EP2577311A1 - Procédé et dispositif d'examen optique - Google Patents

Procédé et dispositif d'examen optique

Info

Publication number
EP2577311A1
EP2577311A1 EP11721538.4A EP11721538A EP2577311A1 EP 2577311 A1 EP2577311 A1 EP 2577311A1 EP 11721538 A EP11721538 A EP 11721538A EP 2577311 A1 EP2577311 A1 EP 2577311A1
Authority
EP
European Patent Office
Prior art keywords
light
liquid
optically active
detection
surface area
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP11721538.4A
Other languages
German (de)
English (en)
Inventor
Gert Blankenstein
Dirk Kurowski
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Boehringer Ingelheim Microparts GmbH
Original Assignee
Boehringer Ingelheim Microparts GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Boehringer Ingelheim Microparts GmbH filed Critical Boehringer Ingelheim Microparts GmbH
Priority to EP11721538.4A priority Critical patent/EP2577311A1/fr
Publication of EP2577311A1 publication Critical patent/EP2577311A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • 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
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54346—Nanoparticles
    • 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
    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/588—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with semiconductor nanocrystal label, e.g. quantum dots
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0654—Lenses; Optical fibres
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/16—Surface properties and coatings
    • B01L2300/168—Specific optical properties, e.g. reflective coatings

Definitions

  • the present invention relates to a method for optical examination according to the preamble of claim 1 and an apparatus according to the preamble of claim 14.
  • the present invention is concerned with the optical examination of a surface area, preferably for the optical detection of a reaction proceeding thereon or of a substance bound thereto, in particular with the diagnostics in the case of microfluidic samples.
  • the present invention is concerned with preferably miniaturized immunoassays. So the study of samples using antibodies.
  • the present invention particularly preferably relates to so-called cartridge concepts, that is to say small, in particular card-type, devices for examining a preferably liquid sample or realizing immunoassays.
  • an analyte to be determined of a sample is bound to a surface region, in particular by means of an antibody.
  • the binding of the analyte or other substance by means of an antibody is also referred to in the present invention as an immunoassay reaction.
  • the analyte is combined with a detectable, in particular fluorescent conjugate or other detection partner before or after binding to the surface region.
  • a complex is formed from the analyte and the conjugate, which binds to the immobilized antibody on the Oberfi Stahl.
  • the analyte to be investigated or to be determined binds to the antibody and binds the conjugate to antibodies not occupied by an analyte.
  • the detection of the conjugate or other detection partner for determining the analyte is carried out optically, in particular by measuring the luminescence or fluorescence.
  • This is known, for example from WO 02/08762 AI.
  • a surface region to be examined is irradiated with laser light, and emitted light, namely emitted fluorescent light, is detected by means of a CCD camera.
  • a bandpass filter is used to block the laser light while transmitting the light emitted by fluorescent molecules. This significantly improves the signal-to-noise ratio and is especially important for small signals, ie low emission intensities.
  • such a band filter is arranged directly in front of the camera or another receiver for the emitted or emitted light. In this way, however, unwanted noise or background light or the like can not be turned off.
  • WO 02/14926 A2 deals with fluidic systems in which light rays are manipulated by fluids which reflect, bend, optically filter or scatter the light, for example to realize optical switches or filters. However, WO 02/14926 A2 does not deal with the optical examination of surfaces or the detection of substances on surfaces.
  • the surface area to be examined is covered by an optically active liquid which filters, reflects, scatters, polarizes and / or absorbs light having a wavelength at least substantially corresponding to the light which is input and / or radiated.
  • the surface region can be examined optically, in particular from the side facing away from the liquid, and the desired optical detection or determination can be carried out, particularly preferably in the form of a luminescence or fluorescence. cenzunk, wherein the optically active liquid unwanted spurious signals, such as reflections on an opposite chamber wall, emissions of substances on other walls, emissions of the wall material. Background influences or the like, surprisingly effective suppressed or can hide.
  • an extraordinarily improved signal-to-noise ratio can be achieved.
  • a significantly improved linearity can be achieved in the detection or determination of an analyte or in the realization of an immunoassay.
  • the irradiation of the light is preferably carried out by the wall forming or supporting the surface area, that is to say in particular from the side facing away from the liquid.
  • the emitted light is preferably also detected through this wall, ie in particular on the side facing away from the liquid. Accordingly, a radiation of the liquid is not required. As a result, further unwanted suggestions or emissions or other disorders in deeper areas - for example, on a chamber floor - avoided or at least minimized.
