Title of Invention

METHOD AND DEVICE FOR RECORDING PROCESS PARAMETERS OF REACTION FLUIDS IN SEVERAL AGITATED MICROREACTORS

Abstract

The invention relates to a method for recording process parameters of reaction liquids in a plurality of microreactors (10, 19) of a microtitre plate, which are continuously shaken at least until the end of the reaction in all of the microreactors by means of a rotation shaker, wherein (a) the process parameters in the microreactors are recorded during the reaction whilst shaking continuously, (b) the process parameters are recorded with the aid of a sensor optics device (17) comprising a sensor (15) and a radiation source (12), (c) the process parameters in the individual microreactors (10, 19) are recorded successively by a sensor optics device (17) disposed on a positioning unit (7), (d) the electromagnetic radiation from the radiation source (12) is successively introduced into the reaction liquid of the microreactors of the microtitre plate and the sensor (15) associated with the radiation source records the electromagnetic radiation emanating from the reaction liquid of the microreactor, (e) after recording the process parameters in one of the microreactors, the sensor optics device (17) is moved by the positioning unit (7) to another microreactor, characterised in (f) that the sensor optics device (17) aligned under one of the microreactors is not moved at least during the recording of the process parameters so that shaken microreactors (10, 19) move relative to the sensor optics device (17), (g) wherein the shaking diameter of the rotation shaker is selected to be smaller than the diameter of the bottom (10) of the microreactors so that during the recording of the process parameters in one of the microreactors the electromagnetic radiation of the sensor optics device (17) is introduced exclusively into this microreactor and the electromagnetic radiation emanating from each microreactor following the introduced electromagnetic radiation is exclusively recorded by the sensor (15) of the sensor optics device (17) associated with the introducing radiation source (12). The invention also relates to an apparatus for carrying out the method as claimed.

