EP2097160A1 - Microdevice for treating liquid specimens. - Google Patents
Microdevice for treating liquid specimens.Info
- Publication number
- EP2097160A1 EP2097160A1 EP07847689A EP07847689A EP2097160A1 EP 2097160 A1 EP2097160 A1 EP 2097160A1 EP 07847689 A EP07847689 A EP 07847689A EP 07847689 A EP07847689 A EP 07847689A EP 2097160 A1 EP2097160 A1 EP 2097160A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- electrodes
- drop
- liquid
- edges
- electrode
- 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.)
- Granted
Links
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5088—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above confining liquids at a location by surface tension, e.g. virtual wells on plates, wires
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/40—Mixing liquids with liquids; Emulsifying
- B01F23/41—Emulsifying
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/30—Micromixers
- B01F33/302—Micromixers the materials to be mixed flowing in the form of droplets
- B01F33/3021—Micromixers the materials to be mixed flowing in the form of droplets the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/30—Micromixers
- B01F33/3031—Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
- F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
- F04B19/00—Machines or pumps having pertinent characteristics not provided for in, or of interest apart from, groups F04B1/00 - F04B17/00
- F04B19/006—Micropumps
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
Definitions
- the invention relates to the field of the treatment of liquid samples, in particular by centrifugation or stirring of a drop of liquid.
- the proposed invention also relates to the field of discrete microfluidics, preferably used for continuous microfluidics (in channels) as soon as pumps, valves, walls are freed from the walls necessary for the confinement of the flow. ... etc.
- Discrete (or digital) micro-fluidics play an increasing role in the development of new micro-systems such as lab on chips, and many analysis steps can be performed in a chain using discrete micro-fluidics.
- Molecules of biological or medical interest are for example transported within drops that pass between various analysis steps such as biochemical functionalization, the injection of biomolecules by heterogeneous mixture.
- the proposed invention finds many applications in small scale mixing, small scale extraction, small scale separation or purification by centrifugation, concentration and then detection of biological targets, pumping in microfluidics, transmission of micro-fluidic movements, the rheological characterization of fluid samples in the form of liquid drops or gels.
- the invention also relates to the field of purification of biological samples, and extraction of biological constituents.
- chromatography is the most sensitive analytical technique currently available for assaying a substance in a biological sample.
- Electrophoresis allows selective separation of biological molecules based on their electrical charge. But the miniaturization of the electrophoresis remains delicate since the medium allowing the migration of the constituents to be analyzed is a very viscous gel. Inserting and then handling a gel in a lab on chips analysis chain is difficult to implement.
- centrifuges exploited in biology, biochemistry or in the medical diagnosis to isolate constituents or purify biological samples, they consist of a shaft carrying a special rotor, all driven by a powerful motor.
- the rotor has locations, symmetrically located on either side of the axis, which can receive small test tubes containing the biological preparations to be analyzed or purified. The whole is enclosed in a tank, sealed during rotation, for security reasons.
- the proposed invention is a solution to two problems posed by current centrifuges: - the imbalance of the rotor to compensate permanently, and the difficulty of miniaturization since the centrifugal acceleration is also proportional to the radius of gyration.
- the present invention uses the setting in motion of fluid in a drop, which is itself at rest.
- the proposed invention applies to liquid inclusions, not in motion as in electrowetting techniques, but at rest (in static position).
- a liquid inclusion is centered on an EHD chip ("electrohydrodynamic") also object of the invention.
- EHD chip electrohydrodynamic
- This allows to generate an intense and organized movement, or brewing, to the interior of the drop and optionally outside, in the external fluid to the drop, for example if it and the EHD chip are covered with a viscous fluid, the drop being in a static position and not deforming .
- there is no block displacement or interfacial deformation of the liquid inclusion A movement, or displacement, before or after the brewing operation can take place, to bring the drop or the liquid inclusion in the brewing place or to move away after brewing.
- Viscosities are a kind of relay of the interfacial tangential impulse.
- the centrifugation according to the invention thus allows microfluidic miniaturization.
- R measures the centrifugation relative to the gravity or gravity, u ⁇ being the centrifugation speed) to be achieved: at first sight, more wide length of the liquid sample is small (case of micro-systems), the more it seems difficult to achieve significant centrifugation intensities.
- the present invention makes it possible to overcome this difficulty and retains most of the advantages associated with centrifugation as an analysis technique, in particular a biological one, while allowing its miniaturization and the associated advantages:
- a device is a device for forming at least one circulating flow, or vortex, on the surface of a drop of liquid, comprising at least two first electrodes forming a plane and having edges in view one on the other hand, such that the line of contact of a drop, deposited on the device and fixed with respect thereto, has a tangent projecting in the plane of the electrodes an angle strictly comprised between 0 ° and 90 ° with the edges facing each other of the electrodes.