  • the optical examination or detection is carried out by a luminescence or fluorescence measurement.
  • a luminescence or fluorescence measurement particularly preferably UV light is irradiated. This allows a high degree of independence from any interfering light sources.
  • the optically active liquid preferably contains at least one dye and / or pigments or particles.
  • a very effective absorption or scattering or filtering in particular in the visible range in which the emitted or emitted light is usually or preferably located, and / or in the UV range in which the incident light is usually or preferably, achieved become.
  • the optically active liquid is particularly preferably a washing liquid or other reaction liquid which is used in particular for the formation and / or binding of a substance and / or for an immunoassay reaction.
  • a washing liquid or other reaction liquid which is used in particular for the formation and / or binding of a substance and / or for an immunoassay reaction.
  • the liquid preferably covers the surface area or detection area to be examined with a layer thickness of more than 0.1 ⁇ m, preferably more than 1 ⁇ m, in particular more than 10 ⁇ m, preferably more than 50 microns, more preferably more than 100 microns.
  • the device according to the invention is designed in particular for the realization of the aforementioned method and / or as an immunoassay. For this purpose, it has a particularly micro-fluidic detection chamber, which in turn is provided with at least one transparent wall with a chamber-side surface area.
  • the surface area or a substance arranged thereon or bonded, such as a detection partner or conjugate, can be irradiated with irradiated light and / or light emitted therefrom can be detected.
  • the covering of the surface area by the optically active liquid is preferably carried out by filling the detection chamber with the liquid. This allows a very simple implementation.
  • microfluidic are inventions according to volumes of preferably less than 10 ml, more preferably less than 1 ml, and / or chamber or liquid cross-sections (maximum or hydraulic diameter) of preferably less than 2 mm, more preferably less than 500 microns , to understand.
  • the surface area to be investigated serves in particular at least for the detection or binding of a substance.
  • This substance may be an analyte of a sample or a complex formed therefrom or its dependent reaction product and / or a reagent that interacts or binds with the sample, an analyte of the sample, a complex thereof or the like.
  • the reagent may interact or bind with a complex of the analyte or with an analyte-dependent reaction product.
  • the reagent itself is fixed or immobilized in or on the surface area.
  • the immobilized with an Antibody which interacts with an analyte of the sample or a complex containing the analyte, particularly preferably binds.
  • the reagent is formed by the antibody, which interacts directly or indirectly with the analyte, in particular with a complex containing the analyte or the like.
  • the term "interacting" is preferably to be understood in a broad sense in the present invention,
  • the surface region is at least substantially planar or smooth and / or arranged on an at least substantially planar fluid side or chamber inner side of a chamber wall. This is a good or defined detection or determination of an analyte and a simple structure and reaction process beneficial.
  • the term "determination" in the present invention is preferably understood as meaning the detection of an interaction, modification or reaction on the surface region and / or binding of a substance, such as an analyte, complex or the like, on the surface region in order to obtain a preferably qualitative and / or quantitative examination of the sample, in particular a qualitative and / or quantitative determination of at least one analyte of the sample to allow.
  • a substance such as an analyte, complex or the like
  • an optical detection or measurement in particular of a preferably fluorescent detection partner, conjugate or the like, to allow the determination.
  • the optical detection is particularly preferably carried out by a luminescence or fluorescence measurement.
  • Fig. 1 is a schematic plan view of a proposed device with a detection chamber
  • FIG. 2 shows a schematic cross-section of the detection chamber during an examination
  • FIG. and Fig. 3 is a schematic cross section of the detection chamber during another examination.
  • the device 1 shows a schematic plan view of a proposed device 1.
  • the device 1 is preferably at least substantially card-like, plate-like, flat, thin and / or flat.
  • the device according to the preamble 1 is preferably designed as an immunoassay or designed to carry out an immunoassay reaction.
  • the device 1 preferably has a sample receptacle 2 for a liquid sample 3. It is used in particular to examine the sample 3.
  • the sample 3 is preferably-in particular at least partially or substantially-liquid, wherein individual analytes or substances to be determined in the sample 3, such as proteins or the like, themselves are not liquid or must be.
  • the sample 3 may be, for example, a body fluid to be examined, saliva, blood or the like.
