Method and system for determining biological properties of samples.

[0001] the invention relates to a method for detecting characteristics of samples by means of a micro-panels-reader, wherein said samples are arranged in a matrix array in blisters a microtiter plate, as well as a system for carrying out the method. A preferred field of application is the determination of biological properties of samples.

[0002] In of modern biology sample-multiplex -Method diverse be applied, in which the samples to be tested in a matrix arrangement (wells) in blisters are arranged either sequentially or a microtiter plate ( [...] plate) and there with different, mostly non-contact optical methods of analysis are being analysed. The nature of the analysed samples in this case is extremely varied and can range from homogeneous solutions up to immobilized cells. Particularly the latter, but also other special sample systems ("assays"), need for an optimal reaction or for optimum growth a set temperature as accurately as possible. Modern microplates-readers heating functions therefore have, through which the samples in the microtiter plate can be brought to a defined temperature and kept at this temperature.

[0003] For a conventional systems this temperature control is achieved by means of heating coils with electric current flows, the generate line losses by the ohmic resistance of the heating element due to a temperature radiation. Heat generated by natural air flow by means of these electrical resistance heating Transferring The into the interior of a heating chamber and therefore points on the sample via located there. Partially is used for homogenizing the heat distribution a fan, of the heated air distributed uniformly in the heating chamber. The amount of current, the flows through the heater resistor is regulated by an electric thermal sensor, matches a set desired temperature with the current the readjusts, by means of a thermocouple in accordance with the detected actual temperature and current. In this way, in ideal living cells 37 °C be investigated for example. Furthermore, can be carried out at different temperatures and CONSIDERED-hybridizations can kinetics studies be controlled. Also some require the work at elevated temperature for optimum performance [...].

[0004] The US multi-detection system[...] with heating arrangements for controlling the temperature of samples for 2012/0 300 194 A1 shows universal.

[0005] A task of the invention is to, provide a method and a system for detecting characteristics of samples, the create improved with respect to conventional methods and systems using a tempering of samples investigation possibilities. In particular, the method and the system for determining biological properties of the samples are to be suitable.

[0006] This problem is solved by a method with the features of claim 1, a system with the features of claim 13 and the use of microwave radiation are recited in the dependent claims. Favorable refinements according to claim 26. The texts of all the claims is made by reference to the contents of the description.

[0007] For system is used for heating of samples microwave radiation, before heating, simultaneously with the heating and/or after the heating by means of at least one optical methodology used are to be investigated by measuring the radiation emitted from said sample. Preferably, the samples (some or all) are exclusively by radiation of microwave radiation heated or heated.

[0008] A for carrying out the method has a heating device for heating of samples adequate system, the are located in the blisters a [...]. The heating device comprises at least one microwave source for producing microwave radiation, which for heating on samples can be irradiated. The system needs no further heating devices in addition to the means for generating the microwave radiation, in particular no radiant heating means and no via convection or conduction of heat operating heating device.

[0009] The use of microwave radiation for heating of samples for determining biological and/or other properties of the samples used for this purpose offers specific advantages with regard to previous method and systems. The advantages are, inter alia, in the type of heat generation and the type of heat transfer justified. Since for conventional method (n) only indirectly via the heat the sample reaches the ambient air, the heating is carried out as a rule, relatively slowly (n) of the sample. Even the surroundings of the sample is heated. Conversely, a later cooling can take time to room temperature very long, since a large volume and large area interfaces with a correspondingly high heat capacity (e.g. the walls) by heat radiation, convection and/or by active air transport must be cooled. Due to the slow fading times Warming up and the setting of a precise temperature and/or a temperature control can be difficult.

[0010] In of the claimed invention is used for controlling the temperature of samples and/or microwave radiation is for sample heating. This the efficiency can be increased the sample heating up. Ideally, practically exclusively to that of the sample can be heated, while the surrounding area can be substantially kept at ambient temperature. This can also be as described and it is achieved thereby direct heating that energy directly, does not come into contact with and substantially exclusively in the sample is deposited.

[0011] can further be achieved by use of suitable microwave radiation that the biological and/or chemical integrity of the samples is to be feared is not influenced, in particular because no resonant excitation of individual molecular bonds.

[0012] Since e.g. biological samples usually in low viscosity liquids with permanent dipole moment (in particular in water, aqueous solutions, possibly also in ethanol) are being analysed, it is convenient, to use a radiation for heating, the substantially exclusively interacts with the liquid, which then in turn acts as a direct transfer agent.

[0013] Since in this type of the power transmission are very low, on the one hand, the routes to be bridged, as the heat only needs to be transported within the sample volume, and at the same time the more efficient than in gases impact transmission in condensed phase essential takes place, you get a high degree of efficiency (for example water) to the sample for the energy transfer from the solvent.

