WO2002039096A1 - Analyse de dosages biologiques et biochimiques - Google Patents

Analyse de dosages biologiques et biochimiques Download PDF

Info

Publication number
WO2002039096A1
WO2002039096A1 PCT/GB2001/004933 GB0104933W WO0239096A1 WO 2002039096 A1 WO2002039096 A1 WO 2002039096A1 GB 0104933 W GB0104933 W GB 0104933W WO 0239096 A1 WO0239096 A1 WO 0239096A1
Authority
WO
WIPO (PCT)
Prior art keywords
array
flow cell
sample
light
substrate
Prior art date
Application number
PCT/GB2001/004933
Other languages
English (en)
Inventor
Nikolai F. Schwabe
Linda C. Tan
Original Assignee
Proimmune Limited
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 Proimmune Limited filed Critical Proimmune Limited
Priority to AU2002212520A priority Critical patent/AU2002212520A1/en
Publication of WO2002039096A1 publication Critical patent/WO2002039096A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • G01N21/6454—Individual samples arranged in a regular 2D-array, e.g. multiwell plates using an integrated detector array
    • 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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
    • B01L9/523—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for multisample carriers, e.g. used for microtitration plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/251—Colorimeters; Construction thereof
    • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • 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
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0819—Microarrays; Biochips
    • 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
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0877—Flow chambers

