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WO2025106391A1 - Ensemble puce microfluidique avec train roulant métallique - Google Patents

Ensemble puce microfluidique avec train roulant métallique Download PDF

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Publication number
WO2025106391A1
WO2025106391A1 PCT/US2024/055438 US2024055438W WO2025106391A1 WO 2025106391 A1 WO2025106391 A1 WO 2025106391A1 US 2024055438 W US2024055438 W US 2024055438W WO 2025106391 A1 WO2025106391 A1 WO 2025106391A1
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WIPO (PCT)
Prior art keywords
chamber portion
fluid
layer
recess
sidewall
Prior art date
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English (en)
Inventor
Babak Sanii
James Johnson
Ximiao WEN
Ping-wei CHEN
Ning Yu
Wei Cai
Casey FUOCO
Benjamin Eldridge
Jason COSMAN
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Nutcracker Therapeutics Inc
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Nutcracker Therapeutics Inc
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Publication of WO2025106391A1 publication Critical patent/WO2025106391A1/fr
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    • 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
    • B01L3/50273—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 the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • 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
    • B01L3/502707—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 the manufacture of the container or its components
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    • 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
    • B01L3/502738—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 integrated valves
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
    • F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
    • F04B19/00—Machines or pumps having pertinent characteristics not provided for in, or of interest apart from, groups F04B1/00 - F04B17/00
    • F04B19/006—Micropumps
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
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    • B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
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    • B01L2300/08—Geometry, shape and general structure
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    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • 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
    • B01L2400/0481—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
    • 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
    • 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
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
    • 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
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0633—Valves, specific forms thereof with moving parts
    • B01L2400/0655—Valves, specific forms thereof with moving parts pinch valves
    • 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

Definitions

  • polynucleotide therapeutics e.g., mRNA therapeutics, etc.
  • Some currently available technologies for manufacturing and formulating polynucleotide therapeutics may expose the products to contamination and degradation.
  • Some available centralized production may be too costly, too slow, or susceptible to contamination for use in therapeutic formulations possibly including multiple polynucleotide species.
  • FIG. 1 depicts a schematic view of an example of a system including a microfluidic process chip
  • FIG. 2 depicts an exploded perspective view of examples of components of the system of FIG. 1;
  • FIG. 3 depicts a top plan view of an example of a process chip that may be incorporated into the system of FIG. 1;
  • FIG. 4 A depicts a cross-sectional side view of the process chip of FIG. 3 in a first state of operation
  • FIG. 4B depicts a cross-sectional side view of the process chip of FIG. 3 in a second state of operation
  • FIG. 4C depicts a cross-sectional side view of the process chip of FIG. 3 in a third state of operation
  • FIG. 4D depicts a cross-sectional side view of the process chip of FIG. 3 in a fourth state of operation
  • FIG. 4E depicts a cross-sectional side view of the process chip of FIG. 3 in a fifth state of operation
  • FIG. 4F depicts a cross-sectional side view of the process chip of FIG. 3 in a sixth state of operation
  • FIG. 5 depicts a perspective view of another example of a process chip that may be incorporated into the system of FIG. 1;
  • FIG. 6 depicts another perspective view of the process chip of FIG. 5;
  • FIG. 7 depicts a top plan view of the process chip of FIG. 7, with certain layers being transparent to show internal features;
  • FIG. 8 depicts an exploded perspective view of the process chip of FIG. 5;
  • FIG. 9 depicts a perspective view of a first layer of the process chip of FIG. 5;
  • FIG. 10 depicts another perspective view of the first layer of FIG. 9;
  • FIG. 11 depicts a bottom plan view of the first layer of FIG. 9;
  • FIG. 12 depicts a perspective view of a third layer of the process chip of FIG. 5;
  • FIG. 13 depicts another perspective view of the third layer of FIG. 12;
  • FIG. 14 depicts a top plan view of the third layer of FIG. 12;
  • FIG. 15 depicts a perspective view of a fourth layer of the process chip of FIG. 5;
  • FIG. 16 depicts a perspective view of a sixth layer of the process chip of FIG. 5;
  • FIG. 17 depicts another perspective view of the sixth layer of FIG. 16;
  • FIG. 18 depicts a top plan view of the sixth layer of FIG. 16;
  • FIG. 19 depicts a perspective view of a combination of the third layer of FIG. 12, the fourth layer of FIG. 15, a fifth layer of the process chip of FIG. 5, and the sixth layer of FIG. 16;
  • FIG. 20 depicts a top plan view of the combination of layers of FIG. 19;
  • FIG. 21 depicts an enlarged perspective view of a portion of the combination of layers of FIG. 19;
  • FIG. 22A depicts a cross-sectional side view, taken along line 22-22 of FIG. 21, of a portion of the process chip of FIG. 5, with a second layer of the process chip in a non-deformed state;
  • FIG. 22B depicts a cross-sectional side view, taken along line 22-22 of FIG. 21, of the portion of FIG. 22 A, with the second layer of the process chip in a deformed state;
  • FIG. 23 A depicts a cross-sectional side view, taken along line 23-23 of FIG. 21, of another portion of the process chip of FIG. 5, with the second layer of the process chip in a non-deformed state;
  • FIG. 23B depicts a cross-sectional side view of the portion of FIG. 23 A, taken along line 23-23 of FIG. 21, with the second layer of the process chip in a deformed state;
  • FIG. 24 depicts an enlarged perspective view of another portion of the combination of layers of FIG. 19;
  • FIG. 25 depicts an enlarged perspective view of a fluid inlet region of the portion of FIG. 24;
  • FIG. 26 depicts an enlarged perspective view of a fluid outlet region of the portion of FIG. 24;
  • FIG. 27 depicts a perspective view of another example of a process chip that may be incorporated into the system of FIG. 1;
  • FIG. 28 depicts another perspective view of the process chip of FIG. 27;
  • FIG. 29 depicts a top plan view of the process chip of FIG. 27, with certain layers being transparent to show internal features;
  • FIG. 30 depicts an exploded perspective view of the process chip of FIG. 27;
  • FIG. 31 depicts a perspective view of a third layer of the process chip of FIG. 27;
  • FIG. 32 depicts another perspective view of the third layer of FIG. 31;
  • FIG. 33 depicts a bottom plan view of the third layer of FIG. 31;
  • FIG. 34 depicts a perspective view of a fourth layer of the process chip of FIG. 27;
  • FIG. 35 depicts a perspective view of a fifth layer of the process chip of FIG. 27;
  • FIG. 36 depicts a perspective view of a sixth layer of the process chip of FIG. 27;
  • FIG. 37 depicts a top plan view of the sixth layer of FIG. 36;
  • FIG. 38 depicts a perspective view of a combination of the third layer of FIG.
  • FIG. 39 depicts an enlarged perspective view of a portion of the combination of layers of FIG. 38.
  • FIG. 40 depicts a cross-sectional side view of a portion of the combination of layers of FIG. 38, taken along line 40-40 of FIG. 39.
  • apparatuses and methods are disclosed herein for processing therapeutic polynucleotides.
  • these apparatuses and methods may be closed path apparatuses and methods that are configured to minimize or eliminate manual handling during operation.
  • the closed path apparatuses and methods may provide a nearly entirely aseptic environment, and the components may provide a sterile path for processing from initial input (e.g., template) to output (e.g., compounded therapeutic).
  • Material inputs e.g., nucleotides, and any chemical components
  • into the apparatus may be sterile; and may be input into the system without requiring virtually any manual interaction.
  • the apparatuses and methods described herein may be used to generate therapeutics at rapid cycle times at high degree of reproducibility.
  • the apparatuses described herein may be configured to provide, in a single integrated apparatus, synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions. Alternatively, one or more of these processes may be carried out in two or more apparatuses as described herein.
  • the therapeutic compositions may include therapeutic polynucleotides, such as, for example, ribonucleic acids or deoxyribonucleic acids.
  • the polynucleotides may include only natural nucleotide units or may include any kind of synthetic, semi-synthetic, or modified nucleotide units.
  • All or some of the processing steps may be performed in an unbroken fluid processing pathway, which may be configured as one or a series of consumable microfluidic path device(s) — in some instances also referred to herein as a process chip or a biochip (though the chip need not necessarily be used in bio-related applications).
  • the process chip in some examples may be removably installed in an instrument that is part of a larger microfluidic system, such as that shown in FIG. 1.
  • the disclosed apparatuses and methods may be used for the synthesis of patient-specific therapeutics, including compounding, at a point of care (e.g., hospital, clinic, pharmacy, etc.).
  • any of the apparatuses and methods described herein should be understood to be inclusive, but all or a sub-set of the components and/or steps may alternatively be exclusive and may be expressed as “consisting of’ or alternatively “consisting essentially of’ the various components, steps, sub-components, or sub-steps.
  • spatially relative terms such as “under,” “below,” “lower,” “over,” “upper,” and the like, may be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is inverted, elements described as “under” or “beneath” other elements or features would then be oriented “over” the other elements or features. Thus, the term “under” may encompass both an orientation of over and under.
  • the device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.
  • the terms “upwardly,” “downwardly,” “vertical,” “horizontal,” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.
  • a feature or element When a feature or element is herein referred to as being “on” another feature or element, it may be directly on the other feature or element or intervening features and/or elements may also be present. In contrast, when a feature or element is referred to as being “directly on” another feature or element, there are no intervening features or elements present.
  • a feature or element When a feature or element is referred to as being “connected,” “attached,” or “coupled” to another feature or element, it may be directly connected, attached, or coupled to the other feature or element or intervening features or elements may be present. In contrast, when a feature or element is referred to as being “directly connected,” “directly attached,” or “directly coupled” to another feature or element, there are no intervening features or elements present.
  • references to a structure or feature that is disposed “adjacent” another feature may have portions that overlap or underlie the adjacent feature.
  • a numeric value may have a value that is ⁇ 0.1% of the stated value (or range of values), ⁇ 1% of the stated value (or range of values), ⁇ 2% of the stated value (or range of values), ⁇ 5% of the stated value (or range of values), ⁇ 10% of the stated value (or range of values), etc.
  • Any numerical values given herein should also be understood to include about or approximately that value unless the context indicates otherwise. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Any numerical range recited herein is intended to include all sub-ranges subsumed therein.
  • first and second may be used herein to describe various features/elements (including steps), these features/elements should not be limited by these terms, unless the context indicates otherwise. These terms are used to distinguish one feature/element from another feature/element, and unless specifically pointed out, do not denote a certain order. Thus, a first feature/element discussed below could be termed a second feature/element, and similarly, a second feature/element discussed below could be termed a first feature/element without departing from the teachings of the present invention.
  • system As used herein, the terms “system,” “apparatus,” and “device” may be read as being interchangeable with each other.
  • a system, apparatus, and device may each include a plurality of components having various kinds of structural and/or functional relationships with each other.
  • polynucleotide refers to a nucleic acid molecule containing multiple nucleotides and generally refers both to “oligonucleotides” (a polynucleotide molecule of 18-25 nucleotides in length) and polynucleotides of 26 or more nucleotides.
  • compositions including oligonucleotides having a length of 18-25 nucleotides (e.g., 18-mers, 19-mers, 20-mers, 21-mers, 22-mers, 23- mers, 24-mers, or 25-mers), or medium-length polynucleotides having a length of 26 or more nucleotides (e.g., polynucleotides of 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 210, about 220, about 230, about 240, about 250, about 260, about
  • amplification may refer to polynucleotide amplification.
  • Amplification may include any suitable method for amplification of a polynucleotide and includes, but is not limited to, multiple displacement amplification (MDA), polymerase chain reaction (PCR) amplification, Loop Mediated Isothermal Amplification (LAMP), Nucleic Acid Sequence Based Amplification, Strand Displacement Amplification, Rolling Circle Amplification, and Ligase Chain Reaction.
  • MDA multiple displacement amplification
  • PCR polymerase chain reaction
  • LAMP Loop Mediated Isothermal Amplification
  • Nucleic Acid Sequence Based Amplification Strand Displacement Amplification
  • Rolling Circle Amplification Rolling Circle Amplification
  • Ligase Chain Reaction Ligase Chain Reaction
  • a “cassette” refers to a polynucleotide sequence which may include or be operably linked to one or more expression elements such as an enhancer, a promoter, a leader, an intron, a 5' untranslated region (UTR), a 3' UTR, or a transcription termination sequence.
