Multianalyte determination system and methods

• US 20040038386A1
• Filed: 03/04/2003
• Published: 02/26/2004
• Est. Priority Date: 09/04/2000
• Status: Active Grant

• Current Assignee: Bayer Technology Services GmbH (Bayer AG)
• Original Assignee: Bayer Technology Services GmbH (Bayer AG)
• Inventors: Pawlak, Michael, Bopp, Martin Andreas, Zesch, Wolfgang
• Granted Patent: US 7,879,598 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B

Abstract

The invention relates to embodiments of an optical system for luminescence determination, comprising two or more excitation light sources, a sensor platform and an optical component with several discrete facets for beam deflection towards the sensor platform. Further subjects of the invention are methods for luminescence determination with an optical system according to the invention and analytical systems, as well as the use of these methods for quantitative affinity sensing and for various other applications. The aim of the present invention is to provide optical and analytical measuring devices for highly sensitive detection of one or more analytes, using a multitude of measurement areas on a common carrier.

Claims

1. An optical system for luminescence determination, comprising at least two excitation light sources, a sensor platform and an optical component with several discrete facets for deflecting a light beam, wherein the angle of divergence between excitation light falling onto different facets of said optical component is increased or reduced by at least a factor of 1.2 in the optical path departing from said optical component, in comparison to the original divergence angle (between said light rays irradiated onto said different facets).

2. An optical system according to claim 1, wherein the angle of divergence between excitation light falling onto different facets of said optical component is increased by at least a factor of 1.2, preferably by at least a factor of 1.5, in the optical path departing from said optical component, in comparison to the original divergence angle (between said light rays irradiated onto said different facets).

3. An optical system according to any of claims 1-2, wherein said optical component with several discrete facets for beam deflection is a multi-facet mirror with planar or curved facets, preferably with planar facets.

4. An optical system according to any of claims 1-2, wherein said optical component with several discrete facets for beam deflection is a multi-facet prism with planar or curved facets, preferably with planar facets.

5. An optical system according to any of claims 1-4, wherein the light from two or more excitation light sources of similar or different wavelength falls onto the same facet of said optical component.

6. An optical system according to any of claims 1-4, wherein a dedicated facet of said optical component is provided for each different excitation wavelength, the excitation light of said excitation wavelength to be directed onto the corresponding dedicated facet.

7. An optical system according to any of claims 1-6, wherein the beam deflection of excitation light of different wavelength into different predefined directions occurs with an offset of less than 0.2 mm between the centers of the deflected beams on the sensor platform.

8. An optical system according to any of claims 4-7, wherein the multi-facet prism comprises additional means for deflecting or masking reflections of the excitation light emanating from the sensor platform.

9. An optical system according to any of claims 4-8, wherein one or more reflective facets of the multi-facet prism are partly or completely metallized.

10. An optical system according to any of claims 1-9, wherein additional optical elements for the spectral selection of the excitation wavelength, such as interference filters or edge filters, and optionally additional optical elements for beam attenuation, such as optical neutral density or grey filters, optionally provided as a “

continuously varying”

filter with a continuous local gradient of the transmission, and/or further elements for beam guiding, such as glass fibers, optionally connected to micro lenses or diffractive optical elements, are located in the optical path of the excitation light between the at least one excitation light source and the optical component with several discrete facets for beam deflection.

11. An optical system according to any of claims 1-10, wherein two or more lasers with different emission wavelengths are used as excitation light sources.

12. An optical system according to any of claims 1-11, wherein additional optical elements comprising, for example, diffractive optical elements and/or lenses for beam expansion and/or for generation of a parallel beam and/or diaphragms or masks for partial masking of the beam are located in the optical path of the excitation light between the light sources and the sensor platform, in order to generate a desired beam profile on the sensor platform.

13. An optical system according to any of claims 1-12, wherein the sensor platform comprises a multitude of discrete measurement areas, in which biological or biochemical or synthetic recognition elements for the determination of one or more analytes are immobilized.

14. An optical system according to any of claims 1-13, wherein up to 100,000 measurement areas are provided in a two-dimensional arrangement on the sensor platform.

15. An optical system according to any of claims 1-14, wherein a single measurement area has an area of 0.001 mm²-6 mm².

16. An optical system according to any of claims 1-15, wherein an adhesion-promoting layer is deposited between the biological or biochemical or synthetic recognition elements and the sensor platform.

