EP2044420A2

EP2044420A2 - Analytical system comprising an arrangement for temporally variable spatial light modulation and detection method implemented thereby

Info

Publication number: EP2044420A2
Application number: EP07805169A
Authority: EP
Inventors: Max Wiki
Original assignee: Individual
Current assignee: Individual
Priority date: 2006-07-17
Filing date: 2007-07-16
Publication date: 2009-04-08
Also published as: WO2008010182A2; WO2008010182A3; US8213017B2; US20100014088A1

Abstract

The invention relates to an analytical system and a method carried out thereby for producing and measuring optical signals and/or the variations thereof from measuring regions which are arranged in a one-dimensional or two-dimensional array on an essentially optically transparent sensor platform (2). Said system comprises at least: an optical system provided with a lighting system (1) for illuminating measuring regions on the sensor platform (2), and a detection system (3) provided with at least one detection unit (11) for detecting signals from the measuring regions on the sensor platform, in the direction of the transmission or reflection of the illumination light in a spectral region comprising the spectral region of the illumination light; and a sensor platform (2) that can be introduced into the optical system and comprises measuring regions arranged thereon in a one-dimensional or two-dimensional array. The invention is characterised in that the illumination system (1) comprises an arrangement for rapidly variable spatial light modulation, described as an 'SLM' (8.4), by which means, in the operating state, illumination patterns having freely selectable and rapidly variable dimensions which can be determined by the adjustments of the SLM (8.4) can be generated on the sensor platform (2), from an illumination light entering into the SLM (8.4), with essentially homogeneous intensity distribution in the cross-section of the illumination light, perpendicularly to the propagation direction thereof. The invention also relates to the use of the claimed analytical system and to the claimed method implemented thereby.

Description

Analytical system with an arrangement for temporally variable spatial light modulation and thus executable detection method

Technical area

The present invention relates to an analytical system and method to be carried out therewith for the generation and measurement of optical signals from measuring areas, which are arranged in a one- or two-dimensional array on a solid support as a measuring platform, which is referred to below as a sensor platform. Such an analytical system and method is useful, for example, for analyzing samples for biological, biochemical or synthetically produced substances contained therein as analytes. In particular, the invention relates to such a system and method which is suitable for simultaneously examining a single sample for a plurality of analytes and / or a plurality of samples for one or more analytes on a common sensor platform. In this case, the analytical system according to the invention is suitable for refractive measurement methods.

State of the art

For the simultaneous determination of a large number of analytes, numerous embodiments of so-called "microarrays" have become known in recent years in which a multiplicity of different biological or biochemical or synthetic recognition elements, for the specific recognition and binding of the analytes to be detected, in discrete measurement ranges, also as " Spots ", are immobilized. In many cases, the analyte detection is based on the detection of optical signals, for example of luminescence or luminescence changes luminescent molecules which are linked to the analyte itself or one of its binding partners which bind to the respective recognition elements in one or more binding assay steps in the measurement regions. Such microarrays, in which the analytes to be detected from a sample brought together with the array, bind to their recognition elements immobilized in the spots, after they have been recognized by these and "captured", are also referred to as "capture arrays".

In addition to these relatively widespread "capture arrays", so-called "reverse arrays" or "reverse phase arrays" have become known in recent years, in which the samples themselves, for example, untreated or only a few sample preparation steps subjected cell lysates, in discrete measurement ranges are immobilized in a microarray and the microarray then the recognition elements, optionally linked to luminescent or fluorescent Labein be supplied for signal generation in luminescence-based detection method. Such arrays have been described, for example, in international patent applications WO 2004/023142 and WO 2004/023143. The analytical system and method of the invention may be used in combination with both types of arrays, i. both "capture arrays" and "reverse arrays" are used.

For the detection of the analyte binding in different measuring ranges, a spatially resolved detection of the of the

Measuring ranges of outgoing light advantageous, the spatial resolution of the detection step, according to the arrangement of the measuring ranges, should be one or two-dimensional. This is usually, for the simultaneous detection of the signals from a variety of measuring ranges, by the use of appropriate spatially resolving detectors (eg Line detectors for one-dimensional spatial resolution or cameras for two-dimensional spatial resolution) achieved.

Based on simple glass or microscope slides and fluorescence detection for analyte detection, arrays of very high feature density (i.e., number of discrete measurement areas per unit area) are known. For example, U.S. Patent No. 5,445,934 describes and claims arrays of oligonucleotides having a density of greater than 1000 features per square centimeter. The excitation and readout of such arrays is based on classical optical arrangements and methods. The entire array can be illuminated simultaneously with an expanded excitation light beam, but this leads to a relatively low sensitivity, since the scattered light component is relatively large and scattered light or background fluorescent light is generated from the glass substrate even in those areas in which no binding of the analyte immobilized recognition elements are located. In order to limit the excitation and detection to the areas of the immobilized features and to suppress light generation in the neighboring areas, often confocal measuring arrangements are used and the different features are sequentially read out by means of "scanning". However, this can result in a greater amount of time spent reading a large array and a relatively complex optical design.

There are a number of different types of sensor platforms and thus feasible detection method known, which differ for example in the measuring principle used and can be divided into different categories accordingly. For example, a distinction can be made between luminescence-based detection methods, for example based on fluorescence or phosphorescence detection after optical excitation of fluorescent or phosphorescent groups used in the detection step or Molecules, and refractive measurement methods by means of which changes in the refractive index are detected on the surface of a sensor platform. The refractive measurement methods are also referred to as "marker-free"("label-free") methods, which have the advantage that they do not "label" or link the compounds to be detected with a molecular group to be used specifically for the detection step In the case of refractive measurement methods, the signals to be detected and their changes may be due, for example, to near field effects, which lead to a surface-bound signal amplification (see below).

Evanescent field sensor platforms are often used to improve sensitivity, i. Such measuring platforms, on whose surface facing the sample to be examined an evanescent field, also referred to as transversely attenuated field, can be generated, which limits the interaction of the analyte and / or the sample excitation or measurement light on the penetration depth of this evanescent field in an adjacent Medium on the order of a fraction of the wavelength of the excitation or measuring light used allows. Thus, a spatially highly selective excitation of molecules or interaction with molecules within the penetration depth of this field in the adjacent media is possible, while at the same time interference signals from areas beyond this penetration depth can be avoided.

An evanescent field is produced, for example, by total reflection of a light beam propagating in a medium (eg, a prism or a self-sustaining optical waveguide such as a glass plate) having a higher refractive index than that of the surrounding medium at the interface of this higher refractive index medium to the low refractive medium. In such a configuration, the evanescent field is in each case discrete locations of total reflections produced at the interfaces to the low-refractive medium.

As a further development of optical waveguides are optical thin-film waveguides, in particular planar optical

Thin-film waveguides are known, which are in the simplest case a three-layer system: carrier material (often referred to as "substrate"), waveguiding layer, superstrate (or sample to be examined), wherein the waveguiding layer has the highest refractive index The strength of the evanescent field is very much dependent on the thickness of the waveguiding layer itself and on the ratio of the refractive indices of the waveguiding layer and the surrounding media, for thin waveguides, ie layer thicknesses of the same or lower thickness as the wavelength to be guided, discrete modes of the guided light can be discriminated.

Evanescent fields can also be generated by means of so-called "resonant lattice structures." Resonant lattice structures, like planar optical waveguides, have a structuring (stratification) with materials of different refractive indices in the direction perpendicular to the plane of the substantially planar substrate, with at least one of them on the substrate In addition to (in the plane parallel to that of the substrate unstructured) optical waveguides resonant lattice structures have a one-dimensional or two-dimensional, mostly Periodic structuring of the (effective) refractive index, generated using the same or different materials, in the plane of the at least one higher refractive index layer parallel to the substrate plane. In addition, a resonant grid structure may have a structuring with materials of different refractive indices, as in the direction perpendicular to the substrate plane, also in the plane parallel thereto. Resonant grating structures are known from the literature and are described, for example, in US Pat. No. 5,598,300 and in patent application US 2004/0130787.

As a further embodiment of evanescent field sensor platforms, so-called "resonant mirrors" may be considered which essentially comprise a three-layer system: a high refractive prism with a lower refractive index layer deposited thereon beneath another high refractive index layer thereon , Under operating conditions, light incident on the prism is totally reflected at its surface. The resulting in this total reflection at the surface of the prism evanescent field extends into the layer with low refractive index, and due to the small thickness of the low-refractive layer, it also extends into the high refractive second layer. The second high-index layer acts like a waveguide. The effective refractive index of this waveguide, and thus the propagation speed of the light in this waveguide, is determined by the optical parameters of the layer system (essentially refractive indices and layer thicknesses) and a coating optionally located on the surface of the evanescent field sensor platform (eg with an adhesion-promoting layer or layer) with specific recognition elements for analyte detection) and of the surrounding medium. When the propagation velocity of the evanescent field of the prism coincides with the propagation velocity of the light in the waveguide, part of the evanescent field of the prism becomes the waveguide coupled. The propagation velocity of the evanescent field of the primate can be influenced, for example, by changing the angle of incidence or by changing the wavelength. The guided in the waveguide light again evanescent fields arise on both sides of the waveguide, the prism-side evanescent field of the waveguide in turn extends into the low-refractive layer and beyond into the high refractive prism. Analogous to the coupling into the waveguide, light guided in the waveguide is decoupled again into the prism and can be directed from there to a detection unit. The processes of light coupling and decoupling bound to the prism and the second high refractive index layer occur simultaneously, whereby the light directed from the prism to the detection unit has a resonant characteristic. This "resonant mirror" technology is more fully described, for example, in US Pat. No. 5,255,075, and in PE Buckle et al., "The resonant mirror: a novel optical sensor for direct sensing of biomolecular interactions, Part II: Application," Biosensors & Bioelectronics 8 (1993), 355-363.

The use of evanescent field sensor platforms is suitable to solve the problems of providing improved sensitivity, ie. H. lower detection limits for analyte detection, and to solve a higher accuracy. In addition, however, there is a need for the optimization of analytical systems and detection methods for the measurement of the largest possible number of signals from a plurality of discrete measurement ranges within the shortest possible time.

