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EP1685900B1 - Use of a device for analysing a liquid sample - Google Patents

Use of a device for analysing a liquid sample Download PDF

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Publication number
EP1685900B1
EP1685900B1 EP06001069A EP06001069A EP1685900B1 EP 1685900 B1 EP1685900 B1 EP 1685900B1 EP 06001069 A EP06001069 A EP 06001069A EP 06001069 A EP06001069 A EP 06001069A EP 1685900 B1 EP1685900 B1 EP 1685900B1
Authority
EP
European Patent Office
Prior art keywords
sample liquid
reaction
channel
reagent
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.)
Active
Application number
EP06001069A
Other languages
German (de)
French (fr)
Other versions
EP1685900A1 (en
Inventor
Gert Dr. Blankenstein
Ralf-Peter Dr. Peters
Thomas Willms
Claus Marquordt
Christian Schön
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
Priority claimed from DE200510003961 external-priority patent/DE102005003961A1/en
Application filed by Boehringer Ingelheim Microparts GmbH filed Critical Boehringer Ingelheim Microparts GmbH
Publication of EP1685900A1 publication Critical patent/EP1685900A1/en
Application granted granted Critical
Publication of EP1685900B1 publication Critical patent/EP1685900B1/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • B01L3/502738Containers 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 integrated valves
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • B01L3/50273Containers 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 the means or forces applied to move the fluids
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06Fluid handling related problems
    • B01L2200/0605Metering of fluids
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06Fluid handling related problems
    • B01L2200/0684Venting, avoiding backpressure, avoid gas bubbles
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/16Reagents, handling or storing thereof
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0809Geometry, shape and general structure rectangular shaped
    • B01L2300/0825Test strips
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/16Surface properties and coatings
    • B01L2300/161Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
    • B01L2300/165Specific details about hydrophobic, oleophobic surfaces
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0403Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406Moving fluids with specific forces or mechanical means specific forces capillary forces
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/06Valves, specific forms thereof
    • B01L2400/0688Valves, specific forms thereof surface tension valves, capillary stop, capillary break
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/08Regulating or influencing the flow resistance
    • B01L2400/084Passive control of flow resistance
    • B01L2400/086Passive control of flow resistance using baffles or other fixed flow obstructions