  • a cassette comprises at least a first polynucleotide sequence capable of initiating transcription of an operably linked second polynucleotide sequence (which may comprise a template) and optionally a transcription termination sequence operably linked to the second polynucleotide sequence.
  • the template as described below, may comprise a sequence of interest, for example, an open reading frame (“ORF”) of interest.
  • the cassette may be provided as a single element or as two or more unlinked elements.
  • a “template” refers to a nucleic acid sequence that contains a sequence of interest for preparing a therapeutic polynucleotide according to the disclosed methods. Templates may be, but are not limited to, a double stranded DNA (dsDNA), an engineered plasmid construct, a cDNA sequence, or a linear nucleic acid sequence (for example, a linear template generated by PCR or by annealing chemically synthesized oligonucleotides). The template may, in certain aspects, be integrated into a “cassette” as described above.
  • sequence of interest refers to a polynucleotide sequence, the use of which may be deemed desirable for a suitable purpose, in particular, for the manufacture of an mRNA for a therapeutic use, and includes but is not limited to, coding sequences of structural genes, and non-coding regulatory sequences that do not encode and mRNA or protein product.
  • zw vitro transcription or “IVT” refer to the process whereby transcription occurs in vitro in a non-cellular system to produce synthetic RNA molecules (e.g., synthetic mRNA) for use in various applications, including for therapeutic delivery to a subject, for example, as a therapeutic polynucleotide, which may be part of, or may be used to form, a therapeutic polynucleotide composition as described below.
  • the therapeutic polynucleotide, (e.g., synthetic RNA molecules (transcription product)) generated may be combined with a delivery vehicle to form a therapeutic polynucleotide composition.
  • Synthetic transcription products include mRNAs, antisense RNA molecules, shRNA, circular RNA molecules, ribozymes, and the like.
  • An IVT reaction may use a purified linear DNA template comprising a promoter sequence and the sequence of the open reading frame (ORF) of a sequence of interest, ribonucleotide triphosphates or modified ribonucleotide triphosphates, a buffer system that includes DTT and magnesium ions, and a phage RNA polymerase.
  • ORF open reading frame
  • a “therapeutic polynucleotide” refers to a polynucleotide (e.g., an mRNA) that may be part of a therapeutic polynucleotide composition for delivery to a subject to treat a symptom, disease, or condition in a subject; prevent a symptom, disease, or condition in a subject; or to improve or otherwise modify the subject’s health.
  • a polynucleotide e.g., an mRNA
  • a “therapeutic polynucleotide composition” may refer to a composition including one or more therapeutic polynucleotides (e.g., mRNA) encapsulated by a delivery vehicle, which composition may be administered to a subject in need thereof using any suitable administration routes, such as intratumoral, intramuscular, etc. injection.
  • a therapeutic polynucleotide composition is an mRNA (therapeutic) nanoparticle comprising at least one mRNA encapsulated by a delivery vehicle molecule.
  • An mRNA vaccine is one example of a therapeutic polynucleotide composition.
  • delivery vehicle refers to any substance that facilitates, at least in part, the in vivo, in vitro, or ex vivo delivery of a polynucleotide (e.g., therapeutic polynucleotide) to targeted cells or tissues (e.g., tumors, etc.).
  • a delivery vehicle need not exclude the possibility of the delivery vehicle also having therapeutic effects.
  • Some versions of a delivery vehicle may provide additional therapeutic effects.
  • a delivery vehicle may be a peptoid molecule, such as an amino-lipidated peptoid molecule, that may be used to at least partially encapsulate mRNA.
  • the term “DV” will also be used herein as a shorthand for “delivery vehicle.”
  • joining refers to methods such as ligation, synthesis, primer extension, annealing, recombination, or hybridization use to couple one component to another.
  • purifying refers to physical and/or chemical separation of a component (e.g., particles) of other unwanted components (e.g., contaminating substances, fragments, etc.).
  • the term “substantially free” as used with respect to a given substance includes 100% free of a given substance, or which comprises less than about 1.0%, or less than about 0.5%, or less than about 0.1% of the given substance.
  • FIG. 1 depicts examples of various components that may be incorporated into a system (100).
  • System (100) of this example includes a housing (103) enclosing a seating mount (115) that may removably hold one or more microfluidic process chips (111).
  • system (100) includes a chip-receiving component that is configured to removably accommodate a process chip (111), where the process chip (111) itself defines one or more microfluidic channels or fluid pathways.
  • Components of system (100) may include fluid channels or pathways that are not necessarily considered microfluidic (e.g., with such fluid channels or pathways being larger than the microfluidic channels or fluid pathways in process chip (111)).
  • process chips (111) are provided and utilized as single-use devices, while the rest of system (100) is reusable.
  • Housing (103) may be in the form of a chamber, enclosure, etc., with an opening that may be closed (e.g., via a lid or door, etc.) to thereby seal the interior.
  • Housing (103) may enclose a thermal regulator and/or may be configured to be enclosed in a thermally-regulated environment (e.g., a refrigeration unit, etc.). Housing (103) may form an aseptic barrier. In some variations, housing (103) may form a humidified or humidity-controlled environment. In addition, or in the alternative, system (100) may be positioned in a cabinet (not shown). Such a cabinet may provide a temperature-regulated (e.g., refrigerated) environment. Such a cabinet may also provide air filtering and air flow management and may promote reagents being kept at a desired temperature through the manufacturing process. In addition, such a cabinet may be equipped with UV lamps for sterilization of process chip (111) and other components of system (100). Other suitable features may be incorporated into a cabinet that houses system (100).
  • a thermally-regulated environment e.g., a refrigeration unit, etc.
  • Housing (103) may form an aseptic barrier. In some variations, housing (103) may form a humidified or humidity-controlled environment.
  • the assembly formed by housing (103) and the components of system (100) that are within housing (103), without process chip (111), may be considered as being an “instrument.” While controller (121) and user interface (123) are shown in FIG. 1 as being outside of housing (103), controller (121) and user interface (123) may in fact be provided in or on housing (103) and may thus also form part of the instrument. As described in greater detail below, this instrument may removably receive process chip (111) via a seating mount (115). When process chip (111) is seated in seating mount (115), the instrument and process chip (111) cooperate to together form system (100).
  • process chip (111) When process chip (111) is removed from seating mount (115), the portion of system (100) that is left may be regarded as the “instrument.”
  • the instrument, the system (100), and process chip (111) may each be considered an “apparatus.”
  • the term “apparatus” may thus be read to include the instrument by itself, a process chip (111) by itself, the combination of the instrument and process chip (111), some other combination of components of system (100), or some other permutation of system (100) or components thereof.
  • Seating mount (115) may be configured to secure process chip (111) using one or more pins or other components configured to hold process chip (111) in a fixed and predefined orientation. Seating mount (115) may thus facilitate process chip (111) being held at an appropriate position and orientation in relation to other components of system (100). In the present example, seating mount (115) is configured to hold process chip (111) in a horizontal orientation, such that process chip (111) is parallel with the ground.
  • a thermal control (113) may be located adjacent to seating mount (115), to modulate the temperature of any process chip (111) mounted in seating mount (115).
  • Thermal control (113) may include a thermoelectric component (e.g., Peltier device, etc.) and/or one or more heat sinks for controlling the temperature of all or a portion of any process chip (111) mounted in seating mount (115).
  • more than one thermal control (113) may be included, such as to separately regulate the temperature of different ones of one or more regions of process chip (111).
  • Thermal control (113) may include one or more thermal sensors (e.g., thermocouples, etc.) that may be used for feedback control of process chip (111) and/or thermal control (H3).
  • a fluid interface assembly (109) couples process chip (111) with a pressure source (117), thereby providing one or more paths for fluid (e.g., gas) at a positive or negative pressure to be communicated from pressure source (117) to one or more interior regions of process chip (111) as will be described in greater detail below.
  • system (100) may include two or more pressure sources (117).
  • pressure may be generated by one or more sources other than pressure source (117).
  • one or more vials or other fluid sources within reagent storage frame (107) may be pressurized.
  • reactions and/or other processes carried out on process chip (111) may generate additional fluid pressure.
  • fluid interface assembly (109) also couples process chip (111) with a reagent storage frame (107), thereby providing one or more paths for liquid reagents, etc., to be communicated from reagent storage frame (107) to one or more interior regions of process chip (111) as will be described in greater detail below.
  • pressurized fluid e.g., gas
  • reagent storage frame (107) includes one or more components interposed in the fluid path between pressure source (117) and fluid interface assembly (109).
  • one or more pressure sources (117) are directly coupled with fluid interface assembly, such that the positively pressurized fluid (e.g., positively pressurized gas) or negatively pressurized fluid (e.g., suction or other negatively pressurized gas) bypasses reagent storage frame (107) to reach fluid interface assembly (109).
  • fluid interface assembly (109) may be removably coupled to the rest of system (100), such that at least a portion of fluid interface assembly (109) may be removed for sterilization between uses.
  • pressure source (117) may selectively pressurize one or more chamber regions on process chip (111).
  • pressure source may also selectively pressurize one or more vials or other fluid storage containers held by reagent storage frame (107).
  • Reagent storage frame (107) is configured to contain a plurality of fluid sample holders, each of which may hold a fluid vial that is configured to hold a reagent (e.g., nucleotides, solvent, water, etc.) for delivery to process chip (111).
  • a fluid vial that is configured to hold a reagent (e.g., nucleotides, solvent, water, etc.) for delivery to process chip (111).
  • a reagent e.g., nucleotides, solvent, water, etc.
  • one or more fluid vials or other storage containers in reagent storage frame (107) may be configured to receive a product from the interior of the process chip (111).
  • a second process chip (111) may receive a product from the interior of a first process chip (111), such that one or more fluids are transferred from one process chip (111) to another process chip (111).
  • the first process chip (111) may perform a first dedicated function (e.g., synthesis, etc.) while the second process chip (111) performs a second dedicated function (e.g., encapsulation, etc.).
  • Reagent storage frame (107) of the present example includes a plurality of pressure lines and/or a manifold configured to divide one or more pressure sources (117) into a plurality of pressure lines that may be applied to process chip (111). Such pressure lines may be independently or collectively (in subcombinations) controlled.
  • Fluid interface assembly (109) may include a plurality of fluid lines and/or pressure lines where each such line includes a biased (e.g., spring-loaded) holder or tip that individually and independently drives each fluid and/or pressure line to process chip (111) when process chip (111) is held in seating mount (115).
  • Any associated tubing e.g., the fluid lines and/or the pressure lines
  • each fluid line comprises a flexible tubing that connects between reagent storage frame (107), via a connector that couples the vial to the tubing in a locking engagement (e.g., ferrule) and process chip (111).
  • the ends of the fluid lines/pressure lines may be configured to seal against process chip (111) (e.g., at a corresponding sealing port formed in process chip (111)), as described below.
  • process chip (111) e.g., at a corresponding sealing port formed in process chip (111)
  • the connections between pressure source (117) and process chip (111), and the connections between vials in reagent storage frame (107) and process chip (111) all form sealed and closed paths that are isolated when process chip (111) is seated in seating mount (115). Such sealed, closed paths may provide protection against contamination when processing therapeutic polynucleotides.
  • the vials of reagent storage frame (107) may be pressurized (e.g., > 1 atm pressure, such as 2 atm, 3 atm, 5 atm, or higher).
  • the vials may be pressurized by pressure source (117). Negative or positive pressure may thus be applied.
  • the fluid vials may be pressurized to between about 1 and about 20 psig (e.g., 5 psig, 10 psig, etc.).
  • a vacuum e.g., about -7 psig or about 7 psia
  • System (100) of the present example further includes a magnetic field applicator (119), which is configured to create a magnetic field at a region of the process chip (111).
  • Magnetic field applicator (119) may include a movable head that is operable to move the magnetic field to thereby selectively isolate products that are adhered to magnetic capture beads within vials or other storage containers in reagent storage frame (107).
  • System (100) of the present example further includes one or more sensors (105).
  • sensors (105) include one or more cameras and/or other kinds of optical sensors.