For example, imaging methods and analytical systems equipped therefor are known. Known from the prior art

Measuring arrangements with simultaneous more or less large area Illumination of a multiplicity of measuring ranges on a sensor platform, for example for recording luminescence or fluorescence signals from microarrays on a thin-film waveguide as sensor platform from the international application WO 02/21110 or signals of a large-area grating coupler sensor (likewise based on a thin-film waveguide; Explanation see later) from international application WO 01/88511. These arrangements make it possible to record signals from a plurality of measuring ranges within a relatively short measuring time determined by the duration of the illumination. However, these arrangements have the disadvantage that, as a result of the continuous large-area illumination of the sensor platform, which is not limited to the regions of the measuring ranges, in particular signals from the intermediate ranges between the measuring ranges lead to an increased fundamental signal, which is not caused by the presence of an analyte which results in a reduced signal-to-noise ratio due to the resulting increase in background signal level, and hence reduced sensitivity for the determination of a variety of analytes as compared to sensitivity for a single analyte determination in a single measurement range limited excitation and detection can lead to a sensor platform otherwise similar technical design. In addition, there is an increased risk of optical crosstalk of the signals from the various measuring ranges with continuous large-area illumination.

The risk of crosstalk of optical signals from different measuring ranges is circumvented by scanning arrangements, ie sequential illumination and signal detection by different discrete measuring arrangements. Especially in the case of large-sized sensor platforms, for example in the format of a standard microtiter plate, but extended overall times for the Signal recording and possibly complex mechanical positioning for necessary translations over long travels, coupled with very high demands on the positioning accuracy to expect, which can additionally lead to an adverse increase in system costs.

It is therefore an object of the present invention to provide an analytical system and a method to be carried out with which it is possible to examine a sample for a plurality of analytes contained therein and / or a plurality of samples for one or more analytes contained therein within a short time high sensitivity, as is known, for example, from evanescent field sensor platforms for single analyte determinations, and which still has to be improved by an increase in the signal-to-noise ratio, possibly accompanied by a reduced risk of optical crosstalk between signals from different measurement ranges ,

Brief description of the invention

According to the invention the above object is achieved by means of an analytical system for generating and measuring optical signals and / or their changes from measuring ranges, which are arranged in a one- or two-dimensional array on a substantially optically transparent sensor platform, at least comprising - an optical system with an illumination system for illuminating measuring areas on the sensor platform and a detection system having at least one detection unit for detecting signals from the measuring areas on the sensor platform, in the direction of the transmission or reflection of the Illuminating light in a spectral region which comprises the spectral region of the illumination light, and a sensor platform which can be inserted into the optical system and has measuring regions arranged thereon in a one- or two-dimensional array, characterized in that the illumination system is an arrangement which can be rapidly changed over time, referred to as "SLM" Spatial light modulation includes, with the operating state of an entering into this SLM illumination light, with substantially homogeneous intensity distribution in the cross section of the illumination light perpendicular to its propagation direction, illumination patterns freely selectable and quickly changeable, can be determined by the settings of the SLM geometry on the sensor platform can be generated.

By "rapidly variable" spatial light modulation and "rapid change" of the illumination pattern is to be understood that these changes take place with a frequency greater than 1 Hz, preferably greater than 1 kHz, more preferably greater than 1 MHz.

The freely adjustable geometry of the illumination pattern on the

Sensor platform makes it possible to selectively illuminate measuring areas in any arrangement and geometry on the sensor platform and to selectively shade areas between the measuring areas, so that the generation of interfering scattered or interfering signals from these neighboring areas can be avoided. It can do that

Illumination patterns are changed at high speed. This advantageously allows an extremely fast optical "scanning" of the entire surface of the sensor platform with limitation of the signal excitation to the measuring areas themselves and detection of the signals from a plurality of measuring ranges, while avoiding the aforementioned disadvantages of scanning arrangements and methods. Compared to the prior art, the analytical system according to the invention has further significant advantages:

- The mechanical tolerances of the receiving device of the analytical system for a sensor platform to be introduced can be relatively generously dimensioned, since variations in the dimensions or the positioning of a sensor platform to be inserted can be compensated by adjustments of the illumination pattern. This can further the requirements for the tolerances of the dimensions of

Sensor platform are kept generous, which reduces their production costs.

- Preferred embodiments of the analytical system according to the invention with a plurality of individually controllable individual elements of the SLM, which is preferably greater than the number of

Measuring ranges, allow a high spatial resolution of the signal generation and thus signal detection and various forms of sub-sampling.

- In the case of illumination of measuring ranges by several individual elements of the SLM, several measured values per measuring range can be generated simultaneously independently of each other.

- It is not necessary to adapt or even change or replace the optical elements of the analytical system in a change in the geometric arrangement of measurement areas on the sensor platform, but the illumination pattern can be adjusted accordingly.

- The use of diffractive optical elements, which often produce a rigid, unchangeable illumination pattern, can be dispensed with in the beam paths. In particular, their otherwise required change or exchange in case of changes the light source or the mentioned geometric characteristics of the sensor platform can be avoided.

- It can also broadband light sources are used, which is hardly possible in combination with diffractive optical elements in the illumination beam path.

The rapid variability of the generated illumination pattern allows its adaptation to dynamic processes (eg the growth or contraction of biological cells.

- Different measuring ranges can have any, different, independent form.

- Binning or similar functions can be used.

- It is not necessary to use additional masks in the beam paths.

- The total number of optical components of the system can be reduced, resulting in a reduction of system costs: Several diffractive optical elements and masks can be replaced by a single SLM.

A further subject of the present invention is a method for generating and measuring optical signals and / or their changes from measuring ranges, which are arranged in a one- or two-dimensional array on a substantially optically transparent sensor platform, using an analytical system according to one of hereinafter described embodiments, at least comprising

- An optical system with an illumination system for illumination of measurement areas on the sensor platform, with a designated as "SLM" arrangement for temporally fast variable spatial light modulation and with a detection system with at least one detection unit for detecting signals from the measurement areas on the sensor platform in the direction of the Transmission or reflection of the illumination light in a spectral range, which includes the spectral range of the illumination light, and

- An insertable into the optical system sensor platform arranged thereon in a one- or two-dimensional array

Measuring ranges, characterized in that in the operating state from an entering into this SLM illuminating light, with substantially homogeneous intensity distribution in the cross section of the illumination light perpendicular to its propagation direction, illumination patterns freely selectable and quickly changeable, can be determined by the settings of the SLM geometry on the sensor platform can be generated.

Particular preference is given to such an embodiment of the method according to the invention, which is characterized in that the optical signals and / or their changes are generated from measuring ranges by one or more binding or Adsorptionsereignisse between one or more analytes in one or more samples and specific recognition elements for said analytes in or on said measurement areas, wherein the samples and the recognition elements for the analytes to be detected in the samples are brought into contact with each other on the measurement areas, and that from these optical signals and / or their changes a simultaneous qualitative and / or quantitative detection a plurality of analytes in one or more samples and / or one or more analytes in a plurality of samples. Brief description of the figures

FIG. 1 schematically shows a first embodiment of an analytical system according to the invention, in which the illumination system 1 and the detection system 3 are arranged on the same side with respect to the sensor platform 2.

FIG. 2 shows a preferred embodiment of an analytical system according to the invention with a sensor platform 2 designed as a microtiter plate, a beam splitter 9.1 and approximately vertical telecentric illumination of the sensor platform 2, and detection of the light emanating from the sensor platform in the direction of reflection of the illumination light.

FIG. 3 shows an embodiment of an analytical system according to the invention similar to FIG. 2, except for the beam splitter with respect to the electro-optical components used, but with illumination of the illumination light and detection of the light emanating from the sensor platform 2 at a deviating from the perpendicular, i. oblique angle.

FIG. 4 schematically shows an embodiment of an analytical system according to the invention, in which the illumination and the detection system are arranged on opposite sides with respect to the sensor platform and the detection takes place in the direction of the transmitted light.

FIG. 5 shows a preferred embodiment of an analytical system according to the invention with a sensor platform 2 designed as a microtiter plate, a beam splitter 9.1 and approximately vertical illumination of the sensor platform 2 and detection of the sensor platform outgoing light in the direction of the reflection of the illumination light by means of a spectrometer 11.1 as a light analysis unit.

FIG. 6 shows a preferred embodiment of an analytical system according to the invention with a microtiter plate

Sensor platform 2, a beam splitter 9.1 and approximately vertical illumination of the sensor platform 2 and detection of the emanating from the sensor platform light in the direction of reflection of the illumination light for different polarizations using two spectrometers 11.1 as light analysis units.

FIG. 7 shows an embodiment of an analytical system according to the invention of similar construction to that of FIG. 2 with a sensor platform 2 which is designed as a prism for generating total internal reflection or as a "resonant mirror".

FIG. 8 shows an embodiment of an analytical system according to the invention similar to that of FIG. 2, but with detection of the light emanating from the sensor platform 2 in the transmission direction.

9 shows, using the example of a microtiter plate as sensor platform, possible variations of the illumination pattern that can be generated on the sensor platform by means of a spatial light modulator SLM a) with illumination of a single well of the microtiter plate, marked "S" and surrounded by a hatched square b) simultaneous illumination (FIG. marked by hatched cross) of four surrounding wells (labeled "R").

FIG. 10 shows further illumination patterns with different geometries that can be generated on the sensor platform: a) Illumination of measurement areas in a hexagonal pattern; b) Illumination pattern off concentric circular rings; c) a lighting pattern with any irregular geometry, such as the targeted illumination of located on the sensor platform biological cells may correspond.

FIG. 11 shows the emission spectrum of a projection system used for examples 1 and 2 with three light-emitting diodes (LEDs) as light sources, with central emission wavelengths 460 nm (blue), 520 nm (green) and 627 nm (red), the spectral emission width of the LEDs each 20 nm to 40 nm.

