WO1999057310A2 - Instrument for analysis and diagnostics - Google Patents

Instrument for analysis and diagnostics Download PDF

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Publication number
WO1999057310A2
WO1999057310A2 PCT/EP1999/002918 EP9902918W WO9957310A2 WO 1999057310 A2 WO1999057310 A2 WO 1999057310A2 EP 9902918 W EP9902918 W EP 9902918W WO 9957310 A2 WO9957310 A2 WO 9957310A2
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WO
WIPO (PCT)
Prior art keywords
analysis chip
analysis
matrix
chip
biomolecule
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PCT/EP1999/002918
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German (de)
French (fr)
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WO1999057310A3 (en
Inventor
Hubert S. Bernauer
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Biochip Technologies Gmbh
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Publication date
Application filed by Biochip Technologies Gmbh filed Critical Biochip Technologies Gmbh
Priority to AU38252/99A priority Critical patent/AU3825299A/en
Publication of WO1999057310A2 publication Critical patent/WO1999057310A2/en
Publication of WO1999057310A3 publication Critical patent/WO1999057310A3/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Definitions

  • the invention relates to an analysis and diagnostic instrument for examining biological samples, in particular DNA molecules.
  • analysis chips A number of analytical and diagnostic instruments are known in the field of biological and medical laboratory technology. In certain configurations, these are usually also referred to as analysis chips. If the analysis chip is intended for the investigation of DNA molecules, it is called a DNA analysis chip, DNA array or DNA grid.
  • analysis chip and analysis and diagnostic instrument are used synonymously in the following.
  • Conventional analysis chips usually consist essentially of a carrier material, in the surface of which wells are etched, in which, in the case of a DNA analysis, various DNA molecules are incorporated.
  • hybridization By bringing a DNA sample to be examined into contact with the bound DNA molecules can lead to hybridization, which provide signals about the presence or absence and possibly the concentration of nucleic acids in the hybridization solution.
  • a special analyzer then reads out the hybridization data.
  • the detection of the hybridization reaction is conventionally carried out by optical methods, such as by coupling light through a grating. The mass accumulation disturbs the evanescent field, whereby a disturbance in the far field can be measured.
  • Conventional analysis chips generally use silicon as the carrier material, which contains a non-transparent medium on which the DNA molecules are bound in the wells.
  • Analog, not highly integrated systems, such as ELISA systems have in particular the disadvantages that the biological sample to be examined has to be applied individually to each reference molecule in the wells, the coupling of light to detect the hybridization reaction is inefficient, and an adjustment of the analysis chip in the analyzer is difficult , the administration and processing of data and parameters is complex and the costs for production and operation are very high.
  • the object of the invention is therefore to provide an improved analysis or diagnostic instrument or method.
  • the invention is based on the basic idea of providing an analysis chip for examining biological samples, which essentially consists of a carrier, on the surface of which at least one biomolecule matrix composed of individual dots on a bottom of a microfluidic structure, such as, for example a meander structure, or a planar surface is applied.
  • the biological sample to be examined is applied to one end of the microfluidic structure and flows along this structure to the other end, the sample flowing over the respective matrix points, each of which has an individual represent viduelle molecular species, triggers certain reactions.
  • this reaction is usually a hybridization reaction.
  • molecules can be detected in the sample which have a greater or lesser degree of similarity or homology with the complementary molecular species on the surface or a part thereof.
  • hybridization conditions depend, for example, on the temperature or the salt concentration in the sample and can be set by the person skilled in the art using customary methods. For example, the salt concentration in a sample can be adjusted.
  • the setting of hybridization conditions is known in the prior art and can e.g. based on the teaching in Harnes and Higgins, "Nucleic acid hybridization, a practical approach", IRL Press, Oxford 1985.
  • the molecule to be detected in the biological sample is a protein or peptide
  • a number of possibilities are available to the person skilled in the art for how he can detect this with the analysis chip according to the invention.
  • One possibility is the incorporation of the molecule into an antibody sandwich, the antibody (or a derivative thereof) that is not fixed to the surface preferably being marked in a detectable manner.
  • biological sample means in the broadest sense that the sample contains biological material such as nucleic acids or proteins or derivatives or fragments thereof.
  • nucleic acids can be modified by methods of recombinant DNA technology. adorned, fragmented, etc.
  • proteins may have been enriched from a natural source prior to analysis.
  • the sample is transferred from a natural source directly to the analysis.
  • the analysis chip also has a storage medium for easier data management and processing.
  • the analysis or diagnostic instrument has the advantages over the prior art in particular that the sample to be examined only has to be applied to the microfluidic structure at one point, as a result of which the process time can be considerably reduced and it can be produced in a simple and inexpensive manner , since the microfluidic structure can be produced simultaneously with the carrier, the theoretical signal strength can be increased by the microstructuring in the area of the samples, a more efficient reaction detection is possible, and / or production-related and / or other data, such as Configuration, production lot, zero control data, patient data, expiry date, analysis results and the like can be saved directly on the analysis chip.
  • Another advantage is that the results of the analysis can be determined with considerably greater certainty due to a matrix that is preferably at least double redundant.
  • the analysis is carried out with a high integration density and high parallelism. Only small volumes of the samples are required. Further advantages are the miniaturization of the analysis chip, the possibility of real-time and / or online evaluation at the same time as the hybridization reaction.
  • FIG. 1 shows a spatial image of an analysis chip according to the invention
  • FIG. 2 shows a section along the surface II -II of FIG. 1 to illustrate the coupling of light for the detection of fluorochrome molecules
  • 3 shows the detail III from FIG. II;
  • Fig. 4 shows the principle of light coupling for the detection of mass accumulation.
  • the analysis or diagnostic instrument 2 - in short analysis chip - shown in Fig. 1 consists essentially of a carrier or support frame 4, which in cross section e.g. has a U-shaped profile which is closed at its ends, so that a cavity 8 is provided under a surface 6.
  • a microfluidic structure e.g. a meander structure 10 is provided, which is embedded in the carrier 6 or preferably in at least one preferably transparent medium 12 and 14.
  • This transparent medium 12 and 14 is e.g. an insert made of glass, quartz glass, plastic, silicon, silicates.
  • the meander structure 10 is designed such that dots 16 of a biomolecule matrix can be applied therein, each point 16 of the matrix preferably representing an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.) At least one of the dots 16 is present on the analysis chip 2, namely on the medium 12, but preferably at least twice redundantly, ie also at least on the medium 14 or other media (not shown).