  • Such sensors (105) may sense one or more of a barcode, a fluid level within a fluid vial held within reagent storage frame (107), fluidic movement within a process chip (111) that is mounted within seating mount (115), and/or other optically detectable conditions.
  • a sensor (105) is used to sense barcodes
  • such barcodes may be included on vials of reagent storage frame (107), such that sensor (105) may be used to identify vials in reagent storage frame (107).
  • a single sensor (105) is positioned and configured to simultaneously view such barcodes on vials in reagent storage frame (107), fluid levels in vials in reagent storage frame (107), fluidic movement within a process chip (111) that is mounted within seating mount (115), and/or other optically detectable conditions.
  • more than one sensor (105) is used to view such conditions.
  • different sensors (105) may be positioned and configured to separately view corresponding optically detectable conditions, such that a sensor (105) may be dedicated to a particular corresponding optically detectable condition.
  • sensors (105) include at least one optical sensor
  • visual/optical markers may be used to estimate yield.
  • fluorescence may be used to detect process yield or residual material by tagging with fluorophores.
  • dynamic light scattering DLS
  • sensor (105) may provide measurements using one or two optical fibers to convey light (e.g., laser light) into process chip (111); and detect an optical signal coming out of process chip (111).
  • sensor (105) optically detects process yield or residual material, etc.
  • sensor (105) may be configured to detect visible light, fluorescent light, an ultraviolet (UV) absorbance signal, an infrared (IR) absorbance signal, and/or any other suitable kind of optical feedback.
  • UV ultraviolet
  • IR infrared
  • sensors (105) include at least one optical sensor that is configured to capture video images
  • sensors (105) may record at least some activity on process chip (111).
  • an entire run for synthesizing and/or processing a material e.g., a therapeutic RNA
  • Processing on process chip (111) may be visually tracked and this video record may be retained for later quality control and/or processing.
  • the video record of the processing may be saved, stored, and/or transmitted for subsequent review and/or analysis.
  • the video may be used as a real-time feedback input that may affect processing using at least visually observable conditions captured in the video.
  • Controller (121) may include one or more processors, one or more memories, and various other suitable electrical components.
  • one or more components of controller (121) e.g., one or more processors, etc.
  • system (100) e.g., contained within housing (103).
  • one or more components of controller (121) e.g., one or more processors, etc.
  • controller (121) may be detachably attached or detachably connected with other components of system (100).
  • at least a portion of controller (121) may be removable.
  • at least a portion of controller (121) may be remote from housing (103) in some versions.
  • controller (121) may include activating pressure source (117) to apply pressure through process chip (111) to drive fluidic movement, among other tasks.
  • Controller (121) may be completely or partially outside of housing (103); or completely or partially inside of housing (103).
  • Controller (121) may be configured to receive user inputs via a user interface (123) of system (100); and provide outputs to users via user interface (123).
  • controller (121) is fully automated to a point where user inputs are not needed.
  • user interface (123) may provide only outputs to users.
  • User interface (123) may include a monitor, a touchscreen, a keyboard, and/or any other suitable features.
  • Controller (121) may coordinate processing, including moving one or more fluid(s) onto and on process chip (111), mixing one or more fluids on process chip (111), adding one or more components to process chip (111), metering fluid in process chip (111), regulating the temperature of process chip (111), applying a magnetic field (e.g., when using magnetic beads), etc.
  • Controller (121) may receive real-time feedback from sensors (105) and execute control algorithms in accordance with such feedback from sensors (105).
  • Such feedback from sensors (105) may include, but need not be limited to, identification of reagents in vials in reagent storage frame (107), detected fluid levels in vials in reagent storage frame (107), detected movement of fluid in process chip (111), fluorescence of fluorophores in fluid in process chip (111), etc.
  • Controller (121) may include software, firmware and/or hardware. Controller (121) may also communicate with a remote server, e.g., to track operation of the apparatus, to re-order materials (e.g., components such as nucleotides, process chips (111), etc.), and/or to download protocols, etc.
  • a remote server e.g., to track operation of the apparatus, to re-order materials (e.g., components such as nucleotides, process chips (111), etc.), and/or to download protocols, etc.
  • FIG. 2 shows examples of certain forms that may be taken by various components of system (100).
  • FIG. 2 shows a reagent storage frame (150), a fluid interface assembly (152), a seating mount (154), a thermal control (156), and a process chip (200).
  • Reagent storage frame (150), fluid interface assembly (152), seating mount (154), thermal control (156), and process chip (200) of this example may be configured and operable just like reagent storage frame (107), fluid interface assembly (109), seating mount (115), thermal control (113), and process chip (111), respectively, described above.
  • a set of rods (182) support reagent storage frame (150) over fluid interface assembly (152).
  • a set of optical sensors (160) are positioned at four respective locations along base (180).
  • Optical sensors (160) may be configured and operable like sensors (105) described above.
  • Optical sensors (160) may include off- the-shelf cameras or any other suitable kinds of optical sensors.
  • Optical sensors (160) are positioned such that fluid vials held within reagent storage frame (150) are within the field of view of one or more of optical sensors (160).
  • process chip (200) is within the field of view of one or more of optical sensors (160).
  • Each optical sensor (160) is movably secured to base (180) via a corresponding rail (184) (e.g., in a gantry arrangement), such that each optical sensor (160) is configured to translate laterally along each corresponding rail (184).
  • a linear actuator (186) is secured to each optical sensor (160) and is thereby operable to drive lateral translation of each optical sensor (160) along the corresponding rail (184).
  • Each actuator (186) may be in the form of a drive belt, a drive chain, a drive cable, or any other suitable kind of structure.
  • Controller (121) may drive operation of actuators (186).
  • Optical sensors (160) may be moved along rails (184) during operation of system (100) in order to facilitate viewing of the appropriate regions of vials in reagent storage frame (150) and/or process chip (200). In some scenarios, optical sensors (160) move in unison along corresponding rails (184). In some other scenarios, optical sensors (160) move independently along corresponding rails (184).
  • optical sensors (160) are shown in FIG. 2 as being mounted to base (180), optical sensors (160) may be positioned elsewhere within system (100), in addition to or as an alternative to being mounted to base (180).
  • some versions of reagent storage frame (107) may include one or more optical sensors (160) positioned and configured to provide an overhead field of view.
  • such optical sensors (160) may be mounted to rails, movable cantilever arms, or other structures that allow such optical sensors (160) to be repositioned during operation of system (100).
  • Optical sensors (160) may be positioned in any other suitable locations.
  • system (100) may also include one or more sources of light (e.g., electroluminescent panels, etc.) to provide illumination that aids in optical sensing by optical sensors (160).
  • one or more mirrors are used to facilitate visualization of components of system (100) by optical sensors (160). Such mirrors may allow optical sensors (160) to view components of system (100) that may not otherwise be within the field of view of sensors (160). Such mirrors may be placed directly adjacent to optical sensors (160). In addition, or in the alternative, such mirrors may be placed adjacent to one or more components of system (100) that are to be viewed by optical sensors (160). [0095] In use of system (100), an operator may select a protocol to run (e.g., from a library of preset protocols), or the user may enter a new protocol (or modify an existing protocol), via user interface (123).
  • a protocol to run e.g., from a library of preset protocols
  • the user may enter a new protocol (or modify an existing protocol), via user interface (123).
  • controller (121) may instruct the operator which kind of process chip (111) to use, what the contents of vials in reagent storage frame (107) should be, and where to place the vials in reagent storage frame (107).
  • the operator may load process chip (111) into seating mount (115); and load the desired reagent vials and export vials into reagent storage frame (107).
  • System (100) may confirm the presence of the desired peripherals, identify process chip (111), and scan identifiers (e.g., barcodes) for each reagent and product vial in reagent storage frame (107), facilitating the vials to match the bill-of-reagents for the selected protocol.
  • controller (121) may execute the protocol. During execution, valves and pumps are actuated to deliver reagents as described in greater detail below, reagents are blended, temperature is controlled, and reactions occur, measurements are made, and products are pumped to destination vials in reagent storage frame (107).
  • FIGS. 3 and 4A-4F depict the example of a process chip (200) in further detail.
  • process chip (200) may be utilized to provide in-vitro synthesis, purification, concentration, formulation, and/or analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions.
  • process chip (200) of this example includes a plurality of fluid ports (220). Each fluid port (220) has an associated fluid channel (222) formed in process chip (200), such that fluid communicated into fluid port (220) will flow through the corresponding fluid channel (222).
  • each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109).
  • each fluid channel (222) leads to a valve chamber (224), which is operable to selectively prevent or permit fluid from the corresponding fluid channel (222) to be further communicated along process chip (200) as will be described in greater detail below.
  • process chip (200) of this example includes a plurality of additional chambers (230, 250, 270) that may be used to serve different purposes during the process of producing the therapeutic composition as described herein.
  • additional chambers (230, 250, 270) may be used to provide synthesis, purification, dialysis, compounding, and/or concentration of one or more therapeutic compositions; or to perform any other suitable function(s).
  • Fluid may be communicated from one chamber (230) to another chamber (230) via a fluidic connector (232).
  • fluidic connector (232) is operable like a valve between an open and closed state (e.g., similar to valve chamber (224)).
  • fluidic connector (232) remains open throughout the process of making the therapeutic composition.
  • chambers (230) are used to provide synthesis of polynucleotides, though chambers (230) may alternatively serve any other suitable purpose(s).
  • another valve chamber (234) is interposed between one of chambers (230) and one of chambers (250), such that fluid may be selectively communicated from chamber (230) to chamber (250).
  • Chambers (250) are provided in a pair and are coupled with each other such that process chip (200) may communicate the fluid back and forth between chambers (250). While a pair of chambers (250) are provided in the present example, any other suitable number of chambers (250) may be used, including just one chamber (250) or more than two chambers (250). Chambers (250) may be used to provide purification of the fluid and/or may serve any of the other various purposes described herein; and may have any suitable configuration.
  • chamber (250) may include a material that is configured to absorb selected moieties from a fluidic mixture in chamber (250).
  • the material may include a cellulose material, which may selectively absorb double-stranded mRNA from a mixture.
  • the cellulose material may be inserted in only one chamber (250) of a pair of chambers (250), such that upon mixing the fluid from the first chamber (250) of the pair to the second chamber (250), mRNA and/or some other component may be effectively removed from the fluidic mixture, which may then be transferred to another pair of chambers (270) further downstream for further processing or export.
  • chambers (250) may be used for any other suitable purpose.
  • valve chambers (252) are interposed between each chamber (250) and a corresponding chamber (270), such that fluid may be selectively communicated from chambers (250) to chambers (270) via valve chambers (252). Chambers (270) are also coupled with each other such that process chip (200) may communicate the fluid back and forth between chambers (270). Chambers (270) may be used to provide mixing of the fluid and/or may serve any of the other various purposes described herein; and may have any suitable configuration.
  • chambers (270) are also coupled with additional fluid ports (221) via corresponding fluid channels (223) and valve chambers (225).
  • Fluid ports (221), fluid channels (223), and valve chambers (225) may be configured and operable like fluid ports (220), fluid channels (222), and valve chambers (224) described above.
  • fluid ports (221) are used to communicate additional fluids to chambers (270).
  • fluid ports (221) may be used to communicate fluid from process chip (200) to another device. For instance, fluid from chambers (270) may be communicated via fluid ports (221) directly to another process chip (200), to one or more vials in reagent storage frame (107), or elsewhere.
  • Process chip (200) further includes several reservoir chambers (260).
  • each reservoir chamber (260) is configured to receive and store fluid that is being communicated to or from a corresponding chamber (250, 270).
  • Each reservoir chamber (260) has a corresponding inlet valve chamber (262) and outlet valve chamber (264).
  • Each inlet valve chamber (262) is interposed between reservoir chamber (260) and the corresponding chamber (250, 270) and is thereby operable to permit or prevent the flow of fluid between reservoir chamber (260) and the corresponding chamber (250, 270).
  • Each outlet valve chamber (264) is operable to meter the flow of fluid between reservoir chamber (260) and a corresponding fluid port (266).
  • each fluid port (266) is configured to communicate fluid from a corresponding vial in reagent storage frame (107) to a corresponding reservoir chamber (260).