FIG. 12 shows, for four differently set angles of incidence 6.3, 6.4, 6.5 and 6.6 of the illumination light on the sensor platform, the spectra of the TE-poled illumination light of the red LED from FIG. 11 taken in transmission.

FIG. 13 shows, for two differently set angles of incidence 6.1 and 6.2 of the illumination light on the sensor platform, the spectra of the TM-polarized illumination light of the red LED from FIG. 11 recorded in transmission.

FIG. 14 shows, for a single set angle of incidence of the illumination light on the sensor platform, the spectrum of the TM-polarized illumination light of the red LED from FIG. 11 recorded in the reflection direction.

Detailed description of the invention

For the purposes of the present invention, measuring ranges are to be defined in each case by the closed surface which is located on a sensor platform for Detection of a single measured variable is identified, that is used to generate a single signal value. By way of example, the sensor platform can be designed as a microtiter plate, with a multiplicity of cavities (wells) arranged in the form of a matrix, and a thin-film waveguide with gratings structured therein as a bottom.

For example, the bottom surface of a single well can then represent a measuring range on which the refractive index of a liquid introduced into the well is determined by a refractive measurement. However, the bottom surface of a single well may also serve to detect a plurality of readings for simultaneously detecting a plurality of analytes in a sample filled into a single well. In accordance with the abovementioned comprehensive definition, measurement areas should then be defined by the closed areas which contain immobilized compounds to be detected or specific binding partners immobilized there, for the detection of one or more analytes in one or more samples in a bioaffinity assay. These surfaces can have any geometry, for example the shape of circles, rectangles, triangles, ellipses, etc. Preferably, the measuring ranges are a plurality of measuring ranges. Particularly preferred are discrete, laterally or spatially separated measuring ranges.

Preferably, the sensor platform in a one- or two-dimensional arrangement comprises more than 50, more preferably more than 500, most preferably more than 50,000 measuring ranges.

There are a variety of known types or methods for applying or immobilizing the compounds or specific binding partners to be detected on a sensor platform. For example, this can be done by physical adsorption or by electrostatic interaction. The orientation of the immobilized compounds or the the specific binding partner is then generally random so that only a portion of them are accessible for binding with a binding partner to be delivered in a sample. In addition, there is a risk that with different composition of the sample containing the analyte or used in the detection method

Reagents a part of the immobilized compounds or binding partner is washed away. It may therefore be advantageous if, for the purpose of a more stable immobilization, an adhesion-promoting layer is applied to the surface of the sensor platform to be used for generating the signal. This adhesion-promoting layer should be optically transparent. Particularly in the case of evanescent field sensor platforms (see below), it is preferred that the adhesion-promoting layer does not project beyond the penetration depth of the evanescent field out of the high-index layer of the sensor platform into the medium adjacent thereto. Therefore, the primer layer should have a thickness of less than 200 nm, preferably less than 20 nm. It may comprise, for example, chemical compounds from the group of silanes, epoxides, functionalized, charged or polar polymers, and "self-assembled functionalized monomers."

Compounds or specific binding partners to be immobilized on a sensor platform, in particular also as specific recognition elements to be immobilized, and also the analytes to be detected are preferably selected from the group consisting of nucleic acids (for example DNA, RNA, oligonucleotides), nucleic acid analogs (eg PNA), Antibodies, aptamers, membrane-bound and isolated receptors whose ligands, antigens for antibodies, "histidine tag components", etc. is formed. It is also possible to immobilize entire biological cells or components thereof on a sensor platform. A single measurement area on a sensor platform may contain a single type of compound, for example, to detect a single analyte in a sample to be delivered. This is preferred in the case of "capture arrays" to be generated. However, a single measurement range can also contain a large number of different compounds. This is the case in the case of "reverse arrays" in which a single measuring range is formed in each case by the closed surface on the sensor platform, which occupies a sample immobilized there with a plurality of different compounds contained therein and to be detected as analytes.

As sensor platform insertable into the optical system of the analytical system according to the invention, various embodiments of sensor platforms are suitable. The sensor platform should be substantially optically transparent at least at the wavelength of the illumination light. By "essentially optically transparent" as a property of the sensor platform, which can consist of different layers, for example, it should be understood that light of the wavelength of the illumination light when propagating in the volume of the material or a layer material of the sensor platform over a distance of 5 mm, a weakening by absorption of not more than 80%, preferably not more than 50%, more preferably not more than 20%, propagation losses due to interfacial scattering, for example in light conduction in a sheet waveguide due to surface roughness or surface structuring such as a diffractive surface relief grating are not taken into account in this definition of "substantially optical transparency", even if they should even prevent the further propagation of light. By contrast, the aforementioned requirement for the "essentially optical transparency" of the sensor platform includes, for example, coatings with absorbing metals such as gold or silver, as they do For example, be used for sensor platforms for the generation of surface plasmon resonances and in which the propagation of a "coupled light" takes place only over a length of the order of a few hundred micrometers, and thus also such sensor platforms per se. The material of the sensor platform preferably comprises in a single-layer or multi-layer system a material from the group which is formed by substantially optically transparent glasses, plastics and ceramics, wherein layers of these materials may optionally be provided with additional coatings.

It is preferred that the side of the sensor platform on which the discrete measurement areas are created is substantially planar. In this case, "essentially planar" should be understood to mean that the radius of curvature of the surface of this side is greater than 10 cm in the region of the measuring ranges. For the purposes of this definition, for example, an embodiment of a sensor platform in the form of a microtiter plate, with the bottom surfaces of the wells as measuring regions, is also considered to be "substantially planar", since the above definition is fulfilled for the range of these measuring ranges. In addition, it is preferred that the ripple of the surface comprising the measuring areas of the

Sensor platform in the range of the measuring ranges is less than 0.2 mm. Furthermore, it is preferred that the roughness of the surface of the sensor platform comprising the measurement areas is less than 20 nm, more preferably less than 2 nm, most preferably less than 1 nm in the region of the measurement areas. The preference for surface roughness applies in particular to embodiments designed as evanescent field sensor platforms (see below) in order to minimize propagation losses of light due to scattering at the surface provided with the measurement areas. Intentionally performed surface structuring, such as diffractive surface relief gratings or other deliberately applied Surface structures should not be taken into account in this preference with regard to the "surface roughness" and their definition.

A preferred embodiment of the analytical system according to the invention is characterized in that the sensor platform is designed as an evanescent field sensor platform.

It is preferred that the sensor platform is selected from the

Group of prisms for generating total internal reflection, self-supporting optical waveguides, optical thin-film waveguides, thin-film waveguides with grids structured therein for light coupling and / or light outcoupling, resonant lattice structures and "resonant mirrors".

For embodiments of an analytical system according to the invention with a thin-film waveguide or a thin-film waveguide with gratings structured therein as evanescent field sensor platform, it is preferred that the carrier material of the waveguiding layer or the layer adjacent to the waveguiding layer in the direction of the side of the waveguiding layer which faces away from the measuring regions has the lowest possible Refractive index has, since this is favorably influenced the propagation of the evanescent field in the direction of the measuring regions on the opposite side of the waveguiding layer: The maximum guided in an optical waveguide intensity, based on the cross section of a slab waveguide, in the case of an asymmetric distribution of refractive indices the layers adjacent to the waveguiding layer are arranged asymmetrically in the direction of the neighboring layer with the higher refractive index.

Preferably, the refractive index of the carrier material or the layer adjacent to the waveguiding layer in the direction of the side of the waveguiding layer which faces away from the measuring regions is less than 1.6, particularly preferably less than 1.5, very particularly preferably less than 1.4. As materials with correspondingly low refractive indices, thermoplastic materials such as polyvinylidene fluoride, polymethylpentene, cycloolefin copolymers (COC) or cycloolefin polymers (COP) and mixtures of polyvinylidene fluorides and polymethylmethacrylates are suitable for this purpose, for example. Even fluoroplymers such as fluoroacrylate having a refractive index of less than 1.4 are known. When using the sensor platform for the analysis of liquid or aqueous samples, carrier materials with the lowest possible liquid or water absorption are advantageous. At the same time, it is advantageous if the waveguiding layer has the highest possible refractive index, preferably higher than 1.8, particularly preferably higher than 2.0. As materials of the waveguiding layer, metal oxides such as TiO ₂ , Ta ₂ O ₅ , Nb ₂ O ₅ , ZnO, HfO ₂ , ZrO ₂ , TiO ₂ -SiO ₂ , Al ₂ O ₃ , Si ₃ N ₄ , HfON, SiON are particularly good suitable. Such embodiments of sensor platforms which are suitable for an analytical system according to the invention are also described in US Application No. 2005/0025421 with FIGS. 13 (production of a "microplate" corresponding to FIG

Microtiter plate with thin-film waveguides with grids structured therein in the bottom of the wells of the microtiter plate), Figures 7, 8 and 9 (refractive index and distribution of mode intensity, ie intensity of the guided light), which are hereby incorporated in its entirety together with the corresponding parts of the description in this application become.

In refractive measurement methods using an evanescent field sensor platform, the change in the so-called effective refractive index due to changes in mass occupancy due to molecular adsorption or desorption on the sensor platform or Changes in the macroscopic refractive index of a surrounding medium used in the evanescent field of the sensor platform for an analyte detection. The effective refractive index is dependent on the macroscopic refractive index of the high-refractive, waveguiding layer and its thickness, the refractive indices and layer thicknesses of adjacent layers or media, and on the mode order of the light to be coupled via the grating into the waveguiding layer, the polarization (eg transversally electrical (eg. TE) or transverse magnetic (TM) polarization) as well as the wavelength of an irradiated illumination light.

