[go: up one dir, main page]

WO2004011938B1 - Method and device for screening molecules in cells - Google Patents

Method and device for screening molecules in cells

Info

Publication number
WO2004011938B1
WO2004011938B1 PCT/FR2003/002298 FR0302298W WO2004011938B1 WO 2004011938 B1 WO2004011938 B1 WO 2004011938B1 FR 0302298 W FR0302298 W FR 0302298W WO 2004011938 B1 WO2004011938 B1 WO 2004011938B1
Authority
WO
WIPO (PCT)
Prior art keywords
support
drops
cell
aqueous
reagent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/FR2003/002298
Other languages
French (fr)
Other versions
WO2004011938A3 (en
WO2004011938A2 (en
Inventor
Beatrice Schaack
Francois Chatelain
Brigitte Fouque
Alexandra Fuchs
Yves Fouillet
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Commissariat a lEnergie Atomique et aux Energies Alternatives CEA
Original Assignee
Commissariat a lEnergie Atomique CEA
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 Commissariat a lEnergie Atomique CEA filed Critical Commissariat a lEnergie Atomique CEA
Priority to EP03750828A priority Critical patent/EP1525472A2/en
Priority to CA002492933A priority patent/CA2492933A1/en
Priority to JP2004523860A priority patent/JP2005533509A/en
Priority to AU2003269041A priority patent/AU2003269041A1/en
Publication of WO2004011938A2 publication Critical patent/WO2004011938A2/en
Publication of WO2004011938A3 publication Critical patent/WO2004011938A3/en
Publication of WO2004011938B1 publication Critical patent/WO2004011938B1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5023—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects on expression patterns
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5032—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects on intercellular interactions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5058—Neurological cells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00364—Pipettes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00364—Pipettes
    • B01J2219/00367—Pipettes capillary
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00378—Piezoelectric or ink jet dispensers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00646—Making arrays on substantially continuous surfaces the compounds being bound to beads immobilised on the solid supports
    • B01J2219/0065—Making arrays on substantially continuous surfaces the compounds being bound to beads immobilised on the solid supports by the use of liquid beads
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
    • 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
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1034—Transferring microquantities of liquid
    • G01N2035/1046—Levitated, suspended drops

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Physics & Mathematics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Microbiology (AREA)
  • Toxicology (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention concerns a method for reacting a reagent R with a cell C, which consists in: setting the cell C on a support S comprising a substantially planar surface, in the form of an aqueous drop on said surface; covering the planar surface of the support S whereon the aqueous drop has been set containing the cell C with a separation film F, allowing through gases and preventing the aqueous drops set on the support S from evaporating, F being non-miscible with the reagent R; triggering the reaction between the reagent R and the cell C by introducing the reagent R in the aqueous drop containing the cell C. The invention also concerns a device for implementing said procedure and its applications.

