WO2001007895A1 - Biochip reading device - Google Patents

Biochip reading device Download PDF

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Publication number
WO2001007895A1
WO2001007895A1 PCT/FR2000/002143 FR0002143W WO0107895A1 WO 2001007895 A1 WO2001007895 A1 WO 2001007895A1 FR 0002143 W FR0002143 W FR 0002143W WO 0107895 A1 WO0107895 A1 WO 0107895A1
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WO
WIPO (PCT)
Prior art keywords
chip
information
light
scanning
pad
Prior art date
Application number
PCT/FR2000/002143
Other languages
French (fr)
Inventor
Philippe Peltie
Raymond Campagnolo
Patrick Chaton
Original Assignee
Commissariat A L'energie Atomique
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Publication of WO2001007895A1 publication Critical patent/WO2001007895A1/en

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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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • 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
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00158—Elements containing microarrays, i.e. "biochip"

Definitions

  • the invention relates to a biological chip reading device.
  • the invention relates to a device for reading a nucleic acid chip commonly called a DNA chip (DNA for deoxyribonucleic acid).
  • a DNA chip DNA for deoxyribonucleic acid.
  • the invention finds applications in numerous fields such as, for example, the field of molecular biology for the recognition of antibodies and antigens, the sequencing of genomes, the search for mutations responsible for diseases, bacterial detection, etc.
  • Biological chip reading devices use the fluorescence technique.
  • the fluorescence technique consists in mixing a substance whose identification and / or concentration is to be determined with a substance having the capacity to emit fluorescent light under the action of radiation.
  • a DNA chip consists of a matrix of
  • N x M pads each pad containing a mixture of substances as mentioned above, that is to say capable of emitting fluorescent light by illumination.
  • a DNA chip reading device includes means for illuminating the chip and means for detecting fluorescent light from the chip.
  • the lighting means of the chip generally comprises a laser source, optical means for shaping the beam coming from the laser source, means making it possible to direct the laser beam towards the chip and means making it possible to scan the laser beam at the surface of the chip.
  • the means for detecting the fluorescent light coming from the chip generally comprises optical means for shaping the fluorescent light, means for detecting the fluorescent light and electronic means for processing the signals coming from the detection means.
  • the photodetectors are made from silicon material in MOS technology (MOS for "Metal Oxide Semiconductor”).
  • MOS Metal Oxide Semiconductor
  • the laser source and the scanning means use other technologies and materials other than silicon. It is then necessary to carry out a transfer from the laser source / scanning part to the photodetector part. This postponement is a delicate and therefore costly operation.
  • the invention does not have this drawback.
  • the invention relates to a device for reading an active biological chip comprising at least one line of M pads, M being an integer greater than or equal to 1, the device comprising an optical unit consisting of means for illuminating the chip comprising a laser source and scanning means for scanning, on the surface of the chip, a laser beam coming from the laser source.
  • the device comprises means for constituting information (I) relating to the position of at least one stud of the chip with respect to the lighting means and means for constituting a control signal (B) of the scanning means from of said information (I).
  • the invention also relates to an active biological chip comprising at least one line of M pads (P), M being an integer greater than or equal to 1. Each pad is surrounded by a reflecting zone (Z).
  • a biological chip reading device is an easily transportable miniaturized system, usable both outside the laboratory and in the laboratory and allowing excellent performance to be obtained.
  • the means for illuminating the chip comprise a laser source, optical means for shaping a light beam coming from the laser source, means for directing the laser beam towards the chip and means for scanning the laser beam on the surface of the chip.
  • the printed circuit comprises electronic circuits for supplying and controlling, inter alia, the laser source, the photodetector means, the scanning means, as well as a memory area for storing the data coming from the fluorescence detection means.
  • the biological chip is an active chip.
  • active chip is meant a chip on which are integrated the photodetectors which receive the fluorescence emission.
  • the active chip can also contain computer memories for recording information.
  • the field scanned by the light beam is larger than the area of the chip. It is then not necessary to move the chip relative to the lighting means. In order for the chip to be scanned correctly, it is therefore sufficient to mark the studs.
  • the active chip is disposable.
  • the reading system then comprises, on the one hand, a reading head which is not disposable consisting of lighting means and scanning means and, on the other hand, disposable photodetection means integrated into the active chip .
  • FIGS. 1A and 1B represent a device for reading a biological chip according to the invention
  • FIG. 2 shows a biological chip pad according to a particular embodiment of
  • Figures 1A and IB show a biological chip reading device according to the invention.
  • FIG. 1A represents a perspective view of the device.
  • the reading device comprises a printed circuit 1 and an optical unit 2.
  • the printed circuit 1 is provided with a connector 5 into which is plugged, during the reading of the chip, a support 3 on which the chip 4 is positioned.
  • the optical unit 2 comprises means for illuminating the chip and means for detecting the reflected and / or scattered light coming from the chip.
  • the optical unit 2 is described in more detail in FIG. 1B.