The changes in the effective index of refraction, at a constant wavelength of incident illuminating light, result in changes in the resonance angle with respect to the surface normal of the sensor platform for light coupling in or out of a grating coupler sensor platform. These changes in resonance angles are determinable from the signals and their changes provided by the detection system (comprising one or more detectors) of the analytical system according to the invention. In the case of a constant angle of incidence of the illuminating light, changes in the resonance wavelength for the light coupling in or out of the corresponding detector signals, with a polychromatic spectrum of the illumination light being variable or over a certain spectral range (eg of the order of 1 to 20 nm), become apparent a grating coupler sensor platform determined. The state of the art in the use of grating coupler sensor platforms is described, for example, in US Pat. Nos. 4,815,169, 6,455,004 and 6,429,022, and in K. Tiefenthaler, W. Lukosz, "Sensitivity of grating couplers as integrated-optical chemical sensors". J. Opt. Soc. Am. B6 (1989) 209 et seq., W. Lukosz, PM Nellen, C. Stamm, P. Weiss, "Output grating couplers on planar waveguides as integrated, optical chemical sensors", Sensors and Actuators B1 (1990) 585 et seq. And T. Tamir, ST. Peng, "Analysis and design of grating couplers", Appl. Phys. 14 (1977) 235-254.

For the production of the coupling gratings, the person skilled in the art, for example, the use of injection molding, hot stamping, holographic exposure or so-called nanoimprint lithography and the techniques described in the patent application US 2003/0017581 and US Patent 6,873,764 known. Usually, the lattice structures are first produced as surface structures in a surface of the substrate, whereupon in a subsequent step, directly or on optional intermediate layers, at least one dielectric layer with preferably higher refractive index than the substrate and controlled layer thickness (of for example 50 nm to 500 nm) is applied and typically, the structure created on the substrate transmits to the surface of this dielectric layer in the deposition step. Resonant Mirror sensor platforms are fabricated by depositing dielectric layers of different refractive index and well-defined layer thickness on at least the wavelength of the illumination light in substantially optically transparent substrates.

In particular, for compatibility with established laboratory robots and the standard equipment of chemical and analytical laboratories, it is advantageous if the sensor platform has the basic dimensions of a microtiter plate according to SBS standard (about 85.5 mm x 127.8 mm) or is formed as part of a microtiter plate. This includes that the sensor platform itself can be a microtiter plate. Such arrangements are also disclosed in US 2005/0110989 (Figs. 2-6 and 14-16), US 5,738,825 (Figs. 1-3, 5-7) and US 6,018,388 (Figs. FIGS. 1-4), the named figures and associated descriptions of which are hereby incorporated in their entirety as part of the present invention

Registration will be introduced. If the sensor platform as a microtiter plate For example, it may comprise 96, 384 or 1536 sample containers or "wells" according to standard industrial manufacturing.

With regard to the arrangement for temporally variable spatial

Light modulation is preferred to be selected from the group of "Digital Mirror Devices" DMD, Liquid Crystal Displays LCD, "Liquid Cristal on Silicon Silicon" LCOS microdisplays, and mechanically movable masks with light transmissive and light blocking regions.

It is preferred that the arrangement for temporally variable spatial light modulation comprises a plurality of discretely for the transmission to the sensor platform or blockade of the illumination light switchable individual elements in a one- or two-dimensional arrangement. It is preferred that at least one of these individual elements corresponds to a measuring range. It is particularly preferred that a measuring range is illuminated by a plurality of these individual elements. By using a plurality of individual elements per measurement range, the illumination pattern can be better and more easily approximated to the exact shape of the measurement areas, such as rectangles, triangles, circles, hexagonal shapes, or any shapes, such as those found in cell measurement, and illumination of Areas that do not belong to the measurement areas, are largely avoided, resulting in an improved signal-to-noise ratio.

However, in each case one or more individual elements can be used to illuminate areas between the measuring areas, for example for generating background signals, calibration signals or referencing signals, which can be used to evaluate the signals from the measuring areas. It is particularly preferred that the arrangement for temporally variable spatial light modulation comprises a plurality discrete for the forwarding to the sensor platform or blockade of the illumination light switchable individual elements in a two-dimensional arrangement. Preferably, this involves more than 100 × 100 discretely switchable individual elements in a two-dimensional arrangement.

The illumination pattern generated by the temporally variable spatial light modulation arrangement comprises a plurality of illuminated or shadowed areas in the cross section of the illumination light beam path and on the sensor platform, which are also referred to as "illumination light pixels" below.

In addition, it is advantageous if a single element of the temporally variable spatial light modulation arrangement has a switching time of less than 20 msec for switching between positions or settings for forwarding to the sensor platform or blocking the illumination light.

In particular for measurements on cells immobilized on the sensor platform, it is advantageous if the arrangement for temporally rapidly variable spatial light modulation enables the generation of rapidly variable illumination patterns on the sensor platform, whereby temporally in their geometry changing objects than in their geometry time-varying measuring ranges the sensor platform can be specifically illuminated and light emanating from these objects can be detected.

The illumination system of the analytical system according to the invention may be one or more polychromatic or substantially comprise monochromatic light sources. It should be understood by a "substantially monochromatic" light source such a light source whose spectral emission width is less than 5 nm.

The one or more polychromatic or substantially monochromatic light sources may be selected from the group of lasers, vertical cavity surface emitting lasers VCSEL, edge emitting laser diodes, superluminescent diodes SLD, LED light emitting diodes, organic light emitting diodes (OLED), gas discharge lamps and incandescent lamps.

It is preferred that the illumination system in the optical beam path before entry of the illumination light into the arrangement for temporally variable spatial light modulation comprises optical or electro-optical components for producing a uniform over the illumination cross-section intensity distribution of the illumination light, these components are preferably selected from the group of optical projection systems , Microlens arrays, "light tunnels" and "light bars" or large-area emitting light sources with a variety of single light sources such as LEDs or OLEDs of different or the same emission wavelength, the total emission is formed using diffractive or refractive components to a homogeneous light distribution. Arrangements of large area homogeneous illumination systems with liquid crystal displays can also be used. In this case, the light of a rod-shaped light source is coupled into a planar light guide, in which the light is guided within the light guide by total internal reflection. On a surface of the light guide diffractive structures are applied, which selectively direct a portion of the light from the light guide. With a suitable choice of the diffractive structures acts Surface of the light guide as a nearly homogeneous light source. By means of a surface-mounted liquid crystal display, the light intensity of the homogeneous illumination can be spatially modulated. Such an arrangement is described, for example, in US Pat. No. 6,976,779.

"Light bars" or "light tunnels", such as LightTunnel ™ (Unaxis, Balzers, Liechtenstein), are used in commercially available projection systems and are based on the following functional principle: The light from one or more light sources is placed in one end of the "light tunnel" or "light bar" designated and designed as a hollow body, preferably cuboid or cylindrical optical component focused whose optical insides are mirrored. Here are referred to as "light bars" preferably cuboid or cylindrical body, which comprise a transparent material - usually glass or plastic - and in which light conduction by total internal reflection (TIR) occurs. After multiple reflections on the insides of such a "light tunnel" or "light bar", the light exiting at the opposite end side has an increased homogeneity of the intensity distribution over the illumination cross section.

In addition, it may be advantageous if telecentrically acting optical components are arranged in the optical beam path of the illumination light before the illumination light enters the arrangement for temporally variable spatial light modulation or in the further beam path in the direction of the sensor platform. Telescopic lenses, as described, for example, in US Pat. No. 6,324,016 (see FIGS. 1-6), enable the elimination of perspective perturbations and improved imaging of objects located outside the optical axis of an optical system. The disclosure of FIGS. 1-6 of US Pat. No. 6,324,016, together with the associated parts of US Pat Description hereby incorporated in full in the present application.

In the case of non-perpendicular, but a non-zero angle of incidence (measured from the surface normal of the substantially planar sensor platform) of the illumination light on the sensor platform, i. oblique light incidence, it may be advantageous if the optical system includes provisions for corrections to compensate for oblique incidence of light after Scheimpflug, as described for example in standard textbooks for photography.

The detection system of the analytical system according to the invention may comprise one or more detection units from the group of photodiodes, photomultipliers, avalanche diodes, CMOS arrays and CCD cameras.

It is preferred that the detection system comprises one or more spectrally splitting electro-optical components for a spectrally selective detection of the light emanating from the measurement areas. These may, for example, be spectrometers, spectrally selective optical filters such as short or long-pass filters, broadband optical filters or so-called notch filters adapted to laser emission lines.

It is preferred that the illumination system and / or the detection system comprise polarization-selective components in the beam path. It is particularly advantageous if the polarization-selective components allow the distinction between transversely electric (TE) and transverse magnetic (TM) polarized light, which emanates from the measuring ranges on the sensor platform. Furthermore, preferred embodiments of an analytical system according to the invention are those which are characterized in that the refractive index and / or the thickness of an adsorbed layer or its changes in or on the measurement areas on the sensor platform can be determined from the signals of the one or more detectors and their changes are. This may be, for example, the macroscopic refractive index of a liquid brought into contact with one or more measurement areas or its changes.

It is preferred that the light detected by the detection system be analyzed, wherein preferably the intensity or the intensity pattern is evaluated. For example, in the case of a resonance curve, the measured signal can be evaluated by a variety of methods known to those skilled in the art, as described, for example, in US Pat. No. 4,815,843 (column 6, last section, etc.). However, methods whose evaluation method and thus determined results are almost or even completely unaffected by the absolutely measured intensities on the detector are particularly preferred. In US 6429022 (Figure 12), an example is given of how any measured intensity pattern can be used. From this, a preferred embodiment of a method according to the invention for generating and measuring optical signals and / or their changes is derived, which is characterized in that with the aid of the detection system with at least one

Detection unit, a brightness distribution of the signals is measured, and that this brightness distribution is analyzed independently of measured absolute signal intensities (see also in a subsequent later in this description section). In general, when evaluating the light intensity measured by means of the detection system, methods are advantageous in which the sought measured variable, for example the position of a resonance curve, can be determined by methods in which the method preferred in the analysis hardly or not at all Intensity, but by a nearly or completely intensity-independent method can be determined, such as in the methods described below for determining the position of the resonance curve.