  • the surface of the analysis chip or the medium 12 and 14 can be planar (without a microfluidic structure) and to provide the at least one biomolecule matrix thereon.
  • a surface opposite the biomolecule matrix which contains the fluid volume with the sample to be examined, has the microfluidic structure.
  • the analysis chip 2 then becomes Analysis placed on this surface.
  • the microfluidic structure 10 can furthermore be provided both on the surface 6 of the analysis chip 2 and on the opposite surface.
  • the carrier 4 of the analysis chip is preferably a plastic, which is produced by micro injection molding.
  • Derivatives of polycarbonate are particularly suitable as carrier materials.
  • the covalent coupling of biomolecules is preferably carried out on semifluidic, gel-like polymer surfaces in the area of the samples (e.g. polymer brush).
  • the polymer surfaces have their own hydrodynamics, which gives the polymer surfaces special hydrodynamic properties and has a positive influence on the relationship between signal and background noise.
  • Semifluidic polymer surfaces are synthesized directly on the surface 6 of the analysis chip and can generally be used as nano- to microstructuring of surfaces. are referred to as polymer brushes.
  • the theoretical signal strength is enhanced by the semi-fluidic surface structuring in the area of the samples.
  • genetic material is isolated from a biological sample to be examined, specifically amplified and, if necessary, marked.
  • the probe material obtained in this way is applied to one end of the meander structure 10 of the analysis chip 2 by means of a device, for example a pipette or syringe.
  • the biological sample to be examined or the genetic material flows over the individual points 16 of the biomolecule matrix along the meandering structure 10, each of which represents an individual molecule species, and hybridizes in the case of at least partially complementary biomolecules.
  • a number of methods are available to the person skilled in the art for analyzing and evaluating the data of the analysis chip 2. Two basic methods are preferred according to the invention.
  • the evaluation can be carried out by a real-time measurement of the hybridization by detecting the mass accumulation by providing the analysis chip with a grating coupler, which causes a disturbance in the evanescent field.
  • the evaluation can be carried out by a real-time measurement of the hybridization by detecting the mass accumulation by providing the analysis chip with a grating coupler, which causes a disturbance in the evanescent field.
  • the analysis chip according to the invention shown in FIGS. 1 to 3 is particularly suitable for fluorochrome-marked systems.
  • the analysis chip shown in FIG. 4 is suitable for evaluation with a hybridization detection by mass accumulation.
  • the analysis chip 2 has adjustment means 18 on the outer edge of the carrier 4, e.g. Adjusting lugs or indentations, so that when inserted into an analyzer (not shown) for evaluating the examination is precisely adjustable.
  • adjustment means 18 on the outer edge of the carrier 4, e.g. Adjusting lugs or indentations, so that when inserted into an analyzer (not shown) for evaluating the examination is precisely adjustable.
  • the individual dots 16 of the biomolecule matrix as well as the loops of the meandering structure 10 are preferably themselves relatively close together, e.g. with a distance between 100 and 500 ⁇ m. It is ensured that the individual points 16 of the analysis chip 2 correspond to corresponding evaluation elements of the analyzer.
  • the analysis chip 2 also has at least one storage medium which can store operating data, production-relevant data such as configuration, production lot, zero control data, entered patient data, the expiry date, analysis results and other auxiliary data.
  • the storage medium is preferably an electronic one Memory chip 20, a magnetic strip 22 and / or a bar code 24, however, can also be any other known storage medium, such as ROM or RAM.
  • the analysis chip 2 shown in FIGS. 2 and 3 has, in addition to the features described with reference to FIG. 1, the features which are advantageous for the detection of fluorochrome molecules.
  • the analysis of a biological sample in the meander structure 10 of the analysis chip 2 takes place in that in an analyzer the analysis chip 2 from the side facing away from the meander structure 16, i.e. is illuminated by the cavity 8, which is shown schematically in the form of arrow 26.
  • the underside 28 of the surface 6 is preferably provided with an optical lens field 30, which enables confocal illumination.
  • the lens array 30 is preferably designed such that it is located exactly below the meandering structure 10, in particular exactly below the dots 16, so that the illumination of the individual dots 16 shown in FIG. 3 is made possible.
  • the lens field 30 and the carrier 4 of the analysis chip 2 can consist of a uniform material, which in one step e.g. can be produced by micro injection molding. However, the lens field 30 is preferably located on the underside 28 of the transparent medium 12 or 14 inserted into the carrier 4.
  • the illumination 26 from the underside 28 of the analysis chip 2 has the particular advantage that the intensity of the light source can be reduced since the light beams do not have to pass through a beam splitter.
  • the incident light 26 is focused by the lenses of the lens array 30 onto the individual points 16 of the biomolecule matrix, as a result of which the dots 16, which have caused a reaction with the sample to be examined, cause fluorescence through the fluorochrome molecules.
  • Other ways to prove the Molecules are based on the principle of bioluminescence, eg phosphorescence, radioactive labels, which can be detected by applying X-ray films.
  • a suitable evaluation circuit for example an optoelectronic circuit, can then be used to further process the data obtained with respect to the sample to be analyzed. This data can be further processed together with the data stored in the storage medium and / or written into the memory of the analysis chip 2.
  • FIG. 4 shows the coupling of light for analysis chips in which the hybridization detection takes place via the detection of the mass accumulation.
  • an optical method is used in which the light is coupled in via a large number of gratings 32, as a result of which the evanescent field is influenced. This influence can be measured in the far field 39.
  • Exactly one optical grating 32 is assigned to each point 16 of the biomolecule matrix in the microfluidic structure 10, so that the efficiency of the light coupling can be increased considerably.
  • the light 34 from an illumination source is focused on the grating 32 arranged in the matrix via a two-dimensional lens field 36 or via a two-dimensional Fresnel lens field, for example a hologram or diffractive optics.
  • the analysis data obtained can be further processed and / or written into the memory of the analysis chip 2 together with the data stored in the storage medium in an analyzer (not shown) by using suitable evaluation circuits, for example optoelectronic circuits.
  • the optical gratings 32 were preferably introduced into the microfluidic structure 10 together with the respective points 16 of the biomolecule matrix, which each represent an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.).
  • the biological sample to be examined passes through the microfluidic structure 16 (meandering structure), for example in the direction of the arrows 38.
  • the lens field 36 is located on the side of the grid 32 facing away from the biomolecule matrix. It can, similarly as previously with reference to FIGS. 2 and 3 described, also be provided in the cavity 8 of the analysis chip 2. Likewise, it can consist of a uniform material together with the carrier 4, or, preferably, be provided on the transparent media 12 and 14.
  • analysis chip according to the invention is also suitable for further evaluation methods not described in detail.
  • microfluidic structure 16 as a meandering structure is only exemplary; rather, any other microfluidic structures, such as e.g. a tree structure possible.

