EP1796843B1 - Vorrichtung zur dielektrophoretischen trennung von partikeln in einer flüssigkeit - Google Patents

Vorrichtung zur dielektrophoretischen trennung von partikeln in einer flüssigkeit Download PDF

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Publication number
EP1796843B1
EP1796843B1 EP05800525A EP05800525A EP1796843B1 EP 1796843 B1 EP1796843 B1 EP 1796843B1 EP 05800525 A EP05800525 A EP 05800525A EP 05800525 A EP05800525 A EP 05800525A EP 1796843 B1 EP1796843 B1 EP 1796843B1
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Prior art keywords
electrodes
fluid
potential
particles contained
sets
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French (fr)
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EP1796843A1 (de
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Pascale Pham
François PERRAUT
Adrien Plecis
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Commissariat a lEnergie Atomique et aux Energies Alternatives CEA
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Commissariat a lEnergie Atomique CEA
Commissariat a lEnergie Atomique et aux Energies Alternatives CEA
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C5/00—Separating dispersed particles from liquids by electrostatic effect
    • B03C5/02—Separators
    • B03C5/022—Non-uniform field separators
    • B03C5/026—Non-uniform field separators using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]

Definitions

  • the invention relates to a device for performing the dielectrophoretic separation of a fluid, and in particular a liquid, in particular to allow the isolation or collection of particles in the broad sense, contained in such a fluid.
  • these particles consist, without limitation, of biological cells, such as bacteria (a few tens of micrometers) and / or biomolecules (DNA, enzymes, proteins, liposomes ...), whose sizes can go down to a few tens of nanometers, even a few nanometers.
  • these objects can consist of molecules, or aggregates of molecules (micelles).
  • these objects may consist of solid particles in a liquid medium (suspension), colloids or even aerosols.
  • biosafety we can mention the field of biosafety, sanitary controls, agrifood quality controls, the search for new drugs. It is also possible to mention applications using micro-capsules and micro-spheres (paints, cosmetics, food industry), aerosols (atmospheric pollution), etc.
  • the particles subjected to the electric field gradient do not "see” the change of sign of the applied electric field. In doing so, it is possible to move a polarizable particle by dielectrophoresis with an alternating signal.
  • the implementation of the alternating electric field makes it possible to reduce or even eliminate the parasitic electrochemical reactions that may occur in particular at the level of the electrodes in the electrical systems in ionic liquid solution. We try to fight against these phenomena, insofar as they generally induce gaseous releases to the electrodes, and also modify locally the chemical characteristics of the media.
  • electrodes generating an electric field gradient are deposited on a flat surface (glass, passivated silicon, etc.) thus leading to planar configuration systems.
  • the fluid and the particles contained therein are in contact with the upper plane of the electrodes.
  • FIG. 1 a cross section of a planar configuration with interdigital electrodes.
  • Planar configurations however, have a number of major disadvantages, which will be described below.
  • the dielectrophoretic force F DEP has a small range in the direction perpendicular to the plane of the electrodes, that is to say in the volume of the fluid containing the particles (axis oz in the figures).
  • the force is maximal in contact with the edge of the electrode.
  • the edge of the electrode creates a wedge effect, at which the electric field is maximum. It is further demonstrated that the range of the dielectrophoretic force according to oz is effective in a zone of radius equal to about 40% of the parameter d, that is to say the distance between the center of the inter-electrode gap and the center of the electrode in question.
  • the collection of particles under the effect of dielectrophoretic forces is effective in volume, if the dimension h of the fluid located above the electrodes is of the order of magnitude of the pattern d of the electrodes. In other words, this efficiency is more limited, or requires working with very limited volumes of the fluid to be treated.
  • planar configuration systems Another major disadvantage of planar configuration systems lies in the fact that the electrical nature of the particle-fluid pair can make collection ineffective due to a negative dielectrophoresis regime.
