DE102016211355A1 - Analysis system and method for performing an analysis - Google Patents

Abstract

Embodiments according to the main aspect are based on the finding that a heater having a first geometry radiates heat in the direction of the chamber in a different geometry due to the thermal conductivity at the edges of the heater. Based on this knowledge, the shape of the chamber is adapted to the shape of the temperature distribution characteristic and not to the shape of the heater. Such a concept has advantageous effects on the temperature behavior and especially on the constants and the gradients in the chamber.

Description

One aspect of the invention relates to an analysis system having a heater. Preferred embodiments relate to an analysis cartridge or portable diagnostic systems having an analysis system.

Lab-on-a-chip (Lab-on-a-Foil), micro-total-analysis-systems (μTAS), or bio-micro-electro-mechanical (Bio-MEMS) systems are common names for Such systems, which deal with the miniaturization of chemical and biological laboratory processes and experiments in micro scale format, with the aim to improve the efficiency of the experiment and to reduce its cost and time. Common to these systems is that they handle liquids since 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 final products to be able to analogously model a corresponding range of laboratory equipment. The various functionalities may be provided to the microfluidic system either by miniaturization and integration as on-chip components, or may be kept separate as off-chip macroscale components, with a macro-to-micro interface that integrates functionality into the microfluidic device having advantages such as Improved performance and sensitivity (direct contact with the sample) and reduction of bulky off-chip components (important for portable applications, PoC), but at an increased cost, because on-chip integration involves additional processing steps.

To avoid contamination risks, microfluidic systems are often disposable disposable systems requiring low material and manufacturing costs, and thus plastics and polymers are preferred over the classic MEMS silicone material. Micro injection molding is the current state-of-the-art manufacturing technology for mass production of these systems. Despite the ability to produce millions of high throughput microfluidic plastic substrates, the process has difficulty integrating functionalities. The injection molded microfluidic system is generally made by solvent or thermal bonding, which is often incompatible with electrodes and biomolecules to be integrated. Use of plastic films and roll-to-roll production is an attractive alternative mass production process because integration of such elements through the use of lamination-based processes and pressure-sensitive adhesive tapes is readily accomplished with high throughput. This allows various materials to be bonded together in a mix-and-match approach, and any component of the system can be manufactured under 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 culture experiment require a constant 37 ° C. In an experiment of DNA hybridization, the target and analyte strand hybridize only under incubation 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 60-65 ° C. Rolling circle amplification (RCA) requires a denaturation step at 95 ° C before extension at 30 ° C, and finally, the polymerase chain reaction (PCR) uses cyclic events between ~ 63 ° C, ~ 72 ° C and ~ 95 ° C for exponential amplification of the template DNA. In order to obtain precise temperature control and high heating and cooling rates, integration of micro-scale heaters has improved performance compared to external macroscale heaters due to direct thermal contact with the microfluidics.

We have already 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]. It has been found that the microheater provides a robust and homogeneous temperature at the point of analysis, can be made roll-to-roll (low cost) and requires 2V power input (which is why it is portable).

A conventional chip lab is in 9a shown. 9a shows an analysis system 10 * (also called chip lab), which is a heating device 12 * , z. B. a copper heater on a foil and a chamber 14 * for processing analytes. The chamber may be, for example, a DNA microarray, such as by enlarging the chamber 14 * is shown. For transporting the analytes, here the DNA having analytes, into the chamber 14 * must be the layer of the chamber 14 * possibly one or more microfluidic channels 14mf * exhibit. The chamber 14 * is formed in a layer next to the the heater 12 * having layer. For example, layer of the chamber 14 * and the layer of the heater 12 * using a structured double-sided adhesive film 14a * be attached to each other. In addition, the layer of the chamber 14 * through a foil cover (made of PET / acrylic / PEN) 14c * be covered, which is arranged on one side of the chamber, that of the heating device 12 * facing layer.

The analysis system 10 * The purpose is to perform a detection of SNPs using a melting curve analysis. Melting curve analysis is an evaluation of a dissociation characteristic of a double-stranded DNA during heating. As the temperature increases, the duplex 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 graphs showing the dependence between fluorescence and a temperature at which fluorescence occurs. The melting curves shown belong to two groups, namely to a first group with 20m * is marked and matches, and to a second group with 20mm * is marked and shows mismatches. To such melting curves 20m * and 20mm * to detect the temperature of the analyte is increased fairly precisely, the analyte z. B. is monitored using an optical detector with respect to its fluorescence or with respect to another optical effect such as colorimetry or chemiluminescence.