  • the optical unit 2 is plugged into the printed circuit 1 and the support 3 in the connector 5.
  • the precision of the positioning of the optical unit in the printed circuit as well as the precision of the positioning of the support 3 in the connector 5 be greater than 1/10 th of a millimeter.
  • the reading device according to the invention advantageously has a very small footprint as well as great maneuverability.
  • the size of such a system can be, for example, of the order of 200 ⁇ 100 ⁇ 100 mm 3 .
  • FIG. 1B represents a detailed view of the optical unit 2 in connection with a biological chip 4.
  • the optical unit 2 comprises a lighting means 6 comprising a laser source, means 7 for scanning, in two dimensions, of a beam coming from the laser source, means 8 for shaping the beam coming from the scanning means 7, means 9, 10 for directing and focusing the laser beam towards the chip 4, a detection optic 11 and photodetector means 12 for receiving and detecting the light returned by the chip 4.
  • a lighting means 6 comprising a laser source, means 7 for scanning, in two dimensions, of a beam coming from the laser source, means 8 for shaping the beam coming from the scanning means 7, means 9, 10 for directing and focusing the laser beam towards the chip 4, a detection optic 11 and photodetector means 12 for receiving and detecting the light returned by the chip 4.
  • the scanning means 7 consist of an acousto-optical deflector.
  • the scanning of the chip allows the studs to be illuminated one after the other.
  • the laser source does not therefore need to emit a high power signal.
  • the illumination of a 50 ⁇ m side pad can be carried out under excellent conditions with a signal of 0.25 mW of power. Scanning the chip also improves the signal-to-noise ratio.
  • the laser beam contained in the lighting means 6 may have a power substantially less than 5 mW and a diameter of the order of 150 ⁇ m.
  • the light signal returned by the chip 4 contains information relating to the position of the pads relative to the lighting means.
  • the photodetector means 12 comprise means for converting the light information relating to the position of the pads into electrical information I transmitted to a processing circuit 13.
  • the signal S from the processing circuit 13 is transmitted to a circuit 14 for controlling the scanning means 7 which also receives a setpoint signal C.
  • the circuit 14 for controlling the scanning means 7 delivers a scanning control signal B from the error signal resulting from the comparison of the signal S and the reference signal C. It is then possible to obtain an alignment of the pads with very precise lighting means. An accuracy of less than 5 ⁇ m can thus be obtained for studs with a side of 50 ⁇ m.
  • the processing circuit 13 and the circuit 14 for controlling the scanning means 7 are preferably produced on the printed circuit 1.
  • the lighting means 6 comprise, in addition to the laser source, at least one diode light-emitting to illuminate the chip 4 in whole or in part.
  • the detection optic 11 then comprises a matrix of microlenses and the detection means 12 comprise a CCD matrix (the acronym CCD from the English “Charge Coupled Device”).
  • the image formed by the CCD matrix due to the illumination of the chip by the light emitted by the light-emitting diode exhibits a moire effect in the case where the frame defined by the microlenses is offset with respect to the frame defined by the pads of the chip.
  • the electrical signal which translates the moiré effect constitutes, according to the first embodiment of the invention, the information I which represents the misalignment between the chip 4 and the lighting means 6.
  • the circuit 13 for processing the electrical information I makes it possible to store the image of the chip 4 and to locate the position of the pads of the chip in an absolute reference.
  • the setpoint signal C is then representative of the position that the pads must occupy to ensure ideal alignment between the lighting means of the chip and the chip.
  • it is the scattered light coming from the biological chip which is used to align the pads with the lighting means.
  • the detection means 12 then comprise a photodiode for detecting the amplitude of the light scattered by a pad of the chip.
  • the correct position a pad relative to the lighting means is then detected by a maximum of the light scattered at the excitation wavelength.
  • Information I then consists of the detected electrical signal of maximum amplitude.
  • This alignment technique is particularly advantageous when the studs of the chips are relatively spaced, as is the case, for example, for MICAM® chips.
  • each pad P of a biological chip is surrounded by a reflecting zone Z.
  • the zone Z is preferably of circular shape as shown in FIG. 2.
  • the zone Z is metallized so as to reflect the light it receives.
  • the detection means 12 then comprise a four-quadrant detector on which the image of the zone Z is formed. After the search for a reference pad and taking into account the positioning error of the chip relative to this reference pad, the scanning means 7 are programmed to bring the lighting of the chip opposite a plot P so that an image of the plot is formed on the four-quadrant detector.
  • the four-quadrant detector then makes it possible to create two positioning error signals, a first error signal in a first direction and a second error signal in a second direction perpendicular to the first direction.
  • the two error signals constitute the information I transmitted to the processing circuit 13.