[0014] the term "microwave radiation" denotes in this notification 1 GHz to 300 GHz electromagnetic radiation from the frequency range of about, what wavelengths corresponds to 300 mm to 1 mm approximately. In the context of the claimed invention are used preferably 23-cm-band wavelengths of so-called, 2320 and 2450 MHz i.e. wavelengths between, for heating of samples to be tested. 2.45 GHz microwave radiation can be used at a frequency of about in particular. For these frequencies the depth of penetration is in aqueous or alcoholic media remaining sufficient for a largely uniform heating a sample in the typical format wherein [...]. 900 MHz microwave radiation with frequencies up to approximately Optionally can also be used.

[0015] It may be sufficient, and to use for heating a single microwave radiation source to use. In some process variants is provided that in a heating phase and at least one of them at least intermittently microwave radiation a first microwave source is irradiated simultaneously separate second microwave source. This better management is the spatial field distribution of the microwave radiation in the region of the samples possible. If necessary can be achieved, if any that in time average substantially the same amount of energy is deposited in several or all samples, so that the respectively affected samples are tempered under similar conditions. A selective heating up of individual samples or of sub-sets of the samples is also possible. Two separately controllable microwave sources can also be used alternatively to each other or with a time offset.

[0016] On a few embodiments of the system, the heating device a first microwave transmitter antenna and a microwave transmitter antenna on spatially separated second thereof. As a result, among other things, better management of spatial field distribution and thus an adjustment to the distribution of samples in an [...] possible.

[0017] It is possible, does not change substantially during a heating-up phase that the spatial field distribution. For this purpose the heating device may contain a microwave source or more microwave sources, the shall cooperate with fixed preset control parameters.

[0018] It is also possible that the first and second microwave source are controlled and/or amplitude modulated phase modulated coordinated with one another such that by the first and second microwave source generates a time-dependent varying field distribution of microwave radiation is. In a given time window may, for example, individual samples or sub-groups of samples are heated up spatially links, while other samples not simultaneously be heated. (N) by microwave radiation spatial position of the detected sample can The by means of a time-dependent control are varied. In this way the flexibility can be increased in the predetermination of temperature profiles.

[0019] This variable control of the heating effect can be made to a system that the heating device, and at least one of which has a first microwave source separate second microwave source. A may be configured associated microwave control means that the first and second microwave source can be controlled and/or phase modulated amplitude modulated tunable to one another in such a way that through the first and second microwave source can be produced a time-dependent varying field distribution of microwave radiation.

[0020] In a class of systems the heating device has a substantially sealed heating chamber with a metallic shield on to microwave radiation. The heating chamber can be dimensioned such that one or more can be accommodated within the heating chamber for heating [...]. By the metallic shield can be achieved that the microwave radiation used for heating of samples within the area of the shield and it will not be outwardly penetrates. This lying outside the heating chamber can be protected against microwave radiation components of the system.

[0021] In some variants the heating chamber is a separate chamber from the metering chamber. In these cases, a transfer device can be provided between the heating chamber and the metering chamber [...] for transferring. The heating and the measurement are spatially and temporally separated in these cases. The heating chamber can be provided within a separate heating unit, the as Accessory for microplates-reader can be combined together with this to a system. It is also possible, a heating chamber and a separate metering chamber house in a common housing thereof. A solution offers, inter alia, the possibility of having separate chambers retrofit possibility.

[0022] Under other methods is the measuring chamber as a substantially sealed heating chamber with a metallic shield adapted to microwave radiation. This a combined measurement can-and heating chamber be created. In these cases must not be moved between heating and measuring a [...]. The heating can be connected upstream of the actual measurement, so that the heating is completed, before the measurement begins. It is also possible, to perform at least a portion of the heating or to perform a measurement during a heating-up phase during a measurement.

[0023] Many variants the microwave radiation is so generated that the samples (one or more) are located in the region of the far field of the associated microwave production. However, this is not compulsory. It is also a heating near field range possible in the.

[0024] In some process variants is generated for heating of samples a non-propagating microwave evanescence field, having a spatial extent has allowed, the selective heating up of individual samples. In this can be achieved by an adequate system that the heating device has a with a microwave source coupled near-field contains the waveguide, having a radiant outlet opening, having an effective diameter of less than half the wavelength of the microwave radiation has. For example in the range of about 0.5 mm to about can The diameter 6 mm lie. Geometry is so selected that the microwave radiation can not propagate into the environment from the near-field contains the waveguide, but in the near-field contains the waveguide is reflected back. Only a radiant outlet opening and falls in the region of the evanescent proportion may decouple exponentially with increasing distance from the radiant outlet opening in its field strength from. This the spatial resolution is acted upon with the thus determined exclusively by the size of the radiant outlet opening and microwave radiation area is no longer a diffraction-limited manner. The evanescent microwave field can be used for the local heating a sample.

