Analysis system and procedure for performing an analysis

Abstract

An analysis system (10, 10', 10'', 10'''', 10'''', 10'''') for analyzing an analyte (13) which has the following features: at least one chamber (14) for processing the analyte (13), wherein the chamber (14) is formed by one or more polymer films and has a polymer with low thermal conductivity; a temperature-controllable element (15, 15''a, 15''c, 15''e) implemented as a wax plug valve or a heat-sensitive polymer valve; and a control layer (12l) comprising a heating device (12, 12a) for heating the at least one chamber (14) in order to carry out the processing of the analyte (13), and a further heating device (12c) coupled to the temperature controllable element (15, 15``a, 15``c, 15``e) and configured to control the temperature controllable element (15, 15``a, 15``c, 15``e; wherein the heating device (12, 12a) and the further heating device (12c) are formed by a common heating layer.

Description

[0001] One aspect of the invention relates to an analysis system comprising a control layer with a heating device, and to a corresponding method for controlling the analysis system. Preferred embodiments relate to an analysis cartridge or portable diagnostic systems comprising an analysis system.

[0002] Lab-on-a-Chip (LoC), lab-on-a-foil (lab-on-a-foil), micro-total analysis systems (µTAS), or bio-microelectromechanical systems (bio-MEMS) are common names for such systems that focus on the miniaturization of chemical and biological laboratory processes and experiments in microscale format, with the goal of improving the performance of the experiment and reducing its cost and time expenditure. These systems have in common that they handle liquids, as they generally consist of a set of microfluidic channels and / or cavities integrated onto a substrate. Depending on the application, these systems require a variety of other functionalities, such as mixing, heating, and filtering of samples and reagents, and detection of specific analytes or end products, in order to be able to replicate a corresponding laboratory scale.The various functionalities can be provided to the microfluidic system either through miniaturization and integration as on-chip components or can be kept separate as off-chip macroscale components. A macro-to-micro interface integrating the functionality into the microfluidic device offers advantages such as improved performance and sensitivity (direct contact with the sample) and a reduction of bulky off-chip components (important for wearable applications, PoC), but is associated with increased costs, as on-chip integration involves additional processing steps.

[0003] To avoid contamination risks, microfluidic systems are often disposable and require low material and manufacturing costs. Thus, plastics and polymers are preferred over the traditional MEMS material, silicone. Microinjection molding is the current state-of-the-art manufacturing technology for mass production of these systems. Despite the ability to produce millions of microfluidic plastic substrates with high throughput, the process presents difficulties in integrating functionalities. Injection-molded microfluidic systems are generally manufactured using solvent or thermal bonding, which is often incompatible with the electrodes and biomolecules to be integrated.The use of plastic films and roll-to-roll manufacturing is an attractive alternative mass production method, as the integration of such elements is straightforward and high-throughput using lamination-based processes and pressure-sensitive adhesive tapes. This allows different materials to be bonded together in a mix-and-match approach, allowing each component of the system to be manufactured under its optimal conditions.

[0004] Precise temperature control in microfluidic systems is often a crucial feature, as many biochemical reactions depend on specific temperatures. For example, cell growth and proliferation in a cell culture experiment require a constant temperature of 37°C. In a DNA hybridization experiment, the target and analyte strands only hybridize when incubated at 37°C. Furthermore, many DNA amplification protocols use specific temperatures to control the various steps in the amplification chain. Loop-mediated isothermal amplification (LAMP) is performed at a constant temperature of 60–65°C.Rolling circle amplification (RCA) requires a denaturation step at 95°C before elongation at 30°C. Finally, polymerase chain reaction (PCR) uses cycling between ~63°C, ~72°C, and ~95°C for exponential amplification of the template DNA. To achieve precise temperature control and high heating and cooling rates, integrating microscale heaters offers superior performance compared to external macroscale heaters due to direct thermal contact with the microfluidics.

[0005] We have previously demonstrated the detection of SNPs (single nucleotide polymorphisms) using melting curve analysis on DNA microarrays integrated using semitransparent copper grid heaters patterned on plastic film [1], [2], [3], [4]. The microheater was found to provide a robust and homogeneous temperature at the site of analysis, can be fabricated using a roll-to-roll process (low cost), and requires a power supply of 2 V (thus making it portable).

[0006] A conventional chip laboratory is in Fig. 9a shown. Fig. Figure 9a shows an analysis system 10* (also referred to as a chip lab) comprising a heating device 12*, e.g., a copper heater on a foil, and a chamber 14* for processing analytes. The chamber may, for example, be a DNA microarray, as shown by the enlargement of the chamber 14*. To transport the analytes, here the analytes containing DNA, into the chamber 14*, the layer of the chamber 14* may need to comprise one or more microfluidic channels 14mf*. The chamber 14* is formed in a layer adjacent to the layer comprising the heating device 12*. For example, the layer of the chamber 14* and the layer of the heating device 12* may be attached to each other using a structured double-sided adhesive foil 14a*.In addition, the layer of the chamber 14* may be covered by a film cover (made of PET / acrylic / PEN) 14c*, which is arranged on a side of the chamber opposite the layer comprising the heating device 12*.

[0007] The purpose of the analysis system 10* is to detect SNPs using melting curve analysis. Melting curve analysis is an assessment of the dissociation characteristics of double-stranded DNA during heating. As temperature increases, the double strand begins to split, leading to an increase in absorbance intensity, or hyperchromia. The temperature at which 50% of a DNA strand is denatured is known as the melting point. The information obtained can be used to interpret the presence and identity of the SNP.

[0008] In Fig. Figure 9b shows several melting curves. Typically, melting curves are plotted as diagrams showing the relationship between fluorescence and the temperature at which the fluorescence occurs. The melting curves shown belong to two groups: a first group, labeled 20m*, showing matches, and a second group, labeled 20mm*, showing mismatches. To detect such melting curves 20m* and 20mm*, the temperature of the analyte is raised quite precisely, with the analyte being monitored, for example, using an optical detector for its fluorescence or for another optical effect such as colorimetry or chemiluminescence.