Definitions

  • the present invention describes an apparatus and method for analysing a biological or biochemical assay.
  • Samples to be analysed may form an array on a substrate or be located within a flow cell.
  • the advantage of the array-based technologies is their potential scalability for analyzing a high number of probes within, a short amount of time. As such they represent an extension of multi-well-plate based assays performed by scientists largely manually in the past.
  • the interest in large-scale substrate of chip based array technology has been fueled by the development of techniques of producing such arrays efficiently using lithographic or other techniques.
  • a method for making very large arrays of polymer sequences, including polypeptides and polynucleotides on very small substrates has been disclosed, for example, in US 5,143,854, the disclosure of which is incorporated herein by reference.
  • Analysing the arrays thus produced normally requires the performance of one or several biochemical reactions followed by optical imaging performed on the array.
  • the imaging has typically been achieved either by means of illuminating the array with a light source and detecting an image of the array.
  • the invention provides an apparatus for analysing a biological or biochemical assay which apparatus comprises a support structure for a test sample having an array of test locations, said support structure comprising at least one planar surface, and said apparatus further comprising an optical detector for detecting optical signals from said sample, the detector having a planar array of separate light detecting devices each corresponding to a pixel in the detector output and each having a position on the array corresponding to a respective desired test location, said planar array of light detecting devices facing said first planar surface substantially in parallel and in sufficient proximity thereto with said test locations being in between the said first planar surface and said array of light detecting devices, such that light from an array of test locations in the assay passes from the sample to a respective pixel location in the detector array without requiring an optical imaging system to focus an image of the array of test locations onto the array of light detecting devices.
  • the invention also provides an apparatus wherein said support structure comprises an output window transparent to said optical signals to be output from the sample and wherein said detecting devices are being located in a planar array
  • Said optical signals may be generated from samples on a sample chip located in an aperture in a wall of the flow cell opposite said output window and parallel thereto.
  • Said light signals are generated from samples within the cavity of the flow cell, the depth of samples within the flow cell perpendicular to said output window being controlled to avoid more than one sample being stacked on the same output path.
  • the invention also provides an apparatus for analysing a biological or biochemical assay which apparatus comprises a support structure for a test sample having an array of test locations, said support structure comprising at least one planar surface, and said apparatus further comprising an optical detector for detecting optical signals from said sample, the detector having a planar array of separate light detecting devices each corresponding to a pixel in the detector output and each having a position on the array corresponding to a respective desired test location, said planar array of light detecting devices facing said first planar surface substantially in parallel and in sufficient proximity thereto with said test locations being in between the said first planar surface and said array of light detecting devices, such that light from an array of test locations in the assay passes from the sample to a respective pixel location in the detector array without requiring an optical imaging system to focus an image of the array of test locations onto the array of light detecting devices.
  • Said support structure may comprise an output window transparent to said optical signals to be output from the sample and wherein said detecting devices are being located in a planar array in contact with said window.
  • Said support structure may comprise an output window transparent to said optical signals to be output from the sample and wherein said detecting devices are being located in a planar array facing said window in close proximity thereto.
  • Said detecting devices may be located in a planar array with a pitch separation in the array corresponding to the pitch separation of test locations in the sample and the array of detecting devices being located such that the average distance between the planar array of detecting devices and the test locations is less than 5 times said pitch separation.
  • the average distance between the planar array of detecting devices and the test locations may preferably be less than 2 times said pitch separation.
  • the average distance between the planar array of detecting devices and the test locations may preferably be less than 0.5 times said pitch separation, or even less than 0.25 times.
  • Said support may structure include a flow cell and said window may be provided by a transparent wall region of the cell.
  • a sample chip holding an array of biological or biochemical samples may be releasably secured to the flow cell adjacent an aperture in a wall of the cell opposite said output window such that said samples are exposed in the cavity of the flow cell.
  • the apparatus may comprise a stack of planar elements in contact with each other at least some of said planar elements forming the flow cell and a further planar element providing the array of optical detectors.
  • a sample chip may form one of said planar elements.
  • the invention further provides an apparatus for performing and reading out a biological or biochemical assay in a flow cell the flow cell being contained between an top surface, side walls and a bottom surface, the bottom surface comprising the top surface of a sample chip, the sample chip comprising biological or biochemical samples in an array characterized in that an array of optical detectors is directly apposed to the top surface of said flow cell to read out optical signals emitted from the samples on the sample chip such that substantially one vertex in the array of samples is imaged onto one vertex or pixel of the array of detectors.
  • Figure 1 is an exploded perspective view of a flow cell system of the present invention
  • Figure 2 is a cross section along surface A-A of the assembled flow cell