  • This third embodiment of the invention has the advantage of maintaining the enslavement of the lighting means on a pad even though a mechanical disturbance may occur at the level of the reader. Furthermore, it is not necessary that the movement of the light beam from one stud to another takes place in a precise manner.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Optics & Photonics (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

The invention concerns an active biological chip reading device comprising an optical block (2) to illuminate the chip (4) and means for capturing (11) and detecting (12) light reflected by the chip. The optical block illuminating the chip comprises laser beam scanning means (7) at the surface of the chip. The reading device comprises means to constitute an information (I) concerning the position of at least one pad of the chip relative to the illuminating means and to constitute, from said information (I), a signal controlling (B) the scanning means (7). The invention is applicable in the field of molecular biology for identifying antibodies and antigens.

Description

DIPOSΓΠF DE LECTURE DE BIOPUCE BIOPUCE READING DIPOSΓΠF
Domaine technique et art antérieurTechnical field and prior art
L'invention concerne un dispositif de lecture de puce biologique.The invention relates to a biological chip reading device.
Plus particulièrement, l'invention concerne un dispositif de lecture de puce à acides nucléiques communément appelée puce à ADN (ADN pour Acide désoxyribonucléique ) . L'invention trouve des applications dans de nombreux domaines tels que, par exemple, le domaine de la biologie moléculaire pour la reconnaissance des anticorps et antigènes, le séquençage des génomes, la recherche de mutations responsables de maladies, la détection bactérienne, etc.More particularly, the invention relates to a device for reading a nucleic acid chip commonly called a DNA chip (DNA for deoxyribonucleic acid). The invention finds applications in numerous fields such as, for example, the field of molecular biology for the recognition of antibodies and antigens, the sequencing of genomes, the search for mutations responsible for diseases, bacterial detection, etc.
Les dispositifs de lecture de puce biologique utilisent la technique de fluorescence.Biological chip reading devices use the fluorescence technique.
La technique de fluorescence consiste à mélanger une substance dont l'identification et/ou la concentration est à déterminer avec une substance ayant la capacité d'émettre une lumière fluorescente sous l'action d'un rayonnement.The fluorescence technique consists in mixing a substance whose identification and / or concentration is to be determined with a substance having the capacity to emit fluorescent light under the action of radiation.
Une puce à ADN est constituée d'une matrice deA DNA chip consists of a matrix of
N x M plots, chaque plot contenant un mélange de substances tel que mentionné ci-dessus, c'est-à-dire susceptible d'émettre une lumière fluorescente par illumination.N x M pads, each pad containing a mixture of substances as mentioned above, that is to say capable of emitting fluorescent light by illumination.
Un dispositif de lecture de puce à ADN comprend un moyen d'éclairage de la puce et un moyen pour détecter la lumière fluorescente issue de la puce. Le moyen d'éclairage de la puce comprend généralement une source laser, des moyens optiques pour mettre en forme le faisceau issu de la source laser, des moyens permettant de diriger le faisceau laser vers la puce et des moyens permettant de balayer le faisceau laser à la surface de la puce.A DNA chip reading device includes means for illuminating the chip and means for detecting fluorescent light from the chip. The lighting means of the chip generally comprises a laser source, optical means for shaping the beam coming from the laser source, means making it possible to direct the laser beam towards the chip and means making it possible to scan the laser beam at the surface of the chip.
De même, le moyen pour détecter la lumière fluorescente issue de la puce comprend généralement des moyens optiques pour mettre en forme la lumière fluorescente, des moyens de détection de la lumière fluorescente et des moyens électroniques pour traiter les signaux issus des moyens de détection.Likewise, the means for detecting the fluorescent light coming from the chip generally comprises optical means for shaping the fluorescent light, means for detecting the fluorescent light and electronic means for processing the signals coming from the detection means.
Afin de pouvoir lire des puces à ADN hors du laboratoire (par exemple, pour le diagnostic médical ou encore pour la détection bactérienne sur site dans le domaine agro-alimentaire), il a été conçu des dispositifs intégrés utilisant des instruments moins encombrants et moins coûteux que ceux utilisés en laboratoire. Le document intitulé " Development of a DNA biochip for gène diagnosls ", T. Vo-Dlnh et al . , SPIE, vol . 3253 divulgue un microdispositif intégrant un système d'excitation/détection électro-optique et des circuits électroniques de traitement. Un tel dispositif est dépourvu de moyens de balayage de la puce.In order to be able to read DNA chips outside the laboratory (for example, for medical diagnosis or even for on-site bacterial detection in the food industry), integrated devices have been designed using less bulky and less expensive instruments than those used in the laboratory. The document entitled "Development of a DNA biochip for gene diagnosls", T. Vo-Dlnh et al. , SPIE, vol. 3253 discloses a microdevice integrating an electro-optical excitation / detection system and electronic processing circuits. Such a device does not have means for scanning the chip.
