EP1931992A2 - Dispositif pour analyser des echantillons liquides - Google Patents

Dispositif pour analyser des echantillons liquides

Info

Publication number
EP1931992A2
EP1931992A2 EP06804339A EP06804339A EP1931992A2 EP 1931992 A2 EP1931992 A2 EP 1931992A2 EP 06804339 A EP06804339 A EP 06804339A EP 06804339 A EP06804339 A EP 06804339A EP 1931992 A2 EP1931992 A2 EP 1931992A2
Authority
EP
European Patent Office
Prior art keywords
rotor
sample container
lid
flow channel
recess
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP06804339A
Other languages
German (de)
English (en)
Inventor
Bernhard Ronacher
Christoph Reschreiter
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.)
CUBE DX GMBH
Original Assignee
ANAGNOSTICS BIOANALYSIS GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ANAGNOSTICS BIOANALYSIS GmbH filed Critical ANAGNOSTICS BIOANALYSIS GmbH
Publication of EP1931992A2 publication Critical patent/EP1931992A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
    • B01F27/27—Mixers with stator-rotor systems, e.g. with intermeshing teeth or cylinders or having orifices
    • B01F27/272—Mixers with stator-rotor systems, e.g. with intermeshing teeth or cylinders or having orifices with means for moving the materials to be mixed axially between the surfaces of the rotor and the stator, e.g. the stator rotor system formed by conical or cylindrical surfaces
    • B01F27/2722—Mixers with stator-rotor systems, e.g. with intermeshing teeth or cylinders or having orifices with means for moving the materials to be mixed axially between the surfaces of the rotor and the stator, e.g. the stator rotor system formed by conical or cylindrical surfaces provided with ribs, ridges or grooves on one surface
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5021—Test tubes specially adapted for centrifugation purposes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces

Definitions

  • the present invention relates to a device for the analysis of liquid samples.
  • high-through-put technologies Due to the enormous variety of different biomolecules and their different manifestations (chemical modifications) or variants (DNA sequences), high-through-put technologies (“high-through-put technologies”) are increasingly being used.
  • high-through-put technologies In addition to the classical methods of Northern and Southern blotting, in situ hybridization and any type of Western Blot techniques, multi-analyte detection systems have been increasingly developed in the last 15 years. The most important of these systems mostly use microtiter plates (eg as standard carriers for ELISA) with several hundred analyzes per run and on microarrays with more than 100 analysis points per cm 2 .
  • microtiter plates eg as standard carriers for ELISA
  • microarrays with more than 100 analysis points per cm 2 .
  • the binding partners are arranged, for example, in a specific pattern distributed over the carrier ("array"), wherein the carrier itself always has a planar geometry (eg US 5,800,992 and US 5,744,305) .
  • the respective analyte detection is based on a molecular binding of sample components Whether certain constituents are present in a sample can thus be recognized from the fact whether such components have entered into a bond with the immobilized binding partners.
  • the recognition of such a molecular bond takes place, for example, by means of an optical (eg fluorescence, luminescence, surface plasmon Resonance Spectrosco- py), an electrochemical or the measurement of a mass change of the molecules on the sensor.
  • incubation chambers are, inter alia, from the following suppliers: Corning, Genomic Solution, BioRad, Takara Bio, Advalytix, Genetix, Gesim, the incubation chambers are usually reusable and shell-shaped, with a planar support inside the chamber closed with sealing webs) can not be continued without serious consequences.
  • the result is an end point measurement, ie the measurement takes place after all incubation steps and provides one measured value per binding partner.
  • the situation is comparable to the application of the polymerase chain reaction and the subsequent analysis of the products and the corresponding further development of the real-time polymerase chain reaction.
  • This system is similar to the use of spatial supports that are used in a spherical shape as a stationary phase. Furthermore, it is known from WO 03/100401, WO 00/40334 and WO 02/08457 that a carrier in cylindrical form has immobilized binding partners in defined areas (spots) on its surface, and incubation of the sample liquid under immersion in a complementary sample container can perform tempered conditions.
  • the sample container is associated with a measuring device that can detect any interactions.
  • the devices described have, inter alia, the disadvantage that contaminations can be introduced into the sample container itself, or that it comes in the course of the investigation due to the usually small sample volumes that are used in an investigation, for the evaporation of the introduced into the device liquids. This results in changes in the concentration of the individual sample components, which can affect the measurements.
  • Another disadvantage of such devices in particular a device as described in WO 03/100401, is the unstable and not axially centered mounting of the rotor during its rotational movement.
  • a further object is to provide a device for analyzing samples which consist of a sample container and a rotor insertable therein, which enables improved storage of the rotor in the sample container.
  • This object is achieved by providing a device for analyzing liquid samples, with a rotationally symmetrical rotor insertable into a sample container, wherein an annular gap is provided between the sample container and the rotor, and the rotor has at least one flow channel for conveying liquids and / or gases into and / or from the interior of the sample container, wherein means for centering the rotor are provided on the rotor and optionally on the sample container.
  • the device disclosed herein comprises a rotor and a sample container. Between the rotor and the inner wall of the sample container, a radial annular gap is provided which defines the incubation space (the space in which binding partner and ligand can react or interact).
  • the rotor has a flow channel through which liquids can be conveyed into the sample container or removed from the sample container.
  • the means on the rotor and in the sample container for centered storage of the rotor in the sample container make it possible for the rotor to be stored in the sample container in a stable and centered manner. Thus, it is prevented that during the rotation of the rotor or the sample container, the rotor comes into contact with the sample container.
  • the centered bearing means on the rotor and sample container may be positively connected (e.g., bump-in, journal bearing sleeve, etc.).
  • At least one elevation is provided on the lateral surface of the rotor or on the inner wall of the sample container.
  • the arrangement of at least one elevation on the lateral surface of the rotor or on the inner wall of the sample container causes an improved mixing due to turbulence of the solutions present in the device according to the invention in the course of the rotation of the rotor or the sample container.
  • the survey may have various forms, but it is preferred to make the surveys so that a good mixing, without affecting the detection of binding between binding partner and ligand, is possible.
  • the rotor according to the invention has means for transmitting a torque to the rotor.
  • the sample container In order to set the rotor in rotation by a suitable device, means are provided on the rotor for enabling the transmission of torque to the rotor.
  • the sample container according to means which fix this radially. This prevents that during rotation of the rotor due to frictional effects and the sample container is set in rotation.
  • it is also to fix the rotor radially and to put the sample container itself in rotation.
  • the sample container has means for transmitting a torque to the sample container, and the means for transmitting a torque to the rotor serve as a fixing means.
  • radially fix means the provision of means used to prevent the rotor or sample container from rotating.
  • the means for transmitting a torque to the rotor is preferably an axially extending to the rotor longitudinal body.
  • the longitudinal body can have an arbitrary geometric plan, but an n-corner plan (n is selected, for example, selected from 3, 4, 5 and 6) is preferably used.