  • each fluid port (266) may be configured to communicate fluid from a corresponding reservoir chamber (260) to a corresponding vial in reagent storage frame (107).
  • reservoir chambers (260) are used to provide metering of fluid communicated to and/or from process chip (200).
  • reservoir chambers (260) may be utilized for any other suitable purposes, including but not limited to pressurizing fluid that is communicated to and/or from process chip (200).
  • process chip (200) of this example includes a plurality of pressure ports (240).
  • Each pressure port (240) has an associated pressure channel (244) formed in process chip (200), such that pressurized gas communicated through pressure port (240) will be further communicated through the corresponding pressure channel (244).
  • each pressure port (240) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109).
  • each pressure channel (244) leads to a corresponding chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) to thereby provide valving or peristaltic pumping via such chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) as described in greater detail below.
  • Process chip (200) may also include electrical contacts, pins, pin sockets, capacitive coils, inductive coils, or other features that are configured to provide electrical communication with other components of system (100).
  • process chip (200) includes an electrically active region (212) that includes such electrical communication features. Electrically active region (212) may further include electrical circuits and other electrical components. In some versions, electrically active region (212) may provide communication of power, data, etc. While electrically active region (212) is shown in one particular location on process chip, electrically active region (212) may alternatively be positioned at any other suitable location or locations. In some versions, electrically active region (212) is omitted.
  • process chip (200) further includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306).
  • elastic layer (302) are in the form of a flexible membrane.
  • First plate (300) has an upper surface (210) and a lower surface (310), with lower surface (310) apposing elastic layer (302).
  • Second plate (304) has an upper surface (312) and a lower surface (314), with upper surface (312) apposing elastic layer (302); and with lower surface (314) apposing third plate (306).
  • Elastic layer (302) is thus interposed between first and second plates (300, 304).
  • another elastic layer (316) is also interposed between second and third plates (304, 306), though this elastic layer (316) is optional.
  • Plates (300, 304, 306) of the present example are substantially translucent to visible light and/or ultraviolet light.
  • substantially translucent is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a translucent material.
  • the one or more of plates (300, 304, 306) may comprise materials that are substantially transparent to visible light and/or ultraviolet light.
  • substantially transparent is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a completely transparent material.
  • one or more of plates may provide transmission of ultraviolet light at a wavelength of approximately 260 nm at a transmission rate ranging from approximately 0.2% to approximately 20%, including from approximately 0.4% to approximately 15%, or including from approximately 0.5% to approximately 10%.
  • Plates (300, 304, 306) of the present example are also rigid. In some other versions, one or more of plates (300, 304, 306) are semi-rigid. Plates (300, 304, 306) may comprise glass, plastic, silicone, and/or any other suitable material(s). In some versions, one or more of plates (300, 304, 306) is formed as a lamination of two or more layers of material, such that each plate (300, 304, 306) does not necessarily need to be formed as a single homogenous continuum of material. The material(s) comprising one of plates (300, 304, 306) may also differ from the material(s) comprising other plates (300, 304, 306).
  • Elastic layer (302) of the present example is formed as a liquid-impermeable flexible membrane.
  • elastic layer (302) is gas-permeable despite being liquid-impermeable.
  • certain regions of elastic layer (302) are treated to be gas-permeable while the non-treated regions of elastic layer (302) are gas- impermeable.
  • elastic layer (302) may be used to drive fluids across process chip (200) via peristaltic pumping action.
  • elastic layer (302) may be used to provide valves at various locations along process chip (200).
  • a single sheet of elastic material spans across the width of process chip (200) to form elastic layer (302).
  • elastic layer (302) may include a membrane comprising polydimethylsilicone (PDMS) elastomer film.
  • PDMS polydimethylsilicone
  • first and second plates (300, 304) cooperate to define a plurality of chambers (320, 322, 324, 326), with elastic layer (302) bisecting each chamber (320, 322, 324, 326) into a corresponding upper chamber region (330) and lower chamber region (332).
  • Chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) shown in FIG. 3 may be configured and operable just like chambers (320, 322, 324, 326) shown in FIGS. 4A-4F.
  • chamber (320) may be analogous to chamber (264)
  • chamber (322) may be analogous to chamber (260)
  • chamber (324) may be analogous to chamber (262)
  • chamber (326) may be analogous to chamber (250).
  • fluid port (220) is formed through first plate (300).
  • a corresponding opening (342) is formed through the region of elastic layer (302) underlying fluid port (220).
  • Fluid channel (222) extends from opening (342) to lower chamber region (332) of first chamber (320).
  • fluid port (220) is configured to receive a fluid line (206) from fluid interface assembly (109). The distal end of fluid line (206) is configured to seal against the region of elastic layer (302) that is exposed by fluid port (220) and communicate fluid (207) through opening (342).
  • a spring or other resilient member provides a resilient bias to fluid line (206), urging the distal end of fluid line (206) against the region of elastic layer (302) that is exposed by fluid port (220) to thereby maintain the seal.
  • Fluid (207) from fluid line (206) reaches lower chamber region (332) of first chamber (320) via fluid channel (222). As described in greater detail below, this fluid (207) may be further communicated from first chamber (320) to other chambers (322, 324, 326) through a peristaltic pumping action that is provided via elastic layer (302). After reaching fourth chamber (326), the fluid (207) may be further communicated to other chambers or other features in process chip (200), may be communicated to a storage vial in reagent storage frame (107), or may be otherwise processed. The path for fluid (207) thus does not necessarily terminate at fourth chamber (326). It should also be understood that any of the other fluid ports (221, 266) shown in FIG. 3 may be configured and operable like fluid port (220) shown in FIGS. 4A-4F.
  • Pressure port (240) is formed through first plate (300).
  • a corresponding opening (344) is formed through the region of elastic layer (302) underlying pressure port (240).
  • Pressure channel (244) extends from opening (344) to upper chamber region (330) of first chamber (320).
  • pressure port (240) is configured to receive a pressure line (208) from fluid interface assembly (109), to thereby receive pressurized gas from pressure source (117).
  • the distal end of pressure line (208) is configured to seal against the region of elastic layer (302) that is exposed by pressure port (240) and communicate either positively pressurized gas or negatively pressurized gas through opening (344).
  • a spring or other resilient member provides a resilient bias to pressure line (208), urging the distal end of pressure line (208) against the region of elastic layer (302) that is exposed by pressure port (240) to thereby maintain the seal.
  • Positively pressurized gas or negatively pressurized gas from pressure line (208) reaches upper chamber region (330) of fourth chamber (326) via pressure channel (244).
  • FIGS. 4A-4F depict just one pressure line (208) being coupled with process chip (200), process chip (200) may have several coupled pressure lines (208), with such pressure lines (208) independently applying positive or negative pressure to corresponding chambers (320, 322, 324, 326) of process chip (200).
  • one or more of chambers (320, 322, 324, 326) has its own dedicated pressure line (208) and corresponding pressure channel (244).
  • one or more of chambers (320, 322, 324, 326) may share a common pressure line (208), via the same pressure channel (244) or via separate pressure channels (244). While FIGS.
  • FIG. 4A-4F depict pressure channel (244) formed through second plate (304), some pressure channels (244) (or regions of pressure channels (244)) may be formed by first plate (300). For instance, some pressure channels (244) (or regions of pressure channels (244)) may be formed between a recess in the lower surface of first plate (300) and the top surface of elastic layer (302).
  • elastic layer (302) may be operated to drive fluid through process chip (200) through a peristaltic pumping action; and to arrest movement of fluid through process chip (200) by providing a valving action.
  • chambers (320, 324) serve as valve chambers, while chamber (322) serves as a metering chamber.
  • Chamber (326) serves as a working chamber, such that synthesis, purification, dialysis, compounding, concentration, or some other process is performed in chamber (326).
  • This configuration, arrangement, and usage of chambers (320, 322, 324, 326) is provided as an illustrative example. Chambers (320, 322, 324, 326) may alternatively be configured, arranged, and used in other ways.
  • FIG. 4A shows process chip (200) in a state where fluid is not yet being communicated to process chip (200); and pressurized gas is not yet being communicated to process chip (200).
  • FIG. 4B positively pressurized gas is communicated to upper chamber region (330) of chamber (324), negatively pressurized gas is communicated to upper regions (330) of chambers (320, 322), and fluid (207) is communicated to chambers (320, 322).
  • the positively pressurized gas deforms the portion of elastic layer (302) in chamber (324) such that elastic layer (302) seats against the surface of lower chamber region (332) of chamber (324).
  • the positively pressurized gas deforms the portion of elastic layer (302) in chamber (320) such that elastic layer (302) seats against the surface of lower chamber region (332) of chamber (320).
  • This seating of elastic layer (302) against the surface of lower chamber region (332) of chamber (320) drives the fluid (207) out from chamber (320) and results in chamber (320) operating like a closed valve in the state shown in FIG. 4C.
  • the volume of fluid (207) in chamber (322) is unaffected in the state shown in FIG. 4C.
  • Chamber (322) may thus be used to provide metering of fluid (207), such that only a precise, predetermined volume of fluid (207) is communicated further along process chip (200).
  • metered volumes may be on the order of approximately 10 nL, 20 nL, 25 nL, 50 nL, 75 nL, 100 nL, 1 microliter, 5 microliters, etc.
  • chamber (324) Since the deformed portion of elastic layer (302) in chamber (324) is effectively sealing off chamber (324) from chamber (322) (e.g., such that chamber (324) is operating like a valve in a closed state), fluid (207) travels from chamber (324) into chamber (326).
  • fluid (207) has been evacuated from chambers (320, 332, 324), and chamber (326) contains the volume of fluid (207) that was precisely metered in chamber (322).
  • Fluid (207) in chamber (326) may be further processed within chamber (326) in accordance with the teachings herein.
  • fluid (207) in chamber (326) may be communicated to one or more other chambers in process chip (200), may be communicated to a vial in reagent storage frame (107), or may be otherwise handled.
  • fluid (207) was communicated along chambers (320, 322, 324), in a sequence, to reach chamber (326) via a peristaltic action created through elastic layer (302) in response to positively pressurized gas or negatively pressurized gas being communicated to upper chamber regions (330) of chambers (320, 322, 324, 326) in a particular sequence.
  • peristaltic pumping may have particular advantage for moving fluid that may be viscous or contain suspended particles such as purification or capture beads.
  • Such peristaltic pumping through selective deformation of elastic layer (302) may also be referred to as pneumatic barrier deflection or “pneumodeflection.”
  • process chip (200) may include one or more chambers that are configured to provide ventilation of a fluid pathway or otherwise evacuate gas from the fluid pathway.
  • ventilation or evacuation may be performed as part of a priming process as fluid is initially introduced to process chip (200).
  • ventilation or evacuation may be performed to relieve gas that is generated in the fluid during the process of forming the therapeutic composition.
  • Ventilation or gas relief chambers may be referred to as “vacuum caps.”
  • at least the region of elastic layer (302) that is positioned in the vacuum cap is gas permeable (while still being liquid impermeable).
  • Negatively pressurized gas may be applied to the upper chamber region (330) of the chamber that is being used as a vacuum cap, and this negatively pressurized gas may draw the air or gas from the fluid pathway out through the corresponding region of elastic layer (302).
  • the upper chamber region (330) of the chamber that is being used as a vacuum cap includes one or more projections or stand-off features that prevent the corresponding region of elastic layer (302) from fully seating against the surface of the upper chamber region (330) of the chamber that is being used as a vacuum cap. This may further promote evacuation of air or other gas via the vacuum cap.
  • process chip (200) it may be desirable to provide a version of process chip (200) that provides substantially efficient heat conduction through at least a portion of process chip (200).
  • a version of process chip (200) that is configured to provide template synthesis with one or more PCR chambers, it may be desirable to heat those PCR chambers to facilitate the PCR process.
  • polymeric materials may provide suitable heat conduction to achieve acceptable PCR results
  • other materials such as metals, may provide improved PCR results (e.g., greater efficiency) by providing improved heat conduction.
  • metal may also facilitate the expansion of the volume defined by each PCR chamber, which may in turn further provide greater throughput in PCR processes.