As for the diagram of the 9b As can be seen, which illustrates the melting curve analysis, there is a need for fairly accurate temperature matching.

Solution approaches according to the prior art, for. B. the above-discussed solution approach of the analysis system 10 * , are often detrimental to the temperature constant in the chamber or to the gradients across the lateral dimensions of the chamber. Another common disadvantage is the optical detector and its ability to accurately monitor the optical effect. Furthermore, the analysis system often causes high manufacturing costs. Therefore, there is a need for an improved approach.

key aspect

An object of the present invention is to provide a concept for improving the temperature behavior in the chamber.

This object is solved by the subject matter of independent claims.

Embodiments pertaining to the main aspect provide an analysis system for analyzing an analyte. The analysis system comprises a chamber and a heating device, e.g. B. a (copper) grid heater, which is integrated into a layer and a two-dimensional geometry, for. As a square has. The layer is arranged next to the chamber. The heater is configured to have a shape according to a two-dimensional temperature distribution characteristic, e.g. As an oval shape, has to deliver heat to the chamber. 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. B. an oval shape.

Embodiments according to the main aspect are based on the finding that a heater having a first geometry radiates heat in the direction of the chamber in a different geometry due to the thermal conductivity at the edges of the heater. Based on this knowledge, the shape of the chamber is adapted to the shape of the temperature distribution characteristic and not to the shape of the heater. Such a concept has advantageous effects on the temperature behavior and especially on the constants and the gradients in the chamber.

According to embodiments, the aspect ratio of the two-dimensional Temperature distribution characteristic 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 may be comparable or substantially identical. Here a distinction can be made for different cases. According to a first case, the chamber is 10% to 30% smaller than the temperature distribution characteristic. This case allows a very small gradient from the center to the edge, resulting in an almost homogeneous distribution over the entire chamber. In a second case, the temperature distribution characteristic is within a tolerance of ± 20% identical to the chamber. A third case, according to which the chamber is 10% to 30% larger than the temperature distribution characteristic, allows a high gradient from the center to the edge. The gradient of the second case lies midway between the first and second cases.

In other embodiments, the chamber is configured for a polymerase chain reaction wherein the chamber is greater 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 fluid in the chamber. From this not only the circulation of the fluid can be achieved, but also a 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 includes a polymerase chain reaction chamber and a heater integrated into a chamber adjacent the chamber and having a two-dimensional geometry, wherein the heater is configured to apply heat to the body in accordance with a two-dimensional temperature distribution characteristic having a shape Chamber, 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 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 has 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 heater is implemented as a thin-film heater and disposed as a layer on the main surface of the chamber.

Second aspect

An embodiment according to another 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. In the control layer, a heating device for heating the at least one chamber and a further, coupled to the temperature-controllable element heater is implemented. The heating device for heating the at least one chamber has the purpose of assisting processing in the chamber or enabling processing, whereas the further heating device coupled to the temperature-controllable element is configured to supply the temperature-controllable element by heating it Taxes. According to preferred embodiments, the heating device and the heating device are formed by a common heating layer.

Embodiments of a further aspect are based on the recognition that almost all or all elements can be implemented as temperature-controllable elements, so that no further elements for controlling the chip laboratory have to be implemented. This is advantageous in terms of the complexity of the analysis system.

According to embodiments, at least one chamber may be configured for cell dissolution, DNA amplification, rapid DNA hybridization and target detection to a signal base layer and / or polymerase chain reaction. The temperature-controllable element may for example be a temperature-controllable valve, z. 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 usually have different operating temperatures, the size of the heaters in the common layer may vary depending on further embodiments.

Another embodiment provides an analysis cartridge having pens to be connected to a reading device that controls and / or evaluates the analysis and an analysis system. Here, the pins are connected to the heater and the further heater of the control layer so that the analysis cartridge can be externally controlled.

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

Third aspect

An embodiment of this ancillary 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 in a layer adjacent to the chamber for detecting light from the chamber. The optical detector is configured to output a signal indicative of transmission behavior (permeability behavior) through the chamber or indicative of an optical activity of the analyte.

Embodiments according to another aspect are based on the recognition that the integration of a chamber and a semi-transparent heater on two different sides of the chamber enable an analysis system that has high functionality, high detection accuracy, and low manufacturing cost. This concept also has usability advantages because the analysis cartridge need not be positioned or aligned with respect to an external optical detector.