  • means may be provided on the longitudinal body, which facilitate the positive and / or non-positive connection with a torque-generating device (eg, the longitudinal body axial projections or projections or recesses or indentations have).
  • a cover for covering the interior of the sample container is provided on the rotor or on the sample container
  • a lid for closing or covering the sample container has many advantages. On the one hand, it is possible to keep the inside of the sample container contamination-free (for example, excluding dust and microorganisms), on the other hand, it is possible to minimize or completely eliminate evaporation processes within the sample container.
  • the lid may be provided such that it has at least or substantially a diameter as the opening of the sample container. If the diameter of the lid is greater than this opening, the lid protrudes beyond the edge of the sample container, the diameter is smaller or essentially the same as the sample container, the lid closes off the sample container within the container.
  • Another aspect of the present invention relates to a device for analyzing liquid samples, with a usable in a sample container rotationally symmetric rotor, wherein between the sample container and the rotor an annular gap is provided, the rotor at least one flow channel for the transport of liquids and / or gases in and / or from the interior of the sample container and at least one elevation is provided on the lateral surface of the rotor or on the inner wall of the sample container, wherein a lid for covering the interior of the sample container is provided on the rotor or on the sample container.
  • This device comprises a rotor, a sample container and a lid, which is the interior of the sample container in which the rotor in the inventive _ -j_
  • the upper portion of the rotor may either be itself a lid (e.g., by providing a protrusion on the rotor which can be inserted into the sample container or resting on the sample container) or the lid is attached to the rotor.
  • the rotor is provided with a flow channel through which the sample liquid and other liquids or gases required in the course of an analysis (for example incubation solutions, buffers, detection solutions, wash solutions, inert gas) are introduced into the interior of the sample container.
  • the flow channel thus provides a connection between the environment and the sample container interior.
  • the flow channel can be advantageously connected to a feed device (e.g., pipetting robot) for gases and liquids.
  • a feed device e.g., pipetting robot
  • the device according to the invention is also suitable for use in high-throughput screening.
  • the device can also be used as a disposable analysis device.
  • At least one elevation is provided on the rotor or on the inside of the sample container.
  • This survey serves to efficiently mix the solutions which are located in the radial annular gap between the rotor and sample container, since in the course of the radial movement of the rotor, the solution itself is set in radial motion and the proposed survey to Verwirbelun- gene in this solution leads, resulting in an efficient mixing of one or more solutions.
  • these surveys can be provided in a variety of arrangements.
  • the elevations may for example be arranged axially, spirally or in any other way along the rotor or the sample container.
  • Suitable analysis devices which can be provided with a lid according to the invention are disclosed, for example, in WO 03/100401.
  • a cover is applied to the device disclosed above according to the invention.
  • the "lid” (as a chanical barrier) the boundary of the sample container which closes off its opening on the side opposite the sample container bottom.
  • the lid comprises a material inert to the gases or liquids used in the apparatus and is substantially liquid and gas impermeable.
  • the lid has a diameter which is designed to introduce the lid into the sample container or to store it on the edge of the opening of the sample container.
  • the lid has a radial recess on the side of the lid facing the sample container, which has the same diameter as the sample container at the contact surface lid / sample container. This allows the lid to be stored on the sample container.
  • At least one opening may be provided on the lid. Without such an opening would be within the sample container during the introduction of liquids, if no suitable openings on the sample container or rotor are provided and the sample container interior air and liquid-tight configuration, form such a large pressure, thereby introducing more liquids is hardly possible , By an opening preferably provided on the lid, this can be prevented without significantly impairing the protective function of the lid.
  • the appropriate solutions e.g., sample solution, incubation solution, detection solution
  • means for centering the rotor in the sample container are provided on the sample container and on the rotor.
  • the sample container according to the invention and the rotor according to the invention have means for centered storage.
  • the invention relates to the means for centered mounting of the rotor in the sample container a directed into the interior of the sample container on the sample container bottom survey or indentation and the rotor a complementary recess.
  • This collection or indentation is used for radially movable bearings of the rotor in the sample container according to the invention, whereby a secure centering or positioning of the rotor in the sample container is made possible.
  • the protrusions may be of any shape (e.g., trunnion bushings). It is also quite possible to provide in the indentation or collection additional bearing elements, so that the rotor is mounted in the sample container by means of a ball bearing. Alternatively, it is also possible to introduce into the sample container bottom an insert which has a complementary indentation or elevation which is complementary to the elevation or indentation on the rotor. In such an embodiment, the sample container bottom could have no elevations or indentations.
  • a recess can be provided on the sample container bottom into which a complementary elevation located on the rotor or a complementary journal located on the rotor can be introduced for centered storage.
  • the elevation or indentation has a cylindrical shape, conical shape, truncated cone shape or a combined shape thereof.
  • the flow channel of the rotor is preferably connected to the means for centering the rotor connection, i. the storage means itself serves in this preferred embodiment itself as a flow channel.
  • Rotor On the inside of the means for centered storage of the Rotor is preferably provided at least one extending along the means and / or the flow channel recess.
  • the provision of such depressions along the means for centered storage and the flow channel is particularly advantageous when using the apparatus according to the invention for the analysis of small amounts of sample. Because of this depression, it is possible that the sample liquid can flow along this recess in the sample container. Furthermore, such a depression is advantageous if, for example, an elevation for supporting the rotor is provided within the sample device and on this elevation the complementary means on the rotor, rests in its function as a flow channel of the rotor. Without such a depression, this survey would make it difficult or prevent the sample liquid or other liquids from flowing into the sample container.
  • the receiving means or the flow channel can have at least one but also several such depressions (for example at least two, at least three, at least four, at least five, at least six, at least ten).
  • At least one recess is preferably provided, which permits a passage of liquid from the receiving means or flow channel of the rotor into the sample space (radial gap between the rotor and sample container) when the rotor is introduced entirely into the sample container.
  • This recess is vorzusgweise part of at least one recess in the flow channel of the rotor.
  • thermocouple can be used in the indentation provided in the sample container, the indentation must be designed accordingly to receive such elements
  • This thermocouple preferably consists of a cooling system (eg Peltier element) and a heating system (eg infrared radiator , Microwave radiator or Peltier element).
  • thermocouple can preferably der- be executed, that it allows the positioning of the device according to the invention in a measuring device or in a cartridge.
  • the indentation is thus preferably designed to accommodate a cooling and / or heating device.
  • the advantage of such an embodiment is further that in addition to the regulation of the temperature of the sample liquid and the device or the sample container according to the invention can be positioned.
  • the temperature regulation can also take place via the side wall or the bottom of the sample container (see for example WO 03/100401).
  • the indentation preferably has means for transmitting a torque to the sample container and / or means for fixing / positioning the sample container.
  • Such means for fixing the sample container or for transmitting a torque to the sample container is preferably a longitudinal body (e.g., pin or pin) provided axially in the recess, with a radial gap between the longitudinal body and the outside of the recess.