- the shape of the electrodes makes it possible to promote the existence of fluids, the contours facing the electrodes being neither totally tangent nor totally perpendicular to the triple line.
- tangential interfacial movement is induced by the electric field, despite the smallness of the liquid sample, by the application of a tangential electric stress at the interface of a liquid sample, in the zones situated above the electrode interface areas.
- the angle, strictly between 0 ° and 90 °, between the tangent to the triple line (or its projection) and the edges facing each other of the electrodes, may advantageously be between 40 ° and 50 ° for example equal to substantially 45 °.
- the edges of the electrodes facing each other can be for example in the form of zig-zag or logarithmic spiral form.
- the electrodes are for example 2, 4, or 8.
- edges of the electrodes making, with the projection of the line of contact, an angle strictly between 0 ° and 90 °, alternate with edges of electrodes making an angle of 90 ° with this same projection.
- Means may be provided to activate and deactivate, successively, the electrodes. According to a particular embodiment, this activation and deactivation successively in time takes place at high frequency, greater than 100 Hz.
- Space separation of the edges of the electrodes facing each other can be alternately (by traversing the electrodes in their plane, in the direction of clockwise or in the opposite direction) a first value and a second value, less than the first.
- a second set of electrodes may be located opposite, parallel to the first electrodes.
- this second set of electrodes also forms a device according to the invention.
- a device according to the invention may further comprise a counter-electrode shaped tip.
- the invention also makes it possible to produce a pumping device comprising at least one device according to the invention, as described above, and means for bringing a second fluid into contact with a drop of liquid disposed on the device.
- a pumping device comprising at least one device according to the invention, as described above, and means for bringing a second fluid into contact with a drop of liquid disposed on the device.
- Such a device may comprise a plurality of devices according to the invention.
- the invention thus makes it possible to micro-pump secondary flows or to accelerate micro-fluidic flows by setting up one (or more) micro-gear (s) consisting of one (or more) liquid inclusion (s) surrounded by a secondary and continuous liquid phase.
- micro-gear s
- the present invention is distinguished by the use of a fluid interface which causes a tangential movement of interfacial origin. The flow rate thus obtained is much higher than most current micropumps and accidental physicochemical contamination due to the presence of walls is avoided.
- the proposed invention also makes it possible to produce apparatus such as a mini-stirrer, or an analytical mini-centrifuge, or a mini-emulsifier, or a micro-centrifuge, or a mini-rheometer.
- a mini-rheometer measures viscosity and elasticity by measuring or visualizing flow velocity fields.
- the fluid in the proposed invention, is a viscous shear of interfacial and dielectric origin, in the proposed invention, a flow can be pumped whether or not there are thermal, chemical or ionic gradients,
- the proposed invention also has the following advantages: - a non-destructive and isothermal character: the involved liquid inclusion can therefore contain fragile constituents, which can be denatured with temperature or under the effect of ionic forces,
- the capacity to generate, within a liquid inclusion of typically millimeter size, an intense rotational or stirring movement is of the order of 10 or 100,
- the chip as well as the pulling techniques applied to the apex of the liquid inclusion proposed in the invention allow the specific selection of constituents after microfluidic concentration for the purpose of extraction, analysis or analysis. a posteriori detection.
- the invention also relates to a method of forming at least one circulating flow or vortex in a drop of liquid in a surrounding medium, having different dielectric properties and / or different resistivities with respect to each other, comprising the following steps:
- the applied field is oblique with respect to the liquid droplet - surrounding environment.
- the volume of the drop may vary with time.
- One or more circulating flows or one or more vortices may be generated in the drop.
- the invention also relates to a process for microfluidic concentration by mixing or centrifuging a drop of liquid, in particular for detecting antibodies, or antigens, or proteins or protein complexes, or DNA or
- RNA comprising carrying out a method of forming at least one circulating or vortex flow in said drop of liquid according to a method according to the invention.
- a detection step can be performed, after mixing or centrifugation, without displacement of the drop.
- a liquid extraction stage of the drop may moreover be provided. Then it is possible to transfer the extracted liquid to a detection zone.
- the extraction step may be carried out by electrowetting or by emission of droplets from a Taylor cone.
