EP3538268A1

EP3538268A1 - Microfluidic device and method for analysing samples

Info

Publication number: EP3538268A1
Application number: EP17790781.3A
Authority: EP
Inventors: Bernd Faltin; Jochen Rupp; Juergen Steigert; Christian Dorrer; Karsten Seidl
Original assignee: Robert Bosch GmbH
Current assignee: Robert Bosch GmbH
Priority date: 2016-11-10
Filing date: 2017-10-26
Publication date: 2019-09-18
Also published as: US11376583B2; DE102016222035A1; KR20190084048A; KR102496821B1; WO2018086903A1; CA3043112A1; CN109906116A; US20190262832A1

Abstract

The invention relates to a microfluidic device (1) for analysing samples, comprising at least two fluidic pathways (2, 3) for receiving samples, and at least one capture area (11) for a detection unit (4) for measuring light, which is configured to capture light emitted from samples in the at least two fluidic pathways (2, 3), across the capture area (11).

Description

description

title

Microfluidic device and method for analyzing samples The invention relates to a microfluidic device and a method for analyzing samples.

From DE 10 2011 078 770 AI a microfluidic device is known. The microfluidic device comprises, in particular, fluidically interconnected channels. The microfluidic device is particularly suitable for transport and

Analysis of fluids suitable.

Furthermore, DE 10 2010 031 212 A1 discloses a multilayer system comprising a plurality of material layers, as well as a production method for producing such a multilayer system.

On this basis, a microfluidic device, with a method and with a system according to the features of the independent claims is described. By the features listed in the respective dependent claims advantageous refinements and improvements of the microfluidic device and the method are possible.

A microfluidic device for analyzing samples is presented, which has at least two fluidic paths for sample ingestion and at least one detection zone for a detection unit for measuring light, which is adapted to detect emitted light from samples in the at least two fluidic paths over the detection area , includes. The term "microfluidic" refers here in particular to the size of the microfluidic device The microfluidic device is characterized in that in the fluidic channels and chambers arranged therein physical phenomena that are generally associated with microtechnology are relevant, for example capillary effects , Effects (especially mechanical effects) associated with surface tensions of the fluid, effects such as thermophoresis and electrophoresis, which are usually dominant in microfluidics over gravitational effects, which may also be characterized by: The term "microfluidic" may also be characterized over the cross-sections within the device, w elche serve to guide the fluid. Typical are, for example, cross sections in the range of 100 μηη [microns] times 100 μηη up to 800 μηη times 800 μηη.

In particular, the microfluidic device may be a so-called "lab on a chip." Such a "lab on a chip" is intended and configured to perform biochemical processes. This means that functionalities of a macroscopic laboratory z. B. be integrated into a plastic substrate. The microfluidic device may e.g. As channels, reaction chambers, upstream reagents, valves, pumps and / or Aktuations-, detection and control units have. The microfluidic device can make it possible to process biochemical processes fully automatically. This can z. B. Tests on liquid samples are performed. Such tests can z. B. find application in medicine. The microfluidic device may also be referred to as a microfluidic cartridge. In particular, by introducing samples into the microfluidic device, biochemical processes can be carried out in the microfluidic device. The samples may also be admixed with additional substances which trigger, accelerate and / or facilitate biochemical reactions. An evaluation can be carried out via the detection unit by placing the microfluidic device in such a way that the detection unit can cover at least the detection range (ie, in particular, can receive light which originates from the detection range is sent out). The detection unit is not part of the microfluidic device. In particular, the microfluidic device can be filled with samples independently of the detection unit.

The microfluidic device is described by way of example by means of molecular diagnostic detection methods. Further areas of application of the microfluidic device may be, for example, in the field of immunology or clinical chemistry. In particular, the microfluidic device can be used for in vitro diagnostics.

The microfluidic device is preferably particularly adapted and designed to analyze nucleic acids. This may in particular include an analysis of DNA. In particular, the microfluidic device can facilitate the implementation of several, in particular also different analysis and / or detection methods. In particular, the microfluidic device may allow multiple analysis and / or detection methods to be performed simultaneously (or sequentially) and / or in combination. In particular, different analysis and / or detection methods can be carried out in different regions of the microfluidic device or in different regions of the fluidic paths.