  • This longitudinal body can be inserted in a corresponding complementary part (e.g., recess, gripping device) in another device which can receive the device according to the invention.
  • a torque can not only be applied to the analysis device via the cover, but it is also possible to fix the cover together with the rotor and to set the sample container itself in rotation.
  • a detection system can be incorporated into the sample container or the receiving element (for example the means for centering the rotor in the sample container) of the container can include a detection system.
  • the receiving element for example the means for centering the rotor in the sample container
  • the means for centered storage is a magnetic bearing, wherein the magnetic bearing is preferably shown that magnets are provided on the rotor and optionally on the sample container.
  • Magnetic bearings allow the storage of the rotor in the sample container substantially without material contact by magnetic force.
  • a first magnet may be provided on the rotor, and the second magnet on the sample container or in another Vorrich- tion (eg, in which the device according to the invention is introduced for the measurement) or holder.
  • the magnets located on the rotor are preferably mounted annularly on the rotor bottom or its vicinity.
  • the magnets used in the rotor are preferably permanent magnets, and the magnets provided in the sample container or in the further apparatus may be not only permanent magnets but also electromagnets.
  • the sample container is preferably at least partially transparent.
  • the detection of the forming interactions between the binding partners and the ligands to be bound within the device according to the invention can take place in various ways.
  • the most frequently used principle for the determination of such interactions is the measurement of electromagnetic waves, in particular fluorescence, chemoluminescence, bioluminescence, fluorescence resonance energy transfer (FRET), which generates by binding corresponding marker molecules (eg labeled binding partners) to the ligands immobilized on the rotor by means of binding partners become.
  • FRET fluorescence resonance energy transfer
  • the molecules to be detected or with the molecules to be detected in competition with a corresponding substance are labeled (eg with a fluorophore or a quencher or groups of molecules that interfere with electromagnetic waves). If, for example, a DNA molecule is used, this can be done directly by incorporation of fluorophore-labeled nucleotides. On the other hand, if the detection is to take place indirectly via further secondary binding partners, it is also possible to label the molecules with markers such as, for example, biotin, digoxigenin (DIG).
  • DIG digoxigenin
  • the device according to the invention is preferably suitable for the measurement of chemiluminescence reactions in which the light emission of a substrate soluble in a corresponding enzyme (eg peroxidase) is measured.
  • a corresponding enzyme eg peroxidase
  • the detection of such a reaction can take place in different ways.
  • so-called CCD charge coupled device
  • CCD charge coupled device
  • the measurement can be carried out, for example, using the so-called TDI mode (Time Delayed Integration This mode requires a synchronization of the read-out speed of the CCD used and the movement of the object to be observed (eg microarray, biochip or rotor of the device according to the invention) (see WO 03/014400)
  • TDI mode Time Delayed Integration
  • the binding partners Preferably coupled to the rotor surface
  • these are synchronously rotated past the measuring system via the rotational speed as apparent lateral movement
  • photomultiplier arrays eg PMT ("photo multiplier tube"), APD ("avalanche photo diode"
  • PMT photo multiplier tube
  • APD avalanche photo diode
  • the measuring system eg photodiode array
  • the measuring system can be designed as a disposable product in combination with the sample container or can be a fixed component of the heating system located on the sample container. Since the rotor (binding partner carrier) and thus also the sample container preferably have a cylindrical shape, the requirement of a curved design of the photon sensor is of particular importance.
  • amorphous silicon diodes or photosensitive polymer layers as described for example in WO 03/015189 and WO 01/84644, are suitable.
  • silicon photodiodes in spatial proximity to the sample container surface is advantageous, above all, for the use of chemiluminescence detection methods, since these are used without an additional light source for exciting the light emission for the detection of the interactions between binding partner and ligands in the sample liquid can be.
  • Another possibility is, for example, the use of light sources, for example, in the interior of a recess which projects into the sample container.
  • the type of light source can Here, a direct, ie represent a unit with the carrier, or be an external light source, which is used for illumination or excitation of marker molecules in the sample liquid.
  • the detection systems described in US 2002/177144, WO 00/79326, WO 00/62549, WO 00/25113, WO 00/12759, WO 97/12030, US 5,585,639, US 2002/066865 and EP 0 947 824 are described, used for the device according to the invention.
  • the object of the continuous or stepwise (after changing time, liquid or temperature) optical representation of a mantle surface through a liquid film, which in turn is generated by the filling of the radial annular gap (see present invention and eg WO 03100401 A1), is solved according to the invention so that the observation (preferably the measurement of electromagnetic radiation emitted from the surface of said lateral surface) takes place through a transparent container, wherein the measuring system consists for example of a CCD camera or an APD (avalanche photo diode) or a comparable detection system , which is aligned substantially normal to the lateral surface and a strip surface of said lateral surface maps.
  • the measuring system is connected to the rotation unit in such a way that the rotation speed and the exposure time can be matched to one another.
  • the thus synchronized representation may optionally be incorporated into the use of the so-called TDI mode.
  • the stepwise rotation is synchronized in time with the integration time of the CCD chip.
  • the result is the line-dependent addition of the signal strength.
  • the measuring system may additionally include a lighting unit, which preferably consists of an opto-semiconductor (LED) and alternatively of at least one laser, a white light lamp, a gas discharge lamp, UV or IR lamp.
  • the measuring system can further be associated with a lens system consisting of at least one convergent lens, at least one filter set and / or an infrared filter.
  • the sample container can consist entirely of a material that is suitable and transparent for, for example, photometric measurement, or only partially (for example, providing viewing windows).
  • This viewing window (or the area in which the analyzer measures) may take the form of a lens (or lens structures) to optimize the optical properties. As a result, a focusing of the incoming and outgoing light and thus an increased yield of the measurement signal can be achieved.
  • the flow channel of the rotor is arranged axially.
  • the rotor In order to introduce sample or washing and incubation solutions into the sample container, the rotor has a flow channel through which liquids can be introduced into the device from outside.
  • the flow channel is preferably arranged axially on the rotor. It is of course also possible to provide the flow channel with an arbitrary course and any branches within the rotor and any exit point into the sample container.
  • an outlet opening In the lower region of the rotor, an outlet opening is preferably further provided.
  • the diameter of the flow channel of the rotor in the region of the sample container bottom is greater than in the region of the sample container opening.
  • the diameter of the flow channel of the rotor can increase in the direction of the sample container bottom (eg linear or stepped).
  • a linear or stepped enlargement can increase the storage area accordingly, which improves the concentric storage and consequently the measuring accuracy can.
  • increasing the diameter of the flow channel from initially, for example, 1-2 mm to 10-15 mm allows the introduction, such as described above, of a heating and / or cooling system in the storage means of the sample container.
  • the reduced wall thickness of the rotor reduces on the one hand the cost of materials and on the other hand can simplify and improve the manufacturing process (eg injection molding process: better injection and faster cooling).
  • the lid is detachably arranged.
