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WO2022086565A1 - Microfluidique ayant des épaisseurs de gouttelettes réduites - Google Patents

Microfluidique ayant des épaisseurs de gouttelettes réduites Download PDF

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Publication number
WO2022086565A1
WO2022086565A1 PCT/US2020/057181 US2020057181W WO2022086565A1 WO 2022086565 A1 WO2022086565 A1 WO 2022086565A1 US 2020057181 W US2020057181 W US 2020057181W WO 2022086565 A1 WO2022086565 A1 WO 2022086565A1
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WO
WIPO (PCT)
Prior art keywords
site
array
gap
droplet
sites
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.)
Ceased
Application number
PCT/US2020/057181
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English (en)
Inventor
Michael W. Cumbie
Viktor Shkolnikov
Carson DENISON
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.)
Hewlett Packard Development Co LP
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Hewlett Packard Development Co LP
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Priority to PCT/US2020/057181 priority Critical patent/WO2022086565A1/fr
Publication of WO2022086565A1 publication Critical patent/WO2022086565A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—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
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • 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
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • 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
    • B01L2200/0673—Handling of plugs of fluid surrounded by immiscible fluid
    • 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
    • B01L2300/0887—Laminated structure
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1822—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using Peltier elements
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1827—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1894—Cooling means; Cryo cooling
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • 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
    • B01L2400/0424—Dielectrophoretic 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

Definitions

  • Microfluidic devices may be used to perform a variety of chemical, biological, and biochemical processes, such as nucleic acid amplification.
  • a microfluidic device may be capable of heating a fluid as part of the process implemented by the device.
  • FIG. 1 is a cross-sectional view, along section line A-A of FIG. 2, of an example device to reduce a thickness of a fluid droplet during a temperature change.
  • FIG. 2 is a plan view of the example device of FIG. 1 , showing an array of electrodes including an electrode located at a temperature change site with reduced thickness.
  • FIG. 3 is a schematic view of an example device to reduce a thickness of a fluid droplet during a temperature change with multiple electrodes located at a temperature change site with reduced thickness.
  • FIG. 4 is a schematic view of an example device to reduce a thickness of a fluid droplet during a temperature change, in which a reduced- thickness temperature change site has area greater than another site.
  • FIG. 5 is a schematic view of an example device for a nucleic acid amplification process, wherein an array of sites includes some sites located at positions with reduced gap thickness to facilitate a temperature change.
  • FIG. 6 is a cross-sectional view of the example device along section line B-B of FIG. 5.
  • FIG. 7 is a flowchart of an example method of transporting a droplet to a narrowed site between bodies and applying a thermal bias to a droplet at the site.
  • FIG. 8 is a flowchart of an example method of transporting a droplet among various sites of to provide various thicknesses to the droplet and applying a thermal bias at various sites.
  • FIG. 9 is a graph of example times to reach target temperature for thick and thin droplets.
  • an electrode array may be used to transport microdroplets through an array of sites.
  • Sites may be used for reagent mixing, transport, heating, and so on.
  • Sites are normally defined by a pair of planar bodies that are separated by a uniform gap.
  • a heater may be disposed at one of the bodies. When heating is desired, a microdroplet is transported to a site in the effective range of the heater.
  • Heating or cooling can be performed more quickly if the gap is reduced at the location of the thermal element (e.g., heater or cooling device) to provide a thinned microdroplet. That is, a localized reduced gap may force a droplet to thin in proximity to a thermal element. The temperature of a thinned microdroplet may be changed in less time, which allows for a reduction of an overall time of a process implemented by a digital microfluidic device.
  • the thermal element e.g., heater or cooling device
  • Such an arrangement may be used to implement a polymerase chain reaction process, such as a reverse transcription polymerase chain reaction (RT-PCR) process, real-time or quantitative (qPCR) process, or similar.
  • RT-PCR reverse transcription polymerase chain reaction
  • qPCR quantitative
  • thermal cycle time and thus total time for a PGR or similar process may be reduced by way of thinning droplets when applying (or removing) heat.
  • a normal or wider gap may be provided to allow for effective fluorescence measurements.
  • An example device includes a pair of separated bodies to contain microdroplets therebetween and an array of electrodes to define an array of sites between the pair of separated bodies.
  • An electrode of the array of electrodes is controllable to position a microdroplet at a corresponding site of the array of sites. A distance between the pair of separated bodies varies among the array of sites.
  • Another example device includes a substrate and a cover separated from the substrate by a gap that includes a narrowed portion.
  • the gap constrains a thickness of a fluid droplet situated between the substrate and the cover.
  • the device further includes an array of electrodes at the substrate. The array of electrodes is to move the fluid droplet within the gap in response to a charge applied to an electrode of the array of electrodes.
  • the device further includes a thermal element positioned with respect to the substrate to change a temperature of the fluid droplet. The thermal element is positioned at the narrowed portion of the gap to reduce the thickness of the fluid droplet while undergoing the change in temperature.
  • the array of electrodes may include a first electrode at a first site.
  • the first electrode may draw the fluid droplet to the first site.
  • the array of electrodes may further include a second electrode at a second site.
  • the second electrode may draw the fluid droplet to the second site.
  • the second site may be located at the narrowed portion of the gap.
  • the second site may be about equal in area to the first site.
  • the second site may be larger in area than the first site.
  • the first and second sites may define fluid volumes that are approximately equal.
  • the array of electrodes may include a plurality of the second electrodes at a plurality of contiguous second sites.
  • the narrowed portion of the gap may extend throughout the plurality of contiguous second sites.
  • the cover may include a transparent or semi-transparent material that includes a thickened portion facing the substrate. The thickened portion may define the narrowed portion of the gap.
  • the gap may be between about 0.2 mm and 0.5 mm.
  • the narrowed portion of the gap may be between about 0.1 mm and 0.25 mm.
  • FIG. 1 shows an example device 100 to apply a temperature change (e.g., heat or cool) to a fluid droplet while reducing the thickness of the droplet.
  • the device 100 may be used in a chemical, biological, or biochemical process, such as a PGR process or similar nucleic acid amplification process.
  • the device 100 may be provided with a nucleic acid sample as well as reagents and materials to perform a PGR process.
  • the device 100 may be connected to a source of electrical power and a controller to control the device 100 and capture measurements from the device 100 to carry out the process.
  • Fluid droplets may be moved conveyed and heated or cooled by the device to perform the process, and such droplets may be referred to as microdroplets with an examples of suitable volumes being on the order of pL, nL, pL, and fL.
  • the device 100 includes a pair of separated bodies, such as a substrate 102 and a cover 104, to contain microdroplets (shown in dashed line) at an array of sites between the bodies.
  • the substrate 102 and cover 104 may be generally planar bodies.
  • the substrate 102 may include a printed circuit board (PCB), glass, a dielectric, or similar material.
  • a non-wetting or hydrophobic coating such as an immiscible fluorocarbon oil (e.g., FluorinertTM FC-40), may be provided to the substrate 102.
  • the cover 104 may include transparent or semi-transparent material, such as a polymer (e.g., polyethylene terephthalate or PET) and/or a transparent metallic layer (e.g., indium tin oxide or ITO), to allow optical measurements to be captured, such as by fluorescence generated by a markers or other constituent of a PCR process.
  • a transparent or semi-transparent material such as a polymer (e.g., polyethylene terephthalate or PET) and/or a transparent metallic layer (e.g., indium tin oxide or ITO), to allow optical measurements to be captured, such as by fluorescence generated by a markers or other constituent of a PCR process.
  • a non-wetting or hydrophobic coating may be provided to the cover 104.
  • the substrate 102 and cover 104 are separated by a gap 106.
  • the gap 106 is a distance that constrains the thicknesses of the microdroplets.
  • the cover 104 may include a thickened portion 108 facing the substrate 102, and the thickened portion 108 may define the narrowed portion 1 10 of the gap 106.
  • the size of the gap 106 may be varied according to site.
  • the gap 106 may include a narrowed portion 1 10 at a site where a thermal bias is applied to microdroplets, so that the temperature change takes less time.
  • the device 100 further includes an array of electrodes 1 12.
  • the electrodes 112 may be energized and deenergized to move microdroplets among the sites within the gap 106. Droplets may be moved from an uncharged electrode to a charged electrode, or vice versa, so as to be continuously transported via the electrodes by sequentially energizing/deenergizing the electrodes For example, to move a droplet into the narrowed portion 1 10 of the gap 106, a first electrode 116 outside the narrowed portion 110 may be deenergized and a second electrode 1 18 aligned with the narrowed portion 110 may be energized.
  • the electrode array 1 12 may be arranged to define a desired arrangement of sites between the substrate 102 and cover 104, so that microdroplets may be moved among the sites to undergo various acts of the process performed by the device 100.
  • the device 100 further includes a thermal element 114 to positioned with respect to the substrate 102 to change a temperature of a microdroplet located at the site of the thermal element 114.
  • a thermal element 114 may additional or alternatively be disposed on the cover 104.
  • the thermal element 114 is positioned at the narrowed portion 1 10 of the gap 106 so that the temperature change is performed on microdroplets with reduced thickness as constrained by the narrowed portion 110 of the gap 106. Reducing the thickness of the microdroplet reduces the time it takes to heat or cool the microdroplet to a temperature desired for the performance of the process.
  • the thermal element 114 may include a heater, such as a heater formed by a resistive heating layer disposed on or in the substrate 102.
  • the thermal element 114 may include a cooling device, whether active or passive, such as a body of highly thermally conductive material (e.g. copper), a heat pipe, a cooling channel for liquid coolant flow, a thermoelectric or Peltier device, or similar.
  • a cooling device whether active or passive, such as a body of highly thermally conductive material (e.g. copper), a heat pipe, a cooling channel for liquid coolant flow, a thermoelectric or Peltier device, or similar.
  • the time to change the temperature of a microdroplet may be proportional to the square of the governing dimension, which in the examples discussed herein is the thickness of the microdroplet as constrained by the separation of two generally planar bodies.
  • droplet velocity through a thinned region may increase to maintain an average constant flowrate.
  • velocity increase is linear with respect to constrained thickness.
  • a droplet whose thickness is reduced by half has its volumetric flow slowed by half, but has its heating time reduced fourfold.
  • heating time is still reduced by half, in that the droplet moves half as fast but is heated four times as quickly.
  • the gap 106 may be selected to be between about a distance of 0.2 mm and about a distance of 0.5 mm.
  • the narrowed portion 110 of the gap 106 may be selected to be between about a distance of 0.1 mm and about a distance of 0.25 mm.
  • These dimensions are merely examples, which provide droplet thickness reduction factors in the range of about 0.2 (/.e., thickness reduced to about 20% of original) to about 0.75 or more (/.e., thickness reduced to about 75% of original). It should be understood that as droplet thickness is reduced, the area occupied by the droplet (in a plane perpendicular to its thickness) increases because of conservation of volume (or mass). Flow resistance may also increase as droplet thickness decreases due to phenomena such as surface tension.
  • thickness reduction may be selected from very little reduction (e.g., 0.8-0.95 of original) to very high reduction (e.g., 0.1 -0.2 of original) based on the process being performed and other considerations, such as the resulting increased droplet area and possible increase in flow resistance. For example, in some applications it may be the case that a very large reduction in droplet thickness outweighs negative effects of the increased droplet area. In other examples, a small reduction in droplet thickness may be sufficient, such as in cases where a further reduction would not reduce temperature change time sufficiently to speed the overall process or where increased flow resistance should be avoided. A reduction of about 50% is contemplated to be a suitable balance for many applications, including various PCR processes.
  • an electrode 118 that is aligned with the narrowed portion 110 of the gap 106 is energized to attract a droplet that is located at a position 120 outside the narrowed portion 110.
  • an electrode 116 where the droplet is located may be deenergized to release the droplet.
  • the droplet may move from a normal position 120 to a thinning position 122 inside the narrowed portion 110 of the gap 106.
  • the thermal element 114 is then activated to apply a thermal bias to the droplet at the thinning position 122.
  • the thermal element 114 may be continuously active.
  • the electrode 118 that is aligned with the narrowed portion 1 10 may be deenergized to release the droplet and another electrode may be energized to draw the droplet away from its position 122 within the narrowed portion 110 of the gap 106.
  • an array of electrodes 1 12 may include a first electrode 116 at a first site to draw a fluid droplet to the first site that is coincident with the first electrode 116.
  • the array of electrodes 112 may further include a second electrode 118 coincident with a second site to draw the fluid droplet to the second site.
  • the second site may be located at a narrowed portion 110 of a gap between bodies, such as a substrate and cover, a thermal element is positioned.
  • the first and second electrodes 116, 1 18 may be selectively charged and discharged to move the droplet from the first site, where it has original thickness and does not undergo a temperature change, to the second site, where its thickness is reduced and thermal bias is applied.
  • the second site may be about equal in area to the first site.
  • the first and second electrodes 116, 118 may be rectangular and have lengths L and widths W that provide the same area. In various examples, most or all of the electrodes of the array 112 have the same length and width and thus the same area.
  • a device 300 includes multiple contiguous sites 302 where thermal bias may be applied as well as other sites 304, where thermal bias is not applied. Separate electrodes may be provided for different thermal biasing sites 302 or an electrode maybe shared among thermal biasing sites 304. It is noted that since electrodes define sites, these terms may be used interchangeably herein.
  • a narrowed portion 306 of a gap between bodies that define the sites 302, 304 may extend throughout the contiguous temperature change sites 302.
  • a thermal element 308 may extend through the contiguous temperature change sites 302. Separate thermal elements 308 may be provided for different temperature change sites 302 or a thermal element 308 may be shared among temperature change sites 302, as depicted.
  • a temperature change site 302 may be about equal in area to a nontemperature change site 304. Accordingly, the multiple temperature change sites 302 may accommodate the increased area of a droplet 312 due to its reduced thickness at the temperature change sites 302, as compared to a normal thickness droplet 314 at a non-temperature change site 304. Any number of temperature change sites 302 may be provided to accommodate a droplet 312 enlarged by any factor, such as 2, 3, 4, etc. due to reduced thickness.
  • the fluid volume of a non-temperature change site 304 and the total fluid volume of the temperature change sites 302 may be approximately equal. This may allow volumetric flow rate of fluid droplets to be maintained between the non-temperature change site 304 and the temperature change sites 302.
  • Separate thermal elements 308 of the temperature change sites 302 may be controlled in unison or in a synchronized manner to apply a thermal bias a thinned but enlarged droplet 302.
  • a temperature change site 402 at a location between bodies with a reduced gap size 406 may be larger in area than a nontemperature change site 304 with normal or non-reduced gap size.
  • a thermal element 408 located at the temperature change site 402 may be sized consistently with the temperature change site 402.
  • a temperature change site 402 and a non-temperature change site 304 may have areas selected to define fluid volumes that are approximately equal, so that the volumetric flowrate of droplets between the temperature change site 402 and the non-temperature change site 304 may be maintained.
  • the temperature change site 402 has about four times the area as each non-temperature change site 304 and thus has one quarter the gap height, so as to provide the approximately the same volume.
  • FIG. 5 shows an example device 500 for use with a nucleic acid process, such as a PGR process.
  • the device 500 may be referred to as a digital microfluidics device.
  • the device 500 includes an array of electrodes 502 that define an array of sites. Each site may have its electrode 502 selectively charged to draw fluid to the site 502.
  • the charging and discharging of the electrodes 502 at the sites may be programmable, so that an arbitrary path or circuit through select sites may be configured for droplet flow at any given time.
  • the device 500 further includes a plurality of heaters 504, 506, 508 positioned to establish a plurality of temperature zones.
  • the temperature zones may be coincident with regions 604, 606, 608 of reduced gap thickness, as shown in FIG. 6, where the gap between upper and lower bodies 610, 612 is locally reduced compared to a normal or wider gap 614.
  • the gap distance at regions 604, 606, 608 may be the same or different.
  • the device 500 further includes chambers 510, 512, 514, 516 to store components of the nucleic acid process.
  • a chamber 510 stores a master mix
  • a chamber 512 stored eluted DNA with master mix
  • a chamber 514 stores magnetic beads
  • a chamber 516 that stores an elution buffer. Fluid may be selectively moved into and out of the chambers 510, 512, 514, 516 through operation of the electrodes 502 at communicating sites.
  • the chambers 510, 512, 514, 516 may be provided with inlets to receive fluid or other material via an external device, such as a pipet.
  • the device 500 may be controlled to form a circuit 520 (shaded squares) through which droplets are cycled. Electrodes 502 at the sites may be controlled to move droplets to and from the influence of the heaters 504, 506, 508, which may control the droplets to reach respective temperatures of 95 C, 75 C, and 60 C, for example, to effect PCR temperature cycling.
  • a droplet 620 may occupy a region of normal gap 614 due to operation of a corresponding electrode 630.
  • the droplet 620 may be under transport between temperature change sites defined by the heaters 506, 508.
  • Another droplet 622 may occupy a region of narrowed gap 606 due to operation of corresponding electrodes 630, which bring the droplet 622 under the influence of the heater 506.
  • the droplet 622 may undergo heating and its thinned form may reduce the time it takes for the droplet to reach the target temperature (e.g., 75 C).
  • Another droplet 624 may occupy a region of normal gap 614 due to operation of a corresponding electrode 634.
  • the droplet 624 may be under transport between temperature change sites defined by the heaters 504, 506.
  • the droplets 620, 622, 624 depicted represent the movement of a single droplet.
  • An imaging device 640 such as a photosensor or camera, may be provided to capture light emitted by fluorescence of a droplet 624 for target nucleic acid quantification in the PCR process.
  • the upper body 610 may be transparent or semi-transparent to allow for transmission of light for capture by the imaging device 640.
  • the imaging device 640 may be positioned at a region of normal or wider gap 614, so that the measured droplet 624 provides a sufficient or expected degree of fluorescence. That is, because the droplet 624 is not thinned, a sufficient or expected amount of light can be captured by the imaging device 640.
  • the device 500 may further include a controller 530, a non-transitory machine-readable medium 532, and a set of instructions 534.
  • the controller 530 may be connected to the medium 532, the electrodes 502, the heaters 504, 506, 508, and the imaging device 640.
  • the controller 530 may include a processor, a microcontroller, a microprocessor, a processing core, a field-programmable gate array (FPGA), an application-specific integrated circuit (ASIC), or a similar device capable of executing instructions.
  • the processor 530 may cooperate with the non-transitory machine-readable medium 532, which may include an electronic, magnetic, optical, or other physical storage device that encodes the instructions 534.
  • the machine-readable medium 532 may include, for example, random access memory (RAM), read-only memory (ROM), electrically-erasable programmable read-only memory (EEPROM), flash memory, a storage drive, an optical device, or similar.
  • the instructions 534 may be directly executed, such as a binary file, and/or may include interpretable code, bytecode, source code, or similar instructions that may undergo additional processing to be executed. All of such examples may be considered executable instructions.
  • the instructions 534 may control the electrodes 502 to move droplets among the various sites and to cause the droplets to thin in the effective range of the heaters 504, 506, 508, so as to implement the PCR process.
  • the instructions 534 may control the heaters 504, 506, 508 and may further control the imaging device 640.
  • any of the heaters 504, 506, 508 may be replaced by cooling elements to implement a process that uses cooling.
  • thermal cycling in a PGR process may include cooling of droplets between applications of heat.
  • An example method includes controlling an electrode array to flow a fluid droplet from a first site between a pair of opposing bodies to a second site between the pair of opposing bodies. A second distance between the opposing bodies at the second site is smaller than a first distance between the opposing bodies at the first site. The example method further includes applying a thermal bias to the fluid droplet at the second site.
  • the method may further include controlling the electrode array to flow the fluid droplet from the second site to a third site between the pair of opposing bodies.
  • a third distance between the opposing bodies at the third site may be greater than the second distance.
  • the method may further include maintaining a constant average volumetric flow rate of fluid droplets through the first, second, and third sites.
  • the method may further include controlling the electrode array to flow the fluid droplet from the third site to a fourth site between the pair of opposing bodies.
  • a fourth distance between the opposing bodies at the fourth site may be smaller than the third distance.
  • the method may further include applying a thermal bias to the fluid droplet at the fourth site to a obtain a temperature that may be different from the temperature of the second site. The fourth distance may be different from the second distance.
  • the method may further include performing nucleic acid denaturation, annealing, or elongation of a polymerase chain reaction with the fluid droplet at the second site.
  • FIG. 7 shows an example method 700 of transporting a droplet to a narrowed site between bodies and applying a thermal bias to a droplet at the site.
  • the method 700 may be implemented by executable instructions, such as instructions 534 discussed above.
  • an electrode array is controlled to flow a fluid droplet 710 from a first site 706 between a pair of opposing bodies to a second site 708 between the pair of opposing bodies.
  • An electrode at the first site 706 may be deenergized and, at about the same time, an electrode at the second site 708 may be energized.
  • the second site 708 thins the fluid droplet 710, so that a temperature change may be effected more quickly than at the first site 706.
  • a second distance D2 between the opposing bodies at the second site 708 is smaller than a first distance D1 between the opposing bodies at the first site 706.
  • a thermal bias Q is applied to the fluid droplet 710 at the second site 708.
  • a heater such as a resistive heating element or heat pipe, may be used to heat the droplet 710.
  • a cooling device such as a thermoelectric device or heat pipe, may be used to cool the droplet 710. Heating or cooling may be active or passive.
  • Heating at the second site 708 may be part of thermal cycling in a PCR process.
  • the heating may be applied to achieve temperature sufficient for nucleic acid denaturation, annealing, or elongation.
  • FIG. 8 shows an example method 800 of transporting a droplet to sites that provide various thickness to the droplet and applying a thermal bias to a droplet at a narrowed site.
  • the method 800 may be implemented by executable instructions, such as instructions 534 discussed above.
  • Reference to the method 700 may be made for details not repeated here, where like reference numerals and like terminology denote like elements.
  • an electrode array is controlled to flow a fluid droplet 710 from a first site 706 between a pair of opposing bodies to a second site 708 between the pair of opposing bodies.
  • a thermal bias Q1 is applied to the fluid droplet 710 at the second site 708.
  • the electrode array is controlled to flow the fluid droplet 710 from the second site 708 to a third site 808 that allows the droplet 710 to thicken.
  • a third distance D3 of the gap at the third site 808 is greater than the second distance D2 at the second site.
  • the third distance D3 may be the same as or different from the first distance D1 at the first site 706.
  • Application of thermal bias may be avoided at the third site 808.
  • a constant average volumetric flow rate of fluid droplets may be maintained through the first, second, and third sites 706, 708, 808.
  • the electrode array is controlled to flow the droplet 710 from the third site 808 to a fourth site 810 that thins the droplet.
  • a fourth distance D4 between the opposing bodies at the fourth site 810 is smaller than the third distance D3 at the third site 808.
  • the fourth distance D4 may be the same as or different from the second distance D2 at the second site 708.
  • a thermal bias Q2 is applied to the droplet 710 at the fourth site 810 to obtain a temperature that may be different from the temperature of the droplet 710 reached at the second site 708.
  • the distances D1 , D2, D3, D4 and thermal biases Q1 , Q2 may be selected according to the specific chemical, biological, or biochemical process being implemented.
  • the method 800 may circulate the droplet 710 by conveying the droplet 710 from the fourth site 810 to the first site 706.
  • the method 800 may include additional sites of varying gap distance and thermal input.
  • a droplets thickness this reduced by half from D to D/2, it may undergo a temperature change twice as fast, all other things being equal. That, is a time tD/2 for a thinned droplet to reach a target temperature T may be half a time to for a non-thinned droplet to reach the same target temperature T.
  • a locally narrowed gap in a microfluidic device allows for thinning of a droplet during heating or cooling and therefore a higher rate of heating or cooling.
  • Thermocycling times may be reduced, as may overall assay time. Reduction of assay time allows for more efficient nucleic acid testing and faster decision making.
  • the thinning of the droplet is localized and the droplet may be thicker at locations where thickness has utility, such as where fluorescence measurements are made.

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Abstract

Un exemple de dispositif comprend un substrat et un couvercle séparé du substrat par un espace qui comprend une partie rétrécie. L'espace restreint une épaisseur d'une gouttelette de fluide située entre le substrat et le couvercle. Le dispositif comprend en outre un réseau d'électrodes au niveau du substrat. Le réseau d'électrodes déplace la gouttelette de fluide à l'intérieur de l'espace en réponse à une charge appliquée à une électrode du réseau d'électrodes. Le dispositif comprend en outre un élément thermique positionné par rapport au substrat pour modifier une température de la gouttelette de fluide. L'élément thermique est positionné au niveau de la partie rétrécie de l'espace pour réduire l'épaisseur de la gouttelette de fluide tout en subissant le changement de température.
PCT/US2020/057181 2020-10-23 2020-10-23 Microfluidique ayant des épaisseurs de gouttelettes réduites Ceased WO2022086565A1 (fr)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130116128A1 (en) * 2011-11-07 2013-05-09 Illumina, Inc. Integrated sequencing apparatuses and methods of use
US20160199832A1 (en) * 2013-08-30 2016-07-14 Advanced Liquid Logic France Sas Manipulation of droplets on hydrophilic or variegated-hydrophilic surfaces

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130116128A1 (en) * 2011-11-07 2013-05-09 Illumina, Inc. Integrated sequencing apparatuses and methods of use
US20160199832A1 (en) * 2013-08-30 2016-07-14 Advanced Liquid Logic France Sas Manipulation of droplets on hydrophilic or variegated-hydrophilic surfaces

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