end of the outlet opening, the nozzles 11 can be closed by means of a sealing device 18. Of course, the nozzles 11 can be closed together or individually, also after each nozzle 11 has been filled, so that they are prevented clogging due to drying of the nozzles 11.

[0074] The nozzles 11 are now ready for the next analysis and volumes can be fed to the microfluidic device 3 with an accuracy of + 1 μ |

[0075] If the test system 1, in particular the dispensing system 2, is intended to be used in each case 10 μ | up to 100 μΐ in volume, 7 bags should have a capacity for about 50 tests in the tank, i.e. about 0.5 ml to 5 ml.

[0076] When performing subsequent analysis, the sample to be analyzed is first applied to the microfluidic device 3, wherein the sample can be fed by pipette or as drops directly into the sample application opening 13 or via a delivery device in the housing. The biological sample is then transported to the microfluidic channel 14, where the target molecules of the biological sample react via binding sites with molecules that are immobilized on the test segment of the measuring area. Then a further solution, preferably an enzyme solution, is automatically added and thus drawn into the microfluidic channel 14. By adding an additional solution, especially a washing solution, the excess of the further solution, especially the enzyme solution, is removed to avoid non-specific signals. Finally, the first solution, especially the chemiluminescent solution, is automatically delivered to the microfluidic device 3, which in reaction with the enzymes, specifically switched on in the test sections from the enzyme solution, produces a light signal, especially a chemiluminescent signal, which is measured by the light-sensitive sensor 4 the number of photodetectors 5, and by assigning to the test section in the measuring area it can be determined which analyte is contained in the sample. As already mentioned above, instead of the enzyme solution and chemiluminescent solution, other solutions that together give a color change can also be used.

[0077] The measurement of the light signal, especially the chemiluminescent signal, is preferably carried out by means of the light-sensitive sensor 4 with a plurality of photodetectors 5 arranged in the test system 1, but in an alternative embodiment it can also take place via an external sensor.

[0078] When performing the sample analysis, it is necessary to document the measurement result. Therefore, an identity or identification tag can be placed on the microfluidic device 3 and thus the direct assignment of the read signal waveform of the individual test sections can be transferred to the measurement report. In addition, different microfluidic devices 3 with different test sections can be used, so that, for example, a characteristic feature or configuration data of the test sections can also be stored in the identification tag. The tag is preferably read by means of a reader operating contactless on the test system 1 and, for example, it can be created from one or two-dimensional code, however it is also possible to be shaped as

RFID tag. This reader can be formed, for example, by means of an optical one- or two-dimensional detection sensor or RFID transmitting and receiving unit.

[0079] The embodiments show the possible variations of the implementation of the test system 1, it being noted here that the invention is not limited to the specifically illustrated variations thereof, but rather different combinations of individual variations between themselves and this variant possibility are also possible , based on the pattern of the technological procedure, thanks to the present invention depends on the skills of a specialist active in this field of technology.

[0080] For the sake of order, it should finally be noted that, for a better understanding of the structure of the test system 1, or its components, it is presented inconsistently with the scale and / or as enlarged and / or as reduced.

[0081] The task underlying the independent inventive solutions may result from the description.

List of markers [0082] test system dosing system microfluidic device light-sensitive sensor

5 photodetector container bag connector check valve

10 micropump nozzle fluid conduit opening for sample application microfluidic channel

15 tank receiving device dosing unit sealing device