  • elastic layer (302) when elastic layer (302) is pneumatically deformed against a surface of a chamber (320, 322, 324, 326) to drive fluid out of that chamber (320, 322, 324, 326), there may be a tendency for elastic layer (302) to trap pockets of fluid against the floor and/or sidewall of chamber (320, 322, 324, 326), such that not all of the fluid exits chamber (320, 322, 324, 326) when elastic layer (302) is pneumatically deformed to drive fluid out of that chamber (320, 322, 324, 326). It may therefore be desirable to provide a feature in a chamber that prevents an elastic layer from trapping pockets of fluid within a chamber when the elastic layer is pneumatically deformed to drive fluid out of that chamber.
  • beads may be present within one or more chambers (320, 322, 324, 326). For instance, in a process chip (200) that has one or more chambers where PCR is performed, beads may be present within the fluid in such chambers.
  • elastic layer (302) is pneumatically deformed against a bottom surface of the PCR chamber to drive fluid out of the PCR chamber, it may be desirable to provide a region in the PCR chamber where the beads may accumulate, such that elastic layer (302) will not press against those beads.
  • FIGS. 5-8 show an example of a process chip (400) that may provide at least some, if not all, of the features and functionalities described above.
  • Process chip (400) is similar to process chip (200) described above, except as otherwise described below.
  • process chip (400) may be utilized to provide in-vitro synthesis of templates, including but not limited to therapeutic polynucleotides templates.
  • process chip (400) may be utilized in other processes.
  • process chip (400) of this example includes a plurality of fluid ports (410) and a plurality of pressure ports (420). Each fluid port (410) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109). Each pressure port (420) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109). As shown in FIG. 7, process chip (400) of this example further includes a plurality of PCR chambers (450), in which PCR amplification may be performed as part of a process of forming a template for a therapeutic polynucleotide. While four PCR chambers (450) are shown, process chip (400) may have any other suitable number of PCR chambers (450).
  • process chip (400) of this example further includes a first layer (500), a second layer (900), a third layer (600), a fourth layer (700), a fifth layer (750), and a sixth layer (800).
  • layers (500, 600, 700, 800, 900), and their structural and functional relationships with each other, will be described in greater detail below.
  • fluid ports (410) and pressure ports (420) are all positioned on a top region of process chip (400), at least within the frame of reference of FIG. 5.
  • fluid ports (410) are all positioned on a top region of process chip (400) while pressure ports (420) are all positioned on a bottom region of process chip (400).
  • fluid ports (410) are all positioned on a bottom region of process chip (400) while pressure ports (420) are all positioned on a top region of process chip (400).
  • some fluid ports (410) may be positioned on a top region of process chip (400) while other fluid ports (410) are positioned on a bottom region of process chip (400).
  • some pressure ports (420) may be positioned on a top region of process chip (400) while other pressure ports (420) are positioned on a bottom region of process chip (400).
  • FIGS. 9-11 show first layer (500) in greater detail.
  • First layer (500) is in the form of a rigid plate in this example.
  • first layer (500) may be substantially translucent to visible light and/or ultraviolet light.
  • substantially translucent is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a translucent material.
  • First layer (500) may comprise glass, plastic, silicone, and/or any other suitable material(s).
  • first layer (500) is formed as a lamination of two or more layers of material, such that first layer (500) does not necessarily need to be formed as a single homogenous continuum of material.
  • first layer (500) includes an upper surface (502), a lower surface (504), first plurality of openings (510) formed through both surfaces (502, 504), and a second plurality of openings (520) formed through both surfaces (502, 504).
  • Openings (510) form part of fluid ports (410), such that each fluid port (410) has a respective opening (510), and such that openings (510) are configured to accommodate fluid lines (206).
  • Openings (520) form part of pressure ports (420), such that each pressure port (420) has a respective opening (520), and such that openings (520) are configured to accommodate pressure lines (208).
  • a plurality of PCR chamber portions (552) are formed as recesses in lower surface (504), providing protruding regions (550) in upper surface (502). Each PCR chamber portion (552) forms a “dry” region of a respective PCR chamber (450) as described herein.
  • lower surface (504) of first layer (500) further defines a plurality of pneumatic channels (522), which are formed as recesses in lower surface (504).
  • Each pneumatic channel (522) is in pneumatic communication with a corresponding opening (520).
  • Each pneumatic channel (522) is further configured to define a space between first layer (500) and second layer (900), such that pressurized gas may be communicated along pneumatic channels (522).
  • Lower surface (504) of first layer (500) further defines a plurality of valve chamber portions (530) and a plurality of pump chamber portions (540).
  • Valve chamber portions (530) and pump chamber portions (540) are formed as recesses in lower surface (504); and are in pneumatic communication with corresponding pneumatic channels (522). Pressurized gas may thus be communicated from pressure ports (420) to valve chamber portions (530) and pump chamber portions (540) via respective pneumatic channels (522).
  • Valve chamber portions (530) form “dry” regions of valve chambers as described herein; while pump chamber portions (540) form “dry” regions of pump chambers as described herein.
  • valve chamber portions (530) have a circular shape while pump chamber portions (540) have an oblong or stadium shape, though other shapes may be used.
  • valve chambers associated with valve chamber portions (530) may be operated like chambers (320, 322) described above; while pump chambers associated with pump chamber portions (540) may be operated like chambers (322, 326) described above.
  • each chamber portion (530, 540) has its own associated pneumatic channel (522) and pressure port (520) in this example
  • each PCR chamber portion (552) also has its own associated pneumatic channel (522) and pressure port (520) in this example.
  • each chamber portion (530, 540, 552) may be pneumatically pressurized independently of the other chamber portions (530, 540, 552).
  • Second layer (900) is interposed between first layer (500) and third layer (600).
  • Second layer (900) of this example is in the form of a flexible membrane.
  • second layer (900) comprises polydimethylsilicone (PDMS) elastomer film.
  • PDMS polydimethylsilicone
  • any other suitable material(s) may be used to form second layer (900).
  • Second layer (900) may be configured and operable like elastic layer (302) described above.
  • Second layer (900) may thus pass through chambers collectively defined by layers (500, 800) to separate such chambers into an upper chamber region (which receives pneumatic pressure) and a lower chamber region (which receives fluid).
  • Such chambers include PCR chambers (450), valve chambers as described herein, and pump chambers as described herein.
  • Second layer (900) may further thus be pneumatically deformed to provide valving, peristaltic pumping, etc. within process chip (400).
  • second layer (900) may include a plurality of openings formed therethrough. For instance, such openings may be positioned at fluid ports (410), under openings (510), to allow fluid to be communicated through second layer (900) at fluid ports (410).
  • each fluid port (410) is configured to receive a fluid line (206) from fluid interface assembly (109). The distal end of fluid line (206) may be configured to seal against the region of second layer (900) that is exposed by fluid port (410) and communicate fluid through the respective openings in second layer (900).
  • a spring or other resilient member provides a resilient bias to fluid line (206), urging the distal end of fluid line (206) against the region of second layer (900) that is exposed by fluid port (410) to thereby maintain the seal. Fluid from fluid line (206) may be further communicated through process chip (400) as described in greater detail below.
  • each chamber in process chip (400) that is above second layer (900) constitutes a “dry” chamber region since this region receives pressurized gas to pneumatically deflect second layer (900).
  • the region of each chamber in process chip (400) that is below second layer (900) constitutes a “wet” chamber region since this region receives fluid.
  • process chip (400) may provide pressurized gas to lower region of each chamber, such that the lower chamber region constitutes a “dry” chamber region; while the upper chamber region receives fluid, such that upper chamber constitutes a “wet” chamber region.
  • FIGS. 12-14 show third layer (600) in greater detail.
  • Third layer (600) is in the form of a rigid plate in this example.
  • third layer (600) is substantially translucent to visible light and/or ultraviolet light.
  • Third layer (600) may comprise glass, plastic, silicone, and/or any other suitable material(s).
  • third layer (600) is formed as a lamination of two or more layers of material, such that third layer (600) does not necessarily need to be formed as a single homogenous continuum of material.
  • third layer (600) includes an upper surface (602), a lower surface (604), a first plurality of openings (610) formed through both surfaces (602, 604), and a second plurality of openings (650) formed through both surfaces (602, 604).
  • Some of openings (610) form part of pressure ports (420), such that each pressure port (420) has a respective opening (610), and such that openings (610) are configured to allow communication of pressurized gas through third layer (600).
  • corresponding openings may be formed through second layer (900), such that these openings in second layer (900) are between openings (520) of first layer (500) and at least some of openings (610) of third layer (600).
  • Openings (650) are positioned in part of the “wet” region of PCR chambers (450) in this example.
  • Each opening (650) has a sidewall (652) that extends from upper surface (602) to lower surface (604).
  • upper surface (602) of third layer (600) further defines a plurality of fluid channels (612), which are formed as recesses in upper surface (602).
  • Each fluid channel (612) is in fluid communication with a corresponding opening (610).
  • Each fluid channel (612) is further configured to define a space between third layer (600) and second layer (900), such that fluid may be communicated along fluid channels (612).
  • Upper surface (602) of third layer (600) further defines a plurality of valve chamber portions (630) and a plurality of pump chamber portions (640).
  • Valve chamber portions (630) and pump chamber portions (640) are formed as recesses in upper surface (602); and are in fluid communication with corresponding fluid channels (612). Fluid may thus be communicated from fluid ports (410) to valve chamber portions (630) and pump chamber portions (640) via respective openings (610) and fluid channels (612).
  • Valve chamber portions (630) form “wet” regions of valve chambers as described herein (opposite to a corresponding valve chamber portion (530)); while pump chamber portions (640) form “wet” regions of pump chambers (opposite to a corresponding pump chamber portion (540)) as described herein.
  • valve chamber portions (630) have a circular shape while pump chamber portions (640) have an oblong or stadium shape, though other shapes may be used.
  • valve chambers associated with valve chamber portions (630) may be operated like chambers (320, 322) described above; while pump chambers associated with pump chamber portions (640) may be operated like chambers (322, 326) described above.
  • Each chamber portion (630, 640) has its own associated fluid channel (612) and opening (610) in this example. Thus, each chamber portion (630, 640) may receive fluid independently of the other chamber portions (630, 640).
  • Each opening (650) also has a respective fluid channel (612) leading thereto, such that fluid may be communicated to each PCR chamber (450) via the fluid channel (612) leading to the opening (650) associated with the PCR chamber (450).
  • a pump chamber portion (640) and set of valve chamber portions (630) is interposed between an opening (610) and each fluid channel (612) leading to each respective opening (650). The pumps and valves defined by these portions (640, 630) may thus drive and regulate the communication of fluid to each PCR chamber (450).
  • third layer (600) includes a plurality of bridging channels (660) and sidewall channels (654).
  • Each bridging channel (660) is formed as a recess in upper surface (602).
  • Each sidewall channel (654) is formed as a recess in a corresponding sidewall (652).
  • each bridging channel (660) and the pair of adjacent sidewall channels (654) are configured to provide a path for communication of fluid from one PCR chamber (450) to an adjacent PCR chamber (450).
  • third layer (600) further includes a plurality of fluid channels (614) on lower surface (604). Fluid channels (614) are formed as recesses on lower surface (604). Each fluid channel (614) is further configured to define a space between third layer (600) and fourth layer (700), such that fluid may be communicated along fluid channels (614). An opening (610) is positioned at each end of each fluid channel (614).
  • Openings (610) and fluid channels (614) thus cooperate to allow fluid to be communicated from one region at upper surface (602) of third layer (600) to another region at upper surface (602) of third layer (600) via lower surface (604) of third layer (600), which may facilitate routing of fluid more easily than could otherwise be achieved if the fluid were only routed via fluid channels (612) in upper surface (602).
  • the inclusion of fluid channels (614) on lower surface (604) may allow fluid to be routed along a path that would otherwise intersect the fluid path provided via a fluid channel (612) if such path were moved from lower surface (604) to upper surface (602).
  • FIG. 15 shows fourth layer (700) in greater detail.
  • Fourth layer (700) may include an elastomeric membrane, such as silicone and/or any other suitable material(s).
  • fourth layer (700) of this example includes a plurality of openings (702). Openings (702) are positioned in part of the “wet” region of PCR chambers (450) in this example. Openings (702) of fourth layer (700) are thus sized and positioned to correspond with the size and position of openings (650) of third layer (600).