Definitions

  • the present invention relates to an apparatus for assaying sample liquid such as blood, blood plasma, urine, saliva or the like.
  • the present invention is concerned with microfluidic systems.
  • the following statements relate to devices in which act capillary forces and are particularly crucial for the function.
  • test filter strips are frequently used which consist of paper, foils, filters, membranes or the like. consist.
  • Such a test filter strip is designed for sample application and also assumes transport functions. For example, the sample liquid is transported in the test filter strip due to capillary forces in a non-woven material.
  • the sample liquid can react with previously introduced reagents and, for example, cause a color change upon detection of an analyte in the sample liquid.
  • test filter strips allow only a relatively inaccurate, qualitative detection of an analyte.
  • microcapillary systems are known for examining a sample fluid.
  • the EP 1 201 304 A2 discloses a microstructured platform for examining a sample fluid.
  • the platform has a filling area, an examination area and a channel system.
  • the sample liquid can be absorbed and promoted solely by capillary forces.
  • the known platform has delay structures along the edges of a broad, flat channel, in particular in the examination region.
  • the sample liquid can be stopped in the known platform, if necessary, at a predetermined location for a predetermined period, for example, a chemical reaction or physical process such as heating or cooling.
  • the EP 1 201 304 A2 does not deal with as accurate a study as possible, in particular quantitative analysis, of a sample liquid.
  • the US2004 / 0077103 A1 describes a device and a method in which a sample first dissolves a reagent and transferred after a predetermined period of time in an examination area.
  • the older post-published WO2005 / 119211 A1 discloses a device with a side wall-free channel, wherein a channel portion may form a sample liquid sample area.
  • the design of the channel is chosen so that an optimal ventilation of the channel takes place when filling the channel with sample liquid W02005 / 119211 A1 also does not deal with a quantitative study of a sample liquid.
  • the present invention has for its object to provide a use of a device for examining sample liquid, such as blood, blood plasma, urine, saliva or the like., Which at low cost, preferably a quantitative examination, in particular determination of blood sugar, blood fat, enzymes or other values.
  • sample liquid such as blood, blood plasma, urine, saliva or the like.
  • a substantially more accurate examination of a sample liquid in particular a quantitative determination of at least one analyte in the sample footage, is made possible.
  • the reaction volume with the dissolved reagent or reaction products from the reaction area in the study area further promoted, the lateral, at least substantially laminar flow and / or the at least substantially rectilinear flow front of the sample liquid leads to a low dispersion, so at least substantially uniform Concentration profile of the dissolved reagent or reaction products can be reached in the further funded in the study area reaction volume. Accordingly, within a defined time, a much more precise investigation, in particular determination of values in the aforementioned sense, can take place.
  • complexes or compounds formed by the dissolved reagent and an analyte to be determined can be bound in the examination area by means of an immobilized detection chemical and subsequently - for example optically - confirmed or measured. From this, it is then possible to determine, for example, the concentration of the analyte in the sample liquid.
  • a further, independently realizable aspect of the present invention is to provide the reaction area, the examination area and / or the channel with an at least substantially constant cross-section and / or the height of the channel at least by a factor of 10 smaller than the width of the channel form and / or the reaction area and / or the examination area maximum as long as wide or shorter form.
  • the above measures are conducive to quantitative analysis or biochemical investigations.
  • a defined examination of the defined reaction volume is made possible or an undesirable dispersion of the reaction volume is avoided.
  • Further advantages include short examination times, fast reactions, short diffusion paths and / or short flow paths.
  • Fig. 1 shows in a schematic section a part of a first embodiment of a proposed device 1 for examining a sample liquid 2, in particular blood plasma o. The like.
  • the device 1 has a channel 3 receiving the pump fluid 2 by capillary forces.
  • the channel 3 is preferably bounded or formed by only two opposite, in particular substantially flat surfaces or flat sides 4 and 5.
  • the device 1 has a preferably plate-shaped support 6 and an associated cover 7, between which the channel 3 is formed.
  • the support 6 is excluded for the formation of the required microstructures and the cover 7 flat, preferably at least substantially ausappelungsoko formed.
  • this can be the other way around.
  • both the carrier 6 and the cover 7 can be excluded and / or formed with projections for forming the desired structures and, if necessary, for holding chemicals, reagents, examination devices or the like, not shown.
  • the device is thus a so-called microchip (platform with microstructure).
  • Fig. 2 shows in a schematic plan view of the carrier 6 of the device 1, but without cover 7 and without sample liquid 2.
  • the channel 3 preferably has one inlet immediately following, in the illustrated example in particular with a trench 8, a reaction area 9, an examination area 10 and / or a Collection area 11 on.
  • the device 1 preferably has only a single channel 3.
  • the channel 3 is to be understood in terms of a single capillary. If necessary, however, the channel 3 can lead in different directions or to different areas or branch.
  • the sample liquid 2 preferably flows exclusively through capillary forces into or into the channel 3 in the flow direction S, as in FIG Fig. 1 indicated.
  • the sample liquid 2 may additionally or alternatively also be conveyed, for example, by pressure through the channel 3.
  • the channel 3 preferably has a substantially rectangular and / or flat cross section transverse to the flow direction S of the sample liquid 2.
  • the distance of the channel 3 bounding, preferably parallel surfaces 4 and 5 - is at most 2000 microns, preferably at most 500 .mu.m, in particular about 50 to 200 microns.
  • the width of the channel 3 is preferably about 100 to 5000 microns, in particular about 200 to 4000 microns.
  • the height H of the channel 3 is substantially smaller, in particular at least by a factor of 10 or 100, than the width of the channel 3.
  • the absorption volume of the channel 3 is preferably less than 1 ml, in particular less than 100 ⁇ l, particularly preferably not more than 10 ⁇ l.
  • the device 1 thus forms a microfluidic system.
  • the device 1 is used for microfluidic diagnostics for medical or non-medical purposes or other examinations.
  • the channel 3 and its main extension plane preferably extend at least substantially horizontally in the position of use. Depending on the intended use or constructive solution, however, a different orientation is possible, especially since the reception or filling of the channel 3 with sample liquid 2 is preferably determined or effected at least primarily by capillary forces.
  • reaction region 9 with a reagent which can be dissolved and / or reacted with the sample liquid 2 and the preferably immediately adjoining examination region 10 are formed in the channel 3, preferably successively on the same flat side 4 of the channel 3.
  • the reaction region 9 has a reagent, which is preferably detachable from the sample liquid 2, for an analyte to be determined in the sample liquid 2.
  • the reagent is antibodies directed against the analyte of interest and bound to indicators (dyes, colorant particles, e.g., colloidal gold).
  • the reagent is dissolved when filled with the sample liquid 2.
  • the analyte, when contained in the sample fluid 2 then reacts with the antibody bound to the dye, and in particular forms a compound or complex.
  • the reagent reacts with the analyte and in particular forms a reaction product, even if the reagent is not dissolved if necessary.
  • the following statements with regard to the (dissolved) reagent therefore apply correspondingly to the reaction product.
  • the examination region 10 is provided with a preferably immobilized detection chemical, which in particular binds compounds or complexes of analyte and reagent or the reaction product. Unbound reagent and other ingredients then flow with the sample fluid 2 further into the collection area 11, where they are taken up, thereby preventing backflow.
  • the bound reagent can be determined optically and from this the presence and in particular the concentration of the analyte in the sample liquid 2 can be determined.
  • a particularly quantitative examination of the sample liquid 2 is made possible.
  • the device 1 has a device 12 for temporary stopping of the sample foot 2 in the reaction area 9 for dissolving and / or reacting the reagent and / or in the examination area 10
  • the device 12 is preferably designed such that the temporary stop by the sample liquid 2 itself, such as in the EP 1 441 131 Al described, or by a control fluid, not shown, or by a selective vent, such as in the EP 1419 818 Al described, fixed or can be canceled.
  • the device 12 holds the sample liquid 2 for a predetermined period of time, possibly only after the complete filling of the reaction zone 9, and / or until the reaction zone 9 has been completely filled.
  • the device 12 has, in particular, a control channel 13 which supplies sample liquid 2 to a liquid stop 14 arranged between the reaction zone 9 and the examination zone 10 within a defined time, so that then the sample liquid 2 or reaction volume located in the reaction zone 9 on sample liquid 2 overcome the liquid stop 14 and can flow into the examination area 10.
  • the device 1 has a further device 12 for temporary stopping of the sample liquid 2 - in particular the reaction volume of sample liquid 2 which has previously flowed from the reaction region 9 into the examination region 10 and contains dissolved reagent or the reaction product - in the examination region 10 in order to obtain the most accurate possible or to permit quantitative determination, in particular in order to enable an at least substantially complete reaction or binding of the compounds or complexes of reagent and analyte or of the reaction product to the detection chemical in the examination area 10.
  • the further device 12 is designed in particular according to the aforementioned device 12. Accordingly, in turn, a control channel 13 is provided, the sample liquid 2 after or in a defined time to a arranged between the examination area 10 and the downstream collection area 11 liquid stop 14 supplies, so that then the sample liquid 2 or located in the examination area 10 reaction volume of sample liquid. 2 overcome the liquid stop 14 and continue to flow into the collection area 11 and the inflowing sample liquid. 2 then a washing out of unbound reagent or reaction product in the examination area 10 can cause.
  • the liquid stop 14 is formed in particular by a groove-like or trench-like depression transverse to the flow direction S.
  • other constructive solutions are possible.
  • Fig. 3 shows in one too Fig. 2 corresponding plan view of the unfilled carrier 6 of a device 1 according to a second embodiment.
  • the devices 12 each have here, instead of a preferably groove-like or trench-like liquid stop 14, at least one preferably web-like barrier 15, in particular two or more barriers 15 arranged one behind the other. Thus, if necessary, a corresponding temporary stopping of the sample liquid 2 can be achieved.
  • the channel 3 does not have a substantially constant cross-section, in contrast to the first embodiment. Instead, the cross section of the channel 3 at the transition from the reaction region 9 to the examination region 10 and / or during the transition from the examination region 10 into the collection region 11 is reduced. This reduction in cross-section is preferably achieved by uniform tapering of the liquid stream and subsequent spreading of the liquid stream.
  • the device 12 or barrier 15 is then preferably arranged in the region of the reduced cross section.
  • Said cross-sectional reduction already leads to a reduction in the volume flow through the channel 3, so that, if necessary, no complete temporary stop is required.
  • a delay caused by the barriers 15 of the flow or reduction of the volume flow may be sufficient.
  • the device 1 further comprises a means 16 for preventing the lateral advancement of sample liquid 2 in the flow direction S and / or for generating an in Fig. 1 indicated, according to the plan view Fig. 2 little curved or rectilinear flow front F of the sample liquid 2 or to produce a homogeneous or laminar flow.
  • the means 15 is formed in that the channel 3 is formed open at least on the long side.
  • the side of the channel 3 is followed by a recess 17, which is formed in particular nut- or trench-like.
  • a lateral liquid stop for the sample liquid 2 - that is, a flow impediment that can not be overcome by capillary forces - is formed and the sample liquid 2 is guided along the open longitudinal sides in the channel 3 without sidewalls.
  • the recess 17 preferably connects sharp-edged to the channel 3, as in Fig. 1 . 3 and 4 indicated.
  • the recess 17 is formed only in the carrier 6, extends in the illustration according to Fig, 1 . 3 and 4 Thus, essentially only downwardly with respect to a lateral projection of the channel 3.
  • the recess 17 may, however, optionally also upwards or on both sides of the lateral projection of the channel 3, so in particular extend upwards and downwards.
  • the preferably rectangular cross-section recess 17 leads to such, in particular stepped or sudden cross-sectional enlargement that reduce the capillary forces such that said liquid stop for the sample liquid 2 is formed in the transition from channel 3 to the recess 17 out.
  • the height of the recess 17 is at least twice as large as the height H of the channel 3.
  • the recess 17 extends in the representation example along the open side of the channel 3, in particular, it is formed circumferentially around the channel 3 open on all sides.
  • a corresponding side wallless leadership of the sample liquid 2 in the channel 3 is also in the in Fig. 5 illustrated, third embodiment of the device 1 through the lateral recess 17 is possible.
  • the sample liquid 2 is guided here only on a bottom or flat side 4.
  • the sample liquid 2 is thus not in contact with an opposite flat side 5 as in the first embodiment.
  • the cover 7 is excluded accordingly or the surface 4 is arranged correspondingly deep in the carrier 6 in order to maintain a sufficient distance to the then possibly flat cover 7 can.
  • the thickness of the liquid film formed by the sample liquid 2 on the surface 4 depends in particular on the wetting behavior and on the quantity of sample liquid 2 fed in, in particular metered in. The corresponding dimensions then apply to the liquid film, as explained for the channel 3 in the first embodiment.
  • Fig. 6 shows in a schematic section a fourth embodiment of the proposed device 1, wherein a side portion of the channel 3 is shown enlarged for clarity.
  • the means 16 for preventing the lateral advancement of sample liquid 2, for producing a slightly curved or rectilinear flow front F and / or for generating a homogeneous or laminar flow may alternatively or additionally also have a side wall 18 which delimits the channel 3 on the long side or on all sides by forming corresponding guide elements or delay structures, in particular projections or elevations 19 or the like.
  • the flow or filling speed along the side wall 18 in the flow direction S reduces the filling speed, in particular so that the filling rate of the sample liquid 2 at the edge that in the central region of Channel 3 does not exceed, but this at least substantially corresponds.
  • the wetting of the side wall 18 can also be modified, in particular reduced, in such a way that the undesirable advancing of sample liquid 2 along the side wall 18 is avoided.
  • the channel 3 has at least one guide element for influencing, in particular equalizing, the filling with the sample liquid 2.
  • the channel 3 preferably has regularly distributed elevations 19 as guide elements on the flat side 4 or possibly both flat sides 4, 5, as in FIG Fig. 1, 2 and 4 to 6 shown. These are in particular arranged in rows transversely, preferably vertically, or longitudinally to the flow direction S, in particular alternately transversely offset.
  • the means 16 also includes, if necessary, said guide elements.
  • the area density, the distance and / or the size of the elevations 19 vary, in particular as a function of the respective distance to the inlet, in order to achieve a desired course of the capillary forces or possibly a compensation of flow resistances.
  • the elevations 19 are preferably web-like, hump-shaped or column-like, in particular with a round or polygonal base surface. Alternatively or additionally, however, it is also possible to provide depressions, such as the trench 8, or the barriers 15 or other guide elements, which extend transversely or longitudinally to the flow direction S of the channel 3.
  • the preferably provided groove-like, in cross-section in particular rectangular or semicircular trench 8 has a much smaller depth than the liquid stop 14 and the recess 17 and therefore forms only a temporary liquid stop to equalize the liquid front F.
  • the Proben thoroughlylcezt 2 only after filling the channel 3 over the entire cross section fills the trench 8 and then the subsequent channel region.
  • a highly uniform filling of the channel 3 in particular by capillary forces with at least substantially straight-line or perpendicular to the flow direction S extending liquid front F can be achieved by the combination of side wallless guidance of the sample liquid 2 and the guide elements.
  • the channel 3 can be formed partially or even at least substantially smoothly or even, that is to say in particular without guide elements, as in FIG Fig. 3 indicated.
  • the structuring or texturing of the reaction area 9 and / or the examination area 10, in particular by guide elements, such as the elevations 19 or the like, facilitates the preferably uniform application of a chemical or the like, which subsequently dries and thereby, for example, a dry chemical or immobilized Chemical forms.
  • the device 1 preferably has a vent 20 in communication with the recess 17, as in FIG Fig. 4 indicated. This allows in a very simple way effective ventilation. This is conducive to a uniform, bubble-free filling of the channel 3 with the sample liquid 2.
  • the sample liquid 2 is conducted into the reaction zone 9 by capillary forces in the channel 3.
  • the sample liquid 2 flows in the channel 3 - at least in the reaction region 9 and examination region 10 - laterally over the flat side 4 and preferably at least substantially laminar or with homogeneous flow velocity or slightly curved or rectilinear flow front F.
  • said means 16 - In particular in combination with the preferably at least in the reaction region 9 and / or examination area 10 provided guide elements - achieved.
  • the sample liquid 2 is temporarily stopped by the device 12 for a preferably predetermined time.
  • the sample liquid 2 can dissolve or react with the reagent, which is preferably present as a dry chemical, for determining an analyte in the sample liquid 2.
  • the reagent is, for example a conjugate composed of an antibody binding to the analyte and a dye particle or the like. is formed. The dissolved reagent or conjugate then binds to the analyte.
  • the reaction area 9 is filled with a defined reaction volume of the sample foot 2, so that the reagent dissolves at least substantially only in this reaction volume or only reacts with it.
  • the temporary stop an undesired dispersion or extensive distribution of the reagent or a reaction product of the reagent in the sample liquid 2 can thus already be avoided.
  • the time for the temporary stop is preferably chosen such that the reagent is dissolved to at least 90%, in particular at least 95% or substantially more in particular in the reaction volume of sample liquid 2 or reacts with it. If necessary, the dissolution or reaction by heat or other measures, such as applying a voltage or the like, Can be supported.
  • the reagent is preferably applied uniformly or in a predetermined concentration distribution on the flat side 4 in the reaction region 9.
  • the most even distribution in the reaction volume is also beneficial that the reagent on the flat side 4 of the channel 3 is arranged and that due to the relatively low channel height H accordingly a fast diffusion and thus uniform distribution of the dissolved reagent or the reaction product in the reaction volume can be achieved ,
  • the device 12 releases the sample liquid 2, so that the sample liquid 2 - in particular the defined reaction volume - from the reaction region 9 in the study area 10 can continue to flow. Due to the measures mentioned, and in particular the means 16, a particularly small dispersion of the dissolved reagent as well as the compounds or complexes formed from the reagent and the analyte can in turn be prevented from the reaction volume. In particular, it is achieved that the reagent or the reaction product to at least 90%, preferably 95% or more flows together with the reaction volume in the examination area 10 or is promoted.
  • the sample liquid 2 or the reaction volume is again temporarily stopped, if necessary, for the respectively desired examination, in particular the binding or compounds formed from the reagent and the analyte in the examination area 10, preferably on the flat side 4 immobilized detection chemical support.
  • a temporary stop in the examination area 10 is not absolutely necessary, so that the further, the examination area 10 associated device 12 may possibly be omitted.
  • the intended detection chemical is, in particular, an immobilized dry chemical, for example a capture antibody, which captures and binds the compounds or complexes of reagent and analyte.
  • further examination steps can be carried out by means of other further chemicals in the examination area 10 or in a plurality of examination areas 10 arranged one after the other. For example, an investigation can also be made as to whether the analyte to be determined is contained in the sample liquid 2 at all.
  • the examination region 10 preferably directly adjoins the reaction region 9, so that undesired dispersion of the reagent or of compounds or complexes formed by the reagent with the analyte or of other reaction products from the reaction volume into other regions of the sample liquid 2 is at least substantially prevented or minimized. Due to the fact that the examination area 10 preferably immediately adjacent to the reaction region 9, namely the dead volume and thus the dispersion are minimized.
  • the reaction area 9 and / or the examination area 10 can be formed relatively short in the flow direction S, so that a total of very short flow paths and thus a low dispersion can be achieved.
  • the reaction area 9 and / or the examination area 10 is formed only as long as or shorter than the width of the channel 3.
  • the dispersion which can be achieved according to the proposal in the examination zone 10 is so small that the concentration of the reagent or reaction product in the reaction volume in the examination zone 10 varies by a maximum of 10%, preferably less than 5%, very preferably at most 3%.
  • the sample liquid 2 flows further into the collecting area 11, which may optionally be provided with an absorbent material and / or guide elements, such as the elevations 19, to absorb the sample liquid 2 and to prevent a reflux
  • the volume of Collection area 11 is preferably greater than the reaction volume by at least a factor of 2 or 5 in order to achieve efficient washing out of unbound reagent, unbound reaction products and / or other possibly interfering particles or substances from examination area 10.
  • the determination of the bound in the examination area 10 reagent or reaction product is preferably carried out optically, for example spectroscopically.
  • the Reagent or reaction product for example, as a conjugate of an antibody and a color complex, color particles or the like. is constructed.
  • the concentration of the analyte in the sample liquid 2 can then be determined from the number or concentration of bound complexes or compounds. Consequently, the proposed device 1 and the method described above allow a comparison with conventional test filter strips or the like. much more accurate examination of the sample liquid 2, in particular quantitative determination of an analyte or optionally also several analytes in the sample liquid 2.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

A device for testing sample liquids (2) with a reaction zone (9) with a reagent (R) on the flat side, a test zone (10) and a device for holding (2) in zone (9) for the dissolution and/or reaction of (R), in which there is a means of producing a straight flow front in (2), and at least 90% of (R) reacts with (2) and is transferrable with it to zone (10), or the concentration of (R) or reaction product in (10) varies by not more than 10%. A device (1) for testing sample liquids (2), with: (a) a channel (3) for taking up and transporting the liquid (2) by means of capillary forces, which has a flat side (4) over which the liquid (2) passes with lateral and/or laminar flow; (b) a reaction zone (9) with a soluble and/or reactive reagent (R) on the flat side (4), which can be completely filled with (2) to define a reaction volume (V) of liquid (2); (c) a test zone (10) formed by the channel (3) and following zone (9); and (d) a device (12) for keeping (2) in the reaction zone (9) temporarily for the dissolution and/or reaction of (R). The device (1) also has a means (16) of producing a minimally curved or straight flow front (F) in the liquid (2); the reagent (R) is soluble in and/or reacts with the defined volume (V) of (2) to the extent of at least 90%, and at least 90% of the dissolved (R) or a reaction product of (R) is transferrable to zone (10) together with (V), or the concentration of dissolved (R) or reaction product within (V) in zone (10) varies by not more than 10%. An independent claim is also included for a method for testing sample liquids (2) with the above device, in which the liquid (2) flows from zone (9) into zone (10) with an at least substantially straight flow front (F) and/or at least without any substantial change in flow cross-section.