  • the device 1 or sample receptacle 2 may, for example, contain a filter in order to separate off blood plasma or the like and to examine it in the device 1.
  • a filter in order to separate off blood plasma or the like and to examine it in the device 1.
  • the device 1 preferably has a channel 4, which forwards the sample 3 or a component thereof, such as blood plasma, in particular to a mixing chamber 5 of the device 1.
  • the sample 3 is preferably mixed or mixed with a detection partner, in particular a conjugate.
  • the detection partner or the conjugate can in particular connect to an analyte to be determined, in particular to form a complex. This step is also called incubation.
  • the detection partner or the conjugate can be present, for example, in a dried form in the mixing chamber 5 or in another area of the device 1 and be detached from the supplied sample 3.
  • the detection partner or the conjugate may also be present or presented as required in liquid form or supplied.
  • a receptacle 6 for a detection liquid 7 is schematically indicated, which is connected for example via a channel 8 to the mixing chamber 5.
  • the sample 3 and the detection liquid 7 can be combined in the mixing chamber 5 and mixed or mixed to achieve the desired incubation.
  • the preferably already incubated sample 3 is supplied by means of a channel 9 of a detection chamber 10 of the device I.
  • a surface-bound reaction or immunoassay reaction takes place in the detection chamber 10.
  • the detection chamber 10 is used in particular for an optical examination or detection of the reaction or the sample 3 or a surface area, particularly preferably the optical detection or measurement of an analyte of the sample 3 or another substance or another, in particular related reactions or the like will be discussed later.
  • a binding of a substance, such as the analyte or the like to be determined takes place at the surface area, in particular It prefers by means of or to antibodies.
  • the antibodies are immobilized on the surface area in the detection chamber 10.
  • other constructive or reactive implementations are possible.
  • the device 1 is preferably designed such that after the aforementioned binding or at the conclusion of the immunoassay reaction or another reaction in the detection chamber 10, a washing step or generally rinsing can take place.
  • the device 1 preferably has a receptacle or a reservoir 1 1 for a washing liquid 12 or other rinsing liquid.
  • the reservoir 1 1 is indirectly or directly connected to the detection chamber 10 via a channel 13, for example, via the channel 9 in the illustration.
  • the device 1 may have corresponding valves.
  • a valve 14 (in particular in channel 9) for controlling the supply of the sample 3 and optionally reaction liquid into the detection chamber 10 and / or a valve 15 (in particular in channel 13). be provided for controlling the supply of the washing liquid 12 in the detection chamber 10.
  • the channel 13 preferably opens into the channel 9 downstream of the valve 14.
  • valves can also be provided in the channels 4 and 8.
  • a preferred sequence provides that first the sample 3 to be examined is incubated with the detection partner or conjugate in the mixing chamber 5 for a predetermined time and then transferred, for example by opening the valve 14, into the detection chamber 10. There, for example, the analyte, a complex formed from analyte and conjugate or another substance can bind. After a particularly predetermined time, the washing or rinsing then takes place, in particular by opening the valve 15. The excess fluid volumes can be forwarded from the detection chamber 10 into a connected excess reservoir 16, often also called waste, to the device 1. However, other constructive solutions are possible.
  • the device 1 or the detection chamber 10 is in particular a microfluidic system.
  • individual liquids or all liquids flow through the device 1 at least in regions due to capillary forces, particularly preferably exclusively by capillary forces.
  • other forces for example due to pressure differences, may also be used, in particular in order to enable or ensure a desired process or reaction sequence.
  • individual valves by exerting a corresponding pressure, for example, on the receptacle 6 or the reservoir 1 1, are opened.
  • the device 1 or detection chamber 10 is preferably constructed from a base part or lower part 17, which in the illustrated example is particularly preferably designed as a spritz casting part and / or channel plate with corresponding recesses, recesses or the like.
  • a base part or lower part 17 which in the illustrated example is particularly preferably designed as a spritz casting part and / or channel plate with corresponding recesses, recesses or the like.
  • wells for the Probenaufiiahme 2 the channel 4, the mixing chamber 5, the receptacle 6, the channel 8, the channel 9, the detection chamber 10, a connection channel from the detection chamber 10 to the overflow reservoir 16 and / or the overflow reservoir 16 itself are formed ,
  • the base part or lower part 17 is preferably covered by an upper part or a cover, in particular at least substantially over the entire surface and / or throughout, for example, an opening or opening in the region of the sample holder 2 for receiving the sample 3 and / or for example a ventilation or Vent (vent) in particular in conjunction with the overflow reservoir 16 may be formed.
