DE102015001033A1 - High-throughput screening system for performing optical measurements - Google Patents

Abstract

The present invention relates to a high-throughput screening system for performing optical measurements, in particular spectroscopy. The object of the invention to provide a high-throughput screening system for carrying out spectroscopic measurements, in particular for Ramam spectroscopy, which enables all spectra of all microtiter plate wells to be simultaneously and rapidly measured in the wells is achieved by the high-throughput screening system comprises a laser and optical path excitation unit, an optical fiber fiber optic fiber bundle readout unit and a spectrometer, a microtiter plate having individual sample wells, and a microtiter plate well and positioning unit for positioning the microtiter plate wells Fiber bundle, wherein the fiber bundle is a dimensionally reducing fiber bundle, which on its side facing the sample a rectangular or square arrangement of the optical fibers and on its side facing the spectrometer a line-shaped arrangement having the optical fibers, and that an optical highly multimode illumination fiber is multiplied over a lens and a microlens array multiplied in an intermediate image plane and can then be imaged via two further lenses in the plane of the sample, all cavities simultaneously through the excitation and readout unit can be illuminated and metered with a radiation in which the radiation to be generated can be transmitted via a system of optical lenses from the cavities to the line-shaped fiber bundle and from this into the spectrometer.

Description

The present invention relates to a high-throughput screening system for performing optical measurements, in particular spectroscopy.

High-throughput screening

High-throughput measurements using microtiter plates are widely used in biology, medicine and pharmacy. The use of vending machines allows more than 100,000 samples a day, depending on the application.

High-throughput measurements are performed for different analysis methods.

In optical methods for the investigation of scattered radiation, the scattering cross section and the maximum power which can be used without destroying the sample decide on the measurement time required for a container (one well) of the microtiter plate. The sample containers (cavities) are filled by machines with several pipettes at the same time. The reading is often done in a grid, so that each container (cavity) is read out individually.

Also, Raman spectroscopic investigations are generally suitable for the investigation of various properties of substances. So revealed the WO 2013/064159 A1 For example, a method for the determination of nuclear DNA in sperm and the DE 102 37 394 A1 discloses, for example, a method for determining mechanical properties of polymer products. In this case, a Raman spectrum is recorded by at least one sperm or a polymer product and from the Raman spectrum of the at least one sperm / polymer product, the at least one biochemical / mechanical property of the sperm / polymer product is calculated, so that a fast, non-invasive, reproducible and providing a biochemical / mechanical characterization of sperm / polymer product without sample preparation.

In addition, from the EP 1 463 392 A1 such as EP 1 651 934 A1 Method for the detection of pathogenic microorganisms, from the document EP 1 766 350 A1 spectroscopic method for the analysis of component particles, from EP 1 766 351 A1 a multimodal procedure for the identification of dangerous substances and of the WO 2012/165837 A3 an arrangement and a method for high-throughput screening by means of Raman spectroscopy with core-cap-shell nanoparticles for drug discovery known. A disadvantage of the technical solution according to WO 2012/165837 A3 is that the use of filters only poor or no spectra can be detected.

In order to keep the measuring time as low as possible for high-throughput measurements using microtiter plates, it is necessary to read several containers (wells) of the microtiter plate at the same time. The spectra must be read out as quickly as possible and without any time delay. For this it is necessary to maximize the lighting performance and to transmit as much of the scattered radiation as possible. For the use of the greatest possible illumination performance, it is necessary to illuminate the sample containers (the wells of the microtiter plate) homogeneously and in their area as far as possible. For this, incoherent lighting is an advantage. In addition, the illumination for each sample container (each well of the microtiter plate) must be identical. The walls of the microtiter plate should not be illuminated, so the generation of unwanted scattered radiation is avoided. In addition, depending on the spectroscopic analysis method, it may be advantageous to irradiate the sample with depolarized light, thereby also allowing for easy polarization adjustment via a linear polarizing filter. In order to ensure the comparability of the spectra of the individual sample containers, a standardization must be carried out.

To use different sized microtiter plates, the system must be easily adjustable to this.

Due to the known screening methods, spectra can only be recorded individually, which (depending on the examined radiation) leads to very long measuring times and leads to the fact that with temporally variable samples no simultaneous examination of the individual containers (cavities) is possible. In addition, the individual containers (wells of the microtiter plate) are not homogeneously illuminated by a simple focusing in the sample plane, whereby the maximum sample performance is severely limited. Conventional wide-field methods lead to significantly worse results than those achieved when using a spectrometer. An imaging of this kind is also superfluous and annoying, since only individual containers (cavities) and no complete chemical images are to be read.