[0009] As for the diagram of the Fig. As can be seen in Figure 9b, which illustrates the melting curve analysis, there is a need for fairly accurate temperature adjustment.

[0010] US 7 195 036 B2 shows a so-called microscale device and describes reactions within it. Furthermore, US 2006 / 0 036 348 A1 and US 2002 / 0 079 219 A1 also constitute further prior art.

[0011] State-of-the-art approaches, e.g., the approach of the analysis system 10* discussed above, often suffer from disadvantages related to the temperature constants within the chamber or the gradients across the lateral dimensions of the chamber. Another common disadvantage is related to the optical detector and its ability to accurately monitor the optical effect. Furthermore, the analysis system often incurs high manufacturing costs. Therefore, there is a need for an improved approach. Main aspect

[0012] An object of the present invention is to reduce the complexity and the resulting manufacturing costs.

[0013] This problem is solved by the subject matter of the independent patent claims.

[0014] An embodiment according to the main aspect already provides an analysis system for analyzing an analyte. The analysis system comprises at least one chamber for processing the analyte and a temperature-controllable element, such as a temperature-controllable valve, as well as a control layer. A heating device for heating the at least one chamber and a further heating device coupled to the temperature-controllable element are implemented in the control layer. The heating device for heating the at least one chamber has the purpose of supporting the processing in the chamber or enabling the processing, whereas the further heating device coupled to the temperature-controllable element is configured to control the temperature-controllable element by heating it. According to preferred embodiments, the heating device and the heating device are formed by a common heating layer.

[0015] Embodiments of this aspect are based on the realization that almost all or all elements can be implemented as temperature-controllable elements, eliminating the need to implement additional elements to control the chip. This is advantageous with regard to the complexity of the analysis system.

[0016] According to embodiments, at least one chamber can be configured for cell dissolution, DNA amplification, rapid DNA hybridization, and target detection, including a signal base layer and / or polymerase chain reaction. The temperature-controllable element can be, for example, a temperature-controllable valve, e.g., a valve having a wax plug configured to be either open or closed by controlling the ambient temperature.

[0017] Since the temperature-controllable elements and the various chambers for the various functions typically have different operating temperatures, the size of the heaters in the common layer may vary depending on further embodiments.

[0018] Another embodiment provides an analysis cartridge having pins that are connectable to a reading device that controls and / or evaluates the analysis and to an analysis system. Here, the pins are connected to the heating device and the further heating device of the control layer such that the analysis cartridge can be controlled externally.

[0019] Another embodiment provides a method for controlling an analysis system using the common heating layer. Side aspect

[0020] Embodiments belonging to a side aspect provide an analysis system for analyzing an analyte. The analysis system comprises a chamber and a heating device, e.g., a (copper) grid heater, integrated into a layer and having a two-dimensional geometry, e.g., a square. The layer is arranged adjacent to the chamber. The heating device is configured to release heat to the chamber according to a two-dimensional temperature distribution characteristic having a shape, e.g., an oval shape. A lateral shape and / or lateral size of the chamber are adapted to the shape and size of the two-dimensional temperature distribution characteristic. Consequently, the chamber may have a size comparable to the size of the temperature distribution characteristic and may have the same shape, e.g., an oval shape.

[0021] Embodiments according to this aspect are based on the finding that a heating device having a first geometry radiates heat toward the chamber having a different geometry due to the thermal conductivity at the flanks of the heating device. Based on this finding, the shape of the chamber is adapted to the shape of the temperature distribution characteristic and not to the shape of the heating device. Such a concept has advantageous effects on the temperature behavior and, above all, on the constants and gradients in the chamber.

[0022] According to embodiments, the aspect ratio of the two-dimensional temperature distribution characteristic is comparable or substantially identical to an aspect ratio of the chamber. Additionally or alternatively, the lateral shape of this chamber is comparable or substantially identical to the shape of the two-dimensional temperature distribution characteristic. In this context, "substantially" means that a mismatch of ±20% is acceptable, for example, if the aspect ratio of the temperature distribution characteristic is 20% smaller or larger than the chamber.

[0023] According to further embodiments, the size of the chamber and the temperature distribution characteristic can be comparable or essentially identical. A distinction can be made here for different cases. According to a first case, the chamber is 10% to 30% smaller than the temperature distribution characteristic. This case allows for a very small gradient from the center to the edge, resulting in an almost homogeneous distribution across the entire chamber. In a second case, the temperature distribution characteristic is identical to the chamber within a tolerance of ±20%. A third case, according to which the chamber is 10% to 30% larger than the temperature distribution characteristic, allows for a high gradient from the center to the edge. The gradient of the second case lies halfway between the first and second cases.

[0024] According to further embodiments, the chamber is configured for a polymerase chain reaction, wherein the chamber is larger than the two-dimensional temperature distribution characteristic, so that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid within the chamber. Based on this, not only can circulation of the fluid be achieved, but degassing of an adhesive material in the layer of the chamber can also be avoided or reduced.

[0025] Another embodiment provides an analysis system for analyzing an analyte. The system comprises a polymerase chain reaction chamber and a heater integrated into a layer adjacent to the chamber and having a two-dimensional geometry, wherein the heater is configured to deliver heat to the chamber according to a two-dimensional temperature distribution characteristic having a shape, wherein a lateral shape or lateral size of the chamber is selected such that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid within the chamber.