  • Figure 3 is a cross section of a lens array, which may be used in preferred embodiments of the flow cell system of the invention
  • Figure 4 is a cross section of an alternative of the lens array, which may be used in preferred embodiments of the flow cell system of the invention.
  • Figure 5 is a cross section of an array of holes in a substrate, which may be used in preferred embodiments of the flow cell system of the invention
  • a flow cell is being formed by a bottom substrate 10 with a recess 11 receiving an assay chip 30 and a micromachined lid 20.
  • the assay chip 30 has alignment features 31 which mate with alignment features 12 formed around the recess 11 of the bottom substrate 10 and carries an array of biochemical or biological samples 32 on its top surface 33.
  • the samples may for example be various oligonucleotides which are attached at predetermined locations in an array either at their 3' or at their 5' end. Alternatively the samples may be polypeptides or functional proteins that are attached at predetermined locations in the array.
  • the lid 20 is recessed by micromachining the monolithic substrate 20, e.g.
  • the lid 20, the bottom substrate 10 and the chip 30 may each be made from materials such as silica or quartz glass or, depending on the wavelength studied, silicon or silicon on insulator.
  • the recess 21 further defines inlet and outlet regions at either end of a central assay and detection region of the flow cell.
  • Inlet and outlet ports 25 and 26 are provided by vias that extend from the in and outlet regions of the lid directly through the lid to its top side. These vias can be formed by means of the same or similar micromachining techniques as the recess 21.
  • the bottom surface 21' of the recess 21 forms the top surface of the flow cell, which, in its center, has a detection region 27.
  • the detection region 27 is transparent at a wavelengths or band of wavelength to be detected by an array of detectors 41.
  • the array of detectors 41 is provided on the bottom surface 42 of a detector chip 40 which may be a CCD chip.
  • the detector chip 40 is mounted at a predetermined location directly above the detection region 27 on top of the lid 20 with the bottom surface facing downwards.
  • the lid 20 may be made of two parts, i.e. a top substrate whose bottom surface defines the surface 21' of the recess and a bottom substrate, which is bonded to the top substrate, whereby the bottom substrate is machined through to provide the side walls for the recess 21 once it is bonded to the top substrate of the lid.
  • a top substrate whose bottom surface defines the surface 21' of the recess
  • a bottom substrate which is bonded to the top substrate, whereby the bottom substrate is machined through to provide the side walls for the recess 21 once it is bonded to the top substrate of the lid.
  • Such an optional arrangement is indicated in Fig. 1 by the dashed line 29 which represents the bond line between the bottom substrate and the top substrate of the lid 20.
  • Such an arrangement has the advantage that a high optical quality may be achieved for the surface 21', e.g. by polishing, which may be required for the detection window 27.
  • the closed flow cell is shown where the lid 20 engages with the bottom substrate 10 by means of first fiduciary marks or alignment features 24 (shown in Figure 1 ) on the bottom surface 28 of the lid 20 and second fiduciary marks of alignment features 13 provided top surface 14 of the bottom substrate 10.
  • the bottom substrate 10 engages the chip 30 carrying the array of biochemical or biological samples 32 as described before.
  • the pitch between samples in the array is a distance a, which is substantially the same in magnitude as the pitch of the vertixes or pixels in the detector chip 40 so that there is a one to one correspondence between the positions of detectors in the detector array and the respective test locations in the test sample.
  • the distance between light emitting portions of the samples in the array 32 and the pixels in the array is shown as b, which is preferably similar in magnitude to the distance a.
  • flanged tubes 60 are shown, which connect via flanges 61 to the inlet and outlet ports 25, 26.
  • a fluid tight connection may be achieved by means of a press fit, a permanent connection, e.g. by means of glue, resin or solder or similar means well known in the art.
  • a wafer fabrication method is used for making the lid 20 and the bottom substrate 10
  • direct wafer-to-wafer bonding may be used to bond the lid 20 to the bottom substrate 10.
  • Other bonding techniques may be used, e.g. by means of a UV cured resin or glue.
  • the engagement between the bottom substrate 10 and the assay chip 30 is non-permanent, but the alignment features are made such that a fluid tight press seal can be made between the bottom substrate and the chip when they engage with each other.
  • the array of biological or biochemical samples 32 is aligned in parallel with the array of detectors 41 with each vertex in the sample array 32 being directly opposite one vertex or pixel in the array of detectors 41.
  • the flow cell may have a temperature control means (not shown), e.g. comprising a thermistor and a Peltier device to enable control of the flow cell and its contents as desired during operation.
  • a temperature control means e.g. comprising a thermistor and a Peltier device to enable control of the flow cell and its contents as desired during operation.
  • the chip 30 has a planar surface and is made of a material that is transparent at a first wavelength emitted by a light source 50.
  • the one or both of the top and bottom surfaces of the chip may be polished to provide for a sufficiently high optical quality of these surfaces.
  • the light source 50 emits light at the first wavelength or band of wavelengths.
  • appropriate materials for forming the chip are for example, silica, quartz glass, silicon or silicon on insulator.
  • the light is collimated by a lens 51 onto the chip 30 and is transmitted through the chip on to the array of biochemical or biological samples 32 located on the top surface of the chip 33.
  • the fluorescent marker may be for example R-phycoerythrin (R- PE), allophycocyanin (APC), flourescinisothiocyanate (FITC), rhodamine, or texas red.
  • R- PE R-phycoerythrin
  • APC allophycocyanin
  • FITC flourescinisothiocyanate
  • rhodamine rhodamine
  • fluorescent light emitted from markers is detected by the vertex or pixel in the CCD or other detector chip that is directly apposed to this vertex and the detection of fluorescence is possible at all vertexes in the array at the same time.