Claims

40 40 REVENDICATIONS MODIFIEESMODIFIED CLAIMS [reçues par le bureau international le 06 août 2004 (06.08.04); revendications originales 1-59 remplacées par de nouvelles revendications 1-56 (7 pages)][received by the International Bureau on 06 August 2004 (06.08.04); original claims 1-59 replaced by new claims 1-56 (7 pages)] REVENDICATIONS 1°) Procédé de mise en réaction d'un réactif R avec au moins une cellule C, ledit procédé étant caractérisé en ce que : - la cellule C est déposée sur un support S comportant une surface sensiblement plane, sous forme d'une goutte aqueuse sur ladite surface,1) Method for reacting a reagent R with at least one cell C, said method being characterized in that: the cell C is deposited on a support S comprising a substantially flat surface in the form of a drop aqueous on said surface, - la surface sensiblement plane du support S sur laquelle a été déposée la goutte aqueuse contenant la cellule C est recouverte par un film de séparation F, permettant le passage des gaz et empêchant l'évaporation des gouttes aqueuses déposées sur le support S, F étant non miscible avec le réactif R,- The substantially flat surface of the support S on which was deposited the aqueous drop containing the cell C is covered by a separating film F, allowing the passage of gases and preventing the evaporation of aqueous drops deposited on the support S, F being immiscible with reagent R, - la réaction entre le réactif R et la cellule C est déclenchée par l'introduction du réactif R dans la goutte aqueuse contenant la cellule C,the reaction between the reagent R and the cell C is triggered by the introduction of the reagent R into the aqueous droplet containing the cell C, - le support S présente sur sa surface plane un caractère hydrophobe et au moins un moyen destiné à la réception des gouttes aqueuses constitué par une ou plusieurs zones hydrophiles.- The support S has on its flat surface a hydrophobic character and at least one means for receiving aqueous drops consisting of one or more hydrophilic zones. 2°) Procédé selon la revendication 1, caractérisé en ce que l'accrochage des gouttes sur le support S se fait par capillarité.2) Method according to claim 1, characterized in that the attachment of the drops on the support S is by capillarity. 3°) Procédé selon la revendication 1 ou de la revendication 2, caractérisé en ce que le support S est constitué par une plaque d'un matériau choisi parmi le silicium, le verre ou un polymère.3) Method according to claim 1 or claim 2, characterized in that the support S is constituted by a plate of a material selected from silicon, glass or a polymer. 4°) Procédé selon l'une quelconque des revendications 1 à 3, caractérisé en ce qu'une goutte aqueuse contenant la cellule C est déposée sur le support S, une seconde goutte aqueuse contenant le réactif R est injecté, à l'aide de tout ou moyen d'injection approprié, directement dans la goutte contenant la cellule C. 5°) Procédé selon l'une quelconque des revendications 1 à 3, caractérisé en ce qu'une première goutte aqueuse est déposée sur le support S puis une seconde goutte aqueuse est déposée sur le même support à proximité de la première, l'une de ces gouttes contient la cellule C, l'autre le réactif R, la réaction du réactif R avec la cellule C est déclenchée par la fusion des deux gouttes. 6°) Procédé selon l'une quelconque des revendications 1 à 3, caractérisé en ce que le réactif R est fixé au support S ou au film F, la cellule C est 414) Method according to any one of claims 1 to 3, characterized in that an aqueous drop containing the cell C is deposited on the support S, a second aqueous drop containing the reagent R is injected with the aid of all or appropriate injection means, directly in the drop containing the cell C. 5 °) Method according to any one of claims 1 to 3, characterized in that a first aqueous drop is deposited on the support S and then a second The aqueous drop is deposited on the same support near the first, one of these drops contains the cell C, the other the reagent R, the reaction of the reagent R with the cell C is triggered by the melting of the two drops. Process according to any one of Claims 1 to 3, characterized in that the reagent R is attached to the support S or to the film F, the cell C is 41 déposée sous forme d'une goutte aqueuse sur le support S et le réactif R est alors décroché du support S ou du film F afin de permettre sa réaction avec la cellule.deposited in the form of an aqueous drop on the support S and the reagent R is then unhooked from the support S or the film F to allow its reaction with the cell. 7°) Procédé selon l'une quelconque des revendications 1 à 6, caractérisé en ce que le film de séparation F est un liquide choisi parmi les huiles et les solvants organiques.7 °) Method according to any one of claims 1 to 6, characterized in that the separating film F is a liquid selected from oils and organic solvents. 8°) Procédé selon la revendication 7, caractérisé en ce que le film de séparation F est choisi parmi les huiles minérales et les huiles de silicone.8 °) Method according to claim 7, characterized in that the separating film F is selected from mineral oils and silicone oils. 9°) Procédé selon l'une quelconque des revendications 1 à 6, caractérisé en ce que le film de séparation F est de l'air saturé en humidité. 