17. An optical system according to claim 16, wherein the adhesion-promoting layer has a thickness of less than 200 nm, preferably of less than 20 nm, and wherein the adhesion-promoting layer preferably comprises a chemical compound of the groups comprising silanes, epoxides, functionalized, charged or polar polymers, and “

self-organized passive or functionalized mono- or double-layers”

.

18. An optical system according to any of claims 1-17, wherein laterally separated measurement areas (d) are generated on the sensor platform by laterally selective deposition of biological or biochemical or synthetic recognition elements on said sensor platform.

19. An optical system according to claim 18, wherein, as said biological or biochemical or synthetic recognition elements, components of the group formed by nucleic acids (e.g. DNA, RNA, oligonucleotides) and nucleic acid analogues (e.g. PNA), mono- or polyclonal antibodies, peptides, enzymes, aptamers, synthetic peptide structures, soluble membrane-bound proteins and proteins isolated from a membrane, such as receptors, their ligands, antigens for antibodies, “

histidin-tag components” and

their complex forming partners, cavities generated by chemical synthesis, for hosting molecular imprints, etc.

20. An optical system according to claim 18, wherein whole cells, cell components, cell membranes or their fragments are deposited as biological or biochemical or synthetic recognition elements.

21. An optical system according to any of claims 18-20, wherein regions between the laterally separated measurement areas are “

passivated”

for minimization of non-specific binding of analytes or their tracer compounds, i.e., that compounds, that are “

chemically neutral”

towards the analyte, are deposited between the laterally separated measurement areas (d), preferably for example out of the groups formed by albumins, especially bovine serum albumin or human serum albumin, casein, unspecific polyclonal or monoclonal, alien or empirically unspecific antibodies for the one or the multiple analytes to be determined (especially for immuno assays), detergents—

such as Tween 20®

—

fragmented natural or synthetic DNA not hybridizing with polynucleotides to be analyzed, such as extract from herring or salmon sperm (especially for polynucleotide hybridization assays), or also uncharged but hydrophilic polymers, such as poly ethyleneglycols or dexiranes.

22. An optical system according to any of claims 1-21, wherein the luminescence light from the measurement areas on the sensor platform is directed onto at least one opto-electronic detector.

23. An optical system according to any of claims 1-22, wherein the luminescence light from the measurement areas is imaged onto a locally resolving detector, which is preferably selected from the group formed by CCD cameras, CCD chips, photodiode arrays, avalanche diode arrays, multi-channel plates and multi-channel photomultipliers.

24. An optical system according to any of claims 1-23, wherein the luminescence light from the measurement areas is imaged onto the at least one opto-electronic detector by means of a system comprising one or more lenses and/or mirrors.

25. An optical system according to any of claims 1-24, wherein one or more optical elements for selection of the emission wavelength and discrimination of light of other wavelengths, such as diffractive elements, interference filters or edge filters, are provided in the emission beam path between the sensor platform and the at least one opto-electronic detector for recording the luminescence light emanating from the measurement areas.

26. An optical system according to claim 25, wherein the emission beam path has a divergence or convergence of less than 15°

at the position of application of said optical element for the spectral selection.

27. An optical system according to any of claims 25-26, wherein the optical elements for selection of the emission wavelength and for discrimination of light of other wavelengths, such as interference filters or edge filters, are located between the two halves of a tandem objective.

28. An optical system according to any of claims 1-27, wherein the sensor platform comprises an optically transparent support (supporting substrate), preferably of glass or a thermoplastic plastics, on which the biological or biochemical or synthetic recognition elements are immobilized in the measurement areas.

29. An optical system according to any of claims 1-28, wherein the sensor platform comprises a planar optical waveguide.

30. An optical system according to claim 29, wherein the sensor platform comprises an optical thin-film waveguide with a first optically transparent layer (a) on a second optically transparent layer (b), and wherein the optically transparent layer (b) has a lower refractive index than the optically transparent layer (a).

31. An optical system according to any of claims 29-30, wherein the in-coupling of excitation light into the optically transparent layer (a) is performed using a diffractive grating (c) modulated in the layer (a), which is preferably a relief grating.