A disadvantage of the known technical solutions is that not all spectra can be measured simultaneously and quickly in the cavities and the excitation power for all cavities and the intensity of the transmitted scattered radiation (in particular in Raman spectroscopy ) are not high.

The invention is therefore based on the object to provide a high-throughput screening system for performing spectroscopic measurements, in particular for Raman spectroscopy, which avoids the disadvantages of the prior art, in particular allows all spectra of the wells of a microtiter plate simultaneously can be quickly measured in the cavities.

According to the invention this object is achieved by the characterizing features of the first and the sixth claim. Further favorable embodiments of the invention are specified in the subordinate claims.

The essence of the invention is that in the high-throughput screening system for performing optical measurements in spectroscopy, a dimension-reducing fiber bundle for receiving / detecting the spectroscopic radiation and a special illumination and imaging system are used.

For this purpose, the high-throughput screening system comprises a laser and optical path excitation unit, a fiber optic fiber optic bundle readout unit and a spectrometer, a microtiter plate with individual sample wells, and a microtiter plate microtiter plate well positioning and positioning unit opposite the fiber bundle.

It is essential that the fiber bundle is a dimension-reducing fiber bundle, which has on its side facing the sample a square array of optical fibers and on its side facing the spectrometer a line-shaped arrangement of the optical fibers, and that an optical highly multimode illumination fiber via a lens and a microlens array is multiplied in an intermediate image plane and can then be imaged via two further lenses in the plane of the sample, wherein all cavities can be simultaneously illuminated and measured by the excitation and readout unit with a radiation in which the producible radiation to be measured via a system of optical lenses from the cavities to the fiber bundle and from this into the spectrometer is transferable.

The illumination fiber has a large length in the range of several decameters and is provided with a vibration device, so that the polarization and coherence of the laser-emissive radiation in the fiber by the vibrator and the length of the fiber is destructible, such that the sample can be irradiated with uniformly unpolarized light or when using a linear polarizing filter with linearly polarized light of any polarization direction.

It is also within the scope of the invention that, in addition to the illumination fiber provided with a vibrator, illumination times at a CCD are used which are large in relation to the inverse vibration frequency, thus destroying the coherence of the illumination of the sample and the illuminated areas of the sample with equal intensity and homogeneity can be illuminated.

The optical path leads from the laser via the optical illumination fiber and a line filter, so that the scattered radiation generated in the fiber is removable.

The fiber bundle consists of light conducting fibers of angular or circular cross section, between which are nonconductive intermediate fibers of round cross section which space the photoconductive fibers on the side facing the spectrometer.

The photoconductive fibers on the sample-facing side are arranged at a distance from each other in the array, so that only the light generated in the samples and not the light emanating from the walls of the microtiter plate is received.

The high throughput screening system is designed to operate by irradiating the samples in the wells of the microtiter plates simultaneously with uniformly unpolarized light or by using an additional linear polarizer with linearly polarized light in which the laser produces monochromatic radiation is passed over the illumination fiber and the line filter, so that in the illumination fiber, the scattered radiation generated in the fiber is removed, wherein simultaneously with the vibration device on the illumination fiber, a vibration is generated, so that the coherence of the radiation emanating from the laser in the illumination fiber through the vibration device is destroyed.

The excitation light generated in this way is imaged on the samples in the cavities such that the illuminated surfaces, after imaging on the fiber bundle, coincide in their extent with the fibers of the fiber bundle, so that homogeneous and intensity-identical illumination surfaces are produced in the sample-containing wells of the microtiter plate.

The radiation emitted by the samples is read out with the fiber bundle.

Subsequently, a computer-aided standardization of the read-out data with a homogeneously scattering sample is carried out in order to obtain accurate quantitative statements with regard to the intensities of the scattered radiation emanating from the samples.

The light can also be widened before entering the illumination fiber and controlled by a perforated panel in the power.

Within the scope of the invention there is also the possibility that the microtiter plate is moved during the irradiation in such a way that a larger sample area is illuminated per cavity and thus a higher maximum possible illumination power is enabled, resulting in a shorter necessary exposure time or a higher ratio of signal and noise is possible.

