EP1301924A2

EP1301924A2 - Device for carrying out biochemical fluorescence tests

Info

Publication number: EP1301924A2
Application number: EP01955276A
Authority: EP
Inventors: Wolfgang Karthe; Ralf WALDHÄUSL; Andreas BRÄUER; Norbert Danz; Ralf Kindervater
Original assignee: Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV; Trace Biotech AG
Current assignee: Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV; Trace Biotech AG
Priority date: 2000-07-19
Filing date: 2001-07-19
Publication date: 2003-04-16
Also published as: WO2002006836A3; AU2001277486A1; US20040021867A1; WO2002006836A2

Abstract

The invention relates to a device for carrying out biochemical fluorescence tests with which the different biochemical interactions can be detected. The aim of the invention is to be able to economically detect a very large number of individual samples with a high level of sensitivity and to obtain a high local resolution. To these ends, the invention enlists the use of a device with which linearly polarized light of a laser diode is directed onto a plate-shaped support via an optical configuration comprised of at least one polarization beam splitter, of a μ/4 plate and of a focussing optical element. In addition to binary optically detectable information structures, a multitude of fluorophore-marked samples are also discretely arranged on the support that rotates around an axis. Light reflected on the information structures is directed onto an optical detector via the optical configuration in order to detect the information, and fluorescent light emitted by the fluorophore-marked samples is directed onto an optical detector for the fluorescent light via a spectral filter that separates in a wavelength-selective and spatial manner.

Description

Device for carrying out biochemical fluorescence tests

The invention relates to a device for performing biochemical pluorescence tests, with which the different biochemical interactions can be detected. Various so-called assay formats known per se, for example fluorescence immunoassays and tests for decoding the genome of plants or animals, can be carried out. The invention can be carried out very particularly advantageously for the examination of a very large number of samples in a short time, as is desired in the so-called “screening applications”.

In the known prior art, the use of rotating carrier media for a relatively large number of samples is proposed for this purpose, and the evaluation and implementation of the examinations is said to be with the help known technology and here in particular by means of CD or DVD technology.

Such proposed solutions are addressed in WO 98/12559 AI, WO 99/35499 AI and WO 00/26677 AI.

The content of WO 00/26677 AI relates essentially to the modification of known CD or DVD and their manufacturing process. There is an indication of the possibility of carrying out tests with fluorescence excitation and measurement of the excited fluorescence light. However, only solution approaches have been explicitly described in which colloidal particles, for example gold, are used with a partner of such a binding system for the detection of successful binding of at least two such partners, such as known receptor-ligand systems. As a result, the reflection and absorption behavior which occurs as a result of colloidal particles and which occurs on molecules bound in this way can be used and corresponding statements, if appropriate also in quantitative form, can be obtained by appropriate optical detection.

If, on the other hand, the fluorescence analysis technology frequently used in this field is used, the detection of the fluorescence light must be measured selectively in terms of wavelength with high sensitivity and in particular with very high spatial resolution, as is not readily possible optically with the known CD or DVD technology ,

However, the advantages of such systems, namely the high speed of signal acquisition and in particular the possibility of U> to CJ1 O Ul O in

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if circularly polarized light is again on the λ / 4 plate and is again converted into linearly polarized light, the plane of polarization of the reflected light being different from the plane of polarization of the light emitted by the laser diode

Is rotated 90 °. As a result, the reflected light can be deflected with the polarization beam splitter and directed onto the optical detector, so that a clear separation of the information signals obtained with the reflected light from the light emitted by the laser diode can be achieved.

To reduce the undesired influence of extraneous light, it is advantageous to arrange an additional optical filter between the spectral filter and the optical detector for the fluorescent light. A band or edge filter matched to the respective wavelength of the fluorescent light can be used for this.

In particular, when using a carrier which is completely or partially optically transparent at least in areas in which fluorophore-labeled samples are arranged, there is the possibility of the optical detector for the fluorescent light and the correspondingly required wave-selective and spatially separating spectral filter on the side of the carrier to be arranged opposite the side on which the laser diode and the optical arrangement are arranged.

In this case, however, the optical elements arranged on both sides of the carrier should be able to be moved synchronously, which can be achieved, for example, by a rigid mechanical coupling. In certain cases, however, it can also be expedient to arrange all the optical elements on one side of the carrier, so that they can be moved back and forth together along the radially outward axis. The spectral filter, with the aid of which the fluorescent light is directed at the optical detector for the fluorescent light, wavelength-selectively, can be integrated into the optical arrangement, so that the light emitted by the information structures from the carrier also hits this spectral filter, but from remains unaffected.

