The invention relates to electro-fluidic multiplexing for the manipulation of a plurality of drops in a microsystem.
The invention is particularly suitable for the lab-on-a-chip requiring the testing of a large number of different liquids, for example, for high-rate analysis or combinatorial chemistry.
The reaction volumes are drops manipulated by electrowetting on electrode series.
One of the most commonly used methods of movements or manipulation is based on the principle of electrowetting on a dielectric, as described in the article by M. G. Pollack, A. D. Shendorov, R. B. Fair, entitled “Electro-wetting-based actuation of droplets for integrated microfluidics”, Lab Chip 2 (1) (2002) 96-101.
The forces used for the movement are electrostatic forces.
Document FR 2 841 063 describes a device implementing a catenary opposite electrodes activated for the movement.
The principle of this type of movement is shown in
A drop 2 rests on an electrode array 4, from which it is isolated by a dielectric layer 6 and a hydrophobic layer 8 (
Each electrode is connected to a common electrode via a switch, or rather a system for individual control by electrical relay 11.
Initially, all of the electrodes as well as the counter electrode are placed at a reference potential V0.
When the electrode 4-1 located in the vicinity of the drop 2 is activated (placed at a potential V1 different from V0 by actuation of the relay 11), the dielectric layer 6 and the hydrophobic layer 8 between this activated electrode and the drop, polarised by the counter electrode 10, act as a capacitance, and the effects of the electrostatic charge cause the movement of the drop on the activated electrode. The counter electrode 10 can be a catenary as described in FR 2 841 063 (
The hydrophobic layer therefore becomes more hydrophilic locally.
The drop can thus be moved closer and closer (
The documents cited above provide examples of implementations of adjacent electrode series for the manipulation of a drop in a plane.
There are two families of production of this type of device.
In a first case, the drops rest on the surface of a substrate comprising the electrode array, as shown in
A second family of production consists of confining the drop between two substrates, as explained, for example, in the document of M. G. POLLAK et al. already cited above.
In the first case, it is an open system, and in the second case, it is a confined system.
The system generally consists of a chip and a control system.
The chips comprise electrodes, as described above.
The electrical control system comprises a set 11 of relays and an automatic system or a PC making it possible to program the switching of relays.
The chip is electrically connected to the control system, thus each relay makes it possible to control one or more electrodes.
Owing to the relays, all of the electrodes can be placed at a potential V0 or V1.
Generally, the number of electrical connections between the control system and the chip is equal to the number of relays.
To move a drop on an electrode line, it is simply necessary to connect all of the electrodes to relays and to activate them successively as described in
It is then desirable to simultaneously move (in parallel) N drops on these N lines.
For this, the electrodes are connected in columns, each electrode column being connected to a relay, called a parallel relay 20.
The operation of lines is dissociated in order, for example to bring a single given drop to one end, and to leave the other drops at the start of the line.
To dissociate the lines, at least one column of electrodes, called line selection electrodes, is defined, each of the electrodes of this column being connected, via a conductor 21-i, to a relay 22-i, which is independent of the relays to which the other electrodes of this same column are connected. These various relays are designated by the references 22-1, 22-6, 22-7, 22-8 in
All of the drops are moved on the N lines by parallel relays 20, up to the electrode column that precedes the column of line selection electrodes ESL.
By controlling the various line selection relays 22-i, it is possible to choose drops that are to be stopped and those that are to continue their movement along a given electrode line.
The drops thus selected can then continue their movement by the controlling of relays 20.
In this implementation, the number of electrical conductors 21-i and relays 22-i is proportional to the number of lines. For a large number of lines (N=20, 50, 100, etc.), the large number of conductors and relays makes this technology complex and very expensive.
Therefore, we have the problem of finding a method and a device making it possible to simplify the electrical connections while maintaining the possibility of selection for each line of electrodes.
The invention first relates to a device for addressing an electrode array of 2n lines of an electro-fluidic device, each line having N electrodes (n≦N), which device comprises:
The invention makes it possible to reduce the number of line selection conductors, and therefore to simplify the line selection means in an electro-fluidic addressing array.
Owing to the invention, it is therefore possible to manipulate 2n drops for only 2n input signals.
The invention therefore makes it possible to control line selection electrodes with only 2n relays.
For example, the invention makes it possible to control 8, 16, 32, 64, 128, 256, 512, 1024 line selection electrodes with respectively 6, 8, 10, 12, 14, 16, 18, 20 lines selection conductors and the same number of line selection relays.