According to another embodiment, a light source on one side of the heater, i. H. be arranged 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 another embodiment, the light source may be used in combination with an optical filter configured to convert a light source to a background with homogeneously distributed illumination. Here also a frequency change can be carried out such that a light frequency coincides with 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 makes it possible to analyze the light transmission behavior and / or the optical activity of the analyte. A light transmittance through the chamber having a low absorption indicates the case where no reaction has occurred, a light transmittance through the chamber having a high absorption indicates the case where a reaction has occurred. Fluorescence, chemiluminescence or colorimetry of the analyte indicates the case of an on-going biochemical reaction.

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

All the above embodiments according to the above aspects relate to an analysis system using heaters. From this it is clear that the technologies described above are incompatible in aspects, but must also be combined in a predetermined way. Therefore, further embodiments provide an embodiment having features of at least two aspects.

Another embodiment provides an analysis cartridge having an analysis system and pens to be connected to a reading device that controls and evaluates the analysis. Here, the pins are connected to the optical detector so as to allow external access to the detector.

Another embodiment provides a graphically displayable diagnostic system that includes an analysis system according to any of the above embodiments and a battery for powering. Here, a small battery suffices since the heaters are arranged to heat the chambers and to control the elements directly adjacent to the chamber or temperature-controllable elements, so that the power consumption thereof is quite low.

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

1 shows a block diagram of a basic implementation of an analysis system of the main aspect;

2a - 2f Diagrams for describing the background of the embodiments according to the aspect of 1 demonstrate;

3 shows a block diagram of a basic implementation of an analysis system according to another subsidiary aspect;

4 a block diagram of an improved implementation of the analysis system according to the secondary aspect of 3 shows;

5 shows a block diagram of a basic implementation of an analysis system according to another subsidiary 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 illustrating the optical transmission behavior in the chamber for three different cases according to the aspect of 5 demonstrate;

8a - 8g a further improved implementation of an analysis system according to the aspect of 5 illustrate; and

9a . 9b refer to a conventional implementation of an analysis system.

Hereinafter, embodiments of the present invention will 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 the main aspect and shows in a view B a plan view of the analysis system 10 , The analysis system 10 has a chamber 14 on, in a layer z. B. polymer films may be embedded. This optional layer is denoted by the reference numeral 14l designated. In addition, the analysis system points 10 a heater 12 on that in a layer 12l integrated, located next to the chamber 14 or next to the shift 14l the chamber 14 located.

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

As for 1b can be seen, the heater has 12 , z. B. a copper grid heater, a two-dimensional geometry, z. As a square, on. The square of the grid heater 12 is parallel to the layer 12l ie parallel to the layer 14l arranged. 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 emits or gives heat to the chamber 14 and her shift 14l from. Therefore, a temperature distribution characteristic of the device extends 12 in the direction of the chamber 14 ,

As a result of this two-dimensional geometry of the heater 12 is also the shape of the reference number 12TC designated temperature distribution characteristic two-dimensional. However, the shape of the two-dimensional temperature distribution characteristic is different 12TC from the two-dimensional geometry of the heater 12 , Here, the two-dimensional temperature distribution characteristic 12TC an oval shape. The background for this round shape 12TC is that the heat dissipation concentrates in the middle and is reduced towards the boundary areas. In this context, it should be noted that the limit or shape of the temperature distribution characteristic 12TC has no sharp transition; it is defined as a smooth transition. However, the transition bandwidth within this transition region decreases more than average, so that the shape of the temperature distribution characteristic 12TC can be clearly defined.

On the basis of this knowledge, the shape and size of the chamber 14 to the two-dimensional temperature distribution characteristic 12TC be adjusted to 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 to achieve. In this embodiment, the chamber 14 an oval shape, that is, a shape having the shape of the two-dimensional temperature distribution characteristic 12TC is comparable. Alternatively, the shape of the chamber 14 hexagonal or can be compared to the shape of the temperature distribution characteristic 12TC have any other shape. Note that the above description regarding the shape of the chamber 14 on the two-dimensional shape in the region of the layer 14l Reference, ie 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 area substantially coincides with the size of the two-dimensional temperature distribution characteristic 12TC matches. Here essentially means that the chamber is smaller, z. B. 30% smaller, the same size as or larger, z. B. 30% larger than the temperature distribution characteristic 12TC can be. The exact dependence between the two sizes 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 with the projection of the chamber 14 to the plane 14l be compared. Alternatively, the mismatch between the boundaries can be compared. To 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 are similar 14 and 12TC , wherein the aspect ratio differs by a maximum of a factor of 1.5. Another method is to align (x, y) the boundaries of the two elements 14 and 12TC to compare.