Moreover, by replacing the lens between the illumination fiber and the microtiter plate, it is also possible to adjust the high-throughput screening system very quickly and automatically to different microtiter plates without the functionality of the illumination thereby changing.

The method can also be used for Raman spectroscopy, fluorescence spectroscopy, absorption spectroscopy, transmission spectroscopy or for the combination of these types of spectroscopy.

It is also possible for several lasers to be coupled into the illumination fiber in order to be able to carry out measurements at different excitation wavelengths merely by changing the filters, otherwise the illumination remains constant, or that several light sources are coupled into the illumination fiber at the same time, so that the Probe is excited by these light sources simultaneously, and the radiation emitted by the sample is examined, or that the fiber-based sample illumination is performed not by the incident light method, but by another lens, or in the transmitted light method.

The method can be used, for example, for the simultaneous recording of at least two spectra for the calculation of difference spectra based on the simultaneously measured reference spectrum and for the analysis of the smallest spectral shifts in optical spectroscopy.

The invention will be explained in more detail below with reference to the embodiments and the figures. Showing:

1 FIG. 2 shows a schematic representation of an embodiment of the high-throughput screening system according to the invention for carrying out optical measurements in spectroscopy, FIG.

2 FIG. 2: a representation of the typical intensity distribution recorded with a beam profile camera, as in a single cavity using the system according to FIG 1 occurs

3 : a schematic representation of a section of the illumination according to 1 in the sample plane with a square illumination fiber (The filled squares represent the illumination with monochromatic radiation. The dashed squares symbolize the fibers of the fiber bundle, as they are illuminated by the image of the scattered radiation),

4 FIG. 2: a schematic representation of the dimension-reducing fiber bundle according to FIG 1 and

5 FIG. 2: a representation of the polarization dependence of the transmission of an illumination fiber according to FIG 1 30 m long with coupled radiation with a wavelength of 532 nm (The triangles represent the polarization dependence of the laser.) The squares are related to the measurement with the illumination fiber For comparison, a comparison measurement with a shorter fiber was taken Measurements are drawn as circles.).

This in 1 The high-throughput screening system for carrying out optical measurements in spectroscopy comprises an excitation unit with a laser ( 1 ) and an optical path with an illumination fiber ( 6 ), a readout unit with a fiber bundle ( 23 ) of optical fibers and a spectrometer ( 25 ), a microtiter plate ( 18 ) with cavities for individual samples and a movement and positioning unit for the arrangement of the wells of the microtiter plate ( 18 ) relative to the fiber bundle ( 23 ).

The optical path runs from the laser ( 1 ) via a pinhole ( 3 ), a lens ( 4 ) or via a lens in the front side ( 5 ) of the illumination fiber ( 6 ), so that the light from the laser ( 1 ) into the illumination fiber ( 6 ) can be coupled.

Between the laser ( 1 ) and the pinhole ( 3 ) can also be a telescope ( 2 ) can be arranged.

The head of the illumination fiber ( 6 ) is located just behind the focal plane of the lens ( 4 ), so that a focus formation in the illumination fiber ( 6 ) is prevented.

The illumination fiber ( 6 ) is a visually highly multimode fiber that passes through the lens ( 4 ) and a microlens array is multiplied in an intermediate image plane and can then be imaged via two further lenses in the plane of the sample, all cavities simultaneously illuminated by the excitation unit with a radiation and read by the readout unit (reading the spectroscopic radiation), in which the producible radiation to be measured via a system of optical lenses from the cavities to the line-shaped fiber bundle ( 23 ) and from this into the spectrometer ( 25 ) is transferable.

Starting from the laser ( 1 ) as a monochromatic light source, the excitation light is transmitted through the lens ( 4 ) or via a lens in the front side ( 5 ) of the illumination fiber ( 6 ) coupled. The fiber head is just behind the focal plane of the lens ( 4 ) to prevent focus formation in the fiber.

In front of the lens is a pinhole ( 3 ). This serves for the fast, simple and temporally stable adjustment of the power with which the sample is irradiated.

To increase the adjustable power range can be before the aperture ( 3 ) another telescope ( 2 ). The different focus diameter at the focal point of the lens following this power adjustment ( 4 ) as well as the changing numerical aperture during the coupling into the fiber head ( 5 ) of the illumination fiber ( 6 ) only affect the power with which the sample is excited. All other properties of the lighting remain constant.