  • the lid may be removably attached to the rotor, thereby making it possible, for example, to reuse the lid.
  • the cover fixed to the rotor or to provide the rotor and the cover in one piece.
  • the cover preferably has a flow channel of the rotor arranged in series and connected thereto flow channel.
  • the device After filling the device according to the invention with sample liquid or other liquids, the device can be closed by means of a lid which is applied to the rotor.
  • the lid of the rotor itself has a flow channel, which is in communication with the flow channel of the rotor. This makes it possible to prepare the device together with rotor and cover first and then introduce the introduced into the device liquids directly through the lid into the device.
  • the provision of a flow channel in the lid is particularly advantageous when using the device according to the invention as a flow cell.
  • the flow channel of the lid has a smaller diameter than the flow channel of the rotor.
  • the lid has a sealing lip at the edge.
  • the lid In order to efficiently seal the interior of the device, the lid has a sealing lip.
  • This sealing lip which extends over the entire outer edge of the lid, prevents the escape of liquid through the radial gap between the rotor and the sample container and additionally serves as a gas barrier which prevents the contact of sample liquid and e.g. Can prevent atmospheric oxygen.
  • This is particularly advantageous when examining samples which are sensitive to gases such as e.g. Oxygen, react and thus can falsify the measurement result.
  • the provision of a sealing lip on the cover allows a certain atmosphere to be provided within the device. For example, nitrogen may be introduced into the sample container, thereby making it possible to conduct analyzes under a protective atmosphere. Such studies are of particular interest when it comes to e.g. Analyze oxygen-sensitive samples.
  • the lid according to the invention which can be placed on a rotor according to the invention or is produced in one piece with it, has advantages in particular in contamination protection, in evaporation protection and in oxidation protection.
  • means are provided on the cover for transmitting a torque to the rotor connected to the cover.
  • a torque-generating device e.g., electric motor (stepping motor, DC motor, AC synchronous motor, AC induction motor), internal combustion engines, gas turbines, etc.
  • a torque-generating device e.g., electric motor (stepping motor, DC motor, AC synchronous motor, AC induction motor), internal combustion engines, gas turbines, etc.
  • the means for transmitting a torque to the rotor is preferably formed by an axially extending to the rotor longitudinal body.
  • the means which is responsible for the transmission of the torque to the rotor, may be designed differently, wherein preferably the means for transmitting a longitudinal body is.
  • a longitudinal body has the advantage that the device for generating a torque receives a point of attack on the lid, wherein the longitudinal body preferably has a polygonal shape (n is preferably selected from 3, 4, 5, 6 or 7).
  • n is preferably selected from 3, 4, 5, 6 or 7.
  • the rotor has fastening means for fastening the lid.
  • the rotor In order to fasten the cover both axially and radially on the rotor, the rotor has a fastening means on its upper area.
  • the fastening means can either be made in one piece with the rotor or the fastening means is applied to the rotor.
  • the fastening means are preferably formed by at least one radially arranged elevation.
  • the survey may have a tapering towards the free end diameter.
  • the elevation is provided with a helical groove (thread), with a radial recess or with a radial projection.
  • the cover preferably has a recess for receiving the fastening means, wherein according to a preferred embodiment, the recess itself has a helical groove (thread), a radial projection or a radial recess for fixing the fastening means of the rotor in the recess of the lid.
  • the recess itself has a helical groove (thread), a radial projection or a radial recess for fixing the fastening means of the rotor in the recess of the lid.
  • the elevation of the rotor which is used for fixing the lid, has a helical notch, a radial recess or a radial projection.
  • This makes it possible in a simple manner to provide a cover with a recess for receiving the fastening means of the rotor, wherein the indentation represents the counterpart of the survey and thus also has a helical notch, a radial recess or a radial projection.
  • the cover can preferably be fixed detachably on the rotor.
  • the lid is fixed to the rotor by a latching device.
  • the lid which has elevations or recesses on the contact surface with the rotor, engages on this, since corresponding complementary elevations or recesses are formed on the rotor.
  • At least one binding partner for binding at least one ligand is provided on the rotor and / or on the inside of the sample container, the at least one binding partner preferably having a biomolecule, in particular an antibody, antigen, hapten, peptide, polypeptides with various prosthetic groups, enzymes, hormones, a nucleic acid or derivatives thereof, such as peptide nucleic acids.
  • the device according to the invention is primarily suitable for the analysis of interactions between proteins, nucleic acids and other biological molecules, such as e.g. Membrane components, lipids, hydrocarbons and their derivatives, carbohydrates, haptens, hormones and synthetic agents (eg growth inhibitors such as antibiotics, pesticides, herbicides or fungicides, or inhibitors or activators of biochemical metabolic reactions), wherein at least one of the binding partners on a stationary phase in covalent or non-covalent form, or is bound as a sandwich via a general molecular adapter and brought into contact with the sample components to be analyzed via a liquid control system and their interactions are detected by means of a detection device.
  • proteins e.g. Membrane components, lipids, hydrocarbons and their derivatives, carbohydrates, haptens, hormones and synthetic agents (eg growth inhibitors such as antibiotics, pesticides, herbicides or fungicides, or inhibitors or activators of biochemical metabolic reactions)
  • growth inhibitors such as antibiotics, pest
  • biomolecules detected in a sample allow, for example, unambiguous conclusions as to the presence of viral, bacterial, eukaryotic, in particular animal and human DNA, or the occurrence of different forms of proteins (phosphorylations, glycosylations) or nucleic acids (methylation). or lipids with any glycosylations too.
  • binding partners are immobilized on the rotor or on the sample container.
  • the binding partners used in this case are selected according to the ligands to be bound.
  • biomolecules such as antibodies, nucleic acids, sugar chains, lectins or other proteins can be immobilized on the rotor in order to bind ligands (which likewise constitute binding partners) to the immobilized binding partners.
  • the ligand can also be bound to the rotor or sample container, so that in this case the ligand is by definition (in the context of the present invention) referred to as a binding partner.
  • the device according to the invention is suitable not only for the detection of certain substances in a sample, but also for the conversion of substances, which can record, for example, enzyme kinetics. Furthermore, the device of the invention can be used to carry out enzyme reactions, e.g. DNA polymerase reactions ("solid phase amplification", "APEX"), DNA ligase reactions, DNA methyltransferase reactions, restriction endo and exonuclease reactions, oxidoreductases, hydrolases, Ligase, lyase, isomerase, phosphatase, kinase, methylase and transferase reactions.
  • enzyme reactions e.g. DNA polymerase reactions ("solid phase amplification", "APEX"), DNA ligase reactions, DNA methyltransferase reactions, restriction endo and exonuclease reactions, oxidoreductases, hydrolases, Ligase, lyase, isomerase, phosphatase, kinase
  • the rotor preferably has at least one outwardly directed and / or the inner jacket of the sample container at least one inwardly directed radial projection as a spacer element.