Abstract

The invention relates to an analysis and diagnostics instrument - an analysis chip for examining biological samples. Said analysis chip essentially consists of a support on whose surface at least one biomolecular matrix is applied in the form of dots, each dot in the matrix representing an individual species of molecule. The biological sample to be examined is allowed to flow through a microfluidic structure. Preferably at least one medium is provided on the surface of the support, the biomolecular matrix being located on said medium. The analysis chip also has an optical lens field and optionally, optical gratings which are arranged in such a way as to correspond to the biomolecular matrix. An analysis of the biological sample can then be carried out by real-time measurement of the hybridisation with detection of the mass attachment or detection of fluorochrome molecules which were previously built into the probe material.

Description

Analyse- und Diagnostikinstrument Analysis and diagnostic tool
Die Erfindung betrifft ein Analyse- und Diagnostikinstrument zur Untersuchung von biologischen Proben, insbesondere von DNA-Molekülen .The invention relates to an analysis and diagnostic instrument for examining biological samples, in particular DNA molecules.
Im Bereich der biologischen und medizinischen Labortechnik ist eine Reihe von Analyse- bzw. Diagnoεtikinstrumenten bekannt. In bestimmten Ausgestaltungen werden diese üblicherweise auch als Analysechips bezeichnet. Ist der Analysechip zur Untersuchung von DNA-Molekülen vorgesehen, so nennt man ihn DNA-Analysechip, DNA-Array oder DNA-Grid. Die Begriffe Analysechip und Analyse- und Diagnostikinstrument werden im weiteren synonym verwendet .A number of analytical and diagnostic instruments are known in the field of biological and medical laboratory technology. In certain configurations, these are usually also referred to as analysis chips. If the analysis chip is intended for the investigation of DNA molecules, it is called a DNA analysis chip, DNA array or DNA grid. The terms analysis chip and analysis and diagnostic instrument are used synonymously in the following.
Herkömmliche Analysechips bestehen üblicherweise im wesentlichen aus einem Trägermaterial, in dessen Oberfläche Mulden geätzt sind, in die im Fall einer DNA-Analyse verschiedene DNA-Moleküle eingebunden sind. Durch das Inkontaktbringen einer zu untersuchenden DNA-Probe auf die in den Mulden ge- bundenen DNA-Moleküle kann es zu einer Hybridisierung kommen, die Signale über die An- bzw. Abwesenheit und gegebenenfalls die Konzentration von Nucleinsäuren in der Hybridi- sierungslösung liefern. Ein spezieller Analysator liest die Hybridisierungsdaten dann aus. Der Nachweis der Hybridisie- rungsreaktion erfolgt herkömmlicherweise durch optische Verfahren, wie beispielsweise durch Lichteinkopplung über ein Gitter. Bei der Massenanlagerung wird das evaneszente Feld gestört, wodurch eine Störung im Fernfeld meßbar ist. Her- kömmliche Analysechips benutzen als Trägermaterial im allgemeinen Silizium, das ein nicht-transparentes Medium enthält, auf dem die DNA-Moleküle in den Mulden gebunden sind.Conventional analysis chips usually consist essentially of a carrier material, in the surface of which wells are etched, in which, in the case of a DNA analysis, various DNA molecules are incorporated. By bringing a DNA sample to be examined into contact with the bound DNA molecules can lead to hybridization, which provide signals about the presence or absence and possibly the concentration of nucleic acids in the hybridization solution. A special analyzer then reads out the hybridization data. The detection of the hybridization reaction is conventionally carried out by optical methods, such as by coupling light through a grating. The mass accumulation disturbs the evanescent field, whereby a disturbance in the far field can be measured. Conventional analysis chips generally use silicon as the carrier material, which contains a non-transparent medium on which the DNA molecules are bound in the wells.
Analoge, nicht hochintegrierte Systeme, wie ELISA-Systeme, haben insbesondere die Nachteile, daß die zu untersuchende biologische Probe auf jedes Referenzmolekül in den Mulden einzeln aufgebracht werden muß, die Lichteinkopplung zum Nachweis der Hybridisierungsreaktion ineffizient ist, eine Justierung des Analysechips im Analysator schwierig ist, die Verwaltung und Prozessierung von Daten und Parametern aufwendig ist und die Kosten für die Herstellung und den Betrieb sehr hoch sind.Analog, not highly integrated systems, such as ELISA systems, have in particular the disadvantages that the biological sample to be examined has to be applied individually to each reference molecule in the wells, the coupling of light to detect the hybridization reaction is inefficient, and an adjustment of the analysis chip in the analyzer is difficult , the administration and processing of data and parameters is complex and the costs for production and operation are very high.
Der Erfindung liegt daher die Aufgabe zugrunde, ein verbessertes Analyse- bzw. Diagnostikinstrument bzw. -verfahren zur Verfügung zu stellen.The object of the invention is therefore to provide an improved analysis or diagnostic instrument or method.
Zur Lösung dieser Aufgabe geht die Erfindung von dem Grundgedanken aus, einen Analysechip zur Untersuchung von biologischen Proben bereitzustellen, der im wesentlichen aus einem Träger besteht, auf dessen Oberfläche mindestens eine Biomolekülmatrix aus einzelnen Punkten (Dots) auf einen Boden einer mikrofluidischen Struktur, wie z.B. einer Mäanderstruktur, oder eine planare Oberfläche aufgebracht ist. Die zu untersuchende biologische Probe wird an einem Ende der mikrofluidischen Struktur aufgebracht und strömt entlang dieser Struktur zum anderen Ende, wobei die Probe beim Überströmen der jeweiligen Matrixpunkte, die jeweils eine indi- viduelle Molekülspezies repräsentieren, bestimmte Reaktionen auslöst .To achieve this object, the invention is based on the basic idea of providing an analysis chip for examining biological samples, which essentially consists of a carrier, on the surface of which at least one biomolecule matrix composed of individual dots on a bottom of a microfluidic structure, such as, for example a meander structure, or a planar surface is applied. The biological sample to be examined is applied to one end of the microfluidic structure and flows along this structure to the other end, the sample flowing over the respective matrix points, each of which has an individual represent viduelle molecular species, triggers certain reactions.
Diese Reaktion ist, sofern eine Nucleinsäure nachgewiesen werden soll, üblicherweise eine Hybridisierungsreaktion. Je nach Einstellen der Hybridisierungsbedingungen (stringente oder nicht-stringente Hybridisierungsbedingungen) sind Moleküle in der Probe detektierbar, die einen mehr oder weniger großen Ähnlichkeits- bzw. Homologiegrad mit der komplementären Molekülspezies auf der Oberfläche oder einem Teil davon aufweisen. Derartige Hybridisierungsbedingungen hängen beispielsweise von der Temperatur oder der Salzkonzentration in der Probe ab und können vom Fachmann nach üblichen Verfahren eingestellt werden. So kann beispielsweise die Salzkonzentration in einer Probe eingestellt werden. Die Einstellung von Hybridisierungsbedingungen ist im Stand der Technik bekannt und kann z.B. auf der Basis der Lehre in Harnes und Higgins, "Nucleic acid hybridisation, a practical approach" , IRL Press, Oxford 1985 erfolgen.If a nucleic acid is to be detected, this reaction is usually a hybridization reaction. Depending on the setting of the hybridization conditions (stringent or non-stringent hybridization conditions), molecules can be detected in the sample which have a greater or lesser degree of similarity or homology with the complementary molecular species on the surface or a part thereof. Such hybridization conditions depend, for example, on the temperature or the salt concentration in the sample and can be set by the person skilled in the art using customary methods. For example, the salt concentration in a sample can be adjusted. The setting of hybridization conditions is known in the prior art and can e.g. based on the teaching in Harnes and Higgins, "Nucleic acid hybridization, a practical approach", IRL Press, Oxford 1985.