For the illumination fiber ( 6 ) is a highly multimode optical fiber with a length of several decimeters. The length is crucial for the generation of depolarized light. The two end faces ( 5 ) and ( 8th ) have the same shape. The front side ( 5 ) is the laser ( 1 ) facing side and the front side ( 8th ) is the laser ( 1 ) side facing away.

The illumination fiber ( 6 ) has a core with a dimension which ensures the guidance, one for the homogenization of sufficient number of modes.

The shape of the core (for example, circular or square) determines the shape of the illuminated area of a sample-fillable well of the microtiter plate ( 18 ). The cores of the fibers of the fiber bundle ( 23 ) have the same shape as the core of the illumination fiber ( 6 ). However, the dimensions can be chosen differently.

About the coupling of the illumination fiber ( 6 ) to a vibration device ( 7 ) this is put into a temporal oscillation. The oscillation leads to the temporal superimposition of the interference in the illumination fiber ( 6 ), so that the coherence for exposure times greater than a multiple of the period of the oscillation disappears. As a result, interference patterns are avoided and there is a homogeneous power distribution on the illuminated surfaces.

The illumination fiber ( 6 ) itself generated by the excitation with that of the laser ( 1 ) outgoing radiation an interference signal. This signal is transmitted through a line filter ( 14 ), which covers all wavelengths except those of the monochromatic laser ( 1 ) blocked, filtered out.

The monochromatic excitation light is starting from the end face ( 8th ) of the fiber via an image consisting of the lens ( 9 ) and the lens array ( 10 ), in an intermediate level ( 11 ). The front side ( 8th ) of the illumination fiber ( 6 ) is located at the focal point of the lens ( 9 ) and the intermediate level ( 11 ) arises at the focal point of the lens array ( 10 ).

The number of lenses of the microlens array ( 10 ) is equal to the number of fibers in the fiber bundle ( 23 ) to choose.

For optimum utilization, the lenses of the microlens array ( 10 ) a square shape.

In the intermediate level ( 11 ), corresponding to the number of lenses in the microlens array ( 10 ), a certain number of images of the fiber end surface ( 8th ), which all have the same intensity and homogeneous distribution.

The intermediate layer ( 11 ) is transmitted through the lens ( 12 ) and the lens ( 17 ), or the objective, formed infinity microscope on the plane of the microtiter plate ( 18 ).

The microtiter plate ( 18 ) has a very large number of similar cavities. The number of cavities should be at least as large as the number of fibers in the fiber bundle (FIG. 23 ) be.

About the lens ( 17 ) and the lens ( 21 ) is the radiation to be examined on the end face of the fiber array ( 22 ) / of the fiber bundle ( 23 ). The focal lengths of the lenses ( 9 ) 10 ) 12 ) 17 ) and ( 21 ) are chosen so that the illuminated sample surfaces after imaging on the end face of the fiber array ( 22 ) / fiber bundle in position and dimension with the cores of the fibers on the end face of the fiber array ( 22 ) to match. The focal lengths are chosen so that the maximum numerical aperture of the radiation to be examined can be transmitted.

The numerical aperture of the illumination is transmitted through the aperture ( 13 ) adjusted so that, depending on the size of the wells of the microtiter plate ( 18 ) it does not come to the stimulation of the material of the microtiter plate.

Within the beam path of the component ( 12 ) and ( 17 ), the monochromatic excitation light is transmitted through a beam splitter, such as a dichroic mirror ( 16 ) at an angle of 45 ° or by an edge filter at a very low angle, which is usually less than 10 °, reflected. The filter ( 16 ) transmits radiation having a wavelength greater than that of the excitation wavelength and reflects radiation having a wavelength less than or equal to the excitation wavelength. By using a rotatable linear polarizing filter ( 15 ), the sample surface ( 18 ) are irradiated with linearly polarized light.

By removing or folding out, this polarizing filter ( 15 ), the sample is stored in the microtiter plate ( 18 ) was irradiated with unpolarized light.

In the beam path of the scattered light, formed by the lens ( 17 ) and lens ( 21 ), for polarization-dependent measurements, a linear polarization filter ( 20 ).

For suppressing the Rayleigh radiation and the excitation light reflected from the sample, which is emitted by the laser ( 1 ), an additional edge filter ( 19 ). The fiber array ( 27 ) is located in the focal plane of the lens ( 21 ).