- the invention also relates to the formation of a microemulsion comprising:
- a method of pumping a secondary fluid, according to the invention, with a drop of a primary fluid comprises the implementation of a method of forming at least one circulating or vortex flow in said drop of primary fluid according to a method as described above, and the pumping of the secondary fluid by contact with the primary fluid, the forces present at the primary fluid interface - secondary fluid for driving the secondary fluid.
- An analyte extraction method of a drop of liquid according to the invention comprises:
- a deactivation of the (at least) first two electrodes and the formation of a capillary bridge between the first insulating surface and a wall comprising at least one other electrode, the electrical activation of the first electrodes and of the other electrode, and the breaking of the capillary bridge.
- a particle extraction method comprises the implementation of a method according to the invention as described above, the surrounding medium consisting of a second liquid containing particles which have previously sedimented on the surface. interface of the two liquids, then separating, for example by electrowetting, the lateral parts, containing the particles, and a central part of the drop.
- FIGS. 1A and 1B show a geometry of the EHD system in the case of electrodes activated by an alternating electric potential difference.
- FIG. 2 represents a segmented edge two-electrode EHD chip.
- FIGS. 3 and 5 each represent a four-electrode EHD chip with segmented boundaries.
- FIG. 4 represents an EHD chip with two electrodes with segmented boundaries.
- - Figure 6 shows a drop of water placed on an EHD chip with two electrodes segmented at ⁇ 45 °.
- FIGS. 7 to 9 each represent an electrode EHD chip whose internal boundaries are logarithmic spirals.
- - Figures 10 and 11 each represent an electrode chip EHD whose internal boundaries are either straight segments or logarithmic spirals.
- FIGS. 12A to 12C represent vertical extraction steps using a method according to the invention.
- FIGS. 15A to 15D show extraction steps of another method according to the invention.
- FIG. 16A and 16B each represent a device according to the invention, provided with trapping pads.
- the invention can in particular implement crosslinked liquid inclusions whose size may for example vary between 10 microns and centimeter.
- a liquid inclusion 12 is in a static position, placed symmetrically astride two electrodes 4, 6 (or more, in even or odd numbers), which can be brought to different, continuous or alternating electrical potentials ( figures IA, IB). These are, for example, electric potentials of the same absolute value but of opposite signs.
- These electrodes rest on a substrate 3.
- the drop may be separated from the electrodes by an insulating layer 10 and possibly by a hydrophobic layer 8. But the device can also function according to the invention without these layers 8, 10 , continuously or alternatively.
- the liquid contact line 20 - layer 8 (or layer 10) - ambient medium 22 is called a triple line.
- This line of contact in the form of a circle (but not necessarily), does not deform, which is an important contribution, as regards the performance of stirring or centrifugation.
- Means 11 make it possible to apply between the two electrodes 4, 6 a difference in potential which gives rise to an oblique electric field with respect to the liquid 12 / liquid 22 or liquid 12 / gas interface 22.
- This oblique field that is to say, neither totally tangent nor totally normal to the surface of the liquid inclusion 12, will allow an accumulation of electrical charges at the interface, and the creation of the momentum tangentially at the interface 12/22, amount of movement which, in turn, will cause currents 13, 15 internal to the drop, but no displacement of the drop itself.
- These currents appear in the plane of FIG. 1A for the sake of clarity, but rather they are oriented in a plane parallel to the plane of electrodes 4, 6 or layers 8, 10.
- An EHD chip according to the invention allows a mixture or a centrifugation not via the physical displacement of a drop by electrowetting but by the emergence of movements 13, 15 in the internal fluid to the drop and, optionally, in the external fluid to gout. These movements are generated by a viscous friction tangential to the surface of the considered inclusion.
- the invention thus makes it possible to produce, within liquid inclusions 12, using electrohydrodynamics (EHD), a micro-flow 13, 15 or a drainage, or a mixture (or mixing) of controlled intensity, or centrifugation.
- EHD electrohydrodynamics
- the nature, the thickness, the technological implementation of the layers 8, 10 are for example similar to those of the EWOD technology, such as for example described in the article by Y.Fouillet et al. cited above or in WO 2006/005880 or FR 2 841 063.
- the invention operates with various fluid couples 12/22 such as water / air, water / oil, water / chloroform ....
- the ambient medium 22 is preferably rather insulating (air, oil ).
- the drop 12 and the ambient medium 22 have different dielectric and resistive properties: different dielectric permittivities and / or different electrical conductivities; by way of example, mention may be made of the water / air or water / oil pairs, whose properties of dielectric permittivity and / or of electrical conductivity have the desired differences. For example with the water / oil pair or the water / air pair, the permittivity and conductivity jump is fully sufficient because the water is very strongly polarized (relative permittivity 80).