  • an opening or opening in the region of the sample holder 2 for receiving the sample 3 and / or for example a ventilation or Vent (vent) in particular in conjunction with the overflow reservoir 16 may be formed.
  • the receptacle 6 and the reservoir 1 1 may also be covered or closed by the upper part or cover element or other cover , in particular after filling the detection liquid 7 or washing liquid 12.
  • the upper part or the lid can therefore also be constructed in several parts.
  • the upper part or cover element 18 may, if necessary, be formed by film or any other suitable material.
  • the connection to the base part or lower part 17 is preferably carried out by gluing, sealing, welding, laminating, pressing, clamping, riveting and / or in any other suitable manner.
  • the device 1 is designed in particular for optical examination or detection.
  • a corresponding, proposed method for the optical examination of a surface area or for the optical detection or determination of an analyte or other substance or an immunoassay reaction are explained in more detail below with reference to FIGS. 2 and 3.
  • FIGS. 2 and 3 show schematic, fragmentary cross sections of the detection chamber 10 along the line S of FIG. 1.
  • FIG. 2 schematically shows the detection chamber 10 during a first examination of a surface area 19 of the detection chamber 10 or a wall of the detection chamber 10.
  • the wall or the surface area 19 is preferably formed by the cover element 18 in the illustrated embodiment.
  • the wall or the cover element 18 is preferably transparent (sufficient for the optical examination or detection).
  • the surface area 19 to be examined lies on the fluid side of the detection chamber 10.
  • the wall or the surface area 19 represents in particular an immediate boundary of the inner space of the detection chamber 10 or for liquid.
  • the detection chamber 10 is filled with the washing liquid 12 for or during the optical examination.
  • the detection chamber 10 can in principle also be filled with any other liquid for the optical examination. Accord- In the following, only the term "liquid 12" will be used.
  • an immunoassay reaction has preferably already taken place in the deletion chamber 30 or a substance to be detected is already bound to the surface region 19.
  • the surface region 19 is (at least partially) provided with immobilized binding partners, in this case antibodies 20.
  • immobilized binding partners in this case antibodies 20.
  • This is a substance to be detected or an analyte 21 with an associated detection partner or conjugate 22 or a complex formed therefrom or the like., Bound.
  • This so-called “sandwich structure” is shown very schematically and enlarged in FIG.
  • the immunoassay reaction proceeds in the illustrated example in particular as follows:
  • the sample 3 with the analyte 21 to be detected is in particular in the mixing chamber 5 with the associated detection partner, here the conjugate 22 incubated, so mixed or brought into contact.
  • the detection partners or conjugates 22 can be dried in the mixing chamber 5, for example, or be presented in some other way. In the former case, these are then released from the sample 3 or other liquid again.
  • the detection partners or conjugates 22 may also be added or mixed via the optionally provided detection liquid 7 of the sample 3, in particular directly into the mixing chamber 5, as already mentioned.
  • the mixture or the sample 3 is then passed into the separation chamber 10, in particular after incubation and / or for (further) incubation.
  • the binding partners or antibodies 20 are preferably already bonded or immobilized on the surface region 19.
  • the substances or analytes 21 to be detected, the analyte-conjugate complexes or the like, are then bound to the antibodies 20 and to the surface region 1, respectively.
  • unbound detection partners, conjugates 22 and in particular other, the optical investigation possible interfering substances preferably by means of the washing liquid 12 from the detection chamber 10 by appropriate rinsing of the detection chamber 10 with the Waschflüs- 12 removed.
  • the substances or analytes 21 to be detected and the detection partners or conjugates 22 to be optically detected remain bound at the surface area 19, so that now the actual optical detection or measurement can take place.
  • the immunoassay reaction described above may also be different.
  • only the analytes 21 on the antibodies 20 and the detection partners or conjugates 22 can only then bind to the already bound analyte 21.
  • the sample 3 and detection liquid 7 can be passed through the detection chamber 10 in succession, for example.
  • the antibodies 19 can optionally be occupied by different substances, such as the analyte 21 and conjugate 22 or even different conjugates 22.
  • only detection antibodies or conjugates specific to free antibodies can bind 22.
  • the term "bonding" should be understood in a very broad sense, in particular not only chemical compounds, but also any other attachment. Stick or the like. Include.