  • Fourth layer (700) further includes a set of notches (704) at each opening (650). Notches (704) are sized and positioned to correspond with the size and position of sidewall channels (654), such that each sidewall channel (654) has an adjacent notch (704).
  • fifth layer (750) is not shown separately, it should be understood that fifth layer (750) may have a structural configuration that is identical to the structural configuration of fourth layer (700) as shown in FIG. 15 and described above.
  • layers (700, 750) together provide a gasket forming a seal between adjacent regions of third layer (600) and eighth layer (800).
  • one of layers (700, 750) is omitted.
  • FIGS. 16-18 show sixth layer (800) in greater detail.
  • Sixth layer (800) of this example comprises a metal material.
  • sixth layer (800) may comprise aluminum or titanium.
  • any other suitable material(s) may be used to form sixth layer (800), including non-metallic materials (e.g., ceramic materials, thermally conductive polymers, etc.).
  • sixth layer (800) may include one or more coatings (e.g., hexamethyldisiloxane (HMDSO), polytetrafluoroethylene (PTFE), etc.).
  • one or more heating elements is/are integrated within sixth layer (800) and/or on sixth layer (800).
  • Sixth layer (800) includes an upper surface (802) and a lower surface (804).
  • a plurality of PCR chamber portions (850) are formed as recesses in upper surface (802), providing protruding regions (852) in lower surface (804).
  • Each PCR chamber portion (850) forms a “wet” region of a respective PCR chamber (450) as described herein.
  • a lip (854) protrudes upwardly from upper surface (802) around each PCR chamber portion (850). Lips (854) may provide concentrated compression against layers (700, 750) when process chip (400) is assembled.
  • each lip (854) surrounds an outer perimeter around each PCR chamber portion (850), the concentrated compression provided by lips (854) against layers (700, 750) may minimize the risk of fluid leaking from PCR chambers (450) during use of process chip (400). Additional lips (856) are provided on other regions of upper surface (802); and also provide concentrated compression against layers (700, 750) when process chip (400) is assembled.
  • a screw is used to secure eighth layer (800) to first layer (500) and compress other layers (600, 700, 750, 900) therebetween.
  • any other suitable structures or techniques may be used to secure layers (500, 600, 700, 750, 800, 900) together, including but not limited to clips, clamps, adhesives, epoxies, etc.
  • each PCR chamber portion (850) includes a floor (860), a sidewall (870), and a shelf (880) that is elevated relative to floor (860).
  • a pair of sidewall channels (872) are defined in each sidewall (870).
  • Each pair of sidewall channels (872) is positioned to correspond with the position of notches (704) and sidewall channels (654), such that each sidewall channel (872) has an adjacent notch (704); and such that notches (704) are positioned between corresponding adjacent sidewall channels (654, 872).
  • each sidewall channel (872) extends from floor (860) to notch (704), with notch (704) providing a path for fluid communication from sidewall channel (872) to sidewall channel (654), and with bridging channel (660) further extending the path for fluid communication from sidewall channel (654).
  • Another sidewall channel (654) is positioned at the other end of bridging channel (660), leading to another notch (704) and sidewall channel (872) in another PCR chamber portion (850).
  • channels (654, 660, 872) and notches (704) cooperate to provide a path for fluid communication from one PCR chamber (450) to an adjacent PCR chamber (450).
  • FIGS. 22A-22B show a series of operational states where fluid may be communicated from one PCR chamber (450b) to an adjacent PCR chamber (450a) via channels (654, 660, 872) and notches (704).
  • FIG. 22A shows process chip (400) in an operational state where the “dry” regions of the depicted PCR chambers (450a, 450b) (i.e., above second layer (900)) are not receiving pressurized gas, such that second layer (900) is substantially flat; and such that fluid may be held in one or more of the “wet” regions of the depicted PCR chambers (450a, 450b) (i.e., below second layer (900)).
  • FIG. 22B shows process chip (400) in an operational state where the “dry” region of PCR chamber (450b) receives pressurized gas, which causes the region of second layer (900) to deform downwardly into the “wet” region of PCR chamber (450b).
  • This deformation of second layer (900) reduces the volume of the “wet” region of PCR chamber (450b), which drives the fluid from the “wet” region of the associated PCR chamber (450b) to adjacent PCR chamber (450a) via channels (654, 660, 872) and notches (704).
  • the other PCR chamber (450) (not shown in FIGS. 22A-22B) that is adjacent to PCR chamber (450b) also receives fluid from PCR chamber (450b) via corresponding channels (654, 660, 872) and notches (704) in the operational state shown in FIG. 22B.
  • Each PCR chamber (450) may define a capacity between floor (860), sidewall (870), and the underside of second layer (900); with a depth of each PCR chamber portion (850) being defined between upper surface (802) of sixth layer (800) and floor (860).
  • each PCR chamber (450) may have a capacity of at least approximately 0.3 mL; of up to approximately 33.2 mL; ranging from approximately 0.3 mL to approximately 33.2 mL; or ranging from approximately 1 mL to approximately 25 mL; or ranging from approximately 5 mL to approximately 20 mL.
  • each PCR chamber (450) may have a capacity of approximately 13.1 mL.
  • each PCR chamber (450) may have any other suitable capacity.
  • Each PCR chamber portion (850) may have a depth of at least approximately 0 mm; of up to approximately 27.1 mm; ranging from approximately 0 mm to approximately 27.1 mm; or ranging from approximately 1 mm to approximately 20 mm; or ranging from approximately 3 mm to approximately 12 mm.
  • each PCR chamber portion (850) may have a depth of approximately 7.4 mm.
  • each PCR chamber portion (850) may have any other suitable depth.
  • second layer (900) when second layer (900) is pneumatically deformed downwardly into the “wet” region of a PCR chamber (450), such as in the configuration shown in FIG. 22B, second layer (900) may contact floor (860) and/or sidewall (870) of PCR chamber portion (850) of that PCR chamber (450). In some such cases, if this contact occurred in the absence of channels (872), one or more localized regions of deformed second layer (900) may tend to trap pockets of fluid against floor (860) and/or sidewall (870) of PCR chamber portion (850), such that the trapped fluid does not exit PCR chamber (450).
  • channels (872) may provide a pathway for the fluid to escape PCR chamber (450) when second layer (900) is deformed downwardly toward floor (860), such that channels (872) prevent pockets of fluid from being trapped between localized regions of deformed second layer (900) and floor (860) and/or sidewall (870) (or otherwise reduce the risk of such pockets being formed).
  • FIGS. 23A-23B show process chip (400) in an operational state where the “dry” region of the PCR chamber (450b) (i.e., above second layer (900)) is not receiving pressurized gas, such that second layer (900) is substantially flat; and such that a fluid containing beads (1000) is held in the “wet” region of PCR chamber (450b) (i.e., below second layer (900)).
  • FIG. 23B shows process chip (400) in an operational state where the “dry” region of PCR chamber (450b) receives pressurized gas, which causes the region of second layer (900) to deform downwardly into the “wet” region of PCR chamber (450b).
  • This deformation of second layer (900) reduces the volume of the “wet” region of PCR chamber (450b), which drives the fluid from the “wet” region of the associated PCR chamber (450b) to one or more adjacent PCR chambers (450) via channels (654, 660, 872) and notches (704) as described above.
  • channels (654, 660, 872) and notches (704) as described above.
  • beads (1000) accumulate within a space (882) defined between shelf (880) of eighth layer (800) and an overhanging portion (606) of third layer (600).
  • process chip (400) prevents beads (1000) from being driven from PCR chamber (450b) to an adjacent PCR chamber (450a, 450c).
  • channels (872) have been described above as being formed in sidewall (870) of PCR chamber portion (850), other portions of process chip (400) may also have channels (872) formed in sidewalls. Such other portions of process chip (400) may provide various other functions, such that the utility of channels in sidewalls of recesses is not necessarily limited to PCR processes.
  • FIGS. 24-26 illustrate how channel portions (914, 964) in sidewalls (912, 962) may be integrated into a pump chamber portion (910) of a fluid inlet region (901) leading into a PCR chamber (450); and into a pump chamber portion (960) of a fluid outlet region (950) leading out of a PCR chamber (450).
  • fluids may also be communicated to PCR chamber (450) via one or more other routes; and that fluids may also be communicated from PCR chamber (450) via one or more other routes.
  • different PCR chambers (450) within the same process chip (400) may receive fluids via differently configured and/or positioned fluid inlet features; and fluids may be communicated from different PCR chambers (450) within the same process chip (400) via differently configured and/or positioned fluid outlet features.
  • a given PCR chamber (450) may receive fluids via additional fluid inlet features while also receiving fluids via fluid inlet region (901); and/or fluid may be communicated from a given PCR chamber (450) via additional fluid outlet features while also being communicated via fluid outlet region (950).
  • regions (901, 950) described below are just examples and are not intended to be limiting.
  • fluid inlet region (901) of this example includes a fluid input opening (902), a pair of valve chamber portions (906, 920), pump chamber portion (910), and a vacuum cap chamber (924).
  • a channel (904) extends from fluid input opening (902) to valve chamber portion (906).
  • a channel (908) extends from valve chamber portion (906) to pump chamber portion (910).
  • a channel (916) extends from pump chamber portion (910) to valve chamber portion (920).
  • a channel (922) extends from valve chamber portion (920) to vacuum cap chamber (924).
  • a channel (926) extends from vacuum cap chamber (924) to opening (650) of third layer (600).
  • Fluid input opening (902) represents an example of one of openings (610) described above, such that fluid inlet region (901) receives fluid via fluid input opening (902).
  • Each valve chamber portion (906, 920) represents a respective example of one of valve chamber portions (630) described above, such that second layer (900) may be deformed into each valve chamber portion (906, 920) to prevent flow of fluid.
  • Pump chamber portion (910) represents an example of one of pump chamber portions (640), such that second layer (900) may be deformed into pump chamber portion (910) to drive fluid out of pump chamber portion (910) and thereby provide metered flow of fluid.
  • pump chamber portion (910) includes a sidewall (912) providing a concave recess in upper surface (602) having a stadium shape.
  • a portion (914) of channel (916) is formed as a recess along sidewall (912).
  • Channel portion (914) extends to the lower region of pump chamber portion (910).
  • channel portion (914) may provide functionality similar to that described above with respect to sidewall channels (872). In other words, channel portion (914) may provide a path for fluid communication from the lower region of pump chamber portion (910) to the remainder of channel (916).
  • second layer (900) when second layer (900) is pneumatically deformed downwardly into pump chamber portion (910), second layer (900) may contact sidewall (912) of pump chamber portion (910). In some such cases, if this contact occurred in the absence of channel portion (914), one or more localized regions of deformed second layer (900) may tend to trap pockets of fluid against sidewall (912), such that the trapped fluid does not exit pump chamber portion (910).
  • the presence of channel portion (914) may provide a pathway for the fluid to escape pump chamber portion (910) when second layer (900) is deformed downwardly toward sidewall (912), such that channel portion (914) prevents pockets of fluid from being trapped between localized regions of deformed second layer (900) and sidewall (912) or otherwise reduces the risk of such pockets being formed.
  • channel portion (914) may also promote unidirectional flow of fluid through pump chamber portion (910) when second layer (900) is deformed downwardly toward sidewall (912). For instance, during some versions of operation, it may be desirable for fluid to enter pump chamber portion (910) only via channel (908); and for fluid to exit pump chamber portion (910) only via channel (916). In the absence of channel portion (914) along sidewall (912), there may be scenarios where some fluid may tend to exit pump chamber portion (910) via channel (908) when second layer (900) is deformed downwardly toward sidewall (912) (e.g., even if the valve provided by valve chamber portion (906) is in a closed state). However, in the present example, with channel portion (914) present along sidewall (912), the fluid may more reliably exit pump chamber portion (910) only via channel (916) when second layer (900) is deformed downwardly toward sidewall (912).
  • fluid outlet region (950) of this example includes a fluid output opening (952), a fluid input opening (972), a pair of valve chamber portions (976, 982), pump chamber portion (960), and a vacuum cap chamber (988).
  • a channel (954) extends from fluid output opening (952) to pump chamber portion (960).
  • a channel (974) extends from fluid input opening (972) to valve chamber portion (976).