32. An optical system according to claim 31, wherein grating structures (c) provided in the waveguiding layer (a) of the sensor platform are arranged in a one- or two-dimensional array, with even, non-modulated regions of the waveguiding layer (a) being adjacent to the grating structures in the direction of propagation of an excitation light to be in-coupled into the layer (a), and wherein arrays of two or more measurement areas are provided on these non-modulated regions, which can optionally additionally be fluidically sealed against each other in discrete sample compartments.

33. An optical system according to any of claims 31-32, wherein second grating structures (c′

) for the out-coupling of excitation light and, where appropriate, of luminescence light back-coupled into the waveguiding layer (a) are provided on the sensor platform, in addition to grating structures (c) for the in-coupling of excitation light, in order to out-couple again the light guided in the waveguiding layer (a), after its passing through the region of the measurement areas (in direction of propagation of the guided excitation light following an in-coupling grating structure (c)).

34. An optical system according to any of claims 31-33, wherein grating structures (c) and optionally (c′

) are provided discretely for individual segments (arrangements in one- or two-dimensional arrays) of measurement areas.

35. An optical system according to any of claims 31-33, wherein grating structures (c) and optionally (c′

) are provided as continuous strips (columns) extending over the whole sensor platform perpendicular to the direction of propagation of the excitation light to be in-coupled.

36. An optical system according to any of claims 33-35, wherein grating structures (c′

) are also used as in-coupling gratings (c) upon sequential performance of measurements.

37. An optical system according to any of claims 33-36, wherein grating structures (c) for the in-coupling and (c′

) for the out-coupling of light out of the waveguiding layer (a) of the sensor platform have the same period and are modulated continuously below all measurement areas of the sensor platform.

38. An optical system according to any of claims 30-37, wherein the product of the thickness of layer (a) and its refractive index is one tenth up to a whole, preferably one third to two thirds of the excitation wavelength of an excitation light to be coupled into layer (a).

39. An optical system according to any of claims 30-38, wherein a thin metal layer, preferably of gold or silver, optionally on an additional dielectric layer with lower refractive index than layer (a), for example of silica or magnesium fluoride, is deposited between the optically transparent layer (a) and the immobilized biological or biochemical or synthetic recognition elements, wherein the thickness of the metal layer and of the optional additional intermediate layer is selected in such a way that surface plasmon can be excited at the excitation and/or the luminescence wavelength.

40. An optical system according to any of claims 1-39, wherein optically or mechanically recognizable marks for simplifying adjustments in an optical system and/or for the connection to sample compartments as part of an analytical system and/or as helps for a later image analysis are provided on the sensor platform.

41. An optical system according to any of claims 1-40, wherein the sensor platform is provided with a mark, such as a barcode.

42. An optical system according to any of claims 1-41, wherein the excitation light from the two or more light sources is essentially monochromatic.

43. An optical system according to any of claims 1-42, wherein the excitation light from different light sources is irradiated simultaneously from different directions towards the sensor platform in such a way, that the offset between the beam centers on the sensor platform is less than 0.2 mm.

44. An optical system according to any of claims 1-43, wherein optical components of the group comprising lenses or lens systems for the shaping of the transmitted light bundles, planar or curved mirrors for the deflection and optionally additional shaping of the light bundles, prisms for the deflection and optionally spectral separation of the light bundles, dichroic mirrors for the spectrally selective deflection of parts of the light bundles, neutral density filters for the regulation of the transmitted light intensity, optical filters or monochromators for the spectrally selective transmission of parts of the light bundles, or polarization selective elements for the selection of discrete polarization directions of the excitation or luminescence light are located between the sensor platform and the one or more detectors.

45. An optical system according to any of claims 1-44, wherein the excitation light is launched in pulses with a duration of 1 fsec to 10 min.

46. An optical system according to any of claims 1-45, wherein the emission light from the measurement areas is measured time-resolved.

47. An optical system according to any of claims 1-45, wherein for referencing purposes light signals of the group comprising excitation light at the location of the light sources or after expansion of the excitation light or after its multiplexing into individual beams, scattered light at the excitation wavelength from the location of the one or more laterally separated measurement areas, and light of the excitation wavelength out-coupled by the grating structure (c) besides the measurement areas are measured.