Description

Die vorliegende Erfindung betrifft eine Vorrichtung zur Untersuchung von Probenflüssigkeit, wie Blut, Blutplasma, Urin, Speichel o.dgl.The present invention relates to an apparatus for assaying sample liquid such as blood, blood plasma, urine, saliva or the like.

Die vorliegende Erfindung befaßt sich mit mikrofluidischen Systemen bzw. Vorrichtungen. Die nachfolgenden Ausführungen beziehen sich auf Vorrichtungen, bei denen Kapillarkräfte wirken und insbesondere für die Funktion entscheidend sind.The present invention is concerned with microfluidic systems. The following statements relate to devices in which act capillary forces and are particularly crucial for the function.

Für die Untersuchung einer Probenflüssigkeit insbesondere zur Bestimmung von Blutzucker, Blutfett, Enzymen oder sonstigen Werten, werden häufig sogenannte Testfilterstreifen verwendet, die aus Papier, Folien, Filter, Membranen o.dgl. bestehen. Ein derartiger Testfilterstreifen ist zur Probenaufgabe ausgebildet und übernimmt auch Transportfiunktionen. Beispielsweise wird die Probenflüssigkeit aufgrund von Kapillarkräften in einem vliesartigen Material im Testfilterstreifen transportiert. Die Probenflüssigkeit kann dabei mit zuvor eingebrachten Reagenzien reagieren und beispielsweise einen Farbumschlag bei Detektion eines Analyten in der Probenflüssigkeit bewirken. Derartige Testfilterstreifen gestatten jedoch nur eine verhältnismäßig ungenaue, qualitative Detektion eines Analyten.For the examination of a sample liquid, in particular for the determination of blood sugar, blood fat, enzymes or other values, so-called test filter strips are frequently used which consist of paper, foils, filters, membranes or the like. consist. Such a test filter strip is designed for sample application and also assumes transport functions. For example, the sample liquid is transported in the test filter strip due to capillary forces in a non-woven material. The sample liquid can react with previously introduced reagents and, for example, cause a color change upon detection of an analyte in the sample liquid. However, such test filter strips allow only a relatively inaccurate, qualitative detection of an analyte.

Alternativ sind Mikrokapillarsysteme für die Untersuchung einer Probenflüssigkeit bekannt. Die EP 1 201 304 A2 offenbart z.B. eine mikrostrukturierte Plattform für die Untersuchung einer Probenflüssigkeit. Die Plattform weist einen Einfüllbereich, einen Untersuchungsbereich und ein Kanalsystem auf. Die Probenflüssigkeit kann allein durch Kapillarkräfte aufgenommen und gefördert werden. Um eine möglichst wenig gekrümmte Strömungsfront und eine homogene Strömungsgeschwindigkeit zu bewirken, weist die bekannte Plattform Verzögerungsstrukturen entlang der Ränder eines breiten, flachen Kanals auf, insbesondere im Untersuchungsbereich. Weiter kann die Probenflüssigkeit bei der bekannten Plattform bedarfsweise an einer vorgegebenen Stelle für eine vorgegebene Dauer angehalten werden, um beispielsweise eine chemische Reaktion oder einen physikalischen Vorgang, wie Erwärmen oder Kühlen, zu ermöglichen. Die EP 1 201 304 A2 befasst sich jedoch nicht mit einer möglichst genauen Untersuchung, insbesondere quantitativen Untersuchung, einer Probenflüssigkeit.Alternatively, microcapillary systems are known for examining a sample fluid. The EP 1 201 304 A2 For example, discloses a microstructured platform for examining a sample fluid. The platform has a filling area, an examination area and a channel system. The sample liquid can be absorbed and promoted solely by capillary forces. In order to achieve the least possible curved flow front and a homogeneous flow velocity, the known platform has delay structures along the edges of a broad, flat channel, in particular in the examination region. Further, the sample liquid can be stopped in the known platform, if necessary, at a predetermined location for a predetermined period, for example, a chemical reaction or physical process such as heating or cooling. The EP 1 201 304 A2 However, it does not deal with as accurate a study as possible, in particular quantitative analysis, of a sample liquid.

Die US2004/0077103 A1 beschreibt eine Vorrichtung und ein Verfahren, bei welchem eine Probe zunächst ein Reagenz löst und nach einer vorbestimmten Zeitdauer in einen Untersuchungsbereich überführt.The US2004 / 0077103 A1 describes a device and a method in which a sample first dissolves a reagent and transferred after a predetermined period of time in an examination area.

Die ältere nachveröffentlichte WO2005/119211 A1 offenbart eine Vorrichtung mit einem seitenwandfreien Kanal, wobei ein Kanalabschnitt einen Untersuchungsbereich für Probenflüssigkeit bilden kann. Die Ausgestaltung des Kanals ist so gewählt, dass eine optimale Entlüftung des Kanals bei der Befüllung des Kanals mit Probenflüssigkeit erfolgt- Die W02005/119211 A1 befasst sich ebenfalls nicht mit einer quantitativen Untersuchung einer Probenflüssigkeit.The older post-published WO2005 / 119211 A1 discloses a device with a side wall-free channel, wherein a channel portion may form a sample liquid sample area. The design of the channel is chosen so that an optimal ventilation of the channel takes place when filling the channel with sample liquid W02005 / 119211 A1 also does not deal with a quantitative study of a sample liquid.

Der vorliegenden Erfindung liegt die Aufgabe zugrunde, eine Verwendung einer Vorrichtung zur Untersuchung von Probenflüssigkeit, wie Blut, Blutplasma, Urin, Speichel o.dgl., anzugeben, welche bei geringem Aufwand eine vorzugsweise quantitative Untersuchung, insbesondere Bestimmung von Blutzucker, Blutfett, Enzymen oder sonstigen Werten, ermöglicht.The present invention has for its object to provide a use of a device for examining sample liquid, such as blood, blood plasma, urine, saliva or the like., Which at low cost, preferably a quantitative examination, in particular determination of blood sugar, blood fat, enzymes or other values.

Die obige Aufgabe wird durch die Verwendung einer Vorrichtung gemäß Anspruch 1 gelöst. Vorteilhafte Weiterbildungen sind Gegenstand der Unteransprüche 2 bis 4.The above object is achieved by the use of a device according to claim 1. Advantageous developments are subject of the dependent claims 2 to 4.

Mittels der vorschlagsgemäßen Kombination von Maßnahmen wird eine wesentlich genauere Untersuchung einer Probenflüssigkeit, insbesondere eine quantitative Bestimmung mindestens eines Analyten in der Probenfüßigkeit, ermöglicht. Durch das definierte Anhalten der Probenflüssigkeit im Reaktionsbereich wird ein Reaktionsvolumen festgelegt, in dem ein Reagenz im Reaktionsbereich definiert lösbar ist bzw. mit dem das Reagenz reagieren kann. Anschließend wird das Reaktionsvolumen mit dem gelösten Reagenz oder Reaktionsprodukten vom Reaktionsbereich in den Untersuchungsbereich weiter gefördert, wobei die laterale, zumindest im wesentlichen laminare Strömung und/oder die zumindest im wesentlichen geradlinige Strömungsfront der Probenflüssigkeit zu einer geringen Dispersion führt, also ein zumindest im wesentlichen gleichmäßiges Konzentrationsprofil des gelösten Reagenzes oder der Reaktionsprodukte in dem in den Untersuchungsbereich weiter geförderten Reaktionsvolumen erreichbar ist. Dementsprechend kann innerhalb einer definierten Zeit eine wesentlich genauere Untersuchung, insbesondere Bestimmung von Werten im genannten Sinne, erfolgen.By means of the proposed combination of measures, a substantially more accurate examination of a sample liquid, in particular a quantitative determination of at least one analyte in the sample footage, is made possible. By the defined stopping of the sample liquid in the reaction area is a Determined reaction volume in which a reagent in the reaction region is defined solvable or with which the reagent can react. Subsequently, the reaction volume with the dissolved reagent or reaction products from the reaction area in the study area further promoted, the lateral, at least substantially laminar flow and / or the at least substantially rectilinear flow front of the sample liquid leads to a low dispersion, so at least substantially uniform Concentration profile of the dissolved reagent or reaction products can be reached in the further funded in the study area reaction volume. Accordingly, within a defined time, a much more precise investigation, in particular determination of values in the aforementioned sense, can take place.

Zur Untersuchung bzw. Bestimmung können beispielsweise von dem gelösten Reagenz und einem zu bestimmenden Analyten gebildete Komplexe oder Verbindungen im Untersuchungsbereich mittels einer immobilisierten Nachweischemikalie gebunden und anschließend - beispielsweise optisch - nach-geweisen bzw. gemessen werden. Daraus läßt sich dann beispielsweise die Konzentration des Analyten in der Probenflüssigkeit bestimmen.For the purpose of investigation or determination, for example, complexes or compounds formed by the dissolved reagent and an analyte to be determined can be bound in the examination area by means of an immobilized detection chemical and subsequently - for example optically - confirmed or measured. From this, it is then possible to determine, for example, the concentration of the analyte in the sample liquid.

Ein weiterer, auch unabhängig realisierbarer Aspekt der vorliegenden Erfindung liegt darin, den Reaktionsbereich, den Untersuchungsbereich und/oder den Kanal mit einem zumindest im wesentlichen konstanten Querschnitt zu versehen und/oder die Höhe des Kanals mindestens um den Faktor 10 kleiner als die Breite des Kanals auszubilden und/oder den Reaktionsbereich und/oder den Untersuchungsbereich maximal so lang wie breit oder kürzer auszubilden.A further, independently realizable aspect of the present invention is to provide the reaction area, the examination area and / or the channel with an at least substantially constant cross-section and / or the height of the channel at least by a factor of 10 smaller than the width of the channel form and / or the reaction area and / or the examination area maximum as long as wide or shorter form.