  • a channel (978) extends from valve chamber portion (976) to pump chamber portion (960).
  • a channel (980) extends from pump chamber portion (960) to valve chamber portion (982).
  • a channel (986) extends from valve chamber portion (982) to vacuum cap chamber (988).
  • a channel (990) extends from vacuum cap chamber (988) to opening (650) of third layer (600).
  • a portion of (992) of channel (990) extends along the full height of sidewall (652), terminating at a lower surface (604) of third layer (600).
  • Each valve chamber portion (976, 982), pump chamber portion (960), and channel (954, 974, 978, 980, 986, 990) is formed as a recess in upper surface (602) of third layer (600).
  • Each opening (952, 972) represents a respective example of one of openings (610) described above, such that fluid exits fluid outlet region (950) via fluid output opening (952); and such that fluid enters fluid outlet region (950) via fluid input opening (972) (in addition to entering fluid outlet region (950) via channel (990) as described below).
  • Each valve chamber portion (976, 982) represents a respective example of one of valve chamber portions (630) described above, such that second layer (900) may be deformed into each valve chamber portion (976, 982) to prevent flow of fluid.
  • Pump chamber portion (960) represents an example of one of pump chamber portions (640), such that second layer (900) may be deformed into pump chamber portion (960) to drive fluid out of pump chamber portion (960) and thereby provide metered flow of fluid.
  • pump chamber portion (960) includes a sidewall (962) providing a concave recess in upper surface (602) having a stadium shape.
  • a portion (964) of channel (954) is formed as a recess along sidewall (962).
  • Channel portion (964) extends to the lower region of pump chamber portion (960).
  • channel portion (964) may provide functionality similar to that described above with respect to sidewall channels (872). In other words, channel portion (964) may provide a path for fluid communication from the lower region of pump chamber portion (960) to the remainder of channel (954).
  • second layer (900) when second layer (900) is pneumatically deformed downwardly into pump chamber portion (960), second layer (900) may contact sidewall (962) of pump chamber portion (960). In some such cases, if this contact occurred in the absence of channel portion (964), one or more localized regions of deformed second layer (900) may tend to trap pockets of fluid against sidewall (962), such that the trapped fluid does not exit pump chamber portion (960). However, the presence of channel portion (964) may provide a pathway for the fluid to escape pump chamber portion (960) when second layer (900) is deformed downwardly toward sidewall (962), such that channel portion (964) prevents pockets of fluid from being trapped between localized regions of deformed second layer (900) and sidewall (962) or otherwise reduces the risk of such pockets being formed.
  • channel portion (964) may also promote unidirectional flow of fluid through pump chamber portion (960) when second layer (900) is deformed downwardly toward sidewall (962). For instance, during some versions of operation, it may be desirable for fluid to enter pump chamber portion (960) only via channels (978); and for fluid to exit pump chamber portion (960) only via channel (954). In the absence of channel portion (964) along sidewall (962), there may be scenarios where some fluid may tend to exit pump chamber portion (960) via one or both of channels (978, 980) when second layer (900) is deformed downwardly toward sidewall (962) (e.g., even if the valves provided by valve chamber portions (976, 982) are in a closed state). However, in the present example, with channel portion (964) present along sidewall (962), the fluid may more reliably exit pump chamber portion (960) only via channel (954) when second layer (900) is deformed downwardly toward sidewall (962).
  • FIG. 26 shows additional features that may facilitate the communication of fluid from PCR chamber (450) to fluid outlet region (950) via portion of (992) of channel (990).
  • fourth layer (700) includes a notch (706) below portion of (992) of channel (990); and fifth layer (750) includes a notch (756) below notch (756).
  • a horizontal gap (871) is defined along the perimeter of the interface between the upper end of sidewall (870) and the lower surface of fifth layer (750). This gap (871) is in fluid communication with sidewall channels (872). Gap (871) is also in fluid communication with portion of (992) of channel (990) via notches (706, 756).
  • gap (871) and notches (706, 756) may cooperate with sidewall channels (872) to further promote communication of fluid from PCR chamber (450) to a fluid outlet region (950) via portion of (992) of channel (990).
  • channels may be provided in sidewalls of recesses anywhere within process chip (400) — to prevent (or reduce the risk of) formation of localized fluid pockets under deformed second layer (900), to promote unidirectional flow of fluid through a chamber defined by the recess, and/or for any other purpose.
  • sidewall channels (872) and channel portions (914, 964) are just examples.
  • Such sidewall channels and channel portions may be integrated into sidewalls of recesses that serve any other purposes, such that such sidewall channels and channel portions need not necessarily be limited to recesses forming chambers that are used to perform PCR processes; or chambers that are used to provide pumping, etc.
  • sidewall channels and channel portions may be provided in variations of process chip (400) that are used in non-PCR processes; and in variations of process chip (400) that lack any components formed of metal.
  • PCR chambers (450) are each substantially asymmetrical (e.g., vertically asymmetrical) relative to second layer (900), with upper PCR chamber portions (552) that are formed as recesses in lower surface (504) of first layer (500) being sized and shaped substantially differently from lower PCR chamber portions (850) that are formed as recesses in upper surface (802) of sixth layer (800), such that each upper process chamber portion (552) may be configured to hold a first volume of fluid and the corresponding lower process chamber portion (850) may be configured to hold a second volume of fluid substantially greater than the first volume of fluid.
  • process chamber portions (552, 850) may be configured and operable in accordance with at least some of the teachings of U.S. Pat. App. No. 63/681,918, entitled “Microfluidic Apparatus with Asymmetrical Chambers,” filed on August 12, 2024, the disclosure of which is incorporated by reference herein in its entirety.
  • second layer (900) may be in the form of first and second liquid-impermeable flexible membranes laminated to each other in a vertically stacked arrangement; and/or may be configured and operable in accordance with at least some of the teachings of International Pub. No. WO 2024/196908, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,” published on September 26, 2024, the disclosure of which is incorporated by reference herein in its entirety. It may therefore be desirable to provide a variation of process chip (400) that relocates bridging channels (660) such that fluid flowing along bridging channels (660) is isolated from (e.g., does not contact) second layer (900).
  • FIGS. 27-39 show an example of a process chip (1400) that may provide at least some, if not all, of the features and functionalities described above.
  • Process chip (1400) is similar to process chip (400) described above, except as otherwise described below.
  • process chip (1400) may be utilized to provide in-vitro synthesis of templates, including but not limited to therapeutic polynucleotides templates.
  • process chip (1400) may be utilized in other processes.
  • process chip (1400) of this example includes a plurality of fluid ports (1410) and a plurality of pressure ports (1420). Each fluid port (1410) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109). Each pressure port (1420) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109). As shown in FIG. 29, process chip (1400) of this example further includes a plurality of PCR chambers (1450), in which PCR amplification may be performed as part of a process of forming a template for a therapeutic polynucleotide. While four PCR chambers (1450) are shown, process chip (1400) may have any other suitable number of PCR chambers (1450).
  • process chip (1400) of this example further includes a first layer (1500), a second layer (1900), athird layer (1600), a fourth layer (1700), a fifth layer (1750), and a sixth layer (1800).
  • First, second, third, fourth, fifth, and sixth layers (1500, 1900, 1600, 1700, 1750, 1800) are similar to first, second, third, fourth, fifth, and sixth layers (500, 900, 600, 700, 750, 800) described above, respectively, except as otherwise described below.
  • Second layer (1900) is interposed between first layer (1500) and third layer (1600).
  • Second layer (1900) may be in the form of first and second liquid-impermeable flexible membranes laminated to each other in a vertically stacked arrangement.
  • second layer (1900) may be used to drive fluids across process chip (1400) via peristaltic pumping action.
  • second layer (1900) may be used to provide valves at various locations along process chip (1400).
  • one or both flexible membranes of second layer (1900) may comprise polydimethylsilicone (PDMS) elastomer film.
  • PDMS polydimethylsilicone
  • any other suitable material(s) may be used to form second layer (1900).
  • second layer (1900) may be configured and operable in accordance with at least some of the teachings of International Pub. No. WO 2024/196908, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,” published on September 26, 2024, the disclosure of which is incorporated by reference herein in its entirety; and/or U.S. Pat. No. 11,926,817, entitled “Microfluidic Apparatus and Methods of Use Thereof,” issued on March 12, 2024, the disclosure of which is incorporated by reference herein in its entirety.
  • FIGS. 31-33 show third layer (1600) in greater detail.
  • Third layer (1600) is in the form of a rigid plate in this example.
  • third layer (1600) is substantially translucent to visible light and/or ultraviolet light.
  • Third layer (1600) may comprise glass, plastic, silicone, and/or any other suitable material(s).
  • third layer (1600) is formed as a lamination of two or more layers of material, such that third layer (1600) does not necessarily need to be formed as a single homogenous continuum of material.
  • third layer (1600) includes an upper surface (1602), a lower surface (1604), a first plurality of openings (1610) formed through both surfaces (1602, 1604), and a second plurality of openings (1650) formed through both surfaces (1602, 1604).
  • Some of openings (1610) form part of pressure ports (1420), such that each pressure port (1420) has a respective opening (1610), and such that openings (1610) are configured to allow communication of pressurized gas through third layer (1600).
  • corresponding openings may be formed through second layer (1900), such that these openings in second layer (1900) are between openings (1520) of first layer (1500) and at least some of openings (1610) of third layer (1600).
  • Openings (1650) are positioned in part of the “wet” region of PCR chambers (1450) in this example.
  • Each opening (1650) has a sidewall (1652) that extends from upper surface (1602) to lower surface (1604).
  • upper surface (1602) of third layer (1600) further defines a plurality of fluid channels (1612), which are formed as recesses in upper surface (1602).
  • Each fluid channel (1612) is in fluid communication with a corresponding opening (1610).
  • Each fluid channel (1612) is further configured to define a space between third layer (1600) and second layer (1900), such that fluid may be communicated along fluid channels (1612).
  • Upper surface (1602) of third layer (1600) further defines a plurality of valve chamber portions (1630) and a plurality of pump chamber portions (1640).
  • Valve chamber portions (1630) and pump chamber portions (1640) are formed as recesses in upper surface (1602); and are in fluid communication with corresponding fluid channels (1612). Fluid may thus be communicated from fluid ports (1410) to valve chamber portions (1630) and pump chamber portions (1640) via respective openings (1610) and fluid channels (1612).
  • third layer (1600) includes a plurality of bridging channels (1660).
  • each bridging channel (1660) is formed as a recess in lower surface (1604).
  • each bridging channel (1660) is configured to provide a path for communication of fluid from one PCR chamber (1450) to an adjacent PCR chamber (1450).
  • third layer (1600) further includes a plurality of fluid channels (1614) on lower surface (1604). Fluid channels (1614) are formed as recesses on lower surface (1604).
  • Each fluid channel (1614) is further configured to define a space between third layer (1600) and fourth layer (1700), such that fluid may be communicated along fluid channels (1614).
  • An opening (1610) is positioned at each end of each fluid channel (1614). Openings (1610) and fluid channels (1614) thus cooperate to allow fluid to be communicated from one region at upper surface (1602) of third layer (1600) to another region at upper surface (1602) of third layer (1600) via lower surface (1604) of third layer (1600).
  • FIG. 34 shows fourth layer (1700) in greater detail.
  • Fourth layer (1700) may include an elastomeric membrane, such as silicone and/or any other suitable material(s).
  • fourth layer (1700) of this example includes a plurality of openings (1702). Openings (1702) are positioned in part of the “wet” region of PCR chambers (1450) in this example. Openings (1702) of fourth layer (1700) are thus sized and positioned to correspond with the size and position of openings (1650) of third layer (1600).
  • Fourth layer (1700) further includes a set of notches (1704) at each opening (1650). Notches (1704) are sized and positioned to correspond with the size and position of bridging channels (1660), such that each bridging channel (1660) has a pair of adjacent notches (1704).
  • FIG. 35 shows fifth layer (1750) in greater detail.
  • Fifth layer (1750) may include an elastomeric membrane, such as silicone and/or any other suitable material(s).
  • Fifth layer (1750) of this example includes a single opening (1752). Opening (1752) is positioned in part of the “wet” region of PCR chambers (1450) in this example. Opening (1752) of fifth layer (1750) is thus sized and positioned to correspond with the collective size and position of openings (1702) of fourth layer (1700).
