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WO2025181490A1 - Améliorations apportées à ou se rapportant à un dispositif de distribution - Google Patents

Améliorations apportées à ou se rapportant à un dispositif de distribution

Info

Publication number
WO2025181490A1
WO2025181490A1 PCT/GB2025/050403 GB2025050403W WO2025181490A1 WO 2025181490 A1 WO2025181490 A1 WO 2025181490A1 GB 2025050403 W GB2025050403 W GB 2025050403W WO 2025181490 A1 WO2025181490 A1 WO 2025181490A1
Authority
WO
WIPO (PCT)
Prior art keywords
chip
micro
entities
carrier fluid
microdroplets
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/GB2025/050403
Other languages
English (en)
Inventor
Thomas Henry ISAAC
Gareth William Harry EVANS
Robert Wootton
Richard Jeremy INGHAM
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Lightcast Discovery Ltd
Original Assignee
Lightcast Discovery Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Lightcast Discovery Ltd filed Critical Lightcast Discovery Ltd
Publication of WO2025181490A1 publication Critical patent/WO2025181490A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—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
    • B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • 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
    • 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
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • 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
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • 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
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0469—Buoyancy
    • 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
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5088—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above confining liquids at a location by surface tension, e.g. virtual wells on plates, wires

Definitions

  • the present invention relates to improvements in or relating to a chip on which a plurality of micro-entities can be tracked, manipulated and analysed. Movement of the micro-entities on the chip may be actuated magnetically, electrokinetically or even acoustically.
  • the present invention may be implemented with an electrowetting-on-dielectric (EWOD) or optoelectrowetting-on-dielectric (oEWOD) chip and the micro-entities may be a plurality of microdroplets.
  • EWOD electrowetting-on-dielectric
  • oEWOD optoelectrowetting-on-dielectric
  • EWOD electrowetting- on-dielectric
  • oEWOD optoelectrowetting-on-dielectric
  • a chip for manipulation of a plurality of micro-entities comprising: a microfluidic space configured to accommodate a plurality of micro-entities in a substantially planar array; wherein the height of the microfluidic space in the plane perpendicular to the array is sized to accommodate a single layer of micro-entities; wherein the microfluidic space contains a carrier fluid in which the array of micro-entities can be localised and manipulated using a force; wherein the microfluidic space has an exit orifice that extends a distance in the plane of the array that is at least double the height of the microfluidic space so that two or more micro-entities in parallel can be moved through the exit orifice simultaneously under the force; and a dispense region configured to provide a flow of carrier fluid substantially non-parallel to the exit orifice so that the micro-entities are moved by the carrier fluid through the dispense region and out of the chip.
  • a chip for manipulation of a plurality of micro-entities comprising: a microfluidic space containing a carrier fluid in which an array of micro-entities can be localised and manipulated using a force; wherein the microfluidic space has an exit orifice sized to accommodate more than two micro-entities in parallel moved under the force; a dispense region configured to provide a flow of carrier fluid substantially non-parallel to the exit orifice so that the micro-entities are moved by the carrier fluid through the dispense region and out of the chip.
  • the chip can be an electrowetting, EWOD or oEWOD chip.
  • the chip can be an optical tweezer device, or an opto-electronic tweezer (OET) device, or a dielectrophoresis (DEP) device.
  • Electrowetting, EWOD, oEWOD, Optical tweezers, opto-electronic tweezers and dielectrophoresis (DEP), and the like, are differing techniques which can be utilised as appropriate for the particular application of interest.
  • an EWOD or oEWOD chip for manipulation of a plurality of microdroplets, the chip comprising: a microfluidic space containing a carrier fluid in which an array of microdroplets can be localised and manipulated using EWOD or oEWOD; wherein the microfluidic space has an exit orifice sized to accommodate more than two microdroplets in parallel moved under EWOD or oEWOD; a dispense region configured to provide a flow of carrier fluid substantially non-parallel to the exit orifice so that the microdroplets are moved by the carrier fluid through the dispense region and out of the chip.