[0025] For sample temperatures it is as accurate as possible a setting of appropriate, to measure the temperature of samples. Although a contacting measurement is possible by means of sensors, are in contact with the samples and/or the can [...], preferred process variants are sample temperature levels by non-invasive sensing of the temperature of samples for the determination of characterized. Through a contactless measurement of the temperature by the temperature measurement can be avoided even in the case of small sample volumes impairments of the sample. A determining the temperature of individual samples or of sub-sets of samples is possible.

[0026] Preferably, sample temperature levels for controlling the temperature the determined used. This the microwave radiation can be controlled as a function of the values derived therefrom sample temperature levels or.

[0027] On the use of microwave radiation as an energy source for locally sample heating up the heating may take place either of samples. The temperature can, for example, by means of an infrared camera or an infrared diode or a contactlessly engaged infrared diode arraysample temperature levels be regulated on the basis of the measured and optionally. In another wavelength range (infrared) operates was based Since this temperature measurement than the for heat suggestion used microwave radiation, and the temperature measurement is not mutually affect the two types of radiation can take place continuously or intermittently if necessary also during the action upon the sample with microwave radiation. A particularly accurate temperature control which is favoured In this way.

[0028] As already it is mentioned embodiments, where the metering chamber as a substantially sealed heating chamber with a metallic shield is designed for microwave radiation, so that the heating can be effected within the metering chamber. In this case separate measures can be provided, to ensure they cannot be damaged affected that against microwave radiation sensitive components of the measuring means for examining the samples not in their function and/or by the microwave radiation.

[0029] Some measuring devices include, for example, a polychromatic light source, from the spectral proportions of light of the light source for the transmission of an optical suggestion path as excitation light is passed in a measuring position, in which a sample is arranged or can be arranged. emission path is emitted through the sample for the transmission of an optical Furthermore [...] a detector provided. With this type of devices may, for example, fluorescence properties of samples are determined. Some such measuring devices have at least one optical path ( emission pathsuggestion path and/or), by a tight opening in the metallic shield the heatingchamber-led to microwave radiation. Through suitable dimensioning and/or shielding of the opening can be ensured that no microwave radiation from the heating chamber, in the region of the opening, can penetrate to the outside. On a few variants of the optical path leads through a light guide, by a tight opening in the metallic shield the heating chamber is guided to microwave radiation.

[0030] In some embodiments is provided, micro-chemical processes, e.g. simple reaction or separation steps, in a (microplates-reader) with a microwave source multi-label -apparatus to perform as a central power source. The system can have means suitable for this.

[0031] This microchemical processes are not limited to biological systems, but also reactions may include in particular, in which experimental conditions such as the stoichiometry of the reactants, the temperature, the temperature gradient and/or the reaction time are permuted. One embodiment polymerizing reactions provides for the implementation of non-biological samples from the region of the before, since in these kinds of reactions a relatively small number of reactants is used and the specific properties of the polymeric end product be achieved by the variation of the experimental parameters listed. A microwave heating suitable microplates-Reader having in particular, for carrying out these reactions, because by having a plurality of samples can be examined micro disk format parallel, and the combination provides distinctive advantages microwave heating the described above. with the. The possibility of direct persecution of the reaction or the review of the given in microplates-readers reaction result is, since polymer reactions often under varying optical material properties such as the refractive index, the absorption or expire autofluorescence, would invention-measuremicro disk reader integrated optical measuring devices with the detected and can be quantified in a.

[0032] Some process variants are characterized in that the heating of the microwave radiation is used to modify this, in its molecular structure the sample. This is preferably indirectly, by the solvent be heated and by the rise in temperature can modify set processes in motion, the the probe substance its molecular structure. Among other things, it is possible, to use the heating by microwave radiation thereto, heterogeneous sample systems separate into individual components, e.g. by way of directly or indirectly adjacent blisters a [...] between distillation. The via microwave radiation in the sample can be kept low so registered energy that a direct modification of the probe substance by microwave radiation can be avoided.

[0033] such process variants can be made possible by e.g. In the system according to that an attachment via one or more blisters ( sample photographs ) is arranged or condense is such that evaporated liquid on the walls of the attachment and dripping back into the cup or the accommodation of the samples. It is also possible that an attachment via one or more blisters arranged or is ( sample photographs ) is such to condense, so that evaporated liquid is deflected and dripping on the walls of the attachment to be at least partly in a directly or indirectly adjacent accommodation of the samples. The corresponding attachments should not from a (e.g. glass or plastic) are made by microwave radiation warm upable material and may be considered as a component of the system or as an accessory.

[0034] by means of suitably dimensioning the attachments to the relatively small dimensions of blisters [...] be adapted to a can. Should an attachment to a cup which can be placed in its particular an outside diameter slightly smaller than an inner diameter of the cup be inlet portion at its open end, so that the attachment can be inserted in a cup with its inlet portion.