Ceci présente l'inconvénient de nécessiter un éclairage de la puce avec une source de puissance élevée (typiquement 400 W pour une puce de 2 mm x 2 mm) Le document intitulé "A review of mlcrofabricated devices for gene-based diagnostics " , M. Eggers et D. Ehrlich, Hématologie Pathology, 9 (1 ) , 1-15 ( 1995 ) divulgue un microdispositif d'analyse par electrophorese comprenant une partie microfluidique incorporé dans la puce. Le document intitulé "A microsensor array for biochemical sensing" , F. Van Steenkiste et al . , Sensors and Actuators 13, 44 ( 1997) , 409-412 divulgue un dispositif de mesure des propriétés biochimiques des liquides biologiques. De façon générale, les solutions connues pour réaliser des dispositifs de lecture miniaturisés de puces biologiques proposent soit des dispositifs présentant une structure classique - mais miniaturisée - comprenant, entre autres, un microscope, soit des dispositifs intégrés où toutes les fonctions sont regroupées .This has the drawback of requiring lighting of the chip with a high power source (typically 400 W for a 2 mm x 2 mm chip). The document entitled "A review of mlcrofabricated devices for gene-based diagnostics", M. Eggers and D. Ehrlich, Hématologie Pathology, 9 (1), 1-15 (1995) discloses a microdevice for analysis by electrophoresis comprising a microfluidic part incorporated in the chip. The document entitled "A microsensor array for biochemical sensing", F. Van Steenkiste et al. , Sensors and Actuators 13, 44 (1997), 409-412 discloses a device for measuring the biochemical properties of biological fluids. In general, the known solutions for producing miniaturized devices for reading biological chips offer either devices having a conventional structure - but miniaturized - comprising, inter alia, a microscope, or integrated devices where all the functions are grouped together.
De telles solutions présentent de nombreux inconvénients. En particulier, les dispositifs de type intégré qui regroupent l'excitation laser et le balayage augmentent très sensiblement la complexité de la puce biologique et, partant, le coût de celle-ci.Such solutions have many drawbacks. In particular, integrated type devices which combine laser excitation and scanning very significantly increase the complexity of the biological chip and, consequently, the cost thereof.
En effet, de façon générale, les photodétecteurs sont réalisés à base de matériau silicium en technologie MOS (MOS pour "Métal Oxyde Semi-conducteur"). Par ailleurs, la source laser et les moyens de balayage utilisent d'autres technologies et des matériaux différents du silicium. Il est alors nécessaire d'effectuer un report de la partie source laser/balayage sur la partie photodétecteur. Ce report est une opération délicate et, partant, coûteuse. Exposé de 1 ' inventionIndeed, in general, the photodetectors are made from silicon material in MOS technology (MOS for "Metal Oxide Semiconductor"). Furthermore, the laser source and the scanning means use other technologies and materials other than silicon. It is then necessary to carry out a transfer from the laser source / scanning part to the photodetector part. This postponement is a delicate and therefore costly operation. Statement of the invention
L'invention ne présente pas cet inconvénient. En effet, l'invention concerne un dispositif de lecture de puce biologique active comprenant au moins une ligne de M plots, M étant un nombre entier supérieur ou égal à 1, le dispositif comprenant un bloc optique constitué de moyens d'éclairage de la puce comprenant une source laser et des moyens de balayage pour balayer, à la surface de la puce, un faisceau laser issu de la source laser. Le dispositif comprend des moyens pour constituer une information (I) relative à la position d'au moins un plot de la puce par rapport aux moyens d'éclairage et des moyens pour constituer un signal de commande (B) des moyens de balayage à partir de ladite information (I).The invention does not have this drawback. In fact, the invention relates to a device for reading an active biological chip comprising at least one line of M pads, M being an integer greater than or equal to 1, the device comprising an optical unit consisting of means for illuminating the chip comprising a laser source and scanning means for scanning, on the surface of the chip, a laser beam coming from the laser source. The device comprises means for constituting information (I) relating to the position of at least one stud of the chip with respect to the lighting means and means for constituting a control signal (B) of the scanning means from of said information (I).
L'invention concerne également une puce biologique active comprenant au moins une ligne de M plots (P), M étant un nombre entier supérieur ou égal à 1. Chaque plot est entouré d'une zone réfléchissante (Z).The invention also relates to an active biological chip comprising at least one line of M pads (P), M being an integer greater than or equal to 1. Each pad is surrounded by a reflecting zone (Z).
Un dispositif de lecture de puce biologique selon l'invention est un système miniaturisé facilement transportable, utilisable aussi bien hors laboratoire qu ' en laboratoire et permettant 1 ' obtention d'excellente performances.A biological chip reading device according to the invention is an easily transportable miniaturized system, usable both outside the laboratory and in the laboratory and allowing excellent performance to be obtained.
Les moyens pour illuminer la puce comprennent une source laser, des moyens optiques pour mettre en forme un faisceau lumineux issu de la source laser, des moyens pour diriger le faisceau laser vers la puce et des moyens de balayage du faisceau laser à la surface de la puce. Avantageusement, le circuit imprimé comprend des circuits électroniques pour alimenter et contrôler, entre autres, la source laser, les moyens photodétecteurs, les moyens de balayage, ainsi qu'une zone mémoire pour stocker les données issues des moyens de détection de la fluorescence.The means for illuminating the chip comprise a laser source, optical means for shaping a light beam coming from the laser source, means for directing the laser beam towards the chip and means for scanning the laser beam on the surface of the chip. Advantageously, the printed circuit comprises electronic circuits for supplying and controlling, inter alia, the laser source, the photodetector means, the scanning means, as well as a memory area for storing the data coming from the fluorescence detection means.