- the drop spreads and its shape does not change anymore.
- This voltage may for example vary from 0.1 V to 100 V or a few hundred V, for example 500 V.
- electrowetting the drop is kept centered or astride above the various electrodes. It is thus possible to use holding studs, as explained below.
- the normal component at the interface (also called the normal momentum balance) contributes to stably positioning the inclusion.
- the stirring or centrifugation results in particular from the tangential components of the previous equilibrium (tangential momentum balances) and more particularly from the tangential component along the tangent ti to the line of contact 20 of the liquid inclusion 12 concerned.
- the nature and the intensity of the mixture resulting from the internal currents 13, 15 can be controlled by controlling the vorticity level, the number and the size of the micro- or mini-vortex (s) generated within of the liquid inclusion.
- the geometry of the drop of water 12 is close to a truncated sphere, the normal n is oriented along the radial coordinate r, the tangents t 1 and t 2 are oriented along the longitude ⁇ and the co-latitude ⁇ , respectively.
- the dielectric permittivity ⁇ water and the dynamic viscosity ⁇ water in the drop of water 12 are much greater than their equivalents in the air 22 around the drop.
- the latter are separated from each other by an electrically insulating contour 16 in the form of a zig-zag: the segments are alternated at approximately 45 ° for a drop of water, as illustrated. in figures IB, 2 or 3.
- the periodicity (spatial) of the alternation, ⁇ can be optimized: it is preferable to take:
- R radius of the drop (3)
- R can vary between, for example, 0.1 mm and 10 mm.
- ⁇ can therefore be between, for example, 0.01 mm and 1 mm.
- ⁇ be the angle formed between the normal to the triple line 20 (contained in the so-called wetting plane), or its projection on the plane of the electrodes, and the edges 14, 16 of the electrodes.
- the absolute value of ⁇ is strictly between 0 ° and 90 °. An optimum configuration corresponds to an angle close to 45 °.
- this angle constraint is compatible with electrode edges having shapes such as, for example, zig-zag, or spiral.
- An envelope calculation makes it possible to take into account the angular stress ⁇ and leads to boundaries 14, 16 of electrodes in the form of a logarithmic spiral (or equiangular spiral).
- the median line separating the electrodes in their plane, or in the plane of the EHD chip, is described in polar coordinates by: where the symbol a is a homothetic factor.
- FIG. 1B shows a point M of polar coordinates p and ⁇ in a plane parallel to the plane defined by the electrodes 4, 6.
- the drop is arranged astride the electrodes. Locally, that is to say for two adjacent electrodes it is disposed on both sides of a direction ⁇ around which the electrode edges (zig-zag or spiral) oscillate, or which represents an average position electrode edges (see the direction ⁇ in Figures IB, 2, 7, but also the directions ⁇ and ⁇ 'in Figure 3).
- a possible instability of the static position of the liquid inclusion 12 can be countered by means of a sufficiently fast rotating electric field (at more than 100 Hz), obtained by the successive activations and deactivations of the electrodes 4, 6 with which the sample interacts: indeed, the liquid sample is then subjected to a motor electrical stress which sweeps its periphery
- the invention can be used for a stable volume 12, but also in the following different situations:
- the liquid inclusions 12, which are the object of mixing or centrifugation, have a non-constant volume (diameters changing from 100 ⁇ m to 10 mm),
- the drop 12 shrinks, or increases, under the effect of a phase change (interfacial mass transfer: evaporation / liquefaction), - after centrifugation, it may be useful to take a volume fraction of the liquid sample for purify it (extraction of a pellet or a supernatant), to extract constituents chemical or analytes ... etc. In this case, there is retraction of the drop after extraction.
- a phase change internal mass transfer: evaporation / liquefaction
- - after centrifugation it may be useful to take a volume fraction of the liquid sample for purify it (extraction of a pellet or a supernatant), to extract constituents chemical or analytes ... etc. In this case, there is retraction of the drop after extraction.
- the invention therefore remains effective if the volume of the liquid sample 12 is random or if it changes over time as a result of one or more extractions or under the effect of
- the invention allows easy integration within a lab-on-a-chip or a micro-system based on the displacement of liquid inclusions. Extraction techniques are proposed in the invention, which may for example implement means for moving drops by electrowetting, EWOD type, such as for example described in WO 2006/005880 or in the article by MG Pollack et al. . "Electrowetting based actuation of droplets for integrated microfluidics", Lab Chip, 2002, vol.2, p. 96-101.
- An interelectrode space e equal to 20 ⁇ m can be considered.