The microfluidic device is configured and intended to receive samples in the fluidic paths. In this case, a sample can be divided into a plurality of the fluidic paths. Alternatively, several different samples may be taken separately in the different fluidic paths. The microfluidic device preferably comprises a microfluidic network (which is formed in particular by the fluidic paths). Particularly preferably, the microfluidic network is highly integrated. This means that a large functionality is possible in a small space. The microfluidic device preferably includes pumps, valves and control devices that are designed and arranged to route samples into and / or through the fluidic paths. In particular, the microfluidic device can be utilized by variously adjusting valves for various applications. The fluidic paths are preferably at least substantially separated from one another. This means that liquids and other substances in the individual fluidic paths do not come into contact with each other and / or mixed with each other, at least except for individual desired interfaces, where a thorough mixing is explicitly desired and is specifically produced. For example, a plurality of fluidic paths can be combined in a common reaction chamber (whereby the common reaction chamber then represents an interface between the individual fluidic paths). In that case, the fluidic paths are separated from each other except for openings in this common reaction chamber. Also, the fluidic paths can be completely separated from each other. In that case, there is no interface between different fluidic paths.

The fluidic paths are preferably still thermally separated so that the samples (parts) in the different fluidic paths may have different temperatures. Preferably, the temperature of the individual fluidic paths can be set individually. It is also preferred that radiation barriers are provided between the fluidic paths. In this way, it can be achieved in particular that an external radiation (eg for excitation purposes) can be coupled onto a fluidic path or locally limited.

The detection unit preferably comprises a sensor, in particular an optical sensor. The optical sensor is preferably configured to detect electromagnetic radiation (in particular light) and to convert it into an electronic signal. Preferably, the detection unit is configured to perform a time and location resolved measurement and to generate a time and location resolved measurement signal.

Furthermore, the detection unit preferably comprises a signal processing unit for the electronic processing of the recorded signal, as well as a signal reproduction unit for the visual representation of the recorded and processed signal. The signal processing unit is preferably designed as a computer (in particular comprising a computer processor). Preferably, the signal reproduction unit comprises a screen. Alternatively, it is preferable that the Detection unit has only one terminal that outputs an electronic signal that is suitable and intended for processing by a signal processing unit and a subsequent representation by a signal reproduction unit. For example, a computer can be connected to the port.

The detection unit is preferably set up to particularly well detect light of such a wavelength which is emitted by samples in the microfluidic device or from which it is expected that a sample typically investigated in the microfluidic device will emit this light, if a certain examination method is performed on the sample. The light may in particular be electromagnetic waves having a wavelength in the range of 150 to 900 nm [nanometers], in particular in the range of 300 to 700 nm. In particular, the light may be visible to human beings.

The light can be emitted from the samples as a result of biochemical processes. The samples can also be at least partially converted by biochemical processes into a substance which can emit light. Furthermore, substances that can emit light can also be released by biochemical processes. The light can be emitted in particular due to fluorescence. Preferably, the microfluidic device is set up such that an external excitation of the samples or of a substance formed or released by biochemical processes is possible. As a result, fluorescence can be particularly strongly excited and measured particularly well.

The biochemical processes may in particular be those processes which are usually carried out for an analysis of nucleic acids (or of DNA). Particularly suitable as analysis methods are (real time) amplification, melting curve analysis and microarray analysis.

Amplification is to be understood as meaning, in particular, the amplification of DNA by an enzyme (such as, for example, polymerase). In this case, fluorescent substances can be released. By measuring the fluorescent light, the degree of amplification can be determined. That is, a light intensity and / or a spatial Extension of a light signal can provide information about the degree of multiplication of the DNA. Preferably, the entire light emitted by a sample is detected, in particular over the entire extent of the sample. Alternatively, a representative section of the sample can be considered.

Representative means that only part of the sample is measured, and the measured values can be converted to the entire sample. In a representative section of a sample, a value measured for a non-scaling property preferably corresponds to an average of the entire sample. A non-scaling property does not depend on the amount of sample considered, such as. B. a density. In addition, a value measured at the representative portion of the sample, a scaling property (such as a mass) corresponding to the proportion of the portion of the total sample, is smaller than a value of that property measurable for the total sample.