Full Text

The invention relates to a method and an apparatus for recording of process parameters of reaction liquids in a plurality of microreactors, which are shaken continuously at least until the end of the reaction in all of the microreactors, with the process parameters being recorded in the microreactors during the reaction with the aid of at least one sensor optics device which introduces electromagnetic radiation from a radiation source into the reaction liquid in a microreactor, and with electromagnetic radiation which originates from the reaction liquid in the microreactor being recorded by a sensor which is associated with the radiation source. The invention is particularly suitable for automated recording of process parameters of microbial, biochemical, enzymatic and chemical reactions in reaction liquids which are shaken without interruption until completion of the reaction in all of the microreactors. By way of example, the biomass, substrate, product and byproduct concentrations, the self-fluorescence of cells, the fluorescence of fluorescent proteins and amino acids, pH, T, pO2 and pCO2 values, the oxygen transfer rate (OTR) and the carbon-dioxide transfer rate (CTR) can be recorded as parameters of the reaction liquids. In particular, the above-mentioned parameters are intended to be determined in microreactors with a size of 10 µl-5ml, in order to reduce the expense for chemical, biochemical, enzymatic and microbial optimization methods, also referred to as screening. Screening is considered, for example, for the core-area selection, media optimization and optimization of process control. The small volumes in the microreactors allow the required high throughputs in many areas of research and development, in particular such as combinational chemistry and molecular biotechnology. So-called microtiterplate readers are known from the prior art, for recording the absorption and fluorescence in microbial reaction liquids. The shaking movement of the microtiterplates must be interrupted for each recording of process parameters during the reaction. The greater the number of process parameters that are intended to be obtained while the reactions are taking place, the more frequently the shaking movement must be interrupted, thus disturbing mixing processes and substance transport processes. This can result in anaerobic conditions in the case of cultures which cause greater or lesser damage to the various microorganisms. An absorption microtiterplate reader such as this for 200-well microtiterplates for monitoring of microbial growth is available, for example, from the Thermo Electron Corporation, Waltham, MA, USA. The light absorption by the cells located in the wells is recorded. For this purpose, electromagnetic radiation from a radiation source is introduced into the reaction liquid in the wells, and the electromagnetic radiation emitted from the reaction liquid in the microreactor is recorded by means of a sensor. The sensor signals depend on the layer thickness passed through and on the cell concentration. Furthermore, US 6,673,532 D2 has already disclosed a microtiterplate reader for recording of the absorption in microbial culture liquids, in which the shaking movement of the microtiterplates need not be interrupted in order to record the absorption during the reaction. By way of example, the known apparatus comprises a microtiterplate with 96 wells, which is held by a microreactor platform. The individual wells have a volume of between 100 ul and 250 ul. At least one sensor optics device is located in a sub- platform arranged under the microreactor platform and has an excitation source, for example a light-emitting diode, as well as a detector, which records the absorption of the electromagnetic radiation from the excitation source in the reaction liquid in the microreactor (wells). The change in the measured absorption indicates a change in the concentration of the analyte in the microreactor. One refinement of the reader provides for the LEDs and the detectors to be moved from one microreactor to another by means of a robot. Another refinement provides for each microreactor to have at least one associated LED and one associated detector within the sub-platform. The sub- platform with the sensor optics device or devices is once again located on a shaking apparatus, which is mounted on a positioning table. The shaking apparatus is a specially manufactured device, in order to allow integration between the positioning table and the sub-platform. The positioning table can be moved in the XY axis direction and is used for the purpose of moving individual microreactors under a dispenser. By way of example, this known apparatus can be used to assess growth conditions for the microorganisms in culture liquids in a valid form, since it avoids the problems which occur as a result of interruption of the shaking movement. However, the apparatus has the disadvantage of its complex design, particularly that of the shaker, which is specifically matched to the apparatus. A further disadvantage is that the sensor optics device or devices are also shaken in the sub-platform. Owing to the high shaking frequencies and the centrifugal forces associated with them it is possible for problems, and thus errors, to occur in the recording of the process parameters which, in some circumstances, make it necessary to repeat the reaction. This can result in undesirable delays, particularly in the case of series of investigations relating to microbial culture liquids, enzymatic and chemical reactions. Furthermore, WO 92/10754 A has already disclosed liquids in microtiterplates being analyzed during the shaking process. This analysis is carried out using video cameras, photodiodes and a scanning device. Finally, US-A 5,595 708 discloses a sensor optics device for series testing of a large number of reactors which are arranged in a plurality of tilting frames. The sensor optics device is arranged on an XY positioning unit, which moves to the individual reactors successively. The signals from the sensor optics device are processed further in a digital evaluation unit. The tilting movement of the reactors is interrupted during the measurements. Against the background of this prior art, the invention is based on the object of specifying a method for recording of process parameters of reaction liquids in a plurality of microreactors which are shaken continuously during the reaction, and which method operates reliably. A further aim is to specify an apparatus for carrying out the method, which can be provided largely by the use of standard components and apparatuses from biotechnology. This object is achieved by means of a method having the features of the independent claims 1, 2, 4 as well as an apparatus having the features of the independent claim 12. The advantages and effects of the various solutions will become evident from the following description. All of the solutions include the features that each sensor optics device is not moved, at least during the recording of the process parameters, so that the shaken microreactors move relative to each sensor optics device, and the electromagnetic radiation which originates from each sensor optics device is introduced, durirg the recording of the process parameters in one of the microreactors, exclusively into this microreactor, and the radiation emerging from the reaction liquid strikes only the sensor of the sensor optics device. These features are based on the discovery