- the potential difference between two electrodes 4, 6 is typically set at 70V. If the surface of the liquid inclusion is sufficiently far from the inter-electrode space (thickness of the coating 8, 10 very large in front of e), the electric field lines emitted by two closely spaced electrodes adopt an axisymmetric geometry, and:
- E (p) -, (6) ⁇ p
- p denotes the distance between the median axis of the inter-electrode space and any point on the surface of the drop.
- R defined above generated with two electrodes can vary between 1 for a viscous gel and 100 for water. This is particularly the case for a liquid sample that has a relative dielectric permittivity equivalent to that of water (high).
- a first control parameter is the number of electrodes. With two electrodes 4, 6 vis-à-vis
- the number of electrodes can be increased in order to produce a cascade of recirculations and thus to control a mixture that is all the more rapid and efficient, especially if it is a question of mixing chemical or biochemical reagents.
- the increase in the number of electrodes leads to an increase in the number of inter-electrode spaces and therefore in the number of zones in which an oblique field occurs, motor of the mixing in the drop.
- a second control parameter is the angle between the nip and the boundaries of the electrodes.
- a second possibility is based on another control parameter, the inter-electrode spacing.
- the inter-electrode spacing To obtain a net non-zero balance of all the electrical motor constraints imposed around the drop on its surface one can impose, once on two, a wider inter-electrode spacing, typically by a factor of 10, than the preceding or the following, as described below, in connection with FIG. 9.
- the motor stress changes as the square of the imposed electric field which itself is proportional to the imposed potential difference and inversely proportional to the distance e between the electrodes buried under the insulating film, and inversely proportional to the thickness of the dielectric and hydrophobic films 8, 10.
- the electrode boundaries are represented, in top view, in the form of a zig-zag at 45 ° (see in particular FIG. 2 and the triple line 20 '') with the tangent to the triple line 20 of gout.
- the dashed circles 20, 20 ', 20' ' represent the triple line 20 which delimits the wetting area between the liquid sample and the surface of the EHD chip. They illustrate the possible variability of the liquid sample volumes 12, at various times t, t + dt, t + n. dt
- FIG. 2 is an example of an EHD chip according to the invention, with two electrodes 4, 6 with segmented boundaries
- FIG. 3 is an example of an EHD chip according to the invention, with four electrodes 4, 6, 24, 26 with segmented boundaries.
- the circle (thick line) delimits the contact line 20 of the liquid sample 12.
- the symbols E, E t and q s respectively denote the electric field in the inter-electrode space, the component from this field tangential to the triple line, and the electrical charge accumulated on the surface of the fluid sample under the effect of the normal jump of the electric field and the electrical characteristics (conductivity, dielectric permittivity).
- FIG. 4 is an example of an EHD chip according to the invention, with two electrodes 4, 6 with segmented boundaries. Two co-rotating vortices 13, 16 (dashed) are potentially generated.
- an EHD chip according to the invention has four electrodes 4, 6, 24, 26 with segmented boundaries. Four co-rotating vortices (dashed) are potentially generated.
- Isolated constituents can also be extracted within a vortex in view of their subsequent elimination, biochemical characterization or detection.
- FIGS. 7 and 8 show chips according to the invention, respectively with two or four electrodes 4, 6, 24, 26 optimized to take into account the volume variability of the liquid samples: the internal boundaries 30, 30 ', 32 ,, 32 'of the electrodes are logarithmic spirals.
- the line 20 of contact (dashed) is circular.
- the electric potentials (-) (+) are distinguished by their opposite signs: to two adjacent electrodes are applied opposite signs (except for an odd number of electrodes, for centrifugation, but this except for the rotating field).
- the EHD chip of FIG. 9 has eight electrodes optimized to: take into account the volume variability of the liquid samples: the internal boundaries 30, 30 ', 32, 32', 34, 34 ', 36, 36' of the electrodes are logarithmic spirals, and force the presence of a single vortex for the purpose of centrifugation.
- the thicker spirals 30 ', 32', 34 ', 36' signal an electrode boundary separation gap wider than the spirals 30, 32, 34, 36.
- the nip 20 (dashed) is circular.
- the electric potentials (-) and (+) are distinguished by the opposite signs of two neighboring electrodes.
- the electrodes delimited by the electrode boundaries are alternately a positive potential and a negative potential.
- the alternation of wider inter-electrode zones and less wide interelectrode zones makes it possible to significantly reduce, in the wider zones, the level of electrical stresses which otherwise would oppose the electrical motor stresses generated. by the smaller inter-electrode areas.