However, in addition to the laser diode, it is also possible to use at least one second light source that is as monochromatic as possible, which can also be a corresponding laser diode but also an LED. This light source only emits light for fluorescence excitation of one or more appropriately selected fluorophores. The light from this second light source can be directed via a wavelength-selective and spatially separating spectral filter (dichroic beam splitter) onto the carrier and consequently also onto the fluorophore-labeled samples. The optical elements of the optical arrangement, which are used to obtain the information signals from the information structure by appropriately superimposing the light from the laser diode and the second light source, can also be used.

With such an arrangement it is possible to carry out fluorescence tests with at least two different fluorophores, in which fluorescence with different wavelengths can be excited, if the first laser diode also emits light with a suitable wavelength. Since both the information structures, as well as the fluorophore-labeled samples can be arranged in different planes in or on the support, it is advantageous to change the focal length of the focusing optical element, which is then in the form of an objective with variable

The focal length can be designed to vary accordingly, so that the focus lies in the respectively desired plane and the desired information and in particular the fluorescence signals can be recorded with a very high spatial resolution.

With the device according to the invention, the detection of both the optical information from the information structures and the detection of the fluorescence signals can take place very confocally.

To ensure the desired high sensitivity, in particular for the fluorescent light, photo ultiplier tubes should be used as a suitable optical detector

(PMT), avalanche photodiodes or particularly sensitive photodiodes with preamplifiers can be used.

Additional collimators and condensers can advantageously be arranged in the beam path of the different types of light in order, depending on the need, to expand and parallel align them or to focus them, as is particularly desired for the light to be directed onto the optical detectors - pass.

Another possibility is not to direct the fluorescent light directly onto an optical detector for fluorescent light via spectral filters and filters, but rather to direct the fluorescent light into a suitable focusing lens 1 1 1 1 XI 0 0 rö ^ Φ 0 Φ XI -H υ 1 Φ Φ 1 1 = rö SH 1

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4-J τj 4-J w -Q Φ Λ τj 0 -H Φ -H T-i -H υ τi -H SH 0 0 Φ rö <-H P. Φ Φ = rö φ = 0 Φ 0

-H 0 ^ φ Φ 4-J N Φ I Φ 4J xi = rö XI φ. 0! Λ SH n M 01 0

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Xl X! 0 4-J 0 CQ -0 0 _^ Φ. rH φ φ 0 rH 4H 4-J CQ Φ XI Φ T3 Φ B

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zenzanalyse other optical variables that change as a result of occurring biochemical interactions, such as changes in reflection and absorption, can also be detected, so that the test spectrum can be expanded.

Such changing quantities can optionally be detected without additional changes to the device according to the invention with the optical detector, which anyway contains the information contained in the information structure of the carrier.

The invention will be described in more detail below on the basis of exemplary embodiments.

Show:

Figure 1 shows the schematic structure of an example of a device according to the invention;

Figure 2 shows a second example with additional collimators and condensers;

FIG. 3 shows a third example with an arrangement of optical elements that is different from the example according to FIG. 2;

Figure 4 shows another example compared to

Examples according to Figures 2 and 3 changed arrangement of the optical elements;

FIG. 5 shows an example with an additional light source for fluorescence excitation;

FIG. 6 shows an example of a device according to the invention with an optical fiber for fluo- reszenzlichtführung;

FIG. 7 shows an example of a device according to the invention with separate optics for fluorescence excitation and detection;

FIG. 8 shows an example of a carrier that can be used in a device according to the invention;

Figure 9 shows another example of such a carrier;

Figure 10 shows an example of a carrier;

Figure 11 shows an example of a composite beam;

Figure 12 shows another example of a composite beam;

FIG. 13 shows an example of a composite carrier with information structures arranged in two levels;

FIG. 14 shows another example of a composite carrier with information structures arranged in two levels;

FIG. 15 shows a further example of a carrier with two information structures arranged on different levels;

FIG. 16 shows an example of a composite carrier with an information structure in one level; FIG. 17 shows a further example of a composite carrier with an information structure arranged in one plane;

FIG. 18 shows a structure in a highly schematic form, as can be used in an example according to FIG. 7, and

Figure 19 shows the basic structure of a device according to the invention with an additional dispensing device.

In devices such as those shown in FIGS. 1 to 7, laser diodes 21 or other light sources 29 can be used, the light of which has wavelengths with which fluorescence per se known fluorophores can be excited. Preferred wavelengths are e.g. 635 nm, 650 nm and 780 nm, with laser diodes 21 already available for this.