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

According to further embodiments, the analysis system 10 a film-based microfluidic PCR module to which a thin film (copper grid) device for MikroPCR 12 was created. And PCR was like in 2a shown performed. The high PCR efficiency is in 2 B illustrated.

2 B shows a diagram of results of a post-qPCR analysis for four chips, which are amplified using a film-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 throughout the experiment to the most homogeneous and accurate temperature possible. 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 MikroPCR chamber, chambers of either constant temperature or different constant gradients can be achieved.

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

Starting from this heater 12 ' allow three different sizes of PCR chamber 14'a . 14'b and 14'c from the middle to the edge different temperature gradients, such as through 2d is illustrated.

2d illustrates three designs (draft A, draft B and draft C), the chambers 14'a . 14'b and 14'c correspond. The three designs A, B and C differ in size from each other. 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, ie 0.8 mm less than the width of the heater 12 ' , The design B has a width of the chamber 14'b of 4 mm, ie equal to the width of the heater 12 ' , The draft C represents a chamber 14'c ready, which has a width of 4.8 mm, ie larger than the heater 12 ' ,

For each design A, B and C, the resulting temperature gradient is from the center to the edges 2e shown, each chamber is filled with TLC, which have 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 is big, d. H. 3 ° to 4 ° C from the middle to the edge. The design B has a gradient of 2 ° to 3 ° C from the center to the edge.

Contacting with a homogeneous or light gradient may have a beneficial effect, depending on the nature of the PCR experiment. A homogeneous temperature may prevent nonspecific primer binding in some PCR assays, whereas a multiple grade gradient may cause flow in the sample fluid between cold and warm regions and may have a stirring effect that promotes an exchange between the chemical components in the fluid.

Challenges at MikroPCR are achieving precise thermal control and rapid cyclic temperature cycling for a low thermal conductivity polymer. Similar results have recently been achieved with polymer-based systems, which, however, require rather complex constructions, including liquid nitrogen reservoirs and air hoses, resulting in extensive instrumentation and increased costs. Use of 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 a low ambient temperature. In this way, the film-based PCR chamber allows a high ramping and cooling rate without the need for additional external forces. For example, using a simple convection device, a cooling rate of ~ 4 ° C / sec. (ie 95 to 63 ° C in ~ 8 seconds) ( 2f ).

Further, the heater used requires only a 2V supply with a maximum current setpoint level of 150mA, meaning that it could be powered by a small battery. With this μPCR chip design, we enable a simple design suitable for integration into portable molecular diagnostics systems.

With reference to 3 another aspect is explained in detail. 3 shows an analysis system 10 ' that the chamber 14 and another element 15 , z. B. a valve which is arranged adjacent to the chamber has. The element 15 is a temperature controllable element. Both elements 14 and 15 are in the layer 14l arranged. In a second shift 12l on the shift 14l is arranged, is a plurality of heating devices, namely the heater 12 and the heater 12c arranged. The heater 12 belongs to the chamber 14 , where the design of the heater, or more precisely, the design of the chamber 14 , can be implemented as described above with reference to 1 was discussed. The heater 12c belongs to the temperature-controllable element 15 ie the heater 12c is so in the layer 12l arranged that the heater 12c and the temperature-controllable element 15 aligned with each other.

The temperature-controllable element 15 , z. As a valve with a wax plug can, by using the heater 12c generated heat to be controlled. As the heater 12c in the same layer 12l is arranged, by means of which the processing in the chamber 14 supported or controlled, the layer can 12a be referred to as tax layer. Viewed from a different angle, this means that almost every element of the analysis system 10 ' controlled by using heaters. Thus, it is advantageous that the heaters are formed by the same metal layer, so that the complexity, in particular the complexity in terms of manufacturing cost, is reduced.

Because the chamber 14 usually requires a different temperature than the temperature-controllable element 15 , the heaters can 12 and 12c be different in terms of their size, or generally with respect to their heating power.

According to further embodiments, the analysis system 10 ' have a plurality of chambers, for. For example, different heaters would be in the layer for different purposes 12l arranged. This embodiment, which enables a fully integrated microfluidic sample-to-response system for nucleic acid assays, requires manipulation of microvolumes of fluids in several consecutive steps. For example, detection of a specific pathogen by its DNA in a patient's blood sample involves sorting out the pathogen from the various components of the whole blood (filtration / cell sorting mechanism), disrupting the cell membrane to extract DNA (cell dissolution), amplifying the DNA, and a detection. This requires a variety of functionalities, such as dissolving and mixing reagents, filtering solutions, and incubating samples. Handling the processes in sequential order also generally requires valve mechanisms and waste disposal. A variety of approaches to enabling each trial step are possible. For example, a cell membrane (lysis) can be disrupted by exposure 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 has the cost of increased manufacturing costs.