  • the device as disclosed herein enables the efficient removal of unwanted microdroplets and/or micro-entities from a microfluidic device.
  • the unwanted microdroplets can be quickly removed from the chip.
  • This increases the capacity within the chip for manipulating microdroplets and/or microentities of interest.
  • moving multiple microdroplets and/or micro-entities out of the chip simultaneously reduces or prevents a backlog of unwanted microdroplets and/or micro-entities within the chip and thus, it can minimise the loading time of microdroplets into the chip.
  • This can be advantageous as it enables experiments to be quickly and efficiently carried out on the chip and reduces experimental downtime for the user.
  • the chip of the present invention provides a simplified and cost- effective approach for dispensing multiple microdroplets and/or micro-entities out of the chip.
  • the dispense region may be shaped and configured in order to minimise the flow of carrier fluid into the microfluidic space.
  • Carrier fluid flow at a high velocity rate can interfere with the oEWOD or EWOD forces applied to microdroplets and therefore, this may disrupt or disturb the microdroplets being held by EWOD or oEWOD within the array.
  • the user is able to apply EWOD or oEWOD manipulation of microdroplets and/or micro-entities with greater control and efficiency within the microfluidic space.
  • the chip of the present invention is configured to be capable of the manipulation of tens, hundreds, thousands and/or millions of microdroplets and/or micro-entities, simultaneously using a force such as an (opto)electrowetting force.
  • the micro-entities may be contained within a microdroplet that encapsulates one or more micro-entity and moves with that which it encapsulates.
  • the micro-entities may be contained within a micro-object, which is a stationary physical construct, such as a sequestration pen.
  • the dispense region can be shaped and configured in order to maximise the rate at which micro-entities, either independently or encapsulated within microdroplets, can be moved through the dispense region and out of the chip.
  • a carrier fluid is provided via one or more carrier inlet(s).
  • the carrier inlets are shaped and configured to enable the carrier fluid flow to be directed towards an outlet of the dispense region.
  • the carrier fluid provided within the dispense region is at a velocity that is more than zero i.e. at a higher velocity than the velocity of the carrier fluid provided within the microfluidic space.
  • the carrier fluid may have a centreline velocity of at least 50pm/s, typically 500pm/s, as high as 2000pm/s and in an extreme case up to 2cm/s. Micro-entities are moved by the high- velocity carrier fluid flow in the direction towards an outlet of the dispense region and out of the chip.
  • the carrier fluid can be oil, HFE 7500 and/or media.
  • the carrier fluid can be a fluorocarbon fluid such as FC40, HFE7700, HFE 7100, Opteon SF10, Opteon SF20.
  • the carrier fluid can be a mineral oil, paraffin oil, any suitable hydrocarbon oils, a vegetable oil or fat; or any other suitable electrically-insulating liquid.
  • the chip as disclosed herein can be further advantageous because less carrier fluid e.g. oil needs to be consumed compared to other known chips in the art in order to move microdroplets out of the chip.
  • the chip of the present invention uses much less valuable resources such as the oil carrier fluid and thus, many workflows using the chip of the present invention are more practical, sustainable and cost-effective.
  • the carrier fluid may be water or a water-based medium such as a buffer, electrolyte solution or cell media and the micro-entities may be dispersed within this water based medium.
  • the carrier fluid may be a gas.
  • the exit orifice may be sized to accommodate at least 5 microdroplets in parallel. In some embodiments, the exit orifice may be sized to accommodate at least 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more microdroplets in parallel.
  • the wide structure of the exit orifice allows for several parallel trains of micro-entities to enter the structure unimpeded. Preferably, there can be between 1 to 50 parallel trains of microdroplets entering the exit orifice at the same time.
  • This configuration provides an exit orifice with a droplet-offloading capacity at least as high as the capacity of the inlet loading structures for the chip. It allows micro-entities to be supplied from a highly parallel storage array without a time-consuming rearrangement to enable sequential removal. This makes chip re-use more practical.