As shown in FIGS. 1 to 6, an optical arrangement A can be used in a device according to the invention, with which linearly polarized light of a laser diode 21 can be focused on or also in a plate-shaped carrier 1.

The light of the laser diode 21, which laterally, radially with respect to the axis of rotation of the carrier 1 (not shown), can of course be moved back and forth together with the optical arrangement A, so that in connection with the rotation of the carrier 1 the whole Carrier surface can be scanned.

The linearly polarized light from the laser diode 21 is transmitted through a polarization beam splitter 22, showed an example of a double prism, wherein the one base surface of a prism can additionally be provided with a λ long pass coating. The λ long pass coating taking into account the wavelength of the laser diode 21 and / or

Light sources 29 or the arrangement of the polarization beam splitter 22 in the optical structure may be required.

In this example, a wavelength-selective and spatially separating beam splitter 26 is arranged below, the function of which will be discussed below. A λ / 4 plate 23 is arranged thereafter, with which the linearly polarized light is converted into circularly polarized light.

A λ / 4 plate 23 is followed by a focusing optical element 24 with which the light can be focused on the surface of the carrier 1 or into the interior of the carrier 1. The position of this focusing element 24 can advantageously be changed, as indicated by the double arrow drawn in the vertical direction, so that the focus position can be changed. This makes it possible for light to be focused as required on a plane in which an information structure 3, 4 or a fluorophore-labeled sample is arranged.

The light reflected by the information structure 3, 4 by means of so-called “pits or lands” formed there is the carrier of binary information which can be digitally recorded and processed in an electronic evaluation and control unit.

That reflected by the information structure 3, 4

Light then arrives again via the focusing op- table element 24 to the λ / 4 plate 23, where it is linearly polarized again. The plane of polarization of the reflected light is rotated by 90 ° with respect to the emitted light linearly polarized by the laser diode 21. By changing the

Polarization plane, it is possible to separate the reflected light via the polarization beam splitter 22 and, as can be clearly seen in FIG. 1, direct it onto the optical detector 25, which is preferably a quadrant diode.

If fluorescence in a pre-marked sample is excited with the light of the laser diode 21, the emitted fluorescence light passes through the focusing optical element 21, the λ / 4 plate 23, to the spectral filter 26, with which a spatial separation of the fluorescence light is also to be achieved. The spectral filter 26 is also shown here as a double prism and a dichroic beam should preferably be used for this purpose in order to separate the fluorescent light and to direct it onto the optical detector 27 for the fluorescent light. The fluorescent light, since it is not polarized, remains unaffected by the λ / 4 plate 23.

To suppress the influence of extraneous light, an additional filter 28 is arranged in front of the optical detector 27 for the fluorescent light, so that the signal-to-noise ratio can be improved.

The example of a device according to the invention shown in FIG. 2 differs from the example according to FIG. 1 only in the additional use of a collimator 32 and additional condensers 33, the latter focusing the light on the optical detectors 25 and 27. In the example shown in FIG. 3, only the polarization beam splitter 22 and the spectral filter 26 and accordingly also the optical detectors 25 and 27 are interchanged with respect to the laser diode 21.

The example according to FIG. 4 is intended to clarify that the light guidance of the light from the laser diode 21 can take place in a different form. It will

Light of the laser diode 21 is first emitted parallel to the surface of the carrier 1 and can be deflected by means of the spectral filter 26 by 90 ° in the direction of the carrier 1. The spectral filter 26 is then provided with a non-polarized λ long pass coating.

With such an arrangement of the optical elements, the available space inside a device can possibly be better used.

FIG. 5 shows an example of a device according to the invention in which an additional light source 29, which, as already mentioned in the general part of the description, can also be a suitable laser diode, is present. However, the light source 29 should emit light with wavelengths that differ from the light of the laser diode 21.

At least the light from the laser diode 21 or the light source 29 should be able to excite fluorescence from a fluorophore, but advantageously both light sources 21 and 29 can excite fluorescence from a fluorophore separately.

Is light with two fluorescence-stimulating wavelengths gene used, should not be shown here, a second optical detector 27 ^λ for fluorescent light and an additional light with different fluorescent light ^wavelengths spatially separating element should be used.

A solution for this can be seen in FIG. 6. In this example there is an optical fiber 31 with the additional spectral filter 26 and the two optical detectors 27 and 27 ^x .