  • the rotor may have at least one, preferably at least two, more preferably at least three, most preferably at least four, in particular at least five, radially outward. have directional spacers. Preferably, at least one radially outwardly directed spacer element at the bottom and / or top of the rotor.
  • a radial projection which may also comprise continuously the lateral surface of the rotor, is particularly well suited to be used as a spacer element.
  • the spacer also serves to limit the sample or incubation space.
  • the radial projection preferably has, in particular along the recess in the interior of the flow channel, at least one recess.
  • the recess on the radial projection makes it possible for the solutions introduced through the flow channel to flow outward from the interior of the rotor into the reaction space between the rotor and the sample container wall.
  • the recess may also be such that the diameter of the radial projection is reduced.
  • radial projections can also be provided on the inside of the sample container.
  • the protrusions on the sample container perform the same job as the radial protrusions on the rotor.
  • the radial projections are preferably provided on the sample container.
  • the projections can also serve as a "bubble slide", which can take any resulting or introduced air bubbles or dirt particles from the projections. The result is a distance up or down, or a uniform load on the entire surface and not just an area, as is the case with planar carriers.
  • the radial projection of the sample container has at least one recess, in particular an opening.
  • the sample container has means for its radial fixation in a cartridge.
  • lent rotor or in a cartridge (eg for transport) to fix can be provided on the sample container, for the radial fixation, a means.
  • the means for radial fixation of the sample container in the holder or cartridge is preferably at least one projection provided on the sample container bottom, in particular a knob.
  • the provision of a projection on the sample container bottom makes it possible in a simple manner to fix the sample container in a cartridge or device holder if these have corresponding means which can receive this projection.
  • the cartridge according to the invention which may or may not be part of a measuring device, thus serves to receive the analytical device according to the invention (sample container, rotor and lid).
  • the cartridge is thus a transport and / or storage unit for the analysis device.
  • the rotor, the sample container and / or lid is formed from a plastic, this preferably a cyclo-olefin copolymer, polystyrene, polypropylene, polyethylene, Acetatpo- lymer, acrylonitrile butadiene styrene, polymethyl methacrylate, PVC , Polyethylene terephthalate, polytetrafluoroethylene or a combination thereof.
  • plastic preferably a cyclo-olefin copolymer, polystyrene, polypropylene, polyethylene, Acetatpo- lymer, acrylonitrile butadiene styrene, polymethyl methacrylate, PVC , Polyethylene terephthalate, polytetrafluoroethylene or a combination thereof.
  • plastic this preferably a cyclo-olefin copolymer, polystyrene, polypropylene, polyethylene, Acetatpo- lymer, acryl
  • the surface of the rotor is preferably coated with a metal, preferably a semiconducting metal, a polymer, silicon or a silicon compound with carbon, preferably with graphite, DLC (diamond-like-carbon) or diamond, or a combination thereof, the metal preferably being gold , Palladium, silver or a combination thereof and the silicon compound is silica.
  • a metal preferably a semiconducting metal, a polymer, silicon or a silicon compound with carbon, preferably with graphite, DLC (diamond-like-carbon) or diamond, or a combination thereof, the metal preferably being gold , Palladium, silver or a combination thereof and the silicon compound is silica.
  • a metal preferably a semiconducting metal, a polymer, silicon or a silicon compound with carbon, preferably with graphite, DLC (diamond-like-carbon) or diamond, or a combination thereof, the metal preferably being gold , Palladium, silver or a combination thereof
  • SoIGeI Dissolved cathalytic Si-oxide particles harden to a glass layer over the plastic surface
  • Chemical modification by introduction of chemically reactive groups, epoxy, aldehydes or plasma treatment to activate the surface layer of the plastic and introduction of reactive chemical groups (amino, hydroxyl, epoxy, aldehyde, carboxy groups) are preferred surface coating methods or modification.
  • molecules are bound to the surface via sulfur compounds (Au-S-R) in combination with a second reactive group as the residue, preferably an NHS ester or maleimide or similar reactive groups.
  • a second reactive group as the residue, preferably an NHS ester or maleimide or similar reactive groups.
  • the corresponding modification of the surface occurs via a chemical immersion process in a solution of dissolved DMSO solution. It is then washed with water or apolar solvent, MeOH or acetone or preferably with isopropanol. After drying, the biological binding partners (e.g., antibodies, proteins, peptides, modified DNA, or generally organic molecules which themselves carry reactive groups for covalent immobilization) are bound to the surface.
  • Another aspect of the present invention relates to a cartridge for receiving an analysis device according to the invention, wherein the cartridge has an opening for introducing the sample container and a side boundary provided with a recess, wherein the recess for axial fixing of the rotor is formed.
  • a cartridge can serve to protect the sample container including the rotor and lid from mechanical stress and at the same time serve as packaging.
  • the cartridge can be designed such that it can be used for automatic filling, for example via a magazine with a plurality of cartridges, and for unambiguous positioning in a device in which the device according to the invention can be used. position (eg by means of a gripper arm).
  • the cartridge also preferably has a side boundary provided with a recess, wherein the recess is designed to receive the upper region of the rotor or the means provided on the cover for transmitting a torque to the rotor and to support it in a radially immovable manner.
  • the side boundary provided in the cartridge with a recess has the advantage that this recess can either receive the upper region of the rotor and the cover is thus located above the lateral boundary, or that this recess can receive the means for transmitting the torque provided on the cover but without disturbing the radially movable bearing.
  • This makes it possible to provide the rotor in different positions within the sample container. Such positioning is advantageous, for example, in an ideal incubation.
  • the sample liquid may be brought to a certain temperature (e.g., for hybridization) prior to use in the apparatus of the present invention, or a sample conditioning step is accomplished, which is accomplished by temperature differences (e.g., PCR). At this time, no contact with the binding partners on the rotor surface may take place.
  • the side boundary of the cartridge (as a horizontal plate) is preferably provided over the entire depth with a recess which tapers at the edges such that the radial groove of the rotor can be inserted with some pressure and a proposed constriction.
  • the boundary (bottom of the cartridge) opposite the side boundary provided with the recess preferably has a recess for receiving the means for radial fixation of the sample container in the cartridge.
  • this recess / depression which is substantially complementary to the means provided on the sample container bottom means for radial fixation, the radial movement of the sample container is prevented.
  • the recess may be formed as a guide rail, which allows the recording of the means for radial fixation of the sample container.
  • Another aspect of the present invention relates to a rotor for a device according to the invention.
  • Another aspect of the present invention relates to a sample container for a device according to the invention.
  • kits for analyzing samples comprising:
  • the kit according to the invention is particularly well suited for the analysis of samples, i. for the determination of ligands in samples, or for carrying out chemical and biochemical reactions in the interior of the devices according to the invention.
  • Another aspect of the present invention relates to a flow cell for analyzing samples comprising:
  • the devices of the invention are also particularly well suited for use as flow cells.
  • Flow cells are used in many areas of analysis.
  • flow cells are suitable when it comes in the course of an analysis to a repeated change of sample liquids, incubation liquids, washing liquids and detection fluids.