[0026] According to a further embodiment, the chamber is formed by one or more polymer films and / or comprises a polymer with low thermal conductivity. This has the advantage of achieving rapid cyclic temperature changes without increasing the complexity of the overall system. In this case, it should be noted that, according to preferred embodiments, the heating device is implemented as a thin-film heating device and arranged as a layer on the main surface of the chamber. Side aspect

[0027] One embodiment of this aspect of the invention provides an analysis system for analyzing an analyte. The analysis system comprises at least one chamber, a semitransparent heater, and an optical detector integrated into a layer adjacent to the chamber for detecting light coming from the chamber. The optical detector is configured to output a signal indicative of a transmission behavior through the chamber or an optical activity of the analyte.

[0028] Embodiments according to a further aspect are based on the finding that the integration of a chamber and a semi-transparent heating device on two different sides of the chamber enables an analysis system that offers high functionality, high detection accuracy, and low manufacturing costs. This concept also has advantages in terms of usability, since the analysis cartridge does not need to be positioned or aligned with respect to an external optical detector.

[0029] According to a further embodiment, a light source can be arranged on one side of the heating device, i.e., opposite the optical detector. Due to the fabrication of the detector and the light source directly on the microfluidic system, it is possible to increase sensitivity by reducing light loss through shorter light paths and a reduced number of light scattering interfaces. According to a further embodiment, the light source can be used in combination with an optical filter configured to convert a light source into a background with homogeneously distributed illumination. Here, a frequency change can also be performed such that a light frequency matches an absorption spectrum of the optical detector.

[0030] According to a further embodiment, the analysis system can be coupled to a processor or comprise a processor that enables the analysis of the light transmission behavior and / or the optical activity of the analyte. Light transmission through the chamber that exhibits low absorption indicates the case in which no reaction has occurred; light transmission through the chamber that exhibits high absorption indicates the case in which a reaction has occurred. Fluorescence, chemiluminescence, or colorimetry of the analyte indicates the case of an ongoing biochemical reaction.

[0031] Another embodiment provides a method for analyzing an analyte using the chip lab characterized above, wherein the signal indicative of the light transmission behavior and / or optical activity of the analyte is processed according to the discussion above.

[0032] All of the above embodiments according to the above aspects relate to an analysis system that uses heating devices. Based on this, it is clear that the technologies described above according to aspects are not compatible with each other, but must also be combined in a predetermined manner. Therefore, further embodiments provide an embodiment that has features of at least two aspects.

[0033] Another embodiment provides an analysis cartridge comprising an analysis system and pins that are to be connected to a reading device that controls and evaluates the analysis. Here, the pins are connected to the optical detector in such a way that external access to the detector is enabled.

[0034] Another embodiment provides a graphically displayable diagnostic system comprising an analysis system according to one of the above embodiments and a battery as the power supply. A small battery is sufficient here, since the heaters for heating the chambers and for controlling the elements are arranged directly next to the chamber or the temperature-controllable elements, so their power consumption is quite low.

[0035] Embodiments of the present invention will now be explained with reference to the accompanying drawings, in which: Fig. 1 shows a block diagram of a basic implementation of a side aspect analysis system; Fig. 2a-2f Diagrams for describing the background of the embodiments according to the aspect of Fig. 1 show; Fig. 3 shows a block diagram of a basic implementation of an analysis system according to the main aspect; Fig. 4 a block diagram of an improved implementation of the analysis system according to the aspect of Fig. 3 shows; Fig. 5 shows a block diagram of a basic implementation of an analysis system according to a further side aspect; Fig. 6 a block diagram of an improved implementation of the analysis system according to the aspect of Fig. 5 shows; Fig. 7a-7c show three different block diagrams illustrating the optical transmission behavior in the chamber for three different cases according to one aspect of the Fig. 5 show; Fig. 8a-8g show a further improved implementation of an analysis system according to an aspect of the Fig. 5 illustrate; and Fig. 9a, Fig. 9b refers to a conventional implementation of an analysis system.

[0036] Embodiments of the present invention will be explained in detail below with reference to the figures, wherein elements or structures that have a similar or identical function are provided with identical reference numerals. Thus, their descriptions are interchangeable and should be applicable to each other.

[0037] Fig. 1. View A shows a cross-sectional view of an analysis system 10 for analyzing an analyte 13 according to a secondary aspect, and view B shows a top view of the analysis system 10. The analysis system 10 has a chamber 14, which may be embedded in a layer comprising, for example, polymer films. This optional layer is designated by reference numeral 14l. Furthermore, the analysis system 10 has a heating device 12 integrated into a layer 12l located adjacent to the chamber 14 or adjacent to the layer 14l of the chamber 14.

[0038] The functionality of the analysis system 10 shown fulfills the functionality described above with reference to the conventional analysis system of Fig. 9 was described.

[0039] As with regard to Fig. As can be seen in Figure 1b, the heating device 12, e.g., a copper grid heater, has a two-dimensional geometry, e.g., a square. The square of the grid heater 12 is arranged parallel to the layer 12l, i.e., parallel to the layer 14l. The planar element is designed with flat, wide heat sink contact pads such that it radiates heat perpendicular to the plane and not to the sides. The heater 12 radiates or dissipates heat to the chamber 14 and its layer 14l. Therefore, a temperature distribution characteristic of the device 12 extends in the direction toward the chamber 14.

[0040] As a result of this two-dimensional geometry of the heating device 12, the shape of the temperature distribution characteristic designated by reference symbol 12tc is also two-dimensional. However, the shape of the two-dimensional temperature distribution characteristic 12tc differs from the two-dimensional geometry of the heating device 12. Here, the two-dimensional temperature distribution characteristic 12tc has an oval shape. The reason for this round shape 12tc is that the heat dissipation is concentrated in the center and decreases towards the boundary regions. In this context, it should be noted that the boundary or shape of the temperature distribution characteristic 12tc does not have a sharp transition; it is defined rather as a smooth transition. However, the transition bandwidth decreases disproportionately within this transition region, so that the shape of the temperature distribution characteristic 12tc can be clearly defined.

