DE102016211356B4 - Analysis system and method for performing an analysis - Google Patents

Abstract

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 perform 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

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.

Lab-on-a-chip (LoC), lab-on-a-foil, micro-total-analysis-systems (µTAS) or bio-microelectromechanical systems (bio-MEMS) are common names for such systems that deal with the miniaturization of chemical and biological laboratory processes and experiments in microscale format with the aim of improving the performance of the experiment and reducing its cost and time expenditure. What these systems have in common is that they deal with liquids, as they generally consist of a set of microfluidic channels and/or cavities integrated on 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 analogously replicate a corresponding laboratory scale. The different 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, whereby a macro-to-micro interface integrating the functionality into the microfluidic device brings 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 comes at the expense of increased costs as on-chip integration involves additional processing steps.

To avoid contamination risks, microfluidic systems are often disposable single-use systems with the requirement of low material and manufacturing costs, and thus plastics and polymers are preferred over the classic 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. The injection molded microfluidic system is generally manufactured by solvent or thermal bonding, which is often incompatible with electrodes and biomolecules to be integrated. Using plastic films and roll-to-roll manufacturing is an attractive alternative mass production method, since integration of such elements is readily achieved with high throughput through the use of lamination-based processes and pressure-sensitive adhesive tapes. This allows different materials to be bonded together in a mix-and-match approach and each component of the system can be manufactured under its optimal conditions.

Precise temperature control in microfluidic systems is often an essential functionality, as many biochemical reactions depend on specific temperatures. For example, cell growth and proliferation in a cell cultivation experiment require a constant 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 carried out at a constant 60-65° C. Rolling circle amplification (RCA) requires a denaturation step at 95°C before extension at 30°C, and finally, polymerase chain reaction (PCR) uses cycling between ~63°C, ~72°C, and ~95°C for exponential amplification of the template DNA. To obtain precise temperature control and high heating and cooling rates, integration of microscale heaters has better performance compared to external macroscale heaters due to direct thermal contact with the microfluidics.

We have previously demonstrated the detection of SNPs (single nucleotide polymorphisms) by 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 (therefore it is portable).

A conventional chip laboratory is in 9a shown. 9a shows an analysis system 10* (also referred to as a chip lab) comprising a heater 12*, e.g. a copper heater on a foil, and a chamber 14* for processing analytes. The chamber may be, for example, a DNA microarray, as shown by the enlargement of the chamber 14*. To transport the analytes, here the analytes comprising 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 that is adjacent to the layer comprising the heater 12*. For example, the layer of the chamber 14* and the layer of the heater 12* may be attached to one another by means of 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*.

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

In 9b several melting curves are shown. Usually, the melting curves are plotted as diagrams showing the dependence between a fluorescence and a temperature at which the fluorescence takes place. The melting curves shown belong to two groups, namely a first group marked 20m* showing matches, and a second group marked 20mm* showing mismatches. To detect such melting curves 20m* and 20mm*, the temperature of the analyte is raised quite precisely, the analyte being monitored for its fluorescence, for example using an optical detector, or for another optical effect such as colorimetry or chemiluminescence.

As with the diagram of the 9b As can be seen in Figure 1, which illustrates the melting curve analysis, there is a need for fairly precise temperature adjustment.

The US 7 195 036 B2 shows a so-called microscale device and describes reactions in it. In addition, the US 2006 / 0 036 348 A1 and US 2002 / 0 079 219 A1 further state of the art.

Prior art approaches, such as the analysis system 10* approach discussed above, often suffer from disadvantages related to constant temperature within the chamber or 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

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

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

An embodiment according to the main aspect already provides an analysis system for analyzing an analyte. The analysis system has at least one chamber for processing the analyte and a temperature-controllable element such as a temperature-controllable valve and 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.

Embodiments of this aspect are based on the realization that almost all or all elements can be implemented as temperature-controllable elements, so that no further elements need to be implemented to control the chip lab. This is advantageous with regard to the complexity of the analysis system.

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

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 other embodiments.

A further embodiment provides an analysis cartridge having pins to be connected 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.

Another embodiment provides a method for controlling an analysis system using the common heating layer.

side aspect

Embodiments belonging to a side aspect provide an analysis system for analyzing an analyte. The analysis system comprises a chamber and a heater, 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 heater is configured to deliver 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.