A schematic principle of such an analysis, which has several sequential steps, is through 4a illustrated. The implementation in a laboratory 10 '' that uses a microfluidic cut is through 4b and 4c illustrated. 4b shows a side view, wherein 4c a plan view of the analysis system 10 '' shows. The analysis system 10 '' has a plurality of reaction chambers 14 . 14''b . 14''d and 14''f which are connected by channels or, more specifically, by controllable channels which are controllable using temperature-controllable elements which will be discussed below. To control the temperature-controllable elements and to heat the chambers 14 to 14''f is a tax layer 12''l containing a plurality of heaters 12''a to 12''f has, in the analysis system 10 '' integrated.

The Elements 12''a to 12''f are thin film copper heaters comparable to the heaters discussed above with respect to the main aspect. The element 12''b in combination with the chamber 14''b allows a cell resolution, the element 12''d in combination with the chamber 14''d a DNA amplification by PCR, the element 12''f in combination with the chamber 14''f allows fast targeting Detection using DNA hybridization and / or melting curve analysis for SNP detection. The Elements 12''a . 12''c and 12''e provide heat to the heat-induced valve mechanisms 15''a . 15''c and 15''e to control. The Elements 15''a With 15''c and 15''e between the chambers 14 and 14''b . 14''b and 14''d as well as between 14''d and 14''f are arranged, have the purpose of lateral flow between the different reaction chambers 14 . 14''b . 14''d and 14''f to control. Heat-induced valve 15''a to 15''e could be implemented as valves containing a wax plug, e.g. As an irreversible wax plug, comprising, using the heaters 12''b to 12''e can be melted. Alternatively, the heat-induced valves 15''a to 15''e Have expansion / compression elements, which are made for example on the basis of heat-sensitive polymers.

The advantage of the heat-only control approach is that most of the functionality can be generated in a single process step, such as a single layer of copper, which greatly reduces the complexity of the manufacturing process. The solution could allow for a truly cost-effective sample-to-response diagnostic system.

With reference to 5 another aspect of the invention is discussed. 5 shows an analysis system 10 ''' that the chamber 14 , z. In the layer 14l , wherein the layer 12l the one or more heaters for the one or more chambers 14 having. In addition, the analysis system points 10 ''' an optical detector layer 20l on, which is an optical detector 20 which is arranged and configured to from the chamber 14 to detect incoming light. According to a preferred embodiment, the optical detection layer 20l opposite the heating layer 12l arranged, ie such that the layer 14l containing the one or more chambers 14 between the two layers 20l and 12l is arranged. The chamber 14 and the heating layer 12l may be implemented according to the discussion of the main aspect and a minor aspect.

The optical detector 20 may be implemented according to the discussion of the main aspect and a minor aspect.

The optical detector 20 may be a simple optical detector having light dependent resistors or a CCD element. The detector 20 has the purpose of detecting whether a chemical reaction has taken place, is in progress or has not yet taken place.

According to further embodiments, the analysis system 10 ''' a light emitting device 21 preferably on the side of the heating device, ie opposite the optical detection layer 20l , is arranged.

Whether the microfluidic system provides the platform for a cell culture experiment, detects a certain pathogen in a diagnostic text, or represents a microbioreactor for the production of a bioproduct - the result of the biochemical processes must be qualitatively and / or quantitatively reported to the macro-world. Detection of analytes and generated products is generally by mass-sensing, electrochemical or optical means, making optical detection methods such as fluorescence, chemiluminescence, colorimetry or absorbance by far the most common. Integration of on-chip light sources and a photodetector for analyte detection in microfluidic systems, in addition to reducing the requirement of bulky optical components in near-patient structures, has the following advantages: 1) to make the detector and light source directly on the microfluidic system allow 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 patterned into condensed arrays that allow effective multiplexing.

With reference to 6 An improved embodiment will be discussed. 6 shows an analysis system 10 '''' containing the optical detector layer 20l , the heat generation layer 12l and the layer 14l comprising the microfluidic cavity.