  • reaction partners such as non-bound detection partners or conjugates 22 are not removed by rinsing out of the detection chamber 10, but, for example, merely moved away from the surface region 19, for example by magnetic or electrical moving away in another area Detection chamber 10, and / or bound in or on other areas.
  • the content of the analyte 21 or of another substance in the sample 3 can thus be measured or determined.
  • the optical examination can in principle also serve other purposes, in particular any other optical detection.
  • the optical examination relates in particular to the surface area 19, wherein the optical examination particularly includes or concerns the transition to the liquid 12 or a near-surface boundary area of the liquid 12, in particular of less than 50 or 100 nm.
  • a light source 23 and an optical sensor 24 are preferably used.
  • the surface region 19 or a substance bound thereto, such as the detection partner or the conjugate 22, is irradiated with irradiated light L1 and light L2 emitted therefrom is detected by means of the sensor 24.
  • a luminescence or fluorescence measurement is carried out.
  • the angle of incidence of the incident light LI and the angle of the main detection direction for the detected light L2 to be detected are preferably different so as to mask out reflected light by means of the sensor 24.
  • the irradiation of the light LI obliquely and the detection or main detection direction of the emitted light L2 is at least substantially perpendicular to the main extension direction of the detection chamber 10 or the device 1 and / or to the flat side of the device 1 and / or to the surface extension of the surface region 19 - runs.
  • this can also be done in reverse or in any other way.
  • light LI from the UV range ie UV light, irradiated, for example with a wavelength of 250 to 400 nm, in particular of substantially 350 nm.
  • the light source 23 is preferably a UV light source.
  • the emitted or emitted light L2 or the light L2 detectable by the sensor 24 is preferably in the visible range in the illustrated example, in particular in the green range, and / or preferably has a wavelength of 500 to 650 nm, in particular substantially 550 nm.
  • the wavelengths or wavelength ranges used can vary greatly depending on the participating reaction partners, detection partners or the like and can be adapted accordingly to the respective needs.
  • the wavelength or the wavelength range of the emitted light L2 which is relevant for the detection, depends strongly on the detection partner or conjugate 22 used and can accordingly vary.
  • the detection partner or the conjugate 22 contains a lanthanide, in particular Sm, Eu or Tb.
  • a temporally resolved fluorescence measurement is particularly preferred.
  • a spectral resolution of the emitted or emitted light L2 can also take place.
  • the irradiation of the light LI and / or the detection of the emitted or radiated light L2 preferably takes place or take place through the wall forming or supporting the surface area 19, ie through the cover element 18 in the illustrated example.
  • the surface area 19 for the optical examination ie during the optical examination or fluorescence measurement, is covered by the liquid 12.
  • the liquid 12 is proposed to be optically active, in such a way that it filters, reflects, scatters, polarizes and / or absorbs light having a wavelength at least substantially corresponding to the input and / or emitted light LI or L2.
  • the optically active liquid 12 ensures, in particular, that the incident light LI can penetrate into the liquid 12 as little as possible or at least not up to an opposite surface region 26 or bottom or to the base or lower part 17, the liquid 12 can penetrate.
  • detection partners in particular adhering thereto conjugates 22 or the like.
  • Reduced or completely prevented become.
  • unwanted suggestions of the wall material in the opposite wall, which is formed here by the base or lower part 17, can be reduced or avoided altogether.
  • the optically active liquid 12 causes any existing interference signals to be blanked out.
  • interference signals is to be understood here in particular as unwanted light irradiation which can falsify or superimpose the detection or measurement of the light L2 actually emitted by the conjugates 22 on the upper surface 19 to be examined.
  • the undesired interference signals can be, for example, light radiation of the background through the often transparent base or lower part 17 into or through the liquid 12.
  • the interference signals may be emitted light which is emitted by the wall material of the base part or lower part 17 or another wall section of the detection chamber 17 or the like, in particular after appropriate excitation by the irradiated light L1.
  • the interfering signals can also be light which is emitted by detection partners or conjugates 22 remaining in the detection chamber 10-in particular also after rinsing-in particular after appropriate excitation by the irradiated light L1. In practice it is like that. even after washing or rinsing, some detection partners or conjugates 22 may be present in the liquid 12 and / or on other walls or surfaces of the detection chamber 10, such as the opposite surface or bottom region or on the base or lower part 17 or can adhere.