Embodiments according to this aspect are based on the finding that a heating device having a first geometry radiates heat in the direction of the chamber in a different geometry due to the thermal conductivity on 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 the gradients in the chamber.

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.

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 enables 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, enables a high gradient from the center to the edge. The gradient of the second case is in the middle between the first and the second case.

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, such that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid in the chamber. Based on this, not only the circulation of the fluid can be achieved, but also degassing of an adhesive material in the layer of the chamber can be avoided or reduced.

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, the heater 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 configured such that is chosen so that a temperature gradient from the center to the edge region of the chamber causes a circulation of a fluid in the chamber.

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 that a rapid cyclic temperature change is achieved without increasing the complexity of the entire 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 is arranged as a layer on the main surface of the chamber.

side aspect

An embodiment of this aspect of the invention provides an analysis system for analyzing an analyte. The analysis system comprises at least one chamber, a semi-transparent 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 indicative of an optical activity of the analyte.

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 has high functionality, high detection accuracy and low manufacturing costs. This concept has additional advantages in terms of usability, since the analysis cartridge does not have to be positioned or aligned with respect to an external optical detector.

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 manufacture of the detector and the light source directly on the microfluidic system, it is possible to increase the 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 that is configured to convert a light source into a background with homogeneously distributed illumination. Here, a frequency change can also be carried out such that a light frequency matches an absorption spectrum of the optical detector.

According to a further embodiment, the analysis system can be coupled to a processor or have a processor that enables the light transmission behavior and/or the optical activity of the analyte to be analyzed. A light transmission through the chamber that has a low absorption indicates the case in which no reaction has taken place, a light transmission through the chamber that has a high absorption indicates the case in which a reaction has taken place. Fluorescence, chemiluminescence or colorimetry of the analyte indicates the case in which a biochemical reaction is in progress.

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.

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

Another embodiment provides an analysis cartridge comprising an analysis system and pins 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 possible.

A further embodiment provides a graphically displayable diagnostic system comprising an analysis system according to one of the above embodiments and a battery as a power supply. Here, a small battery is sufficient, since the heating devices for heating the chambers and for controlling the elements are arranged directly next to the chamber or the temperature-controllable elements, so that the power consumption thereof is quite low.

• 1 ein Blockdiagramm einer Grundimplementierung eines Analysesystems eines Nebenaspekts zeigt;
• 2a-2f Diagramme zum Beschreiben des Hintergrundes der Ausführungsbeispiele gemäß dem Aspekt der 1 zeigen;
• 3 ein Blockdiagramm einer Grundimplementierung eines Analysesystems gemäß dem Hauptaspekt zeigt;
• 4 ein Blockdiagramm einer verbesserten Implementierung des Analysesystems gemäß dem Aspekt der 3 zeigt;
• 5 ein Blockdiagramm einer Grundimplementierung eines Analysesystems gemäß einem weiteren Nebenaspekt zeigt;
• 6 ein Blockdiagramm einer verbesserten Implementierung des Analysesystems gemäß dem Aspekt der 5 zeigt;
• 7a-7c drei verschiedene Blockdiagramme zum Veranschaulichen des optischen Transmissionsverhaltens in der Kammer für drei unterschiedliche Fälle gemäß einem Aspekt der 5 zeigen;
• 8a-8g eine weitere verbesserte Implementierung eines Analysesystems gemäß einem Aspekt der 5 veranschaulichen; und
• 9a, 9b auf eine herkömmliche Implementierung eines Analysesystems Bezug nehmen.

Embodiments of the present invention will now be explained with reference to the accompanying drawings, in which:

• 1 shows a block diagram of a basic implementation of a side aspect analysis system;
• 2a-2f Diagrams for describing the background of the embodiments according to the aspect of 1 show;
• 3 shows a block diagram of a basic implementation of an analysis system according to the main aspect;
• 4 a block diagram of an improved implementation of the analysis system according to the aspect of 3 shows;
• 5 shows a block diagram of a basic implementation of an analysis system according to a further side aspect;
• 6 a block diagram of an improved implementation of the analysis system according to the aspect of 5 shows;
• 7a-7c three different block diagrams to illustrate the optical transmission behavior in the chamber for three different cases according to an aspect of the 5 show;
• 8a-8g a further improved implementation of an analysis system according to an aspect of 5 illustrate; and
• 9a , 9b refer to a conventional implementation of an analysis system.