Selon l'invention, la puce biologique est une puce active. Par "puce active", il faut entendre une puce sur laquelle sont intégrés les photodétecteurs qui reçoivent l'émission de fluorescence. La puce active peut également contenir des mémoires informatiques pour enregistrer des informations. L'intégration des photodétecteurs à la puce biologique permet avantageusement de résoudre les problèmes d'alignement, de perte de lumière et de filtrage optique que l'on rencontre lorsque les photodétecteurs sont séparés de la puce.According to the invention, the biological chip is an active chip. By "active chip" is meant a chip on which are integrated the photodetectors which receive the fluorescence emission. The active chip can also contain computer memories for recording information. The integration of photodetectors with the biological chip advantageously makes it possible to solve the problems of alignment, loss of light and optical filtering which are encountered when the photodetectors are separated from the chip.
Selon l'invention, le champ balayé par le faisceau lumineux est plus grand que la superficie de la puce. Il n'est alors pas nécessaire de déplacer la puce par rapport aux moyens d'éclairage. Afin que s'effectue correctement le balayage de la puce, il est ainsi suffisant d'effectuer un repérage des plots.According to the invention, the field scanned by the light beam is larger than the area of the chip. It is then not necessary to move the chip relative to the lighting means. In order for the chip to be scanned correctly, it is therefore sufficient to mark the studs.
Avantageusement, la puce active est jetable. Le système de lecture comprend alors, d'une part, une tête de lecture qui n'est pas jetable constituée de moyens d'éclairage et de moyens de balayage et, d'autre part, des moyens de photodétection jetables intégrés à la puce active. Brève description des figuresAdvantageously, the active chip is disposable. The reading system then comprises, on the one hand, a reading head which is not disposable consisting of lighting means and scanning means and, on the other hand, disposable photodetection means integrated into the active chip . Brief description of the figures
D'autres caractéristiques et avantages de l'invention apparaîtront à la lecture d'un mode de réalisation préférentiel de l'invention fait en référence aux figures ci-annexées parmi lesquelles :Other characteristics and advantages of the invention will appear on reading a preferred embodiment of the invention made with reference to the appended figures among which:
- les figures lA et IB représentent un dispositif de lecture de puce biologique selon 1 ' invention,FIGS. 1A and 1B represent a device for reading a biological chip according to the invention,
- la figure 2 représente un plot de puce biologique selon un mode de réalisation particulier de- Figure 2 shows a biological chip pad according to a particular embodiment of
1 ' invention.1 invention.
Description détaillée de modes de mise en oeuyre de 1 ' invention Sur toutes les figures, les mêmes références désignent les mêmes éléments.Detailed description of methods of implementing the invention In all the figures, the same references designate the same elements.
Les figures 1A et IB représentent un dispositif de lecture de puce biologique selon l'invention.Figures 1A and IB show a biological chip reading device according to the invention.
La figure 1A représente une vue en perspective du dispositif.FIG. 1A represents a perspective view of the device.
Le dispositif de lecture comprend un circuit imprimé 1 et un bloc optique 2. Le circuit imprimé 1 est muni d'un connecteur 5 dans lequel vient s'enficher, lors de la lecture de la puce, un support 3 sur lequel la puce 4 est positionnée. Le bloc optique 2 comprend des moyens pour illuminer la puce et des moyens pour détecter la lumière réfléchie et/ou diffusée issue de la puce. Le bloc optique 2 est décrit plus en détail en figure IB. Selon l'invention, le bloc optique 2 vient s'enficher dans le circuit imprimé 1 et le support 3 dans le connecteur 5. Avantageusement, il n'est pas nécessaire que la précision du positionnement du bloc optique dans le circuit imprimé de même que la précision du positionnement du support 3 dans le connecteur 5 soient supérieures à l/10eme de millimètre. Le dispositif de lecture selon l'invention présente avantageusement un encombrement très réduit ainsi qu'une grande maniabilité. L'encombrement d'un tel système peut être, par exemple, de l'ordre de 200x100x100 mm3.The reading device comprises a printed circuit 1 and an optical unit 2. The printed circuit 1 is provided with a connector 5 into which is plugged, during the reading of the chip, a support 3 on which the chip 4 is positioned. The optical unit 2 comprises means for illuminating the chip and means for detecting the reflected and / or scattered light coming from the chip. The optical unit 2 is described in more detail in FIG. 1B. According to the invention, the optical unit 2 is plugged into the printed circuit 1 and the support 3 in the connector 5. Advantageously, it is not necessary that the precision of the positioning of the optical unit in the printed circuit as well as the precision of the positioning of the support 3 in the connector 5 be greater than 1/10 th of a millimeter. The reading device according to the invention advantageously has a very small footprint as well as great maneuverability. The size of such a system can be, for example, of the order of 200 × 100 × 100 mm 3 .