  • the flow channel of the rotor serves as Einströmungsort, which is located at the upper end of the rotor according to the invention.
  • the outflow location at which the liquids located in the sample container can be removed therefrom can be provided on the sample container or on the lid.
  • the flow cell itself, as well as the measuring cell of a device according to the invention, not as Flow cell is adapted, defined by the annular gap between the rotor and the sample container. In such an embodiment, it is quite possible to dispense with the lid according to the invention.
  • a flow cell according to the invention can be used for example for the investigation of larger amounts of sample liquid such as water or other samples (eg waste water, cell culture supernatants), which is moved through the flow cell and thus brought into contact with the preferably attached to the rotor binding partners.
  • sample liquid such as water or other samples (eg waste water, cell culture supernatants)
  • the measurement can take place in real time, ie, during the flow through the sample liquid through the flow cell, the sample is examined by detecting the binding between the substance to be detected and its binding partner, which is immobilized in the device.
  • Another field of application of the flow cell according to the invention is the flow of culture medium through this cell to provide nutrients to growing cells on the rotor and to monitor their response to changes in the culture medium and optionally to analyze, fix and stain the cells.
  • At least one opening for the discharge of liquid is provided on the rotor, preferably in a spacer element or in the lid, preferably adjacent to the sealing lip, or on the sample container.
  • the liquid introduced through the flow channel can be efficiently removed from the device. It is thus not necessary to use the flow channel both for the introduction and for the removal of liquids into or out of the sample container.
  • the device according to the invention can be used for numerous uses, methods and methods. These uses include the detection of substances in a sample, the performance of enzymatic reactions in the device (e.g., amplifications of nucleic acids), etc.
  • One particular method for which the device of the present invention is particularly useful is the detection of substances in saliva.
  • the aim of such a method is the detection of eg illicit substances (drugs) in saliva. In doing so, additional next a saliva decrease in a withdrawal liquid. The removed saliva undergoes no purification (filtration, centrifugation or similar methods). The saliva-containing fluid thus obtained is used directly in the device according to the invention.
  • the following steps are preferably carried out, it being understood that the number can be varied according to the type of steps.
  • the antibody to the desired substance e.g., antigen
  • a labeled antigen o. in dried form after spraying onto the inner surface of the sample container
  • the bound antibodies can be detected by their fluorescence labeling.
  • the collected light intensity is linearly proportional to the amount of fluorophores and thus proportional to the amount of bound antibodies.
  • a quantitative statement about the amount of competitors (drugs) is thus possible. It has surprisingly been found that non-purified (ie not centrifuged and filtered) saliva in combination with the microarrays located on the rotor can be used directly. It is advantageous that thus eliminating purification steps and thus a mobile use (eg in traffic control, directly in the infirmary) is possible because mobile centrifuges with the required G-number (of more than 2000g) are hardly feasible.
  • the variation of the measurement results can be minimized within the device (about 2-7%).
  • the detection of nucleic acids can be carried out in a sample.
  • the sample is pretreated after collection (e.g., blood) and the nucleic acid pre-purified.
  • the detection takes place via the fluorescence intensity measurement of the array on the carrier (rotor).
  • the signal comes from fluorophores that are in adsorbent or covalent contact with the immobilized molecules of the rotor array. Two signal generation methods are used:
  • Fluorescence Enegrie Transfer to quenching dyes, or other fluorophores
  • Both methods have in common that they require an enzymatic reaction to reach a detectable measurand.
  • this will be the DNA dependent DNA polymerase (preferably Taq polymerase).
  • the process is commonly referred to as Solid Phase PCR (Polymerase Chain Reaction) or Solid Phase Amplification.
  • Solid Phase PCR Polymerase Chain Reaction
  • Solid Phase Amplification The whole process of a Analysis is planned as follows (example infection diagnostics, pathogen-specific DNA primers are spotted on the rotor surface as an array)):
  • the sample e.g., blood
  • the sample is taken from the patient.
  • the DNA of the sample (s) is prepared accordingly.
  • the sample is denatured (94 0 C), while the double strands separate into single DNA strands.
  • the device is rotated at 72 0 C and incubated: this involves the polymerization (chain extension) of the primer due to the hybridized DNA sample molecules.
  • the sample is again denatured (94 0 C), the double strands separate again.
  • This duplex formation is measured by fluorescence measurement (e.g., FRET or CYBRgreen).
  • Steps 4-9 are repeated cyclically until a meaningful signal can be expected.
  • the above illustration of the analysis process makes it clear that on the one hand a spatially resolved measurement is required to implement a CIS-PCR array and on the other hand it must be able to repeat this measurement any number of times without changing the analytical conditions (eg by drying the solid phase). Moreover, the measuring system must be able to change the temperature of the sample according to an adjustable profile at any time.
  • the device according to the invention is the only technology known to us which meets all these requirements. corresponds gen and thus makes the ansich known method the first time safely applicable.
  • the device according to the invention is suitable for the direct reverse transcriptase PCR on a solid support (eg US Pat. No. 6,844,158), determination of DNA polymorphisms (eg DE 10 160 983), determination of DNA sequences by means of parallel amplification (eg EP 1 186 669), solid-phase PCR (eg JP 2001 299 346, US Pat. No.
  • the analysis device in which the device according to the invention can be used to analyze samples may comprise a plurality of components, wherein in particular at least one rotation and detection device are required.
  • the analyzer may include a light source (e.g., laser box), an incubation chamber, a fluid change device (optionally coupled to a rotating device), a plurality of fluid reservoirs, and a cartridge.
  • a typical analyzer may have the following components:
  • a laser box Place the laser (up to 3 different laser types in parallel) or light sources that come out of the box bundled and centered
  • An incubation chamber location of temperature control, rotation, fluid change and measurement
  • a rotation and fluid exchange device serves to accommodate the device according to the invention on the lid, ensures the rotation, the transport in the device thus also the positioning, the liquid exchange (supply and discharge)
  • a detector e.g. a CCD camera, alternatively also avalanche photodiodes (APD)
  • Liquid reservoirs they are under controlled positive and negative pressure, which, in combination with point 3. and valves placed there, enables the supply and discharge of liquids.
  • a magazine location of the device according to the invention, before and after the measurement to eg 8 pieces per magazine from the device can be managed.
  • FIGS. 1A and 1B show two spatial views of a rotor according to the invention.
  • Figs. 2A and 2B show a side view of the rotor.
  • Fig. 3 shows a plan view of the rotor.
  • Fig. 4 shows a cross section of the rotor (section A-A of Fig. 3).
  • Fig. 5 shows a perspective view of a sample container according to the present invention.
  • Fig. 6 shows a side view of the sample container.
  • Fig. 7 shows a cross section of the sample container (section A-A of Fig. 6).
  • Fig. 8 shows a bottom view of the sample container.
  • Fig. 9 shows a three-dimensional view of a lid according to the invention.
  • Fig. 10 shows a side view of the lid.