10°) Procédé selon Tune des revendications 1 à 6, caractérisé en ce que le film de séparation F est un film souple, solide.9 °) Method according to any one of claims 1 to 6, characterized in that the separating film F is air saturated with moisture. 10 °) Method according to one of claims 1 to 6, characterized in that the separating film F is a flexible film, solid. 11°) Procédé selon la revendication 10, caractérisé en ce que le film de séparation F est en polydiméthylsiloxane ou en nitrocellulose.11 °) A method according to claim 10, characterized in that the separating film F is polydimethylsiloxane or nitrocellulose. 12°) Procédé selon l'une quelconque des revendications 1 à 6, caractérisé en ce que le film de séparation F est un capot rigide alvéolé en matériau poreux.12 °) Method according to any one of claims 1 to 6, characterized in that the separating film F is a rigid honeycomb bonnet of porous material. 13°) Procédé selon l'une quelconque des revendications 1 à 12 caractérisé en ce que le dépôt des gouttes aqueuses contenant une ou plusieurs cellules ou un réactif sur le support S se fait au moyen de fins capillaires. 14°) Procédé selon l'une quelconque des revendications 1 à 12 caractérisé en ce que le dépôt des gouttes aqueuses contenant une ou plusieurs cellules ou un réactif sur le support S se fait au moyen d'une buse.13 °) Method according to any one of claims 1 to 12 characterized in that the deposition of aqueous drops containing one or more cells or a reagent on the support S is by means of fine capillaries. 14 °) Method according to any one of claims 1 to 12 characterized in that the deposition of aqueous drops containing one or more cells or a reagent on the support S is by means of a nozzle. 15°) Procédé selon l'une quelconque des revendications 1 à 14, caractérisé en ce qu'il comporte une étape de déplacement du support S après le dépôt sur le support S de la première série de gouttes.15 °) Method according to any one of claims 1 to 14, characterized in that it comprises a step of moving the support S after the deposition on the support S of the first series of drops. 16°) Procédé selon l'une quelconque des revendications 1 à 15, caractérisé en ce que les cultures de cellules sous forme de gouttes aqueuses sont conservées au moins 24 heures.16 °) Method according to any one of claims 1 to 15, characterized in that the cultures of cells in the form of aqueous drops are stored for at least 24 hours. 17°) Procédé selon l'une quelconque des revendications 1 à 16, caractérisé en ce que plusieurs gouttes aqueuses comprenant chacune au moins une cellule sont déposées sur le support S, sous le film de séparation F, lesdites gouttes étant isolées les unes des autres. 4217 °) Method according to any one of claims 1 to 16, characterized in that several aqueous drops each comprising at least one cell are deposited on the support S, under the separating film F, said drops being isolated from each other . 42 18°) Procédé selon l'une quelconque des revendications 1 à 17, caractérisé en ce qu'une goutte contient de 1 à 100 cellules18 °) Method according to any one of claims 1 to 17, characterized in that a drop contains from 1 to 100 cells 19°) Procédé selon l'une quelconque des revendications 17 et 18, caractérisé en ce que l'on place des cellules différentes dans les différentes gouttes. 20°) Procédé selon l'une quelconque des revendications 17 et 18, caractérisé en ce que l'on place des cellules identiques dans les différentes gouttes.19 °) Method according to any one of claims 17 and 18, characterized in that one places different cells in the different drops. 20 °) Method according to any one of claims 17 and 18, characterized in that one places identical cells in the different drops. 21°) Procédé selon la revendication 20, caractérisé en ce que le support est une plaque hydrophobe comportant des zones hydrophiles et que l'étape d'injection des gouttes aqueuses contenant des cellules est remplacé par l'immersion de la plaque dans une solution aqueuse contenant les cellules.21 °) Method according to claim 20, characterized in that the support is a hydrophobic plate having hydrophilic zones and that the step of injecting the aqueous drops containing cells is replaced by immersing the plate in an aqueous solution containing the cells. 22°) Procédé selon l'une quelconque des revendications 1 à 21, caractérisé en ce que les molécules de réactif sont préparées directement après dépôt sur le support, par un procédé choisi parmi la synthèse in situ, la transcription in vitro dans la goutte, la réaction de polymérisation en chaîne peptidique et nucléique. 23°) Procédé selon l'une quelconque des revendications 1 à 22, caractérisé en ce que le réactif est une molécule d'ADN.22 °) A method according to any one of claims 1 to 21, characterized in that the reagent molecules are prepared directly after deposition on the support, by a method selected from in situ synthesis, in vitro transcription in the drop, the peptide and nucleic chain polymerization reaction. 