By changing the lens ( 17 ), the excitation beam path and the beam path of the scattered light are changed to the same whiteness, so that only those passing through the fiber array ( 22 ) imaged sample surfaces over the laser ( 1 ). The system can be easily adapted to different microtiter plates.

The fiber array ( 22 ) consists of a fiber bundle ( 23 ) of identical multimode fibers, which have a defined distance from each other. The fiber bundle ( 23 ) for reading out the samples is a dimension-reducing fiber bundle, which on its side facing the sample a rectangular or square arrangement of the optical fibers and on its side facing the spectrometer ( 24 ) [= Front side] has a line-shaped arrangement of the optical fibers. This front page ( 24 ) is located exactly in the cleavage plane of the spectrometer ( 25 ). In the fiber bundle ( 23 ), other fibers may be interlaced which do not conduct light. These intermediate fibers serve as spacers on the front side ( 24 ). Between each two fibers of the fiber array is then in each case at least one intermediate fiber on the front side ( 24 ).

This avoids crosstalk due to imaging in the spectrometer.

The technical specifications and the goal of maximizing the accumulated numerical aperture sometimes results in the fact that only a relatively small area of each sample can be illuminated. In order to be able to use a higher power for illumination, in order thereby to obtain smaller exposure times or a larger ratio of signal and noise, the microtiter plate ( 18 ) are moved so automatically during the recording, so that a total of a larger amount of each sample illuminated and thus damage to the sample is avoided. In this case, a homogeneous sample distribution within a cavity is assumed, and a (lateral) average value of the sample content is measured by means of the movement method.

In the following, the embodiment of the hand 1 to 5 even more concrete:
When using the light source in 1 is a frequency-doubled Nd: YAG laser that emits a TEM ₀₀ mode at 532 nm. The laser is transmitted through a biconvex lens ( 4 ) with a focal length of 2 cm into the illumination fiber ( 6 ) coupled. The cross-section of the core and cladding of the illumination fiber is square. The side length of the core is 160 microns and that of the shell 200 microns. The lighting fiber is 30 meters long and set in FC connectors. The length is sufficient to generate almost completely depolarized light.

To illustrate this fact, the intensity at the end of a 30 m long fiber was measured by means of an analyzer. In 5 the dependence on the analyzer position is shown. The measured values shown as filled circles refer to a reference measurement with a square fiber of only 2 m in length. The measured values shown as solid triangles show the polarization-dependent intensity of the laser without fiber optic illumination. All measured values are normalized for better comparability.

As vibration device ( 7 ) a commercial toothbrush is used. In 2 is shown an image of the end surface of a lighting fiber. In this example, a fiber with a core side length of 400 μm is chosen. The measurement was carried out with a beam profile camera. The Standard deviation of the intensity on the plateau is always less than 2%.

The fiber end surface ( 8th ) is located in the center of the focal plane of a lens ( 9 ) with a focal length of 8 cm.

The microlens array ( 10 ) consists of 64 square lenses with a focal length of 1 cm and a lens spacing of 1 mm. In the focal plane ( 11 ) of the lens array, 64 identical images of the fiber end face ( 8th ) with a total side length of 8 mm. The individual images each have a side length of 20 μm.

About the lens ( 12 ) with a focal length of 3 cm and the lens ( 17 ) with a focal length of 3 cm, the intermediate image ( 11 ) transferred to the sample level.

In 3 is a section of a microtiter plate ( 18 ) shown schematically (not to scale). The wells of the microtiter plate ( 222 ) have a distance of 1 mm to each other. The filled areas ( 333 ) set by the laser ( 1 ) illuminated areas. These squares have a side length of 20 microns. With this lighting, the walls ( 111 ) of the microtiter plate ( 18 ) not excited to a depth of 5 mm.

The scattered light is transmitted through the lens ( 17 ) and the lens ( 21 ), which has a focal length of 12 cm imaged on the fiber array (see 1 ).

The dashed squares ( 444 ) in 3 symbolize the fibers of the fiber bundle as they are illuminated by the image of scattered radiation. The beam path of the scattered light is separated from that of the excitation light by a dichroic mirror ( 16 ) separated. This reflects light with a wavelength less than or equal to 532 nm. Light with a longer wavelength is transmitted. After the dichroic filter is an edge filter ( 19 ), which transmits only light with a wavelength greater than 532 nm (see 1 )

In 4 are the fiber bundle ( 2222 ) and its end faces ( 1111 and 3333 ) shown schematically. The 64 fibers are on the array side ( 22 ) a distance of 4 mm from each other. The core and the cladding of each fiber are square and have a side length of 100 microns and 120 microns. On the front side ( 3333 ), the fibers are arranged in a line, wherein between each two light-guiding fibers ( 2222 ) always an intermediate fiber ( 4444 ) lies. (see 4 )

The intermediate fibers are square and the side length of the shell is 120 microns.