[0035] Additional advantages and aspects of the invention result from the claims and from the following description of preferred embodiments of the invention, are explained below with reference to the Figures the. It show:

Fig. 1 shows schematically components of a system, comprising one of a measuring chamber separate heating chamber;

Fig. 2 shows schematically components of a system, comprising a combined measuring and heating chamber;

Fig. 3 shows schematically components of a system, which comprises a measuring device has, the light guides for guiding emission light from the excitation light in a measuring and heating chamber and of measuring and heating chamber;

Fig. 4 shows schematically an embodiment of a near-field hollow wave-guide intended for the local heating of individual samples;

Fig. 5 shows schematically a system for the local heating of individual samples with a near-field contains the waveguide;

Fig. 6 shows another embodiment of a system with near-field contains the waveguide;

Fig. 7 shows another embodiment of a system with near-field contains the waveguide;

Fig. 8 shows in 8A to 8C different variants of a microchemical process, the individual samples in a heating chamber using microwave radiation for selectively heating is performed; and

Fig. 9 shows in 9A to 9C different variants of another microchemical process, the individual samples in a heating chamber using microwave radiation for selectively heating conducted

leads is.

[0036] are schematically some components of a first embodiment of a system 1 Fig. In 100 for determining biological or other properties of samples shown.

[0037] The samples to be tested are in a rectangular matrix arrangement (wells) in blisters 112 110 arranged sequentially or should there by means of a a rectangular [...] or more different non-contacting and optical methods of analysis be investigated. The commercially available microtiter plate can e.g. of a plastic material, such as polyvinyl chloride or polystyrene, or are made of glass and a number of identical sized blisters in straight rows and columns on points, such as between 6 and 1536 blisters, 12*8 = 96 blisters in the example.

[0038] 100, inter alia, for performing process of fluorescence spectroscopy is the system set up. Besides fluorescence measurements may also other was based measuring methods, for example, luminescence measurements or absorption measurements, are carried out. The are not shown for reasons of clarity, components used for this purpose.

[0039] The system 100 has a substantially block-shaped, which is sealable lighttightly on 120 metering chamber is dimensioned so that at least one such can be received within the metering chamber [...] that in each case one of the samples contained in the blisters can be brought into a measuring position by means of a positioning system. The Positioning system has made a horizontally movable cross-table with integrated lifting device in two dimensions, so that the samples can be positioned in three dimensions.

[0040] The metering chamber are associated with measuring devices, with the aid of samples in at least one operating mode of the system from the samples can be investigated by measurement of emitted radiation.

[0041] Outside the metering chamber is a primary light source in the form of a xenon flash lamp with short electrode spacing 130 disposed. There can act is about a continuous wave lamp also depending on the application. The light source has a broad emission spectrum in the visible spectral range.

[0042] with a planar deflecting mirror 132 133 leads suggestion path An optical from the light source up to a measuring position 134,135 into which a sample to be measured (Dashed in a measuring position represented) by positioning the [...] can be positioned for measurement. The sample is located in a recess (well) of the or a cup [...]. The suggestion path 132 are not depicted in more detail, optical elements of the optical spectral proportions of light of the primary light source than serve the transmission of excitation light in the measuring position. The excitation light is irradiated from above into a cup or in the sample perpendicular.

[0043] a candidate substance in aqueous solution 135 is located in the sample. The candidate substance can be encouraged by means of the excitation light, to emit fluorescent light, which is displaced with respect to the excitation light at lower energies or larger wavelengths. The extent of spectral redshift is specifically for the probe substance.

[0044] (the emitted radiation) comes with a emission light via a The 137 136 to a detector 140 emission path -equipped optical parabolic mirror, as a function of the incident light produces electrical signals, are fed to the an undepicted evaluation unit, to evaluate the emission light for characterizing the sample spectrally. In discussed example comprises a photomultiplier the detector.

[0045] also can be carried out with the system 100 absorption measurements. This the light can be guided through the sample via the optical suggestion path 132, the is located in a microtiter plate with transparent bottom. The penetrating light by passing through the sample to a not-shown light detector shall take, typically a silicon photodiode having wide spectral range.

[0046] The system 100 is also suitable for the measurement of luminescence emission, the chemically, biologically or biochemically is generated. Not shown are injected via injectors initiator substances which can To this end, the light emission from initiating in a sample. The light emitted via the parabolic mirror is guided via the optical detector 140 137 136 for emission path.

[0047] To the system 190 100 belongs a heating device, with which the samples can be heated by means of microwave radiation [...] held in a. To of the heating device 150 190 120 separate heating chamber from the measuring chamber a belongs, the addition to the metering chamber, and at a distance therefrom is arranged at the same height within the housing 105 of the system. A chamber wall encloses a substantially cuboid interior 152 155 of the heating chamber is dimensioned so the fits that at least one completely [...]. On the inside of the chamber wall is a metallic shield 154 mounted so that the inner space is sealed with respect to the delivery of microwave radiation 152 from the environment. Protecting 154 can have joined metal sheets without a gap for example, to the inner sides of the chamber walls are fixed.