  • the optically active liquid 12 ensures that a significantly better signal / noise ratio can be achieved in the optical examination or fluorescence measurement.
  • unwanted spurious signals can be significantly reduced significantly.
  • the optical examination or measurement can accordingly be carried out much more accurately.
  • significantly lower contents of the analyte 21 in the sample 3 can thus be determined much more accurately.
  • a significantly improved linearity can be achieved in the detection or determination of the analyte 21 or in the realization of an immunoassay.
  • the liquid 12 in the detection chamber 10 is preferably optically activated or active by adding a dye and / or pigments or particles 25 in order to achieve the desired optical properties, as described above.
  • the dye, pigment or particle 25 can also be any mixture or mixture of different substances.
  • pigments and / or particles in particular with an average diameter of about 10-30 nm and / or with or consisting of titanium dioxide or the like.
  • titanium dioxide nanoparticles (E 171) or other nanoparticles it is possible, for example, to achieve absorption in the UV range, wherein the liquid 12 can remain transparent even in the visible range.
  • the optically active liquid 32 with the dye is completely or at least substantially, in particular more than 50%. saturated.
  • the surface region 19 to be investigated is of the optically active liquid 12 with a layer thickness D (in Fig. 3 indicated) of preferably more than 0, 1 .mu.m, advantageously more than 1 .mu.m, in particular more than 10 .mu.m, preferably more than 50 microns , Particularly preferably about 100 microns or more, covered, in order to ensure a sufficiently strong optical effect of the liquid 12 can.
  • the desired optical effect of the liquid 12 can be enhanced by other particles or substances therein, which provide, for example, for a scattering, reflection or refraction of the light, so that optionally a correspondingly lower concentration of the dye or of the particles / pigments 25 and / or.
  • the optically active liquid 12 is preferably used as an optically neutral background for the detection of the emitted or emitted light L2, in particular for a fluorescence measurement.
  • the optically active liquid 12 is preferably used as an optical filter for light behind the surface region 19, ie on the side facing away from the detection or measuring side.
  • the optically active liquid 12 may, in particular, be a washing liquid 12 provided for the immunoassay reaction or another reaction.
  • the optically active liquid 12 may in principle also be any other reaction liquid which is used in particular for the formation and / or binding of the substance and / or an immunoassay reaction.
  • the optically active liquid 12 can also be a liquid which is additionally used independently of the reaction or binding of a substance to be detected or the like, which liquid is optionally used only or additionally for the purpose of optical examination or in the detectors. onshunt 10 is initiated.
  • corresponding dyes, pigments and / or particles 25 may also be added to or dissolved in the detection chamber 10 of the liquid 12.
  • the dyes, pigments and / or particles 25 may be present in the detection chamber 10 in a dried form and dissolved or added in any other way.
  • the optically active liquid 12 is layered with an optically non-active liquid.
  • the optically active liquid layer preferably directly adjoins the surface region 19.
  • an optical non-active liquid layer between the optical active liquid layer and the surface area 19 to be examined lie.
  • optical properties of the optically active liquid 12 and / or the concentration of the dyes, pigments and / or particles 25 it is fundamentally possible for the optical properties of the optically active liquid 12 and / or the concentration of the dyes, pigments and / or particles 25 to vary spatially, in particular over the thickness D of the detection chamber 10 or perpendicular to the surface region 19.
  • the optical properties of the optically active liquid 12 can also be selectively varied in order, for example, to be able to selectively detect or measure radiated light L2 from different spatial regions and / or in different wavelength ranges.
  • a dye, pigment or particle 25 may additionally be added in order to vary the optical properties of the optically active liquid 12 in a desired manner, or the optically active liquid 12 against a optically inactive fluid or vice versa.
  • the optically active liquid 12 can be used to selectively (optically) selectively cover, in particular, an opposing (additional) surface region 26, to irradiate the surface region 26 or a substance disposed thereon and / or to selectively prevent or at least minimize emission thereof.
  • the optical examination or measurement shown in FIG. 3 is carried out in particular in addition to (before or after) the optical examination of the surface area 19 shown in FIG. 2.
  • the present in the detection chamber 10 during the optical examination Liquid 12 namely not optically active.
  • the optically active liquid 12 shown in FIG. 2 for example, can be replaced by a non-optically active liquid 12 as shown in FIG. 3 or vice versa.