In this embodiment, the heating layer is 12a implemented as a layer stack, which is a film substrate 12f , a thin film heater 12 and an encapsulating material 12e having. Analogous to this is the optical layer 20l implemented as a layer stack, which is a film substrate 20f , an electro-optically sensitive layer 20 , integrated electrodes 20i and an encapsulating material 20e having. Regarding the layer 14l it should be noted that the chamber 14 in an adhesive film 14f is formed. From another point of view, this means that the semitransparent thin-film microheater on a film substrate 1 is made. The optical detection layer is on a film substrate 2 produced. The two films are stacked over a microfluidic cavity through a double-sided adhesive sheet 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 houses the biochemical event. The biochemical process can be either in liquid form or in immobilized form either on the substrate 1 or 2 occur.

Regarding the 7a to 7c The three possible events that can be detected using the detector are discussed. Show here 7a to 7c an analysis system 10 '''' , in which 7a a light path from the light source 21 to a detector 20 without any biochemical activity in the chamber 14 shows. 7b shows a light path of biochemical activity with a blocking effect on light coming from the light source 21 to a detector 20 arrives. 7c shows a light path after a chemical activity, which is a change in the wavelength of the light source 21 to a detector 20 reaching light generated.

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 taken place in the microfluidic cavity, two detection mechanisms can be used, depending on the experimental setup. Either the biochemical event produces a product that produces steric hindrance to the light that comes from the light source and arrives at the detector, as in FIG 7b , wherein a reduction of the light reaching the detector is effected (absorbance measurement), or it will be a change in the wavelength of the incident light at the detector, as at 7c (Fluorescence, chemiluminescence or colorimetric measurement). In the case of a trial as in 7c Optionally, thin-film optical filters can be created by placing the substrate 1 is coated ( 6 ), or the polymeric material of the substrate 1 can be chosen to have optical filter characteristics to allow excitation of light of the correct wavelength for the selected marker.

The biochemical event could be that cell culture proliferates laterally and light from the light source is blocked, could be a DNA hybridization assay, an immunoassay or an amplification product of any of the DNA amplification protocols previously described. The hybridization assay or amplification product could be labeled with either fluorescence, chemoluminescence or colorimetric or fluorescence (polymer bead, gold nanoparticle, silver enhancement) labels.

8a shows another analysis system 10 ''''' or in detail an electro-optical measuring setup for a DNA hybridization detector using a bead labeling. Here is the detector layer 20l ' an OSC layer (OSC = organic semiconductor layer) 20o , Carbon finger 20c and a (PET) film 20f on.

Opposite the detector 20l ' is the light source layer 21 ' arranged. Here is the light source 21 through an LED 21l that z. B. is configured to output light in the range of 360 nm, and by an optical spacer 21s that between the chambers 14 ' having layer 14l and the LED 21l is arranged, formed.

Between the detector layer 20l ' and the light source layer 21 ' is the layer 14l ' arranged the chambers 14 (fluidic chamber or cavity 14 ) having. This layer 14l ' Can also be used as a sensor layer 14l ' be designated.

Here is the sensor layer 14l ' also a (PEN) foil 14f on, which is arranged on the spacer and the cavity 14 encloses. The cavity 14 has the purpose of the analyte 13 (here, a DNA + bead) to be taken up by using the detector 20l ' can be analyzed.

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

8b schematically shows the detector of 8a in two different cases, namely in the case of a detected match and in case of a mismatch during DNA detection or the detection of hybridization events. When using biotin-labeled DNA targets and streptavitin-coated polymer beads of 1 micron size, it is possible to detect various concentrations of a DNA target. In the case of disagreement in the chamber 14 No DNA and no target object were aligned with the DNA strand. As a result, it will be out of the light source 21 ' emitted light, which is the film 14f does not happen in the chamber 14 absorbed so that the emitted light is the detector 20 reached almost in a non-absorbed way.

In the case of correspondence, the streptavitin-coated batch reacts 15 having the complementary DNA strand with the DNA strand. This reaction can be detected due to the fact that the light source 21 ' emitted light the detector 20 achieved, wherein a high absorption of light has occurred.

8c Figure 4 shows the response to bead labeling due to four different cases in which samples A1, A2 and A3 were hybridized with a biotinylated 1 μm target, whereas B1, B2 and B3 were 50 nm and C1, C2 and C3 were 2.5 were hybridized. The two mismatch samples mismatch1 & mismatch2 without bound beads are used as control points. As you can see, the different concentrations cause different optically detectable effects, so that the detector 20 can distinguish between the concentrations.