In the example of a resonance curve, a preferred method for determining the change in the position of the resonance is based on measuring the intensity on the flanks of the resonance curve. Using the first derivative of the resonance curve, the change in the position of the resonance curve can be calculated from the change in the intensity of the detected light. From the change in the position of the resonance curve, it is then possible, for example, to determine and calculate the change in the effective refractive index, as described, for example, in the prior art for the use of grating coupler sensor platforms.

Another preferred method for determining the change in the position of a resonance curve is based on the determination of the center of gravity of the measured resonance curve, which allows unambiguous conclusions about the position of the resonance curve. With a small change in the resonance position, the center of gravity changes accordingly.

Another preferred method for determining the position of a resonance curve is based on the use of a model curve, such as a Lorentz or Gaussian curve. Using numeric Method is adapted the model curve to the measured curve, which is also known as 'numeric fitting'. Commercial software for fitting model curves is offered, for example, by OriginLab (Northampton, MA 01060, USA). From the position of the model curve, the sought position of the measured resonance curve can be determined.

Such embodiments of an analytical system according to the invention, for example evanescent field sensor platforms, which make it possible to determine from the signals of the one or more detectors and / or their changes the effective refractive index and / or its changes in or on the measurement areas on the sensor platform are.

In this case, a possible embodiment is characterized in that from the signals of the one or more detectors and their changes, the resonance angle for coupling illumination light into a thin-film waveguide via a grid structured therein and / or changes such a resonance angle with respect to the surface normal of the sensor platform, with constant einstrahlter Wavelength of the illumination light, can be determined.

Another possible embodiment is characterized in that from the signals of the one or more detectors and their changes the resonant wavelength for coupling illumination light into a thin film waveguide over a grid structured therein and / or changes such a resonant wavelength, with constant irradiation angle of the illumination light with respect to the surface normal the sensor platform, are determinable. The irradiation of the illumination light and detection of the light emanating from the measurement areas can take place from opposite sides, relative to the surface of the sensor platform with the measurement areas located thereon. Preferably, however, the irradiation of the illumination light and the detection of the light emanating from the measurement areas take place on the same side of said surface.

The angle of incidence of the illumination light on the sensor platform can be in the range of -90 ° to + 90 ° (+ 90 ° and -90 ° where "parallel to the surface of the sensor platform"), with the angle range between -60 ° and + 60 ° is preferred. The illumination light may be polarized, for example transversely electrically (TE) or transversely magnetically (TM) polarized. In the case of TM polarized illumination light, an angle of incidence near the Brewster angle for the light entrance of the illumination light into the sensor platform on its surface side facing away from the measurement areas is particularly preferred since no Fresnel reflections from this interface of the sensor platform occur at this angle of incidence.

In the case of irradiation of the illumination light and detection of the light emitted by the measuring areas light on the same side with respect to the surface of the sensor platform with the measurement areas located thereon detection is rectified to the reflection direction of the illumination light, ie also in an angular range between -90 ° and + 90 °, an angle range between -60 ° and + 60 ° is preferred, and in the case of TM-polarized illumination light, detection below the Brewster angle is again particularly preferred.

Such a configuration of an analytical system according to the invention is illustrated in FIG. The illumination system 1 of the invention Analytical system consists of a lighting unit 8, which comprises a light source and an arrangement referred to as "SLM" for temporally fast variable spatial light modulation, and an illumination optical system 9. The illumination light 4 is according to this representation at an angle of incidence 6 of about 45 ° to the surface normal of The measuring areas (not shown) are located on the upper, opposite side surface of the sensor platform 2 with the curved arrows 12 and 12 'is supplied to the measuring areas and after interaction The detection angle 7 for detecting the light emanating from the sensor platform 2 or the measuring areas arranged thereon is equal to the reflection angle, that is about 45 ° in this case, chosen. The detection system 3 comprises a light collection optics 10, for example one or more collection lenses, and a detection unit 11.

An analogous to that in Figure 1 configuration, but in which

Detection system is not arranged in reflection, but in the transmission direction is shown in Figure 4.

FIG. 2 shows a preferred embodiment of an analytical system according to the invention. The illumination unit 8 comprises a light source 8.1, an electro-optical unit 8.2 for generating a homogeneous over the illumination cross-section intensity distribution of the illumination light. The electro-optical unit 8.2 preferably also comprises a telecentric optical imaging system. The homogenized in its intensity over the illumination cross section light is in the illumination beam path subsequently via a deflection mirror 8.3, in order to allow a compact design, directed in the designated as SLM arrangement 8.4 for temporally variable spatial light modulation. The use of "Digital Mirror Devices" DMD is advantageous for many applications because the polarization properties of the light are hardly affected by the mirror surface In the configuration of Figure 2, for example, a DMD from Texas Instruments Inc. (Piano, Texas, USA) is used The approximately 13 μm x 13 μm micromirrors of this DMD can be individually switched between + 12 ° and -12 ° in their position, with a speed of up to 16300 frames / sec with a DMD of 1024 x 768 pixels (individual mirrors) can be achieved.

In the optical path after the DMD (towards the sensor platform), an optical module consisting of prisms (referred to as "TIR prism") may be used to pass the light toward the illumination optics 9 (ON state of the illuminating light pixel) or TIR, which passes through the DMD as SML and an optional additional TIR-phsma, is further directed onto the sensor platform 2 by the illumination optics 9. Preferably the illumination optics 9 provide telecenter optical components which allow good control of the angle of incidence on the sensor platform 2 and the numerical aperture of the illuminating beam 10. Telecentric illumination optics may include lens systems or mirrors as described in US Patent 6,324,016.

The sensor platform can be illuminated at an angle of incidence of 0 °, ie parallel to the surface normal of the substantially planar sensor platform. In the case of a non-zero angle of incidence additional provisions for corrections may be made Compensation of the oblique incidence of light after Scheimpflug be provided in the illumination beam path.

In particular, for improving the signal-to-noise and / or the signal-to-background ratio of the measurement signals to be generated with the analytical system, filters such as polarization filters, spatial filters and spectral filters in the illumination beam path and / or in the detection beam path can additionally be used. Simultaneous use of two different polarizations (TE and TM polarization) allows additional information to be obtained, as first described in US Pat. No. 4,815,843 and later also in US Pat. No. 5,442,169 and in the application US 2005/0070027. For example, with the signals of thin film waveguides having gratings patterned therein, from the simultaneously or sequentially measured signals at TE and TM polarization, changes in the optical refractive index of the medium above the sensor platform and layer thickness changes at the surface of the sensor platform can be distinguished. To measure the two polarizations, the measured optical spectrum can be analyzed by calculating the expected spectrum for different polarizations with the aid of mathematical models and comparing them with the measured spectrum, which assigns the characteristic properties of the measured spectrum, such as " Peaks ", allowing for different polarizations. The polarizations can also be detected separately using polarizing components, such as

Polarizers or polarizing beam splitters, in the lighting system and / or detection system. Figure 6 illustrates the possibility of separating s- and p-polarized light by a polarizing beam splitter and then coupling the polarized light into optical fibers for subsequent separate detection of the spectra of the TE and TM polarized light. In a preferred Embodiment of an analytical system according to the invention, a measuring arrangement according to FIG. 8 from the US Pat. No. 4,815,843 is used, which is hereby introduced together with the associated part of the description as part of the present application.

For detecting the light emanating from the sensor platform, a combination of a light collecting optics 10 and a detection unit 11 is preferably used, as in the most general form in FIG. 1 for the detection in the direction rectified for reflection and in FIG. 4 for the detection in the transmission direction in FIG represented in the most general form.

At small angles of incidence 6 and detection angles 7, a beam splitter 9.1 is preferably used to separate the incident illumination light from the light emanating from the sensor platform and to illuminate the illumination light below the appropriate one

To direct irradiation angle on the sensor platform and to direct the outgoing of the sensor platform light to the detection unit, as shown in Figures 2, 5 and 6.

In a preferred embodiment of the analytical system according to the invention, as shown in FIG. 2, light emanating from the sensor platform 2 in the reflection direction is focused by a light collecting optics 10, for example an optical lens, onto one end of an optical fiber 10.1 and through this end face of the fiber coupled into this and then passed to the detection unit 11. Preferably, the detection unit 11 comprises a spectrometer, in which the collected light emanating from the sensor platform 2 can be spectrally analyzed.

The angular distribution of the light to be detected by the light-collecting optics 10 and emanating from the sensor platform 2 can be determined by a suitable choice of the optical properties, for example the numerical aperture Light collecting optics 10, modified and fixed. For sensor platforms based on refractive measurement principles, ie in particular for so-called label-free operating sensor platforms, narrow angular ranges, ie small aperture angles of the light cones, which are formed by the light emanating from the sensor platform 2, are preferred. In particular, light bundles with a narrow aperture angle can be optimally coupled into an optical fiber by the light beam is focused on an end face of the fiber, which at a distance from the focal length of the light-collecting optics 10, ie z. B. a converging lens is arranged.

The spectral properties of the light emanating from the sensor platform can be used to obtain information about the surface condition, in particular the surface coverage, of the sensor platform and thus also information about chemical, biochemical or biological processes, in particular binding or adsorption or desorption processes on the surface of the sensor Sensor platform, to get. The evaluation techniques and mathematical models required for this purpose are disclosed in the aforementioned patents and patent applications and known to the person skilled in the art also from his general knowledge and relevant textbooks. Known mathematical models are, for example, the "Rigorous Coupled Wave Analysis" (RCWA), which is often referred to as "Fourier Modal Method" (FMM), and MG Moharam and TK Gaylord, "Diffraction analysis of dielectric surface-relief gratings", J. Opt. Soc. Am., 72 (1997) 1385-1392, or in J. Turnen, "Diffraction theory of microrelief gratings", in Microoptics, HP Herzig, editor, Taylor's Francis Inc., (1997). Further mathematical models are the S and R matrices, which are described in detail in Lifeng Li, "Formulation and comparison of two recursive matrix algorithms for modeling layered diffraction gratings", J. Opt. Soc. At the. A, Vol. 13, 5, 1996, as well as the C-method described in Lifeng Li, Jean Chandezon, Gerard Granet, Jean-Pierre Plumey, "Rigorous and efficient grating-analysis method made easy for optical engineers", Appl. Opt., 38 (1999) 304-313, the method referred to as "Equivalent Source Method" which is described in AV Tischenko, M. Hamdoun, and O. Parriaux, "Two-dimensional coupled mode equation for grating waveguide excitation by a focused beam ", Opt. Quantum Electon. Special Issue on Workshop WTNM, Nottingham, 2001, as well as methods described in RH Morf, "Exponentially convergent and numerically efficient solution to Maxwell's equations of lammelar gratings", J. Opt. Soc. At the. A, Vol. 12, 1043-1056, 1995. The above models, methods and methods and their descriptions in the aforementioned publications are hereby incorporated in their entirety as part of the present application.