  • the direction of the dielectrophoretic force developed by the planar electrodes depends, on the one hand, on the frequency of the electrical signal applied to the electrodes, but also on parameters independent of the actual power supply, namely the electrical properties of the particle pair. /fluid.
  • the influence of the value of the electrical conductivity of the carrier fluid of the particles on the dielectrophoresis regime is particularly significant.
  • a component designed to collect particles by dielectrophoretic attraction is inefficient if the electrical conditions, and in particular the nature of the particle - fluid pair, make the dielectrophoresis regime always negative.
  • a too conductive fluid can render a planar configuration component incapable of any collection on its electrodes.
  • this kind of problem is commonly encountered in biology, where the liquids are generally aqueous ionic solutions, therefore highly conductive.
  • the dielectrophoretic forces can be inhibited by concurrent forces also from the applied electric field, including electro-convection.
  • electro-convection we mean all the phenomena of setting in motion of the fluid (convection because of the existence of an electric field which is applied to it) and in particular the setting in movement by electro-osmosis (presence of charges on the electrodes) and Joule warm-up (presence of an electric current in the fluid).
  • the moving fluid causes the particles because of their small size: this convection movement is then superimposed on the dielectrophoretic movement, which can sometimes be completely inhibited if the accumulation zones associated with each phenomenon are not the same.
  • Electro-convection then constitutes a parasitic phenomenon, which is found in particular in systems with planar configuration, where the drive by electro-convection generally goes against the dielectrophoretic forces: for example in systems with interdigital electrodes, electro-convection induces the creation of accumulation zones located in the middle of the electrodes and / or in the center of the inter-electrode space, which are not located in the same place as those due to dielectrophoresis, constituted, as already said, by the edge of said electrodes.
  • This phenomenon of electro-convection is a phenomenon that depends on the power supply frequency of the electrodes, and which is all the more important that the particles are small.
  • this phenomenon decreases as the frequency increases, whereas the positive dielectrophoresis requires not to work above the cutoff frequency, corresponding to the frequency marking the change from positive dielectrophoresis regime to negative dielectrophoresis.
  • the object of the present invention therefore aims to separate particles from a fluid by dielectrophoresis, overcoming all of these various disadvantages.
  • the device according to the invention for the dielectrophoretic separation comprises two types of electrodes, each of the two types of electrodes being brought to a different potential, so as to generate an electric field within said fluid, both types of electrodes.
  • electrode being positioned within a chamber or pipe receiving the fluid subjected to dielectrophoretic separation, said enclosure itself being provided with a particle collecting surface.
  • the electrodes lose their role of collection surface and have only a limited electrical role, namely to deliver a non-uniform electric field, in order to produce effective dielectrophoretic forces for collection and directed to the collection surface, and thus to the bottom of the enclosure or the pipe.
  • the two types of electrodes are alternately supplied with electric current.
  • One of the objectives of the invention is to obtain, on the one hand, a dielectrophoretic force parallel to the oz axis, ie perpendicular to the collection plane, and on the other hand, distributed in a controlled manner according to ounces
  • the intensity of the di-electrophoretic force may be of substantially constant intensity along the axis oz.
  • the electrodes no longer constitute a collection surface of the particles to be separated, the dimensions of said electrodes therefore no longer constitute a limiting factor for the reading step. their size can be adapted to the volume of fluid to be treated.
  • the device can operate both in positive dielectrophoresis and in negative dielectrophoresis, thus making it possible to significantly increase the fields of application of the present invention.
  • the efficiency of the device of the invention is no longer dependent on the type of dielectrophoresis regime. It should be remembered in this respect that the aforementioned planar configurations necessarily require a positive dielectrophoresis regime, to perform the collection on a solid surface.