  • the exit orifice may comprise a single gap which makes optimal use of the space and provides simplicity of manufacture and alignment.
  • the exit orifice may be configured from a series of gaps, a sieve, a semi-permeable membrane, such as a pillar, to enable the microdroplet and/or the micro-entity to enter the dispense region of the chip.
  • the movement of microdroplets and/or micro-entities through the dispense region and out of the chip is flow-driven via a carrier fluid at a high velocity.
  • the dispense region may comprise a cavity with a depth of between 100pm to 200pm, or it may be more than 100, 110, 120, 130, 140, 150, 160, 170, 180 or 190 pm. In some embodiments, the dispense region may comprise a cavity with a depth of less than 200, 190, 180, 170, 160, 150, 140, 130, 120 or 110 pm.
  • the movement of microdroplets and/or micro-entities through the dispense region and out of the chip can be buoyancy-driven.
  • the microdroplets and/or micro-entities can be driven to the edge of the cavity and float, under their buoyancy in the carrier fluid flow, into the cavity and out towards the outlet of the dispense region.
  • This embodiment of the invention has an advantage in that the buoyant force acts to propel the microdroplets and/or micro-entities out of the chip more quickly than flow alone.
  • microdroplets and/or micro-entities through the dispense region and out of the chip can be buoyancy-driven requiring less oil to be consumed in order to purge microdroplets and/or micro-entities from the chip.
  • the force can be, but is not limited to, an optically-mediated or a non-optically-mediated force.
  • the force can be dielectrophoresis (DEP), EWOD and/or oEWOD.
  • DEP dielectrophoresis
  • EWOD EWOD
  • oEWOD oEWOD
  • the microdroplet which may contain one or more micro-entities can be moved through the exit orifice and into the dispense region of the chip.
  • empty microdroplets i.e. microdroplets that do not contain a micro-entity can be moved through the exit orifice using a force, such as DEP, EWOD or oEWOD (as appropriate) and into the dispense region of the chip.
  • the sequestration pen may contain one or more micro-entities.
  • a plurality of sequestration pens is located within the microfluidic space.
  • the contents of the sequestration pen i.e. the micro-entity or entities contained within the sequestration pen can be moved out of the pen and through the exit orifice and into the dispense region under a force, such as an optically-mediated force or non-optically-mediated force.
  • the micro-entity or micro-entities can be one or more of the following: a microdroplet; a cell, a part of a cell and/or a bead such as a microbead, an organelle, an organism, a liposome or a biosynthetic structure, a micro-capsule, a gel bead, a tentagel bead or a vesicle. Where more than one micro-entity is provided, the micro-entities may be of the same type or differing types.
  • micro-entities may be agglomerated together in any appropriate manner including, but not limited to a micro-entity adhered to the surface of another micro-entity such as a cell attached to the surface of a bead; a micro-entity encapsulated in another micro-entity such as a cell, organelle or vesicle contained within a microdroplet.
  • the methods may comprise targeted cell-cell interactions where two or more selected cells are brought together. It may be desirable to analyse any kind of cell using the methods of the present invention, but the cells may be of the same type, for example they are B cells or T cells (lymphocytes).
  • the cell(s) may be natural or it may be artificial.
  • the cell(s) may be microcells.
  • the methods of the invention may be cell free and use part(s) of a cell(s), for example nuclei and/or mitohbrichondria.
  • the cell may be a cell from a human or animal, optionally a mammal, a plant cell, insect cell, fungal cell, bacterial cell, ameobal cell, a yeast, macrophage or hybridoma, and are selected from, but are not limited to: CHO, Jurkat, CAMA, HeLa, B-cell, T-cell, MCF-7, MDAMB-231 , E. coli and Salmonella.
  • the cell may be a cell-fusion such as a hybridoma.
  • the cells may be taken from a cell culture, for example a culture of stem cells, pluripotent cells, genetically engineered cells and the like.
  • micro-entity e.g. a cell is derived from a sample
  • this may be any human, animal, environmental (natural, contrived or modified), or food sample containing at least one micro-entity type e.g. cell type.
  • the sample may be selected from: stool, peripheral blood, sera, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humour, amniotic fluid, cerumen, breast milk, broncheoalveolar lavage fluid, semen, prostatic fluid, cowper's fluid or pre-ejaculatory fluid, female ejaculate, sweat, faecal matter, hair, tears, cyst fluid, pleural and peritoneal fluid, pericardial fluid, lymph, chyme, chyle, bile, interstitial fluid, menses, pus, sebum, vomit, vaginal secretions, mammary secretions, mucosal secretion