- the EHD chip has four electrodes 4, 6, 24, 26 and 8 electrodes 4, 6, 24, 26, 44, 46, 64 and 66 respectively, optimized for:
- the internal boundaries of the electrodes are alternately right segments and logarithmic spirals
- each electrode brought to a certain potential, may itself be cut locally in a circular contour (segmented electrode). This cutting makes it possible to create an artificial roughness facilitating the attachment of the line of contact of the drop.
- the portion of the electrode outside the contact line 20 can be deactivated, which can also stabilize the triple line by non-wetting.
- FIGS. 10 and 11 allow a trapping of the triple line, because of the circular cutting of the electrodes.
- Another interesting variant consists in stabilizing the position of the liquid sample by means of a difference in localized wettability at the level of the triple line.
- Figures 16A and 16B show studs 80, for example resin. They are preferably positioned furthest from the inter-electrode spaces, or in the inter-electrode spaces for which the component Et is to be suppressed; these are the larger inter - electrode spaces than their neighbors or the inter - electrode spaces that are locally orthogonal to the triple line.
- the pads 80 are made for example by photolithography of a thick resin layer (for example of thickness between 10 microns and 100 microns). In the case of FIG. 16A, the pads 80 make it possible to automatically center the drop in the center of the spiral.
- FIG. 16B they make it possible to automatically center the drop in the center of the spiral, and each is placed astride two electrodes where locally the electrohydodynamic stress is suppressed.
- a trapping of the triple line makes it possible to ensure the balance of the contact line 20 and to avoid any effect that may disturb the cohesion of the liquid sample 12 to be analyzed or treated. It also makes it possible to reinforce the stability of the static position of the drop 12.
- a chip according to the invention can be made with known technologies, for example as described in the document Fouillet et al., 2006, already cited in the introduction to this application or in WO 2006/005880 or FR 2 841 063.
- the drop is centered on the intersection of the inner edges of the electrodes
- the invention can be applied to extract analytes concentrated at the apex of a liquid inclusion 12 under the effect of centrifugal or centripetal forces.
- FIGS. 12a-12c show a three-stage extraction with two superposed horizontal walls: the lower horizontal wall is equipped with an EHD chip 2 according to the invention (according to one of the embodiments described in the present application) and the upper horizontal wall is equipped with an electrode 200, which is optionally an EHD chip according to the invention.
- the implementation steps are then as follows: i) centrifugation step on the lower horizontal wall equipped with the EHD chip 2 (FIG. 12a), by activation of this chip, and deactivation of the electrode of the upper wall.
- This first step makes it possible to promote the concentration of constituents at the apex (supernatant) or at the bottom, around the periphery of the liquid sample (pellet), depending on whether they are sensitive to centripetal or centrifugal forces, respectively.
- Taylor cone may also be useful for extracting isolated analytes at the apex of a liquid sample after brewing or centrifugation according to the invention.
- the liquid sample is placed on an EHD chip as proposed in the invention.
- a counter-shaped electrode is located in the opposing wall, as explained in the articles cited above in this paragraph.
- the operation can take place in three stages.
- the first step is to centrifuge the liquid sample to cause the microfluidic concentration of target constituents.
- the second step is to modify this action for a short time while carrying all the electrodes of the lower chip at the same potential while the upper electrode in the form of tip is brought to a very different potential.
- a capillary bridge is formed with the upper wall and in this case, the destabilization of the capillary bridge can be facilitated by activating a wider area of electrodes at the upper wall; it is therefore brought back to the previous technique, or there is ejection of one or more drops (electro-spray, as explained in the articles of Taylor, Ramos and Ganan-Calvo cited above).
- the constituents sediment and are concentrated in the form of a pellet in the residual bottom drop, or they supernatant and are then contained in the droplet or drops ejected by the Taylor cone. If these drops do not coalesce immediately (they have a similar electrical charge), their fusion can be facilitated later by electrowetting along the top wall.
- FIG. 13 represents a micro-pump implementing, for example, a four-electrode EHD chip (as for example in FIG. 10, but another number of electrodes is possible).
- a fluid inlet 72 makes it possible to introduce a secondary fluid 12 'into a cavity or a reactor 74 containing an EHD device according to the invention, here with 4 electrodes. Liquid inclusion primary 12 undergoes a treatment as already described above, without overall displacement. The surface forces in motion cause the secondary fluid 12 'by viscosity as described above, in accordance with the invention.
- a micro-pump according to the invention can be applied to a cooling process in microelectronics (for processors), or the dispensing of small quantities of drugs (pharmacology, galenic), or the micro-propulsion of objects (in exploration space).