  • the potential V (z) will be decreasing with oz, and applicable to a determined particle-fluid set and with a signal frequency of electrodes also determined.
  • the signal V (z) is inverted with respect to the preceding configuration, in order to maintain a dielectrophoretic force always directed toward the collection surface, especially if the fluid becomes very conductive, or if wants to work with another frequency.
  • microelectronics techniques already used to produce the planar systems can be preserved for the realization of these electrodes. They can be assembled in a macrosystem which contains the collecting surface and which must perform all the other non-electrical functions (sealing, fluid supply, connection to a reading system, etc.) associated with the component according to its type of use. (capture, separation, sorting, etc ). They can also be performed in a microsystem.
  • the invention recommends, according to a first embodiment, called “beveled electrodes", according to the figure 5 , that the electrode groups A and B are each composed of a single electrode, supplied at the peak value potential V 0 , whose respective surface in contact with the fluid has an inclination of an angle ⁇ relative to the horizontal, giving them a beveled appearance.
  • the electrodes have a rectangular trapezoidal longitudinal section, whose inclined face is in contact with the fluid.
  • the angle ⁇ depends on the volume of fluid to be treated and the nature of the particle-fluid pair: it must satisfy the condition 0 ⁇ ⁇ ⁇ 90 °.
  • Beveled electrodes is equivalent to the configuration obtained with two facing electrodes, which are inclined at an angle ⁇ , always with respect to the horizontal illustrated in relation to the figure 6 .
  • the compensation of the transition from a positive dielectrophoresis regime to a negative dielectrophoresis regime can be done either by reversing the inclination of the electrodes ( figure 7b ), or by moving the collection surface C on the upper part of the component, as shown in FIG. Figure 7c .
  • a negative dielectrophoresis regime is implemented in the Figures 7b and 7c , respectively by inverting the profile of the electrodes, in order to result in a decreasing variation of the potential as a function of oz, and by positioning the collecting surface at the upper level of the storage or displacement chamber of the liquid to be treated and retaining the increasing variation of the potential with the axis oz.
  • the invention proposes a second embodiment called "isolated electrodes", more particularly described in relation to the figure 8 .
  • the electrode groups A and B are each composed of a single electrode, supplied at the peak value potential V 0 , each of said electrodes being coated at its face in contact with the fluid, with a layer made of an electrical insulating material I.
  • the deposition of this layer of insulating material is made such that the surface of said insulator in contact with the fluid has an inclination of an angle ⁇ relative to the horizontal. In other words, this amounts to varying the thickness of the insulation layer along the axis oz.
  • the invention consists in playing on the thickness of the insulating layer to create a variable potential V (z) along the electrode and along the axis oz.
  • V (z) variable potential
  • the actual electrode has a surface parallel to the direction oz and it is the variable thickness insulation with z that creates the non-constant function V (z).
  • the nature of the insulating material is not predefined. It must be chosen so that it ensures a good mechanical adhesion on the electrode, a good homogeneity to the impermeability of the electrical charges and mechanical properties which make it easily machinable.
  • the use of isolated electrodes can bring a very clear improvement in the performance of a dielectrophoresis system.
  • the presence of electric fields in the conductive fluids can induce electric charge transfers at the electrodes, thus capable of generating electrochemical reactions.
  • These electrochemical reactions to the electrodes are all limiting factors to the efficiency of the separation, because they generally cause gaseous releases that quickly degrade the electrical performance of the component.
  • the intensities of the applied electric fields are mainly limited by these electrochemical effects. However, if the intensity of the applied fields is increased, the intensity of the dielectrophoretic forces resulting therefrom are also increased, thus optimizing the effectiveness of the component.