  • the cell may be isolated from a patient or individual.
  • the chip of the present invention as described herein may be used to screen such cells and return them to the patient (autologous cell transfer).
  • the cells may be isolated from one individual and selected to be administered to a patient (allogenic cell transfer).
  • the cell may be an immune system cell.
  • Such cells include monocytes, macrophages, osteoclasts, neutrophils (polymorphonuclear leukocytes) dendritic cells, microglial cells, mast cells, T cells (including helper T cells, regulatory T cells, cytotoxic T cells and natural killer T cells), B cells, natural killer cells and hematopoietic stem cells.
  • the cell can be a CHO cell or it can be a Jurkat cell.
  • Chinese hamster ovary (CHO) cells are modified to produce an immuno-therapeutic drug (e.g. a TCR) and then emulsified into microdroplets and loaded onto the microfluidic platform. Empty or multi-occupancy microdroplets are discarded. The remaining microdroplets containing single CHO cells are incubated on-chip to promote the production of the immunotherapeutic drug. The CHO containing droplets may then be split to obtain multiple doses of the drug produced by each cell.
  • T cells and target tumour cells are separately emulsified and loaded into arrays onto the microfluidic platform, adjusting the cell occupancy of each microdroplet as desired.
  • T cell and tumour cell arrays are then merged.
  • a second merge operation is used to add a dose of immunotherapeutic drug, tracking which CHO cell each dose came from.
  • the resulting assay is incubated and T-cell killing behaviour is monitored using the detection system by detecting caspase 3/7 fluorescence, a fluorescent marker of apoptosis.
  • CHO cells that produced effective doses of the test drug can be dispensed from the device into a well plate.
  • the cell may be a pluripotent or stem cell, isolated or prepared via culturing techniques.
  • the pluripotent stem cells may be reprogrammed mature cell types.
  • the cell may be genetically engineered prior to encapsulation into the microdroplet.
  • the cell may be genetically engineered after encapsulation in the microdroplet.
  • Genetic engineering of the cell may be by any suitable method, including transduction (viral gene transfer), gene editing (using a nuclease such as zinc finger nucleases, TALEN, CRISPR/Cas9 base and prime editing) non-viral gene delivery (such as nanoparticle delivery), gene knock down, gene knock in and gene manipulation using RNA, for example, such as gene silencing or activation, or optogenetics.
  • transduction viral gene transfer
  • gene editing using a nuclease such as zinc finger nucleases, TALEN, CRISPR/Cas9 base and prime editing
  • non-viral gene delivery such as nanoparticle delivery
  • gene knock down gene knock in and gene manipulation using RNA, for example, such as gene silencing or activation, or optogenetics.
  • the genetic engineering generally involves the introduction of a genetic element into the cell, by any suitable means.
  • the biological and/or chemical entity is any one or more of the following entities: an antibody; an antigen; a receptor; a substrate; an enzyme; a ligand; a nucleic acid; a cell; a part of a cell; an extracellular vesicle; a liposome; a polymer; a chemical; a drug; a FRET reporter; a chemiluminescent material; a sample of tissue; a virus or bacteriophage; a cytokine; and/or a protein.
  • an EWOD or oEWOD chip according to the previous aspect, comprising a plurality of dispense regions.
  • the microfluidic space may comprise a single exit orifice leading into a plurality of dispense regions.
  • the exit orifice may be divided into multiple sub-regions. The exit orifice can be sized to accommodate more than two microdroplets in parallel moving into the plurality of dispense regions under EWOD or oEWOD control.
  • the number of dispense regions provided in one chip can be adjusted for example, by increasing or decreasing the number of dispense regions provided on the chip in order to ensure that there is sufficient capacity to remove microdroplets out of the chip at an efficient rate.
  • the species may be chemical, biochemical, or biological in nature and may be of any size.
  • the micro-entity is a species of micro dimension.
  • the present invention may provide an agonist/antagonist to a micro-entity as identified by the screening, selection and/or isolation method disclosed herein.