If the light is measured in real time, the amplification can also be called a real-time amplification. By real-time amplification, a time course of the amplification can be detected. In particular, a quantitative multiplication of the DNA can be detected by means of (real-time amplification.) The real-time amplification can be, for example, a so-called "real-time polymerase chain reaction (real-time PCR)." The term "chain reaction C.chain reaction") refers to thereby, that a product of an amplification reaction can in turn be the starting material of a renewed amplification reaction.

Since the (real-time amplification with the described microfluidic device can take place in different separate fluidic paths, one can also speak of a multi-well (real-time) amplification The multi-well (real-time) amplification is preferably carried out in a multi-well chamber of the microfluidic device, The multi-well chamber preferably has a volume in the range from 5 μΙ to 50 μΙ [microliters], in particular in the range from 10 μΙ to 25 μΙ. The depressions preferably each have a volume in the pico or nanoliter range. In particular, melting curve analysis may involve heating of DNA. In this case, a fluorescent substance can be emitted at a characteristic temperature. The characteristic temperature may, for example, enable identification of a DNA segment. By measuring an intensity of fluorescent light against a (in particular continuously and controlled increased) temperature DNA can be identified. As with (real-time) amplification, it is preferred for melting curve analysis that light emitted from the sample can be detected over a full extent of the sample.

In particular, for microarray analysis, a microarray (i.e., an array with columns and rows of micrometer-sized structures) from a plurality of test cells may be used. In particular, several thousand test cells can be arranged in a microarray. The various test cells can be provided with different (known) test substances, in particular by using automated devices. By adding a sample to the microarray, hybridization (i.e., attachment) of components of the sample and various of the test substances can occur. This can lead to the emission of fluorescent light or to the release and / or formation of fluorescent substances. Also, the sample may be provided with a fluorescent agent such that the emission of fluorescent light indicates the presence of (a constituent) of the sample in a particular test cell. By measuring in which the test cells (i.e., by which of the test substances) fluorescent light emission occurs, constituents of the sample can be identified.

The microarray analysis can also be performed simultaneously with two or more samples. It can be z. B. a first sample are mixed with a first fluorescer and a second sample with a second fluorescer. If the light emitted by the first and second fluorescers differs in wavelength (ie, in color), both samples can be analyzed simultaneously by wavelength-selective measurement of the light. Such a microarray analysis with different samples can be used, for example, to compare two samples, in particular healthy and diseased cells. With the described microfluidic device, the comparison within a closed Systems take place (ie within the microfluidic device), whereby errors can be reduced.

In the microarray analysis, a spatially resolved measurement of the emitted light is preferred, in particular with a resolution which allows an evaluation of the individual test cells (that is to say that a pixel of a measuring signal is at least smaller than a test cell). The resolution is preferably at least so great that a test cell with at least ten pixels can be represented.

The analysis methods described are preferably carried out in combination with one another. For example, it is preferable first to carry out an amplification (in particular by quantitative measurement of a degree of proliferation in real time) and then to qualitatively examine the DNA present in an increased amount by means of microarray analysis or to identify the constituents of the DNA in a sample. In particular, the described analysis methods can be carried out with the described microfluidic device. This can be realized in particular simultaneously or chronologically immediately following one another. In particular, the microfluidic device is preferably designed in such a way that the detection area for the detection unit comprises a part of the microfluidic device which is set up and intended for the (parallel) execution of different analysis methods.

In particular, with the detection unit, it may be possible to detect an overall intensity of emitted light within the detection area (as required, for example, for (real time) amplification and melting curve analysis). Total intensity means that all the light that is emitted by the samples within the detection range is recorded in total. Thus, the light intensity is integrated over the detection area. It may also be possible to determine an intensity not for the entire coverage area, but only for a part of it. This can be particularly useful for detecting light emitted by one of the fluidic paths (or by a part of one of the fluidic paths, in particular, for example, by a reaction chamber). In that case it is z. B. possible to separate with the detection unit for two fluidic paths and simultaneously perform a (real-time) amplification, wherein a quantitative statement about a Degree of increase of DNA can be obtained in the respective fluidic paths. If, for example, a microarray analysis is then carried out (for example in a reaction chamber which can be filled with samples from two otherwise separate fluidic paths and which is likewise arranged within the detection range of the detection unit), the microarray analysis can be performed for the microarray analysis same

Detection unit can be used. For microarray analysis, spatially resolved detection of emitted light is preferred.