[0041] Based on this knowledge, the shape and size of the chamber 14 can be adapted to the two-dimensional temperature distribution characteristic 12tc to achieve the desired temperature constants in the chamber 14 or to achieve the desired temperature gradient from the center to the side of the chamber 14. In this embodiment, the chamber 14 has an oval shape, i.e., a shape comparable to the shape of the two-dimensional temperature distribution characteristic 12tc. Alternatively, the shape of the chamber 14 may be hexagonal or may have any other shape compared to the shape of the temperature distribution characteristic 12tc. Note that the above description regarding the shape of the chamber 14 refers to the two-dimensional shape in the region of the layer 14l, i.e., parallel to the temperature distribution characteristic 12tc.

[0042] Regarding the size of chamber 14, it should be noted that the size in the two-dimensional region essentially corresponds to the size of the two-dimensional temperature distribution characteristic 12tc. Here, "small" essentially means that the chamber can be smaller, e.g., 30% smaller, the same size, or larger, e.g., 30% larger, than the temperature distribution characteristic 12tc. The exact relationship between the two quantities depends on the desired temperature gradient.

[0043] There are various methods for comparing the two quantities. For example, the square of the temperature distribution characteristic 12tc can be compared with the projection of the chamber 14 to the plane 14l. Alternatively, the mismatch between the boundaries can be compared. To compare the similarity (mathematical similarity) of the two-dimensional temperature characteristic 12tc and the shape of the chamber 14 in the two-dimensional plane, the aspect ratio of the two comparable elements can be taken. For example, the two elements 14 and 12tc are similar, with the aspect ratio differing by a maximum factor of 1.5. Another method is to compare the orientation (x, y) of the boundaries of the two elements 14 and 12tc.

[0044] According to embodiments, the chamber 14 is formed by polymer films to enable precise thermal control and rapid thermal cycling. For example, a polymer with low thermal conductivity (compared to silicone, 40 cycles in six minutes) can be used.

[0045] According to further embodiments, the analysis system 10 is a film-based microfluidic PCR module to which a thin-film (copper grid) device for microPCR 12 has been applied. And PCR was carried out as in Fig. 2a. The high PCR efficiency is in Fig. 2b illustrates.

[0046] Fig. Figure 2b shows a graph of post-qPCR analysis results for four chips amplified using a foil-based µPCR system.

[0047] Since the three different steps in the PCR cycle (melting, extension, and denaturation) are each initiated by a specific temperature, it is essential to maintain the sample at the most uniform and precise temperature throughout the entire experiment. The PCR system uses the hot spot of a (copper grid) heater to define the shape of the PCR chamber. By varying the size of the microPCR chamber, chambers with either a constant temperature or different constant gradients can be achieved.

[0048] Fig. Figure 2c shows a hotspot 12tc' in a 4 × 6 mm grid heater 12'. The generated hotspot 12tc' in a 4 × 6 mm grid heater with a 30 µm line and a 300 µm space, as monitored with thermochromic liquid crystals (TLC), exhibits a bandwidth ranging from 60° to 65° C. Here, the hotspot 12tc' is oval.

[0049] Starting from this heating device 12', three different sizes of PCR chamber 14'a, 14'b and 14'c allow different temperature gradients from the center to the edge, as indicated by Fig. 2d is illustrated.

[0050] Fig. Figure 2d illustrates three designs (Design A, Design B, and Design C) corresponding to chambers 14'a, 14'b, and 14'c. The three designs A, B, and C differ from one another in terms of their size. All chambers 14'a, 14'b, and 14'c are oval, with each chamber having an inlet and an outlet. The chamber according to Design A has a width of 3.2 mm, i.e., 0.8 mm less than the width of the heater 12'. Design B has a width of chamber 14'b of 4 mm, i.e., equal to the width of the heater 12'. Design C provides a chamber 14'c that has a width of 4.8 mm, i.e., larger than the heater 12'.

[0051] For each design A, B and C, the resulting temperature gradient from the center to the edges is determined by Fig. 2e, where each chamber is filled with TLC, which has a temperature range of 60° to 65° C.

[0052] An analysis of the temperature distribution for the three different designs A, B, and C shows that the gradient for design A is quite small, 1° to 1.5° C from the center to the edge, whereas the gradient for design C is quite large, 3° to 4° C from the center to the edge. Design B has a gradient of 2° to 3° C from the center to the edge.

[0053] Exposure to a homogeneous or gentle gradient can have a beneficial effect depending on the type of PCR experiment. A homogeneous temperature can prevent nonspecific primer binding in some PCR experiments, whereas a multi-degree gradient can induce flow in the sample fluid between cold and warm regions and can have a stirring effect, promoting exchange between the chemical components in the fluid.

[0054] Challenges with microPCR include achieving precise thermal control and rapid thermal cycling with a polymer with low thermal conductivity. Similar results have recently been achieved with polymer-based systems, but these require rather complex setups, including liquid nitrogen reservoirs and air tubing, resulting in extensive instrumentation and increased costs. Using plastic films as a substrate for a microPCR chamber reduces the thermal mass of the low thermal conductivity material and allows for rapid adaptation to external forces such as applied heat or low ambient temperature. Thus, the film-based PCR chamber enables a high ramping and cooling rate without the need for additional external forces.For example, a cooling rate of ~4° C / sec (i.e. 95 to 63° C in ~8 seconds) was achieved using a simple convection setup (. Fig. 2f).

[0055] Furthermore, the heater used requires only a 2 V supply with a maximum current setting of 150 mA, meaning it could be powered by a small battery. This µPCR chip design enables a simple structure suitable for integration into portable molecular diagnostic systems.