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

1 View A shows a cross-sectional view of an analysis system 10 for analyzing an analyte 13 according to a side aspect and a view B shows a top view of the analysis system 10. The analysis system 10 has a chamber 14, which can be embedded in a layer, which comprises, for example, polymer films. This optional layer is designated by the reference numeral 14l. In addition, the analysis system 10 has a heating device 12, which is integrated in a layer 12l, which is located next to the chamber 14 or next to the layer 14l of the chamber 14.

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

As with regard to 1b As can be seen, the heater 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 releases heat to the chamber 14 and its layer 14l. Therefore, a temperature distribution characteristic of the device 12 extends in the direction towards the chamber 14.

As a result of this two-dimensional geometry of the heating device 12, the shape of the temperature distribution characteristic designated by the 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 middle and is reduced 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 rather defined as a smooth transition. However, the transition bandwidth within this transition region decreases above average, so that the shape of the temperature distribution characteristic 12tc can be clearly defined.

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 constants of the temperature 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.

Regarding the size of the chamber 14, it should be noted that the size in the two-dimensional region is essentially related to the size of the two-dimensional temperature distribution characteristics ristik 12tc. Here, this essentially means that the chamber can be smaller, e.g. 30% smaller, the same size as, 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.

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.

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) may be used.

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 2a The high PCR efficiency is shown in 2b illustrated.

2b shows a graph of post-qPCR analysis results for four chips amplified using a slide-based µPCR system.

Since the three different steps in the PCR cycle (melting, extension and denaturation) are each initiated by a specific temperature, it is essential to expose the sample to as homogeneous and precise a temperature as possible throughout the experiment. In the PCR system, the hot spot of a (grid copper) heater is used to define the shape of the PCR chamber. By varying the size of the microPCR chamber, chambers with either constant temperature or different constant gradients can be achieved.

2c shows a hot spot 12tc' in a 4 × 6 mm grid heater 12'. The hot spot 12tc' generated in a 4 × 6 mm grid heater with a 30 µm line and a 300 µm space, as monitored with thermochromic liquid crystals (TLC), has a bandwidth in the range of 60° to 65° C. Here the hot spot 12tc' is oval.

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 2d is illustrated.

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 each other in terms of their size. All chambers 14'a, 14'b and 14'c are oval, 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 having a width of 4.8 mm, i.e. larger than the heater 12'.

For each design A, B and C, the resulting temperature gradient from the center to the edges is given by 2e where each chamber is filled with TLC having a temperature range of 60° to 65° C.

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, here 1° to 1.5° C from the center to the edge, whereas the gradient of design C is quite large, i.e. 3° to 4° C from the center to the edge. Design B has a gradient 2° to 3° C from the center to the edge.

Exposure to a homogeneous or slight gradient may have a beneficial effect depending on the type of PCR experiment. A homogeneous temperature may prevent nonspecific primer binding in some PCR experiments, whereas a gradient of several degrees may induce flow in the sample fluid between cold and warm regions and may have a stirring effect that promotes exchange between the chemical components in the fluid.

Challenges in microPCR include achieving precise thermal control and rapid thermal cycling in a polymer with low thermal conductivity. Similar results have been achieved recently 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. In this way, 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 ( 2f) .

Furthermore, the heater used only requires a 2 V supply with a maximum current setting level of 150 mA, which means that it could be powered by a small battery. Through this µPCR chip design, we enable a simple structure suitable for integration into portable molecular diagnostic systems.

With reference to 3 the main aspect is explained in detail. 3 shows an analysis system 10' comprising the chamber 14 and a further element 15, e.g. a valve, arranged next 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, which is 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 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.

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 by means of which the processing in the chamber 14 is supported or controlled, the layer 12a can be referred to as a control layer. From another point of view, this means that almost 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, so that the complexity, especially the complexity in terms of manufacturing effort, is reduced.

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.

According to further embodiments, the analysis system 10' may comprise a plurality of chambers, e.g., different heaters would also be arranged in the layer 12l for different purposes. This embodiment, which enables a fully integrated microfluidic sample-to-response system for nucleic acid testing, requires 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 multiple components of whole blood (filtration/cell sorting mechanism), breaking the cell membrane to extract DNA (cell lysis), amplifying the DNA, and detecting. This requires multiple functionalities such as dissolving and mixing reagents, filtering solutions, and incubating samples. Handling the processes in sequential order generally also requires valve mechanisms and waste disposal. Multiple approaches to enabling each assay 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, integration of functionalities comes at the cost of increased manufacturing costs.