This means that rooms are preferably provided for different processes (eg for several separate (real-time) amplifications and for a microarray analysis). On the other hand, it is preferred that a spatially resolved detection of emitted light is possible. The detection unit can be designed cost-effective, in particular, if either only a particularly large detection range or only a particularly high spatial resolution is realized. A high spatial resolution over a large coverage area can only be realized at great expense. The microfluidic device described here offers the advantage of being designed in a particularly small space. Thus, within the detection range of the detection unit both z. B. several separate (real-time) amplifications, as well as a microarray analysis are performed, the spatial resolution is sufficient in particular for the microarray analysis. In particular, this can save costs for the detection unit (because no particularly high-resolution detection unit is needed) or it can be dispensed with the use of a plurality of detection units. The detection area preferably has an area of 200 to 2000 mm ²

[Square millimeters], in particular in the range of 500 to 1500 mm ² . For example, the detection area may be implemented as a 30 mm by 30 mm [millimeter] square. The position of the detection area on the microfluidic device may be variable. This means that by moving the microfluidic device relative to the detection unit of the

Detection area on the microfluidic device can be moved.

By dividing into several fluidic paths, biochemical processes can be carried out separately. In particular, reactions between different samples or components of a sample are suppressed and a more robust process control can be achieved. Such reactions between (constituents of) samples in different fluidic paths may also be referred to as cross reactions. Transverse reactions are undesirable and hindering in most applications. For example, at a high multiplex level of more than four primer pairs (for example, in particular from 6 to 60 primer pairs) or in parallel RNA and DNA amplification undesired reactions may occur. Also, a process time can be reduced by the division into the fluidic paths, in particular without loss of quality. This is the case, for example, in the embodiment in which RNA amplification takes place in a first of the fluidic paths and DNA amplification in a second of the fluidic paths. By dividing the samples into the different fluidic paths, a degree of complexity of a (chemical or biochemical) reaction, in particular a multiplex reaction, can be reduced. This can reduce reaction and / or process times.

Furthermore, it can be achieved by the division into the fluidic paths that different analyzes can be carried out at different times and / or that at least the evaluation of different analyzes can take place at different times. In particular, analyzes can be combined that take different lengths. Also, preliminary or intermediate results can be determined. The samples can be further processed after a preliminary or intermediate result. Optionally, several advance or

Intermediate results between different process steps can be determined.

Also reference and target molecules of an analysis can be processed separately. In particular, the reference and target molecules can be analyzed using only one wavelength of (fluorescent) light.

In a preferred embodiment of the microfluidic device, the detection unit is designed with a camera. The camera can be embodied, for example, as a CCD or CMOS camera or have a CCD or CMOS chip. The camera is preferably set up to detect the detection range of the microfluidic device spatially resolved and over a large area (ie to detect light which is emitted from the detection range, in particular of samples in the microfluidic device). The camera is particularly preferably designed for the detection of fluorescence, chemiluminescence and / or bioluminescence. The camera is preferably sensitive in particular to electromagnetic radiation (ie, in particular to light) having wavelengths in the range of 150 to 900 nm [nanometers], in particular in the range of 300 to 700 nm.

In a further preferred embodiment, the microfluidic device has a coupling-in region for coupling an excitation emitted by an excitation device into the samples.

The excitation device is preferably a source of radiation, in particular electromagnetic radiation. Preferably, the excitation device is adapted to emit electromagnetic radiation having a wavelength in the range of 150 to 900 nm [nanometers].

The excitation device is preferably a heat source. Particularly preferably, the excitation device comprises a laser. The laser is preferably set up for fluorescence excitation (in particular within the samples). The excitation device is not part of the microfluidic device.

An effect of the excitation device on the samples is possible over the (entire) coupling region, whereby not all points of the coupling-in area have to be acted upon at the same time. The coupling-in area is the area in which an impact is possible. Preferably, the coupling-in region of the excitation device overlaps with the detection region of the detection unit. In particular, it is preferred that the coupling-in region of the excitation device and the detection region of the detection unit are (completely) congruent. In one embodiment, the excitation device is adapted to act with electromagnetic radiation of one or more (discrete) wavelengths on the samples in the microfluidic device, or to couple such electromagnetic radiation into the samples in the microfluidic device. By such action or coupling in particular fluorescence can be generated within the samples.