Die vorgenannten Maßnahmen sind einer quantitativen Untersuchung bzw. biochemischen Untersuchungen zuträglich. Insbesondere wird ein definiertes Untersuchen des definierten Reaktionsvolumens ermöglicht bzw. eine unerwünschte Dispersion des Reaktionsvolumens vermieden. Weitere Vorteile sind kurze Untersuchungszeiten, schnelle Reaktionen, kurze Diffusionswege und/oder kurze Strömungswege.The above measures are conducive to quantitative analysis or biochemical investigations. In particular, a defined examination of the defined reaction volume is made possible or an undesirable dispersion of the reaction volume is avoided. Further advantages include short examination times, fast reactions, short diffusion paths and / or short flow paths.

Weitere Vorteile, Merkmale, Eigenschaften und Aspekte der vorliegenden Erfmdung ergeben sich aus den Ansprüchen und der folgenden Beschreibung bevorzugter Ausführungsformen anhand der Zeichnung. Es zeigt:

Fig. 1
einen schematischen Längsschnitt eines Teils einer vorschlagsgemäßen, teilweise mit Probenflüssigkeit gefüllten Vorrichtung gemäß einer ersten Ausführungsform;
Fig. 2
eine schematische Draufsicht eines Trägers der ungefüllten Vorrichtung gemäß Fig.1;
Fig. 3
eine zu Fig. 2 korrespondierende Draufsicht eines Trägers einer ungefüllten Vorrichtung gemäß einer zweiten Ausführungsform;
Fig. 4
einen schematischen Querschnitt der Vorrichtung gemäß der ersten Ausführungsform entlang Linie IV-IV gemäß Fig. 1;
Fig. 5
einen Fig. 4 entsprechenden, schematischen Querschnitt der Vorrichtung gemäß einer dritten Ausführungsform; und
Fig. 6
einen Fig. 3 entsprechenden, schematischen Querschnitt der Vorrichtung gemäß einer vierten Ausführungsform.
Further advantages, features, characteristics and aspects of the present invention will become apparent from the claims and the following description of preferred embodiments with reference to the drawing. It shows:
Fig. 1
a schematic longitudinal section of a portion of a proposed, partially filled with sample liquid device according to a first embodiment;
Fig. 2
a schematic plan view of a carrier of the unfilled device according to Fig.1 ;
Fig. 3
one too Fig. 2 corresponding plan view of a carrier of an unfilled device according to a second embodiment;
Fig. 4
a schematic cross section of the device according to the first embodiment along line IV-IV according to Fig. 1 ;
Fig. 5
one Fig. 4 corresponding, schematic cross section of the device according to a third embodiment; and
Fig. 6
one Fig. 3 corresponding, schematic cross section of the device according to a fourth embodiment.

In den Figuren werden für gleiche oder ähnliche Teile dieselben Bezugszeichen verwendet, wobei entsprechende oder vergleichbare Eigenschaften und Vorteile erreicht werden, auch wenn eine wiederholte Beschreibung weggelassen ist.In the figures, the same reference numerals are used for the same or similar parts, and corresponding or comparable properties and advantages are achieved, even if a repeated description is omitted.

Fig. 1 zeigt in einem schematischen Schnitt einen Teil einer ersten Ausführungsform einer vorschlagsgemäßen Vorrichtung 1 zur Untersuchung einer Probenflüssigkeit 2, insbesondere Blutplasma o. dgl. Fig. 1 shows in a schematic section a part of a first embodiment of a proposed device 1 for examining a sample liquid 2, in particular blood plasma o. The like.

Die Vorrichtung 1 weist einen die Pmbenflüssigkeit 2 durch Kapillarkräfte aufnehmenden und fördernden Kanal 3 auf. Der Kanal 3 wird vorzugsweise von nur zwei gegenüberliegenden, insbesondere im wesentlichen ebenen Flächen bzw. Flachseiten 4 und 5 begrenzt bzw. gebildet.The device 1 has a channel 3 receiving the pump fluid 2 by capillary forces. The channel 3 is preferably bounded or formed by only two opposite, in particular substantially flat surfaces or flat sides 4 and 5.

Die Vorrichtung 1 weist einen vorzugsweise plattenförmigen Träger 6 und eine zugeordnete Abdeckung 7 auf, zwischen denen der Kanal 3 gebildet ist. Beim Darstellungsbeispiel ist lediglich der Träger 6 zur Bildung der erforderlichen Mikrostrukturen ausgenommen und die Abdeckung 7 eben, vorzugsweise zumindest im wesentlichen ausnehmungsfrei, ausgebildet. Jedoch kann dies auch umgekehrt sein. Bedarfsweise können aber auch sowohl der Träger 6 als auch die Abdeckung 7 ausgenommen und/oder mit Vorsprüngen zur Bildung der gewünschten Strukturen und ggf. zur Aufnahme von nicht dargestellten Chemikalien, Reagenzien, Untersuchungseinrichtungen o. dgl. ausgebildet sein. Insbesondere handelt es sich bei der Vorrichtung also um einen sogenannten Mikrochip (Plattform mit Mikrostruktur).The device 1 has a preferably plate-shaped support 6 and an associated cover 7, between which the channel 3 is formed. In the illustrated embodiment, only the support 6 is excluded for the formation of the required microstructures and the cover 7 flat, preferably at least substantially ausnehmungsfrei formed. However, this can be the other way around. If necessary, however, both the carrier 6 and the cover 7 can be excluded and / or formed with projections for forming the desired structures and, if necessary, for holding chemicals, reagents, examination devices or the like, not shown. In particular, the device is thus a so-called microchip (platform with microstructure).

Fig. 2 zeigt in einer schematischen Draufsicht den Träger 6 der Vorrichtung 1, jedoch ohne Abdeckung 7 und ohne Probenflüssigkeit 2. Der Kanal 3 weist vorzugsweise unmittelbar nacheinander einen Einlaß, beim Darstellungsbeispiel insbesondere mit einem Graben 8, einen Reaktionsbereich 9, einen Untersuchungsbereich 10 und/oder einen Sammelbereich 11 auf. Fig. 2 shows in a schematic plan view of the carrier 6 of the device 1, but without cover 7 and without sample liquid 2. The channel 3 preferably has one inlet immediately following, in the illustrated example in particular with a trench 8, a reaction area 9, an examination area 10 and / or a Collection area 11 on.

Bedarfsweise kann eine nicht dargestellte Dosierung der Probenflüssigkeit 2 in den Kanal 3 oder im Kanal 3, insbesondere vor dem Einlaß, erfolgen, wie beispielsweise in der EP 1440 732 Al beschrieben.If necessary, a dosage of the sample liquid 2, not shown, in the channel 3 or in the channel 3, in particular in front of the inlet, take place, such as in the EP 1440 732 Al described.

Die Vorrichtung 1 weist vorzugsweise nur einen einzigen Kanal 3 auf. Hierbei ist der Kanal 3 im Sinne einer Einzelkapillare zu verstehen. Bedarfsweise kann der Kanal 3 jedoch in unterschiedliche Richtungen oder zu unterschiedlichen Bereichen führen oder sich verzweigen. Beim Darstellungsbeispiel strömt die Probenflüssigkeit 2 vorzugsweise ausschließlich durch Kapillarkräfte in den bzw. im Kanal 3 in Strömungsrichtung S, wie in Fig. 1 angedeutet. Jedoch kann die Probenflüssigkeit 2 zusätzlich oder alternativ auch beispielsweise durch Druck durch den Kanal 3 gefördert werden.The device 1 preferably has only a single channel 3. Here, the channel 3 is to be understood in terms of a single capillary. If necessary, however, the channel 3 can lead in different directions or to different areas or branch. In the illustrated example, the sample liquid 2 preferably flows exclusively through capillary forces into or into the channel 3 in the flow direction S, as in FIG Fig. 1 indicated. However, the sample liquid 2 may additionally or alternatively also be conveyed, for example, by pressure through the channel 3.

Der Kanal 3 weist vorzugsweise einen im wesentlichen rechteckigen und/oder flachen Querschnitt quer zur Strömungsrichtung S der Probenflüssigkeit 2 auf.The channel 3 preferably has a substantially rectangular and / or flat cross section transverse to the flow direction S of the sample liquid 2.

Die in Fig. 1 bzw. 4 angedeutete Höhe H des Kanals 3 - also der Abstand der den Kanal 3 begrenzenden, vorzugsweise parallelen Flächen 4 und 5 - beträgt maximal 2000 µm, vorzugsweise höchstens 500 µm, insbesondere etwa 50 bis 200 µm. Die Breite des Kanals 3 beträgt vorzugsweise etwa 100 bis 5000 µm, insbesondere etwa 200 bis 4000 µm. Die Höhe H des Kanals 3 ist wesentlich geringer, insbesondere mindestens um den Faktor 10 oder 100, als die Breite des Kanals 3. Das Aufuahmevolumen des Kanals 3 beträgt vorzugsweise weniger als 1 ml, insbesondere weniger als 100 µl, besonders bevorzugt maximal 10 µl.In the Fig. 1 or 4 indicated height H of the channel 3 - ie the distance of the channel 3 bounding, preferably parallel surfaces 4 and 5 - is at most 2000 microns, preferably at most 500 .mu.m, in particular about 50 to 200 microns. The width of the channel 3 is preferably about 100 to 5000 microns, in particular about 200 to 4000 microns. The height H of the channel 3 is substantially smaller, in particular at least by a factor of 10 or 100, than the width of the channel 3. The absorption volume of the channel 3 is preferably less than 1 ml, in particular less than 100 μl, particularly preferably not more than 10 μl.

Die Vorrichtung 1 bildet also ein mikrofluidisches System. Insbesondere dient die Vorrichtung 1 der mikrofluidischen Diagnostik für medizinische oder nicht-medizinische Zwecke bzw. sonstigen Untersuchungen.The device 1 thus forms a microfluidic system. In particular, the device 1 is used for microfluidic diagnostics for medical or non-medical purposes or other examinations.

Der Kanal 3 und dessen Haupterstreckungsebene verlaufen in Gebrauchslage vorzugsweise zumindest im wesentlichen horizontal. Je nach Verwendungszweck oder konstruktiver Lösung ist jedoch auch eine andere Ausrichtung möglich, zurnal die Aufnahme bzw. das Füllen des Kanals 3 mit Probenflüssigkeit 2 vorzugsweise zumindest primär nur durch Kapillarkräfte bestimmt bzw. bewirkt wird.The channel 3 and its main extension plane preferably extend at least substantially horizontally in the position of use. Depending on the intended use or constructive solution, however, a different orientation is possible, especially since the reception or filling of the channel 3 with sample liquid 2 is preferably determined or effected at least primarily by capillary forces.

Der Reaktionsbereich 9 mit einem von der Probenflüssigkeit 2 lösbaren und/oder damit reagierenden Reagenz und der sich vorzugsweise unmittelbar anschließende Untersuchungsbereich 10 sind im Kanal 3, vorzugsweise nacheinander auf derselben Flachseite 4 des Kanals 3, gebildet.The reaction region 9 with a reagent which can be dissolved and / or reacted with the sample liquid 2 and the preferably immediately adjoining examination region 10 are formed in the channel 3, preferably successively on the same flat side 4 of the channel 3.