Ist das in der biologischen Probe nachzuweisende Molekül ein Protein oder Peptid, so stehen dem Fachmann eine Reihe von Möglichkeiten zur Verfügung, wie er dieses mit dem erfindungsgemäßen Analysechip nachweisen kann. Eine Möglichkeit ist die Einbindung des Moleküls in einen Antikörper-Sand- wich, wobei der nicht an die Oberfläche fixierte Antikörper (oder ein Derivat davon) vorzugsweise nachweisbar markiert ist .If the molecule to be detected in the biological sample is a protein or peptide, a number of possibilities are available to the person skilled in the art for how he can detect this with the analysis chip according to the invention. One possibility is the incorporation of the molecule into an antibody sandwich, the antibody (or a derivative thereof) that is not fixed to the surface preferably being marked in a detectable manner.
Selbstverständlich sind erfindungsgemäß auch andere Mole- külarten in einer biologischen Probe mit dem Analysechip nachweisbar.Of course, other types of molecules can also be detected in a biological sample using the analysis chip.
Der Begriff "biologische Probe", wie erfindungsgemäß verwendet, bedeutet im weitesten Sinne, daß die Probe biologisches Material wie Nucleinsäuren oder Proteine oder Derivate oder Fragmente davon enthält. Nucleinsäuren können beispielsweise durch Verfahren der rekombinanten DNA-Technologie modifi- ziert, fragmentiert etc. worden sein. Proteine können z.B. aus einer natürlichen Quelle vor der Analyse angereichert worden sein. In einer Ausführungsform wird die Probe aus einer natürlichen Quelle direkt in die Analyse überführt.The term "biological sample" as used according to the invention means in the broadest sense that the sample contains biological material such as nucleic acids or proteins or derivatives or fragments thereof. For example, nucleic acids can be modified by methods of recombinant DNA technology. adorned, fragmented, etc. For example, proteins may have been enriched from a natural source prior to analysis. In one embodiment, the sample is transferred from a natural source directly to the analysis.
Zur erleichterten Datenverwaltung und -Verarbeitung weist der Analysechip ferner ein Speichermedium auf .The analysis chip also has a storage medium for easier data management and processing.
Das erfindungsgemäße Analyse- bzw. Diagnostikinstrument hat gegenüber dem Stand der Technik insbesondere die Vorteile, daß die zu untersuchende Probe nur an einer Stelle auf die mikrofluidische Struktur aufgebracht werden muß, wodurch die Verfahrenszeit erheblich reduziert werden kann, es auf einfache und kostengünstige Weise herstellbar ist, da die mikrofluidische Struktur gleichzeitig mit dem Träger hergestellt werden kann, die theoretische Signalstärke durch die Mikrostrukturierung im Bereich der Proben verstärkt werden kann, ein effizienterer Reaktionsnachweis möglich ist, und/oder fertigungsrelevante und/oder sonstige Daten, wie z.B. Konfiguration, Fertigungslot, Nullkontrolldaten, Patientendaten, Verfallsdatum, Analyseergebnisse und ähnliches direkt auf dem Analysechip gespeichert werden können. Ein weiterer Vorteil besteht darin, daß die Ergebnisse der Analyse, aufgrund einer vorzugsweise mindestens doppelt redun- dant vorhandenen Matrix, mit erheblich höherer Sicherheit festgestellt werden können. Ferner erfolgt die Analyse mit hoher Integrationsdichte und hoher Parallelität. Es sind nur kleine Volumina der Proben erforderlich. Weitere Vorteile sind die Miniaturisierung des Analysechips, die Möglichkeit einer Realtime- und/oder Online-Auswertung zeitgleich mit der Hybridisierungsreaktion.The analysis or diagnostic instrument according to the invention has the advantages over the prior art in particular that the sample to be examined only has to be applied to the microfluidic structure at one point, as a result of which the process time can be considerably reduced and it can be produced in a simple and inexpensive manner , since the microfluidic structure can be produced simultaneously with the carrier, the theoretical signal strength can be increased by the microstructuring in the area of the samples, a more efficient reaction detection is possible, and / or production-related and / or other data, such as Configuration, production lot, zero control data, patient data, expiry date, analysis results and the like can be saved directly on the analysis chip. Another advantage is that the results of the analysis can be determined with considerably greater certainty due to a matrix that is preferably at least double redundant. Furthermore, the analysis is carried out with a high integration density and high parallelism. Only small volumes of the samples are required. Further advantages are the miniaturization of the analysis chip, the possibility of real-time and / or online evaluation at the same time as the hybridization reaction.
Die Erfindung wird im folgenden anhand einer bevorzugten Ausführungsform beispielhaft beschrieben. In den Zeichnungen zeigt: Fig. 1 ein Raumbild eines Analysechips gemäß der Erfindung; Fig. 2 einen Schnitt entlang der Fläche II -II von Fig. 1 zur Verdeutlichung der Lichteinkopplung zum Nachweis von Fluorochrommolekülen; Fig. 3 das Detail III aus Fig. II; undThe invention is described below by way of example with reference to a preferred embodiment. In the drawings: 1 shows a spatial image of an analysis chip according to the invention; FIG. 2 shows a section along the surface II -II of FIG. 1 to illustrate the coupling of light for the detection of fluorochrome molecules; 3 shows the detail III from FIG. II; and
Fig. 4 das Prinzip der Lichteinkopplung zum Nachweis der Massenanlagerung .Fig. 4 shows the principle of light coupling for the detection of mass accumulation.
Das in Fig. 1 dargestellte Analyse- bzw. Diagnostikinstrument 2 - kurz Analysechip - besteht im wesentlichen aus einem Träger oder Tragrahmen 4, der im Querschnitt z.B. ein U-förmiges Profil aufweist, das an seinen Enden geschlossen ist, so daß unter einer Oberfläche 6 ein Hohlraum 8 vorgesehen ist. Auf der Oberfläche 6 des Trägers 4 ist eine mikrofluidische Struktur, z.B. eine Mäanderstruktur 10 vorgesehen, die in den Träger 6 oder bevorzugt in mindestens ein vorzugsweise transparentes Medium 12 und 14 eingelassen ist. Dieses transparente Medium 12 und 14 ist z.B. ein Einsatz aus Glas, Quarzglas, Kunststoff, Silizium, Silikaten. Die Mäanderstruktur 10 ist so ausgebildet, daß darin Punkte (Dots) 16 einer Biomolekülmatrix aufgebracht werden können, wobei jeder Punkt 16 der Matrix vorzugsweise eine individuelle Molekülspezies (DNA, RNA, Proteine, Peptide, Poly- saccharide usw. repräsentiert. Die Mäanderstruktur 10 mit den Dots 16 ist auf dem Analysechip 2 mindestens einmel vorhanden, nämlich auf dem Medium 12, vorzugsweise jedoch mindestens doppelt redundant, also auch mindestens auf dem Medium 14 oder weiteren Medien (nicht dargestellt) .The analysis or diagnostic instrument 2 - in short analysis chip - shown in Fig. 1 consists essentially of a carrier or support frame 4, which in cross section e.g. has a U-shaped profile which is closed at its ends, so that a cavity 8 is provided under a surface 6. On the surface 6 of the carrier 4 there is a microfluidic structure, e.g. a meander structure 10 is provided, which is embedded in the carrier 6 or preferably in at least one preferably transparent medium 12 and 14. This transparent medium 12 and 14 is e.g. an insert made of glass, quartz glass, plastic, silicon, silicates. The meander structure 10 is designed such that dots 16 of a biomolecule matrix can be applied therein, each point 16 of the matrix preferably representing an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.) At least one of the dots 16 is present on the analysis chip 2, namely on the medium 12, but preferably at least twice redundantly, ie also at least on the medium 14 or other media (not shown).