Particularly in the case of (real-time) amplification, multi-well real-time amplification and melting curve analysis, excitation is preferably carried out, ie. H. Excitation of the sample. A microarray analysis is preferably also carried out under excitation. Alternatively, however, microarray analysis may also be autofluorescent (i.e., without external excitation).

It is also preferable for the excitation to take place via an excitation light and in particular via a filter system (in particular for setting an excitation wavelength).

In a further preferred embodiment of the microfluidic device, at least one chamber is provided in each of the fluidic paths for receiving at least a portion of the sample.

The chamber can be, for example, a process or reaction chamber for carrying out a (bio) chemical reaction, an amplification chamber for performing a (real-time) amplification, a detection chamber for measuring (in particular fluorescence and in particular by means of the detection unit), a mixing chamber for Mixing a sample with a (test) substance and / or a storage chamber for (inter) storing a sample, a reaction (between) product or a (test) substance act. Also, a chamber can be used for several different purposes at the same time or in succession. Each of the chambers may be divided into a plurality of cells to form a microarray for microarray analysis.

In one embodiment, an amplification chamber (ie, a chamber in which amplification can take place) for DNA and / or RNA amplification is so arranged that the entire amplification chamber or at least a representative part thereof (eg with a size of 2 mm by 2 mm [millimeters]) is within the detection range of the detection unit. Thus, a melting curve analysis or a (real-time) amplification can be performed and recorded directly in the amplification chamber. In particular, it is preferred that the at least two fluidic paths are arranged in such a way that amplification chambers of all the paths are at the same time arranged in the detection range of the detection unit.

Also preferred is the embodiment in which a detection chamber is (fluidically) connected to an amplification chamber, so that a sample from the amplification chamber can be transferred into the detection chamber. In particular, it is preferred that the detection chamber lies within the detection range of the detection device.

It is preferred that the chambers can be tempered in particular between 25 ° C and 100 ° C, preferably even between 15 ° C and 100 ° C. In particular, it is preferred that the chambers can be individually brought to independent temperatures. Cooling and / or heating means are preferably provided for cooling and / or heating the chambers. The heating means may be, for example, heating wires for generating resistive heat or radiation sources for generating radiative heat. The coolants can be, for example, cooling lines for a cooling medium.

The chambers are preferably arranged within a common plane. Alternatively, it is preferred that the chambers are arranged in a plurality of planes, which are arranged in particular parallel to the detection region (or a surface of the microfluidic device).

In a further preferred embodiment, the microfluidic device further comprises an end chamber, which is connected to the at least two fluidic paths. Also, the end chamber may be divided into a plurality of cells to form a microarray for microarray analysis. The end chamber can be, for example, a process or reaction chamber for carrying out a (bio) chemical reaction, an amplification chamber for performing a (real-time) amplification, a detection chamber for measuring (in particular fluorescence and in particular by means of the detection unit), a mixing chamber for Mixing a sample with a (test) substance and / or a storage chamber for (inter) storing a sample, a reaction (between) product or a (test) substance act. Also, the end chamber can be used for several different purposes at the same time or in succession. It is also preferred that the end chamber is designed as the multi-well chamber described further above.

The end chamber is preferably connected to the at least two fluidic paths in such a way that the samples can be guided from the fluidic paths into the end chamber and mixed there. It is preferred that, with the exception of an indirect connection of the fluidic paths via the end chamber, there is no connection between the fluidic paths (ie that they are separated from one another).

In the end chamber, a microarray is preferably provided. Preferably, the end chamber lies completely within the detection range of the detection unit. Alternatively, at least the microarray is (completely) within the detection range.

In one embodiment, a (real-time) amplification (eg in the form of a real-time PCR) precedes a microarray analysis. In this case, the (real-time) amplification can be carried out in one or more of the fluidic paths (in particular separately from one another), while the microarray analysis is preferably carried out in the end chamber. For this purpose, the reaction product or the reaction products of the (real-time) amplification can be mixed with a hybridization buffer, pumped into the end chamber (which serves here as analysis chamber) and hybridized in the end chamber. This approach has the advantage that both (quantitative) information can be generated via the (real-time) amplification, as well as via the detection on the microarray multiplex detection can be performed. The microfluidic conversion in the fluidic paths that terminate in the end chamber can enable a surface-density conversion in a plurality of separate regions (in particular within different chambers), in particular within the detection range of the detection unit.