In the example as shown specifically in FIG. 6, however, a second light source 29 has been dispensed with. However, in order to nevertheless detect fluorescent light with different wavelengths, different fluorophores that can be excited with approximately the same wavelength but emit with different wavelengths can be used. The fluorescent light is coupled into the optical fiber 31 via the condenser 33 and decoupled by means of the collector 32 and directed onto the wavelength-specific and spatially separating spectral divider 26 ', with which the fluorescent light of different wavelengths is directed in a separate form at the two optical detectors 27 and ^27λ can be.

In the example shown in FIG. 7, the binary, optically detectable information of an information structure 4, which is arranged within the carrier 1, is by means of a laser diode 21, a polarization beam splitter 22, the λ / 4 plate 23 and the focusing optical element 24 and the optical detector 25 and can use the already mentioned evaluation and control electronics to control the movement (tracking) and on the other hand to the local one Assignment of fluorescent signals originating from fluorophore-labeled samples can be used.

A second optical system, which is used exclusively for fluorescence analysis, is arranged on the opposite side of the carrier 1. In this device, a light source 29, the light of which can excite fluorescence from a fluorophore, is again directed onto a spectral filter, which is designed here as a dichroic beam 30, and from there via a further focusing optical element 24 ′ onto fluorophore-labeled samples, which are here are arranged within a surface structure which is formed on the carrier 1. The emitted fluorescent light passes through the focusing optical element 24 ^λ through the dichroic beam splitter 30, an optical filter 28 onto the optical detector 27 for the fluorescent light. As indicated schematically in FIG. 18, the two optical parts arranged above and below the carrier 1 can be mechanically rigidly connected to one another and consequently moved synchronously.

However, it becomes a suitable one for fluorescence excitation

Laser diode 21 and an at least partially transparent carrier 1 used, the additional light source 29 and the dichroic beam splitter 30 can optionally be omitted in the example shown in FIG. For this purpose, for example, the information structure 4 can be interrupted in areas in which fluorophore-labeled samples are arranged, so that the light can reach the sample.

However, there is also the possibility of

r-- ε Φ I xi τj 0 u

SH 0 -H

Q 0 -H ro CQ τj

SH Φ a. 4J φ rA

CQ. di

-H 4-J = ro 00

Φ SH

0 0 EH SH

Φ xi 0

SH J CQ tn

0 -H φ -rH

01 Φ 0 tϊi

-ri l -H

Xi φ Xi tn ü

0 0 rA ro

Φ -H Φ 0 τj Φ -H

0 for rH o Φ 00 CQ Φ tn -H -H rH 4-> Φ Pi

0 XI PQ CQ

4H υ -H

XI -H Φ Φ ü rH 4-J PQ rö SH Dl

0 Φ -H Q Φ 0 ε

0 ro! > φ Xi Φ τj Di xi

Φ ü τj 4-> 0i -H

0 SH CQ

0 Φ SH

-H 0 4-y

00 Q Di Φ

CQ -H τj

0 PH -H

0 X Φ tn 0 0 Xl

-H 4H -H ü f, CQ v ^ CQ CQ SH f »i 0 rö rö Φ

H τj P 4-J

O

ui o L0 O o LΠ rH m

SH 0 1 Φ r ~ - ^ 0 CQ H X ε CQ 1 1 *. SH Xl

CQ X CQ Φ O SH τj H 0 0 xl 0 Φ 0 xi = rö -0 ε CQ o Φ Φ CQ 0 4-J tn SH H τj 4-J SH φ φ rö υ 0 4-J rö o SH -ri 0 -ri rH 1 ω τj τj Φ SH SH = rö Φ 0 CQ Φ 0 0 SH 4-> -ri -H rH CQ X --rö EH Φ M Φ CQ

Q 4-J 4-J O SH τj 0 -ri CQ τj -ri 0 Φ 0 τj J 4-J rH Φ rH 4-J -ri 0

H Φ tn 0 CQ EH 0 Φ CQ 0 φ φ Φ Di 0 0 rö O 4-> 0 0 4-J xl Φ fr rö U Di 0 Φ CQ 0 rö 4-JO Sä Sä = rö φ 0 D ε CQ SH Φ Φ -ri υ α. ö ö SH ε l r 0 ε 0 r 0 = rö 4-J -ri 4J SH SH φ τj 0 4-J r H -r ε H rö i φ r SH SH rH ε -i