The invention is particularly suitable when the number of lines is large (>16 or 32, for example).
The electrodes ESL-k for selecting the different lines can be, for a given value “k”, connected to two line selection conductors, the electrodes ESL-k being connected by packets of 2k−1 alternatively to conductor Ck and to conductor Ck′.
The selection means for selecting one or more line selection conductors can comprise electrical selection relays.
According to one embodiment, in such a device, the means for selecting line selection conductors comprise 2n electrical selection relays, each relay being connected to a single line selection conductor.
According to one embodiment, in such a device, the means for selecting line selection conductors comprise n electrical selection relays, each relay being connected to two line selection conductors.
Each line selection relay can then be combined with means for generating, in addition to an input signal, a complementary signal.
The line selection electrodes are arranged successively along each line, or non-successively along at least one line.
The line selection electrodes of at least one line can be in rectangular form, with the large side of each rectangle being arranged perpendicularly to the line.
The line selection electrodes of at least one line can be in square form according to an alternative.
According to a specific embodiment, at least one electrode line of the array has a cutting electrode (Ec).
Digital line selection means can be provided to control a device according to the invention.
These digital line selection means can be programmed to select the lines of the electrode array according to a binary code.
According to the invention, a combinatory logic is then used, which is obtained by a suitable method of interconnections between a plurality of electrodes at the level of the chip or of the device.
These digital line selection means can comprise means for selecting one or more lines of the array, and means for forming instructions for controlling line selection conductors according to the line(s) selected.
These digital line selection means can also comprise means for consecutively activating the line selection electrodes of a selected line and/or for simultaneously activating the line selection electrodes of a selected line.
The invention also relates to a device for forming liquid drops, comprising a device such as that described above, and means forming containers for liquids, each line of the array being connected to a container.
Such a device according to the invention can also comprise means forming 2n containers for liquids, each line of the array being connected to a single container.
Each line can be connected to a common line of electrodes, in order to mix the liquid drops formed on the different lines.
The invention also relates to a device for addressing an electrode array of p lines, with 2n<p<2n+1 lines, of an electro-fluidic device, comprising a device with 2n lines as described above.
The invention also relates to a method for moving at least one liquid volume, using a device as described above, comprising:
The line selection electrodes of said line can be activated consecutively or successively.
The invention also relates to a method for forming a liquid drop comprising the movement of a liquid volume as described above, the spreading of this volume on a plurality of electrodes of said line by simultaneous selection of these electrodes, and the cutting of the spread volume by means of a cutting electrode (Ec).
The implementation of the invention makes it possible to control a very large number of drops with simple chip production technology, a minimisation of the number of electrical connections between the chip and the control system, a simplification of the electrical control system, and therefore a minimisation of the costs of chip production, electrical connections and the control system.
One embodiment example of the invention will be provided in relation to
In this example, the device comprises 8 lines (No 0 to No 7) of electrodes, i.e. 23 lines.
Each line comprises at least 3 electrodes, with 6 in the example of
Among the electrodes of each line, 3 so-called selection electrodes Esl1, Esl2, Esl3 are selected. More generally, for N=2n lines, n selection electrodes Esl-i, i=1−n are selected on each line, n>0.
The line selection electrodes Esl-i are connected to line selection relays, as explained in greater detail below, or to line selection conductors C1, C1′, C2, C2′, C3, C3′ themselves connected to line selection relays.
In
In general, for N=2n lines, there are 2n line selection conductors.
The n line selection electrodes of each line, and therefore the 2n×n line selection electrodes, are connected to one or the other of the conductors of the n pairs of line selection conductors Ck, Ck′ (k=1, . . . n et k′=1, . . . n).
Each line selection conductor is controlled by a line selection relay, Rsl-k, Rsl-k′ (k=1-3, k′=1-3). Therefore, there are, in total, in this embodiment, 2n line selection relays.
The other electrodes, which are not line selection electrodes, are connected to parallel relays 30, as already explained above: each electrode column is connected to a parallel relay.
For a given line, the electrodes Esl-i are not necessarily consecutive: there can be, for at least one line, a “normal” electrode (which is not a selection electrode) between two selection electrodes Esl-i. Below, we will provide an example of the use of such a device.
In addition, it is preferable to adopt, by convention, a numbering direction common to all of the lines: for example, it is suitable for, on each line, the selection electrode the farthest to the right on the line to be Esl-1, with Esl-2 being the selection electrode to the left of Esl-1 (even if it is not juxtaposed with respect to it) and, more generally, with Esl-k being the selection electrode to the left of Esl-(k−1), even if it is not juxtaposed with respect to it.