Another preferred embodiment of the analytical system according to the invention is shown in FIG. The optical components of the illumination system and of the detection system are largely identical to those of the embodiment according to FIG. The lighting unit comprises a light source 8.1 and a rear mirror 8 in the beam path, preferably formed as a parabolic mirror. The light emitted by the light source 8.1 light is homogenized by using a lens system 8.6 the light is focused on the face of a "light tunnel" or "light rod" 8.7. As a result of multiple reflections within the "light tunnel" or "light bar" 8.7, the intensity distribution of the light at its outlet opening is almost homogeneous and is detected by a prism system 8.8, preferably a TIR prism, on a spatial light modulator 8.4 as SMM, preferably a digital mirror device ( DMD), steered. A spatial filter 8.9 can be used for influencing the numerical aperture and the angular distribution of the light beams advancing in the direction of the sensor platform 2 before or after the spatial light modulator 8.4. A spatial filter consists essentially of an aperture diaphragm, which is introduced in the beam path. The light coming from the spatial filter 8.9 is essentially unpolarized, which is shown in FIG. 5 with two mutually perpendicular, solid arrows corresponding to the Lichtpolarisationsrichtungen within a circle, while the Hauptausbreitungsrichtung of the light is represented by an open arrow, which encloses the circle , The light is collimated by means of a lens system 9.2 and directed to a beam splitter 9.1, preferably a polarizing beam splitter, which preferably transmits only one polarization direction. The sensor platform 2, preferably an evanescent field sensor platform with grating structures, which have a periodicity of the grating lines formed in one or two dimensions, is illuminated with the preferably polarized light after the beam splitter 9.1. The grid lines of the sensor platform are aligned at an angle of preferably 45 ° to the polarization direction, which is indicated by the single filled double arrow, which symbolizes the polarization of the illumination light in front of the sensor platform in the illumination beam path. The light emanating from the sensor plate is polarized transversely (TE) and / or transversely magnetically (TM) after interaction with the grating waveguide structure and has a direction of polarization rotated by 45 ° relative to the illuminating light (indicated by in the drawing plane horizontal double arrow) and is referred to hereinafter as light emanating from the sensor platform or "signal light." The direction of polarization and the propagation direction of the light are each represented by the arrows in FIG.

Light which has undergone no interaction, for example by reflection or coupling into the grating waveguide structure, conduction in the waveguide and subsequent decoupling, is hereinafter referred to as "scattered light" designated. The scattered light generally has the same polarization as the irradiated illumination light.

The light components that have interacted with the sensor platform, i. in the case of a sensor platform based on a grating waveguide structure, in this application are referred to as the "outgoing from the sensor platform light".

The light emanating from the sensor platform is directed by the polarizing beam splitter 9.1 onto a lens system 10.2 as part of the light collection optics. In this case, only the "signal light" changed by its interaction with the sensor platform in its polarization direction is directed onto the lens system 10.2, while the essentially unchanged "scattered light" in its polarization direction is transmitted through the polarizing beam splitter 9.1. As a result, the "signal light" is separated from the "scattered light", which can lead to an improvement of the signal-to-noise ratio. This configuration is also often referred to as "crossed polarizers." The "signal light" is directed by means of a lens system 10.2 into an optical fiber 10.1 and guided to a light analysis unit 11.1, for example a spectrometer, as part of a detection unit. In the figure, a typical spectral curve (measured intensity l (λ) as a function of the wavelength λ), as may arise from reflection on a grating-waveguide structure, is shown schematically.

Another preferred embodiment of an analytical system according to the invention is shown in FIG. The optical components of the illumination system and of the detection system are largely identical to those of the embodiments according to FIGS. 2 and 5. The unpolarized light coming from the lens system 9.2 is directed onto the sensor platform 2 by means of a beam splitter 9.1. That of the Sensor platform 2 outgoing or reflected light is transmitted through the beam splitter 9.1 and by means of a lens system 10.2 and a deflecting mirror 10.3, through the use of which a more compact design can be realized, directed to a polarizing beam splitter 10.4. The polarizing beam splitter 10.4 separates the two

Polarization directions of the incident light on him. The two polarization directions are coupled separately into optical fibers 10.1, which guide the light to the light analysis unit 11.1, for example to several spectrometers for the detection of the spectra for different polarizations. In the figure, typical spectral curves, as may arise from reflection on a grating-waveguide structure, are shown schematically for TE and TM polarization.

Another preferred embodiment of an analytical system according to the invention is shown in FIG. The optical components of the illumination system and the detection system are largely identical to those of the embodiment of FIG. The embodiment according to FIG. 7 is characterized in that a prism for generating total internal reflection or a "resonant mirror" is used here as sensor platform 2.

FIG. 8 shows a further preferred embodiment of an analytical system according to the invention, in which the sensor platform 2 is designed as a microtiter plate or as its component. In addition, in this embodiment, the irradiation of the illumination light and the detection of the outgoing of the sensor platform light on opposite sides of the sensor platform, ie in a transmission light or transmitted light arrangement made.

Another embodiment of an analytical system according to the invention is characterized in that the illumination system has two or more arrangements for time-varying spatial light modulation. Such an embodiment is advantageously characterized by an increase in the spatial resolution of the illumination pattern to be generated on the sensor platform and / or by a further possible increase in the readout speed of the light signals emanating from the sensor platform, when the detection steps with the light modulation by the two or more SLMs are correlated. out. This embodiment is particularly preferred in combination with sensor platforms designed as microtiter plates.

Figures 9a and 9b show, using the example of a microtiter plate with 96 wells, possible variations of the illumination pattern generated on the sensor platform, which can be optimally adapted to the geometry of the measuring areas on the sensor platform. FIG. 9 a shows the illumination of a single measuring range, which here is equivalent to a single well of the microtiter plate. The illumination pattern is illustrated here by a hatched square around a measurement area (well) of the sensor platform denoted by "S." Figure 9b illustrates the simultaneous illumination (hatched cross) of 4 surrounding measurement areas (wells) which are used, for example, to obtain reference measurements and denoted by "R".

FIGS. 10a-10c illustrate how the illumination pattern to be generated on the sensor platform can be adapted to a wide variety of geometries. FIG. 10 a shows the illumination of a multiplicity of measurement areas in a hexagonal pattern. FIG. 10b shows the illumination of measuring areas which are arranged in concentric circular rings. FIG. 10c illustrates an illumination pattern with any irregular geometry, such as, for example, the targeted illumination of biological cells located on the sensor platform can correspond. In each case, the generated illumination pattern can be changed within a very short time by discrete switching of the individual elements of the SLM and, for example, adapted to changes in the geometry of the measuring ranges.

- An optical system with an illumination system for illumination of measurement areas on the sensor platform, with a designated as "SLM" arrangement for temporally fast variable spatial light modulation and with a detection system with at least one detection unit for detecting signals from the measurement areas on the sensor platform in the direction the transmission or reflection of the illumination light in a spectral range that includes the spectral range of the illumination light ,. and

a sensor platform which can be inserted into the optical system and has measuring areas arranged thereon in a one- or two-dimensional array, characterized in that, in the operating state, illumination patterns emerge from an illumination light entering this SLM with a substantially homogeneous intensity distribution in the cross section of the illumination light perpendicular to its propagation direction Selectable and quickly changeable geometry that can be determined by the settings of the SLM can be generated on the sensor platform. In this case, a method is preferred which is characterized in that a brightness distribution of the signals is measured with the aid of the detection system with at least one detection unit, and that this brightness distribution is analyzed independently of measured absolute signal intensities.

Particular preference is given to such an embodiment of the method according to the invention, which is characterized in that the optical signals and / or their changes are generated from discrete measurement ranges by one or more binding or adsorption events between one or more analytes in one or more samples and specific Recognition elements for said analytes in or on said measurement areas, wherein the samples and the detection elements for the analyte to be detected in the samples are brought into contact with each other on the measurement areas, and that from these optical signals and / or their changes a simultaneous qualitative and / or quantitative Detection of a variety of analytes in one or more samples and / or one or more analytes in a variety of samples is possible.

Another object of the present invention is the use of an analytical system according to one of the aforementioned embodiments and / or a method according to one of the aforementioned embodiments for quantitative and / or qualitative analyzes for the determination of chemical, biochemical or biological analytes in screening methods in the pharmaceutical research, the combinatorial Chemistry, clinical and preclinical development, real-time binding studies and kinetic parameters in affinity screening and research, qualitative and quantitative analyte determinations, in particular for DNA and RNA analysis and the determination of genomic or proteomic differences Genome, such as single nucleotide polymorphisms, for the measurement of protein-DNA interactions, for the determination of control mechanisms for mRNA expression and for protein synthesis, for the preparation of toxicity studies and for the determination of expression profiles, in particular for the determination of biological and chemical marker substances, such as mRNA, proteins, peptides or low-molecular organic (messenger) substances, as well as for the detection of antibodies, antigens, pathogens or bacteria in pharmaceutical product research and development, human and veterinary diagnostics, agrochemical Product research and development, the symptomatic and presymptomatic plant diagnostics, for patient stratification in pharmaceutical product development and for the therapeutic drug selection, for the detection of pathogens, pollutants and pathogens, in particular Salmonella, prions, viruses and bacteria, especially in d he food and environmental analysis.