23 °) Method according to any one of claims 1 to 22, characterized in that the reagent is a DNA molecule. 24°) Procédé selon la revendications 23, caractérisé en ce que l'ADN est sous forme précipitée, notamment sous forme de phosphate de calcium.24 °) Method according to claim 23, characterized in that the DNA is in precipitated form, especially in the form of calcium phosphate. 25°) Procédé selon l'une quelconque des revendications 1 à 22, caractérisé en ce que le réactif est facteur de transcription.25 °) Method according to any one of claims 1 to 22, characterized in that the reagent is transcription factor. 26°) Procédé selon l'une quelconque des revendications 1 à 25, caractérisé en ce que l'on dépose successivement plusieurs réactifs destinés à réagir avec une même cellule.26 °) Method according to any one of claims 1 to 25, characterized in that one deposits successively several reagents for reacting with the same cell. 27°) Procédé selon l'une quelconque des revendications 1 à 26, caractérisé en ce que l'on dépose plusieurs gouttes aqueuses renfermant des cellules et on fait fusionner ces gouttes.27 °) A method according to any one of claims 1 to 26, characterized in that is deposited several aqueous drops containing cells and is fused these drops. 28°) Procédé selon la revendication 27, caractérisé en ce que l'on dépose des cellules gliales et des neurones pour les faire communiquer au sein d'une même goutte. 29°) Procédé selon l'une quelconque des revendications 1 à 28, caractérisé en ce que l'on fait réagir des réactifs dans un premier type de cellules de façon à déclencher une réaction cellulaire, telle que la production d'une protéine recombinante puis on fait réagir cette première cellule avec une cellule d'un autre type par fusion avec une autre goutte.28 °) Method according to claim 27, characterized in that glial cells and neurons are deposited to make them communicate within a single drop. 29 °) A method according to any one of claims 1 to 28, characterized in that reagents are reacted in a first type of cells so as to trigger a cellular reaction, such as the production of a protein recombinant then this first cell is reacted with a cell of another type by melting with another drop. 30°) Procédé selon l'une quelconque des revendications 1 à 29 dans lequel le support comporte des moyens de séparation, caractérisé en ce que l'on dépose une goutte aqueuse comprenant au moins une cellule d'un premier type d'un coté du moyen de séparation et une goutte aqueuse comprenant au moins une cellule d'un second type de l'autre côté du moyen de séparation puis que l'on opère la fusion des gouttes cellulaires.30 °) Method according to any one of claims 1 to 29 wherein the support comprises separation means, characterized in that is deposited an aqueous drop comprising at least one cell of a first type on one side of the separating means and an aqueous drop comprising at least one cell of a second type on the other side of the separating means and then the melting of the cell drops is performed. 31°) Procédé selon l'une quelconque des revendications 1 à 30, caractérisé en ce que le réactif est choisi parmi les molécules marquées, notamment les marqueurs fluorescents et radioactifs.31 °) Method according to any one of claims 1 to 30, characterized in that the reagent is selected from the labeled molecules, including fluorescent and radioactive markers. 32°) Procédé selon l'une quelconque des revendications 1 à 31, caractérisé en ce que la cellule est choisie parmi : des cellules primaires, des hybridomes, des lignées de cellules, des cellules souches, un morceau de tissu cellulaire, et leurs mélanges.32 °) Method according to any one of claims 1 to 31, characterized in that the cell is selected from: primary cells, hybridomas, cell lines, stem cells, a piece of cellular tissue, and mixtures thereof . 33°) Dispositif permettant la mise en réaction d'un réactif R avec une cellule C, ce dispositif étant caractérisé en ce qu'il comporte :33 °) Device for reacting a reagent R with a cell C, this device being characterized in that it comprises: - un support S comportant une surface sensiblement plane à caractère hydrophobe et au moins un moyen destiné à la réception des gouttes aqueuses, constitué d'une ou plusieurs zones hydrophiles, la surface S étant recouverte d'un film de séparation F permettant le passage des gaz et empêchant l'évaporation des gouttes aqueuses déposées sur le support, F étant non miscible avec le réactif R,a support S comprising a substantially flat surface of hydrophobic character and at least one means for receiving aqueous drops, consisting of one or more hydrophilic zones, the surface S being covered with a separation film F allowing the passage of gas and preventing the evaporation of the aqueous drops deposited on the support, F being immiscible with the reagent R, - des moyens permettant le dépôt sur ladite surface et sous le film F, de gouttes aqueuses contenant la cellule C, - une enceinte à atmosphère contrôlée dans laquelle est placé le support S de façon à permettre la survie de la cellule C.