  • Fig. 11 shows a cross section of the lid (section A-A of Fig. 10).
  • FIG. 12 shows the enlargement of detail B from FIG. 10.
  • Fig. 13 shows a top view of the lid.
  • Fig. 14 shows a perspective view of a cartridge according to the invention.
  • Fig. 15 shows a side view of the cartridge.
  • Fig. 16 shows a cross section of the cartridge (section A-A of Fig. 15).
  • Fig. 17 shows a cross section of the cartridge (section B-B of Fig. 15).
  • Fig. 18 shows a cross section of the cartridge (section C-C of Fig. 15).
  • Fig. 19 shows a plan view of the kart search.
  • Fig. 20 shows a perspective view of a device according to the invention with sample container, rotor and lid.
  • Fig. 21 shows a side view of a device according to the invention with sample container, rotor and lid, wherein the broken lines represent edges not visible from the outside.
  • Fig. 22 shows a cross section of a device according to the invention with sample container, rotor and cover (section AA of Fig. 21).
  • Fig. 23 shows a side view of a device according to the invention with sample container, rotor and lid.
  • Fig. 24 shows a further side view of a device according to the invention with sample container, rotor and lid.
  • Fig. 25 shows a perspective view of a device according to the invention with sample container, rotor and lid.
  • FIG. 26 shows a three-dimensional view of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention.
  • FIG. 27 shows a further spatial view of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention.
  • Fig. 28 shows a side view of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention, wherein the broken lines represent edges not visible from the outside.
  • Fig. 29 shows a cross section of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention (section A-A of Fig. 28).
  • FIG. 30 shows a further side view of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention.
  • Fig. 31 shows a cross section of a device according to the invention with sample container, rotor and lid in a cartridge according to the invention (section A-A of Fig. 30).
  • FIG. 32 shows the coefficients of variation of measurements with the device according to the invention, in which the rotor surface was spotted with antibody (specifically against mouse antibodies) and incubated with mouse monoclonal antibodies (fluorescently labeled with Dylight 547). After incubation, the rotors were washed with PBS 0.5% Tween 20 and scanned.
  • FIGS. 33 and 34 show the maximum signal level of measurements with the device according to the invention, in which the rotor surface was spotted with antibody (specifically against mouse antibodies) and incubated with mouse monoclonal antibodies (fluorescently labeled with Dylight 547). After incubation, the rotors were washed with PBS 0.5% Tween 20 and scanned. The measurement series corresponds to individual measurements with different incubation times. The maximum reached after about 30 minutes becomes clear in the measurement series with rotating incubation. The static incubation does not reach the signal level or a stagnation of the signal increase even after 90 minutes, ie 3 times the incubation time. Reproducible measurements, especially for single measurements, which are currently common, depend on the achievement of an equilibrium (stagnating, flattening signal height).
  • FIG. 35 shows the dependence of the signal height on the incubation period during a rotating incubation.
  • FIG. 36 shows the recording of a marked spot located on the rotor with rotating centered (A) and uncentered (B) mounting of the rotor.
  • Fig. 37 shows the relative focusing errors in a rotary incubation with the device according to the invention in comparison with a device which has no means for centric storage.
  • Fig. 38 shows the recording of the signals as a function of time (binding kinetics).
  • a cylindrical rotor 1 according to the invention with a fastening means 2 comprising a radial recess 3 for fixing a lid is shown.
  • the rotor 1 has an outwardly directed radial projection 4 at the upper 3a and 3b lower region.
  • elevations 6 are provided, which serve for better mixing of the sample container located between the rotor 1 and the sample liquid.
  • the rotor 1 also has a flow channel 7, which has a smaller diameter in the upper region 3a of the rotor 1 in the attachment means 2 than in the lower region 3b of the rotor 1.
  • two recesses 8 extending along the flow channel 7 are provided Transport of a liquid along the flow channel 7 serve.
  • Figs. 2A and 2B show another side view of the rotor
  • a recess 10 is provided, which is a liquid passage from the flow channel 7 in the sample space (radial gap between the rotor 1 and sample container 11) at wholly in the sample container
  • 3 is a plan view of the rotor 1, in which the inner boundary 9 of the flow channel 7 in the rotor 1 is shown by broken lines.
  • Fig. 4 is a cross section (section A-A, Fig. 3) of the rotor 1 is shown.
  • the flow channel 7 of the rotor 1 has a wider diameter in the lower region 3b than in the upper region 3a. Furthermore, the flow channel 7 in the lower region 3b is cylindrical and in the region of the diameter taper truncated cone-shaped. In the fastening means, the flow channel is again cylindrical.
  • FIG. 5 shows the spatial representation of a sample container 11 which is designed to receive the rotor 1.
  • the sample container 11 has on the side boundary 12 a transparent viewing window 13, through which the signals generated in the device according to the invention can pass substantially unhindered.
  • the transparent viewing window 13 can be adapted according to the measuring system used and, for example, have a curvature.
  • two projections 15 for example in the form of buttons
  • the sample container 11 has a radial projection 16 at its opening for receiving the rotor 1.
  • Fig. 6 shows a side view of the sample container 11 (see Fig. 5). The broken lines indicate elements in the interior of the sample container, which are shown in more detail in Fig. 7.
  • Fig. 7 the cross section (section A-A) of the sample container 11 is shown.
  • a cylindrical recess 17 is provided with a conical end portion 18, the shape of which is formed so as to support the rotor 1 according to the invention radially movable.
  • a cylindrical longitudinal body 19 is provided in the indentation 17, which can serve to position the sample container 11.
  • a cooling and / or heating device can also be introduced.
  • FIG. 9 shows a cover 20 according to the invention, which can be fixed to the rotor 1, in particular to its attachment means 2 (see FIGS. 2A and 2B).
  • the lid 20 has a sealing lip 21, which serves to seal the interior of the sample container liquid and gas-tight. Further, an opening provided with an opening indentation 22 for receiving the fastening means 2 of the rotor 1 is applied to the lid.
  • a cylindrical hollow body 23 is provided, which can be used both as a flow channel for introducing liquids into the sample container 11 and for transmitting a torque from a torque-generating device to the rotor 1.
  • axially extending elevations 24 may be provided which improve the grip of the hollow body 23.
  • Fig. 10 shows a side view of the lid 20, whose cross-section is shown in Fig. 11.
  • a radial projection 25 is provided, which can engage in the recess 3 of the fastening means 2 of the rotor 1.
  • a thread is also provided in the indentation 22 in order to enable the fixing of the cover 20 to the rotor 1.
  • the detail B from FIG. 11, which comprises the sealing lip 21, is shown enlarged in FIG. 12.
  • the lid 20 has immediately adjacent to the sealing lip 21 at least one opening 26 through which either escaping gas or, in the case of a flow cell, leaking liquid can be removed from the sample container.
  • Fig. 13 illustrates a plan view of the lid 20.