Ebenso ist es möglich, die Oberfläche des Analysechips bzw. des Mediums 12 und 14 planar auszugestalten (ohne mikrofluidische Struktur) und die mindestens eine Biomolekülmatrix darauf vorzusehen. In diesem Fall weist dann eine der Biomolekülmatrix gegenüberliegende Fläche, die das fluidische Volumen mit der zu untersuchenden Probe enthält, die mikrofluidische Struktur auf. Der Analysechip 2 wird dann zur Analyse auf diese Fläche aufgesetzt. Die mikrofluidische Struktur 10 kann ferner sowohl auf der Oberfläche 6 des Analysechips 2 als auch auf der gegenüberliegenden Fläche vorgesehen sein.It is also possible to design the surface of the analysis chip or the medium 12 and 14 to be planar (without a microfluidic structure) and to provide the at least one biomolecule matrix thereon. In this case, a surface opposite the biomolecule matrix, which contains the fluid volume with the sample to be examined, has the microfluidic structure. The analysis chip 2 then becomes Analysis placed on this surface. The microfluidic structure 10 can furthermore be provided both on the surface 6 of the analysis chip 2 and on the opposite surface.
Der Träger 4 des Analysechips ist vorzugsweise ein Kunststoff, der durch Mikrospritzguß hergestellt wird. Als Trägermaterialien eignen sich insbesondere Derivate von Poly- carbonat . Die kovalente Kopplung von Biomolekülen erfolgt vorzugsweise an semifluidischen, gelartigen Polymeroberflächen im Bereich der Proben (z.B. Polymerbrush) . Die Polymeroberflächen besitzen eine eigene Hydrodynamik, wodurch den Polymeroberflächen besondere hydrodynamische Eigenschaften verliehen werden und das Verhältnis zwischen Signal und Hintergrundrauschen positiv beeinflußt wird. Semifluidische Polymeroberflächen sind direkt auf der Oberfläche 6 des Analysechips synthetisiert und können als Nano- bis Mikrostrukturierung von Oberflächen i.a. als Polymerbrushes bezeichnet werden. Die theoretische Signalstärke wird durch die semi- fluidische Oberflächenstrukturierung im Bereich der Proben verstärkt .The carrier 4 of the analysis chip is preferably a plastic, which is produced by micro injection molding. Derivatives of polycarbonate are particularly suitable as carrier materials. The covalent coupling of biomolecules is preferably carried out on semifluidic, gel-like polymer surfaces in the area of the samples (e.g. polymer brush). The polymer surfaces have their own hydrodynamics, which gives the polymer surfaces special hydrodynamic properties and has a positive influence on the relationship between signal and background noise. Semifluidic polymer surfaces are synthesized directly on the surface 6 of the analysis chip and can generally be used as nano- to microstructuring of surfaces. are referred to as polymer brushes. The theoretical signal strength is enhanced by the semi-fluidic surface structuring in the area of the samples.
Zum Betrieb des Analysechips wird z.B. genetisches Material aus einer zu untersuchenden biologischen Probe isoliert, spezifisch amplifiziert und gegebenenfalls markiert. Das so gewonnene Sondenmaterial wird mittels einer Vorrichtung, z.B. einer Pipette oder Spritze, auf ein Ende der Mäanderstruktur 10 des Analysechips 2 aufgebracht. Die zu untersu- chende biologische Probe bzw. das genetische Material überströmt entlang der Mäanderstruktur 10 die einzelnen Punkte 16 der Biomolekülmatrix, die jeweils eine individuelle Mole- külspezies repräsentieren, und hybridisiert im Fall von mindestens teilweise komplementären Biomolekülen. Zur Analyse und Auswertung der Daten des Analysechips 2 stehen dem Fachmann eine Reihe von Verfahren zur Verfügung. Zwei prinzipielle Verfahren werden erfindungsgemäß bevorzugt. Einerseits kann die Auswertung durch eine Echt-Zeit- Messung der Hybridisierung durch Nachweis der Massenanlagerung erfolgen, indem der Analysechip mit einem Gitterkoppler versehen ist, der eine Störung des evaneszenten Felds verursacht. Andererseits besteht die Möglichkeit, durch visuelle oder opto-chemische Mittel nachweisbare Moleküle zwei Fluo- rochrommoleküle nachzuweisen, die z.B. in das Sondenmaterial (DNA, RNA) vor der Analyse eingebaut wurden.To operate the analysis chip, for example, genetic material is isolated from a biological sample to be examined, specifically amplified and, if necessary, marked. The probe material obtained in this way is applied to one end of the meander structure 10 of the analysis chip 2 by means of a device, for example a pipette or syringe. The biological sample to be examined or the genetic material flows over the individual points 16 of the biomolecule matrix along the meandering structure 10, each of which represents an individual molecule species, and hybridizes in the case of at least partially complementary biomolecules. A number of methods are available to the person skilled in the art for analyzing and evaluating the data of the analysis chip 2. Two basic methods are preferred according to the invention. On the one hand, the evaluation can be carried out by a real-time measurement of the hybridization by detecting the mass accumulation by providing the analysis chip with a grating coupler, which causes a disturbance in the evanescent field. On the other hand, there is the possibility of detecting two fluorochrome molecules which can be detected, for example, in the probe material (DNA, RNA) before the analysis by means of molecules which can be detected by visual or opto-chemical means.
Der in den Figuren 1 bis 3 dargestellte Analysechip gemäß der Erfindung ist insbesondere für fluorochrommarkierte Systeme geeignet. Der in Fig. 4 dargestellte Analysechip ist zur Auswertung mit einem Hybridisierungsnachweis durch Massenanlagerung geeignet .The analysis chip according to the invention shown in FIGS. 1 to 3 is particularly suitable for fluorochrome-marked systems. The analysis chip shown in FIG. 4 is suitable for evaluation with a hybridization detection by mass accumulation.
Der Analysechip 2 weist am Außenrand des Trägers 4 Justier- mittel 18, wie z.B. Justiernasen oder -einbuchtungen, auf, so daß der beim Einlegen in einen Analysator (nicht dargestellt) zur Auswertung der Untersuchung genau justierbar ist. Dies ist insbesondere deshalb vorteilhaft, weil die einzelnen Dots 16 der Biomolekülmatrix sowie die Schleifen der Mäanderstruktur 10 vorzugsweise selbst relativ dicht beieinander liegen, z.B. mit einem Abstand zwischen 100 und 500 μm. Es wird sichergestellt, daß die einzelnen Punkte 16 des Analysechips 2 mit entsprechenden Auswerteelementen des Analysators korrespondieren.The analysis chip 2 has adjustment means 18 on the outer edge of the carrier 4, e.g. Adjusting lugs or indentations, so that when inserted into an analyzer (not shown) for evaluating the examination is precisely adjustable. This is particularly advantageous because the individual dots 16 of the biomolecule matrix as well as the loops of the meandering structure 10 are preferably themselves relatively close together, e.g. with a distance between 100 and 500 μm. It is ensured that the individual points 16 of the analysis chip 2 correspond to corresponding evaluation elements of the analyzer.
Der Analysechip 2 weist ferner mindestens ein Speichermedium auf, das Betriebsdaten, fertigungsrelevante Daten wie z.B. Konfiguration, Fertigungslot, Null-Kontrolldaten, eingegebene Patientendaten, das Verfallsdatum, Analyseergebnisse und weitere Hilfsdaten speichern kann. Das Speichermedium ist vorzugsweise ein elektronischer Speicherchip 20, ein Magnetstreifen 22 und/oder ein Barcode 24, kann jedoch auch jedes andere bekannte Speichermedium, wie z.B. ROM oder RAM, sein.The analysis chip 2 also has at least one storage medium which can store operating data, production-relevant data such as configuration, production lot, zero control data, entered patient data, the expiry date, analysis results and other auxiliary data. The storage medium is preferably an electronic one Memory chip 20, a magnetic strip 22 and / or a bar code 24, however, can also be any other known storage medium, such as ROM or RAM.
Der in den Figuren 2 und 3 dargestellte Analysechip 2 weist neben den mit Bezug auf Fig. 1 beschriebenen Merkmalen die für den Nachweis von Fluorochrommolekülen vorteilhaften Merkmale auf .The analysis chip 2 shown in FIGS. 2 and 3 has, in addition to the features described with reference to FIG. 1, the features which are advantageous for the detection of fluorochrome molecules.