0 0 -i CQ l -ri o rH SH Φ 0 0 Φ -ri 4-J Φ 4-J 0 Φ 0 r Φ rH

CQ Φ rH -ri φ 0 Φ -ri τj rö Φ τj. CQ -ri rH 4H -ri 4J Φ P, rA CM l PQ

0 τj CQ 4-> Dl -ri 4-J rH ε 0 0 rH 0 H 4-J -ri 0 Φ X Sä φ 4-J rA 0

X 0 rö φ -ri rö -ri SH XI 0 rö φ 0 X Φ H CQ Φ 0 4-J Φ Φ Φ CQ Φ

0 = ro 0 ε SH ε X, X! 0 CJ ^ rö τj 4-J φ 4-J rA 4-J Φ CQ 4-> τj tn rö 4-J

P-, SH Φ in XI rH Φ 4H -ri X _* . X rA 0 0 τj 4H CQ 0 φ rö -ri φ τj = rö

Φ XI 0 Φ Φ 0 φ Dl 0 CQ 4-J ε 0 0 4H Φ 0 Φ CQ 0 tn SH φ -ri 4->

SH> υ 4H CQ -ri -ri -ri CQ H Xl 0 SH -ri SH Φ 4-J Φ SH rö -ri SH 4-J Xl for τj -ri

Φ CQ 0 Pl τj 0 CQ ü rH 0 4-J SH Φ SH 4H 4-> rö CQ CJ CQ 0 τj Φ -ri H Φ CQ CM rö Φ Φ -ri Φ Pm 0 CQ φ τj Φ τj SH rö τj X ! • CQ 0 ro

0 4-J rH -ri -ri 0 xi τj Φ CQ Φ CQ rH -ri 0 Φ i ε 0 0 SH φ «

4 Xl P, SH rö • Φ Φ φ CM -r 0 CJ 0 0 4-J 0 CQ CQ -ri 4-) φ SH r-- CQ Φ φ 0 CM

-ri 0 0 Φ 0 0 PQ 4-> X! Φ 0 ß rö Φ 0 0 -ri -ri X! X xi LD 0 H τj 4-J -ri φ

Φ τj tn Φ rö 0 0 rö xl 4-J 0 -ri φ Sä Φ φ υ CM 4H 0 0 0 φ Φ -ri

X τj 0 -ri 0 0 SH Sä Φ CQ X Φ rH 4-J Sä Φ tn rA φ 0 CQ Φ 0 0 4J τj

SH 0 φ CQ CQ 0 φ 4-J 4-J rH τj ro rH 0 rö rA tn CQ 4H SH SH H -ri τJ X! XI rö rö 0 τj 0 IS! = 0 4-> CQ -. rö -ri -ri 0 Φ O B -ri 0 Φ fr 0 X) -ri SH ^ CJ CQ

Xi 0 ro 0 X tn X! 4-J SH Φ φ φ 4-J 4-J> Φ 0 rö Ul 4-J 0 Φ Φ H SH 4-> CQ rH 4-J Φ Φ -ri 0 Φ 4-J Sä X! SH XI CQ 0 4J -ri Xl 4-> X ro ro X! > 0 CQ rö rH in SH 0 N 0 φ ß τj CQ Φ rö Φ ü Φ co 4H Φ CM CJ 0 Φ H SH τj Xi τj

Φ Φ φ Φ CQ Φ IM 0 X! -r— i P -ri -ri tn 0 CM O φ 4-J rH CQ r- 0 0

4-J tn -ri 0 Φ τj φ -ri φ 0 4-J XI SH 4-J XI 4-J Xl φ Xl 0 rH tn ro ß -

CQ 0 CQ 0 SH SH Dl Φ Sä ß 4J. X CJ rö xi 4 ü rö 4-J P rö Φ -ri 4-J -ri rA XI

0 -ri CQ -ri 0 Φ Sä SH CQ 4-J Φ ß τj u Φ rö 0 SH 0 SH N SH φ XI Ct, 0 CJ

-ri SH 0 4-> 0 Sä r ISI Φ 0 -ri Φ rA -ri -ri SH rö 4J 0 0 0 0 0 £ u SH φ SH

PQ SH X rö rH H Φ τj 4H φ SH rH τj 4-> CQ rö τj Φ <: tn φ -ri <Ά 0 τj 0

C 0 C 4-J

4H S εH CQ ε τj 0 rH Φ τj Φ SH CQ CQ X! τj Xl -ri XI: CÖ tn -ri τj

Φ> SH 0 ε -ri Φ -ri SH 0 τj Φ -. X! CQ 0 0 ü φ 4-J C ü ε -ri φ rö

-ri 0 φ CQ 0 SH SH Φ 4-J Xi Φ C τj 0 rö ß Φ 0 CQ -ri CQ Φ CQ Φ PH Ά τj τj 00 CQ 4H -ri rö Dl Φ O Xi rö φ Φ SH 0 rö SH ß τj -ri -ri τj CQ 0 -ri SH oil