An exemplary signal that belongs to three different concentrations and one mismatch is 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 , z. FS102 with an absorption maximum at 400 nm, and an LED light source of ~ 360 nm, 21l , An optical fiber is formed in the structure, the transmission and autofluorescence properties of the film layers 20f and 14f to use. The transmission spectra of the layers 20f and 14f are in 8e shown. 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 using an optical source 21l such that it lies between this region or generally between a region which depends on the selected film layer, 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 is consistent with the absorption spectra of the OSC layer 20o (~ 400 nm) matches. Whereas the film layer (PET) 20f permeability of the same without any change in the light.

8f Fig. 12 shows the spectra showing the autofluorescence property of the optical layer 20f and 14f due to the light source 21l specify.

Additionally shows 8g the property of the optical film layer 14f to be able to use a point light source such as B. an LED 21l to transform it into a homogeneously illuminated background for the one in the chamber 14 is more suitable on the basis of beads implemented light blocking. Whereas the film layer 20f on the chamber 14 allows the light to pass unhindered to the OSC layer 20o passes through.

All of the systems discussed above, which belong to different aspects, are preferably made entirely on film, for example using a roll-to-roll technology. Since the detector layer and the heating layer are arranged directly next to the chamber, embodiments in all aspects have a low power consumption, so that they can be battery operated. Therefore, another embodiment provides a portable diagnostic system having one of the analysis systems discussed above and a battery as a power supply. The combination of elements into a microfluidic system would allow a biological or chemical event analysis module that is extremely cost effective and portable.

Another embodiment provides an analysis cartridge having an analysis system according to one or more of the aspects discussed above. For example, the analysis cartridge system is in the form of a credit card and has outside pegs through which the analysis system can be connected to a reader to control / analyze the analysis. For example, the pins are inside with the control layer, i. H. connected to the heaters, or to the optical detector elements. The analysis cartridge may be a disposable element in which the analysis electronics could be separated, i. H. 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 located outside.

Although the above embodiments have been discussed primarily in the context of a device, it should be understood that other embodiments provide such methods. Therefore, according to another embodiment, a method for controlling an analysis system is provided. This method begins with the embodiments discussed in a minor aspect and includes the steps of heating at least one chamber to perform the processing of the analyte and heating the temperature-controllable element to control the same. Another method relates to analyzing an analyte. This method starts in the embodiments according to a side aspect and includes 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 a case in which none Reaction has taken place, wherein a signal that belongs to a light transmission through the chamber, which has a high absorption, indicates the case in which a reaction has taken place. Another signal leading to a fluorescence, Chemiluminescence or colorimetry or otherwise, is the case of a chemical or biochemical reaction in progress.

It is understood that the aspects discussed in connection with a device also represent a description of the corresponding method, wherein a block or a component corresponds to a method step or a feature of a method step. Similarly, aspects described in connection with a method step also represent a description of the corresponding block or feature 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 electronic circuit. In some embodiments, some or more of the most important 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 are stored on the electronically readable control signals, which cooperate with a programmable computer system (or can cooperate), that the respective method is performed. Therefore, the digital storage medium can be computer readable.

Some embodiments according to the invention include a data carrier having electronically readable control signals capable of interacting with a programmable computer system to perform one of the methods described herein.

In general, embodiments of the present invention may be implemented as a computer program product having a program code, wherein the program code is operable to perform one of the methods when the computer program product runs on a computer. The program code may for example be stored on a machine-readable carrier.

Other embodiments include the computer program for performing any 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 performing one of the methods described herein when the computer program runs on a computer.

A further embodiment of the inventive method is thus a data carrier (or a digital storage medium or a computer-readable medium) on which the computer program is recorded for carrying out one of the methods described herein. The data medium, 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 represent the computer program for performing one of the methods described herein. The data stream or the sequence of signals may be configured, for example, to be transferred via a data communication connection, for example via the Internet.

Another embodiment includes a processing device, such as a computer or a programmable logic device, that is configured or adapted to perform one of the methods described herein.

Another embodiment includes a computer on which the computer program is installed to perform one of the methods described herein.

Another embodiment of the invention includes a device or system configured to transmit (eg, electronically or optically) a computer program for performing at least one of the methods described herein to a receiver. The receiver may be, for example, a computer, a mobile device, a storage device or the like. For example, the device or system may include a file server for transmitting the computer program to the receiver.

In some embodiments, a programmable logic device (eg, 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 one of the methods described herein. Generally, the methods are preferably performed by any hardware device.