As a further aspect, a method for analyzing samples using a microfluidic device as described, comprising at least the analysis of nucleic acids according to at least two different analysis methods, wherein in each case different analysis methods are performed in different fluidic paths of the device.

The particular advantages and design features of the microfluidic device described above are applicable to the method described and transferable, and vice versa.

The fact that the method comprises at least the analysis of nucleic acids according to at least two different analysis methods means that at least two analysis methods are carried out, which can be carried out successively or (at least partially) simultaneously. Also, the at least two different analysis methods may be performed at different locations or (at least in the case of the sequential analysis methods) at a single location of the microfluidic device. Furthermore, the at least two different analysis methods can be carried out with the same sample (or with the same part of a sample) or else with different samples. In the latter case, it is preferable that there is an interaction between the different samples. This may mean, for example, that two samples are first treated separately from each other according to an analysis method, and then a joint analysis is carried out, for which purpose the two samples are mixed.

Preferably, a sample is divided into a plurality of the fluidic paths. Alternatively, it is preferable to separately receive a plurality of different samples in the different fluidic paths. It is also preferable to divide a first sample onto a first plurality of the fluidic paths and a second sample in a further one Fluidic path or distribute on a second plurality of fluidic paths.

By designing the microfluidic device with two paths, it is possible to perform two analysis methods, with two different samples, with a microfluidic device.

In a preferred embodiment of the method, the at least two analysis methods are selected from the following group:

- (real time) amplification,

- Endpoint amplification

- melting curve analysis and

- Microarray analysis.

The (real-time) amplification is preferably a polymerase chain reaction (PCR). By endpoint amplification is meant, in particular, an amplification in which a measurement of the amplification takes place in a late phase or at the end point of the amplification. As endpoint amplification, an endpoint PC R is preferred.

In a further preferred embodiment of the method, at least part of the sample is alternately pumped between at least two chambers having different temperatures, so that the part of the sample undergoes a thermal cycle.

It is particularly preferred if at least two samples are brought together in an end chamber, which is connected to the at least two fluidic paths of the device. Particularly preferably, the at least two samples are mixed together in the end chamber. It is further preferred if a microarray analysis with the mixed samples is carried out in the end chamber.

In such a method, processes with the pure, non-mixed samples can be carried out in the two paths. In addition, a final analysis of the mixed sample can be carried out in the final chamber. In this embodiment, the reaction mixture (ie the samples to which additional substances have possibly been added) are preferably pumped back and forth between two (or even more) different chambers, ie cyclically from a first chamber into a second chamber, from the second chamber in the first chamber, etc. In this case, the reaction mixture can be thermally cycled by keeping the first and the second chamber at different temperatures. This approach can allow for particularly rapid thermal cycling because only the temperature of the reaction mixture is changed while the environment, ie, the microfluidic device, and particularly the reaction chambers, can be maintained at (various) constant temperatures. Would the

If the reaction mixture is thermally cycled in a single reaction chamber, not only the temperature of the reaction mixture but also the temperature of the reaction chamber (i.e., in particular walls of the reaction chamber) would also have to be cycled. This can mean a greater amount of time.

The chambers involved in the thermal cycling are preferably arranged within the detection range of the detection unit. As a result, an (intermediate) measurement can take place via the detection unit between individual cycles. An (intermediate) measurement can also take place within a read cycle between two thermal cycles. Preferably, the chamber in which the

Reaction mixture is in the readout cycle, externally excited at least for a part of the duration of the readout cycle (eg by a laser). Thereby, the emission of a fluorescence signal can be excited, which can be detected by the detection unit.

As a further aspect, a system is presented, comprising a microfluidic device as described, a detection unit and preferably also an excitation device. The particular advantages and design features of the microfluidic device and the method described above are applicable to the system described and transferable. The invention and the technical environment will be explained in more detail with reference to FIGS. The figures show particularly preferred embodiments, to which the invention is not limited. In particular, it should be noted that the figures and in particular the illustrated proportions are only schematic. They show schematically:

Fig. 1: a microfluidic device for the analysis of samples and

2: a system comprising in particular the microfluidic device from FIG. 1.