Φ 0 τj 4H -ri τj CM SH tn CQ φ -ri 10 -ri 0 τj 4-1 = rö φ τj Φ X! SH LD

0 CQ τj H SH PL, φ 0 4 CQ -ri -ri 5 0 0 φ τj Pi Xl rH τj rH rH υ Φ rH rö rH xl Φ -ri SH Φ CQ 0 Φ 4-J Φ Φ Φ CQ SH -. 0 0 J τj 4-> Φ rö τj

Xl φ Φ CJ 0 Φ τj X! rö Sä X τj 4-J 4-J 4J φ ro 0 -ri 0 0 0 4-J -ri 0 SH tn rö 4-J -ri 0 4J -ri 0 0 rH Φ SH SH H Φ - X! ε CQ H -ri -ri -ri P, tn 0

0 in -ri τj ro CQ 0 Φ 0 Cß ^• Φ -ri rA 0 Φ Φ 0 4-> 0 SH 0 0 Φ CQ ε CQ rA 0 oil

0 4-J Φ = 0 ε Φ τj Φ Φ 4H Sä -ri -ri τj Φ 0 Φ> CQ <0 SH -ri Φ 0 -ri

SH CQ rH Φ rH Φ τj 0 XI 0 »4-> Φ CQ CQ rH τj ε 4-> oil CQ = 0 Φ H φ φ -ri 0 Pc

Φ SH 0 XI -ri Φ τj H 0 CJ φ cn SH -ri CQ CQ 0 rH -ri X 0 4-J rö SH τj> PQ fr τj τj Φ 0 0 Sä Dl φ X! CQ τj Φ Xl φ 0 0 Φ -ri 0 rö Xl τj tn 0 4-> Ul XI Q Φ Sä 4H -ri Sä SH -ri -ri 0 ü X! X X tn X! SH rH ü SH -ri SH 4H CQ -ri 0 U

C5 0 ro SH 0 -. 0 Φ Φ £ Φ 0 -ri 0 ro 0 0 0 0 Φ 0 4-J Φ -ri 0 Φ Φ Φ ro ro φ 0 ro

4H PQ CQ ro ro PQ H i IS! X! 4-J Φ XI P 4H 4H red N CQ oil, 0 CQ> PQ Dl K P PQ! 0

O

mtlo tn rH CM ro ro

are finally arranged within the substrate 2 arranged there above.

The examples according to FIGS. 16 and 17 again use only a single information structure 3, 4 which is formed within the substrate 2 arranged above and only differ in the arrangement of the cavities 10 in the examples shown in FIGS. 16 and 17.

In the examples for carrier 1, as shown in FIGS. 13 to 17, there are no pauses in the acquisition of information signals, which can be obtained with the aid of information structures 3, 4, if at the same time fluorescence signals by corresponding fluorescence excitation are detected by fluorophores.

FIG. 19 is intended to indicate a possibility in a schematic form that enables high-level automation of sample preparation and sample evaluation.

For this purpose, examples of a device according to the invention, as shown in FIGS. 1 to 6, can be used below the carrier 1. A dispensing device for samples is arranged above the carrier 1 and can be controlled with the aid of the information signals obtained, so that the sample can be applied with high precision with regard to the respective position and volume.

In the biochemical preparation of supports and samples, the knowledge known per se can be readily used, so that the most varied of biochemical interactions can be aims and can be detected with the solution according to the invention.

Claims

claims

1. Device for carrying out biochemical fluorescence tests, in which linearly polarized light from a laser diode (21) consists of an optical arrangement (A.) Consisting of at least one polarization beam splitter (22), a λ / 4 plate (23) and a focusing optical element (24) ) is directed onto a plate-shaped carrier (1),

the carrier (1) rotating about an axis is provided with binary, optically detectable information structures (3, 4) and a plurality of fluorophore-labeled samples are discretely arranged on the surface and / or inside the carrier (1);

light reflected from the information structures (3, 4) is directed through the optical arrangement (A) for detecting the information onto an optical detector (25) and

Fluorescent light emitted by fluorophore-labeled samples is directed via a wavelength-selective and spatially separating spectral filter (26) onto an optical detector (27) for the fluorescent light.