  • the additional surface region 26, which in particular faces the surface region 19 and is separated therefrom by the liquid 12 can likewise be examined optically.
  • 3 schematically indicates that an additional analyte 28 may be bound to an additional conjugate 29 on the additional surface area 26, for example by means of additional antibody 27.
  • the additional antibodies 27 are then immobilized on the additional surface area 26, for example.
  • the previous explanations regarding a preferably proceeding immunoassay reaction or other reaction for binding the additional analyte 28 or the additional, optically detectable detection partner or conjugate 29 apply in particular accordingly.
  • the irradiated light L1 can penetrate through the liquid 12 up to the additional surface region 26 and excite there the additional conjugates 29. Accordingly, there is then an additional emission of light L3, which is additionally detectable by means of the sensor 24 or with the fluorescence measurement, as indicated in FIG. 3. In this case, a sum signal then results, for example, from the light L2 emitted by the first surface region 19 and the light L3 emitted by the second or additional surface region 26. If before or after the measurement is carried out only for the (first) surface area 19 as described with reference to FIG. 2, the presence of the additional conjugate 29 and thus of the additional analyte 28 can be qualitatively or quantitatively determined by appropriate subtraction.
  • optically active liquid 12 can in particular suppress or hide intrinsic or autofluorescent properties of wall material or other material and / or background influences. Furthermore, the optically active liquid 12 can reduce or substantially avoid negative influences of non-specific bonds-such as binding of the conjugate 22/29 independently of the associated antibody 20/27 on other binding partners or surface areas-in optical examinations or measuring methods. It should be noted that the optically active liquid can also act or be used as a so-called quencher or cut-off filter. Through the so-called “quenching" can prevent that Fluofore pass into the excited state or that excited Fluofore be transferred without radiation in the ground state.
  • liquids 12 with different optical properties ie, for example, an optically active liquid 12 and / or, for example, different optically active liquids 12, optionally one after the other in the detection chamber 10 or different areas of the detection 10 or via various surface areas 19, 26 to be examined.
  • the present invention or the optically active liquid 12 can also be used when light LI is excited, but the signal of the sample 3 or the analyte 21/28 or the detection partner or conjugate 22/29 to be examined is not optically, but in another way, for example by an electrical measurement or resistance change, a photothermal method or the like, is detected.
  • a reaction is started only by irradiation of the light LI, for example. The proof is then in particular not optically.

Abstract

L'invention concerne un procédé et un dispositif permettant l'examen optique d'une zone superficielle. La teneur en une substance liée à la zone superficielle est déterminée par mesure de fluorescence. La lumière parasite est supprimée par le fait que cette zone superficielle est recouverte d'un liquide optiquement actif qui filtre, réfléchit, diffuse et/ou absorbe la lumière dont la longueur d'onde correspond au moins sensiblement à celle de la lumière incidente et/ou émise.
EP11721538.4A 2010-05-31 2011-05-26 Procédé et dispositif d'examen optique Withdrawn EP2577311A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP11721538.4A EP2577311A1 (fr) 2010-05-31 2011-05-26 Procédé et dispositif d'examen optique

Applications Claiming Priority (3)

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EP10005631 2010-05-31
PCT/EP2011/058635 WO2011151250A1 (fr) 2010-05-31 2011-05-26 Procédé et dispositif d'examen optique
EP11721538.4A EP2577311A1 (fr) 2010-05-31 2011-05-26 Procédé et dispositif d'examen optique

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EP (1) EP2577311A1 (fr)
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JP2015500500A (ja) * 2011-12-15 2015-01-05 インストゥルメンテーション ラボラトリー カンパニー 使い捨てマイクロ流体デバイスにおける蛍光測定及びその方法
WO2016062788A1 (fr) 2014-10-24 2016-04-28 Ait Austrian Institute Of Technology Gmbh Puce microfluidique pour analyse biologique
JP6825579B2 (ja) * 2015-12-28 2021-02-03 凸版印刷株式会社 マイクロ流体デバイスおよび観察方法
JP6808940B2 (ja) * 2016-01-29 2021-01-06 凸版印刷株式会社 アレイデバイス、生体分子解析用キット及び溶媒の封止方法

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US20120208292A1 (en) 2012-08-16
WO2011151250A1 (fr) 2011-12-08
US20130236985A1 (en) 2013-09-12
JP5822168B2 (ja) 2015-11-24
JP2013528805A (ja) 2013-07-11

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