About the lenses ( 17 ) and ( 21 ), the sample plane is mapped to the fiber array at 4x magnification (see 1 ). Since in this example the number of fibers of the fiber bundle is less than the number of wells of a typical microtiter plate ( 18 ), the sample can additionally be moved via automatically controlled translation tables.

The advantage of this high-throughput screening system is that the use of a fiber bundle and special illumination ensures that all spectra of several wells of a microtiter plate can be measured simultaneously and quickly in the wells.

Characterized in that the fibers of the bundle are arranged on the side facing the specimen in a line and on the side facing the sample fibers are arranged in an array in two dimensions, so that between the fibers each have a certain, the same distance for all fibers, and the illumination is based on the multiple imaging of the end face of a highly multimode optical fiber through a microlens array, this simultaneous readout of the cavities is enabled.

• • Alle Spektren werden gleichzeitig ohne spektrale und ohne laterale Rasterung aufgenommen,
• • die Anregung erfolgt mit depolarisierten und inkohärenten Licht,
• • die Probenbeleuchtung für eine maximierte Anregungsleistung wird gewährleistet,
• • ein quantitativer und qualitativer Vergleich der Proben in den Kavitäten ist möglich
• • die Aufnahme aller Spektren ist sehr schnell und
• • eine Maximierung der übertragenden Strahlung wird gewährleistet,

so dass dieses System alle erforderlichen Anforderungen zur Hochdurchsatzmessung in der Spektroskopie erfüllt.The high-throughput screening system thus has the following advantages:

• • All spectra are recorded simultaneously without spectral and without lateral screening,
• Excitation occurs with depolarized and incoherent light,
• • sample illumination for maximized excitation power is ensured
• • a quantitative and qualitative comparison of the samples in the cavities is possible
• • the recording of all spectra is very fast and fast
• • Maximizing the transmitted radiation is ensured

so that this system meets all the requirements for high-throughput measurement in spectroscopy.

All features illustrated in the description, the following exemplary embodiments, the claims and drawings can be essential to the invention both individually and in any desired combination.

LIST OF REFERENCE NUMBERS

1

laser

2

telescope

3

pinhole

4

lens

5; 333

Front side (laser-facing side)

6

illumination fiber

7

vibrator

8th

Front side (side facing away from the laser)

9

lens

10

Microlens array

11

between level

12

Objective lens of the infinite microscope

13

cover

14

line filter

15

rotatable linear polarization filter

16

filter

17

Lens of the infinite microscope

18; 222

microtiter plate

19

cut-off filter

20

linear polarization filter

21

lens

22

Fiber array

23

fiber bundles

24; 3333

Front side (spectrometer facing side)

25

spectrometer

111

Walls of the microtiter plate

444

Fibers of the fiber bundle

1111

Front side (side facing away from the spectrometer)

2222

light-conducting fiber

4444

between fiber

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• WO 2013/064159 A1 [0005]
• DE 10237394 A1 [0005]
• EP 1463392 A1 [0006]
• EP 1651934 A1 [0006]
• EP 1766350 A1 [0006]
• EP 1766351 A1 [0006]
• WO 2012/165837 A3 [0006, 0006]

Claims (11)