La figure IB représente une vue détaillée du bloc optique 2 en liaison avec une puce biologique 4.FIG. 1B represents a detailed view of the optical unit 2 in connection with a biological chip 4.
Le bloc optique 2 comprend un moyen d'éclairage 6 comprenant une source laser, des moyens 7 de balayage, selon deux dimensions, d'un faisceau issu de la source laser, des moyens 8 pour mettre en forme le faisceau issu des moyens de balayage 7, des moyens 9, 10 pour diriger et focaliser le faisceau laser vers la puce 4, une optique de détection 11 et des moyens photodétecteurs 12 pour recevoir et détecter la lumière renvoyée par la puce 4.The optical unit 2 comprises a lighting means 6 comprising a laser source, means 7 for scanning, in two dimensions, of a beam coming from the laser source, means 8 for shaping the beam coming from the scanning means 7, means 9, 10 for directing and focusing the laser beam towards the chip 4, a detection optic 11 and photodetector means 12 for receiving and detecting the light returned by the chip 4.
Selon le mode de réalisation préférentiel de l'invention, les moyens de balayage 7 sont constitués d'un déflecteur acousto-optique. Le balayage de la puce permet que les plots soient illuminés les uns après les autres. Il n'est alors pas nécessaire que la source laser émette un signal de puissance élevée. L'illumination d'un plot de 50 μm de côté peut être effectuée dans d'excellentes conditions avec un signal de 0,25 mW de puissance. Le balayage de la puce permet, par ailleurs, d'améliorer le rapport signal sur bruit.According to the preferred embodiment of the invention, the scanning means 7 consist of an acousto-optical deflector. The scanning of the chip allows the studs to be illuminated one after the other. The laser source does not therefore need to emit a high power signal. The illumination of a 50 μm side pad can be carried out under excellent conditions with a signal of 0.25 mW of power. Scanning the chip also improves the signal-to-noise ratio.
A titre d'exemple non limitatif, le faisceau laser que contient le moyen d'éclairage 6 peut avoir une puissance sensiblement inférieure à 5 mW et un diamètre de l'ordre de 150 μm.By way of nonlimiting example, the laser beam contained in the lighting means 6 may have a power substantially less than 5 mW and a diameter of the order of 150 μm.
Selon l'invention, le signal lumineux renvoyé par la puce 4 contient une information relative à la position des plots par rapport aux moyens d'éclairage. Les moyens photodétecteurs 12 comprennent des moyens pour convertir 1 ' information lumineuse relative à la position des plots en une information électrique I transmise à un circuit de traitement 13. Le signal S issu du circuit de traitement 13 est transmis à un circuit 14 de commande des moyens de balayage 7 qui reçoit, par ailleurs un signal de consigne C. Le signalAccording to the invention, the light signal returned by the chip 4 contains information relating to the position of the pads relative to the lighting means. The photodetector means 12 comprise means for converting the light information relating to the position of the pads into electrical information I transmitted to a processing circuit 13. The signal S from the processing circuit 13 is transmitted to a circuit 14 for controlling the scanning means 7 which also receives a setpoint signal C. The signal
S et le signal de consigne C sont comparés l'un à l'autre. Le circuit 14 de commande des moyens de balayage 7 délivre un signal de commande de balayage B à partir du signal d'erreur issu de la comparaison du signal S et du signal de consigne C. Il est alors possible d'obtenir un alignement des plots avec les moyens d'éclairage d'une très grande précision. Une précision inférieure à 5 μm peut ainsi être obtenue pour des plots de 50 μm de côté.S and the setpoint signal C are compared with each other. The circuit 14 for controlling the scanning means 7 delivers a scanning control signal B from the error signal resulting from the comparison of the signal S and the reference signal C. It is then possible to obtain an alignment of the pads with very precise lighting means. An accuracy of less than 5 μm can thus be obtained for studs with a side of 50 μm.
Le circuit de traitement 13 et le circuit 14 de commande des moyens de balayage 7 sont préférentiellement réalisés sur le circuit imprimé 1.The processing circuit 13 and the circuit 14 for controlling the scanning means 7 are preferably produced on the printed circuit 1.
Selon un premier mode de réalisation de l'invention, les moyens d'éclairage 6 comprennent, en plus de la source laser, au moins une diode électroluminescente pour éclairer la puce 4 en tout ou partie.According to a first embodiment of the invention, the lighting means 6 comprise, in addition to the laser source, at least one diode light-emitting to illuminate the chip 4 in whole or in part.
L'optique de détection 11 comprend alors une matrice de microlentilles et les moyens de détection 12 comprennent une matrice CCD (l'acronyme CCD provenant de l'anglais "Charge Coupled Device").The detection optic 11 then comprises a matrix of microlenses and the detection means 12 comprise a CCD matrix (the acronym CCD from the English "Charge Coupled Device").