  • the chip, apparatus, method or species as disclosed herein there is provided a use of the chip, apparatus, method or species as disclosed herein. According to an aspect of the present invention, there is provided a use of the chip, apparatus, method or species as disclosed herein in therapy.
  • the present invention may provide for a use of the chip, apparatus, method or species as disclosed herein in making a product.
  • the product made may be chemical, biochemical, or biological in nature.
  • the use may be peptide synthesis.
  • the use may be synthetic biology.
  • the use may be cell line engineering or development.
  • the use may be cell therapy.
  • the use may be drug discovery.
  • the analysis may be physical, chemical, or biological.
  • the use may be sub-cellular imaging.
  • the use may be high content imaging.
  • the use may be diagnostics.
  • the use may be a biological assay.
  • the biological assay may be high throughput screening.
  • the biological assay may be ELISA.
  • the use may be cell secretion.
  • the use may be QC safety profiling.
  • Figure 1 shows a chip, such as an EWOD or oEWOD chip, according to the present invention
  • Figure 3 shows an alternative embodiment of the EWOD or oEWOD chip according to the present invention
  • Figure 4 provides a side view of the EWOD or oEWOD chip according to the present invention.
  • Figure 5 shows an embodiment of the chip containing a plurality of dispense regions
  • a chip 10 such as an EWOD or oEWOD chip, comprising a microfluidic space 12 containing a carrier fluid in which an array of microdroplets and/or an array of micro-entities, such as cells, can be localised and manipulated using a force, for example EWOD or oEWOD forces (as appropriate).
  • the micro-entities may be contained within a microdroplet that encapsulates one or more micro-entity and moves with that which it encapsulates.
  • the microentity can be encapsulated within the micro-object.
  • a cell can be contained within a microdroplet and/or it can be contained within a sequestration pen.
  • the microfluidic space 12 has an exit orifice 14 that is sized to accommodate more than two microdroplets in parallel moved under EWOD or oEWOD. In some instances, five or more microdroplets in parallel can be moved through the exit orifice 14 under EWOD or oEWOD.
  • the chip 10 further comprises a dispense region 16, which is configured to provide a flow of carrier fluid substantially non-parallel to the exit office 14. The microdroplets within the dispense region 16 are released from EWOD or oEWOD control.
  • the high flow velocity of the carrier fluid provided within the dispense region 16 is configured to move the plurality of microdroplets through the dispense region 16 and out of the chip 10.
  • a carrier inlet channel 26 can be provided on either side of the dispense region 16.
  • the carrier inlet channel 26 comprises an inlet port 25 and is configured to introduce a carrier fluid flow into the dispense region 16.
  • an EWOD or oEWOD chip 10 comprising a microfluidic space 12, an exit orifice 14 and a dispense region 16.
  • the microfluidic space receives a plurality of microdroplets 18.
  • the microdroplets 18 can be arranged in an array within the microfluidic space 12 as shown in Figure 2. At least a subset of the microdroplets 18 may undergo one or more of the following EWOD or oEWOD operations or manipulation including: holding, merging, splitting or discarding at least a subset of microdroplets in an array and/or an entire array of microdroplets. In some cases, a subset of microdroplets in the array or an entire array of microdroplets may be held stationary under EWOD or oEWOD forces.
  • EWOD or oEWOD operation may include sequential merging of microdroplets. For example, to augment at least a subset of microdroplets in the array with further microdroplets that may contain micro-entities or other species.
  • the carrier fluid flow within the microfluidic space 12 of the chip 10 is substantially at zero velocity.
  • At least a subset of microdroplets 18 within an array in the microfluidic space 12 may be selected and moved towards the exit orifice 14, as indicated by the arrow 20 in Figure 2.
  • the exit orifice 14 has a width that is sufficiently wide in the vertical and/or horizontal direction to accommodate two or more microdroplets 18 moving through the exit orifice 14 simultaneously under EWOD or oEWOD.
  • the exit orifice 14 has a sufficient width to accommodate up to one hundred microdroplets 18 in parallel.
  • the exit orifice 14 can accommodate between 2 to 5, 2 to 10, 2 to 20, 2 to 50 or 2 to 60, 2 to 75, 2 to 80 or 2 to 90 microdroplets 18 in parallel.
  • the exit orifice 14 can accommodate 30 microdroplets in parallel. In some instances, the exit orifice 14 has a sufficient width to accommodate at least 10 microdroplets in parallel.
  • the microdroplets 18 may be released from EWOD or oEWOD control as they enter the dispense region 16.