1 shows a sectional view of a microfluidic device 1 for the analysis of samples. The microfluidic device 1 comprises a first fluidic path 2 and a second fluidic path 3 for receiving samples. In addition, the microfluidic device 1 comprises a detection area 11 for a detection unit 4 (shown in FIG. 2) for measuring light, which is set up to detect emitted light from samples in the two fluidic paths 2, 3. The detection area 11 is shown by dashed lines. In the first fluidic path 2, a first chamber 8 is provided for receiving at least a portion of the sample. In the second fluidic path 3, a second chamber 9 is provided for receiving at least a portion of the sample.

The microfluidic device 1 furthermore has a coupling-in region 12 for coupling an excitation emitted by an excitation device 6 (shown in FIG. 2) into the samples. The coupling-in area 12 is shown by dotted lines. The coupling-in region 12 partially overlaps the detection region 11.

In addition, the microfluidic device 1 has an end chamber 10, which is connected both to the first fluidic path 2 and to the second fluidic path 3.

The microfluidic device 1 can be used, for example, in the first two different samples, each containing DNA to be analyzed initially process fluidic path 2 and the second fluidic path 3 separately from each other. For example, a (real-time) amplification can be carried out in the first chamber 8 and in the second chamber 9. This can be detected quantitatively by the detection unit 4 because the detection area 11 completely encloses the first chamber 8 and the second chamber 9. Subsequently, the samples in the end chamber 10 can be further processed. For this purpose, for example, a microarray may be provided in the end chamber 10. A microarray analysis in the end chamber 10 can also be performed with the detection unit 4, because the end chamber 10 is located within the detection area 11. About the excitation device 6, the samples can be excited. In this case, excitation in the first chamber 8, in the second chamber 9 and in the end chamber 10 is in each case possible in part because the coupling region 11 partially encloses the respective chambers.

FIG. 2 shows a system 13 comprising the microfluidic device 1 from FIG. 1, a detection unit 4 and an excitation device 6. The detection unit 4 is designed with a camera 5. The excitation device 6 is designed with a laser 7.

By means of the detection unit 4 or by means of the camera 5, it is possible to detect light emitted by samples within the detection area 11 of the microfluidic device. Dotted lines indicate which area the camera 5 can cover. The detection area 11 is formed on the surface of the microfluidic device 1 between the dotted lines.

By means of the excitation device 6 or by means of the laser 7, it is possible to act on the microfluidic device 1 or on a sample located therein. This is indicated by a dashed line. Preferably, the excitation device 6 can be adjusted in such a way that excitation is possible at any point of the (in particular of the entire) detection region 11. All of the areas of the surface of the microfluidic device 1 which can be reached by the laser 7 together form the coupling-in region 12 (only shown in FIG. 1). The excitation by the excitation device 6 can take place for a limited time.

Claims

claims

Microfluidic device (1) for analyzing samples comprising at least two fluidic paths (2, 3) for sample taking and at least one detection area (11) for a detection unit (4) for measuring light, which is adapted to emit emitted light from samples in the at least two fluidic paths (2, 3) via the detection area (11) to detect.

2. Microfluidic device (1) according to claim 1, wherein the detection unit (4) with a camera (5) is executed.

3. Microfluidic device (1) according to one of the preceding claims, further comprising a coupling-in region (12) for coupling a, by an excitation device (6) emitted excitation in the samples.

4. Microfluidic device (1) according to one of the preceding claims, wherein in each of the fluidic paths (2, 3) at least one chamber (8, 9) is provided for receiving at least a portion of the sample.

The microfluidic device (1) according to any one of the preceding claims, further comprising an end chamber (10) connected to the at least two fluidic paths (2, 3).

6. A method for analyzing samples using a microfluidic device (1) according to one of the preceding claims, comprising at least the analysis of nucleic acids according to at least two different analysis methods, each different analysis methods in different fluidic paths (2, 3) of the device (1 ) be performed.

7. The method of claim 6, wherein the at least two analysis methods are selected from the following group:

- (real time) amplification,

- endpoint amplification,

- melting curve analysis and

- Microarray analysis.

8. The method of claim 6 or 7, wherein at least a portion of the sample is alternately pumped between at least two chambers (8, 9) with different temperatures, so that the part of the sample undergoes a thermal cycle.

9. The method according to any one of claims 6 to 8, wherein, at least two samples in an end chamber (10) are merged, which is connected to the at least two fluidic paths (2, 3) of the device (1).

10. System (13) comprising a microfluidic device (1) according to one of claims 1 to 5, a detection unit (4) and preferably also an excitation device (6).