48. An optical system according to claim 47, wherein the measurement areas for determination of the emission light and of the reference signal do partly or completely overlap and are preferably identical.

49. An optical system according to any of claims 33-48, wherein the optimization of the adjustment for optimum in-coupling of excitation light by means of an in-coupling grating (c) towards measurement areas provided in direction of propagation of the in-coupled light is performed upon maximization of the excitation light out-coupled by an out-coupling grating (c′

) and measured by a detector, wherein this optimization is preferably performed under computer control.

50. An optical system according to claim 49, wherein a rotation of the optical component with several discrete facets for beam deflection, around an axis located inside or outside of said optical component, is performed for the optimization of the adjustment of the coupling angle.

51. An optical system according to claim 50, wherein the optical component with several discrete facets for beam deflection is connected with a rotary element with axis of rotation inside or outside of said optical component in such a way, that the offset of the beam on the sensor platform is less than 0.3 mm upon a rotation of said optical component around said axis of rotation by less than 5°

.

52. An optical system according to any of claims 49-51, wherein a translation of the sensor platform in parallel or perpendicular to the grating lines is performed for optimization of the coupling position.

53. An optical system according to any of claims 49-51, wherein the optimization of the adjustment is performed upon maximization of one or more reference signals from one or more measurement areas on the sensor platform, wherein this optimization is preferably performed under computer control.

54. An optical system according to claim 53, wherein said reference signal is scattered light of the excitation wavelength.

55. An optical system according to claim 53, wherein said reference signal is luminescence light from measurement areas dedicated for purposes of referencing and/or of adjustment.

56. An optical system according to any of claims 13-55, wherein the irradiation of the excitation light to and detection of emission light from one or more measurement areas is performed sequentially for one or more measurement areas.

57. An optical system according to claim 56, wherein the sensor platform is moved between steps of sequential excitation and detection.

58. An analytical system, for the determination of one or more analytes in at least one sample on one or more measurement areas on a sensor platform by luminescence detection, with

an optical system according to any of claims 1-57

supply means for bringing the one or more samples into contact with the measurement areas on the sensor platform.

59. An analytical system according to claim 58, wherein said analytical system additionally comprises one or more sample compartments, which are at least in the area of the one or more measurement areas or of the measurement areas combined to segments open towards the sensor platform.

60. An analytical system according to claim 59, wherein the sample compartments have a volume of 0.1 nl-100 μ

l each.

61. An analytical system according to any of claims 59-60, wherein the sample compartments are closed, except for inlet and/or outlet openings for the supply or outlet of samples, at their side opposite to the optically transparent layer (a), and wherein the supply or the outlet of the samples and optionally of additional reagents is performed in a closed flow-through system, wherein, in case of liquid supply to several measurement areas or segments with common inlet and outlet openings, these openings are preferably addressed row by row or column by column.

62. An analytical system according to any of claims 59-60, wherein the supply of the samples and optionally of additional reagents is performed in parallel or crossed micro-channels, upon exposure to pressure differences or to electric or electromagnetic potentials.

63. An analytical system according to any of claims 59-61, wherein the sample compartments are provided with openings at the side facing away from the optically transparent layer (a), for locally addressed supply or removal of the samples or of other reagents.

64. An analytical system according to any of claims 59-63, wherein the sample compartments are arranged in an array, comprising the sensor platform as the base plate and a body combined therewith in such a way, that an array of cavities is generated between the base plate and said body, for generation of an array of flow cells fluidically sealed against each other, and that at least one outlet of each flow cell leads to a reservoir fluidically connected with said flow cell and capable to receive liquid exiting from said flow cell.

65. An analytical system according to claim 64, wherein the reservoir for receiving liquid exiting from the flow cell is provided as a recess in the exterior wall of the body combined with the base plate.

66. An analytical system according to any of claims 58-65, wherein said analytical system comprises 2-2000, preferably 2-400, most preferably 2-100 sample compartments.

67. An analytical system according to any of claims 58-66, wherein the pitch (geometrical arrangement in rows and/or columns) of the inlets of the sample compartments does correspond to the pitch (geometrical arrangement) of the wells of a standard microtiter plate.

68. An analytical system according to any of claims 58-66, wherein the arrangement of sample compartments with the sensor platform as the base plate and the body combined therewith does correspond to the footprint of a standard microtiter plate.