Die Analyse einer biologischen Probe in der Mäanderstruktur 10 des Analysechips 2 erfolgt dadurch, daß in einem Analysator der Analysechip 2 von der der Mäanderstruktur 16 abgewandten Seite, d.h. vom Hohlraum 8, beleuchtet wird, was in Form des Pfeils 26 schematisch dargestellt ist. Die Unterseite 28 der Oberfläche 6 ist vorzugsweise mit einem optischen Linsenfeld 30 versehen, das eine konfokale Beleuchtung ermöglicht. Das Linsenfeld 30 ist vorzugsweise so ausgebildet, daß es sich genau unterhalb der Mäanderstruktur 10, insbesondere genau unterhalb der Dots 16 befindet, so daß die in Fig. 3 dargestellte Beleuchtung der einzelnen Dots 16 ermöglicht wird. Das Linsenfeld 30 und der Träger 4 des Analysechips 2 können aus einem einheitlichen Material bestehen, das in einem Schritt z.B. durch Mikrospritzguß hergestellt werden kann. Vorzugsweise befindet sich das Linsenfeld 30 jedoch auf der Unterseite 28 des in den Träger 4 eingelegten transparenten Mediums 12 bzw. 14.The analysis of a biological sample in the meander structure 10 of the analysis chip 2 takes place in that in an analyzer the analysis chip 2 from the side facing away from the meander structure 16, i.e. is illuminated by the cavity 8, which is shown schematically in the form of arrow 26. The underside 28 of the surface 6 is preferably provided with an optical lens field 30, which enables confocal illumination. The lens array 30 is preferably designed such that it is located exactly below the meandering structure 10, in particular exactly below the dots 16, so that the illumination of the individual dots 16 shown in FIG. 3 is made possible. The lens field 30 and the carrier 4 of the analysis chip 2 can consist of a uniform material, which in one step e.g. can be produced by micro injection molding. However, the lens field 30 is preferably located on the underside 28 of the transparent medium 12 or 14 inserted into the carrier 4.
Die Beleuchtung 26 von der Unterseite 28 des Analysechips 2 bringt insbesondere den Vorteil, daß die Intensität der Lichtquelle reduziert werden kann, da die Lichtstrahlen keinen Strahlteiler durchlaufen müssen. Das einfallende Licht 26 wird durch die Linsen des Linsenfelds 30 auf die einzelnen Punkte 16 der Biomolekülmatrix fokussiert, wodurch die Dots 16, die mit der zu untersuchenden Probe eine Reaktion hervorgerufen haben, durch die Fluorochrommoleküle Fluoreszenz bewirken. Weitere Möglichkeiten zum Nachweis der Moleküle beruhen auf dem Prinzip der Biolumineszenz, z.B. Phosphoreszenz, radioaktiven Markierungen, die durch Auflegen von Röntgenfilmen nachgewiesen werden können. Durch eine geeignete Auswerteschaltung, z.B. eine opto- elektronische Schaltung, kann dann eine weitere Verarbeitung der gewonnenen Daten bezüglich der zu analysierende Probe erfolgen. Diese Daten können zusammen mit dem in dem Speichermedium gespeicherten Daten weiter verarbeitet und/oder in den Speicher des Analysechips 2 geschrieben werden.The illumination 26 from the underside 28 of the analysis chip 2 has the particular advantage that the intensity of the light source can be reduced since the light beams do not have to pass through a beam splitter. The incident light 26 is focused by the lenses of the lens array 30 onto the individual points 16 of the biomolecule matrix, as a result of which the dots 16, which have caused a reaction with the sample to be examined, cause fluorescence through the fluorochrome molecules. Other ways to prove the Molecules are based on the principle of bioluminescence, eg phosphorescence, radioactive labels, which can be detected by applying X-ray films. A suitable evaluation circuit, for example an optoelectronic circuit, can then be used to further process the data obtained with respect to the sample to be analyzed. This data can be further processed together with the data stored in the storage medium and / or written into the memory of the analysis chip 2.
Fig. 4 zeigt die Lichteinkopplung für Analysechips, bei denen der Hybridisierungsnachweis über den Nachweis der Massenanlagerung erfolgt. Dazu wird ein optisches Verfahren benützt, bei dem die Lichteinkopplung über eine Vielzahl von Gittern 32 erfolgt, wodurch das evaneszente Feld beeinflußt wird. Diese Beeinflussung ist im Fernfeld 39 meßbar. Jedem Punkt 16 der Biomolekülmatrix in der mikrofluidischen Struktur 10 ist exakt ein optisches Gitter 32 zugeordnet, so daß die Effizienz der Lichteinkopplung erheblich erhöht werden kann. Das Licht 34 einer Beleuchtungsquelle wird über ein zweidimensionales Linsenfeld 36 oder über ein zweidimensio- nales Fresnel-Linsenfeld, z.B. ein Hologramm oder diffrak- tive Optik, auf die in der Matrix angeordneten Gitter 32 fo- kussiert. Dadurch kann ein effizienter Hybridisierungsnachweis durch den Nachweis der Massenanlagerung erzielt werden, wenn eine Auswertung des Fernfelds erfolgt . Die gewonnenen Analysedaten können zusammen mit den im Speichermedium gespeicherten Daten in einem Analysator (nicht dargestellt) durch Verwendung geeigneter Auswerteschaltungen, z.B. optoelektronische Schaltungen, weiter, verarbeitet und/oder in den Speicher des Analysechips 2 geschrieben werden. Die optischen Gitter 32 wurden vorzugsweise zusammen mit den jeweiligen Punkten 16 der Biomolekülmatrix, die jeweils eine individuelle Molekülspezies (DNA, RNA, Proteine, Peptide, Polysaccharide usw.) repräsentieren, in die mikrofluidische Struktur 10 eingebracht. Die zu untersuchende biologische Probe durchläuft die mikrofluidische Struktur 16 (Mäanderstruktur) z.B. in Richtung der Pfeile 38. das Linsenfeld 36 befindet sich auf der von der Biomolekülmatrix abgewandten Seite des Gitters 32. Es kann, ähnlich wie zuvor mit Bezug auf die Figuren 2 und 3 beschrieben, ebenfalls im Hohlraum 8 des Analysechips 2 vorgesehen sein. Ebenso kann es zusammen mit dem Träger 4 aus einem einheitlichen Material bestehen, oder, vorzugsweise, an den transparenten Medien 12 und 14 vorgesehen sein.FIG. 4 shows the coupling of light for analysis chips in which the hybridization detection takes place via the detection of the mass accumulation. For this purpose, an optical method is used in which the light is coupled in via a large number of gratings 32, as a result of which the evanescent field is influenced. This influence can be measured in the far field 39. Exactly one optical grating 32 is assigned to each point 16 of the biomolecule matrix in the microfluidic structure 10, so that the efficiency of the light coupling can be increased considerably. The light 34 from an illumination source is focused on the grating 32 arranged in the matrix via a two-dimensional lens field 36 or via a two-dimensional Fresnel lens field, for example a hologram or diffractive optics. As a result, an efficient hybridization detection can be achieved by the detection of the mass accumulation when the far field is evaluated. The analysis data obtained can be further processed and / or written into the memory of the analysis chip 2 together with the data stored in the storage medium in an analyzer (not shown) by using suitable evaluation circuits, for example optoelectronic circuits. The optical gratings 32 were preferably introduced into the microfluidic structure 10 together with the respective points 16 of the biomolecule matrix, which each represent an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.). The biological sample to be examined passes through the microfluidic structure 16 (meandering structure), for example in the direction of the arrows 38. The lens field 36 is located on the side of the grid 32 facing away from the biomolecule matrix. It can, similarly as previously with reference to FIGS. 2 and 3 described, also be provided in the cavity 8 of the analysis chip 2. Likewise, it can consist of a uniform material together with the carrier 4, or, preferably, be provided on the transparent media 12 and 14.
Neben den mit Bezug auf die Figuren 2 bis 4 beschriebenen Auswerteverfahren zur Auswertung der Analysechips 2 ist der erfindungsgemäße Analysechip ebenso für weitere, nicht näher beschriebene Auswerteverfahren geeignet.In addition to the evaluation methods for evaluating the analysis chips 2 described with reference to FIGS. 2 to 4, the analysis chip according to the invention is also suitable for further evaluation methods not described in detail.
Die Ausbildung der mikrofluidischen Struktur 16 als Mäanderstruktur ist lediglich exemplarisch, es sind vielmehr auch beliebige andere mikrofluidische Strukturen, wie z.B. eine Baumstruktur möglich. The formation of the microfluidic structure 16 as a meandering structure is only exemplary; rather, any other microfluidic structures, such as e.g. a tree structure possible.