that continuous recording of the process parameters is possible ever when each sensor optics device which is arranged in a positioning unit has not been moved during the recording of the values of a process parameter, for example during the recording of an instantaneous value of natural fluorescence during the ongoing reaction. The relative movement which occurs during this process between the shaken microreactors and each sensor optics device is without any problems provided that the electromagnetic radiation of each sensor optics device is introduced, during the recording of the process parameters in one of the microreactors, exclusively into this microreactor, and that the radiation which originates from the reaction liquid strikes only the sensor of the associated sensor optics device. The biomass concentration in one of the microreactors may, for example, be recorded with the aid of scattered light striking the sensor of the sensor optics system, or the radiation which originates from the natural fluorescence of the cells. The substrate concentration, product concentration and byproduct concentration can be trapped with the aid of IR or Raman spectroscopy. Biotechnological substrates such as glucose or glycerin are IR active and Raman active, and form a specific spectrum, which can be detected even in very complex media. Metabolism byproducts such as acetic acid and ethanol likewise have a characteristic spectrum. Organic substrates can be detected in the microreactors by transmission of the spectrum via optical waveguides to an IR spectrometer or Raman spectrometer. More detailed statements can be found in Sivakesava S., Irudayaraj J., Ali D. (2001): Simultaneous determination of multiple components in lactic acid fermentation using FT-MIR, NIR, and FT-Raman spectroscopic techniques, Process Biochemistry 37, 371-378. In one advantageous refinement of the invention, the reaction liquids in the microreactors have at least one chemical sensor material, in particular such as a fluorescent dye which, in particular, can be immobilized on at least one inner wall of the microreactor. Fluorescent dyes react specifically to their environmental conditions. For example, platinum porphyrins or ruthenium complexes react to the presence of oxygen by cancellation of their fluorescence characteristics. Fluorescence indicator solutions likewise have a sensitive reaction to changes in the dissolved CO2 concentration (pCO2) and of the pH value by changing their fluorescence characteristics. If these substances are immobilized in a porous polymer matrix or are dissolved or suspended in the reaction liquid, then they act as optical sensors (also referred to as optodes) for ;pH, T, pO2, pCo2 values (Liebsch (2000): Time Resolved Luminescence Lifetime Imaging with Optical Chemical Sensors, Dissertation at Regensburg University). However, it is also possible to use normally dissolved pH indicators. .Brief description of the accompabymg drawings. The invention will be explained in more detail in the following text with reference to the figures, in which: figures 1A-C show three variants of preferred apparatuses for carrying out the method according to the invention; figures 2A-C show various embodiments of a sensor optics device for the apparatus according to the invention; figures 3A-B show two schematic plan views of a cylindrical microreactor which rotates with a fixed shaking diameter over a light beam of a flashlamp; figure 4 shows an illustration in order to explain a first variant of the method according to the invention; figure 5 shows an illustration in order to explain a second variant of the method according to the invention; figure 6 shows an illustration in order to explain a third variant of the method according to the invention using chemical sensors, and figure 7 shows measurement curves of a culture of Hansenula polymorpha A microtiterplate (1) with an array of microreactors is inserted into a holder in a tray (4) of a rotation shaker (5, 6). The rotation shaker (5, 6) reaches a maximum shaking frequency of, for example 500-2000 urn. The shaking diameter of the rotation shaker is matched to the recording process. Electromagnetic radiation at a wavelength of 200 nm-25 µm is introduced via a sensor optics device (17) through the cutout (11) in the tray (4) into those wells of the microtiterplate (1) which are permeable to the radiation, and the radiation emerging from the wells is recorded by the sensor optics device (17). Figure 1A shows one embodiment with a stationary rotation shaker (5) , in which the subarea of the tray (4) on which the microtiterplate (1) is fitted projects beyond the drive. This ensures free access between the sensor optics device (17), which is arranged on an X/Y positioning unit (7) , and the microtiterplate (1) from its lower face. Figure 1B shows an embodiment with a tray (4) which is driven by a suspended rotation shaker (6) . That part of the tray (4) to which the microtiterplate (1) is fitted need project beyond the rotation shaker (6) in this embodiment only when the wells are also intended to be accessible from above. Figure 1C shows an embodiment in which the microtiterplate (1) is held by a shaking frame (16) with a cutout (11). The shaking frame (16) is separated from the microtiterplate (1) in the vertical direction by the planar tray (4) . Bscause the shaking frame (16) is designed to be open at the side, the sensor optics device (17) can be moved under each well in the microtiterplate by means of the X/Y positioning unit (7) without being impeded from the side. Figures 2A, 2B show apparatuses in which the electromagnetic radiation from the radiation source and the radiation originating from the reaction liquids are passed via optical waveguides (2, 25) . The variant in figure 2A shows the radiation being carried separately via two optical waveguides (2) . The radiation is introduced via one optical waveguide (2) which leads towards the reactor, and the radiation originating from the microreactor is carried via an optical waveguide (2) which leads away from the microreactor. A radiation source that feeds the sensor optics device, and a sensor, are located in a reader (3) . The variant shown in figure 2B differs from the variant shown in figure 2A only in that the two optical waveguides (2) are combined in one Y-optical waveguide (25) . The optical waveguides are composed of individual optical fibers or else of optical fiber bundles. The sensor and the radiation source which feeds the sensor optics device are located in the reader (3). Figure 2C shows a sensor optics device (17) without any optical waveguides. The radiation source (12) and the sensor (15) are located directly in the sensor optics device (17), which may additionally have filters 13, 14 in the beam path. The excitation light from the radiation source (13) is aligned directly with each individual microreactor, while the reaction-dependent radiation which originates from the reaction liquid in the microreactor strikes the sensor (15). The sensor optics device (17) is connected to an electronic circuit (9) via a cable or cables (26) in order to supply the radiation source (12) and in order to transmit the sensor signals. The circuit (9) controls the radiation source (12) and is used for reading the sensor signals. In all variants, the data is acquired and evaluated by means of a data processing unit, for example a computer (8) . In the variants shown in Figures 2A, B, the functionality of the electronic circuit (9) is integrated in the reader (3) . The sensor optics device (17) is attached to the arm of an X-Y positioning unit (7), in all of the variants. The X-Y positioning unit (7) is likewise driven by the computer (8), by means of a control software. Depending on the requirement for the measurement task, it is possible to inject electromagnetic radiation in a closely constrained wavelength range, for example produced via optical filters 13, diffraction gratings, prisms or directly by a radiation source with a defined spectrum, such as a laser or an LED, and to supply the sensor only with specific wavelengths from the emitted light. The emitted light can likewise be filtered by means of optical filters 14, diffraction gratings or prisms for this purpose. If a flashlamp is used to excite the analytes in the reaction liquid of a microreactor, it is advantageous for the shaking rotation speed of the rotation shaker (5, 6) and the pulse repetition frequency of the flashlamp to be matched such that no beat states occur. Beat states occur when the light flashes strike a small number of positions on the bottom of the reactor and the positions move on the bottom of the reactor as a result of lack of synchronization between the shaker frequency and the flash frequency. The microreactor which is illustrated in figure 4 and is bounded by a circular-cylindrical bottom (10) and a cylindrical casing (19) , rotates with a fixed shaking diameter over a fixed-position light beam (18) of the sensor optics device (17). The rotation of the microreactor results in the light beam (18) describing a circular line (11) on the bottom (10) of the microreactor (see figures 3A and 3B). Figure 3A illustrates a state in which only 3 flashes are distributed over the circumference of the circular line (31) , as a result of the choice of the shaking rotation speed (n) and of the flash frequency (fa) of the flashlamp. In this case, fa = 3*n, so that only 3 flashes are initiated per revolution. Beat states occur particularly if the flash frequency (fa) is a natural multiple of the shaking rotation speed (n) . In this case, the light flashes initially start at the same positions (32) on the circular line (31) . Ef the coverage of the circular line (31) with flashes is low ( not synchronized with the liquid movement in the reactor, the positions (32) move in the clockwise sense or counterclockwise sense on the circular line (31) . Because of the angle (22) of the sensor optics device (17) , this leads to different intensities of the electromagnetic radiation emitted from the microreactor, and disadvantageous oscillations of the measurement signal can thus occur. A greater number of light flashes is therefore desirable in order to avoid beat states, with these being distributed uniformly over the circular line 31. This can be achieved by satisfying the condition fa = N*n (where N = natural number) . Figure 3B illustrates one example relating to this. Thirteen light flashes are shown over the circumference of the circular line (31) . The light flashes can be emitted during one revolution, or else during a plurality of revolutions, of the rotation shaker. The relationship n=fa*P/U (where P = number of flashes on the circular line and U = number of revolutions before P is reached) results in the operational conditions being set such that as high a number of flashes (P > 10) as possible are distributed over the circumference of the circular line. This makes it possible to obtain a stable and homogeneous sensor signal in order to record the process parameters in the microreactor. A further possible way to stabilize the sensor signal is to synchronize the flashes from the radiation source with the shaking drive, and thus with the liquid movement in the microreactors. The position of the tray (4) with respect to the sensor optics device (17) can be determined at any time by means of a position sensor (for example a light barrier, an acceleration sensor or a Hall sensor). The light flashes from the flashlamp are triggered taking into account the position information. The flash is preferably triggered when the reaction liquid which is sloshing in the centrifugal acceleration direction is located above the sensor optics device (17). This prevents the light beam (18) from striking zones of the microreactor in which no reaction liquid, or only a very small amount of reaction liquid, is located temporarily because of the centrifugal acceleration. The method according to the invention for recording of the process parameters of reaction liquids will be explained in more detail in the following text, on the basis of two different variants, by means of an apparatus as shown in figure 2. Figure 4 illustrates one method for successive recording of the values of process parameters in in each case only one microreactor, with the sensor optics device (17) subsequently being moved by means of the positioning unit (7) . The sensor optics device (17) is aligned under one microreactor in such a manner that the electromagnetic radiation, at a wavelength between 200 nm - 25 µm, is introduced exclusively into this microreactor in the form of the light beam (18) for recording of individual measured values during the reaction. If a cylindrical microreactor is used having a circular-cylindrical bottom (10), the shaking diameter (27) of the rotation shaker (5, 6) about the eccentric axis (21) (see Figure 4) is chosen such that the light beam (18) from the sensor optics device (17) strikes only the bottom (10) of one of the microreactors. For this purpose, the shaking diameter (27) must be chosen to be less than or equal to the diameter (28) of the bottom (10). In the case of reactions which take place quickly, it is possible in order to speed up the recording process to combine groups of microreactors in a microreactor array, with the process parameters of the individual groups being recorded successively, but with the process parameters in the microreactors in one group being recorded at the same time by one sensor optics device (17) for each microreactor in the group. The sensor optics devices for simultaneous recording within one group are arranged on the positioning unit (7). After simultaneous recording of the process parameters in the microreactors in one group, the sensor optics devices are moved to the next group. The sensor optics devices for one group are aligned under the microreactors in that group in such a manner that the electromagnetic radiation of each sensor optics device for the recording of individual measured values is introduced exclusively into that microreactor under which the sensor optics device is currently located. In order to avoid signal flooding of the sensor, for example of a photodetector, by light reflections of the excitation light on the bottom (10), each sensor optics device is aligned with respect to each microreactor in such a way that none of the electromagnetic radiation which is reflected from the walls, in particular the bottom (10) of the microreactor, strikes the sensor. For this purpose, the optical waveguide end which is used as the sensor optics device (17) is positioned at an acute angle (22) with respect to a perpendicular to the bottom (10) of the microreactor. Different optimal positioning angles (22) are used, depending on the numerical aperture of the optical waveguide, preferably between 25° and 40°. Instead of successive recording of the process parameters it is, however, also possible to record the process parameters in all