- Means for depositing on said surface and under the film F, aqueous drops containing the cell C - a controlled atmosphere chamber in which the support S is placed so as to allow the survival of the cell C. 34°) Dispositif selon la revendication 33, caractérisé en ce que le support S est constitué par une plaque d'un matériau choisi parmi le silicium, le verre ou un polymère. 35°) Dispositif selon la revendication 33 ou la revendication 34, caractérisé en ce que le moyen destiné à la réception des gouttes aqueuses consiste en des zones de la surface plane du support S d'une taille allant de 5 μm2 à 5 mm2. 4434 °) Device according to claim 33, characterized in that the support S is constituted by a plate of a material selected from silicon, glass or a polymer. 35 °) Device according to claim 33 or claim 34, characterized in that the means for receiving aqueous drops consists of areas of the flat surface of the support S with a size ranging from 5 microns 2 to 5 mm 2 . 44 36°) Dispositif selon l'une quelconque des revendications 33 à 35, caractérisé en ce que le support S présente au moins l'une des caractéristiques a) à c) ci-dessous : a) le support S comporte sur sa surface plane des cavités, d'une profondeur allant de 1 micron à 1 millimètre constituant le moyen de réception ; b) le support S est une plaque munie d'excroissances d'épaisseur allant de 1 micron à 1 millimètre, disposées sur sa surface et destinées à favoriser l'accrochage des gouttes ; c) le support S est une plaque munie d'au moins un fil, sur lequel viennent s'accrocher les gouttes.36 °) Device according to any one of claims 33 to 35, characterized in that the support S has at least one of the characteristics a) to c) below: a) the support S has on its flat surface cavities, of a depth ranging from 1 micron to 1 millimeter constituting the receiving means; b) the support S is a plate provided with excrescences of thickness ranging from 1 micron to 1 millimeter, arranged on its surface and intended to promote the attachment of the drops; c) the support S is a plate provided with at least one wire, on which hang the drops. 37°) Dispositif selon Tune quelconque des revendications 33 à 36, caractérisé en ce que les moyens permettant le dépôt des gouttes aqueuses sur le support S consistent en de fins capillaires.37 °) Device according to any one of claims 33 to 36, characterized in that the means for depositing aqueous drops on the support S consist of fine capillaries. 38°) Dispositif selon l'une quelconque des revendications 33 à 37, caractérisé en ce que les moyens permettant le dépôt des gouttes aqueuses sur le support S consistent en un système piézoélectrique muni d'une buse.38 °) Device according to any one of claims 33 to 37, characterized in that the means for depositing the aqueous drops on the support S consist of a piezoelectric system provided with a nozzle. 39°) Dispositif selon l'une quelconque des revendications 33 à 38, caractérisé en ce que le support S du dispositif est mobile.39 °) Device according to any one of claims 33 to 38, characterized in that the support S of the device is movable. 40°) Dispositif selon l'une quelconque des revendications 33 à 39, caractérisé en ce que le support S est constitué d'un film solide fixé à des rouleaux à ses deux extrémités, les rouleaux étant munis de moyens de bobinage de façon à permettre le déplacement du film et donc le déplacement des gouttes qui ont été déposées dessus.40 °) Device according to any one of claims 33 to 39, characterized in that the support S consists of a solid film attached to rollers at both ends, the rollers being provided with winding means so as to allow the displacement of the film and thus the displacement of the drops which have been deposited on it. 41°) Dispositif selon l'une quelconque des revendications 33 à 40, caractérisé en ce qu'il comporte en outre au moins un moyen choisi parmi :41 °) Device according to any one of claims 33 to 40, characterized in that it further comprises at least one means selected from: - des moyens permettant d'apporter de l'énergie à une ou plusieurs gouttes déposées sur le support ;means for supplying energy to one or more drops deposited on the support; - des moyens de traitement optique d'une ou plusieurs gouttes déposées sur le support ; - des moyens d'application d'un champ magnétique ou d'un champ électrique à une ou plusieurs gouttes déposées sur le support ; 45means for optically treating one or more drops deposited on the support; means for applying a magnetic field or an electric field to one or more drops deposited on the support; 45 - des moyens de détection focalisés sur une ou plusieurs gouttes déposées sur le support ;detection means focused on one or more drops deposited on the support; - des moyens destinés à favoriser la transfection.means for promoting transfection. 42°) Dispositif selon l'une quelconque des revendications 33 à 41, caractérisé en ce que les moyens utilisés dans le dispositif sont reliés à un dispositif de contrôle permettant son automatisation.42 °) Device according to any one of claims 33 to 41, characterized in that the means used in the device are connected to a control device for its automation. 43°) Dispositif selon l'une quelconque des revendications 33 à 42, caractérisé en ce que le support comprend des moyens de réception disposés de façon régulière sous forme de matrices. 