[0056] With reference to Fig. 3 the main aspect is explained in detail. Fig. Figure 3 shows an analysis system 10' comprising the chamber 14 and another element 15, e.g., a valve, arranged adjacent to the chamber. The element 15 is a temperature-controllable element. Both elements 14 and 15 are arranged in the layer 14l. In a second layer 12l, arranged on the layer 14l, a plurality of heating devices, namely the heating device 12 and the heating device 12c, are arranged. The heating device 12 belongs to the chamber 14, wherein the design of the heating device, or more precisely, the design of the chamber 14, can be implemented as described above with reference to Fig. 1. The heating device 12c belongs to the temperature-controllable element 15, ie, the heating device 12c is arranged in the layer 12l such that the heating device 12c and the temperature-controllable element 15 are aligned with each other.

[0057] The temperature-controllable element 15, e.g., a valve with a wax plug, can be controlled by the heat generated using the heater 12c. Since the heater 12c is arranged in the same layer 12l that supports or controls the processing in the chamber 14, the layer 12a can be referred to as a control layer. Viewed from another perspective, this means that virtually every element of the analysis system 10' is controlled using heaters. Thus, it is advantageous that the heaters are formed by the same metal layer, reducing complexity, especially complexity related to manufacturing effort.

[0058] Since the chamber 14 typically requires a different temperature than the temperature-controllable element 15, the heaters 12 and 12c may differ in size, or generally in heating power.

[0059] According to further embodiments, the analysis system 10' may have a plurality of chambers; e.g., different heating devices would also be arranged in layer 12l for different purposes. This embodiment, which enables a fully integrated microfluidic sample-to-response system for nucleic acid testing, requires the manipulation of microvolumes of fluids in several sequential steps. For example, detection of a specific pathogen by its DNA in a patient's blood sample involves sorting the pathogen from the diverse components of whole blood (filtration / cell sorting mechanism), disrupting the cell membrane to extract DNA (cell lysis), amplifying the DNA, and detecting it. This requires diverse functionalities such as dissolving and mixing reagents, filtering solutions, and incubating samples.Managing the processes in sequential order generally also requires valve mechanisms and waste disposal. A variety of approaches to enabling each experimental step are possible. For example, a cell membrane (lysis) can be disrupted by exposing it to high temperatures, strong surfactants, or ultrasonic waves, all of which can be integrated into a microfluidic format. However, as previously described, integrating functionalities comes at the cost of increased manufacturing costs.

[0060] A schematic principle of such an analysis, which has several sequential steps, is shown by Fig. 4a. The implementation in a 10'' laboratory using a microfluidic section is shown by Fig. 4b and Fig. 4c illustrates. Fig. 4b shows a side view, where Fig. 4c shows a top view of the analysis system 10''. The analysis system 10'' comprises a plurality of reaction chambers 14, 14''b, 14''d, and 14''f connected by channels or, more precisely, by controllable channels that are controllable using temperature-controllable elements, which are discussed below. For controlling the temperature-controllable elements and for heating the chambers 14 to 14''f, a control layer 12''l comprising a plurality of heaters 12''a to 12''f is integrated into the analysis system 10''.

[0061] The elements 12''a to 12''f are thin-film copper heaters which are compatible with the heaters as described above with respect to the aspect of Fig. 1 are comparable. Element 12''b in combination with chamber 14''b enables cell dissolution, element 12''d in combination with chamber 14''d enables DNA amplification via PCR, and element 12''f in combination with chamber 14''f enables rapid target detection using DNA hybridization and / or melting curve analysis for SNP detection. Elements 12''a, 12''c, and 12''e provide heat to control the heat-induced valve mechanisms 15''a, 15''c, and 15''e. The elements 15''a with 15''c and 15''e, arranged between the chambers 14 and 14''b, 14''b and 14''d, and between 14''d and 14''f, serve to control the lateral flow between the various reaction chambers 14, 14''b, 14''d, and 14''f. Heat-induced valves 15''a to 15''e could be implemented as valves that hold a wax plug, e.g.an irreversible wax plug that can be melted using the heaters 12"b to 12"e. Alternatively, the heat-induced valves 15"a to 15"e may comprise expansion / compression elements made, for example, from heat-sensitive polymers.

[0062] The advantage of the thermal-control-only approach is that most functionalities can be created in a single process step, for example, a single copper layer, which greatly reduces the complexity of the manufacturing process. The solution could enable a truly cost-effective probe-to-response diagnostic system.

[0063] With reference to Fig. 5 a further aspect of the invention is discussed. Fig. 5 shows an analysis system 10''' comprising the chamber 14, e.g., in layer 14l, wherein the layer 12l comprises the one or more heating devices for the one or more chambers 14. In addition, the analysis system 10''' comprises an optical detector layer 20l comprising an optical detector 20 arranged and configured to detect light coming from the chamber 14. According to a preferred embodiment, the optical detection layer 20l is arranged opposite the heating layer 12l, i.e., such that the layer 14l comprising the one or more chambers 14 is arranged between the two layers 20l and 12l. The chamber 14 as well as the heating layer 12l can be implemented according to the discussion regarding the minor aspect and the main aspect.

[0064] The optical detector 20 may be implemented as discussed with respect to the minor aspect and the major aspect.

[0065] The optical detector 20 can be a simple optical detector comprising light-dependent resistors or a CCD element. The purpose of the detector 20 is to detect whether a chemical reaction has occurred, is currently in progress, or has not yet occurred.

[0066] According to further embodiments, the analysis system 10''' may comprise a light-emitting component 21, which is preferably arranged on the side of the heating device, ie opposite the optical detection layer 20l.

[0067] Whether the microfluidic system provides the platform for a cell cultivation experiment, detects a specific pathogen in a diagnostic test, or represents a microbioreactor for the production of a bioproduct, the result of the biochemical processes must be reported qualitatively and / or quantitatively to the macro world. Detection of analytes and generated products is generally achieved using a mass-sensitive, electrochemical, or optical device, making optical detection methods such as fluorescence, chemiluminescence, colorimetry, or absorbance by far the most common.In addition to reducing the need for bulky optical components in near-patient setups, the integration of on-chip light sources and a photodetector for analyte detection in microfluidic systems offers the following advantages: 1) Fabricating the detector and light source directly on the microfluidic system could enable increased sensitivity by reducing light loss through shorter light paths and a reduced number of light scattering interfaces. 2) The integrated miniaturized detectors can be structured into condensed arrays, enabling effective multiplexing.