of the microreactors at the same time by one sensor optics device for each microreactor. The sensor optics devices are aligned under the microreactors in such a manner that the electromagnetic radiation (200 nm - 25 µm) is introduced exclusively into that microreactor which is associated with that sensor optics device, in the form of the light beam (18) for recording of the measured values during the reaction. Figure 5 illustrates one method for successive recording of the process parameters in four microreactors which are arranged adjacent to one another. The process parameters of the microreactors are recorded by the stationary sensor optics device (17) successively, on the basis of the relative circular movement with respect to this sensor optics device (17). The electromagnetic radiation of the sensor optics device (17) is introduced, during the recording of the process parameters in one of the four microreactors, exclusively into this microreactor, The radiation which emerges from the reaction liquid strikes exclusively that sensor of the sensor optics device (17) . Simultaneous injection into two adjacent microreactors is avoided because of the arrangement of the sensor optics device (17) with respect to the bottom (10) of the microreactors. The sensor optics device (17) is aligned with one of the four microreactors. If the shaking diameter is chosen to be equal to the diagonal distance between the center points of the microreactors, with the microreactors being distributed as illustrated on the tray (4) , then the 4 microreactors circle successively over the light beam (18) of the sensor optics device (17) during one revolution. The microreactors are moved over the sensor optics device (17) in the sequence I-IV, with the light beam (10), which is in a fixed position during the recording process, describing a circular line (23) . The sensor signals which are caused by the individual microreactors are recorded by means of the computer (8) , which is not illustrated here. In the course of the evaluation of the sensor signals, those sensor signals and microreactors from which the electromagnetic radiation which is responsible for the sensor signals originates are associated with one another. The microreactor position information that is required for the association process can be recorded, for example, by means of a position sensor (for example a light barrier, an acceleration sensor or a Hall sensor) which is arranged on the rotation shaker (5, 6) . If the microtiterplate (1) has more than four microreactors as illustrated in figure 5, the sensor optics device (17) is moved to a next adjacent group of microreactors (which are arranged such that they match one another) with four microreactors, after recording of the process parameters for the first group of four microreactors by the positioning unit (7). A positioning angle of the sensor optics device (17) at an angle (22) of between 25° and 40° with respect to bottom (10) of the microreactors has also been found to be advantageous for this method. However, the method which has been explained in principle with reference to figure 5 can also be used to record the process parameters of a greater number (>4) of microreactors per group. The shaking diameter (27) must then be chosen such that the microreactors describe a circle around the fixed-position sensor optics device, in which more than four microreactors are recorded successively during one revolution. In order to speed up the recording process, it is possible in a method as shown in figure 5 or figure 6C for the process parameters of a plurality of groups of microreactors arranged adjacent to one another to each be recorded in parallel by one sensor optics device for each group. According to the invention, the reaction liquids in the microreactors may have at least one chemical sensor material, which is preferably applied to at least one inner wall of the microreactor, for example to the bottom (10) . Chemical sensors such as these are, for example, fluorescent dyes which act as indicators of process parameters such as pH, T, pO2 and pCO2. In dissolved form, the fluorescent dyes can be used to determine the process parameters using the apparatuses and method as described with reference to figures 1-5. If the fluorescent dyes are immobilized on the inner walls as sensitive layers (38, 39), a specific alignment of the sensor optics device (17) is required, as is illustrated by way of example in figure 6A-B for the measurement method shown in figure 4. In this case, a plurality of sensitive layers (38, 39) can be arranged on the bottom (10) of the microreactor in such a way that different process parameters can be recorded. A cutout in the transparent bottom (10) is left free in order that the light beam (18) can also enter the liquid volume without any impediment. As shown in figure 6A, the alignment of the sensor optics device (17) and the shaking diameter (27) is to be matched such that the light beam (18) always strikes the sensitive layer (38, 39) or the cutout between them during one revolution of the tray (4) (shaking diameter (27) (N)+1) - length of a sensitive layer (41)]). As is illustrated in 41, the length of a sensitive layer in this case means the longest geometric dimension of a sensitive layer (38, 39). With regard to the recording of the measured values, it follows from this that the sensor optics device (17) is moved to different positions 35, 36 and 37 underneath the bottom (10) of the microreactor with the aid of the XY positioning unit (7) , in order to record different process parameters. Position 35 is used to record the natural fluorescence or the stray light of the reaction liquid, while positions 36, 37 are used to record the emission from the various sensitive layers 38, 39. In the variant shown in figure 6B, the various process parameters are recorded in a microreactor without changing the position of the sensor optics device between two recording processes. In this case, the light beam (18) successively strikes the sensitive layers (38, 39), which are immobilized on the bottom, on a circular path. A part of the bottom surface is free in order to allow direct injection into the reaction liquid, as is required for example for recording of specific process parameters, such as the stray light intensity or natural fluorescence of the reaction liquid. The various sensor signals are associated throughout the rotation of the microreactor by means of a position sensor, which is not illustrated. In the variant shown in figure 6C, the sensitive layers (38, 39) are fitted in the microreactors such that a plurality of microreactors are recorded successively by the light beam (18) during one revolution of the tray. The surface (40) which is covered by the sensor optics device (17) records four microreactors and different process parameters, which must be associated with the individual microreactors after the recording of the signals. In this case, this is once again done by means of a position sensor. If the microreactor array comprises more than the four illustrated microreactors, the sensor optics device is moved by the positioning unit (7) to further groups of microreactors, which are each arranged adjacent to one another and each have four microreactors, after recording the process parameters for the first group of four microreactors. When microreactors are used to cultivate microorganisms, they preferably have an upper opening which can be covered with a gas-permeable cover during