  • the exit orifice 14 enables the constant and controlled delivery of microdroplets under EWOD or oEWOD control into the dispense region 16.
  • the width of the exit orifice 14 is important because it allows for several parallel trains of microdroplets 18 to enter unimpeded. As shown in Figure 2, an area in the dispense region 16 contains a high-velocity carrier phase flow directed towards an outlet 24 in the chip 10.
  • the dispense region 16 as disclosed in the present invention has an optimal shape and orientation such that there is a near-zero component of flow directed towards the exit orifice 14.
  • the angle of the carrier phase inlets 26, shown in Figure 2 is optimised such that the carrier fluid flows are introduced into the dispense region 16 in the direction towards the outlet 24.
  • the high velocity carrier fluid will carry the microdroplets 18 within the dispense region 16 towards the outlet 24 and out of the chip 10.
  • the angle a of the inlets 26 for introducing the carrier fluid flows into the dispense region 16 is between 60 to 70 degrees with respect to the exit orifice 14. Furthermore, the width of the carrier phase inlets 26 is optimally between 100pm and 200pm. The width of the inlets 26 may determine the peak velocity at the edge of the dispense region 16.
  • FIG. 3 there is shown an alternative embodiment of an EWOD or oEWOD chip 10 according to the present invention.
  • Figure 3 shows the EWOD or oEWOD chip 10 comprising a microfluidic space 12, an exit orifice 14 and a dispense region 16.
  • the dispense region 16 can comprise a substrate (not shown in the accompanying drawing) in which a cavity 28 is provided on top of the substrate.
  • the cavity 28 can have a depth of between 100 to 200pm.
  • the cavity 28 can be an approximately 150pm deep space and ablated into the substrate, such as a glass substrate, of the EWOD or oEWOD chip 10.
  • microdroplets 18 can be driven to the edge of the cavity 28 of the dispense region 16 and float under their buoyancy in the carrier fluid flow into the cavity 28. As shown in Figures 3 and 4, the carrier fluid flow enters the dispense region 16 through the carrier inlets 26 to move the microdroplets out of the chip 10 via an outlet 24.
  • the outlet 24 further comprises a conduit 30, such as a tube to control the direction of microdroplets 18 transportation out of the EWOD or oEWOD chip 10.
  • the microdroplet 18 can exit the chip 10 via a hole 31 before entering the conduit 30.
  • the carrier phase inlets 26 comprises an opening32 to introduce a carrier phase into the channel 26.
  • the chip 10 can comprise more than one layer.
  • the chip may be made of multi-layers or it can be made from a single layer.
  • the chip 10 can be made from a first layer 33 and a second layer 34.
  • the first and second layers 33, 34 can be made from the same material or they can be made from different materials.
  • the configuration shown in Figures 3 and 4 has the advantage that the buoyant force acts to propel microdroplets 18 out of the chip 10 more quickly than flow alone. Furthermore, less oil carrier fluid flow is required to move the microdroplets 18 out of the chip 10.
  • a chip 10 such as an EWOD or oEWOD chip, comprising a microfluidic space 12, an exit orifice 14 and a plurality of dispense regions 16.
  • Each dispense region 16 has two symmetrical inlets for introducing a carrier fluid flow into the dispense region, such that micro-entities which may or may not be encapsulated in microdroplets are moved out of the chip under flow of the carrier fluid.
  • the arrangement of the plurality of dispense regions 16 provided on the chip 10, as shown in Figure 5, can be particularly useful for moving the micro-entities out of the chip 10 at high speed.
  • a chip 10 such as an EWOD or oEWOD chip, comprising a microfluidic space 12, an exit orifice 14 and a plurality of dispense regions 16.
  • the dispense regions 16 comprise a cavity 28.
  • the cavity 28 can have a depth of between 100 to 200pm.
  • a micro-entity which may or may not be encapsulated in a microdroplet, can be driven to the edge of the cavity 28 of the dispense region 16 and float, as a result of their buoyancy in the carrier fluid flow, into the cavity 28.
  • a carrier fluid can then be subsequently introduced into the dispense region via the two inlets provided on either side of the dispense region 16 to drive the micro entities out of the dispense region through an outlet.
  • the arrangement of the plurality of dispense regions 16 provided on the chip 10, as shown in Figures 6A and 6B, can be particularly useful for moving the micro-entities out of the chip 10 in a high throughput manner.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Fluid Mechanics (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)