  • the array of detectors 41 is directly apposed to the top surface of the flow cell 21 and its vertexes or pixels are matched with those in the sample array substantially one to one, this provides for a very compact arrangement for a flow cell with high resolution and a view field that is not limited by the resolution required.
  • an external light source 50 is used to cause fluorescence of the samples by putting the light source on the opposite side of the chip 30 from the detector 40 and illuminating the sample array 32 through the chip 30 allows easier management of components in a workstation and helps to reduce the size of the entire device.
  • the height of the flow cell or the distance between the marker molecules in the sample array and the pixels in the array of detectors is similar in magnitude to the spacing between vertices in the array.
  • the detector 40 In order to improve the detection of a fluorescent signal at a second wavelength in response to a primary excitation of light a first wavelength it is possible to provide the detector 40 with a bandpass filter by means of an optical coating deposited on the bottom surface 42 of the detector chip 40 or on the detection window 27 of the lid 20.
  • Such optical coatings are well known in the art and can, for example, be formed by a multi-layer of dielectric films.
  • the band pass filter substantially reflects light at the first wavelength and passes light at the second wavelength.
  • the detector 40 may be such that it is more responsive at the second wavelength than the first wavelength.
  • the detector material could be transparent at the first wavelength and a high absorption region for the first wavelength, i.e.
  • the assay chip 30 itself may be provided with a dielectric coating that specifically reflects light at the second wavelength or band of wavelengths, but that is transparent at the first wavelength or band of wavelengths. If such an arrangement is used, fluorescent light emitted by the • marker molecules away from the detector can also be captured by the detector chip 40. If the light source is sensitive to back reflections, an optical isolator or circulator can be placed between the light source and the array chip.
  • the array chip 30 can be designed to fulfill both functions.
  • the array chip 30 is made large enough to cover the substantially the whole perimeter of the lid.
  • fiduciary marks are employed to ensure that the chip will register with the lid 20 at a predetermined location, such that the array on the chip is aligned with the array of pixels in the detector.
  • it is more advantageous to provide the chip 30 and bottom substrate 10 as separate parts or one part depends on the design of the device and whether it is more important to save area of chip substrate in high volume applications, or simplify the design of the system.
  • FIG. 3 and Fig. 4 further improved embodiments of the present invention are shown.
  • an array of lenses 70 is formed in a substrate 80, whereby the lens array is matched to the array of biological samples 32 and the array of pixels 41 in the detector chip 40 such that samples at a each vertex of the sample array are imaged on the corresponding vertex or pixel in the detector array by one corresponding lens in the lens array.
  • Such an arrangement can provide a better spatial resolution as well as lower cross talk between adjacent vertexes in the sample array.
  • the lens array can be formed in a wafer-based substrate, e.g. silicon or silica.
  • a mask of the array is produced on the substrate using known photolithography techniques and an isotropic etch is used to form spherical recesses in the substrate.
  • the recesses are then filled by depositing a film of a second material.
  • the substrate material is glass
  • the second material can be a glass with higher refractive index than the glass of the substrate material. Techniques for varying the refractive index of glass are well known by means of doping.
  • the second material may also be glass, but in this case a selective etch is performed from the side of the substrate opposite that from which the first etch has been performed to form a recess at the location of each lens that takes away the silicon around the center of the lens and that stops at the curved glass structure of the second material deposited previously form the other side.
  • the substrate is polished back to a planar surface on the side where the recesses were filled.
  • the regions between lenses of the array can be doped separately to provide for additional optical isolation in the substrate between the lens regions. This will generally reduce the amount of light scattered between lenses and improve crosstalk in the system.
  • an array of holes or vias may be formed in a substrate that is disposed between the array of sample and the array of the detectors.
  • the substrate in which the holes or vias are formed is preferably opaque at the wavelength range to be detected.
  • the crosstalk between adjacent vertexes in the array is thereby limited by the maximum angle at which light can pass through one of the holes to reach a given pixel on the detector.
  • the chip carrying the sample array may directly abut against the bottom surface of the hole mask or the lens array to improve the optical cross-talk isolation between different sample locations of the device.
  • the material of the hole mask or the lens array in between the holes or lenses, respectively is opaque, it is possible to achieve almost 100% cross-talk isolation without having to provide micromachined features on the substrate of the chip itself. This would be advantageous if the chip itself is to be a low-cost disposable device.
  • the array may be located between the array chip 30 and the recess 21 of the flow cell or alternatively between the top surface of the flow cell and the detector chip 40.
  • the system of the present invention operates on the basis of in-line attenuation, rather than fluorescence.
  • the binding reaction may be observed merely by the attenuation caused by a specific binding reaction on a vertex of the sample array, rather than the detection of fluorescently labeled ligands when light emitted from the light source onto the samples is attenuated when passing the samples on its way to the array of detectors.
  • the maximum of the responsiveness of the pixels in the detector is at the first wavelength or band of wavelengths.
  • no primary light source is required and the biological or biochemical assay performed on the sample array is carried out on the basis of chemiluminescence or electrochemiluminescence of a tagged ligand.
  • chemiluminescence or electrochemiluminescence of a tagged ligand For example it would be possible to provide ligands with tagged luciferase or a similar chemiluminescent protein that will start to emit light when the flow cell is filled with a solution containing ATP.