operation, for example in the form a self-adhesive membrane. This membrane allows monoseptic operation of the microreactors. The reaction which takes place in the reaction liquid is supplied with the necessary gaseous reaction component, but is not impeded by gaseous reaction products. An one example of the recording of process parameters which have been determined using the method according to the invention, figure 7 shows measurement curves for a Hansenula polymorphs wt in 2xYP-medium (Yeast-Peptone) culture with 10g/L glycerin at a temperature of 27°C. The process parameters that were observed were the relative stray light intensity (at 620 nm) and the relative NADH fluorescence (excited at 340 nm, emission at 460 nm) of the culture throughout the duration of the reaction. The individual process parameters were recorded using a Y optical waveguide with a positioning angle of 30° and at a distance of 1 mm from the microtiterplate bottom. The shaking frequency in this case corresponded to 995 rpm, and the shaking diameter (27) was 3 mm. The reaction was carried out using a conventional 48-well microtiterplate (Greiner Bio-one, Frickenhausen, Part No.: 677 102) and a filling volume of 600 µl. A gas-permeable adhesive film was used as a cover (Abgene, Hamburg, Part No.: AB-0718), in order to cover the 48-well microtiterplate. We Claim: 1. A method for recording process parameters of reaction liquids in a plurality of microreactors (10, 19) of a microtitre plate, which are continuously shaken at least until the end of the reaction in all of the microreactors by means of a rotation shaker, wherein (a) the process parameters in the microreactors are recorded during the reaction whilst shaking continuously, (b) the process parameters are recorded with the aid of a sensor optics device (17) comprising a sensor (15) and a radiation source (12), (c) the process parameters in the individual microreactors (10, 19) are recorded successively by a sensor optics device (17) disposed on a positioning unit (7), (d) the electromagnetic radiation from the radiation source (12) is successively introduced into the reaction liquid of the microreactors of the microtitre plate and the sensor (15) associated with the radiation source records the electromagnetic radiation emanating from the reaction liquid of the microreactor, (e) after recording the process parameters in one of the microreactors, the sensor optics device (17) is moved by the positioning unit (7) to another microreactor, characterised in (f) that the sensor optics device (17) placed and aligned under one of the microreactors is not moved at least during the recording of the process parameters so that shaken microreactors (10,19) move relative to the sensor optics device (17), (g) wherein the shaking diameter of the rotation shaker is selected to be smaller than the diameter of the bottom (10) of the microreactors so that during the recording of the process parameters in one of the microreactors the electromagnetic radiation of the sensor optics device (17) is introduced exclusively into this microreactor and the electromagnetic radiation emanating from the said microreactor following the introduced electromagnetic radiation is exclusively recorded by the sensor (15) of the sensor optics device (17) associated with the introducing radiation source (12). 2. The method for recording process parameters of reaction liquids in a plurality of microreactors (10, 19) of a microtitre plate, which are continuously shaken at least until the end of the reaction in all of the microreactors by means of a rotation shaker, wherein (a) the process parameters in the microreactors are recorded during the reaction whilst shaking continuously, (b) the process parameters are recorded with the aid of a plurality of sensor optics devices (17) each comprising a sensor (15) and a radiation source (12), (c) the process parameters in the plurality of microreactors (10, 19) are recorded simultaneously by a plurality of sensor optics devices (17) and (d) the electromagnetic radiation from each radiation source (12) is successively introduced into me reaction liquid of the microreactors of the microtitre plate and the sensor (15) associated with the radiation source records the electromagnetic radiation emanating from the reaction liquid of the microreactor, characterised in (e) that each sensor optics device (17) placed and aligned under one of the microreactors (10, 19) is not moved at least during the recording of the process parameters so that shaken microreactors (10, 19) move relative to each sensor optics device, (f) wherein the shaking diameter of the rotation shaker is selected to be smaller than the diameter of the bottom (10) of the microreactors so that during the recording of the process parameters in one of the microreactors the electromagnetic radiation of each sensor optics device (17) is introduced exclusively into the microreactor below which the sensor optics device is specifically located and the electromagnetic radiation emanating from each microreactor following the introduced electromagnetic radiation is exclusively recorded by the sensor (15) of the sensor optics device (17) associated with the introducing radiation source (12). 3. The method as claimed in claim 2, wherein a microreactor array (1) comprises a plurality of groups of microreactors (10, 19), wherein the process parameters of the groups are recorded successively whilst the process parameters in the microreactors of one group are recorded simultaneously by a sensor optics device (17) for each microreactor in the group and after the simultaneous recording of the process parameters in the microreactors of one group, the sensor optics devices (17) disposed on a positioning unit (7) are moved by the positioning unit to another group. 4. The method for recording process parameters of reaction liquids in a plurality of microreactors (10, 19) of a microtitre plate, which are continuously shaken at least until the end of the reaction in all of the microreactors by means of a rotation shaker, wherein (a) the process parameters in the microreactors are recorded during the reaction whilst shaking continuously with the aid of at least one sensor optics device (17) each comprising a sensor (15) and a radiation source (12), (b) the electromagnetic radiation from the radiation source (12) is successively introduced into the reaction liquid of the microreactors and the sensor (15) associated with the radiation source records the electromagnetic radiation emanating from the reaction liquid of the microreactor, characterised in that (c) each sensor optics device (17) placed and aligned under one of the microreactors is not moved at least during the recording of the process parameters so that the shaken microreactors (10,19) move relative to each sensor optics device, (d) the process parameters of at least one group of adjacently disposed microreactors are successively recorded by this sensor optics system as a result of their relative movement (23) to the sensor optics device (17) which is not moved, (e) wherein the shaking diameter (27) of the rotation shaker is tuned so that the microreactors (10, 19) of each group successively circle over the sensor optics device (17) during a revolution of the rotation shaker and wherein the electromagnetic radiation emanating from each microreactor following the introduced electromagnetic radiation is exclusively recorded by the sensor (15) of the sensor optics device (17) associated with the introducing radiation source (12), (f) within the scope of an evaluation of the sensor signals, an assignment is made between the sensor signals and the microreactors (10, 19) from which the electromagnetic radiation emanates and (g) after recording the process parameters of one group (I-IV) of microreactors, the sensor optics device (17) is optionally moved by a positioning unit (7) to further groups (I-IV) of adjacently disposed microreactors (10,19). 