Claims

Patentansprüche claims
Analysechip (2) zur Untersuchung von biologischen Pro¬ ben, der einen Träger (4) aufweist, auf dessen Oberflä¬ che (6) mindestens eine Biomolekülmatrix (16) durch ko- valente Kopplung von Biomolekülen an semifluidische, gelartige Polymeroberflächen im Bereich der Proben auf¬ gebracht ist . A nalysechip (2) for the investigation of biological Pro ¬ ben having a support (4), on whose Oberflä ¬ surface (6) at least one Biomolekülmatrix (16) by covalent coupling of biomolecules to semifluidische, gel-like polymer surfaces in the area of Samples is brought on ¬ .
Analysechip (2) zur Untersuchung von biologischen Proben, der einen Träger (4) aufweist, auf dessen Oberfläche (6) mindestens eine Biomolekülmatrix (16) aufgebracht ist, wobei die zu untersuchende biologische Probe über jede Biomolekülmatrix (16) strömt und der Analysechip (2) mindestens ein Speichermedium (20, 22, 24) auf eist .Analysis chip (2) for examining biological samples, which has a carrier (4) on whose surface (6) at least one biomolecule matrix (16) is applied, the biological sample to be examined flowing over each biomolecule matrix (16) and the analysis chip ( 2) at least one storage medium (20, 22, 24).
3. Analysechip (2) nach Anspruch 1 oder 2, wobei der Träger (4) aus einem Kunststoff hergestellt ist.3. Analysis chip (2) according to claim 1 or 2, wherein the carrier (4) is made of a plastic.
4. Analysecip (2) nach Anspruch 3, wobei der Kunststoff insbesondere ein Derivat von Polycarbonat ist.4. Analysis clip (2) according to claim 3, wherein the plastic is in particular a derivative of polycarbonate.
5. Analysecip (2) nach Anspruch 4, wobei die Biomolekülmatrix (16) durch kovalente Kopplung von Biomolkülen an semifluidische, gelartige Polymeroberflächen im Bereich der Proben aufgebracht ist.5. Analysis clip (2) according to claim 4, wherein the biomolecule matrix (16) is applied by covalent coupling of biomolecules to semifluidic, gel-like polymer surfaces in the region of the samples.
6. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei die Biomolekülmatrix einzelne Dots (16) aufweist.6. Analysis chip (2) according to one of the preceding claims, wherein the biomolecule matrix has individual dots (16).
7. Analysechip (2) nach Anspruch 6, wobei jeder Dot (16) der Biomolekülmatrix eine individuelle Molekülspezies repräsentiert . 7. Analysis chip (2) according to claim 6, wherein each dot (16) of the biomolecule matrix represents an individual molecular species.
8. Analysechip (2) nach einem der Ansprüche 1 bis 7, wobei die Oberfläche (6) , auf der die Biomolekülmatrix (16 aufgebracht ist, planar ist.8. Analysis chip (2) according to one of claims 1 to 7, wherein the surface (6) on which the biomolecule matrix (16) is applied is planar.
9. Analysechip (2) nach einem der Ansprüche 1 bis 7, wobei die Oberfläche (6) eine mikrofluidische Struktur (10) aufweist, in der die Biomoleküle vorgesehen sind.9. Analysis chip (2) according to one of claims 1 to 7, wherein the surface (6) has a microfluidic structure (10) in which the biomolecules are provided.
10. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei der Träger (4) mindestens ein Medium (12, 14) aufweist, auf dem sich die Biomolekülmatrix (16) befindet.10. Analysis chip (2) according to one of the preceding claims, wherein the carrier (4) has at least one medium (12, 14) on which the biomolecule matrix (16) is located.
11. Analysechip (2) nach Anspruch 10, wobei das Medium (12, 14) Glas, Quarzglas, ein Kunststoff oder Silicium-Sili- kat ist.11. Analysis chip (2) according to claim 10, wherein the medium (12, 14) is glass, quartz glass, a plastic or silicon silicate.
12. Analysechip (2) nach einem der Ansprüche 1 bis 11, wobei der Analysechip ferner mindestens ein Speichermedium12. Analysis chip (2) according to one of claims 1 to 11, wherein the analysis chip further at least one storage medium
(20, 22, 24) aufweist.(20, 22, 24).
13. Analysechip (2) nach einem der Ansprüche 1 bis 12 oder 12, wobei das Speichermedium ein elektronischer Speicherchip (20), ein Magnetstreifen (22) und/oder ein Barcode (24) ist.13. Analysis chip (2) according to one of claims 1 to 12 or 12, wherein the storage medium is an electronic memory chip (20), a magnetic strip (22) and / or a bar code (24).
14. Analysechip (2) nach einem der vorherigen Ansprüche, wobei der Träger (4) ferner Justiermittel (18) zum exakten Einlegen in einen Analysator aufweist.14. Analysis chip (2) according to one of the preceding claims, wherein the carrier (4) further comprises adjustment means (18) for exact insertion into an analyzer.
15. Analysechip (2) nach einem der Ansprüche 3 bis 14, wobei der Träger (4) ein Mikrospritzguß-Element ist.15. Analysis chip (2) according to one of claims 3 to 14, wherein the carrier (4) is a micro-injection molding element.
16. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei die Biomolekülmatrix (16) mindestens doppelt re- dundant vorhanden ist. 16. Analysis chip (2) according to one of the preceding claims, wherein the biomolecule matrix (16) is at least twice redundant.
17. Analysechip (2) nach einem der Ansprüche 9 bis 16, wobei die mikrofluidische Struktur (10) einen mäanderförmigen Kanal bildet.17. Analysis chip (2) according to one of claims 9 to 16, wherein the microfluidic structure (10) forms a meandering channel.
18. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei an einer Unterseite (28) des Trägers (4) ein optisches Linsenfeld (30; 36) vorgesehen ist.18. Analysis chip (2) according to one of the preceding claims, wherein an optical lens field (30; 36) is provided on an underside (28) of the carrier (4).
19. Analysechip (2) nach Anspruch 18, wobei das Linsenfeld (36) ein zweidimensionales Fresnel-Linsenfeld ist.19. Analysis chip (2) according to claim 18, wherein the lens field (36) is a two-dimensional Fresnel lens field.
20. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei entsprechend der Biomolekülmatrix (16) eine Vielzahl optischer Gitter (32) vorgesehen ist.20. Analysis chip (2) according to one of the preceding claims, wherein a plurality of optical gratings (32) is provided corresponding to the biomolecule matrix (16).
21. Analysechip (2) nach einem der vorhergehenden Ansprüche, wobei zur Auswertung der biologischen Proben der Analysechip (2) so ausgestaltet ist, daß die Biomolekülmatrix (16) von einer der der Oberfläche (6) gegenüberliegenden Unterseite (28) mit Lichtstrahlen (26; 34) von einer Lichtquelle beaufschlagt werden kann.21. Analysis chip (2) according to one of the preceding claims, wherein for the evaluation of the biological samples, the analysis chip (2) is designed such that the biomolecule matrix (16) from one of the underside (28) opposite the surface (6) with light rays (26 ; 34) can be acted upon by a light source.
22. Verwendung des Analysechips (2), insbesondere nach einem der vorhergehenden Ansprüche, zur Untersuchung von DNA-, RNA-Molekülen, Proteinen, Peptiden und/oder Polysaccha- riden.22. Use of the analysis chip (2), in particular according to one of the preceding claims, for the investigation of DNA, RNA molecules, proteins, peptides and / or polysaccharides.
23. Verfahren zur Analyse von Biomolekülen auf einem Analysechip, insbesondere nach einem der vorhergehenden Ansprüche, mit den folgenden Verfahrensschritten: (a) Aufbringen einer zu analysierenden Probe auf eine23. A method for analyzing biomolecules on an analysis chip, in particular according to one of the preceding claims, with the following method steps: ( a ) applying a sample to be analyzed to a
Oberfläche (6) , die mindestens eine Biomolekülmatrix (16) aufweist; (b) Überströmen der Biomolekülmatrix (16) durch die Probe entlang einer mikrofluidischen Struktur; (c) Einlegen des Analysechips (2) in einen Analysator; und (d) Auswerten des Analysechips (2) . Surface (6) which has at least one biomolecule matrix (16); (b) overflow of the biomolecule matrix (16) through the sample along a microfluidic structure; ( c ) inserting the analysis chip (2) into an analyzer; and (d) evaluating the analysis chip (2).
24. Verfahren nach Anspruch 23, wobei die Auswertung des Analysechips (2) durch Einstrahlen von Licht von einer Unterseite (28) des Analysechips (2) erfolgt.24. The method according to claim 23, wherein the analysis of the analysis chip (2) is carried out by irradiation of light from an underside (28) of the analysis chip (2).
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25. Verfahren nach Anspruch 23 oder 24, wobei die Auswertung des Analysechips (2) durch Lichteinkopplung über ein optisches Linsenfeld (36) und Gitter (32) erfolgt, wobei jedes Gitter (32) jeweils einem Punkt (16) der Biomole- ° külmatrix zugeordnet ist und das optische Linsenfeld (36) die einfallenden Lichtstrahlen auf die jeweiligen Gitter (32) fokussiert, so daß bei einer Massenanlagerung das evaneszente Feld beeinflußt und diese Beeinflussung im Fernfeld meßbar wird. 525. The method according to claim 23 or 24, wherein the analysis of the analysis chip (2) is carried out by coupling light through an optical lens field (36) and grating (32), each grating (32) each having a point (16) of the biomolecule matrix is assigned and the optical lens field (36) focuses the incident light rays onto the respective grating (32), so that the mass is influenced by the evanescent field and this influence can be measured in the far field. 5
26. Verfahren nach Anspruch 23 oder 24, wobei die Auswertung des Analysechips (2) durch Lichteinkopplung über ein Linsenfeld (36) erfolgt, das am Analysechip (2) integriert ist und eine konfokale Beleuchtung bewirkt, so 0 daß entsprechend markierte Systeme einen optisch erkennbaren Reaktionsnachweis liefern.26. The method according to claim 23 or 24, wherein the analysis of the analysis chip (2) by light coupling via a lens field (36) which is integrated on the analysis chip (2) and causes confocal illumination, so that 0 marked systems are optically recognizable Provide proof of reaction.
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PCT/EP1999/002918 1998-04-30 1999-04-29 Instrument for analysis and diagnostics WO1999057310A2 (en)