A schematic principle of such an analysis, which has several sequential steps, is given by 4a The implementation in a 10'' laboratory using a microfluidic section is shown by 4b and 4c illustrated. 4b shows a side view, where 4c shows a top view of the analysis system 10''. The analysis system 10'' includes a plurality of reaction chambers 14, 14''b, 14''d and 14''f connected by channels or, more specifically, by controllable channels that are controllable using temperature controllable elements discussed below. To control the temperature-controllable elements and to heat the chambers 14 to 14''f, a control layer 12''l, which has a plurality of heating devices 12''a to 12''f, is integrated into the analysis system 10''.

The elements 12''a to 12''f are thin film copper heaters which are compatible with the heaters described above with respect to the aspect of 1 discussed above. Element 12''b in combination with chamber 14''b enables cell dissolution, element 12''d in combination with chamber 14''d enables DNA amplification using PCR, 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 to 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 have the purpose of controlling 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 comprising a wax plug, e.g. an irreversible wax plug, which can be melted using the heaters 12''b to 12''e. Alternatively, the heat-induced valves 15''a to 15''e can comprise expansion/compression elements made, for example, from heat-sensitive polymers.

The advantage of the thermal control-only approach is that most of the functionality can be created in a single process step, such as a single layer of copper, which greatly reduces the complexity of the manufacturing process. The solution could enable a truly low-cost probe-to-response diagnostic system.

With reference to 5 another aspect of the invention is discussed. 5 shows an analysis system 10''' comprising the chamber 14, e.g. in the 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, ie 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.

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

The optical detector 20 may 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.

According to further embodiments, the analysis system 10''' can have a light-emitting component 21, which is preferably arranged on the side of the heating device, i.e. opposite the optical detection layer 20l.

Whether the microfluidic system provides the platform for a cell cultivation experiment, detects a certain pathogen in a diagnostic test, or represents a microbioreactor for the production of a bioproduct, the result of the biochemical processes must be reported to the macro world qualitatively and/or quantitatively. Detection of analytes and produced products is generally performed 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 point-of-care setups, 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 that enable effective multiplexing.

With reference to 6 An improved embodiment is discussed. 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.

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 For this purpose, the optical layer 20l is implemented as a layer stack comprising a foil substrate 20f, an electro-optically sensitive layer 20, integrated electrodes 20i and an encapsulation material 20e. Regarding the layer 14l, note that the chamber 14 is formed in an adhesive foil 14f. From another point of view, this means that the semi-transparent thin film microheater is fabricated on a foil substrate 1. The optical detection layer is fabricated on a foil substrate 2. The two foils are arranged in a superimposed manner over a microfluidic cavity by a double-sided adhesive foil into which the contour of a microfluidic channel has been cut. The semi-transparent heater allows the light source and the 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 take place either in liquid form or in immobilized form on either substrate 1 or 2.

Regarding the 7a to 7c The three possible events that can be detected using the detector are discussed. Here 7a to 7c an analysis system 10'''', where 7a a light path from the light source 21 to a detector 20 without any biochemical activity in the chamber 14. 7b shows a light path of a biochemical activity having a blocking effect on light passing from the light source 21 to a detector 20. 7c shows a light path after a chemical activity that produces a change in the wavelength of the light from the light source 21 to a detector 20.

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 7b , causing a reduction in the light reaching the detector (absorbance measurement), or a change in the wavelength of the light arriving at the detector, as in 7c (fluorescence, chemiluminescence or colorimetric measurement). In the case of an experiment such as 7c Optionally, optical thin film filters can be produced by coating the substrate 1 ( 6 ), or the polymer material of the substrate 1 may be chosen to have optical filter characteristics to enable excitation of light of the correct wavelength for the chosen marker.

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

8a shows a further analysis system 10''''' or, in detail, an electro-optical measurement setup for a DNA hybridization detector that uses bead labeling. Here, the detector layer 20l' has an OSC layer (OSC = organic semiconductor layer) 20o, carbon fingers 20c and a (PET) film 20f.

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, which is arranged between the layer 14l having the chambers 14' and the LED 21l.