  • electrochemiluminescent ligands e.g. ones that contain ruthenium metal ions
  • the assay chip is formed from a conducting substrate such as a doped semiconductor or a metal.
  • An isolation layer may be provided between the assay chip and the lid isolating the assay chip from the lid electrically.
  • the lid itself has conducting parts, such that a voltage can be applied between the conducting parts of the lid and the assay chip.
  • the chip which already carries samples that have been immobilized in an array on its top surface, is engaged and clamped to the open lid of the flow cell such that the flow cell is now closed.
  • the samples on the chip may already have undergone one or more pre-processing steps prior to the chip being inserted into the flow cell.
  • This step may be performed by an automated cassette-to- cassette chip handling system for high throughput applications.
  • the flow cell is then washed with an appropriate wash buffer to remove any impurities and prime the flow cell. Subsequently the array is stained or hybridized with a buffer containing the staining or hybridization reagent.
  • the flow cell is washed again either with the same or a different wash buffer to remove excess staining or hybridization reagent.
  • the measurement is carried out employing one of the methods outlined above, i.e. by using any of fluorescence, absorption, chemiluminescence, or electrochemi-luminescene.
  • a new staining experiment can be performed on the same chip or a new chip can be used. If a new chip is used, the old one may be removed and replaced with a new chip by the automated handling system, preceded by a wash or purge step, clearing the flow cell of liquid.
  • the flow cell may be washed first with a release buffer, releasing the staining reagent or hybridization reagent from its targets, followed by a further wash step.
  • temperature variations may be used in conjunction with wash steps to remove the staining or hybridization reagents.
  • Very small cell volumes can be achieved by using the micromachining methods described above which can accelerate fluid handling during operation of the flow cell. Furthermore the present invention allows screening of very large arrays, e.g. of millions of samples on a chip of one square inch. The invention further allows easy combination of the flow cell with automated chip handling systems, such as cassette-to-cassette handling. When wafer based micromachining techniques are used with the present invention, as described above, it is further possible to replace several seals normally requiring several sealing materials with mere pressure seals, thus reducing the complexity of the device further.
  • the apparatus described may also be used to read out a signal from samples in an array that are immobilized on a chip which have been pre-stained or pre-processed, such that no further liquid handling is required before a readout operation takes place. It will be appreciated that in such a situation certain simplifications can be made in the design of the apparatus according to the invention.
  • the flow cell system described can be used to count cell populations that bind to a specific marker, e.g. a phenotypic marker.
  • a specific marker e.g. a phenotypic marker.
  • the bottom surface of the flow cell does not have to comprise a sample chip carrying a biological or biochemical sample.
  • the bottom substrate may just be a transparent or opaque substrate, depending on whether a fluorescent, absorptive or luminescent method for detection is used. If the detection method is by fluorescence the following protocol may be carried out:
  • a suspension of cells of interest is stained using a staining reagent that is specific to the phenotypic marker of interest. For example, if one wishes to detect the percentage of CD8+ T cells in a sample of peripheral blood mononuclear cells (PBMCs) the cells may be labeled with a fluorescent tagged anti CD8 monochlonal antibody using methods well known in the art. Either the whole cell suspension sample or a fraction thereof is subsequently moved into the flow cell. The amount of cells in the flow cell at any one time is thereby controlled to be substantially less than the number of pixels or vertexes in the detector array, e.g. for a detector array with 3x10E6 pixels the number of cells in the cell may be 3x10E5.
  • PBMCs peripheral blood mononuclear cells
  • a measurement is taken by shining light of a first wavelength through the transparent bottom substrate onto the cell sample, which causes the fluorescent markers to fluoresce at a second wavelength.
  • Labeled cells will appear as isolated dots on the array of pixels in the detector and the number of fluorescent dots can simply be counted out by electronic means.
  • the detector array can be designed as a stacked array in which two detection regions are stacked vertically. The first of these two regions nearer to the sample may be responsive to the second wavelength of fluorescent light and transparent to the first wavelength of illuminating light. The second region is responsive only to the first wavelength of illuminating light. This may be achieved by engineering the optical band gaps of the materials from which the detection regions are formed using methods well known in the art.. With this detector design it is possible to detect labeled cells by means of fluorescence and unlabeled cells by means of in-line absorption at the same time. The ratio of labeled and unlabeled cells can hence be determined in a straightforward manner.
  • an external magnetic field e.g. generated by a coil that is wound around the cell, can then be used to immobilize the cells on the bottom or top surface of the flow cell. Once the measurement has been taken, the field can be switched off to allow the flow cell to be cleared of the cell sample by washing.
  • the height of flow cell may be selected such that, on average, cells in the flow cell will not overlap in their horizontal position by being stacked vertically.
  • the array of detectors is located sufficiently close to the array of test locations that no optical imaging system is needed to focus one image of the array of test locations, or even one image of a group of test locations, onto the array of light detecting devices.
  • Each detector in the detector array is arranged to detect light from a single respective test location. The detection by the detector array is carried out simultaneously for a plurality of test locations until minimum cross talk between the signals from the separate test locations.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Clinical Laboratory Science (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