Der Reaktionsbereich 9 weist ein von der Probenflüssigkeit 2 vorzugsweise lösbares Reagenz für einen zu einem zu bestimmenden Analyten in der Probenflüssigkeit 2 auf. Insbesondere handelt es sich bei dem Reagenz um gegen den zu messenden Analyten gerichtete Antikörper, die an Indikatoren (Farbstoffe, Farbstoffpartikel, z.B. kolloidales Gold) gebunden sind. Im Reaktionsbereich 9 wird beim Füllen mit der Probenflüssigkeit 2 das Reagenz gelöst. Der Analyt, wenn er in der Probenflüssigkeit 2 enthalten ist, reagiert dann mit dem am Farbstoff gebundenen Antikörper und bildet insbesondere eine Verbindung oder einen Komplex.The reaction region 9 has a reagent, which is preferably detachable from the sample liquid 2, for an analyte to be determined in the sample liquid 2. In particular, the reagent is antibodies directed against the analyte of interest and bound to indicators (dyes, colorant particles, e.g., colloidal gold). In the reaction area 9, the reagent is dissolved when filled with the sample liquid 2. The analyte, when contained in the sample fluid 2, then reacts with the antibody bound to the dye, and in particular forms a compound or complex.

Alternativ oder zusätzlich reagiert das Reagenz mit dem Analyten und bildet insbesondere ein Reaktionsprodukt, auch wenn das Reagenz ggf. nicht gelöst wird. Die nachfolgenden Ausführungen hinsichtlich des (gelösten) Reagenzes gelten daher für das Reaktionsprodukt entsprechend.Alternatively or additionally, the reagent reacts with the analyte and in particular forms a reaction product, even if the reagent is not dissolved if necessary. The following statements with regard to the (dissolved) reagent therefore apply correspondingly to the reaction product.

Der Untersuchungsbereich 10 ist beim Darstellungsbeispiel mit einer vorzugsweise immobilisierten Nachweischemikalie versehen, die insbesondere Verbindungen oder Komplexe aus Analyt und Reagenz bzw. das Reaktionsprodukt bindet. Ungebundenes Reagenz und sonstige Bestandteile strömen mit der Probenflüssigkeit 2 anschließend weiter in den Sammelbereich 11, wo sie aufgenommen werden und dadurch ein Rückfluß verhindert wird. Im Untersuchungsbereich 10 sind dann beispielsweise optisch das gebundene Reagenz und daraus das Vorhandensein und insbesondere die Konzentration des Analyten in der Probenflüssigkeit 2 bestimmbar. Es wird also eine insbesondere quantitative Untersuchung der Probenflüssigkeit 2 ermöglicht.In the case of the illustration, the examination region 10 is provided with a preferably immobilized detection chemical, which in particular binds compounds or complexes of analyte and reagent or the reaction product. Unbound reagent and other ingredients then flow with the sample fluid 2 further into the collection area 11, where they are taken up, thereby preventing backflow. In the examination region 10, for example, the bound reagent can be determined optically and from this the presence and in particular the concentration of the analyte in the sample liquid 2 can be determined. Thus, a particularly quantitative examination of the sample liquid 2 is made possible.

Die Vorrichtung 1 weist vorschlagsgemäß eine Einrichtung 12 zum temporären Anhalten der Probenfüßigkeit 2 im Reaktionsbereich 9 zum Lösen und/oder Reagieren des Reagenzes und/oder im Untersuchungsbereich 10 aufAccording to the proposal, the device 1 has a device 12 for temporary stopping of the sample foot 2 in the reaction area 9 for dissolving and / or reacting the reagent and / or in the examination area 10

Die Einrichtung 12 ist vorzugsweise derart ausgebildet, daß das temporäre Anhalten durch die Probenflüssigkeit 2 selbst, wie beispielsweise in der EP 1 441 131 Al beschrieben, oder durch eine nicht dargestellte Steuerflüssigkeit oder durch eine selektive Entlüftung, wie beispielsweise in der EP 1419 818 Al beschrieben, festlegbar bzw, aufhebbar ist. Vorzugsweise hält die Einrichtung 12 die Probenflüssigkeit 2 für eine vorbestimmte Zeitdauer, ggf. erst nach dem vollständigen Füllen des Reaktionsbereichs 9, und/oder bis zum vollständigen Füllen des Reaktionsbereichs 9 an.The device 12 is preferably designed such that the temporary stop by the sample liquid 2 itself, such as in the EP 1 441 131 Al described, or by a control fluid, not shown, or by a selective vent, such as in the EP 1419 818 Al described, fixed or can be canceled. Preferably, the device 12 holds the sample liquid 2 for a predetermined period of time, possibly only after the complete filling of the reaction zone 9, and / or until the reaction zone 9 has been completely filled.

Beim Darstellungsbeispiel weist die Einrichtung 12 insbesondere einen Steuerkanal 13 auf, der Probenflüssigkeit 2 nach bzw. in einer definierte Zeit einem zwischen dem Reaktionsbereich 9 und dem Untersuchungsbereich 10 angeordneten Flüssigkeitstopp 14 zuführt, so daß dann die Probenflüssigkeit 2 bzw. daß im Reaktionsbereich 9 befindliche Reaktionsvolumen an Probenflüssigkeit 2 den Flüssigkeitsstopp 14 überwinden und in den Untersuchungsbereich 10 weiterströmen kann.In the illustrated example, the device 12 has, in particular, a control channel 13 which supplies sample liquid 2 to a liquid stop 14 arranged between the reaction zone 9 and the examination zone 10 within a defined time, so that then the sample liquid 2 or reaction volume located in the reaction zone 9 on sample liquid 2 overcome the liquid stop 14 and can flow into the examination area 10.

Bedarfsweise weist die Vorrichtung 1 eine weitere Einrichtung 12 zum temporären Anhalten der Probenflüssigkeit 2 - insbesondere des aus dem Reaktionsbereich 9 in den Untersuchungsbereich 10 zuvor geströmten Reaktionsvolumens an Probenflüssigkeit 2, das gelöstes Reagenz oder das Reaktionsprodukt enthält - im Untersuchungsbereich 10 auf, um eine möglichst genaue bzw. quantitative Bestimmung zu ermöglichen, insbesondere um eine zumindest im wesentlichen vollständige Reaktion bzw. Bindung der Verbindungen oder Komplexe aus Reagenz und Analyt oder des Reaktionsprodukts an der Nachweischemikalie im Untersuchungsbereich 10 zu ermöglichen.If necessary, the device 1 has a further device 12 for temporary stopping of the sample liquid 2 - in particular the reaction volume of sample liquid 2 which has previously flowed from the reaction region 9 into the examination region 10 and contains dissolved reagent or the reaction product - in the examination region 10 in order to obtain the most accurate possible or to permit quantitative determination, in particular in order to enable an at least substantially complete reaction or binding of the compounds or complexes of reagent and analyte or of the reaction product to the detection chemical in the examination area 10.

Die weitere Einrichtung 12 ist insbesondere entsprechend der vorgenannten Einrichtung 12 ausgebildet. Entsprechend ist wiederum ein Steuerkanal 13 vorgesehen, der Probenflüssigkeit 2 nach bzw. in einer definierten Zeit einem zwischen dem Untersuchungsbereich 10 und dem nachgeordneten Sammelbereich 11 angeordneten Flüssigkeitsstopp 14 zuführt, so daß dann die Probenflüssigkeit 2 bzw. das im Untersuchungsbereich 10 befindliche Reaktionsvolumen an Probenflüssigkeit 2 den Flüssigkeitsstopp 14 überwinden und in den Sammelbereich 11 weiterströmen und die nachströmende Probenflüssigkeit 2 dann ein Auswaschen von nicht gebundenem Reagenz bzw. Reaktionsprodukt in den Untersuchungsbereich 10 bewirken kann.The further device 12 is designed in particular according to the aforementioned device 12. Accordingly, in turn, a control channel 13 is provided, the sample liquid 2 after or in a defined time to a arranged between the examination area 10 and the downstream collection area 11 liquid stop 14 supplies, so that then the sample liquid 2 or located in the examination area 10 reaction volume of sample liquid. 2 overcome the liquid stop 14 and continue to flow into the collection area 11 and the inflowing sample liquid. 2 then a washing out of unbound reagent or reaction product in the examination area 10 can cause.

Beim Darstellungsbeispiel ist der Flüssigkeitsstopp 14 insbesondre durch eine nut- oder grabenartige Vertiefung quer zur Strömungsrichtung S gebildet. Jedoch sind auch andere konstruktive Lösungen möglich. Insbesondere sind aus der US 5,458,852 weitere konstruktive Lösungen zur Realisierung der Einrichtung(en) 16 bekannt, die alternativ oder zusätzlich eingesetzt werden können.In the illustrated example, the liquid stop 14 is formed in particular by a groove-like or trench-like depression transverse to the flow direction S. However, other constructive solutions are possible. In particular, are from the US 5,458,852 Further constructive solutions for the realization of the device (s) 16 known, which can be used alternatively or additionally.

Fig. 3 zeigt in einer zu Fig. 2 korrespondierenden Draufsicht auf den ungefüllten Träger 6 einer Vorrichtung 1 gemäß einer zweiten Ausführungsform. Die Einrichtungen 12 weisen hier jeweils anstatt eines vorzugsweise nut- oder grabenartigen Flüssigkeitsstopps 14 mindestens eine vorzugsweise stegartige Barriere 15, insbesondere zwei oder mehr hintereinander angeordnete Barrieren 15, auf. So kann bedarfsweise ein entsprechendes temporäres Anhalten der Probenflüssigkeit 2 erreicht werden. Fig. 3 shows in one too Fig. 2 corresponding plan view of the unfilled carrier 6 of a device 1 according to a second embodiment. The devices 12 each have here, instead of a preferably groove-like or trench-like liquid stop 14, at least one preferably web-like barrier 15, in particular two or more barriers 15 arranged one behind the other. Thus, if necessary, a corresponding temporary stopping of the sample liquid 2 can be achieved.

Bei der zweiten Ausführungsform weist der Kanal 3 im Gegensatz zu der ersten Ausführungsform keinen im wesentlichen konstanten Querschnitt auf. Vielmehr ist der Querschnitt des Kanals 3 am Übergang vom Reaktionsbereich 9 zum Untersuchungsbereich 10 und/oder beim Übergang vom Untersuchungsbereich 10 in den Sammelbereich 11 verringert. Diese Querschnittsverringerung wird vorzugsweise durch gleichmäßige Verjüngung des Flüssigkeitsstroms und anschließende Aufspreizung des Flüssigkeitsstroms erreicht. Die Einrichtung 12 bzw. Barriere 15 ist dann vorzugsweise im Bereich des verringerten Querschnitts angeordnet.In the second embodiment, the channel 3 does not have a substantially constant cross-section, in contrast to the first embodiment. Instead, the cross section of the channel 3 at the transition from the reaction region 9 to the examination region 10 and / or during the transition from the examination region 10 into the collection region 11 is reduced. This reduction in cross-section is preferably achieved by uniform tapering of the liquid stream and subsequent spreading of the liquid stream. The device 12 or barrier 15 is then preferably arranged in the region of the reduced cross section.