2. Device according to claim 1, characterized in that the spectral filter

(26) is a dichroic beam splitter provided with a λ short-pass coating.

3. Device according to claim 1 or 2, characterized in that the spectral filter (26) or the polarization beam splitter (22) is provided with a λ long pass coating.

4. Device according to one of claims 1 to 3, characterized in that between the spectral filter (26) and the optical detector (27) for the

Fluorescent light an optical filter (28) is arranged.

5. Device according to one of claims 1 to 4, characterized in that the spectral filter (26) and the optical detector (27) for the

Fluorescent light is arranged on the side of the carrier (1) opposite the optical arrangement (A).

6. Device according to one of claims 1 to 5, characterized in that the spectral filter

(26) is an integral part of the optical arrangement (A).

7. Device according to one of claims 1 to 6, characterized in that a second light source (29) for fluorescence excitation is present;

the light of this light source (29) is directed onto the carrier (1) by means of a second dichroic beam splitter (30), the

Superimpose light rays from the laser diode (21) and the light source (29).

8. Device according to one of claims 1 to 7, characterized in that the focal length of the focusing element (24) is variable.

9. Device according to one of claims 1 to 8, characterized in that the detection of ,

4-J

CQ

ri

SH ro

Xi

H

Φ

4-J CQ

o rH ro rH H rH rH

90

SO

©

© o

1 φ SH 1

I SH 0 Φ 1 1 1 0 SH Q 0

CQ SH xi. -. = rö n XI CQ rH o Xl CJ SH ^• o -ri XI Φ SH 4-J rö Φ P rö rö H -ri Φ 0 <* SH CQ υ -ri - CJ rö Φ - _\ BX υ 4-> EH XI NU ε 0 α. Xi 0 -ri oo EH 0 rö Xi Φ CQ 0 tn »τj Φ SH SH φ ü 0 CQ 0 SH -ri

-ri φ - ro SH SH 4-J -ri = ro ro 0 4-J ro 0 τj CQ -ri Φ Φ φ 0 τj

H Φ SH 0 i rH φ SH ε Di SH - - 0 CQ ε P Φ SH XI. SH 4J 4H SH

Φ Oil Φ _^ - _^ Φ tn Φ Φ 0 H CQ SH xl τj 0 0 0 O SH 0 0

Φ CQ 0 0 τj H Dl Φ = rö -ri XI 0 0 φ rö 0 SH o -ri SH Φ 0 Φ H 0 xl rH Φ 0 -— '0 Xl SH N υ 4-> Φ Q, -, Φ pi , τj .. rH -ri X υ Φ 4-J 4-J 4H -ir CJ EH -ri ε Φ 4-> 4H XI o Xl -ri SH CQ SH Pl τj -ri ^• Φ SH SH r Cd X 0 CQ

= 0 4 Cß XI 0 H Φ = 0 4H -ri υ 0 = 0 φ O 4J Φ 0 Φ rH SH Φ 4-1

SH 4J υ rö Φ SH SH -ri Xi -ri. 4-Y rH SH SH 0, -. -ri O Sä Φ rö τj SH

Pl -ri τj -ri Di Φ Pl Φ τj £ X Φ 0 0 0 0 00 CQ> SH -ri ε OQ ε 0 rl 4-J = rö rH CQ τj 0 0 N 0 Dl 0 0> - 0 4J Φ τj SH 0

0 0 0 CQ SH = rö 0 ε 0 0 X! SH 4H> rH Φ Φ Φ N τj 0 0 0! CQ -ri = 0 EH 0 <Q t>> 4-J 0 4H CQ -ri 0 P, X! 4-> 0 CQ Xl> Φ

CQ 1 Φ rH rö CQ 0 _^ 4H CQ rö n. CQ τj φ CQ rö 0 CQ Pl SH

SH rö Φ M φ ε W SH rö Φ tn tn 0 CQ CQ 0 SH D rö SH -ri tn 0 SD rö o SH rö

Φ τj 4-J Φ oil -ri Φ τj X! 0 Φ H rö 0 4J 0 Φ H τj CQ 0 P 4H Φ H τj SH Φ X! τj SH -ri 4H SH τj 0 0 0 0 SH 0 0. 4Y 4Y 0X! O τj CD