69. An analytical system according to any of claims 58-66, with, for example, 2 to 8 sample compartments in a column or, for example, 2 to 12 sample compartments in a row, wherein said sample compartments in a column or row themselves are combined with a carrier (“

meta-carrier”

) with the dimensions of standard microtiter plates in such a way, that the pitch (geometrical arrangement in rows and/or columns) of the inlets of the flow cells does correspond to the pitch (geometrical arrangement) of the wells of a standard microtiter plate.

70. An analytical system according to claims 58-69, wherein a removable bottom protection is provided below the sensor platform as the base plate of an arrangement of sample compartments, and wherein optionally the upper side of the arrangement of sample compartments is closed with an additional covering top, for example a film, a membrane or a cover plate.

71. An analytical system according to claim 70, wherein the bottom protection is removed automatically or semi-automatically before a measurement is started.

72. An analytical system according to any of claims 58-71, comprising an optical system according to any of claims 30-57, wherein at least one grating structure (c) modulated in the waveguiding layer (a) of a sensor platform as the base plate, for the in-coupling of excitation light towards the measurement areas, is provided within each sample compartment.

73. An analytical system according to any of claims 58-71, comprising an optical system according to any of claims 33-57, wherein grating structures (c) are provided within the range of the sample compartments and additional grating structures (c′

) for light out-coupling are always arranged outside of those sample compartments where the in-coupling is performed

74. An analytical system according to any of claims 58-71, with an optical system according to any of claims 33-57, wherein grating structures (c) and optional additional grating structures (c′

) extend over the range of several or of all sample compartments.

75. An analytical system according to any of claims 58-74, wherein 5-5000, preferably 10-400 measurement areas are provided in one sample compartment.

76. An analytical system according to any of claims 58-75, wherein additionally mechanical means and a transport mechanism are provided, operable for an automated transport of an arrangement of sample compartments, comprising a sensor platform as a base plate and a body combined therewith, from a location of the insertion of that arrangement to the location of luminescence excitation and detection and optionally from there back to the original position.

77. An analytical system according to any of claims 58-76, wherein said analytical system additionally comprises a receiving device (“

stacker”

) for receiving a plurality of arrangements of sample compartments.

78. An analytical system according to claim 77, wherein the loading of the “

stacker”

from the position of the insertion of the arrangement of sample compartments and the transport of said arrangement of sample compartments from there to the location of luminescence excitation and detection and then back to the original location is performed automatically.

79. An analytical system according to any of claims 58-78, wherein said analytical system comprises one or more temperature-controllable zones.

80. An analytical system according to claim 79, wherein the arrangement of sample compartments and/or the excitation light sources and/or the one or more opto-electronic detectors and/or the “

stacker”

can be temperature-controlled separately.

81. An analytical system according to any of claims 77-80, wherein the arrangement of sample compartments and/or the excitation light sources and/or the one or more opto-electronic detectors and/or the “

stacker”

are operated under a higher air pressure than ambient pressure.

82. An analytical system according to any of claims 58-81, wherein said analytical system additionally comprises one or more electronic control components for control of the status of one or more optical or electrical or mechanical components, which control components can generate an optical or acoustic or electronic alarm signal if necessary.

83. An analytical system according to any of claims 58-82, wherein said analytical system comprises means for preventing the insertion of not adequate sensor platforms, for example with wrong mechanical dimensions.

84. An analytical system according to any of claims 58-83, wherein said analytical system additionally comprises one or more electronic processors, connected to storage media and electronic connecting media, a keyboard for data or command input, a screen and a program code for automated operation.

85. An analytical system according to any of claims 58-84, wherein the operation of said analytical system and/or the measurement are performed automatically using pre-defined files for initialization.

86. An analytical system according to any of claims 58-85, wherein said analytical system is reset automatically to a pre-defined initial status and possibly generated measurement data are secured in a file, when an indicated error function has occurred.

87. An analytical system according to any of claims 58-86, wherein a file is generated automatically for each measurement, in which file are stored the code of the used sensor platform, the essential measurement parameters and the measurement data.