Die genannte Querschnittsverringerung führt bereits zu einer Verringerung des Volumenstroms durch den Kanal 3, so daß ggf. kein vollständiges temporäres Anhalten erforderlich ist. Insbesondere kann eine durch die Barrieren 15 verursachte Verzögerung der Strömung bzw. Reduktion des Volumenstroms bedarfsweise genügen.Said cross-sectional reduction already leads to a reduction in the volume flow through the channel 3, so that, if necessary, no complete temporary stop is required. In particular, a delay caused by the barriers 15 of the flow or reduction of the volume flow may be sufficient.

Die Vorrichtung 1 gemäß der ersten oder zweiten Ausführungsform weist weiter ein Mittel 16 zum Verhindern des seitlichen Vorschießens von Probenflüssigkeit 2 in Strömungsrichtung S und/oder zur Erzeugung einer in Fig. 1 angedeuteten, bezüglich der Draufsicht gemäß Fig. 2 wenig gekrümmten oder geradlinigen Strömungsfront F der Probenflüssigkeit 2 bzw. zur Erzeugung einer homogenen bzw. laminaren Strömung auf.The device 1 according to the first or second embodiment further comprises a means 16 for preventing the lateral advancement of sample liquid 2 in the flow direction S and / or for generating an in Fig. 1 indicated, according to the plan view Fig. 2 little curved or rectilinear flow front F of the sample liquid 2 or to produce a homogeneous or laminar flow.

Beim Darstellungsbeispiel ist das Mittel 15 dadurch gebildet, daß der Kanal 3 zumindest längsseitig offen ausgebildet ist. Seitlich an den Kanal 3 schließt sich eine Ausnehmung 17 an, die insbesondere nut- bzw. grabenartig ausgebildet ist. So wird ein seitlicher Flüssigkeitsstopp für die Probenflüssigkeit 2 - also ein durch Kapillarkräfte nicht überwindbares Strömungshindemis - gebildet und die Probenflüssigkeit 2 seitenwandfrei entlang der offenen Längsseiten im Kanal 3 geführt.In the illustrated example, the means 15 is formed in that the channel 3 is formed open at least on the long side. The side of the channel 3 is followed by a recess 17, which is formed in particular nut- or trench-like. Thus, a lateral liquid stop for the sample liquid 2 - that is, a flow impediment that can not be overcome by capillary forces - is formed and the sample liquid 2 is guided along the open longitudinal sides in the channel 3 without sidewalls.

Die Ausnehmung 17 schließt sich vorzugsweise scharfkantig an den Kanal 3 an, wie in Fig. 1, 3 und 4 angedeutet. Beim Darstellungsbeispiel ist die Ausnehmung 17 nur im Träger 6 gebildet, erstreckt sich bei der Darstellung gemäß Fig, 1, 3 und 4 also im wesentlichen nur nach unten bezüglich einer seitlichen Projektion des Kanals 3. Die Ausnehmung 17 kann sich jedoch wahlweise auch nach oben oder auf beide Seiten der seitlichen Projektion des Kanals 3, also insbesondere nach oben und nach unten erstrecken.The recess 17 preferably connects sharp-edged to the channel 3, as in Fig. 1 . 3 and 4 indicated. In the illustrated example, the recess 17 is formed only in the carrier 6, extends in the illustration according to Fig, 1 . 3 and 4 Thus, essentially only downwardly with respect to a lateral projection of the channel 3. The recess 17 may, however, optionally also upwards or on both sides of the lateral projection of the channel 3, so in particular extend upwards and downwards.

Die im Querschnitt vorzugsweise rechteckige Ausnehmung 17 führt zu einer derartigen, insbesondere stufigen bzw. plötzlichen Querschnittsvergrößerung, daß sich die Kapillarkräfte derartig verringern, daß der genannte Flüssigkeitsstopp für die Probenflüssigkeit 2 im Übergang von Kanal 3 zur Ausnehmung 17 hin gebildet wird. Insbesondere ist die Höhe der Ausnehmung 17 mindestens doppelt so groß wie die Höhe H des Kanals 3.The preferably rectangular cross-section recess 17 leads to such, in particular stepped or sudden cross-sectional enlargement that reduce the capillary forces such that said liquid stop for the sample liquid 2 is formed in the transition from channel 3 to the recess 17 out. In particular, the height of the recess 17 is at least twice as large as the height H of the channel 3.

Die Ausnehmung 17 erstreckt sich beim Darstellungsbeispiel entlang der offenen Seite des Kanals 3, insbesondere ist sie umlaufend um den allseitig offenen Kanal 3 ausgebildet.The recess 17 extends in the representation example along the open side of the channel 3, in particular, it is formed circumferentially around the channel 3 open on all sides.

Eine entsprechende seitenwandlose Führung der Probenflüssigkeit 2 im Kanal 3 ist auch bei der in Fig. 5 dargestellten, dritten Ausführungsform der Vorrichtung 1 durch die seitliche Ausnehmung 17 möglich. Die Probenflüssigkeit 2 wird hier jedoch nur auf einer Boden- bzw. Flachseite 4 geführt. Die Probenflüssxgkeit 2 steht also nicht in Kontakt mit einer gegenüberliegenden Flachseite 5 wie bei der ersten Ausführungsform. Statt dessen ist beim Darstellungsbeispiel die Abdeckung 7 entsprechend ausgenommen oder die Fläche 4 entsprechend tief im Träger 6 angeordnet, um einen genügenden Abstand zur dann ggf. ebenen Abdeckung 7 einhalten zu können. Die Dicke des von der Probenflüssigkeit 2 auf der Fläche 4 gebildete Flüssigkeitsfilms hängt insbesondere vom Benetzungsverhalten und von der zugeführten, dann insbesondere zudosierten Menge an Probenflüssigkeit 2 ab. Vorzugsweise gelten für den Flüssigkeitsfilm dann die entsprechenden Dimensionen, wie bei der ersten Ausführungsform für den Kanal 3 erläutert.A corresponding side wallless leadership of the sample liquid 2 in the channel 3 is also in the in Fig. 5 illustrated, third embodiment of the device 1 through the lateral recess 17 is possible. However, the sample liquid 2 is guided here only on a bottom or flat side 4. The sample liquid 2 is thus not in contact with an opposite flat side 5 as in the first embodiment. Instead, in the illustrated embodiment, the cover 7 is excluded accordingly or the surface 4 is arranged correspondingly deep in the carrier 6 in order to maintain a sufficient distance to the then possibly flat cover 7 can. The thickness of the liquid film formed by the sample liquid 2 on the surface 4 depends in particular on the wetting behavior and on the quantity of sample liquid 2 fed in, in particular metered in. The corresponding dimensions then apply to the liquid film, as explained for the channel 3 in the first embodiment.

Fig. 6 zeigt in einem schematischen Schnitt eine vierte Ausführungsform der vorschlagsgemäßen Vorrichtung 1, wobei ein Seitenbereich des Kanals 3 zur Verdeutlichung vergrößert dargestellt ist. Fig. 6 shows in a schematic section a fourth embodiment of the proposed device 1, wherein a side portion of the channel 3 is shown enlarged for clarity.

Das Mittel 16 zum Verhindern des seitlichen Vorschießens von Probenflüssigkeit 2, zur Erzeugung einer wenig gekrümmten oder geradlinigen Strömungsfront F und/oder zur Erzeugung einer homogenen bzw. laminaren Strömung kann alternativ oder zusätzlich auch eine den Kanal 3 längsseitig oder allseitig begrenzende Seitenwandung 18 aufweisen, wobei durch Ausbildung von entsprechenden Leitelementen oder Verzögerungsstrukturen, insbesondere Vorsprüngen oder Erhebungen 19 o.dgl., die Strömungs- bzw. Füllgeschwindigkeit entlang der Seitenwandung 18 in Strömungsrichtung S die Füllgeschwindigkeit verringert, insbesondere so daß die Füllgeschwindigkeit der Probenflüssigkeit 2 am Rand diejenige im mittleren Bereich des Kanals 3 nicht überschreitet, sondern dieser zumindest im wesentlichen entspricht. Alternativ oder zusätzlich zu den Leitelementen bzw. Verzögerungsstrukturen kann die Benetzung der Seitenwand 18 auch derart modifiziert, insbesondere verringert, sein, daß das unerwünschte Vorschießen von Probenflüssigkeit 2 entlang der Seitenwand 18 vermieden wird.The means 16 for preventing the lateral advancement of sample liquid 2, for producing a slightly curved or rectilinear flow front F and / or for generating a homogeneous or laminar flow may alternatively or additionally also have a side wall 18 which delimits the channel 3 on the long side or on all sides by forming corresponding guide elements or delay structures, in particular projections or elevations 19 or the like., The flow or filling speed along the side wall 18 in the flow direction S reduces the filling speed, in particular so that the filling rate of the sample liquid 2 at the edge that in the central region of Channel 3 does not exceed, but this at least substantially corresponds. As an alternative or in addition to the guide elements or delay structures, the wetting of the side wall 18 can also be modified, in particular reduced, in such a way that the undesirable advancing of sample liquid 2 along the side wall 18 is avoided.

Ergänzend wird hinsichtlich möglicher Ausgestaltungen, um ein Vorschießen der Probenflüssigkeit 2 längsseitig zu vermeiden, auf die diesbezüglich in der EP 1201304 A2 beschriebenen Möglichkeiten verwiesen.In addition, with regard to possible embodiments, in order to avoid a pre-shooting of the sample liquid 2 along the side, in this regard in the EP 1201304 A2 referenced options.

Bei den Darstellungsbeispielen weist der Kanal 3 mindestens ein Leitelement zur Beeinflussung, insbesondere Vergleichmäßigung, des Füllens mit der Probenflüssigkeit 2 auf. Insbesondere weist der Kanal 3 vorzugsweise regelmäßig verteilte Erhebungen 19 als Leitelemente auf der Flachseite 4 oder ggf. beiden Flachseiten 4, 5 auf, wie in Fig. 1, 2 und 4 bis 6 dargestellt. Diese sind insbesondere in Reihen quer, vorzugsweise senkrecht, oder längs zur Strömungsrichtung S, insbesondere abwechselnd quer versetzt, angeordnet. So kann erreicht werden, daß die Probenflüssigkeit 2 den Kanal 3 reihenweise - also Reihe für Reihe - füllt und dadurch mit einer im wesentlichen gradlinigen Flüssigkeitsfront F in Strömungsrichtung S fortschreitet. Das Mittel 16 umfaßt bedarfsweise auch die genannten Leitelemente.In the illustration examples, the channel 3 has at least one guide element for influencing, in particular equalizing, the filling with the sample liquid 2. In particular, the channel 3 preferably has regularly distributed elevations 19 as guide elements on the flat side 4 or possibly both flat sides 4, 5, as in FIG Fig. 1, 2 and 4 to 6 shown. These are in particular arranged in rows transversely, preferably vertically, or longitudinally to the flow direction S, in particular alternately transversely offset. Thus, it can be achieved that the sample liquid 2 fills the channel 3 in rows, ie row by row, and thereby advances with a substantially straight-line liquid front F in the flow direction S. The means 16 also includes, if necessary, said guide elements.

Bedarfsweise kann die Flächendichte, der Abstand und/oder die Größe der Erhebungen 19 variieren, insbesondere in Abhängigkeit von der jeweiligen Entfernung zum Einlaß, um einen gewünschten Verlauf der Kapillarkräfte bzw. ggf. eine Kompensation von Strömungswiderständen zu erreichen.If necessary, the area density, the distance and / or the size of the elevations 19 vary, in particular as a function of the respective distance to the inlet, in order to achieve a desired course of the capillary forces or possibly a compensation of flow resistances.