- the speed range for mixing is considerably wider compared to conventional micropumps.
- the invention makes it possible in particular to reach a speed of at least 0.1 m / s or 1 m / s.
- the index i indicates that the quantity is evaluated at the interface, on the side of the primary fluid (p) or the secondary fluid (s).
- the drive of the secondary fluid is therefore more effective than its viscosity is low and yet higher than that of the primary fluid ( ⁇ p ⁇ s ).
- a first drop 12 it is also possible, from a first drop 12, to cause stirring or centrifugation in another drop by viscous drive even if the latter has a dielectric permittivity or electrical conductivity similar to those of the continuous liquid phase constitutive of the external medium.
- the ratio of reduction or amplification is programmable by adjusting the viscosity or diameter ratios between continuous liquid phase and drops.
- FIG. 14 shows a micro-fluidic gear involving, for example, two EHD chips 200, 202, preferably optimized (for example of the four-electrode type: FIG. 10), with their respective liquid inclusions 12, 112, one of characteristics: diameter dl and viscosity ⁇ l and the other of characteristics: diameter d3 and viscosity ⁇ 3. More EHD chips and liquid embedding can be implemented.
- a secondary liquid phase 212, viscosity ⁇ 2 circulates between the primary liquid inclusions 12, 112 through the movements of the latter, one in the direction of clockwise, the other in the opposite direction.
- the primary phase is for example a liquid sample placed on a chip according to the present invention.
- a movement of electrical origin is generated at the p / s interface which propagates within the secondary liquid via the viscosity. Therefore, at the s / t interface, there are two cases:
- a device of the micro-gear type according to the invention may comprise a series of inclusions, each resting on an EHD chip and connected to one another via the secondary liquid: in this case, such a micro-fluidic micro-gear amplifying the internal flows. and external to the inclusions is close to an amplification system.
- the secondary fluid and the fluid of each of the drops or inclusions have different dielectric permittivities and / or different electrical conductivities.
- this embodiment achieves a large number of G in one of the liquid inclusions participating in the chain ( Figure 14).
- the viscosity ratios of the fluids, the diameter ratios of the various inclusions involved, the number and the level of the electrical motor stresses applied to the various interfaces are all parameters that contribute to the overall amplification of the flows and that can be adjusted to optimize the flow. system.
- the present invention therefore makes it possible to generate a volume movement within a sufficiently viscous liquid sample via one or more electrical stresses exerted on its surface. If the liquid sample 12 is surrounded by another liquid 22, also viscous, the amount of movement induced by the surface electrical stress diffuse not only in the liquid internal to the liquid sample 12 but also in the external fluid 22. It is therefore possible to drive a secondary fluid in motion by means of a primary fluid adopting the form: either of one or more drops placed on one or more chips (FIGS. 13 or 14), or of a Capillary bridge trapped between two chips ( Figure 12a-12c),
- a micro-pump according to the invention may comprise a single inclusion of liquid embedded in a secondary fluid
- Figure 13 or several liquid inclusions embedded in a secondary fluid (Figure 14).
- the latter may be set in motion by a gear mechanism which may be referred to as an interfacial viscous friction micro-fluidic gear.
- Another embodiment of a process according to the invention comprises the steps of: centrifugation or microfluidic concentration,
- FIGS. 15A-15D A particular embodiment of this method is illustrated in FIGS. 15A-15D.
- the surrounding medium 22 consists of a second liquid, for example a second drop, immiscible with the first, containing particles 23. These particles 23 will gradually settle on the interface 12-22 (FIG. 15C).
- the setting in motion of this interface according to the invention, thus with the aid of electrodes having the characteristics already described above, without displacement of the drop 12, causes a displacement of the particles 23 along the interface 12-22 and their grouping on the edges of the drop 12.
- the lateral parts, containing the particles 23, are separated from the central part of the drop 22, for example by electrowetting cutoff, one or more of the electrodes situated between the one or more lateral parts and the central electrodes being deactivated. .
- the two drops are represented between, on the one hand, a substrate 3 on which is formed a device according to the invention and, on the other hand, a substrate 3 'of confinement.
- Microscale rheological instrumentation is an area of application of the invention.
- Micro-rheometers based on electrokinetics are currently in the development phase (Juang, Yi-I, 2006, Electrokinetics-based Micro Four-Roll MiIl 1 http: / / www.chambeng.ohio-s.ed.edu / facultypages / leeresearch / 154RollMill -hem).
- the proposed invention which is based on electrodynamics, makes it possible, for example, to generate four or two vortices within a liquid or gelled sample in order to obtain a purely elongational or purely sheared flow. Measurements of viscoelastic parameters can therefore be made with the invention using speed measurements made for example by video acquisition.