[0048] a first microwave transmitter antenna 152 is located in the interior 162-1, 160-1 is connected to a first microwave source 152 into the interior space and capable of radiating microwave radiation from this generated. A optional second microwave transmitter antenna is spatially separate from the first microwave transmitter antenna 162-2 162-1 is excited, for example, arranged on the opposite side and into the interior space 160-2 for applying microwave irradiation via a second microwave source. The 160-1 microwave sources, are 160-170 connected to a control device 2 that is amplitude modulated is configured in such a manner that the first and second microwave source can be controlled independently of one another and/or phase modulated.

[0049] In the microwave sources it is for semiconductor-based microwave sources in the discussed example, 2.45 GHz and the microwave radiation with a frequency of from about 50W can produce a power rating in the range of about. Alternatively, useful power and frequency would be appropriate for example, take a magnetron.

[0050] To 190 180 belongs to the non-contact temperature measurement of the heating device a temperature sensing device by detecting and evaluating infrared radiation from samples. This sample temperature levels can be determined, on the basis of the heating device can be controlled, so as to achieve a temperature control. In the discussed example 182 comprises an infrared diode array 180 temperature sensing device The, which [...] outside the shield mounted above the receptacle for the is connected to the control device and 170. In the shield are small holes introduced in the region of the infrared diode array, through which the infrared radiation can pass, while the microwave radiation with a longer wavelength is blocked.

[0051] For the transfer between the heating chamber 150 and the metering chamber 120 is a microtiter plate 175 provided a transfer unit, the 170 of the system can be controlled via the [...]. The transfer unit has a on guide rails horizontally movable panel receptacle 176 on, each of which can receive a single microtiter plate. The panel receptacle has a vertically displaceable frame 177 on, [...] with two opposing edges may rest on which the. The [...] are transferred to other components can thereby, e.g. by settling on a pedestal in the heating chamber. The 175 also has a fine positioning of the received transfer unit drives for on microtiter plate, so that a is present disk manipulator, both the transfer and the fine positioning can accept. The transfer unit can thus form a common unit with the cross-table Sample setting.

[0052] Samples shall be heated exclusively by means of microwave radiation. Due to the dipole moment of the water molecules in the lower GHz-region causes a irradiation with electromagnetic radiation (corresponding to wavelengths in the range of a few centimetres), so-called microwave radiation, an action of force on the water molecule, so that a torque acts on the water molecule and forces it to a rotational movement. Friction-with adjacent water molecules heat is produced, this consideration important However, in that the employed water moleculerotation resonant frequency of the radiating frequency must not correspond to a.

[0053] help of the arrangement can be ensured that the heating or the incubation of samples in an independent, metallic-conductive enclosed space takes place (interior 152), and thus represents a cavity for the microwave excitation corresponds to the one closed on all sides contains the waveguide. A [...] with the samples to be tested by means of the movable panel receptacle 176 is retracted into this cavity and can be disposed in a heating position there. The transfer unit can then leave the heating chamber. The interior 152 can then also be sealed by means of a shielded Drawer radiation outwardly. A maximum of 1 mm wide and at least 1 cm are of the drawer Am circumference long column, are impermeable for the inside excited microwave radiation. The sample drawer can be realized, for example, by a rear bezel to said column. With such an arrangement it is achieved prevail, 152 reproducible and controllable electromagnetic conditions in the interior space and that the spread of microwave radiation by the microwaves-transmit antennas to the samples depends not further during the development of the device, since, for example by electrically conductive components are excluded field bucklings possible. The heating device can be designed as an independent assembly 190, in addition to a metering chamber can be integrated in suitable dimensioned devices.

[0054] A homogeneous field distribution within the cavity can be achieved at least in the time average, by said two discrete, 162-1.162-2 microwave transmit antennas in the interior placed shall be controlled in amplitude and phase that if possible no radiant hot-spots arise, or these in time over the microtiter plate to move and thus ensure a homogeneous energy input (in time average).

[0055] After the transfer of the microtiter plate with the heated samples in the metering chamber can be carried out in a manner known per se there radiation measurements, e.g. fluorescence measurements.

[0056] In Fig. are some components of a second embodiment of a system 200 schematically 2 biological properties of samples shown for determining. Through a system similar to as in the first embodiment can also be carried out on samples luminescence measurements or absorption measurements and fluorescence measurements, are housed in a cup 210 [...], [...] of the first embodiment may be identical or similar to said.

[0057] A feature of the system 200 is, at the same time as the heating chamber 220 290 that the lighttightly is designed a heating device which is sealable metering chamber, so that they can serve as a combined measuring and heating chamber, the allows, by means of microwave radiation to heat the samples located in the metering chamber or heat. This heating can. Temperature adjustment and measurement are carried out simultaneously if necessary.