44°) Dispositif selon l'une quelconque des revendications 33 à 43, caractérisé en ce que le support est muni de moyens de séparation permettant de séparer deux types de cellules distincts mais autorisant le passage de petites molécules entre ces cellules.43 °) Device according to any one of claims 33 to 42, characterized in that the support comprises receiving means regularly arranged in the form of dies. 44 °) Device according to any one of claims 33 to 43, characterized in that the support is provided with separation means for separating two distinct types of cells but allowing the passage of small molecules between these cells. 45°) Dispositif selon la revendication 44, caractérisé en ce que les moyens de séparation sont disposés au niveau des moyens de réception, sur le support.45 °) Device according to claim 44, characterized in that the separating means are arranged at the receiving means, on the support. 46°) Dispositif selon l'une quelconque des revendications 33 à 45, caractérisé en ce que les gouttes aqueuses contenant une ou plusieurs cellules comportent un milieu de culture.46 °) Device according to any one of claims 33 to 45, characterized in that the aqueous drops containing one or more cells comprise a culture medium. 47°) Dispositif selon l'une quelconque des revendications 33 à 46, caractérisé en ce que le support est doté d'une surface dont les propriétés d'hydrophilie/hydrophobie peuvent varier sous l'influence d'un paramètre tel que la température, un champ électrique, un champ magnétique, une irradiation.47 °) Device according to any one of claims 33 to 46, characterized in that the support is provided with a surface whose hydrophilic / hydrophobic properties may vary under the influence of a parameter such as temperature, an electric field, a magnetic field, an irradiation. 48°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour réaliser simultanément et de façon automatisée un grand nombre de réactions d'un réactif sur une cellule en faisant varier la nature du réactif et de la cellule.48 °) Use of a device according to any one of claims 33 to 47 to realize simultaneously and automatically a large number of reactions of a reagent on a cell by varying the nature of the reagent and the cell. 49°) Utilisation selon la revendication 48, pour réaliser le criblage d'un ensemble de composés chimiques sur des cellules vivantes.49 °) Use according to claim 48 for performing the screening of a set of chemical compounds on living cells. 50°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour étudier des systèmes cellulaires choisi paraii : les réseaux de neurones, Fépiderme. 4650 °) Use of a device according to any one of claims 33 to 47 to study cellular systems selected paraii: neural networks, the epidermis. 46 51°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour étudier l'action sur une cellule d'un réactif choisis parmi : les molécules d'acides nucléiques, les protéines, les peptides, les molécules d'acide nucléique peptidique. 52°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour l'expression de protéines recombinantes.51 °) Use of a device according to any one of claims 33 to 47 to study the action on a cell of a reagent chosen from: nucleic acid molecules, proteins, peptides, molecules of peptide nucleic acid. 52 °) Use of a device according to any one of claims 33 to 47 for the expression of recombinant proteins. 53°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour réaliser le criblage de molécules nucléiques destinées à modifier l'expression de gènes dans les cellules. 54°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour rechercher des séquences génomiques promotrices.53 °) Use of a device according to any one of claims 33 to 47 for performing the screening of nucleic molecules intended to modify the expression of genes in the cells. 54 °) Use of a device according to any one of claims 33 to 47 to search promoter genomic sequences. 55°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour étudier les interactions entre des cellules de différents types. 56°) Utilisation d'un dispositif selon l'une quelconque des revendications 33 à 47 pour préparation et le criblage d'ARNsi. 55 °) Use of a device according to any one of claims 33 to 47 to study the interactions between cells of different types. 56 °) Use of a device according to any one of claims 33 to 47 for the preparation and screening of siRNA.
PCT/FR2003/002298 2002-07-23 2003-07-21 Method and device for screening molecules in cells Ceased WO2004011938A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP03750828A EP1525472A2 (en) 2002-07-23 2003-07-21 Method and device for screening molecules in cells
CA002492933A CA2492933A1 (en) 2002-07-23 2003-07-21 Method and device for screening molecules in cells
JP2004523860A JP2005533509A (en) 2002-07-23 2003-07-21 Method and apparatus for screening molecules in cells
AU2003269041A AU2003269041A1 (en) 2002-07-23 2003-07-21 Method and device for screening molecules in cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR0209326A FR2842747B1 (en) 2002-07-23 2002-07-23 METHOD AND DEVICE FOR SCREENING MOLECULES IN CELLS
FR0209326 2002-07-23