High-throughput screening system for performing optical measurements in spectroscopy, comprising an excitation unit with a laser ( 1 ) and an optical route, a readout unit with a fiber bundle ( 23 ) of optical fibers and a spectrometer ( 25 ), a microtiter plate ( 18 ) with cavities for individual samples and a movement and positioning unit for the arrangement of the wells of the microtiter plate ( 18 ) relative to the fiber bundle ( 23 ), characterized in that the fiber bundle ( 23 ) is a dimension-reducing fiber bundle, which on its side facing the sample ( 22 ) a rectangular or square arrangement of the optical fibers and on its side facing the spectrometer ( 24 ) has a line-shaped arrangement of the optical fibers, and that an optical highly multimode illumination fiber ( 6 ) via a lens and a microlens array ( 10 ) in an intermediate image plane ( 11 ) multiplied and then over two more lenses ( 12 ; 17 ) can be imaged in the plane of the sample, wherein all cavities can be simultaneously illuminated and measured by the excitation and read-out unit with a radiation in which the producible radiation to be measured is transmitted via a system of optical lenses ( 17 ; 21 ) from the cavities to the fiber bundle ( 23 ) and from the side facing the spectrometer ( 24 ) of the fiber bundle ( 23 ) into the spectrometer ( 25 ) is transferable. High-throughput screening system according to claim 1, characterized in that the illumination fiber ( 6 ) a high length in the range of several decameters and a vibration device ( 7 ), so that the polarization of the laser ( 1 ) emissive radiation in the illumination fiber ( 6 ) by the vibration device ( 7 ) and the length of the illumination fiber ( 6 ) is destructible. High-throughput screening system according to claim 1, characterized in that the optical path from the laser ( 1 ) over the illumination fiber ( 6 ) and a line filter ( 14 ), so that in the illumination fiber ( 6 ) is removable. High-throughput screening system according to claim 1, characterized in that the fiber bundle ( 23 ) of optical fibers ( 2222 ) having a round or angular cross-section, between which non-conductive intermediate fibers ( 4444 ) are square or round cross-section, which the light-conducting fibers ( 2222 ) on the spectrometer ( 25 ) facing side ( 24 ; 3333 ) spaced from each other. High-throughput screening system according to claim 1, characterized in that the light-conducting fibers ( 2222 ) on the sample-facing side are arranged at a distance from each other in the array, so that only the light which is generated in the samples, and not that from the walls of the microtiter plate ( 18 ) outgoing light is receivable. Method using a high-throughput screening system according to one or more of the preceding claims 1 to 5, in which the samples in the wells of the microtiter plate ( 18 ) simultaneously with uniformly unpolarized light or by using an additional linear polarizing filter ( 15 ) are irradiated with linearly polarized light, in which the laser ( 1 ) generates a monochromatic radiation which is transmitted via the illumination fiber ( 6 ) as well as the line filter ( 14 ), so that in the illumination fiber ( 6 ) the scattered radiation is removed, wherein simultaneously with the vibration device ( 7 ) on the illumination fiber ( 6 ) a vibration is generated so that the polarization and the coherence of the laser ( 1 ) emissive radiation in the illumination fiber ( 6 ) by the vibration device ( 7 ) and due to the length of the illumination fiber ( 6 ) is destroyed, and the excitation light thus generated is imaged onto the samples so that the illuminated surfaces after imaging on the fiber bundle ( 23 ) in their extent with the fibers ( 444 ) of the fiber bundle ( 23 ), so that homogeneous and intensity-identical illumination surfaces in the sample-containing wells of the microtiter plate ( 18 ), the radiation emanating from the samples with the fiber bundle ( 23 ) and then a computer-aided standardization of the data read out with a homogeneously scattering sample in order to obtain accurate quantitative statements with respect to the intensities of the scattered radiation emanating from the samples. A method according to claim 6, characterized in that the light before entering the illumination fiber ( 6 ) and through a pinhole ( 3 ) is controlled in performance. A method according to claim 6, characterized in that the microtiter plate ( 18 ) is moved during the irradiation so that each cavity illuminates a larger sample area and thus a higher maximum possible illumination power is enabled, whereby a shorter necessary exposure time or a higher ratio of signal and noise is possible. Method according to claim 6, characterized in that by replacing the lens ( 9 ) between the illumination fiber and the microtiter plate ( 18 ) the high-throughput screening system very quickly and automatically to different microtiter plates ( 18 ) can be adjusted without the functionality of the lighting changing. Method according to claim 6, characterized in that several lasers ( 1 ) into the illumination fiber ( 6 ) can be coupled in order to do so only by changing the filters ( 14 ; 16 ), with otherwise constant illumination, to be able to perform measurements at different excitation wavelengths, or that several lasers ( 1 ) simultaneously into the illumination fiber ( 6 ), so that the sample is excited by these light sources at the same time, and the radiation emanating from the sample is examined, or that the fiber-based sample illumination is carried out not by the incident light method, but by another lens, or in the transmitted light method. Use of the method according to one or more of the preceding claims 6, to 10 for the simultaneous recording of at least two spectra for calculating difference spectra based on the simultaneously measured reference spectrum, for the analysis of the smallest spectral shifts in optical spectroscopy.

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