Cette invention se rapporte à un appareil qui sert à analyser un dosage biologique ou biochimique et qui comprend à cet effet une structure de support (30) pour un échantillon de test ayant une matrice de points de test (32), cette structure de support comprenant au moins une surface plane (30) et cet appareil comprenant en outre un détecteur optique (40) destiné à détecter les signaux optiques provenant dudit échantillon. Ledit détecteur comporte une matrice plane de dispositifs de photodétection séparés (41) correspondant chacun à un pixel dans la sortie du détecteur et présentant chacun une position sur la matrice qui correspond à un point de test souhaité correspondant. Cette matrice plane de dispositifs de photodétection est située face à la première surface plane (42) dans une position essentiellement parallèle et à proximité suffisante par rapport à elle, lesdits points de test étant situés entre la première surface plane et la matrice des dispositifs de photodétection, pour que la lumière provenant d'une matrice de points de test passe de l'échantillon à un point de pixel correspondant dans la matrice des détecteurs, sans qu'il soit nécessaire de recourir à un système d'imagerie optique pour la mise au point d'une image de la matrice des points de test sur la matrice des dispositifs de photodétection.
PCT/GB2001/004933 2000-11-08 2001-11-07 Analyse de dosages biologiques et biochimiques WO2002039096A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002212520A AU2002212520A1 (en) 2000-11-08 2001-11-07 Analysis of biological and biochemical assays