- Φ CQ 0 0 Φ _^ SH SH -ri CQ 0 -ri 0 τj -ri τj XI CQ X Xl »0 CQ

CM ε 4J 0 rö -ri τj ε 4-J P xl Φ -ri 0 4-J SH Φ SH υ 4-> Φ 0 φ. -. J 4J rH Di rö

Φ Φ CQ Φ CQ 0 φ Φ = 0 X)> rö Φ 0 -ri 0 Φ 4J SH 0 φ H 0 Φ 4H 0 4H

0 0 0 Pl 4-> SH 0 0 0 4H 4-J ε 0 0 CQ ä SH 0 N 4-J -ri 4-1 ^ - ^ SH 0 0 SH

-ri XI: CQ SH Φ. -. -ri xi 0 Xl -ri rA SH SH -ri tq rH P, Xl 0 CQ φ Φ Pl l 0 tn Φ

Φ ü -ri O τj o Φ ü> υ ε Φ O Φ -ri 4-> CQ CJ Φ CQ SH SH CQ ü Φ -ri

-ri Φ P 0 -ri SH Φ τj 4H Φ φ SH 0 -ri N 0 tn X Φ 0 -ri 0 4->, -.

Xl Φ xi 0 xi Φ Φ 0 CQ XI 0 0 SH £ Xl Φ CQ 0 0 Ul oil Φ rH Xl <φ υ IS! o 0 Φ 4J o IS! BP 4-> J CQ H Φ rö φ = 0 N Φ -ri 0 -ri = rö N Φ υ rö 0 CQ Φ X! l 0 ro 0 XI CQ = 0 rH rH X! -r— 1 4H XI 0 SH 4-J SH τj SH Xl 0 N -ri _^

0 0 -ri 0 0 ü Φ 0 0 rö SH Φ SH φ 0 rö rH Φ ü 0 0 rö Φ b υ 0 0 CQ 00

Φ 0 Φ rö 4-J Φ 0 0 4 P, 4-1 Φ 0 Φ 4-J Dl rö Φ 0 ε 0 Φ rö Φ -rA X -

Di X 0 CQ SH = rö tn X 4H ISI N Q 4-J 4-> tn 4-J XI XI 0 X rH SH Φ -ri 0 0 X φ o

0 Φ SH CQ X! 4-J 0 Φ 0 0 0 -ri Φ -ri 4-J υ ü Φ C-, 0 4-J τj Φ Φ = 0 0

0 Di 4-J 0 0 Φ -ri 0 Di 0 Φ Ü ε τj ß -ri -ri 0 Dl 4H Cß τj 0 Di SH Φ

4-J X rH Φ tn t> 4-J φ N rH ε rH 0 Φ 0 0 H Φ φ Φ

XI XI Φ Xl 0 4H rö XI XI φ 4-1 SH CQ SH i 0 0 N τj SH Xl 0 H SH = 0 J Ü rA ü rH 0 «υ υ -ri CQ Xl Φ Φ φ ε -r 0 SH XI τj PQ 0

Xi φ 0 0 XI ü 0 4H -ri Xl υ 4->

-ri SH Φ CQ Φ rö -ri SH τj -ri rö SH τj τj φ 4-J Φ φ rö SH 0 N rö SH CQ SH ^

SH 0 Φ N φ SH 0 4H 0 tn CQ Φ ISI X! 4H 0 oil Φ _^^ • 4H 0 4-J Φ 0

SH τj Φ n 0 -P 0 SH τj MP 0 ε -ri CQ CQ 0 CQ 0 SH τj 0 SH _^ - _^ ^ S SH τj CQ 4-J SH

0 rö -ri 0 -ri H Φ Φ 0 rö = 0 Φ rH φ φ 0 rö -ri Φ SH Φ rö 0 rö H oo IS! Φ rö Φ 0 4-J

> τj τj rö Φ τj τj> τj 4H P> Cr-, 0 X! ro 4H Φ SH PM> τj SH Xi Xi> τj 4J 0 CQ

90 ^ tn VQ r oo

H rH H H H

©

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tn o tn o tn o rA H CM CM 00

and / or information associated with a fluorophore-labeled sample.

19. The method according to any one of claims 16 to 18, characterized in that the focal length of the focusing optical element (24) is set with the electronic evaluation and control unit so that light for exciting fluorescence of the laser diode (21) and / or the light source (29) is focused on a fluorophore-arked sample.

20. The method according to any one of claims 16 to 19, characterized in that fluorescent light emitted by fluorophore-labeled samples is separated from the fluorescence-exciting light by means of a wavelength-selective and spatially separating spectral filter (26) and directed to an optical detector (27) for the fluorescent light.