Die Erhebungen 19 sind vorzugsweise steg-, höcker- oder säulenartig, insbesondere mit runder oder polygonaler Grundfläche, ausgebildet. Alternativ oder zusätzlich können aber auch Vertiefungen, wie der Graben 8, oder die Barrieren 15 oder sonstige Leitelemente vorgesehen sein, die quer oder längs zur Strömungsnchtung S des Kanals 3 verlaufen.The elevations 19 are preferably web-like, hump-shaped or column-like, in particular with a round or polygonal base surface. Alternatively or additionally, however, it is also possible to provide depressions, such as the trench 8, or the barriers 15 or other guide elements, which extend transversely or longitudinally to the flow direction S of the channel 3.

Der vorzugsweise vorgesehene nutartige, im Querschnitt insbesondere rechteckige oder halbrunde Graben 8 weist eine wesentlich geringere Tiefe als der Flüssigkeitsstopp 14 und die Ausnehmung 17 auf und bildet daher einen nur temporären Flüssigkeitsstopp zur Vergleichmäßigung der Flüssigkeitsfront F. So kann erreicht werden, daß die Probenflüssiglcezt 2 erst nach Füllen des Kanals 3 über den gesamten Querschnitt den Graben 8 und anschließend den nachfolgenden Kanalbereich füllt.The preferably provided groove-like, in cross-section in particular rectangular or semicircular trench 8 has a much smaller depth than the liquid stop 14 and the recess 17 and therefore forms only a temporary liquid stop to equalize the liquid front F. Thus, it can be achieved that the Probenflüssiglcezt 2 only after filling the channel 3 over the entire cross section fills the trench 8 and then the subsequent channel region.

Hervorzuheben ist, daß durch die Kombination der seitenwandlosen Führung der Probenflüssigkeit 2 und der Leitelemente ein hochgradig gleichmäßiges Füllen des Kanals 3 insbesondere durch Kapillarkräfte mit zumindest im wesentlichen gradliniger bzw. senkrecht zur Strömungsrichtung S verlaufender Flüssigkeitsfront F erreichbar ist.It should be emphasized that a highly uniform filling of the channel 3 in particular by capillary forces with at least substantially straight-line or perpendicular to the flow direction S extending liquid front F can be achieved by the combination of side wallless guidance of the sample liquid 2 and the guide elements.

Alternativ kann der Kanal 3 bereichsweise oder insgesamt auch zumindest im wesentlichen glatt bzw. eben, also insbesondere ohne Leitelemente, ausgebildet sein, wie in Fig. 3 angedeutet.Alternatively, the channel 3 can be formed partially or even at least substantially smoothly or even, that is to say in particular without guide elements, as in FIG Fig. 3 indicated.

Die Strukturierung bzw. Texturierung des Reaktionsbereichs 9 und/oder des Untersuchungsbereichs 10, insbesondere durch Leitelemente, wie die Erhöhungen 19 o.dgl., erleichtert das vorzugsweise gleichmäßige Aufbringen einer Chemikalie o.dgl., die anschließend eintrocknet und dadurch beispielsweise eine Trockenchemikalie oder immobilisierte Chemikalie bildet.The structuring or texturing of the reaction area 9 and / or the examination area 10, in particular by guide elements, such as the elevations 19 or the like, facilitates the preferably uniform application of a chemical or the like, which subsequently dries and thereby, for example, a dry chemical or immobilized Chemical forms.

Ergänzend ist anzumerken, daß die Vorrichtung 1 vorzugsweise eine mit der Ausnehmung 17 in Verbindung stehende Entlüftung 20 aufweist, wie in Fig. 4 angedeutet. Dies gestattet auf sehr einfache Weise eine wirksame Entlüftung. Dies ist einem gleichmäßigen, blasenfreien Füllen des Kanals 3 mit der Probenflüssigkeit 2 zuträglich.In addition, it should be noted that the device 1 preferably has a vent 20 in communication with the recess 17, as in FIG Fig. 4 indicated. This allows in a very simple way effective ventilation. This is conducive to a uniform, bubble-free filling of the channel 3 with the sample liquid 2.

Nachfolgend wird das Zusammenwirken der vorschlagsgemäßen Maßnahmen näher erläutert, wobei insbesondere auf die erste Ausführungsform bzw. die Darstellung gemäß Fig. 1 und Fig. 2 Bezug genommen wird.The interaction of the proposed measures will be explained in more detail below, with particular reference to the first embodiment and the illustration according to Fig. 1 and Fig. 2 Reference is made.

Nach dem Einfüllen wird die Probenflüssigkeit 2 durch Kapillarkräfte im Kanal 3 in den Reaktionsbereich 9 geleitet. Hierbei strömt die Probenflüssigkeit 2 im Kanal 3 - zumindest im Reaktionsbereich 9 und Untersuchungsbereich 10 - lateral über die Flachseite 4 und vorzugsweise zumindest im wesentlichen laminar bzw. mit homogener Strömungsgeschwindigkeit bzw. wenig gekrümmter oder gradliniger Strömungsfront F. Dies wird insbesondere durch das genannte Mittel 16 - insbesondere in Kombination mit den vorzugsweise zumindest im Reaktionsbereich 9 und/oder Untersuchungsbereich 10 vorgesehenen Leitelementen - erreicht.After filling, the sample liquid 2 is conducted into the reaction zone 9 by capillary forces in the channel 3. In this case, the sample liquid 2 flows in the channel 3 - at least in the reaction region 9 and examination region 10 - laterally over the flat side 4 and preferably at least substantially laminar or with homogeneous flow velocity or slightly curved or rectilinear flow front F. This is in particular by said means 16 - In particular in combination with the preferably at least in the reaction region 9 and / or examination area 10 provided guide elements - achieved.

Im Reaktionsbereich 9 wird die Probenflüssigkeit 2 für eine vorzugsweise vorbestimmte Zeit mittels der Einrichtung 12 temporär angehalten. Im Reaküonsbereich 9 kann die Probenflüssigkeit 2 das vorzugsweise als Trockenchemikalie vorliegende Reagenz zur Bestimmung eines Analyten in der Probenflüssigkeit 2 lösen bzw. damit reagieren. Bei dem Reagenz handelt es sich beispielsweise um ein Konjugat, das aus einem an dem Analyten bindenden Antikörper und aus einem Farbstoffpartikel o.dgl. gebildet ist. Das gelöste Reagenz bzw. Konjugat bindet dann an den Analyten.In the reaction region 9, the sample liquid 2 is temporarily stopped by the device 12 for a preferably predetermined time. In the reaction area 9, the sample liquid 2 can dissolve or react with the reagent, which is preferably present as a dry chemical, for determining an analyte in the sample liquid 2. The reagent is, for example a conjugate composed of an antibody binding to the analyte and a dye particle or the like. is formed. The dissolved reagent or conjugate then binds to the analyte.

Durch das temporäre Anhalten der Probenflüssigkeit 2 ist der Reaktionsbereich 9 mit einem definierten Reaktionsvolumen an Probenfüßigkeit 2 gefüllt, so daß sich das Reagenz zumindest im wesentlichen nur in diesem Reaktionsvolumen löst bzw. nur mit diesem reagiert. Durch das temporäre Anhalten kann damit bereits eine unerwünschte Dispersion bzw, weitläufige Verteilung des Reagenzes oder eines Reaktionsprodukts des Reagenzes in der Probenflüssigkeit 2 vermieden werden.Due to the temporary stopping of the sample liquid 2, the reaction area 9 is filled with a defined reaction volume of the sample foot 2, so that the reagent dissolves at least substantially only in this reaction volume or only reacts with it. As a result of the temporary stop, an undesired dispersion or extensive distribution of the reagent or a reaction product of the reagent in the sample liquid 2 can thus already be avoided.

Des weiteren ist die Zeit für das temporäre Anhalten vorzugsweise derart gewählt, daß das Reagenz zu mindestens 90 %, insbesondere zu mindestens 95 % oder wesentlich mehr insbesondere in dem Reaktionsvolumen von Probenflüssigkeit 2 gelöst wird bzw. mit diesem reagiert. Bedarfsweise kann das Lösen oder Reagieren durch Wärme oder sonstige Maßnahmen, wie Anlegen einer Spannung o.dgl., unterstützt werden.Furthermore, the time for the temporary stop is preferably chosen such that the reagent is dissolved to at least 90%, in particular at least 95% or substantially more in particular in the reaction volume of sample liquid 2 or reacts with it. If necessary, the dissolution or reaction by heat or other measures, such as applying a voltage or the like, Can be supported.

Das Reagenz ist vorzugsweise gleichmäßig oder in einer vorbestimmten Konzentrationsverteilung auf der Flachseite 4 im Reaktionsbereich 9 aufgebracht. So kann erreicht werden, daß - ggf. unter Berücksichtigung des Füllvorgangs durch die Probenflüssigkeit 2 - eine möglichst schnelle und gleichmäßige Verteilung des gelösten Reagenzes bzw. des Reaktionsprodulcts in dem genannten Reaktionsvolumen erreicht wird.The reagent is preferably applied uniformly or in a predetermined concentration distribution on the flat side 4 in the reaction region 9. Thus, it can be achieved that - if necessary, taking into account the filling process by the sample liquid 2 - as fast and uniform distribution of the dissolved reagent or the Reaktionsprodulcts is achieved in said reaction volume.

Der möglichst gleichmäßigen Verteilung in dem Reaktionsvolumen ist auch zuträglich, daß das Reagenz auf der Flachseite 4 des Kanals 3 angeordnet ist und daß aufgrund der verhältnismäßig geringen Kanalhöhe H dementsprechend eine schnelle Diffusion und dadurch gleichmäßige Verteilung des gelösten Reagenzes bzw. des Reaktionsprodukts im Reaktionsvolumen erreichbar sind.The most even distribution in the reaction volume is also beneficial that the reagent on the flat side 4 of the channel 3 is arranged and that due to the relatively low channel height H accordingly a fast diffusion and thus uniform distribution of the dissolved reagent or the reaction product in the reaction volume can be achieved ,

Nach dem definierten Lösen bzw. Reagieren gibt die Einrichtung 12 die Probenflüssigkeit 2 frei, so daß die Probenflüssigkeit 2 - insbesondere das definierte Reaktionsvolumen - von dem Reaktionsbereich 9 in den Untersuchungsbereich 10 weiterströmen kann. Aufgrund der genannten Maßnahmen und insbesondere des Mittels 16 kann dabei wiederum eine besonders geringe Dispersion des gelösten Reagenzes sowie der aus dem Reagenz und dem Analyten gebildeten Verbindungen bzw. Komplexe aus dem Reaktionsvolumen heraus verhindert werden. Insbesondere wird so erreicht, daß das Reagenz bzw. das Reaktionsprodukt zu mindestens 90 %, vorzugsweise 95 % oder mehr zusammen mit dem Reaktionsvolumen in den Untersuchungsbereich 10 strömt bzw. gefördert wird.After the defined release or reaction, the device 12 releases the sample liquid 2, so that the sample liquid 2 - in particular the defined reaction volume - from the reaction region 9 in the study area 10 can continue to flow. Due to the measures mentioned, and in particular the means 16, a particularly small dispersion of the dissolved reagent as well as the compounds or complexes formed from the reagent and the analyte can in turn be prevented from the reaction volume. In particular, it is achieved that the reagent or the reaction product to at least 90%, preferably 95% or more flows together with the reaction volume in the examination area 10 or is promoted.