[0058] The substantially cuboidal Measuring and heating chamber as the heating chamber can be dimensioned similar to or equal to 120 of the first embodiment. To encloses can serve to achieve is provided that the measuring chamber also as a substantially tight heating chamber to microwave radiation, a metallic shield only partly represented, the the interior 252 of the measuring chamber substantially completely in such a way in the closed metering chamber that microwave radiation from the interior of the measuring chamber or not. Heating chamber, can penetrate to the outside. To build the metallic shield can be mounted on the interior walls of the metering chamber metal sheets, are joined together at the joint locations without a gap or with very narrow gaps.

[0059] At least one part of the metallic shield is formed by perforated plates 255-1.255-2, the means of a regular pattern of holes is have, their diameter in the range of 1 mm to 30 mm, in particular in the range from 2 mm to 20 mm, and thus is so small that microwave radiation cannot penetrate through these perforated plates. The holes, however, makes it, passing through the metallic shield optical paths of measuring devices.

[0060] The measuring equipment, the 220 are associated with the metering chamber, a primary light source arranged outside the metering chamber comprise 230, the outputs have a continuous emission spectrum in the visible spectral range. A suggestion path 232 leads through an aperture (hole) in the lateral optical 255-1 board via a perforated plate in the interior of the metering chamber arranged flat deflecting mirror from the light source 230 to 234 to said measurement position, in which a sample to be measured can be positioned. The (the emission light) radiation emitted from the sample excited in such a way with a parabolic mirror passes over a detector disposed outside the metering chamber 240 236 to a optical emission path equipped, the comprises a photomultiplier. The emission path leads through holes or. Openings in the perforated plate 255-2 is mounted on the chamber inner wall above the measuring position soft.

[0061] Those components of the heater 290, which are provided for the irradiation of the microwave radiation to the samples, can be identical or similar corresponding components of the first embodiment, to the description of the first embodiment is cited why in so far. Here there are two independently controllable microwave transmitter antennas 262-1 also, 262-2, each of which are arranged within the metallic shield, urn can be controlled to generate microwave radiation directed to the samples, and by way of a control device with respect to amplitude and phase so that in the region of the samples the desired field distribution for heating the samples.

[0062] also correspond to those of the first embodiment can the components of the contactlessly operating temperature measuring device, which is why its description is cited. In particular is also here an infrared diode array for non-contact measurement of the temperature of the samples above the receiving position for 282 [...] outside the shield (perforated plate 255-2) mounted the. The measurement is performed by passing holes of the perforated sheet metal.

[0063] some components of a third embodiment of a system 3 are schematically In Fig. 300 for determining biological properties of samples shown. Again, the metering chamber is 320 390 designed simultaneously as a heating chamber of a heating device, the allows, by means of microwave radiation to heat the samples located in the metering chamber or anneal. The heating device with at least one microwave transmitter antenna can be built up in the interior of the metering chamber 360-1 as the heating means of the preceding embodiments identical or similar.

[0064] differences to the second embodiment remain primarily in the manner, as the optical paths of the metering chamber are designed associated measuring device. The suggestion path 332,334 330 for measuring position from the light source disposed outside the metering chamber leads, leads 333 by a first light guide, the leads through a hole in the perforated plate arranged above the measuring position 355-2 of said metallic shield. The emission path 336, in which radiation emitted from the measurement position detector disposed outside of the measuring chamber is guided to the 340, runs 337 by a second light guide, which can also be passed through a hole in the perforated plate arranged above the measuring position is guided 355-2.

[0065] In this embodiment is therefore a common chamber for incubation and measurement combined with measuring devices, the light guides from the light source to the sample posture shifting method for detector have sample position and of. figurativy is represented a variant, in which the light guides are designed as y-Split, so that side by side and are guided suggestion pathemission path in sections. It is also possible, in such a way on the optical fibers for the execute suggestion pathemission path coaxially that both paths can be guided through the same through-passage within the metallic shield. The 3 shows a cross-section through a detail in Fig. coaxial fiber bundle 350, in which a central optical fiber and the about the central optical fiber leads emission path the light from the exterior are provided around optical fibers for guiding of the excitation light. It is also possible, in the reverse arrangement to use the coaxial fiber bundle, so that a central optical fiber leads the excitation light, while the emission light is guided by the outer optical fibers arranged about the central optical fiber. The allocation of optical fibres, the excitation or emission light are acted upon with, can also be arbitrary, so that a homogeneous mixing of excitation and emission optical fibers it is achieved.

[0066] If terminates in the interior of the heating chamber a light guide, the ferrule should ideally consist of plastic or another electrically non-conductive material of the light guide, so as not to disrupt the microwave field distribution. Metallic ferrules are also conceivable, but in the interpretation of the heating chamber and then should the calculations to optimize the field distribution of the microwave radiation are involved.

[0067] With graphically displayed must be introduced in each case the heating device has a metallic shield with embodiments equipped heating chamber, with samples in the at least one [...], to heat the samples by means of microwave radiation. Samples far field range the microwave transmit antennas are located in the.