Publications (3)

Publication Number Publication Date
WO2004011938A2 WO2004011938A2 (en) 2004-02-05
WO2004011938A3 WO2004011938A3 (en) 2004-08-05
WO2004011938B1 true WO2004011938B1 (en) 2004-09-16

Family

ID=30011408

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FR2003/002298 Ceased WO2004011938A2 (en) 2002-07-23 2003-07-21 Method and device for screening molecules in cells

Country Status (6)

Country Link
EP (1) EP1525472A2 (en)
JP (1) JP2005533509A (en)
AU (1) AU2003269041A1 (en)
CA (1) CA2492933A1 (en)
FR (1) FR2842747B1 (en)
WO (1) WO2004011938A2 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9476856B2 (en) 2006-04-13 2016-10-25 Advanced Liquid Logic, Inc. Droplet-based affinity assays
US10072241B2 (en) 2013-03-13 2018-09-11 Innovative Surface Technologies, Inc. Conical devices for three-dimensional aggregate(s) of eukaryotic cells

Families Citing this family (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2857451B1 (en) * 2003-07-11 2005-09-30 Commissariat Energie Atomique METHOD AND DEVICE FOR ANALYSIS OF LIVE REACTION ENVIRONMENTS
US20090215192A1 (en) * 2004-05-27 2009-08-27 Stratos Biosystems, Llc Solid-phase affinity-based method for preparing and manipulating an analyte-containing solution
EP1859330B1 (en) 2005-01-28 2012-07-04 Duke University Apparatuses and methods for manipulating droplets on a printed circuit board
DE112006000657T5 (en) * 2005-03-30 2008-02-07 National University Corporation Nagoya University, Nagoya Process for producing biological organic material and culture vessel therefor
WO2007063898A1 (en) * 2005-11-29 2007-06-07 The University Of Tokyo Method and apparatus for crystallization of protein, and apparatus for protein crystallization treatment
US20140193807A1 (en) 2006-04-18 2014-07-10 Advanced Liquid Logic, Inc. Bead manipulation techniques
US8809068B2 (en) 2006-04-18 2014-08-19 Advanced Liquid Logic, Inc. Manipulation of beads in droplets and methods for manipulating droplets
US10078078B2 (en) 2006-04-18 2018-09-18 Advanced Liquid Logic, Inc. Bead incubation and washing on a droplet actuator
US7439014B2 (en) 2006-04-18 2008-10-21 Advanced Liquid Logic, Inc. Droplet-based surface modification and washing
WO2009111769A2 (en) 2008-03-07 2009-09-11 Advanced Liquid Logic, Inc. Reagent and sample preparation and loading on a fluidic device
ES2423930T3 (en) 2007-02-09 2013-09-25 Advanced Liquid Logic, Inc. Droplet actuator devices and methods using magnetic beads
WO2009032863A2 (en) 2007-09-04 2009-03-12 Advanced Liquid Logic, Inc. Droplet actuator with improved top substrate
EP2237955A4 (en) 2007-12-23 2016-04-20 Advanced Liquid Logic Inc Droplet actuator configurations and methods of conducting droplet operations
WO2009137415A2 (en) 2008-05-03 2009-11-12 Advanced Liquid Logic, Inc. Reagent and sample preparation, loading, and storage
US8926065B2 (en) 2009-08-14 2015-01-06 Advanced Liquid Logic, Inc. Droplet actuator devices and methods
US9091649B2 (en) 2009-11-06 2015-07-28 Advanced Liquid Logic, Inc. Integrated droplet actuator for gel; electrophoresis and molecular analysis
EP2516669B1 (en) 2009-12-21 2016-10-12 Advanced Liquid Logic, Inc. Enzyme assays on a droplet actuator
WO2011142117A1 (en) 2010-05-11 2011-11-17 パナソニック株式会社 Cell culture substrate and cell culture method using same
US9188615B2 (en) 2011-05-09 2015-11-17 Advanced Liquid Logic, Inc. Microfluidic feedback using impedance detection
AU2012279420A1 (en) 2011-07-06 2014-01-30 Advanced Liquid Logic Inc Reagent storage on a droplet actuator
US9513253B2 (en) 2011-07-11 2016-12-06 Advanced Liquid Logic, Inc. Droplet actuators and techniques for droplet-based enzymatic assays
WO2013016413A2 (en) 2011-07-25 2013-01-31 Advanced Liquid Logic Inc Droplet actuator apparatus and system
WO2013078216A1 (en) 2011-11-21 2013-05-30 Advanced Liquid Logic Inc Glucose-6-phosphate dehydrogenase assays
CA2877950C (en) 2012-06-27 2021-06-22 Advanced Liquid Logic Inc. Techniques and droplet actuator designs for reducing bubble formation
EP3362177A1 (en) 2015-10-16 2018-08-22 Oxford University Innovation Limited Microfluidic arrangements
GB201518392D0 (en) * 2015-10-16 2015-12-02 Isis Innovation Microfluidic arrangements
GB201614150D0 (en) 2016-08-18 2016-10-05 Univ Oxford Innovation Ltd Microfluidic arrangements
CN114867556B (en) * 2019-11-06 2023-09-15 高丽大学校算学协力团 Droplet deformation-based method for delivering materials into cells and chips for it
JP7135049B2 (en) * 2020-10-12 2022-09-12 エコール ポリテクニック Methods for handling microdroplets containing samples