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AU38252/99A AU3825299A (en) 1998-04-30 1999-04-29 Instrument for analysis and diagnostics

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DE19819537.0 1998-04-30
DE1998119537 DE19819537A1 (en) 1998-04-30 1998-04-30 Analysis and diagnostic tool

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WO1999057310A2 true WO1999057310A2 (en) 1999-11-11
WO1999057310A3 WO1999057310A3 (en) 2000-03-09

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WO2003008974A1 (en) * 2001-07-18 2003-01-30 Micronas Gmbh Biosensor and method for detecting analytes by means of time-resolved luminescence
WO2003080789A1 (en) * 2002-03-27 2003-10-02 Micronas Gmbh Device and method for detecting cellular processes by means of luminescence measurements
WO2003082730A1 (en) * 2002-03-31 2003-10-09 Gyros Ab Efficient mmicrofluidic devices
WO2003104384A1 (en) * 2002-06-05 2003-12-18 Bioprocessors Corporation Reactor systems having a light-interacting component
WO2004052540A3 (en) * 2002-12-05 2004-09-16 Protasis Corp Configurable microfluidic substrate assembly
WO2007050040A1 (en) * 2005-10-28 2007-05-03 Agency For Science, Technology And Research Immobilization unit and device for isolation of nucleic acid molecules

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DE10312670A1 (en) * 2003-03-21 2004-10-07 Friz Biochem Gmbh Substrate for controlled wetting of predetermined wetting points with small liquid volumes, substrate cover and flow chamber
US8221843B2 (en) 2005-02-15 2012-07-17 Lisa Dräxlmaier GmbH Methods and compositions for coating interior components of motor vehicles and interior components of motor vehicles coated using same
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US5284622A (en) * 1991-10-02 1994-02-08 Boehringer Mannheim Gmbh Test carrier for the analysis of fluids
US5770721A (en) * 1993-08-11 1998-06-23 University Of Chicago Method of manufacturing a matrix for the detection of mismatches
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003008974A1 (en) * 2001-07-18 2003-01-30 Micronas Gmbh Biosensor and method for detecting analytes by means of time-resolved luminescence
WO2003080789A1 (en) * 2002-03-27 2003-10-02 Micronas Gmbh Device and method for detecting cellular processes by means of luminescence measurements
US7767443B2 (en) 2002-03-27 2010-08-03 Micronas Gmbh Device and method for detecting cellular processes by means of luminescence measurements
WO2003082730A1 (en) * 2002-03-31 2003-10-09 Gyros Ab Efficient mmicrofluidic devices
WO2003104384A1 (en) * 2002-06-05 2003-12-18 Bioprocessors Corporation Reactor systems having a light-interacting component
WO2004052540A3 (en) * 2002-12-05 2004-09-16 Protasis Corp Configurable microfluidic substrate assembly
WO2007050040A1 (en) * 2005-10-28 2007-05-03 Agency For Science, Technology And Research Immobilization unit and device for isolation of nucleic acid molecules

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WO1999057310A3 (en) 2000-03-09
AU3825299A (en) 1999-11-23
DE19819537A1 (en) 2000-03-16

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