[0068] It chamber-free systems are also possible that is, systems without separate heating chamber for containing a [...]. The described below the microwave radiation to a non-propagating evanescence field systems exploit sample heating. 4 shows schematically for purposes of illustration, an embodiment of a near-field waveguide Fig. and 5 schematically a system 400 usable therefor Fig. 500, which is equipped with such a near-field contains the waveguide for sample heating.

[0069] When [...] -microwave excitation is by means of a microwave source in a manner similar to other embodiments the microwave radiation is first generated and via an antenna 460 410 400 radiated into the interior of the metallic near-field waveguide. In the exemplary embodiment, the near-field contains the waveguide is tapered at its end opposite to the antenna impedance matching and terminates in a round or angular exponentially for a continuous radiant outlet opening 420, whose effective diameter is significantly smaller than the wavelength of the microwave radiation D. For example in the range of 0.5 to 3 cm can The diameter lie. The microwaves can propagate into the far field not here, but it arises immediately before the radiant outlet opening a in its field strength exponentially from fading away evanescence field, the can be used for the local heating a sample.

[0070] In a not graphically displayed embodiment, the near-field contains the waveguide on a geometry, through which the reflected microwave radiation is not reflected back to the source, but passes to it. The above-mentioned process can either by a suitable deflection of the microwave radiation are repeated several times, such that a plurality of samples can be simultaneously heated in said cup a [...]. By lateral Method [...] relative to the radiant outlet openings, it is thus possible to all the blisters of the successively with [...] act on microwave radiation.

[0071] In Fig. 5 is a schematic of a chamber-free system 500 with microwave [...] and beam guide Sample Application via light guides of the measurement system represented. The [...] 510 is located within a not-shown metering chamber of the system in this case, the no internal metallic shield against the escape of microwave radiation or needed. There can constitute the measuring chamber of a conventional system act. 530 532 for measuring position coming from the light source over the suggestion path The excitation light is irradiated from above into the sample, the emitted radiation emits upwardly, 540 536 to the detector by the passes emission path.

[0072] The heater 400 500 590 of the system comprises at least one near-field contains the waveguide, the discussed example is arranged below the holding plane of the so [...] is located in that the upwardly directed outlet opening 420 in a plane immediately below that level, in which the trays are arranged [...] 512 of the blisters. The distance between and bottom of a cup is sized radiant outlet opening that from the radiant outlet opening exiting from below through the material of the microwave evanescence field into the aqueous sample [...] penetrate and can heat this.

[0073] as small in diameter is advantageously the radiant outlet opening that in each case only a single sample by means of microwave radiation [...] thus typical can be heated, while the other samples practically of microwave radiation to ambient temperature (or any other preset temperature) remain unaffected. A selective heating of individual samples is thus selectively possible. The heating can be monitored and regulated by the contactlessly operating temperature sensing device under observation. This has, in a manner similar to other embodiments, an infrared sensor, disposed above the receiving position for the aid of a three-dimensionally displaceable [...]sample manipulation mechanism 582 510 575 for moving the. relative to the radiant outlet opening 420 can individual samples microtiter plate before the outlet opening be positioned successively..

[0074] 6 and 7 700 600 in Fig. in Fig. systems In the are the means for positioning the [...], for heating of samples using near-field contains the waveguide 4 and 5 and for temperature control identical to the embodiment of the. differences here only with respect to the body Fig. of the measuring means.

[0075] When system 630 600 of the light without the use of the light source 6 is irradiated Fig. light guides via an inclined deflecting mirror from above onto the sample, while the emission light passes to the detector via a parabolic mirror (see item for example first embodiment).

[0076] When variant of 732 736 emission pathsuggestion path are separated and the 7 Fig. via a geometric the 770 in the form of a beam splitter hole mirror, and a central opening is disposed above the measuring position of the has, through which the excitation light directly from the light source disposed above the measuring position in a sample can be irradiated 730. Reflective surface of the geometric beam splitter to the detector 740 passes viaa emission light The.

[0077] Fig. 8A shows an embodiment for a configuration for heating a sample in a boiling point of the solvent used for a longer period to a maximum of the [...]. 810 812 a [...] is substantially gas-tight on a cup 820 brought an attachment, the essentially consists of an upwardly open pipe of a non-metallic material, e.g. carbon fiber-interspersed plastic or glass.

[0078] The external diameter of the attachment is on a cup which can be placed in its slightly smaller the internal diameter of said cup 812 at its upper open ended inlet portion, so that the attachment can be inserted with its inlet portion a small piece to said cup. Patterns are projected at the inlet portion is outwardly projecting shoulder formed a, whose outer diameter is a little larger than the diameter of the cell opening, so that said attachment said cup seals substantially gas-tight.