[0068] With reference to Fig. 6 an improved embodiment is discussed. Fig. 6 shows an analysis system 10'''' comprising the optical detector layer 20l, the heat generation layer 12l and the layer 14l comprising the microfluidic cavity.

[0069] In this embodiment, the heating layer 12a is implemented as a layer stack comprising a film substrate 12f, a thin-film heater 12, and an encapsulation material 12e. Analogously, the optical layer 20l is implemented as a layer stack comprising a film substrate 20f, an electro-optically sensitive layer 20, integrated electrodes 20i, and an encapsulation material 20e. Regarding layer 14l, it should be noted that the chamber 14 is formed in an adhesive film 14f. Viewed from another point of view, this means that the semitransparent thin-film microheater is fabricated on a film substrate 1. The optical detection layer is fabricated on a film substrate 2. The two films are arranged in a superimposed manner over a microfluidic cavity by a double-sided adhesive film into which the contour of a microfluidic channel has been cut.The semitransparent heater allows the light source and detector to operate in a transmission-mode setup without the need for optical filters or lenses. The microfluidic cavity between the heater and the detector hosts the biochemical event. The biochemical process can occur either in liquid form or immobilized on either substrate 1 or 2.

[0070] Regarding the Fig. Figures 7a to 7c discuss the three possible events that can be detected using the detector. Fig. 7a to 7c an analysis system 10'''', wherein Fig. 7a shows a light path from the light source 21 to a detector 20 without any biochemical activity in the chamber 14. Fig. Figure 7b shows a light path of a biochemical activity with a blocking effect on light passing from the light source 21 to a detector 20. Fig. Figure 7c shows a light path following a chemical activity that produces a change in the wavelength of the light traveling from the light source 21 to a detector 20.

[0071] If no event has occurred in the chamber, a constant amount of light from the light source falls on the detector. If a biochemical event has occurred in the microfluidic cavity, two detection mechanisms can be used, depending on the experimental setup. Either the biochemical event generates a product that creates a steric hindrance for the light coming from the light source and arriving at the detector, as in Fig. 7b, whereby a reduction of the light reaching the detector is effected (absorbance measurement), or a change in the wavelength of the light arriving at the detector is effected, as in Fig. 7c (fluorescence, chemiluminescence or colorimetric measurement). In the case of an experiment such as Fig. 7c, optional optical thin-film filters can be produced by coating the substrate 1 ( Fig. 6), or the polymer material of the substrate 1 can be chosen to have optical filter characteristics to enable excitation of light of the correct wavelength for the selected marker.

[0072] The biochemical event could be a cell culture proliferating laterally and blocking light coming from the light source, a DNA hybridization assay, an immunoassay, or an amplification product from any of the previously described DNA amplification protocols. The hybridization assay or amplification product could be labeled either with markers for fluorescence, chemiluminescence, or colorimetric measurement, or with markers for absorbance measurement (polymer beads, gold nanoparticles, silver enhancement methods).

[0073] Fig. Figure 8a shows another analysis system 10''''', or in detail, an electro-optical measurement setup for a DNA hybridization detector using bead labeling. Here, the detector layer 20l' comprises an OSC layer (OSC = organic semiconductor layer) 20o, carbon fingers 20c, and a (PET) film 20f.

[0074] The light source layer 21' is arranged opposite the detector 20l'. Here, the light source 21 is formed by an LED 21l, which is configured, for example, to emit light in the range of 360 nm, and by an optical spacer 21s arranged between the layer 14l comprising the chambers 14' and the LED 21l.

[0075] Between the detector layer 20l' and the light source layer 21' is the layer 14l', which has the chambers 14 (fluidic chamber or cavity 14). This layer 14l' can also be referred to as the sensor layer 14l'.

[0076] Here, the sensor layer 14l' also includes a (PEN) film 14f, which is arranged on the spacer and encloses the cavity 14. The cavity 14 serves to accommodate the analyte 13 (here a DNA + beads) in such a way that it can be analyzed using the detector 20l'.

[0077] This detection is carried out with reference to Fig. 8b to 8g are discussed in detail.

[0078] Fig. Figure 8b shows schematically the detector of the Fig. 8a in two different cases, namely in the case of a detected match and in the case of a mismatch during DNA detection or the detection of hybridization events. By using biotin-labeled DNA targets and streptavitin-coated polymer beads of 1 µm in size, it is possible to detect different concentrations of a DNA target. In the case of a mismatch in chamber 14, no DNA and no target object has been matched to the DNA strand. As a result, the light emitted from the light source 21', which passes through the film 14f, is not absorbed in the chamber 14, so that the emitted light reaches the detector 20 in a virtually unabsorbed manner.

[0079] In the case of a match, the streptavitin-coated batch 15, which contains the complementary DNA strand, reacts with the DNA strand. This reaction can be detected due to the fact that the light emitted by the light source 21' reaches the detector 20, where a high level of light absorption has occurred.

[0080] Fig. Figure 8c shows the response to bead labeling based on four different cases in which samples A1, A2, and A3 were hybridized with a 1 µm biotinylated target, whereas B1, B2, and B3 were hybridized with 50 nm and C1, C2, and C3 with 2.5 nm. The two mismatch samples, Mismatch 1 & Mismatch 2, without bound beads, are used as control points. As can be seen, the different concentrations produce different optically detectable effects, allowing the detector 20 to distinguish between the concentrations.

[0081] An exemplary signal corresponding to three different concentrations and one mismatch is shown in Fig. 8d shown. Fig. Figure 8d shows the current response in the electro-optical detection layer to different DNA concentrations.