88. An analytical system according to any of claims 58-87, wherein local variations of the excitation light intensity on the sensor platform and/or of the detection sensitivity of the optical system for light signals from different positions on the sensor platform are corrected using means which comprise, for example, the recording of images for correction taken at the excitation wavelength and/or at one or more luminescence wavelengths, the calculation of theoretical distributions of the available excitation light intensity, theoretical calculations of the locally resolved efficiency of the optical imaging and detection system, etc.

89. A method for the determination of one or more analytes by luminescence detection, upon using an analytical system according to any of claims 55-88 which comprises an optical system according to any of claims 1-57, wherein one or more liquid samples to be analyzed for the one or more analytes are brought into contact with one or more measurement areas on the sensor platform, excitation light is directed towards the measurement areas, thus exciting compounds capable of luminescence in the sample or on the measurement areas to luminescence and the emitted luminescence is measured.

90. A method according to claim 89, wherein for generation of luminescence at least one luminescent dye or luminescent nanoparticle is used as a luminescence label, which can be excited and emits at a wavelength between 300 nm and 1100 nm.

91. A method according to any of claims 89-90 characterized in that it comprises means to extend the dynamic range for signal recording by at least a factor of 3.

92. A method according to claim 91, wherein the means for extending the dynamic range comprise the application of differently long exposure times, i.e., the duration of the irradiation of the excitation light and the integration time of the detector, which exposure times can be varied by at least a factor of 3.

93. A method according to claim 91, wherein the means for extending the dynamic range comprise a variation of the excitation light available on the sensor platform by at least a factor of 3, for example upon using discrete neutral density filters in the excitation beam path, optionally provided as a “

continuously varying”

filter with a continuous local gradient of the transmission, or upon variation of the intensity of the light sources or upon changing the adjustment of the sensor platform with respect to the excitation beam path.

94. A method according to any of claims 89-93, wherein the at least one luminescence label is bound to the analyte or, in a competitive assay, to an analyte analogue or, in a multi-step assay, to one of the binding partners of the immobilized biological or biochemical or synthetic recognition elements or to the biological or biochemical or synthetic recognition elements.

95. A method according to any of claims 89-93, wherein a second or more luminescence labels of similar or different excitation wavelength as the first luminescence label and similar or different emission wavelength are used.

96. A method according to claim 95, wherein charge or optical energy transfer from a first luminescent dye acting as a donor to a second luminescent dye acting as an acceptor is used for the detection of the analyte.

97. A method according to any of claims 89-96, wherein the one or more luminescences and/or determinations of light signals at the excitation wavelengths are performed polarization-selective.

98. A method according to any of claims 89-97, wherein the one or more luminescences are measured at a polarization that is different from the one of the excitation light.

99. A method according to any of claims 89-98 for the simultaneous or sequential, quantitative or qualitative determination of one or more analytes of the group comprising antibodies or antigens, receptors or ligands, chelators or “

histidin-tag components”

, oligonucleotides, DNA or RNA strands, DNA or RNA analogues, enzymes, enzyme cofactors or inhibitors, lectins and carbohydrates.

100. A method according to any of claims 89-99, wherein the samples to be examined are naturally occurring body fluids, such as blood, serum, plasma, lymph or urine, or egg yolk or optically turbid liquids or tissue fluids or surface water or soil or plant extracts or bio- or process broths, or are taken from biological tissue fractions or from cell cultures or cell extracts.

101. The use of an optical system according to any of claims 1-57 and/or of an analytical system according to any of claims 58-88 and/or of a method according to any of claims 89-100 for the quantitative or qualitative analyses for the determination of chemical, biochemical or biological analytes in screening methods in pharmaceutical research, combinatorial chemistry, clinical and pre-clinical development, for real-time binding studies and for the determination of kinetic parameters in affinity screening and in research, for qualitative and quantitative analyte determinations, especially for DNA- and RNA analytics, for generation of toxicity studies and for the determination of gene and protein expression profiles, and for the determination of antibodies, antigens, pathogens or bacteria in pharmaceutical product development and research, human and veterinary diagnostics, agrochemical product development and research, for symptomatic and pre-symptomatic plant diagnostics, for patient stratification in pharmaceutical product development and for the therapeutic drug selection, for the determination of pathogens, nocuous agents and germs, especially of salmonella, prions and bacteria, in food and environmental analytics.