L ' image formée par la matrice CCD du fait de l'illumination de la puce par la lumière issue de la diode électroluminescente présente un effet de moiré au cas où la trame définie par les microlentilles est décalée par rapport à la trame définie par les plots de la puce. Le signal électrique qui traduit l'effet de moiré constitue, selon le premier mode de réalisation de l'invention, l'information I qui représente le désalignement entre la puce 4 et les moyens d'éclairage 6.The image formed by the CCD matrix due to the illumination of the chip by the light emitted by the light-emitting diode exhibits a moire effect in the case where the frame defined by the microlenses is offset with respect to the frame defined by the pads of the chip. The electrical signal which translates the moiré effect constitutes, according to the first embodiment of the invention, the information I which represents the misalignment between the chip 4 and the lighting means 6.
Le circuit 13 de traitement de l'information électrique I permet de stocker 1 ' image de la puce 4 et de repérer la position des plots de la puce dans un repère absolu. Le signal de consigne C est alors représentatif de la position que doivent occuper les plots pour assurer l'alignement idéal entre les moyens d'éclairage de la puce et la puce.The circuit 13 for processing the electrical information I makes it possible to store the image of the chip 4 and to locate the position of the pads of the chip in an absolute reference. The setpoint signal C is then representative of the position that the pads must occupy to ensure ideal alignment between the lighting means of the chip and the chip.
Selon un deuxième mode de réalisation de l'invention, c'est la lumière diffusée issue de la puce biologique qui est utilisée pour aligner les plots avec les moyens d'éclairage.According to a second embodiment of the invention, it is the scattered light coming from the biological chip which is used to align the pads with the lighting means.
Les moyens de détection 12 comprennent alors une photodiode pour détecter l'amplitude de la lumière diffusée par un plot de la puce. La position correcte d'un plot par rapport aux moyens d'éclairage est alors détectée par un maximum de la lumière diffusée à la longueur d'onde d'excitation. L'information I est alors constituée du signal électrique détecté d'amplitude maximum.The detection means 12 then comprise a photodiode for detecting the amplitude of the light scattered by a pad of the chip. The correct position a pad relative to the lighting means is then detected by a maximum of the light scattered at the excitation wavelength. Information I then consists of the detected electrical signal of maximum amplitude.
Cette technique d'alignement est particulièrement avantageuse lorsque les plots des puces sont relativement espacés comme c'est le cas, par exemple, pour les puces MICAM ®.This alignment technique is particularly advantageous when the studs of the chips are relatively spaced, as is the case, for example, for MICAM® chips.
Selon un troisième mode de réalisation de l'invention, chaque plot P de puce biologique est entouré d'une zone réfléchissante Z. La zone Z est préférentiellement de forme circulaire comme représenté en figure 2. La zone Z est métallisée de façon à réfléchir la lumière qu'elle reçoit.According to a third embodiment of the invention, each pad P of a biological chip is surrounded by a reflecting zone Z. The zone Z is preferably of circular shape as shown in FIG. 2. The zone Z is metallized so as to reflect the light it receives.
Les moyens de détection 12 comprennent alors un détecteur quatre-quadrants sur lequel se forme 1 ' image de la zone Z . Après la recherche d'un plot de référence et compte-tenu de l'erreur de positionnement de la puce par rapport à ce plot de référence, les moyens de balayage 7 sont programmés pour amener l'éclairage de la puce en regard d'un plot P de façon qu'une image du plot se forme sur le détecteur quatre-quadrants.The detection means 12 then comprise a four-quadrant detector on which the image of the zone Z is formed. After the search for a reference pad and taking into account the positioning error of the chip relative to this reference pad, the scanning means 7 are programmed to bring the lighting of the chip opposite a plot P so that an image of the plot is formed on the four-quadrant detector.
De façon connue en soi, le détecteur quatre- quadrants permet alors de créer deux signaux d'erreur de positionnement, un premier signal d'erreur selon une première direction et un deuxième signal d'erreur selon une deuxième direction perpendiculaire à la première direction. Les deux signaux d'erreur constituent l'information I transmise au circuit de traitement 13.In a manner known per se, the four-quadrant detector then makes it possible to create two positioning error signals, a first error signal in a first direction and a second error signal in a second direction perpendicular to the first direction. The two error signals constitute the information I transmitted to the processing circuit 13.
Ce troisième mode de réalisation de 1 ' invention présente l'avantage de maintenir l'asservissement des moyens d'éclairage sur un plot alors même qu'une perturbation mécanique peut intervenir au niveau du lecteur. Par ailleurs, il n'est pas nécessaire que le déplacement du faisceau lumineux d'un plot à l'autre s'effectue de façon précise. This third embodiment of the invention has the advantage of maintaining the enslavement of the lighting means on a pad even though a mechanical disturbance may occur at the level of the reader. Furthermore, it is not necessary that the movement of the light beam from one stud to another takes place in a precise manner.