For i=1, the electrodes Esl-1 of the different lines are connected to C1 and C1′ (then to Rsl-1 and to Rsl-1′) in an alternating manner: in other words, the electrodes Esl-1 are connected alternatively to C1 and C1′ (therefore, there is a change every 2(1-1) lines, i.e. at each line).
For i=2, the electrodes Esl-2 of the different lines are connected to C2 and C2′ (then to Rsl-2 and to Rsl-2′), again in an alternating manner, but every 2(2-1) lines, i.e. every two lines. In other words, groups of 21 electrodes Esl-2 are connected alternatively to C2 then to C2′.
For i=3, the electrodes Esl-3 of the different lines are connected to C3 and to C3′ (then to Rsl-3 and to Rsl-3′), again in an alternating manner, but every 2(3-1)=22 lines. In other words, groups of 22 electrodes Esl-3 are connected alternatively to C3 then to C3′.
More generally, for N=2n lines, 2k−1 electrodes Esl-k (k=1, . . . N) among all of the 2n×n electrodes Esl-k of all of the lines are connected to the line selection conductor Ck (connected to the relay Rsl=k the next 2k−1 electrodes being connected to the line selection conductor Ck′ (connected to the relay Rsl-k′). If there are more electrodes Esl-k after these two assignments, they may be assigned again to Ck (and therefore to Rsl-k) for the next 2k−1 electrodes, then again to Ck′ (therefore to Rsl-k′) for the next 2k−1 electrodes. If there is only one group of less than 2k−1 electrodes, they will be assigned either to Ck or to Ck′, depending on whether the previous electrodes Esl-k are connected to Ck′ or to Ck.
For a given value of “k”, the electrodes ESL-k of the different lines can be connected to two line selection conductors Ck or Ck′ (and to corresponding relays RSL-k or RSL-k′), the electrodes ESL-k being connected by packets of 2k−1, alternatively to conductor Ck and to conductor Ck′.
For a given line, the line selection electrodes of this line are assigned to different pairs Ck, Ck′ and therefore, in the configuration of
Finally, for the general case of 2n lines, 2n−1 line selection electrodes of 2n−1 lines are assigned or connected to each line selection conductor Ck.
In the case of
For example, for the line j=0, Esl-3 is activated if Rsl-3′ is also activated, and therefore also the conductor C3′ (
Regardless of the number of lines and line selection electrodes, each line selection conductor and each relay can have two different states.
A first state is called state “0”. The conductor Ck and the electrodes that this relay controls are then connected to the potential V0 (or to a floating potential): the electrowetting does not act on these electrodes. There is no movement or spreading of drops on these electrodes.
A second state is called state “1”. The conductors Ck and the electrodes that this relay controls are then connected to the potential V1: the electrowetting can act on these electrodes to move or spread the drops on these electrodes.
In order for a drop to cross line selection electrodes ESL1, ESL2 . . . , ESLn, of the same line, all of the line selection conductors and all of the relays to which these different electrodes are connected must be in state “1”.
If a single one of these line selection conductors or relays is in state “0”, there is no possible crossing of the liquid on the electrode lines connected to the line selection conductor and to the relay in state “0”.
If all conductors Ci and Ci′ and all relays RSLi and RSLi′ for i=1 to 2n are in state “0”, there is no possible crossing of liquid on any of the lines.
However, if all relays RSLi and RSLi′ are in state “1”, all of the drops can be moved or spread, on each line, on all electrodes ESL-1 to ESL-n.
This embodiment of the invention makes it possible to work with only 2n line selection conductors, and as many control relays, of the 2n×n line selection electrodes of all of the lines, with n line selection electrodes on each line.
On the contrary, the known devices implement, at best, 2n line selection electrodes, but with 2n conductors and as many relays (see
Relay control means 40 can also be provided, for example digital programmable means (PC or other) to which the relays are connected and which can control these relays.
These means can be equipped with a screen 42 enabling the user to select a line to which a drop must be capable of being transferred. For example, the array is shown on this screen, and the user selects one or more drop transfer lines, using a cursor or a pen enabling said user to designate the line(s) chosen directly on the screen.
Alternatively, an automatic program can select the lines and send corresponding control signals to the electrodes.
Means for storing means 40 make it possible to store the information enabling a given line to be selected. This information is, for example, that of table I in the case of an array for addressing 8 lines. It is stored or memorised in the form of table I or in another form.
Upon instruction by an operator, for example, upon a selection as described above, or upon an instruction of an automatic program, the digital means select, in the storage means, the data making it possible to open or close the necessary relays Rsl-k, Rsl-k′, and therefore to activate the necessary electrodes Ck, Ck′.
In the previous embodiment, the line selection conductors Ck, Ck′ are connected to as many line selection relays Rsl-k, Rsl-k′.
It is possible, according to another embodiment, to reduce this number of line selection relays.
Thus, according to another aspect of the invention, shown in
Each combination of n inputs of relays Rsl-k, and therefore a corresponding combination of line selection conductors Ck, Ck′, leads to the selection or to the opening of one or more lines of the array with a view to transferring a drop to this line.
For example, in the embodiment of
In this embodiment, there are only n relays.
It is also possible to encode or identify the 2n lines of the array by a binary code with n digits, each line being capable of being selected by assignment, to the input of n relays Rsl-k, of the coding for this line.
It is therefore possible in this case to implement a logic for encoding the lines as a binary number, and to assign this encoding to the line selection relay control, and therefore to the selection of lines themselves. To select a line, its binary code is assigned to the input of the line selection relays.
For example, reference can be made to 4, corresponding to the case of 8 electrode lines, comprising 3 line selection electrodes per line, 6 line selection conductors C1 to C6, but only 3 line selection relays.
In this example, the encoding of lines by using the state of the relays is summarised in table II below:
For a given binary digit, a single line will have the 3 line selection electrodes at potential V1, and a single line will be selected.
For example, the number 101 makes it possible to define the state of the 3 relays enabling the 3 electrodes ESL-1, ESL-2, ESL-3 of line 5 to be at potential V1.
Only the drops placed on this line can circulate.
The other drops cannot cross the electrodes ESL because at least one of them is at potential V0.
The assignments or the connections of the line selection electrodes to the line selection conductors Ck, Ck′ are, in this embodiment, the same as in the first embodiment.
Similarly, in this embodiment as well, relay control means 40 can be provided, for example, digital programmable means (PC or the like) to which the n relays are connected and which can control these relays.
These means can be equipped with a screen 42 enabling the user to select a line to which a drop must be capable of being transferred. For example, the array is shown on this screen, and the user selects a drop transfer lines, using a cursor or a pen enabling said user to designate the line(s) chosen directly on the screen.
Alternatively, an automatic program can select the lines and send corresponding control signals to the electrodes.
Storing means of means 40 make it possible to store the information enabling a given line to be selected, for example, the information of table II as provided above, in the form of this table or in an equivalent form.
Upon instruction by an operator, for example, upon a selection as described above, or upon an instruction of an automatic program, the digital means select, in the storage means, the data making it possible to open or close the necessary relays Rsl-k, and therefore to activate the necessary electrodes Ck.
In general, regardless of the embodiment envisaged, two modes of operation can be distinguished.
In a first case, for a given line, a drop is simultaneously spread on all of the line selection electrodes of this line; in a second case, the drop is moved successively over the line selection electrodes of this same line.
With the first mode of operation, the different line selection electrodes of the same line are simultaneously activated. For example, the control means 40 are programmed specifically in order to simultaneously activate these line selection electrodes. Or an operator can choose, on a case-by-case basis, between simultaneous activation and successive activation.
For this, the liquids and the technologies used (confined system or open system) enable the drops to be spread on the entire series of these line selection electrodes.
This is generally the case of confined systems. A confined system comprises, in addition to the substrate as shown in
For an open system, low surface tension liquids are preferably used (for example water with surfactants).
Depending on the surface tensions of the liquids and the sizes of the electrodes, it may be difficult to obtain a complete spreading of the liquid on all of the n line selection electrodes of the same line, activated simultaneously, when the number n is high (for example: n>3 or 4).
To overcome this problem, it is possible to modify the shape of the electrodes in order to minimise the total length of the different line selection electrodes, and therefore to limit the spreading length of the drop.
This is obtained, for example, by using rectangular line selection electrodes, as shown in
With the second mode of operation, the line selection electrodes are controlled consecutively.
Indeed, for some configurations, (for example, in an open system with high surface tension drops), it may be difficult to spread a drop simultaneously on all of the line selection electrodes of the same line.
By consecutively controlling the line selection electrodes of the same line (ESL-1 then ESL-2, up to ESP-n, or the reverse if the electrodes are numbered in the opposite direction), the drop selected is moved closer and closer along a line, on the different line selection electrodes placed consecutively at potential V1.
If one of the line selection electrodes is placed at potential V0, the drop is stopped.
To select a new drop, a resetting to zero is performed, which consists of replacing, at the start of the line, all of the drops stopped on one of the line selection electrodes. For example, the electrodes preceding the one on which the drop is located are reactivated, in order to cause the drop to move up along the line.
Alternatives for the formation of a drop will be described below.
It is possible to form drops from a container R by means of an electrode line that is connected to said container and that is itself part of an electrode array.
To this end, a series of electrodes E1 to E4 of a line of an array are activated, said line being connected to a container R as shown in
Then, the liquid segment obtained is cut by deactivating one of the activated electrodes (electrode Ec in
It is possible to apply the method according to the invention by inserting selection electrodes between the container R and one or more electrode(s) Ec (
According to the invention, the selection electrodes make it possible to select the lines where the drops must be formed, to spread the liquid up to the cutting electrodes in order to from a drop.
An example of an application will now be described in relation to
It relates to a fluid processor for combinatory chemistry.
In this example, the chip comprises 2×2n containers Rk, k=1, . . . , 2n+1, and a corresponding number of electrode lines.
Each container is associated with an electrode line making it possible to produce a drop. The lines together therefore form an array as already described above.
n line selection electrodes, as described above, are located on each line.
All of the electrode lines starting at the containers culminate in a common electrode line 60, which can also comprise line selection electrodes. The different reagents are brought to this line 60, in the form of drops, so as to be mixed.
The structure of 7A is symmetrical with respect to said line 60, and therefore comprises 2×2n lines. However, a structure according to the invention can also be asymmetrical and comprise only 2n lines, all located on the same side, or at 90° with respect to the common line 60.
The line selection conductors, arranged according to one of the embodiments of the invention, are not shown in
These line selection conductors are connected to control means such as means 40 and 42 of
According to an alternative, it is possible to have lines, each equipped with line selection electrodes and connected to a container R1, . . . Rk, R′1, . . . R′k, in a perpendicular architecture, according to an arrangement as shown in
According to yet another alternative, it is possible to have lines, each equipped with line selection electrodes and connected to a container R1, . . . Rk, R′1, . . . R′k′, R1, . . . Rj, R′1, . . . R′j′ in a square architecture, according t-o an arrangement as shown in
Other provisions can be envisaged and make it possible to produce any type of fluid processor or circuit.
The line selection conductors, arranged according to one of the embodiments of the invention, are not shown in
These line selection conductors are connected to control means such as means 40 and 42 of
Owing to the invention, it is possible to program a large number of possible combinations of mixtures between the various reagents.
To carry out the analysis of the results, the chip can comprise a detection zone (not shown in the figure) in which a detection can be performed, for example by colorimetry, fluorescence or electrochemistry.
The chip can optionally comprise other inlets/outlets or containers 62 for injecting a sample that is to be mixed, successively, with a combination of different reagents, each coming from a container connected to an electrode line, or to an area 64, called a waste receptacle area, for removing liquids after analysis.
The invention applies not only to arrays comprising 2n lines (n>0 or 1), but also to any array of p lines (p integer), with 2n<p<2n+1, n integer. In this case, an array of 2n+1 lines is processed according to one of the embodiments described above, then the excess lines in this pattern are suppressed.
The switches or relays are diagrammed by 4 blocks Rsl-i (i=1-4), which can take on one of the forms described above in association with one of the embodiments of the invention.
The suppression of, for example, 3 lines is symbolised by the dashed line 70. The lines j=13, 14, being eliminated, there is a configuration comprising 15 lines, including the 8 lines j=0-7, each of these 8 lines comprising at least 3 (in fact 4) line selection electrodes Esl-1, 2, 3, connected to the conductors C1, C1′, C2, C2′, C3, C3′ according to the invention (the block 72 of
The device also comprises two additional line selection conductors C4 and C4′, which, for lines 0 to 7, are respectively completely occupied or empty, and are not therefore involved in the identification of lines.
A device comprising p lines, with 2n<p<2n+1 therefore comprises a device with 2n lines according to the invention. Each of these lines no longer comprises n line selection electrodes, but n+1, of which n are connected as already described above in relation to
The invention therefore makes it possible to obtain a method and a system for addressing an electro-fluidic array having any number of lines.
A device according to the invention can be provided in a structure such as that shown in
The substrate 1 is, for example, made of silicon, glass or plastic.
Typically, the distance between the conductor 10 (
The conductor 10 is, for example, in the form of a wire with a diameter of between 10 μm and a few hundred μm, for example 200 μm. This wire can be a gold, aluminium or tungsten wire, or it can be made of other conductive materials.
When two substrates, 1, 11 are used, in the case of a confined structure (
In this case, the second substrate comprises a hydrophobic layer 13 at its surface intended to be in contact with the liquid of a drop. A counter electrode 15 can be buried in the second substrate, or a planar electrode can cover a large portion of the surface of the cap. A catenary can also be used.
Regardless of the embodiment considered, a liquid drop 2 will have a volume of between, for example, 1 nanolitre and several microlitres, for example between 1 nl and 5 μl.
In addition, each of the electrodes of a line of the array will have, for example, a surface on the order of a few dozen μm2 (for example 10 μm2), up to 1 mm2, according to the size of the drops to be transported, the spacing between neighbouring electrodes being, for example, between 1 μm and 10 μm.
The structuring of the electrodes of the array can be obtained by conventional micro-technological methods, for example by photolithography, the electrodes being, for example, produced by depositing a metal layer (AU, or AL, or ITO, or Pt, or Cr, or Cu), then photolithography.
The substrate is then covered with a dielectric layer of Si3N4 or SiO2. Finally, a hydrophobic layer can be deposited, for example Teflon using a spinner.
The same techniques apply to the production of the second substrate of
Methods for producing chips incorporating a device according to the invention can also be directly derived from the methods described in document FR 2 841 063.
Regardless of the embodiment, the electrodes of at least one line preferably have a saw tooth profile as shown in
An alternative embodiment of a device according to the invention will be described in relation to
This is an electrode array architecture, or a series arrangement of a plurality of multiplexes.
It is indeed possible to arrange a plurality of electrode systems in series as described above in relation to one of
Line selection electrodes Esl-i (i=1-3), Esl-i′ (i′=1-3), Esl-i″ (i″=1-3) are arranged on each line of electrodes. The number of 3 line selection electrodes is given by way of example and can be any number.
The other electrodes, which are not line selection electrodes, are connected to parallel relays 300, as already explained above: each electrode column is connected to a parallel relay.
The conductors Ci, Ci′ can be arranged as shown in
In this case, the electrodes Esl-1, Esl-1′, and Esl-1″ of the same line are activated at the same time. A drop, placed on one of the lines, will move closer and closer, from one electrode system to another arranged in series with it.
According to an alternative, not shown in the figures, each set of electrodes as described above in relation to one of
The series arrangement of a plurality of electrode systems, preferably comprising the same number of line selection electrodes, is applied not only to 3 electrode systems, each comprising 8 lines, as described above in relation to the example of
This type of series arrangement can also be applied to a device for addressing an electrode array of p lines, with 2n<p<2n+1 lines, of an electro-fluidic device, comprising a device with 2n lines according to the invention.
Another example of a chip according to the invention, making it possible to carry out storage and/or mixing and/or dilution operations, will be described in relation to
It comprises n containers (here n=16 by way of example; it is also possible to have any number n of containers, with n≧2) R1-R16 distributed in the following manner in the configuration shown:
The n containers communicate with one another (i.e. liquid volumes can be moved between these containers) by a bus 301 constituted by a line of electrodes. The drops are placed or dispensed on this bus 301 by way of the lines of line selection electrodes Esl-i, Esl-i′ in accordance with the invention. The control of these lines is, for example, one of the control modes described above in the context of this invention. The conductors Ck, Ck, as well as the relays Rsl are not shown in this figure for the sake of clarity. Various modes of operation of a container with one or more electrode lines were also described above in relation to
With this device of
An example of a mask layout used for the photolithography of the electrical level of the electrodes shown in
The bus 301 is constituted by a line of activated electrodes 3 to 3. Three relays make it possible to move a drop on the entire bus. The bus and its connection to the conductors 330, 331, 332 controlled by the relays is shown in greater detail in
References 320 and 321 of
Reference 400 designates another connection, from a line selection electrode 411 to a conductor 410 via a conductor 401.
A comb 340 groups all of the contacts. References 341 and 342 designate electrodes enabling contact at the level of a cover.
Not all of the line selection conductors are shown in this figure, for the sake of clarity.
Furthermore, conductive lines 343 come from the comb 340 in order to produce the connection of line selection conductors (shown or not) but also conductors performing other functions on the chip. In this case again, for the sake of clarity of the figure, the conductors 343 are not shown completely, but in an incomplete manner (they end in the figure in dotted lines).
In total, with a control system working with a limited number of relays, in this case only 16 relays, it is possible to control around one hundred electrodes in order to manipulate the liquids in the 16 containers. The number of relays is in fact dependent not only on the number of containers, but also on other functions to be activated on the chip.
The electrodes are formed by a conductive layer (for ex.: gold) with a thickness of 0.3 μm. The patterns of the electrodes and the connection lines are etched by conventional photolithography techniques. A deposition of an insulating layer is performed, for example with silicon nitride in a thickness of 0.3 μm. This layer is locally etched in order to be capable of taking up the electrical contacts.
For the second electrical level mentioned above, the technology used is the same as that for the electrode level, i.e. a metal deposition and photolithography. The interconnections (some mutual only) are designated by reference 400 in
For example, the chip is made of silicon and measures 4 to 5 cm2. The surface of each electrode of the bus 301 and the electrodes of containers R2 to R15 is 1.4 mm2. The surface of each selection electrode ESL is 0.24 mm2.
In one or more containers, and in particular in containers R1 and R16, the liquid can be moved by electrowetting toward the outlet of the container, i.e. toward one of the electrodes of the electrode line connected to said container.
In particular, in
In
According to an alternative, shown in
This improvement also makes it possible to completely empty the container.
It should be noted that the fingers of the comb (
Reference 470 designates an interconnection of the electrowetting electrodes between different lines.
Reference 471 designates an electrode of the comb 340 (
A thick resin (100 μm of thickness, for example) is rolled, then structured by photolithography, and a hydrophobic treatment is carried out (for example, Teflon AF by Dupont). This resin film is used to define the spacing 350, 351 between the lower plate 1 and the upper plate 11 (
An example of the operation of this device, or fluidic protocol, will be provided below.
With the chip described above, it is possible to carry out a protocol making it possible to perform successive dilutions. The liquid containing the solution to be diluted (liquid containing a reagent, and/or biological samples, and/or beads, and/or cells, etc.) is dispensed into the container R16. The objective of the protocol is to dilute the reagent (the sample, beads, cells, respectively). For this, the container R1 is filled with the dilution buffer (water, biological buffer, etc.). The chip is controlled by means such as means 40, 42 of
There may be for example 4 types of basic instructions:
Thus (
The new drop g3 thus formed can be stored, for example in container R3. The dilution operation is repeated to form a droplet g4 from R2 and a new drop g5 from R1, with the result being stored in container R4. This operation is repeated until concentrations C1, C1/7, C1/49, Ci/7n are obtained in each container R2 to Rn.
This situation is shown in
To summarise, the instructions to be provided to the control system 40, 42 of the fluidic component in order to perform 4 successive dilutions with storage of the liquids in the containers R2 to R16 are provided in the following table.
The process can be repeated for all of the 14 containers R2 to R15. It is also possible to form a plurality of drops with equivalent concentrations.
For example, it is possible to carry out 4 successive dilutions to obtain a concentration C1/2401, then repeat the dilutions but always from the same container R5. Thus, the other containers R7, R8, R9, and so on will be filled with a liquid volume with the same concentration C1/2401.
After coalescence, the drop can be moved on the bus 301 to homogenise and/or mix the liquids. Typically 12 to 20 movements on the electrodes of the bus are enough for an effective mixture. It is also possible to use the line selection electrodes to have the drops perform two-way movements between the containers and the bus 301 in order to agitate the liquids.
The same protocol can be carried out with cells. By implementing the invention, it is possible to manipulate drops containing only a few cells, or even a single cell. It is then possible to apply a biological protocol on this drop in order to study and/or analyse the behaviour of the cell. This protocol can be carried out in parallel on a very large number of drops. One of the applications is drug screening.
A microfluidic switching device 501, 502, 503 is used in combination with each relay.
Such a microfluidic switching device operates according to the following principles, which will first be explained in the context of an open configuration. Thus, we will consider the case, shown in
The reverse operation can then be performed, with the drop then returning to its initial position 2 and the conductor 12 is no longer at the potential of conductor 10.
In this operation, the drop 2 is stretched, but not moved. In addition, the contact is achieved by stretching the drop over the planar surface 8. A switching or a change in state therefore results from a stretching of the drop so as to put two lines 10, 12 in contact.
In the initial state, the drop 2 can be formed on a container electrode and be stretched over another neighbouring electrode 4-3.
From the logic perspective, it will be assumed that the potential 0 of the electrodes 4 causes the drop to be spread.
As seen in
Thus, a microfluidic logic switch is produced.
Another embodiment is shown in
A device according to the invention can also have a closed configuration, of the type shown in
In this case, shown in
In the embodiments already described, the drop is, by stretching or deformation, brought into contact with two conductors located parallel to the substrate 1 or located between the substrate 1 and the cap 11.
According to another embodiment of the invention (
A portion 107 and 107′, respectively, of layer 13 of the cap 11 is, for example, etched, so that a drop 2 of conductive liquid makes it possible to produce a contact with the conductor 11-2 and 11-2′, respectively (drop in stretched position 2′) of the cap. It is also possible to allow a very fine hydrophobic layer, for example on the order of several dozen nm for Teflon, to remain in area 107 and/or area 107′; it is then porous to electrical charges. It is then unnecessary, in this case, to completely etch the hydrophobic layer 13 in this area.
The thickness of the hydrophobic layer allowing a certain porosity for the charges, sufficient for circulation of the current with the counter electrode 11-2 and 11-2′, respectively, will depend on the material of the layer 13. In the case of Teflon, there are indications on this subject in the document of S.-K. Cho et al., “Splitting a liquid droplet for electrowetting-based microfluidics”, Proceedings of 2001 ASME International Mechanical Engineering Congress and Exposition, Nov. 11-16, New York. As regards Teflon, a layer of 20 nm, or for example less than 30 nm, is enough to allow charges to pass. For each hydrophobic and/or insulating material, a test can be conducted according to the thickness deposited in order to determine whether the desired potential is reached with regard to the electrode 15.
According to the invention, the switch from one state to another can be controlled by switching from a contact of the drop with an area of the layer 13 where the latter is inexistent or weak, to a contact of the drop with two areas of this layer where the latter is inexistent or weak.
According to yet another embodiment of the invention (
The two electrodes 4-2 and 4-4 are therefore used as contact areas for two states, one in which the drop 2 is only in contact with the electrode 4-2 and the other in which the drop 2 is in contact with the two electrodes 4-2 and 4-4. The switch from one to the other is performed by electrowetting by activation of electrodes located between the depassivated electrodes.
Finally, it is possible to combine the various embodiments above. For example, in
Thus, the switching can take place between the area 107 and the conductor 12.
Complex functions can be developed from one of the basic configurations disclosed above.
In this figure, at least 4 electrodes 4-1, 4-2, 4-1′, 4-2′ are concerned. The electrodes 4-1 and 4-1′ are respectively in state 1 and 0, while electrodes 4-2 and 4-2′ are initially at any potential Va.
Each of the two catenaries 10 and 10′ plays the same role, respectively for electrode 4-1 and for electrode 4-1′, as already explained above for the catenary 10 with respect to electrode 4-1.
Two states are thus possible.
When Va=1 (
When Va=0, (
The complement function explained above in relation to
This device can advantageously be used in a device according to the present invention.
In the diagram of
Each unit 501, 502, 503 is therefore a device making it possible to form a complement function of a voltage, called the input voltage. Such a device comprises two switching devices, each switching device comprising:
At least one of the two contact conductors of a switching device can comprise a depassivated electrowetting electrode 4-2, 4-4.
A switching device can also comprise a cap 11 with a hydrophobic surface 13 opposite the hydrophobic layer of the substrate, at least one of the two contact conductors comprising an electrode 11-2, 11-2′ arranged in the cap, a portion 107, 107′ of the hydrophobic surface of said cap either being etched or having a low enough thickness to allow electrical charges to pass.
The means for switching a drop can comprise means for switching a voltage applied to at least one electrowetting electrode, called a switching electrode, between a first value, for which the drop is not in contact with the second conductor and a second value, for which the drop is in contact with the second conductor.
A device for forming a complement function of a voltage (Va), called the input voltage, therefore comprises:
The conductive liquid used for the drops 2′, 21 used in a switching device can be a liquid, a conductive gel, or a material with a low melting point (for example: lead, tin, indium or silver or an alloy of at least two of these materials), which, by the phase change, causes a permanent or temporarily fixed contact (the phase change can indeed be reversible), or a conductive glue (hardening or solidifying by polymerisation, for example). The production of a permanent contact, or the blockage of a switch, can indeed be useful, so as not to provide an electrical supply the contactor or the logic functions while maintaining the spreading of the drop. Thus, the switch or the logic function consumes energy only during the change in state.
Number | Date | Country | Kind |
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04 51494 | Jul 2004 | FR | national |
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/FR2005/050570 | 7/11/2005 | WO | 00 | 12/28/2006 |