Abstract

L'invention concerne une puce destinée à la manipulation d'une pluralité de micro-entités, la puce comprenant : un espace microfluidique conçu pour loger une pluralité de micro-entités dans un réseau sensiblement plan ; la hauteur de l'espace microfluidique dans le plan perpendiculaire au réseau étant dimensionnée pour loger une seule couche de micro-entités ; l'espace microfluidique contenant un fluide porteur dans lequel le réseau de micro-entités peut être localisé et manipulé au moyen d'une force ; l'espace microfluidique comportant un orifice de sortie qui s'étend sur une distance dans le plan du réseau qui est au moins le double de la hauteur de l'espace microfluidique de sorte que deux micro-entités ou plus en parallèle puissent être déplacées à travers l'orifice de sortie simultanément sous la force ; et une région de distribution conçue pour fournir un écoulement de fluide porteur sensiblement non parallèle à l'orifice de sortie de sorte que les micro-entités soient déplacées par le fluide porteur à travers la région de distribution et hors de la puce.
PCT/GB2025/050403 2024-03-01 2025-02-28 Améliorations apportées à ou se rapportant à un dispositif de distribution Pending WO2025181490A1 (fr)

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GB2403032.2A GB2638777A (en) 2024-03-01 2024-03-01 Improvements in or relating to a dispensing device
GB2403032.2 2024-03-01

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WO2025181490A1 true WO2025181490A1 (fr) 2025-09-04

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Citations (4)

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Publication number Priority date Publication date Assignee Title
WO2015076251A1 (fr) * 2013-11-21 2015-05-28 国立大学法人東京大学 Appareil de sélection et d'élimination de cellules, et procédé de sélection et d'élimination de cellules
KR20160002117U (ko) * 2016-05-10 2016-06-20 서울대학교산학협력단 다중 액적 생성 장치
US20200055052A1 (en) * 2016-10-11 2020-02-20 The Regents Of The University Of California Systems and methods to encapsulate and preserve organic matter for analysis
AU2021281604A1 (en) * 2020-05-28 2022-12-22 Lightcast Discovery Ltd Improvements in or relating to a device and method for dispensing a droplet

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Publication number Priority date Publication date Assignee Title
WO2020176882A1 (fr) * 2019-02-28 2020-09-03 10X Genomics, Inc. Dispositifs, systèmes et procédés pour augmenter l'efficacité de formation de gouttelettes
CZ20207A3 (cs) * 2020-01-06 2021-03-31 Otakar Černý Způsob stanovení spotřeby energie na vytápění nebo chlazení
CN116171200A (zh) * 2020-09-02 2023-05-26 10X基因组学有限公司 用于高通量液滴形成的装置、系统和方法
CA3197695A1 (fr) * 2020-10-05 2022-04-14 Lightcast Discovery Ltd Ameliorations apportees ou relatives a un dispositif et procedes pour faciliter la manipulation de microgouttelettes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015076251A1 (fr) * 2013-11-21 2015-05-28 国立大学法人東京大学 Appareil de sélection et d'élimination de cellules, et procédé de sélection et d'élimination de cellules
KR20160002117U (ko) * 2016-05-10 2016-06-20 서울대학교산학협력단 다중 액적 생성 장치
US20200055052A1 (en) * 2016-10-11 2020-02-20 The Regents Of The University Of California Systems and methods to encapsulate and preserve organic matter for analysis
AU2021281604A1 (en) * 2020-05-28 2022-12-22 Lightcast Discovery Ltd Improvements in or relating to a device and method for dispensing a droplet

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GB202403032D0 (en) 2024-04-17

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