In the following examples, the present invention will be further explained without limiting the generality.

Examples

example 1

The structure of the analytical system corresponds to the embodiment of Figure 4, i. Arrangement of the lighting system and the

Detection system on opposite sides of the sensor platform and detection of the transmitted light component. According to a preferred embodiment of such an arrangement, the illumination system 1 comprises a commercial projection system with light emitting diodes (LEDs) as light sources. Suitable for this purpose For example, a projection system of the type FF1 (Toshiba, Taiwan). FIG. 12 shows the emission spectrum of this projector with simultaneous operation of all three LED light sources. The blue, green and red LEDs used have central emission wavelengths of 460 nm, 520 nm and 627 nm, respectively, and a spectral emission bandwidth of typically 20 nm to 40 nm. In the following examples, the red light source is mainly used, with, of course, also light sources of different emission spectrum , for example with blue or green emission, can be used. The light of the LEDs is homogenized by means of a "light tunnel" and directed to a Digital Mirror Device (DMD) as SLM The light reflected by the DMD is directed by a first lens system contained in the projection system With the aid of an additional lens (focal length f = 500 mm As part of the illumination optics 9, the light coming from the projector is imaged onto the sensor platform, which is located at a distance of about 1 m from the projector For the purpose of checking or determining the polarization of the illumination light, the illumination unit in the illumination beam path comprises a polarizer.

The optical sensor platform comprises a thin film waveguide having a grating structure structured therein, i. a structure known as the "resonant lattice structure" comprising a planar glass substrate as support having a refractive index of 1.5 at 633 nm and having two large parallel surfaces at a distance of 1 mm as the thickness of the support in which one of the two mentioned

Surface is structured an optical grating with a period of 360 nm. Preferably, this grid is pronounced as a surface relief grid with mutually parallel webs and grooves of the same web width or groove width, wherein the distance between two successive webs or grooves is referred to as the grating period. The grid can be over the entire surface over an entire surface of the glass substrate be educated. The grating is covered with a substantially optically transparent layer having a refractive index of 2.1 at 633 nm and a thickness of 150 nm as waveguiding layer.

As is known to those skilled in the art, for coupling light into the high-refractive-wave waveguiding layer of such a grating-waveguide layer, there is a discrete resonance angle for light coupling in the forward direction of light propagation in the waveguide and a resonance angle for light coupling in the backward direction of light propagation in the waveguide upon irradiation of illumination light of a discrete wavelength and a certain polarization. For example, the resonance angles for light coupling of TM and TE polarized light with 626 nm wavelength can differ by the order of 10 °. For illumination light of different wavelengths, the resonance angles are again different. Upon irradiation of illumination angles at a fixed angle of incidence, but with a certain spectral width of, for example, 1 nm to 10 nm, therefore, the wavelength of the light component for which the resonance condition for the light coupling is met at the predetermined irradiation angle is referred to as the "resonance wavelength ".

With a suitable choice of the spectral width of the light source and the physical characteristics (in particular layer thickness, grating period and lattice depth) of the "resonant lattice structure", it is possible to use the

To meet resonance conditions for light coupling both in the backward direction and in the forward direction of the light propagation in the waveguide simultaneously. The center of the two resonances corresponds to the autocollimation angle, which corresponds to the perpendicular to the surface plane of the sensor platform. Thus, can be next to Determining the measured variable also determine the orientation of the sensor platform in the region of the measuring range.

Furthermore, with a suitable choice of the spectral width of the light source and the characteristics of the "resonant grating structure", the resonance conditions for light coupling for both directions of polarization, ie for TE and TM polarization, can be satisfied simultaneously, which can be simultaneously described later in more detail , the thickness and the refractive index of a layer applied to the surface of the sensor platform, for example an adhesion-promoting layer, a complete or partial, ie perforated, layer of detection elements for analyte detection or of bound analyte compounds.

The simultaneous fulfillment of the resonance conditions for light coupling for both directions of propagation and for both polarization directions can of course also be realized in combination.

From changes in the resonance angle and the resonance wavelength, changes in the effective refractive index can be determined, which are caused, for example, by molecular adsorption or desorption processes on the surface of the sensor platform and corresponding changes in the (macroscopic) refractive index at this surface. Because of the well-known

Dependence of the refractive index on the layer and light parameters can be determined from these changes in the effective refractive index or of the resonance angle or wavelength as coupling parameters of the sensor platform, for example absolute or relative changes in the mass coverage on the surface of the sensor platform. If a calibration of these changes with known If mass assignments (for example with complete or partial monolayers of surface adsorbed molecules of known molecular weight) are or are present, it is also possible to determine absolute values of the surface coverage. In particular, with known density of the surface coverage can be drawn in the case of non-spherical surface adsorbed molecules from the height of the signal changes, for example, conclusions about the orientation of the adsorbed molecules. This even makes it possible to draw conclusions about the nature of the corresponding chemical, biochemical or biological adsorption or binding process. Conversely, for simulated adsorption processes, corresponding changes in the coupling parameters can be simulated and the models for these simulated adsorption processes can then be checked using real measured values.

Simultaneous measurement for two different polarizations allow mathematical models to determine the refractive index and thickness of a surface deposited layer. This additional information allows further conclusions. For example, it can be determined whether molecules have attached to the surface as a dense monolayer or have accumulated loosely and in multiple layers. In this case it is also easier to draw conclusions about the orientation of non-spherically symmetric molecules, since different orientations of the molecules on the surface, for example for "lying" molecules and for "stationary" molecules, with the same mass assignment to different optical properties of the sensor layer, in particular to different refractive indices and layer thicknesses lead.

According to the embodiment according to the configuration of FIG. 4, the detection unit is aligned in the transmission direction along the optical axis of the illumination light generated by the illumination unit, in this case, the detection angle θ coincides with the incident angle θ of the illumination light on the sensor platform. As the light collecting optics 10, a lens with a focal length of 500 mm is used in this example, with which the light is focused on the entry end face of an optical fiber, from where the light component thus coupled into the fiber and guided therein is guided into a spectrometer for further spectral analysis. In this case, the lens is arranged at a distance of approximately 500 mm in each case between the sensor platform and the optical fiber.

FIG. 12 shows, for four differently set angles of incidence 6.3, 6.4, 6.5 and 6.6 of the illumination light on the sensor platform, the spectra recorded in transmission of the TE-poled illumination light of the red LED. At the resonance wavelength for the light coupling corresponding to the respective angle of incidence, the spectrum of the transmitted light has a minimum. From the position, width and depth, it is possible, with known physical parameters of the sensor platform, to draw conclusions about the effective refractive index or adsorption or desorption processes on the sensor platform. Corresponding spectra for TM-polahsiertes illumination light at two different angles of incidence 6.1 and 6.2 are shown in Figure 13, in which case the transmission minima are less pronounced at the respective resonance wavelengths.

Example 2

The construction of the analytical system corresponds to the embodiment according to FIG. 3, ie arrangement of the illumination system and the detection system on the same side of the sensor platform and detection in the direction of the reflected light component. According to a preferred Embodiment of such an arrangement, the lighting system 1 comprises a commercial projection system with light-emitting diodes (LEDs) as light sources. Suitable for this purpose is, for example, a projection system of the type FF1 (Toshiba, Taiwan). FIG. 11 shows the emission spectrum of this projection system with simultaneous operation of all three LED light sources. The blue, green and red LEDs used have central emission wavelengths of 460 nm, 520 nm and 627 nm and a spectral emission bandwidth of typically 20 nm to 40 nm. The light from the LEDs is homogenized by means of a "light tunnel" and onto a digital mirror device (DMD) as SLM 4. The light reflected by the DMD is directed by a first lens system included in the projection system.

With the help of an additional lens (focal length f = 500 mm) as part of the illumination optics 9, the light coming from the projector is on the

Sensor platform, which is located at a distance of about 1 m from the projector. In order to check or fix the polarization of the illumination light, the illumination unit in the illumination beam path comprises a polarizer.

The optical sensor platform comprises a thin-film waveguide with a lattice structure structured therein, ie a structure known as the "resonant lattice structure" comprising a planar glass substrate as support with a refractive index of 1.5 at 633 nm and with two large parallel surfaces at a distance of 1 mm as the thickness of the carrier, in which one of the two surfaces mentioned is structured an optical grating having a period of 360 nm Groove width is pronounced, where the grating period is the distance between two successive webs or grooves can be formed over the entire surface over an entire surface of the glass substrate. The grating is covered with a substantially optically transparent layer having a refractive index of 2.1 at 633 nm and a thickness of 150 nm as waveguiding layer.

According to the embodiment according to the configuration of FIG. 3, the detection unit is aligned in the reflection direction along the optical axis of the illumination light generated by the illumination unit, in which case the detection angle 7 coincides in magnitude with the incident angle 6 of the illumination light on the sensor platform, but with opposite sign , As the light collecting optics 10, a lens with a focal length of 500 mm is used in this example, with which the light is focused on the entry end face of an optical fiber, from where the light component thus coupled into the fiber and guided therein is guided into a spectrometer for further spectral analysis. In this case, the lens is arranged at a distance of approximately 500 mm in each case between the sensor platform and the optical fiber.

FIG. 14 shows, for a single set angle of incidence of the illumination light on the sensor platform, the spectrum of the TM-poled illumination light of the red LED recorded in the reflection direction. At the resonance wavelength corresponding to this angle of incidence for the light coupling, the spectrum has a sharp maximum in the reflection direction. From the position, width and depth, it is possible, with known physical parameters of the sensor platform, to draw conclusions about the effective refractive index or adsorption or desorption processes on the sensor platform. List of reference numbers:

The following list of reference numerals is part of the disclosure of this patent application.