[0079] By heating of the specimen 835 by means of microwave radiation from a lower temperature to a higher temperature T Ti₂>T-i according to one of the above-described method is increasing as the steam begins to boil the solvent upwardly towards the inside of the attachment and. Since by the use of microwave radiation as an energy source is substantially not heated the upset tubing, cools the steam to the tube walls and is condensed. The liquid condensate can drip back into the receptacle is 812.

[0080] 8B Fig. In another embodiment, the tube is such accordance that it tapers upwards. Through this arrangement is achieved a more efficient [...] of the solvent vapor.

[0081] In another embodiment, the wall thickness increasing upwards in Fig. 8C [...] has a shown. Due to the high heat capacity of the tube reaches a larger thermal gradient is This of T₂according to a more efficient and thus at a given length of tubing T-i gas condensation.

[0082] Fig. [...] 9A shows an embodiment for the separation of liquid substance mixtures in. Here 812-1 two adjacent blisters are, 812-2 830 connected to each other via a bridge to the second cup sloping down towards, such that the bridge on the first cup 812-1, in which the mixture of substances is located and can be described as distilling cell, sits substantially gas-tight, while supported on the second cup 812-2, can be described as the can rests collecting main cell takes place, so that a gas exchange with the environment. By heating of the sample with microwave radiation and begins to boil the solvent according to one of the above-described method as steam rises upwardly. Since by the use of microwave radiation as an energy source is substantially not heated the fitted bridge, cools and condenses the steam to the is bridge walls ab. The condensate flows in order to arrive at a trailing side of the bridge from the collecting main cell. Taking advantage of different boiling points can be separated from one another in this way two fractions substance mixture a. The separation efficiency can be enhanced, by inserting into the bridge has webs not shown, the project into the gas space above the distilling cell. At these webs of the steam can [...], drop and re-enter the gas phase.

[0083] In this way increases the number of effective distillation steps and the separation efficiency is increased. In of classical preparative chemistry this method is referred to as the increase in the number of theoretical plates ( column distillation ) by introducing a distillation column.

[0084] in the embodiment shown, the bridge has a 9B Fig. An article in collecting main cell rising wall thickness to direction. Due to the high heat capacity of the bridge is achieved a larger thermal gradient This of T₂according to a more efficient and thus at a given T-i bridge lengthgas condensation.

[0085] Another 9C illustrated embodiment used as a bridge in a curved tube Fig., comprising a substantially horizontal opening distilling cell has on the side of the back and on the side of the to the gas volume collecting main cell a substantially vertical opening, so that dripping collecting main cell into the condensed gas in the tube.

[0086] Both [...] with microwave radiation for heating samples in procedures set out above, in the modification of the molecular structure or for the separation of a homogeneous substance mixture is not so the microwave radiation used that immediately by the application of the microwave radiation is influenced the molecular integrity of the sample. Rather, serves only the microwave radiation, to heat the solvent by orientation polarisation of the dipoles contained therein. This [...] is that the obtained findings also for larger Resize substance quantities.

[0087] Current Research investigate, intensive microwave radiation can directly influence the extent to which the chemical properties of substances. This influence is not provided in the described examples, can be used but, if necessary, in addition, when high-power microwave sources are used.

[0088] embodiments of the claimed inventions may offer one or more of the following advantages.

[0089] A exclusively local heating of the sample: Thereby a shorter heating times be achieved in the range a few seconds. equipment interior is not heated and must therefore not cool the At the same time slowly. Therefore measurements can be carried out at different temperatures, in a direct sequence.

[0090] A exact temperature control: Will prosecuted via an IR diode or another IR sensor the temperature of the sample in real time, by adjusting the microwave power this may (in the cw-or in pulse mode) via a feedback (feedback control) to be regulated very precisely. Therefore you would achieve a high precision [...], the not be applied in a similar way to conventional methods is possible.

[0091] A generation of temperature profiles: A active temperature-control loop in conjunction with the short reaction times therefor can ensure that user specific temperature profiles can be driven. The kinetics studies of advantage, on the one hand, e.g. for two-dimensional and is opened on the other hand the field of the DNA hybridization: cooling profilehybridizing error by a clear definition of the heating and can almost be avoided.

[0092] Enlarging the application scope of the devices. It is used to call the inventor possible, within the framework in a simple reaction and separation steps u-chemical processes multi-label -device with a microwave source to perform as a central power source. In combination with injectors or with corresponding attachments can also here dispersion units and stirred and heated under reflux in a distilled neighbour-wave be from a well. It will be essential that a large temperature gradient is, the is reached, by adding a (e.g as a simple bent tube carried out) is heating the distillation bridge prevented. The local heating by means of microwave radiation satisfies this requirement.

[0093] In this way can a multi-label reader or itself. Microplates-reader from the pure Gauge for integrated develop synthesis robot with analysis unit. Such an approach is extremely helpful for systemic approaches polymer research e.g. in the, since here with a limited number of starting materials, reaction conditions and concentrations through-permutated be, so as to achieve desired product properties.