- a device according to the invention can be included in new micro-systems or on-chip laboratories, for the purpose of preparing biological samples before further analysis steps.
- PCR a process of amplification of the DNA strands present in a liquid sample.
- PCR is commonly developed in micro-systems (Kopp-MU, de-Mello-AJ, Manz-A, 1998, Chemical amplification: continuous-flow PCR on a chip, Science, 280, 5366, pp.1046-1048; Zhan-Z; Dafu-C-Y Zhongyao, Li W-Biochip for PCR amplification in silicon, 2000, st Annual International IEEE EMBS Special Topic Conference is Microtechnologies in Medicine and Biology Proceedings.. (Cat No.00EX451.) IEEE, Piscataway, NJ, USA, pp.
- PCR requires the preparation or purification of biological samples.
- the ELISA test is another widespread detection technique, such as immunoanalysis or viral load determination by nucleic acid assay, intended to detect and / or assay an antigen present in a fluid biological sample.
- the ELISA test practiced in homogeneous or heterogeneous phase, has the advantage of being fast and inexpensive.
- a first technique consists in hybridizing the target DNA segments with functionalized paramagnetic nanobeads responsible for vectorizing these segments to a functionalized solid interface for detection purposes.
- This concentration process can be based on a magnetic process, the target DNAs are eluted (by increasing the temperature above 50 ° C.) and hybridize on the functionalized solid surface before the detection phase (Marrazza, G ., Chianella, I. and Mascini, M., 1999, Disposable DNA Electrochemical Sensor for Hybridization Detection, Biosensors & Bioelectronics, 14, 1, pp.
- the present invention makes it possible to accelerate the hybridization kinetics while being compatible with a miniaturization constraint. It also makes it possible to concentrate by centrifugation the functionalized beads for a more sensitive detection. It is then applied as explained in document FR 01 11883.
- Another possibility consists in hybridizing target DNA strands at a liquid / gas or liquid / liquid interface functionalized by probes (Picard, C. & Davoust, L., 2005, Optical investigation of a wavy aging interface, Colloids & Surfaces A: Physichem Eng Aspects, 270-271, pp. 176-181, Picard, C. & Davoust, L., 2006, Dilational rheology of an air-water interface functionalized by biomolecules: the role of surface diffusion , Rheologica Acta, 45, pp.
- the present invention can be applied in two stages: it can be used to purify / prepare a liquid biological sample and then be used one last time by allowing a microfluidic type concentration.
- the invention makes it possible to locally concentrate ⁇ receptor-bound analyte ⁇ complexes in order to further increase the detection performance.
- An application of the invention therefore is especially the microfluidic concentration by mixing or centrifugation for facilitated detection of antibodies, antigens, protein or protein complexes, DNA or RNA.
- the fluids used are based on aqueous solutions.
- the ambient environment may be air or a pure oil.
- the detection can be carried out directly in situ at the level of the concentration zone or be the subject of a subsequent step after extraction by selective tearing of said concentration zone.
- the invention also makes it possible to improve the performance of PCR or PMCA for the detection of DNA or proteins.
- an EHD chip according to the invention can be optimized to take into account a variability of sample volumes (for example by a logarithmic spiral-shaped electrode chip, as illustrated in FIGS. 7-11).
- a microemulsion can also be achieved by promoting the coalescence of two electrowetting displacement inclusions and then producing a mixture using the present invention. PCR can then be performed directly on the emulsion thus obtained.
- the emulsion may also make it possible to eliminate certain unnecessary components by adsorption at the interfaces for biological purification.
- Two immiscible liquid inclusions can fuse with each other by the electrowetting technique, as described in the Y.Fouillet document already mentioned above.
- the invention then makes it possible to generate a two-phase mixture such as a foam or an emulsion (micro-foam, microemulsion), in order to facilitate sequencing, or the purification of biomolecules or even the extraction of colloids by capture at liquid / gas (foam) or liquid / liquid (emulsion) interfaces.
- a two-phase mixture such as a foam or an emulsion (micro-foam, microemulsion)
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FR0655327A FR2909293B1 (en) | 2006-12-05 | 2006-12-05 | MICRO-DEVICE FOR PROCESSING LIQUID SAMPLES |
PCT/EP2007/063178 WO2008068229A1 (en) | 2006-12-05 | 2007-12-03 | Microdevice for treating liquid specimens. |
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WO2008068229A1 (en) | 2008-06-12 |
JP5166436B2 (en) | 2013-03-21 |
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