1: Lighting system

2: sensor platform

3: detection system

4: illumination light 5: light 6 originating from the sensor platform; 6.1; 6.2; 6.3;

6.4; 6.5; 6.6: Incidence angle on the sensor platform

7: detection angle

8: Lighting unit

8.1: light source

8.2: Electro-optical unit for generating an over the

Illumination cross section homogeneous distribution of the

illumination light

8.3: Deflecting mirror

8.4: Arrangement for temporally quickly changeable

Light modulation (SLM)

8.5: Mirror

8.6: lens system

8.7: "Light Tunnel" or "Light Bar"

8.8: prism system

8.9: Spatial filter

9: Illumination optics

9.1: beam splitter

9.2: lens system

10: Light collection optics

10.1: Optical fiber 10.2: lens system

10 .3: Deflection mirror

1 1 detection unit

1 1 .1: Light analysis unit i 1 1 .2: Polarizing beam splitter

12 Sample, test, calibration or reference supplies in liquid or gaseous form supplied to the sensor platform

12 ': sample, test, calibration or reference supplies in liquid or gaseous form removed from the sensor platform

Claims

54Patentansprüche

1. Analytical system for generating and measuring optical signals and / or their changes from measuring ranges, which are arranged in a one- or two-dimensional array on a substantially optically transparent sensor platform, at least comprising - an optical system with an illumination system for illuminating measuring ranges on the sensor platform and a detection system with at least one detection unit for

Detection of signals from the measurement areas on the sensor platform, in the direction of the transmission or reflection of the illumination light in a spectral range which includes the spectral range of the illumination light, and - a sensor platform insertable into the optical system with measurement areas arranged in a one- or two-dimensional array, characterized in that the illumination system comprises an arrangement referred to as "SLM" for temporally rapidly variable spatial light modulation, with the illumination mode in the operating state from an entering into this SLM illumination light, with substantially homogeneous intensity distribution in the cross section of the illumination light perpendicular to its propagation direction, illumination pattern and rapidly changeable geometry that can be determined by the settings of the SLM can be generated on the sensor platform.

2. Analytical system according to claim 1, characterized in that the sensor platform in a one- or two-dimensional arrangement comprises more than 50, preferably more than 500, most preferably more than 50,000 measuring ranges. 55

3. Analytical system according to one of claims 1 - 2, characterized in that the sensor platform in a single or multi-layer system comprises a material from the group consisting of substantially optically transparent glasses, plastics and

Ceramics is formed, wherein layers of these materials may optionally be provided with additional coatings.

4. Analytical system according to one of the preceding claims, characterized in that the sensor platform as a

Evanescent field sensor platform is formed.

5. Analytical system according to claim 4, characterized in that the sensor platform is selected from the group of prisms for generating total internal reflection, self-supporting optical

Waveguides, optical thin-film waveguides, thin-film waveguides with grids structured therein for the light coupling and / or light outcoupling, resonant lattice structures and "resonant mirrors".

6. Analytical system according to one of claims 1-5, characterized in that the sensor platform has the basic dimensions of a microtiter plate or is formed as part of a microtiter plate.

7. Analytical system according to one of claims 1-6, characterized in that the arrangement for temporally variable spatial light modulation is selected from the group of "Digital Mirror Devices" DMD, liquid crystal displays LCD, "Liquid Cristal on Silicon Silicon" LCOS microdisplays and mechanical 56

movable masks with translucent and light-blocking areas.

8. Analytical system according to claim 7, characterized in that the arrangement for temporally variable spatial

Light modulation a variety discretely for forwarding to the sensor platform or blockade of the illumination light switchable individual elements in a one- or two-dimensional arrangement, wherein a measuring range of more than one of these individual elements is illuminated.

9. Analytical system according to claim 8, characterized in that the arrangement for temporally variable spatial light modulation a plurality discretely switchable for forwarding to the sensor platform or blockade of the illumination light

Comprises individual elements in a two-dimensional arrangement, which is preferably more than 100 x 100 discretely switchable individual elements.

10. Analytical system according to one of claims 8 or 9, characterized in that a single element of the arrangement for temporally variable spatial light modulation has a switching time of less than 20 msec for the change between positions or settings for forwarding the illumination light to the sensor platform or blockade of the illumination light ,

11. Analytical system according to one of claims 1-10, characterized in that the arrangement for temporally rapidly variable spatial light modulation allows the generation of rapidly variable illumination patterns on the sensor platform, which temporally in their geometry is changing 57

Objects as in their geometry time-varying measuring ranges on the sensor platform specifically illuminated and emitted by these objects outgoing light can be detected.

12. Analytical system according to one of claims 1-11, characterized in that the illumination system comprises one or more polychromatic or substantially monochromatic light sources.

13. An analytical system according to claim 12, characterized in that the one or more polychromatic or substantially monochromatic light sources are selected from the group of lasers, vertical cavity surface emitting lasers VCSEL, edge emitting laser diodes, superluminescent diodes SLD, light emitting diodes LED, organic light-emitting

Diodes (OLED), gas discharge lamps and incandescent lamps.

14. Analytical system according to one of claims 1-13, characterized in that the illumination system in the optical beam path before entering the illumination light in the arrangement for temporally variable spatial light modulation optical or electro-optical components for producing a homogeneous over the illumination cross-section intensity distribution of the illumination light, wherein these components are preferably selected from the group of optical projection systems,

Microlens arrays, "light tunnels" and "light rods".

15. Analytical system according to one of claims 1-14, characterized in that the illumination system in the optical beam path of the illumination light before the occurrence of the illumination light in the arrangement for temporally variable spatial light modulation 58

or comprises in the further beam path in the direction of the sensor platform telecentrically acting optical components.

16. Analytical system according to one of claims 1-15, characterized in that the optical system comprises provisions for corrections for compensation of oblique incidence of light after Scheimpflug.

17. Analytical system according to one of claims 1-16, characterized in that the detection system comprises one or more detection units from the group of photodiodes, photomultipliers, avalanche diodes, CMOS arrays and CCD cameras.

18. Analytical system according to one of claims 1-17, characterized in that the detection system comprises one or more spectrally splitting electro-optical components for a selective detection of spectral properties of the emanating from the measurement areas light.

19. Analytical system according to one of claims 1-18, characterized in that the illumination system and / or the

Detection system polarization-selective components in the beam path include.

20. Analytical system according to claim 19, characterized in that the polarization-selective components enable the distinction between transversely electric (TE) and transversely magnetic (TM) polarized light emanating from the measuring areas on the sensor platform.

21. Analytical system according to one of claims 1-20, characterized in that from the signals of the one or more 59

Detectors and their changes, the refractive index and / or the thickness of an adsorbed layer or their changes in or on the measuring ranges on the sensor platform can be determined.

22. Analytical system according to one of claims 4 - 21, characterized in that from the signals of the one or more detectors and / or their changes, the effective refractive index and / or its changes in or on the measuring ranges on the sensor platform can be determined.

23. Analytical system according to claim 22, characterized in that from the signals of the one or more detectors and their changes the resonance angle for coupling of illumination light in a thin-film waveguide via a grid structured therein and / or changes such a resonance angle with respect to

Surface normals of the sensor platform, at constant irradiated wavelength of the illumination light, can be determined.

24. Analytical system according to claim 22, characterized in that from the signals of the one or more detectors and their

Changes the resonance wavelength for coupling of illumination light in a thin film waveguide on a grid structured therein and / or changes of such a resonant wavelength, at a constant angle of incidence of the illumination light with respect to the surface normal of the sensor platform can be determined.

25. A method for generating and measuring optical signals and / or their changes from measurement areas, which in a one- or two-dimensional array on a substantially optically transparent 60

Sensor system platform are arranged, using an analytical system according to one of the preceding claims, comprising at least - an optical system with an illumination system for illumination of measurement areas on the sensor platform, with an arrangement known as "SLM" for temporally fast variable spatial light modulation and with a detection system with at least one detection unit for detecting signals from the measurement areas on the sensor platform, in the direction of the transmission or reflection of the illumination light in a spectral range which includes the spectral range of the illumination light, and

a sensor platform which can be inserted into the optical system and has measuring areas arranged thereon in a one- or two-dimensional array, characterized in that, in the operating state, an illumination light entering this SLM with a substantially homogeneous intensity distribution in the cross section of the illumination light perpendicular to its propagation direction illuminates freely selectable and quickly changeable geometry that can be defined by the settings of the SLM

Sensor platform can be generated.

26. The method according to claim 25, characterized in that with the aid of the detection system with at least one detection unit, a brightness distribution of the signals is measured, and that this brightness distribution is analyzed independently of measured absolute signal intensities.

27. The method according to any one of claims 25 - 26, characterized in that the optical signals and / or their changes from discrete 61

Measuring ranges are generated by one or more binding or Adsorptionsereignisse between one or more analytes in one or more samples and specific recognition elements for said analytes in or on said measuring ranges, wherein the samples and the detection elements for the analyte to be detected in the samples on the measuring areas with each other from these optical signals and / or their changes, a simultaneous qualitative and / or quantitative detection of a plurality of analytes in one or more samples and / or one or more analytes in a plurality of samples is made possible.

28. Use of an analytical system according to one of claims 1 - 24 and / or a method according to one of claims 25 - 27 for quantitative and / or qualitative analyzes for the determination of chemical, biochemical or biological analytes in screening methods in the pharmaceutical research, the combinatorial chemistry, clinical and preclinical development, real-time binding studies and kinetic parameters in affinity screening and research, qualitative and quantitative analyte determinations, in particular for DNA and RNA analysis and the determination of genomic or proteomic differences in the genome, such as single nucleotide Polymorphisms, for the measurement of protein-DNA interactions, for the determination of control mechanisms for m-RNA expression and for protein synthesis, for the preparation of toxicity studies and for the determination of expression profiles, in particular for the determination of biolog and chemical marker substances, such as mRNA, proteins, peptides or low-molecular organic (messenger) substances, and for the detection of antibodies, antigens, pathogens or bacteria in pharmaceutical product research and development, human and veterinary diagnostics, agrochemical product research and development, the 62

symptomatic and presymptomatic plant diagnostics, for patient stratification in pharmaceutical product development and for therapeutic drug selection, for the detection of pathogens, pollutants and pathogens, in particular Salmonella, prions, viruses and bacteria, in particular in food and environmental analysis.