  • the insulating layer prevents electrical charges from passing between the fluid and the electrode in question. It thereby limits the occurrence of electrochemical reactions to the electrodes and allows working with higher electric field levels (ie applied potential levels V 0 ) than those usually achieved with uninsulated electrodes.
  • the increase in the intensity of the electric field leads to more intense dielectrophoretic forces.
  • the performance of the devices employing such isolated electrodes is better, regardless of their geometric configuration.
  • each electrode group A and B consists of a stack of electrodes, fed by an electrical signal individually, and separated by an insulating material.
  • the number N of stacked electrodes in each group and their size according to oz are not fixed. Each group must have at least two electrodes and their increasing number N enhances the desired performance of the component.
  • the stacked electrode configuration can be used either by simultaneously applying to each of the two groups A and B of electrodes a different potential (V 1 , V 2 , V 3 ) on each electrode (spatial variation of the potential), or by applying a potential (constant or not) sequentially on each electrode (temporal variation of the potential).
  • the electrodes are consecutively "lit” one after the other, ie they are brought to the same potential consecutively, inducing a spatio-temporal gradient of the potential and a dielectrophoretic force which, in time, is moves towards the capture surface, conferring a piston effect on the particles.
  • each electrode of each group is indicated on the electrical diagrams represented in relation to the Figures 11a, 11b and 11c .
  • an impedance Z i composed of a combination of resistance and inductance R i L i , is placed across the terminals of each electrode.
  • a configuration without phase shift is obtained with the electrical diagram of the figure 11b , limitingly implements a resistance, and thus causing a spatial variation of the potential V.
  • the electrical diagram of the figure 11c using inductances, a spatio-temporal variation of the potential V is obtained, the inductance inducing a delay.
  • FIG. 12a and 12b illustrate a pyramidal checkerboard structure obtained from a beveled electrode configuration, respectively in cross section and seen from above.
  • the checkered structure component can be adapted to microwell plates already used for this type of application. These plates have microcuvettes, generally distributed in matrix. The flanks of the cuvettes may constitute the support of the electrodes implemented in accordance with the invention.
  • Each well consists of an elementary pyramidal component and acts as a pad capable of chemically differentiating, by the nature of the capture surface positioned at the bottom of the well, a desired molecule.
  • the individual ignition (addressing) of each pad consists in applying an electric potential on each group of electrodes. Ignition of the wells simultaneously or sequentially promotes the capture of molecules by dielectrophoresis.
  • the main interest of this particular configuration is to find the operation of a planar system while separating the electrical surfaces of the capture surfaces.
  • the collection is improved if an insulating base is used as the collection surface. Indeed, it is demonstrated that with such a collection surface, it avoids the concentration of particles collected at the electrodes, that is to say at the place where the electric field is the most intense.
  • the insulating base then acts as a stopping or confinement zone, which is no longer in contact with the electrodes.
  • this insulating base is replaced by a base made of a conductive material, electrically isolated from the electrodes, and carried for example to ground or polarized.
  • the substrate to be conductive it advantageously has a layer made of gold, silver, platinum, aluminum or chromium.
  • a layer made of gold, silver, platinum, aluminum or chromium To be more transparent, it can be made in ITO (generic term designating the oxides of Indium) or polyaniline.
  • the detection can thus be carried out optically, and in particular by fluorescence, whether the base is transparent or not. In the latter case, we go through the excitation of fluorescence via a surface plasmon. This detection can also be carried out in surface plasmon resonance. It can also be performed electrically then using the base as an active electrode during a read operation.
  • the device of the present invention is of interest inasmuch as, first and foremost, it makes it possible to define a field of dielectrophoretic forces extending within the entire volume of fluid, which the could not be obtained with the devices of the prior art.