[0082] The above structure uses an organic semiconductor layer (OSC) 20o, e.g., FS102 with an absorption maximum at 400 nm, and an LED light source of ~360 nm, 21l. An optical fiber is configured in the structure to utilize the transmission and autofluorescence properties of the film layers 20f and 14f. The transmission spectra of the layers 20f and 14f are shown in Fig. 8e. For example, for the autofluorescence of the film layer (PEN) 14f, the excitation and emission spectra of PEN films peak at 337–356 nm and 425 nm, respectively. By choosing an optical source 21l to lie between these ranges, or generally within a range dependent on the film layer chosen, it is possible to form a film-based optical filter system in which the layer 14f converts the point-source LED light source (~360 nm) into a homogeneously distributed light source in the range of ~420 nm, which matches the absorption spectra of the OSC layer 20o (~400 nm). Whereas the film layer (PET) 20f allows transmission of the same without any alteration of the light.

[0083] Fig. Figure 8f shows the spectra indicating the autofluorescence property of the optical layer 20f and 14f due to the light source 21l.

[0084] Additionally, Fig.8g, the property of the optical film layer 14f to convert a point light source such as an LED 21l into a homogeneously illuminated background, which is more suitable for the bead-based light blocking structure implemented in the chamber 14. Whereas the film layer 20f on the chamber 14 allows the light to pass unhindered to the OSC layer 20o.

[0085] All of the systems discussed above, belonging to various aspects, are preferably fabricated entirely on film, for example, using roll-to-roll technology. Since the detector layer and the heating layer are arranged directly adjacent to the chamber, embodiments according to all aspects have low power consumption, allowing them to be battery-operated. Therefore, another embodiment provides a portable diagnostic system comprising one of the analysis systems discussed above and a battery as the power supply. Combining the elements into a microfluidic system would enable an analysis module for biological or chemical events that is extremely cost-effective and portable.

[0086] A further embodiment provides an analysis cartridge comprising an analysis system according to one or more of the aspects discussed above. The analysis cartridge system is, for example, in the shape of a credit card and has external pins via which the analysis system can be connected to a reading device in order to control / evaluate the analysis. For example, the pins are connected internally to the control layer, i.e., to the heating devices, or to the optical detector elements. The analysis cartridge can be a disposable element, in which the analysis electronics could be separate, i.e., integrated into the reading device. Conversely, this means that all actuators and sensors are integrated into the disposable element, while elements that can be reused are arranged externally.

[0087] Although the above embodiments were discussed primarily in the context of a device, it should be noted that further embodiments provide corresponding methods. Therefore, according to a further embodiment, a method for controlling an analysis system is provided.

[0088] This method begins in the embodiments discussed with respect to the main aspect and comprises the steps of heating at least one chamber to carry out the processing of the analyte and heating the temperature-controllable element to control the same. Another method relates to analyzing an analyte. This method begins in the embodiments according to a secondary aspect and comprises the main step of analyzing the signal of the optical detector, wherein a signal corresponding to a detected light transmission through the chamber having low absorption indicates a case according to which no reaction has occurred, wherein a signal corresponding to a light transmission through the chamber having high absorption indicates the case according to which a reaction has occurred.Another signal, belonging to fluorescence, chemiluminescence, colorimetry, or something else, indicates the occurrence of a chemical or biochemical reaction in progress.

[0089] It is understood that aspects discussed in connection with a device also represent a description of the corresponding method, wherein a block or component corresponds to a method step or a feature of a method step. Analogously, aspects described in connection with a method step also represent a description of the corresponding block, detail, or feature of the corresponding device. Some or all of the method steps may be performed by (or using) a hardware device such as a microprocessor, a programmable computer, or an electronic circuit. In some embodiments, some or more of the key method steps may be performed by such a device.

[0090] Depending on specific implementation requirements, embodiments of the invention may be implemented in hardware or software. The implementation may be performed using a digital storage medium, such as a floppy disk, DVD, Blu-ray Disc, CD, ROM, PROM, EPROM, EEPROM, or FLASH memory, on which electronically readable control signals are stored that interact (or can interact) with a programmable computer system to perform the respective method. Therefore, the digital storage medium may be computer-readable.

[0091] Some embodiments according to the invention comprise a data carrier comprising electronically readable control signals capable of interacting with a programmable computer system such that one of the methods described herein is carried out.

[0092] In general, embodiments of the present invention can be implemented as a computer program product with a program code, wherein the program code is effective to perform one of the methods when the computer program product is run on a computer. The program code can, for example, be stored on a machine-readable medium.

[0093] Other embodiments include the computer program for performing one of the methods described herein, wherein the computer program is stored on a machine-readable carrier.

[0094] In other words, an embodiment of the method according to the invention is thus a computer program which has a program code for carrying out one of the methods described herein when the computer program runs on a computer.

[0095] A further embodiment of the method according to the invention is thus a data carrier (or a digital storage medium or a computer-readable medium) on which the computer program for performing one of the methods described herein is recorded. The data carrier, the digital storage medium, or the recorded medium is typically tangible or non-volatile.

[0096] A further embodiment of the method according to the invention is thus a data stream or a sequence of signals that represents the computer program for carrying out one of the methods described herein. The data stream or the sequence of signals can be configured, for example, to be transferred via a data communication connection, for example, via the Internet.

[0097] A further embodiment comprises a processing device, for example a computer or a programmable logic device, which is configured or adapted to carry out one of the methods described herein.

[0098] A further embodiment comprises a computer on which the computer program for performing one of the methods described herein is installed.

[0099] A further embodiment according to the invention comprises a device or system configured to transmit a computer program for performing at least one of the methods described herein to a recipient (e.g., electronically or optically). The recipient may, for example, be a computer, a mobile device, a storage device, or the like. The device or system may, for example, comprise a file server for transmitting the computer program to the recipient.