Claims

REVENDICATIONS
1. Dispositif de lecture de puce biologique active (4) comprenant au moins une ligne de M plots, M étant un nombre entier supérieur ou égal à 1, le dispositif comprenant un bloc optique (2) constitué de moyens d'éclairage (6, 7, 8, 9) de la puce comprenant une source laser (6) et des moyens de balayage (7) pour balayer, à la surface de la puce, un faisceau laser issu de la source laser (6), caractérisé en ce qu'il comprend des moyens pour constituer une information (I) relative à la position d'au moins un plot de la puce par rapport aux moyens d'éclairage (6, 7, 8, 9) et des moyens (13, 14) pour constituer un signal de commande (B) des moyens de balayage (7) à partir de ladite information (I).1. Device for reading an active biological chip (4) comprising at least one line of M pads, M being an integer greater than or equal to 1, the device comprising an optical unit (2) consisting of lighting means (6, 7, 8, 9) of the chip comprising a laser source (6) and scanning means (7) for scanning, on the surface of the chip, a laser beam coming from the laser source (6), characterized in that '' it comprises means for constituting information (I) relating to the position of at least one stud of the chip with respect to the lighting means (6, 7, 8, 9) and means (13, 14) for constitute a control signal (B) of the scanning means (7) from said information (I).
2. Dispositif selon la revendication 1, caractérisé en ce que les moyens pour constituer une information (I) comprennent au moins une diode électroluminescente pour éclairer la puce en tout ou partie, une matrice de microlentilles pour recevoir la lumière issue de la diode électroluminescente renvoyée par la puce et une matrice CCD.2. Device according to claim 1, characterized in that the means for constituting information (I) comprise at least one light-emitting diode to illuminate the chip in whole or in part, a matrix of microlenses to receive the light coming from the light-emitting diode returned by the chip and a CCD matrix.
3. Dispositif selon la revendication 1, caractérisé en ce que les moyens pour constituer une information ( I ) comprennent une photodiode pour détecter l'amplitude de la lumière diffusée par un plot de la puce. 3. Device according to claim 1, characterized in that the means for constituting information (I) comprise a photodiode for detecting the amplitude of the light scattered by a pad of the chip.
4. Dispositif selon la revendication 1, caractérisé en ce que les moyens pour constituer une information (I) comprennent un détecteur quatre quadrants pour détecter la lumière réfléchie par une zone métallisée réfléchissante (Z) entourant un plot (P) de la puce.4. Device according to claim 1, characterized in that the means for constituting information (I) comprise a four quadrant detector for detecting the light reflected by a reflective metallized area (Z) surrounding a pad (P) of the chip.
5. Dispositif selon l'une quelconque des revendications précédentes, caractérisé en ce que les moyens (13, 14) pour constituer un signal de commande (B) des moyens de balayage (7) comprennent un circuit de traitement (13) pour délivrer un signal traité (S) à partir de ladite information (I) et un circuit de commande (14) recevant, d'une part, ledit signal traité et, d'autre part, un signal de consigne (C).5. Device according to any one of the preceding claims, characterized in that the means (13, 14) for constituting a control signal (B) of the scanning means (7) comprise a processing circuit (13) for delivering a signal processed (S) from said information (I) and a control circuit (14) receiving, on the one hand, said processed signal and, on the other hand, a setpoint signal (C).
6. Dispositif selon l'une quelconque des revendications précédentes, caractérisé en ce qu'il comprend un circuit imprimé (1) muni d'un connecteur (5) dans lequel vient s'enficher, lors de la lecture de la puce (4), un support (3) sur lequel la puce (4) est positionnée, le bloc optique (2) étant enfichable dans le circuit imprimé de façon à placer les moyens d'éclairage (6, 7, 8, 9) en position pour éclairer la puce (4 ) .6. Device according to any one of the preceding claims, characterized in that it comprises a printed circuit (1) provided with a connector (5) in which is plugged in, when reading the chip (4) , a support (3) on which the chip (4) is positioned, the optical unit (2) being pluggable into the printed circuit so as to place the lighting means (6, 7, 8, 9) in position for lighting the chip (4).
7. Dispositif selon la revendication 6, caractérisé en ce que le circuit de traitement (13) et le circuit de commande (14) sont réalisés sur le circuit imprimé (1). 7. Device according to claim 6, characterized in that the processing circuit (13) and the control circuit (14) are produced on the printed circuit (1).
8. Dispositif selon l'une quelconque des revendications précédentes, caractérisé en ce que les moyens de balayage (7) sont constitués d'un déflecteur acousto-optique .8. Device according to any one of the preceding claims, characterized in that the scanning means (7) consist of an acousto-optical deflector.
9. Puce biologique active comprenant au moins une ligne de M plots (P), M étant un nombre entier supérieur ou égal à 1, caractérisée en ce que chaque plot (P) est entouré d'une zone (Z) réfléchissante. 9. Active biological chip comprising at least one line of M pads (P), M being an integer greater than or equal to 1, characterized in that each pad (P) is surrounded by a reflective zone (Z).
PCT/FR2000/002143 1999-07-27 2000-07-26 Biochip reading device WO2001007895A1 (en)

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