  • Fig. 14 is a perspective view of the cartridge 27 according to the invention, which is designed to receive the device according to the invention comprising rotor 1, sample container 11 and lid 20 is shown.
  • the cartridge has a horizontal plate 28 (side boundary), which is provided over the entire depth with a slot which is designed around the upper portion 3a of the rotor 1 or provided on the cover 20 means 23 for transmitting a torque to the rotor take up and store radially movable.
  • a guide groove (recess) 30 is inserted, which is substantially complementary to the elevations 15 located on the sample container 11 and prevents the radial rotation of the sample container 11 when the device is inserted.
  • the horizontal plate 28 is designed so that the pressure, which must be automatically expended by the placement of the lid 20 by hand or later, is recorded. Also provided on the bottom 29 of the cartridge is an outwardly directed projection 31 which serves to stabilize the cartridge 27 on a support surface.
  • FIGS. 15 to 19 show various sectional views of the cartridge 27.
  • FIG. 20 shows the spatial view of a device according to the invention comprising a rotor 1, a cover 20 and a sample container 11.
  • the rotor 1 is in this figure entirely in the sample container 11 (so-called "measuring position") - To a measurement using To perform the device according to the invention, the rotor 1 must be in this position during the measurement.
  • Fig. 21 shows a side view of the device according to the invention, in which the edges not visible from the outside are shown with broken lines.
  • Fig. 22 shows a cross section of the device according to the invention.
  • the sealing lip 21 of the lid 20 presses in the "measuring position" against the inner wall of the sample container 11 and thereby forms a closed measuring space, which may be separated by one or more openings, e.g.
  • the supply and the discharge of liquids or gases for example, sample liquid, buffer, detection liquid, washing liquid
  • a radial annular gap 32 is provided between the inner wall of the sample container 11 and the outer wall of the rotor 1.
  • FIGS. 23 and 24 show further side views of the device according to the invention.
  • Fig. 25 is another spatial view of the device according to the invention.
  • the indentation 17 of the sample container 11 is visible, which may optionally at least partially serve as a bearing surface for the rotor 1.
  • the elevations 15 on the sample container bottom serve for radially fixing the sample container 11 in a cartridge 27.
  • FIGS. 26 and 27 spatially illustrate the device according to the invention in a cartridge 27.
  • the elevations 15 on the sample container bottom are located in the guide groove 30th in the bottom 29 of the cartridge.
  • the position in which the device according to the invention is located in the cartridge 27, can be referred to as "storage position", since the rotor 1 is not completely in this position in the sample container 11 and thus no annular gap 32 can be formed for a measurement would be needed.
  • the rotor 1 due to the friction of the upper portion 3a of the rotor 1, the Befest Trentsgffens 2 and / or the cover 29 with the slot of the horizontal plate 28 and with the horizontal plate 28 itself, for fixing the device according to the invention is used in the cartridge 27, the rotor 1 is not offset with a sufficient torque.
  • the slot of the horizontal plate 28 serves to receive the fastening means 2 of the rotor 1.
  • the horizontal plate 28 is thus surrounded by the cover 20 on one side and the rotor 1 on the other side.
  • Example 1 Variation coefficient of measurements with: the device according to the invention with static and rotating incubation
  • Table 1 Scanning settings were 0.9 7 300. The devices were incubated for 5 min with 1 ⁇ l of antibody (1 mg / ml) labeled with Dylight 547 nm rotatingly or statically incubated, evaluation took place with a diameter of 200 ⁇ m.
  • test protocol which serves for the detection and quantification of morphine in saliva with the device according to the invention:
  • Example 2 Comparison of the signal height of measurements with the device according to the invention in static and rotating incubation
  • Table 2 Scanning settings were 0.9,7,300. The devices were rotary or static incubated with 1 ⁇ l of antibody ( ⁇ g / ml) labeled with Dylight 547nm. Evaluation was done with 200 ⁇ m diameter. The morphine BSA signal was evaluated.
  • FIGS. 36A and B it can clearly be seen that problems arise with the recording of the measurements in the case of a noncentric bearing of the rotor in the sample container.
  • the course of the focusing error over the cylinder jacket is shown in FIG. It can be seen that the image of the markings change depending on the pendulum movement over the circumference from sharp to blurred.
  • the centric storage of the device according to the invention shows a regular sharpness with a small fluctuation range (calculation tolerance).
  • FIG. 38 clearly shows an increase of the signal (average of 9 identical spots on the rotor surface of the device according to the invention) as a function of the incubation time.
  • the experiment was carried out according to the protocol of Example 1, wherein measurements were taken during incubation with the labeled antibody without removal of the unbound antibody by washing were carried out. The uniformly elevated background (labeled antibody in the solution) was subtracted from the signal level. The remaining signal was included as a value in the table. The signal height reaches the maximum after 13 minutes. The experiment was ended.
  • binding kinetics or dissociation events can be created but also competitions

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  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne des dispositifs pour analyser des échantillons liquides, comprenant un rotor (1) à rotation symétrique pouvant être inséré dans un récipient pour échantillon (11), un espace annulaire (32) étant disposé entre le récipient d'échantillons (11) et le rotor (1) et le rotor présentant au moins un canal d'écoulement (7) destiné à acheminer des liquides et/ou des gaz à l'intérieur et/ou à l'extérieur de la chambre interne du récipient pour échantillon (11). Des organes destinés à centrer le rotor(1) sont placés sur le rotor et éventuellement sur le récipient pour échantillons (11) et/ou un couvercle est placé sur le rotor.
EP06804339A 2005-10-07 2006-10-09 Dispositif pour analyser des echantillons liquides Withdrawn EP1931992A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AT0164105A AT502549B1 (de) 2005-10-07 2005-10-07 Vorrichtung zur analyse von flüssigen proben
PCT/AT2006/000411 WO2007041734A2 (fr) 2005-10-07 2006-10-09 Dispositif pour analyser des echantillons liquides

Publications (1)

Publication Number Publication Date
EP1931992A2 true EP1931992A2 (fr) 2008-06-18

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EP06804339A Withdrawn EP1931992A2 (fr) 2005-10-07 2006-10-09 Dispositif pour analyser des echantillons liquides

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Country Link
US (1) US7927546B2 (fr)
EP (1) EP1931992A2 (fr)
AT (1) AT502549B1 (fr)
AU (1) AU2006301950B2 (fr)
CA (1) CA2624280C (fr)
WO (1) WO2007041734A2 (fr)

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Also Published As

Publication number Publication date
CA2624280C (fr) 2015-12-08
AT502549B1 (de) 2007-06-15
CA2624280A1 (fr) 2007-04-19
AU2006301950B2 (en) 2011-09-08
US20080233590A1 (en) 2008-09-25
US7927546B2 (en) 2011-04-19
AT502549A1 (de) 2007-04-15
WO2007041734A3 (fr) 2007-06-21
AU2006301950A1 (en) 2007-04-19
WO2007041734A2 (fr) 2007-04-19

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