[0100] In some embodiments, a programmable logic device (e.g., a field-programmable gate array) may be used to perform some or all of the functionality of the methods described herein. In some embodiments, a field-programmable gate array may interact with a microprocessor to perform any of the methods described herein. In general, the methods are preferably performed by any hardware device.

[0101] The above-described embodiments are merely illustrative of the principles of the present invention. It is understood that modifications and variations of the arrangements and details described herein will be apparent to others skilled in the art. Therefore, it is intended that the invention be limited only by the scope of the following claims and not by the specific details presented in the description and explanation of the embodiments herein. References [1] “Genotyping of single nucleotide polymorphisms by melting curve analysis using thin film semitransparent heaters integrated in a lab-on-foil system”; Anna Ohlander, Caterina Zilio, Tobias Hammerle, Sergey Zelenin, Gerhard Klink, Marcella Chiari, Karlheinz Bock and Aman Russom; Lab Chip, 2013, 13, 2075 2082. [2] “DNA melting curve analysis on semitransparent thin film microheater on flexible Lab-on-Foil substrate”; Anna Ohlander, Tobias Hammerle, Gerhard Klink, Caterina Zilio, Francesco Damin, Marchella Chiari, Aman Russom and Karlheinz Bock; Proceedings of 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, October 28 - November 1, 2012, Okinawa, Japan, pages 797-799. [3] “Foil-based DNA melting curve analysis platform for low-cost point-of-care molecular diagnostics”; Anna Ohlander, Stefanie Bauer, Harisha Ramachandraiah, Aman Russom and Karlheinz Bock; Proceedings of 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, 27-31. October 2013, Freiburg, Germany, 1770-1772. [4] Patent literature containing the grid heater concept: - WO 2014 / 064 040 A1 - DE 10 2014 221 734 A1 - DE 10 2015 202 353 B3;

Claims

[1] An analysis system (10, 10', 10'', 10''', 10'''', 10''''') for analyzing an analyte (13), having the following features: at least one chamber (14) for processing the analyte (13), wherein the chamber (14) is formed by one or more polymer films and comprises a polymer with low thermal conductivity; a temperature-controllable element (15, 15''a, 15''c, 15''e) implemented as a wax plug valve or heat-sensitive polymer valve; and a control layer (12l) comprising a heating device (12, 12a) for heating the at least one chamber (14) to carry out the processing of the analyte (13), and a further heating device (12c) coupled to the temperature-controllable element (15, 15``a, 15``c, 15``e) and configured to control the temperature-controllable element (15, 15``a, 15``c, 15``e); wherein the heating device (12, 12a) and the further heating device (12c) are formed by a common heating layer. [2] An analysis system (10, 10', 10'', 10''', 10'''', 10''''') according to claim 1, wherein the temperature-controllable element (15, 15''a, 15''c, 15''e) is configured to be controlled by the ambient temperature of the temperature-controllable element (15, 15''a, 15''c, 15''e). [3] An analysis system (10, 10', 10'', 10''', 10'''', 10''''') according to any one of the preceding claims, wherein the heating device (12, 12a) differs in size from the further heating device (12c). [4] An analysis system (10, 10', 10'', 10''', 10'''', 10'''') according to one of the preceding claims, wherein the at least one chamber (14) is configured for cell dissolution, DNA amplification, rapid DNA hybridization and target detection up to the signal base layer, cell isolation, DNA extraction, DNA detection and / or polymerase chain reaction. [5] An analysis system (10, 10', 10'', 10''', 10'''', 10'''') according to one of the preceding claims, wherein the heating device (12, 12', 12''a, 12''b, 12''c, 12''d, 12''e, 12''f) is integrated into a layer located adjacent to the at least one chamber (14) and having a two-dimensional geometry, wherein the heating device (12, 12', 12''a, 12''b, 12''c, 12''d, 12''e, 12''f) is configured to deliver heat to the chamber (14) according to a two-dimensional temperature distribution characteristic (12tc, 12tc'), wherein the two-dimensional temperature distribution characteristic (12tc, 12tc') has a shape, wherein a lateral shape and / or Size of the chamber (14) is adapted to the shape of the two-dimensional temperature distribution characteristic (12tc, 12tc'). [6] An analysis system (10, 10', 10'', 10''', 10'''', 10'''') according to any one of the preceding claims, wherein the analysis system further comprises an optical detector (20) integrated into a layer located adjacent to the at least one chamber (14) for detecting light coming from the chamber (14) and configured to output a signal indicative of a transmission behavior through the chamber (14) or indicative of an optical activity of the analyte (13). [7] Analysis cartridge comprising an analysis system (10, 10', 10'', 10''', 10'''', 10''''') according to any one of the preceding claims and pins to be connected to a reading device controlling the analysis, the pins being connected to the heating device (12, 12a) and the further heating device (12c) of the control layer. [8] A portable diagnostic system comprising an analysis system (10, 10', 10'', 10''', 10'''', 10''''') according to any one of the preceding claims and a battery as a power supply. [9] A method for controlling an analysis system (10, 10', 10'', 10''', 10'''', 10''''') for analyzing an analyte (13), the analysis system comprising: at least one chamber (14) for processing the analyte (13), wherein the chamber (14) is formed by one or more polymer films and comprises a polymer with low thermal conductivity; a temperature-controllable element (15, 15''a, 15''c, 15''e) implemented as a wax plug valve or heat-sensitive polymer valve; and a control layer (12l) comprising a heating device (12, 12a) for heating the at least one chamber (14) and a further heating device (12c) coupled to the temperature-controllable element (15, 15``a, 15``c, 15``e), wherein the heating device (12, 12a) and the further heating device (12c) are formed by a common heating layer, the method comprising the following steps: Heating the at least one chamber (14) to carry out the processing of the analyte (13); and Heating the temperature-controllable element (15, 15``a, 15``c, 15``e) to control it.

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