5. The method as claimed in claim 4, wherein the process parameters of a plurality of groups of adjacently disposed microreactors are recorded in parallel by means of respectively one sensor optics device for each group. 6. The method as claimed in any one of claims 1 to 5, wherein a flashlamp is used as the radiation source (12) of the sensor optics device (17), its pulse frequency being matched to the shaking movement (27) in such a manner that the light flashes are incident on the microreactor (10, 19) at at least four different points (32). 7. The method as claimed in any one of claims 1 to 5, wherein a flashlamp is used as the radiation source (12) of the sensor optics device (17), its pulse frequency being matched to the shaking movement (27) in such a manner that the light flashes are always incident on the microreactor (10, 19) at the same point. 8. The method as claimed in any one of claims 1 to 7, wherein the electromagnetic radiation of each radiation source (12) and/or the radiation emanating from the reaction liquid is guided via optical waveguides (2, 25). 9. The method as claimed in any one of claims 1 to 8, wherein each sensor optics device (17) is aligned with respect to each microreactor (10, 19) in such a manner that none of the electromagnetic radiation which is reflected from the walls of the microreactor is incident on the sensor (15). 10. The method as claimed in any one of claims 1 to 9, wherein the reaction liquids in the microreactors have at least one chemical sensor material. 11. The method as claimed in claim 10, wherein the chemical sensor material (38, 39) is applied to at least one inner surface (10) of the microreactor. 12. An apparatus for carrying out the method as claimed in any one of claims 1 to 3, comprising: a microreactor platform (4) connected to a shaking device (5, 6), having at least two microreactors (10, 19) which are at least partially transparent to electromagnetic radiation, at least one sensor optics device (17) decoupled from the shaking movement (27) of the microreactor platform (4), each comprising a sensor (15) and a radiation source (12), wherein the radiation source (12) introduces electromagnetic radiation into the reaction liquid of the microreactors and the sensor (15) which is associated with said radiation source detects electromagnetic radiation emanating from said reaction liquid of said microreactors, as well as a data processing unit (8, 9) for recording and evaluating the sensor signals, characterised in that each sensor optics device (17) is fastened to a positioning unit (7) which is mounted in a fixed position with respect to the microreactor platform (4), said planar microreactor platform (4) is at least partially transparent to electromagnetic radiation in the area of bottom surfaces (10) of said microreactors (10, 19), each sensor optics device (17) can be moved by means of the positioning unit (7) under the bottom surfaces (10) of the microreactors, the shaking device (5, 6) is a rotation shaker, whose shaking diameter is smaller than the diameter (28) of the bottom of the microreactors (10,19) and the planar microreactor platform (4) is a tray having a receptacle into which a microtitre plate (1) is inserted. 13. The apparatus for carrying out the method as claimed in any one of claims 4 or 5, comprising: a microreactor platform (4) connected to a shaking device (5, 6), having at least two microreactors (10, 19) which are at least partially transparent to electromagnetic radiation, at least one sensor optics device (17) decoupled from the shaking movement (27) of the microreactor platform (4), each comprising a sensor (15) and a radiation source (12), wherein the radiation source (12) introduces electromagnetic radiation into the reaction liquid of the microreactors and the sensor (15) which is associated with said radiation source detects electromagnetic radiation emanating from said reaction liquid of said microreactors, as well as a data processing unit (8, 9) for recording and evaluating the sensor signals, characterised in that each sensor optics device (17) is fastened to a positioning unit (7) which is mounted in a fixed position with respect to the microreactor platform (4), said planar microreactor platform (4) is at least partially transparent to electromagnetic radiation in the area of bottom surfaces (10) of said microreactors (10, 19), each sensor optics device (17) can be moved by means of the positioning unit (7) under the bottom surfaces (10) of the microreactors, the shaking device (5, 6) is a rotation shaker, whose shaking diameter is tuned such that a plurality of microreactors (10, 19) successively circle over the sensor optics device (17) during a revolution of the rotation shaker, - the planar microreactor platform (4) is a tray having a receptacle into which a microtitre plate (1) is inserted. ABSTRACT METHOD AND DEVICE FOR RECORDING PROCESS PARAMETERS OF REACTION FLUIDS IN SEVERAL AGITATED MICROREACTORS The invention relates to a method for recording process parameters of reaction liquids in a plurality of microreactors (10, 19) of a microtitre plate, which are continuously shaken at least until the end of the reaction in all of the microreactors by means of a rotation shaker, wherein (a) the process parameters in the microreactors are recorded during the reaction whilst shaking continuously, (b) the process parameters are recorded with the aid of a sensor optics device (17) comprising a sensor (15) and a radiation source (12), (c) the process parameters in the individual microreactors (10, 19) are recorded successively by a sensor optics device (17) disposed on a positioning unit (7), (d) the electromagnetic radiation from the radiation source (12) is successively introduced into the reaction liquid of the microreactors of the microtitre plate and the sensor (15) associated with the radiation source records the electromagnetic radiation emanating from the reaction liquid of the microreactor, (e) after recording the process parameters in one of the microreactors, the sensor optics device (17) is moved by the positioning unit (7) to another microreactor, characterised in (f) that the sensor optics device (17) aligned under one of the microreactors is not moved at least during the recording of the process parameters so that shaken microreactors (10, 19) move relative to the sensor optics device (17), (g) wherein the shaking diameter of the rotation shaker is selected to be smaller than the diameter of the bottom (10) of the microreactors so that during the recording of the process parameters in one of the microreactors the electromagnetic radiation of the sensor optics device (17) is introduced exclusively into this microreactor and the electromagnetic radiation emanating from each microreactor following the introduced electromagnetic radiation is exclusively recorded by the sensor (15) of the sensor optics device (17) associated with the introducing radiation source (12). The invention also relates to an apparatus for carrying out the method as claimed.