Im Untersuchungsbereich 10 wird dann die Probenflüssigkeit 2 bzw. das Reaktionsvolumen bedarfsweise nochmals temporär angehalten, um die jeweils gewünschte Untersuchung, insbesondere das beim Darstellungsbeispiel vorgesehene Binden von aus dem Reagenz und dem Analyten gebildeten Komplexen bzw. Verbindungen an einer im Untersuchungsbereich 10 vorzugsweise auf der Flachseite 4 immobilisierten Nachweischemikalie zu unterstützen. Jedoch ist ein solches temporäres Anhalten im Untersuchungsbereich 10 nicht unbedingt erforderlich, so daß die weitere, dem Untersuchungsbereich 10 zugeordnete Einrichtung 12 ggf. entfallen kann.In the examination area 10, the sample liquid 2 or the reaction volume is again temporarily stopped, if necessary, for the respectively desired examination, in particular the binding or compounds formed from the reagent and the analyte in the examination area 10, preferably on the flat side 4 immobilized detection chemical support. However, such a temporary stop in the examination area 10 is not absolutely necessary, so that the further, the examination area 10 associated device 12 may possibly be omitted.

Bei der vorgesehenen Nachweischemikalie handelt es sich insbesondere um eine immobilisierte Trockenchemikalie, beispielsweise einen Fängerantikörper, der die Verbindungen bzw. Komplexe aus Reagenz und Analyt fängt und dadurch bindet.The intended detection chemical is, in particular, an immobilized dry chemical, for example a capture antibody, which captures and binds the compounds or complexes of reagent and analyte.

Zusätzlich können weitere Untersuchungsschritte mittels anderer weiterer Chemikalien im Untersuchungsbereich 10 oder in mehreren nacheinander angeordneten Untersuchungsbereichen 10 durchgeführt werden. Beispielsweise kann auch eine Untersuchung dahingehend erfolgen, ob überhaupt der zu bestimmende Analyt in der Probenflüssigkeit 2 enthalten ist.In addition, further examination steps can be carried out by means of other further chemicals in the examination area 10 or in a plurality of examination areas 10 arranged one after the other. For example, an investigation can also be made as to whether the analyte to be determined is contained in the sample liquid 2 at all.

Der Untersuchungsbereich 10 schließt sich vorzugsweise unmittelbar an den Reaktionsbereich 9 an, so daß eine unerwünschte Dispersion des Reagenzes bzw, von dem Reagenz mit dem Analyten gebildeter Verbindungen oder Komplexe oder von sonstigen Reaktionsprodukten aus dem Reaktionsvolumen heraus in andere Bereiche der Probenflüssigkeit 2 zumindest im wesentlichen verhindert oder minimiert wird. Dadurch, daß sich der Untersuchungsbereich 10 vorzugsweise unmittelbar an den Reaktionsbereich 9 anschließt, werden nämlich das Totvolumen und damit die Dispersion minimiert.The examination region 10 preferably directly adjoins the reaction region 9, so that undesired dispersion of the reagent or of compounds or complexes formed by the reagent with the analyte or of other reaction products from the reaction volume into other regions of the sample liquid 2 is at least substantially prevented or minimized. Due to the fact that the examination area 10 preferably immediately adjacent to the reaction region 9, namely the dead volume and thus the dispersion are minimized.

Durch den flachen Querschnitt des Kanals 3 kann der Reaktionsbereich 9 und/oder der Untersuchungsbereich 10 verhältnismäßig kurz in Strömungsrichtung S ausgebildet werden, so daß insgesamt sehr kurze Strömungswege und dadurch eine geringe Dispersion erreichbar sind. Insbesondere ist der Reaktionsbereich 9 und/oder der Untersuchungsbereich 10 nur genauso lang wie oder kürzer als die Breite des Kanals 3 ausgebildet.Due to the flat cross section of the channel 3, the reaction area 9 and / or the examination area 10 can be formed relatively short in the flow direction S, so that a total of very short flow paths and thus a low dispersion can be achieved. In particular, the reaction area 9 and / or the examination area 10 is formed only as long as or shorter than the width of the channel 3.

Insbesondere ist die vorschlagsgemäß erreichbare Dispersion im Untersuchungsbereich 10 derart gering, daß die Konzentration des Reagenzes bzw. Reaktionsprodukts im Reaktionsvolumen im Untersuchungsbereich 10 um maximal 10 %, vorzugsweise weniger als 5 %, ganz bevorzugt höchstens 3 %, variiert.In particular, the dispersion which can be achieved according to the proposal in the examination zone 10 is so small that the concentration of the reagent or reaction product in the reaction volume in the examination zone 10 varies by a maximum of 10%, preferably less than 5%, very preferably at most 3%.

Die verhältnismäßig geringe Höhe H des Kanals 3 und die vorzugsweise vorgesehene Anordnung der Nachweischemikalie auf einer Flachseite, insbesondere der Flachseite 4, führen dazu, daß die im Reaktionsvolumen enthaltenen Komplexe bzw, Verbindungen aus Reagenz und Analyt oder sonstige Reaktionsprodukte sehr schnell bzw. mit einer sehr hohen Effizienz von der Nachweischemikalie o.dgl. gebunden werden können.The relatively low height H of the channel 3 and the preferably provided arrangement of Nachweischikalie on a flat side, in particular the flat side 4, cause the complexes contained in the reaction volume or, compounds of reagent and analyte or other reaction products very quickly or with a very high efficiency of the detection chemical or the like. can be tied.

In einem anschließenden Waschschritt strömt die Probenflüssigkeit 2 weiter in den Sammelbereich 11, der ggf. mit einem saugfähigen Material und/oder Leitelementen, wie den Erhebungen 19, versehen sein kann, um die Probenflüssigkeit 2 quasi aufzusaugen und einen Rückfluß zu verhindern, Das Volumen des Sammelbereichs 11 ist vorzugsweise mindestens um den Faktor 2 oder 5 größer als das Reaktionsvolumen, um ein effizientes Auswaschen von nicht gebundenem Reagenz, nicht gebundenen Reaktionsprodukten und/oder sonstigen eventuell störenden Partikeln oder Stoffen aus dem Untersuchungsbereich 10 zu erreichen.In a subsequent washing step, the sample liquid 2 flows further into the collecting area 11, which may optionally be provided with an absorbent material and / or guide elements, such as the elevations 19, to absorb the sample liquid 2 and to prevent a reflux, the volume of Collection area 11 is preferably greater than the reaction volume by at least a factor of 2 or 5 in order to achieve efficient washing out of unbound reagent, unbound reaction products and / or other possibly interfering particles or substances from examination area 10.

Anschließend erfolgt die Bestimmung des im Untersuchungsbereich 10 gebundenen Reagenzes oder Reaktionsprodukts vorzugsweise optisch, beispielsweise spektroskopisch. Dies ist insbesondere dadurch möglich, daß das Reagenz oder Reaktionsprodukt beispielsweise als Konjugat aus einem Antikörper und einem Farbkomplex, Farbpartikel o.dgl. aufgebaut ist. Aus der Anzahl oder Konzentration an gebundenen Komplexen bzw. Verbindungen kann dann insbesondere die Konzentration des Analyten in der Probenflüssigkeit 2 bestimmt werden. Folglich gestatten die vorschlagsgemäße Vorrichtung 1 und das voranstehend beschriebene Verfahren eine gegenüber herkömmlichen Testfilterstreifen o.dgl. wesentlich genauere Untersuchung der Probenflüssigkeit 2, insbesondere quantitative Bestimmung eines Analyten oder ggf. auch mehrere Analyte in der Probenflüssigkeit 2.Subsequently, the determination of the bound in the examination area 10 reagent or reaction product is preferably carried out optically, for example spectroscopically. This is particularly possible because the Reagent or reaction product, for example, as a conjugate of an antibody and a color complex, color particles or the like. is constructed. In particular, the concentration of the analyte in the sample liquid 2 can then be determined from the number or concentration of bound complexes or compounds. Consequently, the proposed device 1 and the method described above allow a comparison with conventional test filter strips or the like. much more accurate examination of the sample liquid 2, in particular quantitative determination of an analyte or optionally also several analytes in the sample liquid 2.

Claims (4)

  1. Use of a device (1)
    having a channel (3) which absorbs and conveys a sample liquid (2) by means of capillary forces and which has a flat side (4) over which the sample liquid (2) flows in a lateral and/or laminar manner,
    having a reaction area (9) having a dissolvable and/or reacting reagent on the flat side (4), wherein the reaction area (9) is completely fillable with sample liquid (2), thereby enabling reaction volume of sample liquid (2) to be defined,
    having a test area (10) which is downstream of the reaction area (9) and formed by the channel (3), wherein the reaction area (9) and/or the test area (10) are/is only exactly as long as or shorter than the width of the channel (3) and/or wherein the reaction area (9) and the test area (10) have at least substantially the same size, and
    having a means (16) for generating a slightly curved or straight-line flow front (F) of the sample liquid (2), wherein the means (16) is formed by the channel (3) being open at least longitudinally so that a lateral liquid stop for the sample liquid (2) is formed in the channel (3) and the sample liquid (2) is conducted in the channel (3) without any side walls,
    for testing a sample liquid (2),
    wherein, in order to dissolve and/or react the reagent, the reaction volume in the reaction area (9) and in the test area (10) is slowed down or temporarily stopped by an element (12, 15) and more than 90% of the reagent is dissolved in the defined reaction volume of sample liquid (2) and/or reacts therewith and, after the stoppage, more than 90% of the dissolved reagent and of a reagent product of the reagent flows together with the reaction volume into the test area (10).
  2. Use of a device (1) according to Claim 1,
    characterized in that the sample liquid (2) is conducted over guiding elements, preferably elevations (19), on the flat side (4) in order to achieve an at least substantially straight liquid front (F) and/or to prevent a lateral surge of the sample liquid (2).
  3. Use of a device (1) according to Claim 1 or 2,
    characterized in that the reaction volume is temporarily stopped in the test area (10), more particularly until at least 95% of complexes or compounds or reaction products formed from the reagent and an analyte in the sample liquid (2) are bound in the test area (10) by a detection chemical.
  4. Use of a device (1) according to any of the preceding claims,
    characterized in that the test area (10) is rinsed after the flow-through of the reaction volume with sample liquid (2) before a preferably optical test or determination is carried out, more particularly of reaction products, complexes, or compounds formed from the reagent and an analyte to be determined, which are bound to a detection chemical.
EP06001069A 2005-01-27 2006-01-19 Use of a device for analysing a liquid sample Active EP1685900B1 (en)

Applications Claiming Priority (1)

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DE200510003961 DE102005003961A1 (en) 2005-01-27 2005-01-27 Device for testing sample liquids, e.g. blood, plasma or urine, comprises a capillary channel, a reagent-containing zone with a device for holding liquid during reaction, and a test zone with a detection chemical

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EP1685900B1 true EP1685900B1 (en) 2011-03-30

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CN1811416A (en) 2006-08-02
DE502006009183D1 (en) 2011-05-12
ES2361169T3 (en) 2011-06-14
US20060216195A1 (en) 2006-09-28
ATE503578T1 (en) 2011-04-15
JP2006208388A (en) 2006-08-10

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