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

Here, the sensor layer 14l' also has a (PEN) film 14f, which is arranged on the spacer and encloses the cavity 14. The cavity 14 has the purpose of receiving the analyte 13 (here a DNA + beads) in such a way that it can be analyzed using the detector 20l'.

This detection is carried out with reference to 8b to 8g discussed in detail.

8b shows schematically the detector of the 8a in two different cases, namely in the case of a detected match and in the case of a mismatch during DNA detection or detection of hybridization events. Using biotin-labeled DNA targets and streptavitin-coated polymer beads of 1 µm size, it is possible to obtain different concentrations of a DNA targets. In the case of mismatch in the 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' passing through the foil 14f is not absorbed in the chamber 14, so that the emitted light reaches the detector 20 in an almost non-absorbed manner.

In case of a match, the streptavitin-coated batch 15 having 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, whereby a high absorption of the light has occurred.

8c shows the response to bead labeling based on four different cases where samples A1, A2 and A3 were hybridized to a 1 µm biotinylated target, whereas B1, B2 and B3 were hybridized to 50 nm and C1, C2 and C3 were hybridized to 2.5 nm. The two mismatch samples Mismatch1 & Mismatch2 with no beads bound 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.

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

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 designed 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 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 between a range that depends 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.

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

In addition, 8g the property of the optical film layer 14f to be able to convert a point light source such as an LED 21l into a homogeneously illuminated background that 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.

All 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 located right next to the chamber, embodiments according to all aspects have low power consumption so that they can be battery operated. Therefore, another embodiment provides a portable diagnostic system comprising one of the analysis systems discussed above and a battery as a power supply. Combining the elements into a microfluidic system would enable an analysis module for biological or chemical events that is extremely inexpensive and portable.

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 has, for example, 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.

Although the above embodiments have been discussed primarily in connection with a device, it should be noted that further embodiments provide corresponding methods. Therefore, according to a further Embodiment provides a method for controlling an analysis system.

This method begins in the embodiments discussed in relation to the main aspect and comprises the steps of heating at least one chamber to effect 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 a low absorption indicates an instance in which no reaction has occurred, wherein a signal corresponding to a light transmission through the chamber having a high absorption indicates an instance in which a reaction has occurred. Another signal corresponding to fluorescence, chemiluminescence or colorimetry or something else indicates an instance in which a chemical or biochemical reaction is in progress.

It is understood that aspects discussed in connection with a device also represent a description of the corresponding method, where 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 or 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.

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

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.

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 carry out one of the methods when the computer program product runs on a computer. The program code can, for example, be stored on a machine-readable carrier.

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

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.

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 carrying out one of the methods described herein is recorded. The data carrier, the digital storage medium or the recorded medium are usually tangible or non-volatile.

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

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.

A further embodiment comprises a computer on which the computer program installed to perform any of the methods described herein.

A further embodiment according to the invention comprises a device or a system which is designed to transmit a computer program for carrying out 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.

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 cooperate with a microprocessor to perform any of the methods described herein. In general, the methods are preferably performed by any hardware device.

The embodiments described above 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. [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. [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, 28 October - 1 November 2012, Okinawa, Japan, pages 797-799 .
3. [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. [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 (9)

An analysis system (10, 10', 10'', 10''', 10'''', 10''''') for analyzing an analyte (13), comprising: at least one chamber (14) for processing the analyte (13), the chamber (14) being formed by one or more polymer films and comprising 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. 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). An analysis system (10, 10', 10'', 10''', 10'''', 10''''') according to one of the preceding claims, wherein the heating device (12, 12a) differs in size from the further heating device (12c). An analysis system (10, 10', 10'', 10''', 10'''', 10'''') according to one of the preceding claims, wherein the at least one chamber (14) is designed for cell dissolution, DNA amplification, rapid DNA hybridization and target detection up to the signal base layer, cell isolation, DNA extraction tion, DNA detection and/or polymerase chain reaction. 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 has 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'). 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 adjacent to the at least one chamber (14) for detecting light coming from the chamber (14) and configured to output a signal indicating a transmission behavior through the chamber (14) or indicating an optical activity of the analyte (13). Analysis cartridge comprising an analysis system (10, 10', 10'', 10''', 10'''', 10'''') according to 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. 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. 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), the chamber (14) being formed by one or more polymer films and comprising 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) which has 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) in order to carry out the processing of the analyte (13); and Heating the temperature-controllable element (15, 15``a, 15``c, 15``e) in order to control the same.

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