Documents:

02796-kolnp-2006-abstract.pdf

02796-kolnp-2006-assignment.pdf

02796-kolnp-2006-claims.pdf

02796-kolnp-2006-correspondence others.pdf

02796-kolnp-2006-correspondence-1.1.pdf

02796-kolnp-2006-description (complete).pdf

02796-kolnp-2006-drawings.pdf

02796-kolnp-2006-form-1.pdf

02796-kolnp-2006-form-3-1.1.pdf

02796-kolnp-2006-form-3.pdf

02796-kolnp-2006-form-5.pdf

02796-kolnp-2006-g.p.a.pdf

02796-kolnp-2006-international publication.pdf

02796-kolnp-2006-international search authority report.pdf

02796-kolnp-2006-other document.pdf

02796-kolnp-2006-pct other.pdf

2796-KOLNP-2006-(10-04-2012)-CORRESPONDENCE.pdf

2796-KOLNP-2006-(10-04-2012)-OTHERS.pdf

2796-KOLNP-2006-(16-08-2012)-ABSTRACT.pdf

2796-KOLNP-2006-(16-08-2012)-AMANDED CLAIMS.pdf

2796-KOLNP-2006-(16-08-2012)-AMANDED PAGES OF SPECIFICATION.pdf

2796-KOLNP-2006-(16-08-2012)-ANNEXURE TO FORM 3.pdf

2796-KOLNP-2006-(16-08-2012)-DESCRIPTION (COMPLETE).pdf

2796-KOLNP-2006-(16-08-2012)-DRAWINGS.pdf

2796-KOLNP-2006-(16-08-2012)-EXAMINATION REPORT REPLY RECEIVED.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-1.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-13-1.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-13-2.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-13.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-2.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-3.pdf

2796-KOLNP-2006-(16-08-2012)-FORM-5.pdf

2796-KOLNP-2006-(16-08-2012)-OTHERS.pdf

2796-KOLNP-2006-(16-08-2012)-PA-CERTIFIED COPIES.pdf

2796-KOLNP-2006-(16-08-2012)-PETITION UNDER RULE 137.pdf

2796-KOLNP-2006-(22-08-2013)-AMANDED PAGES OF SPECIFICATION.pdf

2796-KOLNP-2006-(22-08-2013)-CLAIMS.pdf

2796-KOLNP-2006-(22-08-2013)-CORRESPONDENCE.pdf

2796-KOLNP-2006-(22-08-2013)-OTHERS.pdf

2796-KOLNP-2006-(22-08-2013)-PA.pdf

2796-KOLNP-2006-(25-02-2013)-CORRESPONDENCE.pdf

2796-KOLNP-2006-(25-02-2013)-FORM 3.pdf

2796-KOLNP-2006-(29-08-2012)-CORRESPONDENCE.pdf

2796-KOLNP-2006-(29-08-2012)-FORM-3.pdf

2796-KOLNP-2006-(29-08-2012)-OTHERS.pdf

2796-KOLNP-2006-ASSIGNMENT.pdf

2796-KOLNP-2006-CANCELLED PAGES.pdf

2796-KOLNP-2006-CORRESPONDENCE.1.2.pdf

2796-KOLNP-2006-EXAMINATION REPORT.pdf

2796-KOLNP-2006-FORM 13.1.2.pdf

2796-kolnp-2006-form 18.pdf

2796-KOLNP-2006-GPA.pdf

2796-KOLNP-2006-GRANTED-ABSTRACT.pdf

2796-KOLNP-2006-GRANTED-CLAIMS.pdf

2796-KOLNP-2006-GRANTED-DESCRIPTION (COMPLETE).pdf

2796-KOLNP-2006-GRANTED-DRAWINGS.pdf

2796-KOLNP-2006-GRANTED-FORM 1.pdf

2796-KOLNP-2006-GRANTED-FORM 2.pdf

2796-KOLNP-2006-GRANTED-FORM 3.pdf

2796-KOLNP-2006-GRANTED-FORM 5.pdf

2796-KOLNP-2006-GRANTED-SPECIFICATION-COMPLETE.pdf

2796-KOLNP-2006-INTERNATIONAL PUBLICATION.pdf

2796-KOLNP-2006-INTERNATIONAL SEARCH REPORT & OTHERS.pdf

2796-KOLNP-2006-OTHERS.pdf

2796-KOLNP-2006-TRANSLATED COPY OF PRIORITY DOCUMENT.pdf

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