The embodiments described above are merely illustrative of the principles of the present invention. It will be understood that modifications and variations of the arrangements and details described herein will be apparent to others of ordinary skill in the art. Therefore, it is intended that the invention be limited only by the scope of the appended 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 substrates"; 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 28-1. November 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] Submitted patents incorporating the grid heater concept:
• - PCT / EP2013 / 071952
• - DE 10 2014 221 734.2
• - DE 10 2015 202 353.2 ;

Claims (15)

An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) for analyzing an analyte ( 13 ), comprising: a chamber ( 14 ) for processing the analyte ( 13 ); and a heating device ( 12 . 12 ' . 12''a . 12''b . 12''c . 12''d . 12''e . 12''f ), which is placed in one of the chambers ( 14 ) integrated layer ( 12l ) and has a two-dimensional geometry, wherein the heating device is configured according to a two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ), which has a shape to deliver heat to the chamber, which is in a chamber adjacent to the chamber ( 14 ) integrated layer ( 12l ) and wherein a lateral shape or lateral size of the chamber ( 14 ) to the shape of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) is adjusted. The analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to claim 1, wherein an aspect ratio of the chamber ( 14 ) potentially having an aspect ratio of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) matches; and / or in which the lateral shape of the chamber ( 14 ) substantially with the shape of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) matches. The analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to claim 2, wherein the aspect ratio of the chamber ( 14 within a tolerance of ± 20% with the aspect ratio of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) matches. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, in which the lateral size of the chamber ( 14 ) substantially with the size of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) matches. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to claim 4, wherein the chamber ( 14 ) is up to 10% to 30% smaller than the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ); and / or where the lateral size of the chamber ( 14 within a tolerance of ± 20% equal to the size of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ); and / or where the chamber ( 14 ) Is 10% to 30% larger than the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ). An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to claim 4, where the chamber ( 14 ) is configured for a polymerase chain reaction and in which the chamber ( 14 ) Is 10% to 30% larger than the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) such that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid in the chamber at the edge region. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, in which the chamber ( 14 ) is configured for a polymerase chain reaction or a micro-polymerase chain reaction. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, in which the chamber ( 14 ) is formed by one or more polymer films and / or has a polymer with low thermal conductivity. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, in which the grid heating device ( 12 . 12 ' . 12''a . 12''b . 12''c . 12''d . 12''e . 12''f ) is implemented as a thin film grid heater and is mounted on a major surface of the chamber (FIG. 14 ) is arranged. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, wherein the analysis system further comprises a temperature controllable element ( 15 . 15''a . 15''c . 15''e ) and a tax layer ( 12l ), in which the grid heating device ( 12 . 12 ' . 12''a . 12''b . 12''c . 12''d . 12''e . 12''f ) for heating the chamber ( 14 ), to perform the processing, and wherein another heater is coupled to the temperature-controllable element and configured to control the temperature-controllable element. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to any one of the preceding claims, further comprising an optical detector ( 20 ) located in one of the chambers ( 14 ) is integrated to remove from the chamber ( 14 ) and is configured to output a signal indicative of transmission behavior through the chamber (FIG. 14 ) or which indicates an optical activity of the analyte ( 13 ) indicates. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims, wherein the heating device is implemented as a grid heater. A portable diagnostic system that includes an analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) according to one of the preceding claims and as a power supply having a battery. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) for analyzing an analyte ( 13 ) comprising: a polymerase chain reaction chamber ( 14 ) for processing the analyte ( 13 ); and a grid heater ( 12 . 12 ' . 12''a . 12''b . 12''c . 12''d . 12''e . 12''f ), which is placed in one of the chambers ( 14 ) layer ( 12l ) is integrated and has a two-dimensional geometry, wherein the grid heater is configured according to a two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) having a shape to deliver heat to the chamber, wherein a lateral shape or lateral size of the chamber ( 14 ) to the shape of the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ), the chamber ( 14 ) Is 10% to 30% larger than the two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) such that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid in the chamber at the edge region. An analysis system ( 10 . 10 ' . 10 '' . 10 ''' . 10 '''' . 10 ''''' ) for analyzing an analyte ( 13 ), comprising: a rolling circle amplification chamber ( 14 ) for processing the analyte ( 13 ); and a heating device ( 12 . 12 ' . 12''a . 12''b . 12''c . 12''d . 12''e . 12''f ), which is placed in one of the chambers ( 14 ) layer ( 12l ) is integrated and has a two-dimensional geometry, wherein the heating device is configured according to a two-dimensional temperature distribution characteristic ( 12TC . 12TC ' ) having a shape to deliver heat to the chamber, wherein a lateral shape or lateral size of the chamber ( 14 ) is selected such that a temperature gradient from the center to the edge region of the chamber causes circulation of a fluid in the chamber at the edge region.

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