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0027293A GB2368903A (en) 2000-11-08 2000-11-08 Analysis of biological and biochemical assays
GB0027293.0 2000-11-08

Publications (1)

Publication Number Publication Date
WO2002039096A1 true WO2002039096A1 (fr) 2002-05-16

Family

ID=9902797

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2001/004933 WO2002039096A1 (fr) 2000-11-08 2001-11-07 Analyse de dosages biologiques et biochimiques

Country Status (3)

Country Link
AU (1) AU2002212520A1 (fr)
GB (1) GB2368903A (fr)
WO (1) WO2002039096A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004015488A1 (de) * 2004-03-26 2005-10-20 Lavision Biotec Gmbh Verfahren zum Auslesen durch streifenweises Abtasten von flächigen Objekten mit Substanzen, die Fluoreszenzstrahlung abgeben
WO2014201520A1 (fr) * 2013-06-19 2014-12-24 Ellume Pty Ltd Dispositif d'analyse utilisant des étiquettes fluorescentes
CN109085157A (zh) * 2017-06-14 2018-12-25 淳洁 用于化学发光检测的探测单元、电子暗盒压片装置及其用途

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050239195A1 (en) * 2002-06-13 2005-10-27 Millenium Biologix Ag Reaction chamber
CN103473777B (zh) * 2013-09-18 2016-12-07 陕西中莱节能有限公司 一种基于数字图像的led芯片插深和插浅缺陷的检测算法
US11708551B2 (en) * 2017-10-06 2023-07-25 Wyatt Technology Corporation Temperature uniformity and suppressing well plate warping in high throughput measurements

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028538A1 (fr) * 1995-03-10 1996-09-19 Meso Scale Technologies, Llc Essai par electrochimioluminescence multispecifique et multirangee
WO1997045730A1 (fr) * 1996-05-30 1997-12-04 Biodx Procedes et appareil a groupement miniaturise de cellules pour le criblage sur la base de cellules
WO1998026277A2 (fr) * 1996-12-12 1998-06-18 Prolume, Ltd. Appareil permettant de detecter et d'identifier des agents infectieux et procede correspondant
WO2000012759A1 (fr) * 1998-08-26 2000-03-09 Alpha Innotech Corporation Systeme de detection sur biopuce
WO2000012123A2 (fr) * 1998-08-28 2000-03-09 Febit Ferrarius Biotechnology Gmbh Procede et dispositif de mesure pour la determination d'une pluralite d'analytes dans un echantillon

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2014300A (en) * 1977-07-29 1979-08-22 Opto Electronic Displays Ltd Opacity measurement apparatus and method
US5073029A (en) * 1990-02-16 1991-12-17 Eqm Research, Inc. Multisource device for photometric analysis and associated chromogens
US5736410A (en) * 1992-09-14 1998-04-07 Sri International Up-converting reporters for biological and other assays using laser excitation techniques
AT403745B (de) * 1996-02-29 1998-05-25 Avl Verbrennungskraft Messtech Messanordnung mit einem für anregungs- und messstrahlung transparentem trägerelement
GB2315131B (en) * 1996-07-10 2000-11-29 Cambridge Imaging Ltd Improvements in and relating to imaging
AU4607197A (en) * 1996-09-26 1998-04-17 Sarnoff Corporation Massively parallel detection
US6200531B1 (en) * 1998-05-11 2001-03-13 Igen International, Inc. Apparatus for carrying out electrochemiluminescence test measurements
EP1190232A1 (fr) * 1999-06-26 2002-03-27 Packard Instrument Company, Inc. Lecteur pour microplaques

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028538A1 (fr) * 1995-03-10 1996-09-19 Meso Scale Technologies, Llc Essai par electrochimioluminescence multispecifique et multirangee
WO1997045730A1 (fr) * 1996-05-30 1997-12-04 Biodx Procedes et appareil a groupement miniaturise de cellules pour le criblage sur la base de cellules
WO1998026277A2 (fr) * 1996-12-12 1998-06-18 Prolume, Ltd. Appareil permettant de detecter et d'identifier des agents infectieux et procede correspondant
WO2000012759A1 (fr) * 1998-08-26 2000-03-09 Alpha Innotech Corporation Systeme de detection sur biopuce
WO2000012123A2 (fr) * 1998-08-28 2000-03-09 Febit Ferrarius Biotechnology Gmbh Procede et dispositif de mesure pour la determination d'une pluralite d'analytes dans un echantillon