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  • Engineering & Computer Science (AREA)
  • Microelectronics & Electronic Packaging (AREA)
  • Electrostatic Separation (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)

Claims (10)

  1. Vorrichtung zum Durchführen einer dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, zwei Elektrodenarten A und B umfassend, wobei jede der beiden Elektrodenarten auf ein anderes Potential gebracht wird, um in der Flüssigkeit ein elektrisches Feld zu erzeugen, wobei die beiden Elektrodenarten A und B in einem Behälter oder einer Rohrleitung positioniert sind, der bzw. die die der dielektrophoretischen Trennung unterzogene Flüssigkeit aufnimmt, wobei der Behälter selbst mit einer Partikelsammelfläche ausgestattet ist, dadurch gekennzeichnet, dass:
    - jede der beiden Elektrodenarten in die Flüssigkeit in dem Behälter oder der Rohrleitung eingetaucht ist und sich auf einer Ebene befindet, die sich von der Ebene der Partikelsammelfläche unterscheidet;
    - jede der beiden Elektrodenarten gegenphasig mit elektrischem Strom gespeist werden;
    - und das von den beiden Elektrodenarten erzeugte Potential einen Gradienten aufweist, der vom Abstand in der Richtung oz senkrecht zur Ebene der Partikelsammelfläche abhängt.
  2. Vorrichtung nach Anspruch 1, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass die Elektroden mit einer Schicht bedeckt sind, die aus einem elektrisch isolierenden Material hergestellt ist.
  3. Vorrichtung nach den Ansprüchen 1 und 2, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass die beiden Gruppen A und B jeweils aus einer einzigen Elektrode bestehen, die auf ein Potential mit einem Spitzenwert V0 gespeist werden, und dass die Oberfläche jeder der Elektroden, die mit der zu behandelnden Flüssigkeit in Kontakt ist, ein in Bezug auf die Waagerechte geneigtes Profil aufweist.
  4. Vorrichtung nach Anspruch 3, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass jede der Elektroden, die die Gruppen A und B bilden, einen rechtwinklig trapezförmigen Längsquerschnitt aufweist, dessen geneigte Fläche mit der Flüssigkeit in Kontakt ist.
  5. Vorrichtung nach Anspruch 3, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass jede der Elektroden, die die Gruppen A und B bilden, einen rechteckigen Längsquerschnitt aufweist.
  6. Vorrichtung nach Anspruch 3, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass jede der Elektroden eine mit der Flüssigkeit in Kontakt stehende Fläche senkrecht zur Ebene der Partikelsammelfläche aufweist, und dass die zur Flüssigkeit gerichtete Seite der Elektroden mit einer aus einem isolierenden Material hergestellten Schicht in einer zur Ebene der Partikelsammelfläche senkrechten Richtung zu- oder abnehmenden Dicke überzogen ist.
  7. Vorrichtung nach Anspruch 1, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass die beiden Gruppen A und B jeweils aus mehreren, in der zur Ebene der Partikelsammelfläche senkrechten Richtung übereinander gestapelten Elektroden bestehen, wobei die Elektroden in Zweiergruppen durch elektrisches Isoliermaterial getrennt sind, wobei die Elektroden jeder der Gruppen A und B einer räumlichen Veränderung in der Richtung oz des Potentials unterliegen, das an sie angelegt wird.
  8. Vorrichtung nach Anspruch 1, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass die beiden Gruppen A und B jeweils aus mehreren, in der zur Ebene der Partikelsammelfläche senkrechten Richtung übereinander gestapelten Elektroden bestehen, wobei die Elektroden in Zweiergruppen durch elektrisches Isoliermaterial getrennt sind, wobei die Elektroden jeder der Gruppen A und B einer sequenziellen und zeitlichen Veränderung eines konstanten oder nicht-konstanten Potentials unterliegen, das an sie angelegt wird.
  9. Vorrichtung nach Anspruch 8, zum Durchführen der dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass die Elektroden jeder der Gruppen A und B aufeinanderfolgend in der zur Ebene der Partikelsammelfläche senkrechten Richtung so auf ein bestimmtes Potential gebracht werden, dass ein räumlich-zeitlicher Gradient des Potentials und in der Folge eine dielektrophoretische Kraft induziert wird.
  10. Komplexe Vorrichtung zum Durchführen einer dielektrophoretischen Trennung von in einer Flüssigkeit vorhandenen Partikeln, dadurch gekennzeichnet, dass sie aus einem Verbund mehrerer Grundvorrichtungen nach einem der Ansprüche 1 bis 9 besteht.
EP05800525A 2004-10-04 2005-09-15 Vorrichtung zur dielektrophoretischen trennung von partikeln in einer flüssigkeit Active EP1796843B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR0410443A FR2876045B1 (fr) 2004-10-04 2004-10-04 Dispositif pour realiser la separation dielectrophoretique de particules contenues dans un fluide
PCT/FR2005/050745 WO2006037910A1 (fr) 2004-10-04 2005-09-15 Dispositif pour realiser la separation dielectrophoretique de particules contenues dans un fluide

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EP1796843A1 EP1796843A1 (de) 2007-06-20
EP1796843B1 true EP1796843B1 (de) 2011-08-17

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US (1) US8034226B2 (de)
EP (1) EP1796843B1 (de)
JP (1) JP4931822B2 (de)
AT (1) ATE520467T1 (de)
FR (1) FR2876045B1 (de)
WO (1) WO2006037910A1 (de)

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US20100018861A1 (en) * 2007-03-26 2010-01-28 The Regents Of The University Of California Electromotive liquid handling method and apparatus
US8246802B2 (en) * 2007-05-14 2012-08-21 The Regents Of The University Of California Small volume liquid manipulation, method, apparatus and process
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ATE520467T1 (de) 2011-09-15
JP2008516215A (ja) 2008-05-15
US20080011608A1 (en) 2008-01-17
FR2876045B1 (fr) 2006-11-10
EP1796843A1 (de) 2007-06-20
US8034226B2 (en) 2011-10-11
WO2006037910A1 (fr) 2006-04-13
JP4931822B2 (ja) 2012-05-16
FR2876045A1 (fr) 2006-04-07

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