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AUPN923596A0 (en) * 1996-04-12 1996-05-09 Australian Biomedical Corporation Limited Method and apparatus for treatment of human or animal cell samples
AU6571598A (en) * 1997-03-18 1998-10-12 Chromaxome Corporation Methods for screening compounds using encapsulated cells
GB9808836D0 (en) * 1998-04-27 1998-06-24 Amersham Pharm Biotech Uk Ltd Microfabricated apparatus for cell based assays
FR2857451B1 (en) * 2003-07-11 2005-09-30 Commissariat Energie Atomique METHOD AND DEVICE FOR ANALYSIS OF LIVE REACTION ENVIRONMENTS

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9476856B2 (en) 2006-04-13 2016-10-25 Advanced Liquid Logic, Inc. Droplet-based affinity assays
US10072241B2 (en) 2013-03-13 2018-09-11 Innovative Surface Technologies, Inc. Conical devices for three-dimensional aggregate(s) of eukaryotic cells

Also Published As

Publication number Publication date
CA2492933A1 (en) 2004-02-05
WO2004011938A3 (en) 2004-08-05
AU2003269041A8 (en) 2004-02-16
JP2005533509A (en) 2005-11-10
EP1525472A2 (en) 2005-04-27
WO2004011938A2 (en) 2004-02-05
AU2003269041A1 (en) 2004-02-16
FR2842747A1 (en) 2004-01-30
FR2842747B1 (en) 2004-10-15

Similar Documents

Publication Publication Date Title
WO2004011938B1 (en) Method and device for screening molecules in cells
US9968903B2 (en) Apparatus for assay, synthesis and storage, and methods of manufacture, use, and manipulation thereof
EP3402890B1 (en) Intracellular delivery of complexes
JP4361271B2 (en) Instruments for assay, synthesis, and storage, and methods of making, using, and operating the same
US7442339B2 (en) Microfluidic apparatus, Raman spectroscopy systems, and methods for performing molecular reactions
Groll et al. Ultrathin coatings from isocyanate terminated star PEG prepolymers: patterning of proteins on the layers
JP2009504161A (en) Apparatus for assay, synthesis and storage, and methods for making, using and operating the same
JP2009504161A6 (en) Apparatus for assay, synthesis and storage, and methods for making, using and operating the same
Roy et al. Engineered cell culture substrates for axon guidance studies: moving beyond proof of concept
JP2010513902A (en) Sample Analyzer All documents cited herein are incorporated by reference in their entirety.
JP2002541458A (en) Miniaturized cell array method and apparatus for cell-based screening
FR2823999A1 (en) MINIATURE DEVICE FOR SEPARATING AND ISOLATING BIOLOGICAL OBJECTS AND USES
EP0842411A1 (en) Apparatus for the parallel alignment of macromolecules, and use thereof
US7659053B2 (en) Methods of alteration of surface affinities using non-chemical force-creating fields
EP1644731B1 (en) Method and device for the analysis of living reaction media
US7005294B2 (en) Method for producing an array for detecting constituents from a biological sample
US20080248972A1 (en) Method of Immobilizing Protein, Protein Chip, Method of Immobilizing Cell and Cell Chip
US20080113352A1 (en) Electrochemical Patterning on Multi-Channel Microelectrode Array for Biosensing Applications
Hager et al. Tunable DNA Hybridization Enables Spatially and Temporally Controlled Surface-Anchoring of Biomolecular Cargo
Flesch et al. Three-dimensional interfacing of cells with hierarchical silicon nano/microstructures for midinfrared interrogation of in situ captured proteins
CN1599640A (en) Biopolymer synthesis substrate and method for producing biopolymers
KR100473815B1 (en) Method and device for electronic control of the spatial location of charged molecules
Meza Stratégies de fonctionnalisation pour le développement de biopuces innovantes
KR20230171970A (en) How to Fabricate Composite Arrays
WO2014026271A1 (en) Surface chemical gradients

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
B Later publication of amended claims

Effective date: 20040806

WWE Wipo information: entry into national phase

Ref document number: 2492933

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2004523860

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 2003750828

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 2003750828

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 2003750828

Country of ref document: EP