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004015488A1 (de) * 2004-03-26 2005-10-20 Lavision Biotec Gmbh Verfahren zum Auslesen durch streifenweises Abtasten von flächigen Objekten mit Substanzen, die Fluoreszenzstrahlung abgeben
US7211807B2 (en) 2004-03-26 2007-05-01 Lavision Biotec Gmbh Readout method performed by stripe scanning planar objects with substances emitting fluorescence radiation
DE102004015488B4 (de) * 2004-03-26 2008-01-31 Lavision Biotec Gmbh Verfahren zum Auslesen durch streifenweises Abtasten von flächigen Objekten mit Substanzen, die Fluoreszenzstrahlung abgeben
WO2014201520A1 (fr) * 2013-06-19 2014-12-24 Ellume Pty Ltd Dispositif d'analyse utilisant des étiquettes fluorescentes
US10578556B2 (en) 2013-06-19 2020-03-03 Ellume Limited Assay device employing fluorescent labels
US11442012B2 (en) 2013-06-19 2022-09-13 Ellume Limited Assay device employing fluorescent labels
CN109085157A (zh) * 2017-06-14 2018-12-25 淳洁 用于化学发光检测的探测单元、电子暗盒压片装置及其用途

Also Published As

Publication number Publication date
GB0027293D0 (en) 2000-12-27
GB2368903A (en) 2002-05-15
AU2002212520A1 (en) 2002-05-21

Similar Documents

Publication Publication Date Title
JP6573864B2 (ja) 分子または粒子の超高感度検出用のシステム、デバイスおよび方法
US6437345B1 (en) Sensing unit provided with separated detection light guiding
US7799558B1 (en) Ligand binding assays on microarrays in closed multiwell plates
US11112338B2 (en) Dual-image based bioimaging devices and techniques
US7879598B2 (en) Multianalyte determination system and methods
EP1983331B1 (fr) Système optique de lecture de biopuce
US6795192B2 (en) SPR sensor and SPR sensor array
EP2291643B1 (fr) Système et procédé de caractérisation de microréseau
CA2304059C (fr) Dispositif de realisation d'essais photometriques
US9557259B2 (en) Optical particle detector and detection method
US8975607B2 (en) Confocal optical detector, detector array, and manufacturing method thereof
US20060105395A1 (en) Methods and systems for positioning microspheres for imaging
JP2009204616A (ja) 共鳴光散乱粒子標識から発生する信号を読み取るための装置
EP1938049B1 (fr) Capteur de luminescence comprenant au moins deux grilles de fil
WO2008155716A1 (fr) Dispositif de détecteur microélectronique pour détecter des particules de marqueur
US9863863B2 (en) Apparatus for cluster detection
US7863037B1 (en) Ligand binding assays on microarrays in closed multiwell plates
WO2002039096A1 (fr) Analyse de dosages biologiques et biochimiques
EP1190235B1 (fr) Analyse par imagerie pour microechantillons
US20090041633A1 (en) Apparatus and method for performing ligand binding assays on microarrays in multiwell plates
WO2008103200A2 (fr) Dispositif et procédé pour effectuer des essais de liaison de ligands sur microréseaux de plaques à plusieurs puits
KR102551560B1 (ko) 재귀반사 신호 측정용 가젯
US20070231881A1 (en) Biomolecular interaction analyzer
WO2011102981A1 (fr) Imagerie haute résolution ne nécessitant pas l'utilisation de marqueurs
WO2002066965A2 (fr) Appareil de dosage, procede de dosage et ensemble de sondes utilise

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP