EP2931425B1 - Manipulation de la taille des gouttes liquides en microfluidique numérique - Google Patents

Manipulation de la taille des gouttes liquides en microfluidique numérique Download PDF

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Publication number
EP2931425B1
EP2931425B1 EP13770655.2A EP13770655A EP2931425B1 EP 2931425 B1 EP2931425 B1 EP 2931425B1 EP 13770655 A EP13770655 A EP 13770655A EP 2931425 B1 EP2931425 B1 EP 2931425B1
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EP
European Patent Office
Prior art keywords
electrode
container
liquid
electrodes
liquid droplet
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EP13770655.2A
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German (de)
English (en)
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EP2931425A1 (fr
Inventor
Marc N. Feiglin
Joseph Mamone
Michael Benjamin Franklin
Timothy Francis Lee
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Tecan Trading AG
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Tecan Trading AG
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Priority claimed from US13/784,168 external-priority patent/US8936708B2/en
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    • 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
    • B01L3/502715—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 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • 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
    • B01L3/502769—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 characterised by multiphase flow arrangements
    • B01L3/502784—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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B01L3/502792—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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics for moving individual droplets on a plate, e.g. by locally altering surface tension
    • 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
    • B01L3/5029—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures using swabs
    • 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
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0819—Microarrays; Biochips
    • 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
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • 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
    • B01L2400/0427—Electrowetting
    • 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
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • 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
    • B01L3/02—Burettes; Pipettes
    • B01L3/0241—Drop counters; Drop formers
    • B01L3/0268—Drop counters; Drop formers using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
    • 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
    • B01L3/52—Containers specially adapted for storing or dispensing a reagent
    • B01L3/527—Containers specially adapted for storing or dispensing a reagent for a plurality of reagents

Definitions

  • the present invention relates to manipulating the size of liquid droplets in digital microfluidics.
  • This technical field generally relates to the control and manipulation of liquids in a small volume, usually in the micro- or nanoscale format. Movement of small liquid volumes in a channel system is known per se as, e.g. being controlled by micro pumps in stationary devices or centripetal forces in rotating lab-ware.
  • a defined voltage is applied to electrodes of an electrode array, so that individual droplets are addressed by electrowetting.
  • electrowetting method please see Washizu, IEEE Transactions on Industry Applications, Volume 34, No. 4, 1998 , and Pollack et al., Lab chip, 2002, Volume 2, 96-101 .
  • a major advantage of the electrowetting technology is that only a small volume of liquid is required, e.g. a single droplet.
  • liquid processing can be carried out within considerably shorter time.
  • control of the liquid movement can be completely under electronic control resulting in automated processing of samples.
  • An apparatus with a single-sided electrode design for manipulating droplets is also known from US 6,911,132 B2 , all conductive elements being contained on a first surface on which droplets are manipulated.
  • An additional surface can be provided parallel with the first surface for the purpose of containing the droplets to be manipulated.
  • the apparatus enables droplet manipulating such as merging and mixing two droplets together, splitting a droplet into two or more droplets, sampling a continuous liquid flow by forming from the flow individually controlled droplets, and iterative binary or digital mixing of droplets to obtain a desired mixing ratio. Merging is performed using at least three aligned drive electrodes that initially are switched off, the first and third electrode each bearing a drop of liquid on top of the surface covering it.
  • the liquid droplet is positioned in the gap and moved within a non-polar filler fluid by consecutively applying a plurality of electric fields to a plurality of electrodes positioned on the opposite sites of the gap.
  • a drop meter with larger area defined by a hydrophobic surface of a control pad is connected via a small extension to a path or driveway of drive electrodes, the first one being a cutoff electrode and the second one being a control electrode.
  • a wetting potential is first applied to the cutoff electrode, the liquid which has covered the surface of the control pad thus spreading over the cutoff and drive electrodes. Consequently, the wetting potential is removed from the cutoff electrode, making it hydrophobic again. Part of the liquid moves back to the contact pad ad is replaced by the non-polar filler fluid. As a result, an isolated drop of liquid is separated and formed on the control electrode.
  • WO 2010/040227 A1 is known a hybrid digital and channel microfluidic device in the form of an integrated structure in which a liquid droplet may be transported by a digital microfluidics array and transferred to a microfluidic channel. Merging two liquid separate droplets by moving them on two separated driveway and then joining these electrode paths is also disclosed.
  • Figure 1 shows a schematic cross section and partial layout of an exemplified biological sample processing system 1 according to the first aspect of the present invention.
  • this system 1 comprises distinct single components which can be assembled to one unit, the system 1, in simple steps.
  • Such component is for example a container 2 which is comprised by the biological sample processing system 1.
  • the container 2 is accomplished for processing large volumes of liquid 18.
  • large volumes of liquid are understood to relate to liquid volumes up to 5 ml or up to 10 ml, depending on the sample to be hosted.
  • the large volume well is preferably designed to hold volumes up to 2 ml; if holding for example whole blood, the well preferably holds up to 5 ml.
  • the container 2 has a top side 3 and a base side 4. At its base side 4, the container 2 comprises protrusions 5. These protrusions 5 may be accomplished as parts of the container 2 extending downwardly at the base side 4. Alternatively these protrusions 5 may be attached to the base side 4 of the container 2 separately, for example by gluing, welding or other appropriate means to stably attach such protrusions 5 to the base side 4 of the container 2.
  • the container 2 comprises at least one well 6. This at least one well 6 is open at its top side 7.
  • a biological sample 9, a reaction reagent 10, or both can be positioned within this well 6.
  • the at least one well 6 has at least one opening 11. This opening is connected by a channel 12 of the container 2 with an orifice 13 of the container 2 at its base side 4.
  • a liquid 18 or a liquid droplet 19 is placed into the at least one well 6 (with or without a reaction reagent, and/or with or without at least parts of the biological sample 9), it can be transferred out of the well 6 through the channel 12.
  • the diameter of the channel 12 preferably is chosen such that the capillary force prevents liquid from leaking out the orifice 13 and that the liquids thus stay inside the at least one well 6 without need for a valve or any other closure of the channel 12.
  • the diameter of the channel 12 preferably is from 100 ⁇ m to 1 mm.
  • the biological sample processing system 1 comprises a flat polymer film 14.
  • This flat polymer film 14 could also be called a "plastic skin” as proposed by Yang et al. (2008) “Exchangeable, pre-loaded “Skin Depot” for digital microfluidics” at the MicroTAS meeting in San Diego, CA .
  • This flat polymer film 14 preferably has a lower surface 15 and a hydrophobic upper surface 16.
  • As a material for the thin polymer films food wraps, and stretchable wax films can be used.
  • the protrusions 5 keep the flat polymer film 6 at a distance "d" to the base side 4 of the container 2.
  • This distance "d” is set by the height of the protrusions 5 of the container 2, and defines at least one gap 17 when the container 2 is positioned on the flat polymer film 14.
  • the gap 17 between the upper hydrophobic surface 16 of the film 14 and the base side 4 of the container is sized to accommodate a liquid droplet.
  • this gap 17 is less than 2 mm.
  • the gap 17 is less than 1 mm.
  • the biological sample 9 preferably is contained a well 6. It can be mixed with a liquid 18, such as a buffer solution with or without lysis reagents.
  • the biological sample 9 may be displaced from the at least one well 6 (while kept within a liquid droplet 19), through the channel 12 of the container 2 onto the hydrophobic upper surface 16 of the flat polymer film 14. The liquid droplet 19 with the biological sample 9 is thus placed in the gap 17 between the film 14 and the container 2.
  • the displacement may be performed, without using valves, by a pressure applied, centrifugal force, or electrowetting against the capillary forces that prevent leaking of liquids out of the wells 6,6'.
  • other means may be used which are suitable to displace the liquid 18 or liquid droplet 19 from the well 6 onto the hydrophobic upper surface 16 of the flat film 14.
  • These means for displacement may also be used for transferring a reaction reagent 10, which is stored in a well 6' of the container 2, onto the upper surface of the film. When displacing liquids from the wells 6,6' onto the upper polymer film surface 16, excess air form the gap may be vented e.g. via an empty well 6'.
  • the biological sample processing system 1 comprises furthermore a liquid droplet manipulation instrument 20.
  • This instrument 20 comprises at least one electrode array 21, a substrate 22 which supports the at least one electrode array 21 and a control unit 23.
  • the liquid droplet manipulation instrument 20 is accomplished so that the container 2 and the film 14 can be reversibly attached to the instrument 20. Thereby, the lower surface 15 of the film 14 abuts the electrode array 21.
  • the biological sample processing system 1 enables the displacement of a liquid droplet 19 from the at least one well 6 of the container 2 onto the upper surface 16 of the flat polymer film 14 and accordingly above the at least one electrode array 21.
  • the electrode array 21 is accomplished to induce movements of the liquid droplets 19.
  • the instrument 20 is accomplished to control a guided movement of said liquid droplet 19 on the upper surface 16 of the flat polymer film 14 by electrowetting and to process there the biological sample 9.
  • Typical biological samples 9 which are processable by a biological sample processing system 1 are nucleic acids or proteins.
  • nucleic acids are used for processing.
  • Such nucleic acids comprise DNA (desoxyribonucleic acid, for example genomic DNA, cDNA, mtDNA), RNA (ribonucleic acid, for example mRNA), being single- or double stranded, and derivatives thereof (for example artificially labeled nucleic acids.
  • These biological samples 9 may be contained in tissue samples such as oral mucosa cells or hair follicles.
  • the biological samples 9 may be contained in a liquid, such as samples of body fluids such as blood, urine, sputum etc.
  • a biological sample 9 of interest can be processed by a biological sample processing system 1 according to the present invention independent of its origin.
  • the selection of required reaction reagents 10 should be adopted based on the material which comprises the biological sample 9. It is also possible to load an already purified biological sample 9 into the at least one well 6 of the container 2. In this case, a purification step is not necessarily required during the processing within the biological sample processing system 1.
  • a container 2 is shown to comprise one well 6 sized to accommodate a swab head, and further wells 6' which are different from the sample well 6 in size.
  • the container 2 comprises at least one sample well 6 and six smaller wells 6' for storing reaction reagents. These wells 6' are preferably sized to store reaction reagents 10 and other required liquid such as buffers.
  • a typical size of such a well 6' for storing reaction reagents 10 has a diameter of about 5 mm and a height of about 40 mm.
  • the size of each well 6,6' of the container 2 may be adopted individually according to the requirements given by the underlying question to be solved.
  • the container 2 of the biological sample processing system 1 comprises at least one well 6 which is accomplished as storage for a reaction reagent 10.
  • This embodiment is particularly preferred in case the container 2 comprises already one well 6 for positioning a biological sample 9.
  • the container comprises at least two wells 6, one well 6 for positioning the biological sample 9 and one well 6' for storing a reaction reagent.
  • Stored reagents 10 comprise reagents selected from a group comprising reagents for performing cell lysis, reagents for performing nucleic acid purification, reagents for performing nucleic amplification and reagents for performing sequencing of nucleic acids.
  • the cellular integrity is disrupted by opening cell membranes.
  • This step can be performed using for example enzymatic activity or chemical lysis.
  • other procedures to disrupt cellular integrity may be suitable.
  • the thermostable protease EA1 manufactured by ZyGemTM Corporation (Waikato Innovation Park, Ruakura Road, Hamilton, New Zealand) should be mentioned at this place as a suitable enzyme for performing cell lysis.
  • cell lysis may be carried out using Proteinase K, or chemical lysis, both procedures also well known in the art.
  • the buffer matching to the used enzyme can be chosen by a skilled person without the need of special efforts and is regarded to be based on the general knowledge in the art, too. As the procedure of performing cell lysis is well known to a skilled person, it should not be described here in more detail.
  • Sequencing of specifically amplified DNA is a well known tool to further characterize the selected DNA.
  • Major sequencing principles are known in the art, sequencing by amplification and sequencing by hybridization. Sequencing by amplification involves a PCR-related process using however labeled stop-primer which terminate the extension process randomly. The resulting end-labeled fragments are then used for determining the sequence of the template.
  • Sequencing by hybridization involves the linkage of labeled primer to a matrix. Primer are selected so that the overlap partially in their sequence. After hybridization of a target DNA to said primer, sequence can be determined by the analysis of primer sequenced to which the target is bound.
  • the labeled primers are preferably linked to the hydrophobic upper surface 16 of the flat polymer film 14 prior to the start of sample processing. Most preferably, the labeled primers are linked prior to the release of the system into the trade.
  • the number of wells 6,6' is dependent on the type of reaction system used (required reagents, processing steps) and the number of analysis required (number of sequences/loci to be analyzed, i.e. STR, SNP, mtDNA) and may be adopted by a skilled person based on general knowledge in the art.
  • the primer for the amplification should be stored in the container 2, preferably the container 2 comprises one primer well per loci to be analyzed for the amplification process.
  • the container preferably comprises 16 primer wells 6' for the amplification.
  • the primer required for the amplification step may be available in dried form on the hydrophobic upper surface 16 of the flat polymer film 14, so that for storing primer, no separate well 6' would be required here in the container 2.
  • the primer may in this case be re-suspended on the film 14 using a buffer held in a well 6'.
  • the number of wells 6' required storing reaction reagents 10 and buffer may similarly be adopted.
  • a container 2 according to the present invention is shown in a top view, having one well 6 for positioning the biological sample 9 and six further wells 6' for storing reaction reagents 10.
  • the biological sample 9 When cell lysis is performed directly within the well 6 for positioning the biological sample 9, the biological sample 9 is set free from the cellular context, and preferably released into a liquid 18, which therefore is a reaction solution resulting from the cell lysis.
  • the liquid can be chosen according to the following procedural steps, and added into the well via the top side 7 of the well 6.
  • the biological sample 9 should be contained at least in parts in a liquid for the further processing using the biological sample processing system 1 according to the first aspect of the present invention.
  • the liquid 18 or a liquid droplet containing at least parts of the biological sample 9 is then displaced for further processing from the well 6 through the channel 12 of the container 2 onto the hydrophobic upper surface 16 of the flat polymer film 14.
  • the displacement preferably performed, without using valves or other moveable means, by a pressure applied, by centrifugal force, or by electrowetting. All these preferred displacement means act against the capillary forces that prevent leaking of liquids out of the wells 6,6'. However, also other means may be used which are suitable to displace the liquid 18 or liquid droplet 19 from the well 6 onto the hydrophobic upper surface 16 of the flat film 14.
  • the container 2 and the flat polymer film 14 are reversibly attached to the liquid droplet manipulation instrument 20, with the lower surface 15 of the film 14 abutting the electrode array 21. Accordingly, the liquid droplet 19 is displaced from the well 6 above the electrode array 21.
  • the liquid droplet 19 may be moved in a guided manner by the liquid manipulation instrument 20 by electrowetting. The movement is controlled to achieve the selected processing of the biological sample 9 contained within said liquid droplet 19 and to carry out this processing at preferred sites of the electrode array.
  • the liquid droplet 19 is moved in the gap 17 within an immiscible system liquid 32.
  • This variant is the preferred embodiment, when performing PCR on the biological sample 9 contained in the at least one liquid droplet 19.
  • PCR requires the exposure of such a liquid droplet 19 to different temperatures, including the denaturation step at about 98°C, evaporation of liquid may be prevented or at least considerably reduced with the use of such an immiscible system liquid 32.
  • Preferred system liquids immiscible with the liquid droplet 19 are selected e.g. from silicon oil, hexadecane and benzene.
  • Positioning elements 25 accomplished as at least one groove of the container 2 and at least one elevation extending from the instrument 20 are presented in Figure 1 (with the groove at the base side 4 of the container 2), Figure 2A (with two triangular-shaped grooves in the lateral area of the container) and in Figure 2B (with two semi-circular shaped grooves in the lateral area of the container 2).
  • a Peltier element for heating the well 6 for positioning the biological sample 9 such Peltier element can be accomplished as an elevation extending from the instrument 20, its position may be chosen so that for example, is specifically provides the well 6 (whether this is central or not) with a defined temperature.
  • other means for positioning the container 2 on the liquid droplet manipulation instrument 20 in a defined configuration may as well be used which are well known to a skilled person, and should not be described in more detail here.
  • a liquid droplet 19 on the flat polymer film 14 may either contact only the hydrophobic upper surface 16 of the film 14 or may contact both, the hydrophobic upper surface 16 of the film 14 and the base side 4 of the container 2.
  • the contact surfaces of such a liquid droplet 19 may be influenced by the sizing of the gap 17 (thus, sizing the protrusions 5) or by sizing the liquid droplet 19.
  • the container 2 is preferably made by injection molding. In this way, the production costs may be reduced despite the achievable high manufacturing quality and the container 2 can be utilized as a low cost disposable.
  • Such a single-serving container 2 is suited to be sold for various applications and can be equipped with a specific set of reaction reagents 10.
  • the container 2 is preferably made either of an electrically insulating material 26, of an electrically conductive material 27, or of a combination of both an electrically conductive and an electrically insulating material 26,27. When made of two different materials, a two step injection process is preferred.
  • the core of the container 2 is made of an electrically insulating material 26, wherein the regions surrounding the wells 6,6' are made from an electrically conductive material 27.
  • the surrounding regions made of the conductive material 27 are separated from each other by the insulating material 26.
  • These surrounding regions made of conductive material 27 may form at the base side 4 of the container 2 a nozzle 47, which slightly extends into the gap 17 (see Figure 1 ).
  • the advantage provided by such a nozzle is the possibility to distinctly produce and deliver a liquid droplet 19 into the gap without the droplet 19 contacting the surface of the base side 4 of the container 2. Furthermore such a nozzle may enable a directed delivery of the droplet 19 into the gap 17.
  • each electrically conductive region 27 of the container 2 may be individually contacted electrically. This allows the conductive regions to be addressed by a voltage control 29 and provided with an individual voltage.
  • a voltage control 29 may be provided with an individual voltage.
  • one or more liquid drops may be displaced to the hydrophobic upper surface 16 of the flat polymer film 14 using the principle of electrowetting.
  • the displacement can be done for each well individually, so that reaction reagents 10 or liquids containing the biological sample 9 may be individually displaced at the time they are required on the film 14.
  • the at least one well 6 for positioning a biological sample 9 is arranged towards the outer, lateral side 28 of the container 2.
  • the at least one well 6 is additionally surrounded from an electrically conductive material 27.
  • the electrically conductive surrounding 27 is in this variant extends to form the major part of the core of the container 2.
  • major parts of the base side 4 of the container 2 are made of electrically conductive material 27, too.
  • This allows processing a liquid droplet 19, which is positioned on the hydrophobic upper surface 16 of the film 14 and which contacts the base side 4 of the container 2, by electrowetting using the electrically conductive parts of the base side 4 of the container 2 as a ground electrode. Accordingly, in this variant, the guided movement of the liquid droplet 19 may be further stabilized.
  • part of the container 2 is made from a thermally isolating material or that thermally insulating gaps are provided around the zone of higher temperature.
  • the container 2 comprises means for identification 30, which are selected from a group comprising a barcode and an RFID (radiofrequency identification) tag or another integrated chip.
  • means for identification 30 are well known to the person skilled in the art, they should not be described in more detail here.
  • Identification means 30 are especially preferred when the container 2 of the biological sample processing system 1 is used in an automated manner, while storing information for example about the biological sample positioned in a well 6 of the container 2. In addition, tracking of a particular sample is possible even in a large laboratory system.
  • this well 6 preferably comprises retention means 31 for preventing the solid substrate 24 to block the opening 8 of said well 6.
  • the retention means 31 are selected from a group comprising a filter, a frit (see Fig. 1 ) and relief structures (see Fig. 2B ). However, other retention means 31 well known in the art may be used for these purposes.
  • the analyzing area 33 is positioned above that region of the hydrophobic upper surface 16, which is accomplished to the processing of a biological sample 9 using sequencing, especially preferred when as the sequencing method sequencing by hybridization method is performed.
  • optical devices 38 are selected from a group comprising a standard microscope, a camera system, a light guiding system such as fiber optics, a scanner, and adaptations or combinations thereof.
  • a camera, a simple CCD or a PMT (Photo Multiplier Tube) is used together with a light source, such as an LED, which serves as an excitation source for fluorescent tags on the film 14.
  • a light guiding system is used, the excitation- and/or measurement device may be located aside of the container 2.
  • automatic sample processing and final analysis can be carried out on the same polymer film 16 and on the electrode array 21.
  • the container 2 preferably comprises a support rim 45 when having an analyzing area 33.
  • This support rim 45 extends along the outer lateral side 28 of the container 2 while bordering the analyzing area 33. Accordingly, this support rim 45 may also comprise one or more protrusions 5 at its bottom side, which are attached to the hydrophobic upper surface 16 of the flat polymer film 14.
  • This support rim 45 supports the container 2 having a cut-out when positioned on the film 14.
  • a multitude of containers 2 having at least one analyzing area 33 is arranged in such a way, that each analyzing area 33 is easily accessible by one optical device 38.
  • One possible way would be an essentially circular arrangement of the containers 2 around a rotary optical device 38.
  • the containers 2 can be stored in vertical or horizontal rows of adequate carrier, and the optical device 38 or the carrier with the row of container 2 are shifted manually or automatically into a position in which the analyzing 33 is accessible by the optical device 33.
  • Both, the container 2 and the flat polymer film 14 can be provided to the user either as separate components that remain to be assembled when the processing of a biological sample 9 is to be started.
  • these two components can be provided as a cartridge 40.
  • the cartridge comprises both, container 2 and the flat polymer film 14, which are attached to one another for example by gluing or welding or other appropriate means to stably attach these two components.
  • the container 2 or the cartridge 40 comprises a cover 43 for protecting the wells 6,6' and their content from outside influences.
  • a cover 43 may be sealingly attached to the top side 7 of the container 2.
  • the attachment may be reversible.
  • the cover 43 is a thin film, which optionally is made of a pierceable material. In this way, the wells 6,6' of the container 2 may be pre-loaded. Safe storage is allowed by applying the film cover 43 onto the container 2. Only upon start of the sample processing, the film cover 43 is pierced open and the wells 6,6' of the container 2 are accessible for the user. Additionally, the container 2 or the cartridge 40 can be covered with a cover 43 as well.
  • the present invention relates to a liquid droplet manipulation instrument 20.
  • this liquid droplet manipulation instrument 20 is accomplished to be used in the biological sample processing system 1 according to the first aspect of the present invention.
  • the liquid droplet manipulation instrument 20 may be used independently of the biological sample processing system 1.
  • the control unit 23 preferably comprises a power supply 44.
  • This power supply 44 provides at least the central processing unit 36 and the voltage control 29 with electric power.
  • the power supply 44 may additionally provide also other elements with electric power.
  • the control unit 23 is capable to define a path for a guided movement of a liquid droplet 19 by the selection of a series of subsequent drive electrodes 35'. Thereby at least one of these selected drive electrodes 35' is subsequently provided with a drive voltage pulse along said path, under the control of the control unit 23. Furthermore the control unit 23 is accomplished to essentially simultaneously provide at least one electrode 35", which is adjacent to the pulsed drive electrode 35' and different to the selected drive electrode 35' of the path, with a ground voltage pulse.
  • Such a path for a guided movement of the liquid droplet is each shown in the Figures 3A to 3D .
  • the subsequent selected drive electrodes 35' are indicated.
  • the actual drive electrode 35' is shown to be that electrode 35, upon which the liquid droplet 19 is positioned.
  • the direction of the planned guided movement of the liquid droplet 19 is indicated with an arrow. In that direction, subsequent electrodes 35' along the path will be provided with a drive voltage pulse.
  • the size, respectively the diameter of the liquid droplet 19 slightly exceeds the diameter of an electrode 35.
  • the liquid droplet touches not only the actual drive electrode 35' but slightly touches simultaneously the subsequent electrode 35' which will become the next actual drive electrode 35'.
  • the adjustment of electrode size in relation to liquid droplet sizes is within the knowledge of the person skilled in the art and should not be repeated here.
  • the actual size and design of the electrodes and the desired size of the liquid droplets 19 must be in accordance with each other and with the praxis of electrowetting.
  • the presence of at least one ground electrode 35" adjacent to the liquid droplet 19 to be moved provides a stabilizing effect to its movement.
  • the Figures 3A-3D indicate those electrodes 35" that might be provided with a ground voltage pulse. Those ground electrodes 35" are preferably adjacent to the pulsed drive electrodes 35' and different or identical to the selected drive electrodes 35' of the path.
  • the provision with the ground voltage pulse is preferably carried out essentially simultaneously to the provision with the drive voltage pulse. Alternatively, the provision with the ground voltage pulse is to be carried out simultaneously to the provision with the drive voltage pulse.
  • control unit 20 is accomplished to provide at least two electrodes 35" which are adjacent to the pulsed drive electrode 35' and different to the selected drive electrode 35' of the path with a ground voltage pulse.
  • these at least two selected ground electrodes 35" are subsequent electrodes 35 on the same side of the path.
  • ground electrodes 35" may be selected from electrodes 35 along the path, adjacent to the path and adjacent to the liquid droplet 19.
  • the selected ground electrodes 35" are on the same side of the path.
  • at least two first electrodes 35" are preferably selected from one side of the path.
  • the remaining electrodes 35" of that group may however be selected from that side of the path being opposite to the first two ground electrodes 35" of that group.
  • a group of ground electrodes 35" are selected from two sides of the path, they are provided with the ground voltage pulse essentially simultaneously or simultaneously to the pulsed drive electrode 35'.
  • the other electrodes 35" may be adjacent to the path and ahead of the liquid droplet 19, adjacent to the path and behind the liquid droplet 19 or both.
  • a group of 2 or more electrodes 35 may be provided with a drive voltage pulse essentially simultaneously. In this case, a liquid droplet 19' of a larger volume may be moved. However in this variant it is preferred that essentially simultaneously or simultaneously a group of 2 or more electrodes 35 are provided with a ground voltage pulse to sufficiently support the liquid droplet 19' with the larger volume.
  • control unit is accomplished to provide at least one selected electrode with a stop voltage pulse for generating a stop electrode 35"'.
  • the provided stop voltage pulse is different to the drive voltage pulse and the ground voltage pulse.
  • the voltage pulses for defining a selected electrode 35 as a drive electrode preferably are between 20 and 100 V.
  • the voltage pulses for defining a selected electrode 35 as a stop electrode preferably are between -50 V and +50 V.
  • selected stop electrodes 35'" are adjacent to the path, different to the selected drive electrode 35' of the path and different to the at least one selected ground electrode 35" adjacent to the path.
  • stop electrodes 35'" are selected from such electrodes 35 adjacent to the path, where the path provides a change of direction for the liquid droplet 19 movement.
  • a stop electrode 35''' supports the direction change of the liquid droplet movement along the path.
  • FIGS 3A-3D show exemplarily possible positions of stop electrodes 35''' along the path.
  • at least one electrode 35'" along the path at a place of direction change is provided with a stop voltage pulse.
  • more than one electrode 35'" in that area may be selected to be provided with a stop voltage pulse, as shown in Figure 3D .
  • two ore more electrodes are selected as stop electrode 35''' at the point of direction change to support the liquid droplet movement.
  • FIGS 3B and 3C show exemplary a virtual grid of the electrode array 21.
  • Each grid point 39 of the virtual grid is established by the geometrical centre of each electrode 35 of the electrode array 21.
  • Figure 3B shows a hexagonal grid according to the hexagonal shape and dense packing of each electrode 35 of the electrode array 21.
  • Figure 3C shows an orthogonal grid based on the orthogonal arrangement of electrode 35, this time exhibiting essentially circular shape.
  • subsequent electrodes for defining the path, subsequent selected ground electrodes 35", and/or subsequent selected stop electrodes 35''' are defined by the closest distance between two grid points 39 of that virtual grid. In this way a continuous liquid droplet movement may be ensured.
  • the hexagonal arrangement of the electrodes array 21 is preferred over the orthogonal arrangement because of the higher degree of freedom.
  • Figure 1 shows exemplarily the position of the electrodes 35 in relation to the substrate 22.
  • the electrodes 35 of the electrode array 21 are positioned in relation to the substrate 22, so that the upper surface of the electrodes 35 are aligned substantially flush with the upper surface of the substrate 22.
  • the electrodes 35 of the electrode array are positioned within the substrate 22 and enclosed by it (see left hand side on Fig. 1 ). It is preferred to position the electrodes 35 as close to the liquid droplets 19 as possible in order to be able to reduce the voltage necessary for electrowetting.
  • electrodes 35 flush with the upper surface of the substrate 22 and very thin polymer films are particularly preferred.
  • a material for the thin polymer films e.g. food wraps, and stretchable wax films can be used.
  • the liquid droplet manipulation instrument 20 is accomplished to accommodate a container 2 for large volume processing and to simultaneously accommodate a flat polymer film 14 with a hydrophobic upper surface 16.
  • a system is formed suited for biological sample processing of a sample 9 positioned within the container 2.
  • Such a system preferably corresponds to the biological sample processing system 1 according to the first aspect of the present invention.
  • the liquid droplet manipulation instrument 20 is accomplished to accommodate a cartridge 40.
  • Said cartridge comprises a container 2 and a flat polymer film 14 as previously described herein.
  • Container 2 and film 14 of the cartridge 40 are attached to one another by gluing or welding, or by other appropriate means to stably connect the container 2 and the film 14.
  • a biological sample processing system 1 according to the first aspect of the present invention may be formed.
  • the liquid droplet manipulation instrument 20 comprises at least two ore more electrode arrays 21.
  • the electrode arrays 21 are arranged essentially horizontal within the liquid droplet manipulation system 20.
  • the instrument 20 is accomplished to accommodate at least two or more containers 2 together with two or more flat polymer films 14, or to accommodate at least two or more cartridges 40.
  • the container 2 and the film 14 or the cartridge 40 may be positioned essentially above the electrode array.
  • the biological sample processing system 1 comprises a liquid droplet manipulation instrument 20 according to the second aspect of the present invention and as discussed in detail above.
  • the embodiment of the liquid droplet manipulation instrument 20 as well as the embodiment of the biological sample processing system 1 may be chosen by selecting the various features discussed above, depending on the question addressed. If not stated otherwise, the various features presented within this application may all be combined with each other.
  • the positioning elements 25 of the liquid droplet manipulation instrument 20 are comprised by the reception element 46.
  • Figure 3A shows an electrode array 21 according to such an especially preferred embodiment of a biological sample processing system 1.
  • This Figure shows an enlarged top view of a distinct section of Figure 1 , indicated as a rectangle with a dotted line.
  • the position of the at least one well 6 for positioning a biological sample 9 in relation to the defined path on the electrode array 21 is indicated by a dotted circular line in Figure 3A .
  • the opening 11 at the bottom of the at least one well 6, the passage of the channel 12 or the orifice 13 at the base side 4 of the container 2 respectively are indicated as an inner circle of a dotted line.
  • a liquid 18 or a liquid droplet 19 is transferred from the well 6 through the channel 12 on the hydrophobic upper surface 16 of the flat polymer film 14, namely above the electrode array 21 of the liquid droplet manipulation instrument 20.
  • a liquid portion 19' is indicated in the center of the electrode array 21 shown. This liquid portion 19' covers at least one selected drive electrode 35' from the electrode path. Preferably, the liquid portion covers at least partially subsequent electrodes 35' from the path. According to Figure 3A , the liquid portion 19' covers additionally electrodes 35" selected to be provided with a ground voltage pulse.
  • a liquid droplet 19 is separated from the liquid portion 19' by the provision of a drive voltage pulse to an electrode 35' subsequent to the initial drive electrode 35' along the path.
  • the liquid droplet 19 is then guided on along the path in a first direction, and after a direction change, in a second direction. At the position of the direction change, a stop electrode 35''' is generated to stabilize the direction change.
  • the liquid droplet manipulation instrument 20 of the biological sample processing system 1 comprises preferably at least one magnet 41.
  • This magnet 41 controls the magnetic beads during the processing in the gap 17 on the upper hydrophobic side 16 of the flat polymer film 14.
  • Suitable magnets may be electromagnets or permanent magnets.
  • the magnet 41 is arranged preferably on that side of the substrate 22 of the instrument, which is not covered by an electrode array 21.
  • the magnet 41 is alternatively arranged preferably on that side of the substrate 22 of the instrument 20, which is not abutted by the flat polymer film 14.
  • the liquid droplet manipulation instrument 20 of the biological sample processing system 1 preferably comprises at least one heating element 42.
  • This heating element 42 is preferably arranged on that side of the substrate 22 of the instrument 20, which is opposite to the side of the substrate 22 being abutted with the flat polymer film 14.
  • the at least one heating element 42 is accomplished to provide at least one temperature zone with a predefined temperature on the upper hydrophobic surface 16 of the flat polymer film 14. If the liquid droplet manipulation device 20 comprises one heating element 42, PCR may be performed by keeping the liquid droplet 19 comprising a biological sample 9 within the single temperature zone, while changing the temperature within that single zone accordingly.
  • a PCR may be done by moving the liquid droplet 19 comprising a biological sample 9 between the two zones, wherein the temperature of each zone is adopted according to the temperature required for the cycling steps.
  • the biological sample processing system 1 comprises in an especially preferred variant at least three heating elements 42 for providing at least three different temperature zones on the upper hydrophobic surface 16 of the flat polymer film 14.
  • Figure 1 shows a biological sample processing system 1 having three heating elements 42 underneath the support substrate 22 and opposite to the side being abutted with the flat polymer film 14.
  • Each temperature zone has a predefined temperature to enable a PCR being performed on the upper hydrophobic surface 16 of the flat polymer film 14.
  • one temperature zone comprises a temperature for denaturizing double stranded nucleic acid
  • one temperature zone comprises a temperature enabling the annealing of pre-selected primer
  • one temperature zone comprises a temperature enabling a polymerase to elongate the annealed primer to the full strand.
  • the biological sample processing system 1 may comprise a fourth heating element 42 providing a temperature of about 4°C. Utilizing at least three heating elements 42 underneath the support substrate 22 has the advantage that the selected temperatures can be kept constantly over the entire reaction time and the droplets can be moved from one temperature region to another. This movement allows for rapid temperature changes within the droplets 19, which are much faster than achievable by changing the temperature of the heater element 42 while keeping the droplet 19 in place.
  • a layer of low vapor pressure liquid connects the lower surface 15 of the flat polymer film 14 with the upper surface of the at least one electrode array 21 to reduce the formation of air bubbles in-between.
  • the low vapor pressure liquid is silicon oil; however, other low vapor pressure liquids may be used as well.
  • FIG. 4 shows top views of a grid-like electrode array 21 according to a first preferred embodiment and similar to the array of Fig. 3A with rectangular electrodes 35.
  • a particularly preferred liquid droplet manipulation instrument 20 is shown, the liquid droplet manipulation instrument 20 comprising:
  • control unit 23 of the liquid droplet manipulation instrument 20 the control unit 23 of the liquid droplet manipulation instrument 20:
  • Fig. 4A shows a liquid portion 19' of a larger volume that covers about 18 pulsed drive electrodes 35' of the same electrode array 21.
  • these 18 electrodes 35 have been activated for collecting this liquid portion 19' of a larger volume, the activated electrodes 35' attracting the liquid or at least keeping it in a stable volume.
  • this liquid portion 19' of a larger volume is kept on the hydrophobic upper surface 16 of a flat polymer film 14 and within a gap 17, the flat polymer film 14 being exposed to the substrate 22 with the at least one electrode array 21 (compare with Fig. 1 ).
  • more electrodes 35 are defined as ground electrodes 35" or stop electrodes 35'" (shown in white). All electrodes 35 are operatively connected to the electrode selector 34 of the control unit 23 (only partially shown in Fig. 4A and present but not shown in the Figs. 4B and 4D ).
  • Fig. 4B shows a group of two electrodes 35" that are identical with previously pulsed drive electrodes 35'that now are provided with a ground voltage pulse. This causes the liquid portion 19' of a larger volume to draw back to the activated electrodes 35' and induces a partial separation (see double arrow) of the liquid portion 19' of a larger volume into two smaller portions 19' that still cover at least two pulsed drive electrodes 35' of the same electrode array 21.
  • Fig. 4C shows a group of three electrodes 35" that are identical with previously pulsed drive electrodes 35' and that now are provided with a ground or stop voltage pulse (the latter even enhancing the separation effect). This causes the liquid portion 19' of a larger volume to now completely separate (see one-sided arrows to the left and right) into two smaller portions 19' that still cover at least two pulsed drive electrodes 35' of the same electrode array 21.
  • separation of a small volume i.e. a single liquid droplet 19 can also be performed when using the same particularly preferred liquid droplet manipulation instrument 20 (see Fig. 5 ).
  • Figure 5 shows top views of a grid-like electrode array according to a second preferred embodiment and similar to the array of Fig. 3A with rectangular electrodes.
  • a liquid portion 19' of a larger volume covers about 6 pulsed drive electrodes 35' of the same electrode array 21.
  • an additional drive electrode 35' of the same electrode array 21 is activated so that the liquid portion 19' now covers about 7 drive electrodes 35' (see Fig. 5B , on-sided arrow to the right). Then, opposite to the direction, where the additional drive electrode 35' was activated in Fig. 5B , another drive electrode 35' is activated and a group of two electrodes 35" identical with previously pulsed drive electrodes 35' are provided with a ground or stop voltage pulse. This causes a liquid droplet 19 to be dispensed and the remaining liquid portion 19' of a larger volume to move in different directions (see double arrow in Fig. 5C ). Providing the group of two electrodes 35" with a stop pulse even enhances the desired separation effect.
  • a combination of using a drive path of three rows of electrodes 35 (as e.g. shown in Fig. 4 ) and an electrode path of a single electrode row that closely reaches to the electrode array 21 (compare with Fig. 3A ) is preferably used for dispensing small liquid volumes, i.e. single liquid droplets 19 when utilizing the same particularly preferred liquid droplet manipulation instrument 20 (see Fig. 6 ).
  • Figure 6 shows top views of a grid-like electrode array 21 according to a third preferred embodiment and similar to the array of Fig. 3A with rectangular electrodes 35 and an electrode path adjacent to the electrode array 21.
  • All electrodes 35 are operatively connected to the electrode selector 34 of the control unit 23 (only partially shown in Fig. 6A and present but not shown in the Figs. 6B to 6F ).

Claims (12)

  1. Procédé pour séparer une gouttelette de liquide (19) d'une partie de liquide (19') dans un système de traitement d'échantillon biologique (1) comprenant un instrument de manipulation de gouttelette de liquide (20) comprenant :
    • au moins un réseau d'électrodes (21) avec des électrodes (35) pour induire par électromouillage un mouvement d'au moins une gouttelette de liquide (19) qui couvre une seule électrode (35) et d'une partie de liquide (19') d'un volume plus important qui couvre plusieurs électrodes (35) du réseau d'électrodes (21) ;
    • un substrat (22) supportant le au moins un réseau d'électrodes (21);
    • une unité de commande (23) capable de définir une trajectoire pour un mouvement guidé des gouttelettes de liquide (19) et des parties de liquide (19') ;
    • au moins une commande de tension (29) ;
    • au moins un sélecteur d'électrode (34) raccordé à la au moins une commande de tension (29) pour sélectionner individuellement chaque électrode (35) du au moins un réseau d'électrodes (21) et pour fournir à l'électrode sélectionnée (35) une impulsion de tension individuelle qui est sélectionnée à partir d'un groupe comprenant une tension de commande, une tension de terre et une tension d'arrêt ;
    • une unité de traitement centrale (36) pour commander le sélecteur d'électrode (34) et la commande de tension (29) ;
    • un récipient (2) ayant un côté supérieur (3) et une base (4) avec des saillies (5) réparties sur la base (4) ; et
    • un film polymère plat (14) ayant une surface inférieure (15) et une surface supérieure hydrophobe (16) qui est maintenue à une certaine distance (d) de la base (4) du récipient (2) par les saillies (5), la distance (d) définissant au moins un espace (17) lorsque le récipient (2) est positionné sur le film polymère plat (14), en butée contre ce dernier avec ses saillies (5) ;
    dans lequel le procédé comprend les étapes consistant à :
    (a) fixer de façon réversible le récipient (2) et le film polymère plat (14) à l'instrument de manipulation de gouttelette de liquide (20) de façon à ce que la surface inférieure (15) du film polymère plat (14) vienne en appui contre le au moins un réseau d'électrodes (21) ;
    (b) placer dans l'espace (17) une partie de liquide (19') sur la surface supérieure hydrophobe (16) du film polymère plat (14) et au-dessus du au moins un réseau d'électrodes (21) ;
    (c) fournir une impulsion de tension de commande à au moins une première électrode de commande (35') le long d'une trajectoire d'électrode définie, déplaçant ainsi la partie de liquide (19') d'un volume plus important dans une première direction ;
    (d) fournir à au moins une des premières électrodes de commande (35') une impulsion de tension de terre ou d'arrêt ; et
    (e) fournir une impulsion de tension de commande à un groupe de deux électrodes de commande ou plus (35') du réseau d'électrodes (21) afin de déplacer ladite partie de liquide (19') d'un volume plus important dans une deuxième direction opposée à la première direction.
  2. Procédé selon la revendication 1, dans lequel à l'étape (d), deux des premières électrodes de commande (35') reçoivent une impulsion de tension de terre ou d'arrêt pour séparer une gouttelette de liquide (19) de ladite partie de liquide (19') d'un volume plus important.
  3. Procédé selon la revendication 2, dans lequel à l'étape (d), deux des premières électrodes de commande (35') reçoivent une impulsion de tension d'arrêt pour séparer une gouttelette de liquide (19) de ladite partie de liquide (19') d'un volume plus important.
  4. Procédé selon la revendication 1, dans lequel le récipient (2) et le film polymère plat (14) sont conçus sous la forme d'une cartouche (40) qui comprend les deux, le récipient (2) et le film polymère plat (14), qui sont fixés l'un à l'autre par collage ou soudage.
  5. Procédé selon la revendication 1, dans lequel le récipient (2) et le film polymère plat (14) sont conçus sous la forme d'éléments séparés qui doivent être assemblés lorsque le traitement d'un échantillon biologique (9) démarre.
  6. Procédé selon la revendication 1, dans lequel le film polymère plat (14) est un film polymère plat électriquement isolant.
  7. Procédé selon la revendication 1, dans lequel il est prévu une couche diélectrique sur une surface du réseau d'électrodes (21).
  8. Procédé selon la revendication 5, dans lequel la couche diélectrique fait partie du substrat (22) de l'instrument de manipulation de gouttelette de liquide (20) ou fait partie du film polymère plat (14).
  9. Procédé selon la revendication 1, dans lequel la gouttelette de liquide (19) ou la partie de liquide (19') sont déplacées dans l'espace (17) dans un liquide de système non miscible (32).
  10. Procédé selon la revendication 1, dans lequel le récipient (2) est un récipient (2) pour le traitement de volumes importants et comprend au moins un puits (6) ouvert sur le dessus (3) pour venir y positionner un échantillon biologique (9) et/ou un réactif (10), et dans lequel le au moins un puits (6) comprend un fond (8) avec au moins une ouverture (11), le récipient (2) comprenant en outre un canal (12) raccordant l'ouverture du puits (11) à un orifice (13) sur la base (4) du récipient (2).
  11. Procédé selon la revendication 10, dans lequel au moins une gouttelette de liquide (19) provenant du au moins un puits (6) est déplacée à travers le canal (13) du récipient (2) sur la surface supérieure hydrophobe (16) du film polymère plat (14) et au-dessus du au moins un réseau d'électrodes (21), et dans lequel la gouttelette de liquide (19) est traitée par un mouvement guidé sur la surface supérieure hydrophobe (16) du film polymère plat (14) par électromouillage commandé par l'instrument de manipulation de gouttelette de liquide (20).
  12. Procédé selon la revendication 1, dans lequel le récipient (2) est fourni sous la forme d'un récipient à bas prix et jetable, et dans lequel il est jeté après le traitement des gouttelettes de liquide (19) sur la surface supérieure hydrophobe (16) du film polymère plat (14) par électromouillage commandé par l'instrument de manipulation de gouttelette de liquide (20).
EP13770655.2A 2013-03-04 2013-09-23 Manipulation de la taille des gouttes liquides en microfluidique numérique Active EP2931425B1 (fr)

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US13/784,168 US8936708B2 (en) 2008-12-17 2013-03-04 Manipulating the size of liquid droplets in digital microfluidics
PCT/EP2013/069703 WO2014135232A1 (fr) 2013-03-04 2013-09-23 Manipulation de la taille de gouttelettes de liquide en microfluidique numérique

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