  • Fifth layer (1750) further includes a set of notches (1754) at corresponding sides of opening (1752).
  • layers (1700, 1750) together provide a gasket forming a seal between adjacent regions of third layer (1600) and eighth layer (1800).
  • one of layers (1700, 1750) is omitted.
  • FIGS. 36-37 show sixth layer (1800) in greater detail.
  • Sixth layer (1800) includes an upper surface (1802) and a lower surface (1804).
  • a plurality of PCR chamber portions (1850) are formed as recesses in upper surface (1802), providing protruding regions (1852) in lower surface (1804).
  • Each PCR chamber portion (1850) forms a “wet” region of a respective PCR chamber (1450) as described herein.
  • inner and outer lips (1853, 1854) protrude upwardly from upper surface (1802) and cooperate with each other to substantially surround each PCR chamber portion (1850) while defining gaps (1855) that are positioned to correspond with the position of notches (1704) and bridging channels (1660).
  • Lips (1853, 1854) may provide concentrated compression against layers (1700, 1750) when process chip (1400) is assembled. Since lips (1853, 1854) substantially surround an outer perimeter around each PCR chamber portion (1850) and generally flank bridging channels (1660), the concentrated compression provided by lips (1854) against layers (1700, 1750) may minimize the risk of fluid leaking from PCR chambers (1450) and/or bridging channels (1660) during use of process chip (1400). Additional lips (1856) are provided on other regions of upper surface (1802); and also provide concentrated compression against layers (1700, 1750) when process chip (1400) is assembled. In some versions, each lip (1856) may be at least partially received by a corresponding notch (1754) of fifth layer (1750).
  • each PCR chamber portion (1850) includes a floor (1860), a sidewall (1870), and a shelf (1880) that is elevated relative to floor (1860).
  • a pair of sidewall channels (1872) are defined in each sidewall (1870).
  • Each pair of sidewall channels (1872) is positioned to correspond with the position of notches (1704), bridging channels (1660), and gaps (1855), such that each gap (1855) extends between corresponding adjacent sidewall channels (1872); such that each sidewall channel (1872) has an adjacent notch (1704); and such that notches (1704) are positioned between corresponding adjacent sidewall channels (1872).
  • FIGS. 38-40 show the combination of layers (1600, 1700, 1750, 1800), with layers (1500, 1900) omitted.
  • each sidewall channel (1872) extends from floor (1860) to notch (1704), with notch (1704) providing a path for fluid communication from sidewall channel (1872) to bridging channel (1660).
  • Another notch (1704) is positioned at the other end of bridging channel (1660) leading to another sidewall channel (1872) in another PCR chamber portion (1850).
  • channels (1660, 1872) and notches (1704) cooperate to provide a path for fluid communication from one PCR chamber (1450) to an adjacent PCR chamber (1450).
  • fluid may be driven from the “wet” region of one PCR chamber (1450) to the “wet” region of an adjacent PCR chamber (1450) via channels (1660, 1872) and notches (1704) in response to deformation of second layer (1900), in a manner similar to that described above in connection with FIGS. 22A-22B.
  • the fluid being communicated along bridging channels (1660) may be isolated from second layer (1900) by a thickness (T) of third layer (1600) that is defined between upper surface (1602) of third layer (1600) and the upper surface of each bridging channel (1660).
  • third layer (1600) that is between upper surface (1602) and bridging channels (1660) and that has thickness (T) may effectively shield second layer (1900) from the fluid being communicated along bridging channels (1660), thereby reducing or eliminating any potential risk of the fluid delaminating or otherwise damaging second layer (1900) as the fluid is communicated along bridging channels (1660).
  • Example 1 The fluidic apparatus of Example 1, the first layer comprising a non-metallic material.
  • Example 2 The fluidic apparatus of Example 2, the first layer comprising one or more of glass, plastic, or silicone.
  • the third chamber portion further defining a shelf within the fluid receiving volume.
  • Example 6 The fluidic apparatus of Example 6, the shelf of the third chamber portion defining a space to accommodate particles in the third chamber portion while fluid containing the particles is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the fourth chamber portion.
  • Example 9 The fluidic apparatus of any of Examples 1 through 7, further comprising a third layer interposed between the elastic layer and the second layer.
  • Example 8 The fluidic apparatus of Example 8, the third layer having an upper surface, the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion comprising a recess formed in the upper surface.
  • Example 9 The fluidic apparatus of Example 9, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 10 The fluidic apparatus of Example 10, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening.
  • Example 11 The fluidic apparatus of Example 11, the first sidewall including a first recess in fluid communication with the recess of the third chamber portion and the recess of the upper surface, the second sidewall including a second recess in fluid communication with the recess of the fourth chamber portion and the recess of the upper surface, such that the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion includes the first recess, the second recess, and the recess of the upper surface.
  • the second layer further including: (i) a first lip extending upwardly around an outer perimeter of the third chamber portion, and (ii) a second lip extending upwardly around an outer perimeter of the fourth chamber portion.
  • Example 14 [00215] The fluidic apparatus of Example 13, the first lip and the second lip to provide concentrated compression against at least one gasket layer disposed above the second layer.
  • Example 21 The fluidic apparatus of any of Examples 1 through 20, the third chamber portion having a depth of at least approximately 0 mm.
  • a fluidic apparatus comprising: (a) a first layer comprising a non-metallic material, the first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer comprising a metallic material, the second layer defining: (i) a third chamber portion defining a fluid receiving volume, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion defining a fluid receiving volume, the fourth chamber portion being positioned under the second chamber portion; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion.
  • the fluidic apparatus of Example 27 the sidewall of the third chamber portion defining a recess, the sidewall of the fourth chamber portion defining a recess, the fluidic apparatus further comprising a fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion, thereby providing a path for communication of fluid from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion.
  • Example 28 The fluidic apparatus of Example 28, further comprising a third layer interposed between the elastic layer and the second layer.
  • Example 30 The fluidic apparatus of Example 30, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 31 The fluidic apparatus of Example 31, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening.
  • Example 32 The fluidic apparatus of Example 32, the first sidewall including a first recess in fluid communication with the recess of the third chamber portion and the recess of the upper surface, the second sidewall including a second recess in fluid communication with the recess of the fourth chamber portion and the recess of the upper surface, such that the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion includes the first recess, the second recess, and the recess of the upper surface.
  • the fluidic apparatus of any of Examples 27 through 33 comprising one or more of glass, plastic, or silicone.
  • Example 36 The fluidic apparatus of Example 36, the shelf of the third chamber portion defining a space to accommodate particles in the third chamber portion while fluid containing the particles is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the fourth chamber portion.
  • the second layer further including: (i) a first lip extending upwardly around an outer perimeter of the third chamber portion, and (ii) a second lip extending upwardly around an outer perimeter of the fourth chamber portion.
  • Example 40 The fluidic apparatus of Example 38, the first lip and the second lip to provide concentrated compression against at least one gasket layer disposed above the second layer. [00266] Example 40
  • Example 47 The fluidic apparatus of any of Examples 27 through 45, the third chamber portion having a depth of at least approximately 0 mm.
  • a fluidic apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion defining a fluid receiving volume and a shelf within the fluid receiving volume, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion defining a fluid receiving volume and a shelf within the fluid receiving volume, the fourth chamber portion being positioned under the second chamber portion; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion, the shelf of the third chamber portion defining a space to accommodate particles in the third chamber portion while the fluid is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the fourth chamber portion.
  • Example 52 The fluidic apparatus of Example 52, the third chamber portion having a sidewall defining a recess, the fourth chamber portion having a sidewall defining a recess, the fluidic apparatus further comprising a fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion, thereby providing a path for communication of fluid from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion.
  • Example 53 The fluidic apparatus of Example 53, further comprising a third layer interposed between the elastic layer and the second layer.
  • Example 54 The fluidic apparatus of Example 54, the third layer having an upper surface, the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion comprising a recess formed in the upper surface.
  • Example 55 The fluidic apparatus of Example 55, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 56 The fluidic apparatus of Example 56, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening.
  • Example 57 The fluidic apparatus of Example 57, the first sidewall including a first recess in fluid communication with the recess of the third chamber portion and the recess of the upper surface, the second sidewall including a second recess in fluid communication with the recess of the fourth chamber portion and the recess of the upper surface, such that the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion includes the first recess, the second recess, and the recess of the upper surface.
  • Example 59 The fluidic apparatus of Example 59, the first layer comprising one or more of glass, plastic, or silicone.
  • Example 61 The fluidic apparatus of Example 61, the second layer comprising aluminum.
  • the second layer further including: (i) a first lip extending upwardly around an outer perimeter of the third chamber portion, and (ii) a second lip extending upwardly around an outer perimeter of the fourth chamber portion.
  • Example 63 The fluidic apparatus of Example 63, the first lip and the second lip to provide concentrated compression against at least one gasket layer disposed above the second layer.
  • Example 65 The fluidic apparatus of any of Examples 52 through 64, the fluid receiving volume of the third chamber portion providing a fluid capacity of at least approximately 0.3 mL.
  • Example 72 [00331] The fluidic apparatus of any of Examples 52 through 71, the third chamber portion having a depth of up to approximately 27.1 mm.
  • a method of using an apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion having a sidewall defining a recess and a fluid receiving volume adjacent to the recess, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion having a sidewall defining a recess and a fluid receiving volume adjacent to the recess, the fourth chamber portion being positioned under the second chamber portion; (c) a fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion, thereby providing a path for communication of fluid from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion; and (d) an elastic layer disposed
  • Example 77 The method of Example 77, further comprising performing polymerase chain reaction (PCR) amplification in one or both of the third chamber portion or the fourth chamber portion.
  • PCR polymerase chain reaction
  • Example 84 [00355] The method of Example 83, the shelf of the third chamber portion defining a space accommodating particles in the third chamber portion while the fluid is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the fourth chamber portion.
  • Example 84 The method of Example 84, the particles comprising beads.
  • a method of using an apparatus comprising: (a) a first layer comprising a non-metallic material, the first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer comprising a metallic material, the second layer defining: (i) a third chamber portion defining a fluid receiving volume, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion defining a fluid receiving volume, the fourth chamber portion being positioned under the second chamber portion; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion; the method comprising: (a) performing polymerase chain reaction (PCR) amplification in a fluid in the third chamber portion; and (b) deforming the elastic
  • Example 86 The method of Example 86, the fluid path extending from a recess in a sidewall of the third chamber portion to a recess in a sidewall of the fourth chamber portion, such that the fluid is driven from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion via the recess in the sidewall of the third chamber portion.
  • Example 88 Example 88
  • Example 91 The method of Example 91, the shelf of the third chamber portion defining a space accommodating particles in the third chamber portion while the fluid is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the fourth chamber portion.
  • Example 92 The method of Example 92, the particles comprising beads.
  • a method of using an apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion defining a fluid receiving volume and a shelf within the fluid receiving volume, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion defining a fluid receiving volume and a shelf within the fluid receiving volume, the fourth chamber portion being positioned under the second chamber portion; and (c) an elastic layer disposed between the first layer and the second layer; the method comprising deforming the elastic layer into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion, the shelf of the third chamber portion defining a space to accommodate particles in the third chamber portion while the fluid is driven from the fluid receiving volume of the third chamber portion into the fluid receiving volume of the
  • Example 94 The method of Example 94, the apparatus further including a fluid path extending from a recess in the sidewall of the third chamber portion to a recess in the sidewall of the fourth chamber portion, thereby providing a path for communication of fluid from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion.
  • Example 95 The method of Example 95, the fluid being driven from the fluid receiving volume of third chamber portion into the fluid receiving volume of fourth chamber portion via the fluid path.
  • Example 99 [00385] The method of any of Examples 94 through 98, further comprising applying heat to the second layer.
  • a fluidic apparatus comprising: (a) a first layer, the first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer comprising a metallic material, the second layer defining a second chamber portion defining a fluid receiving volume, the second chamber portion being positioned under the first chamber portion, the fluid receiving volume being configured to polymerase chain reaction (PCR) amplification; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the second chamber portion to thereby drive fluid from the fluid receiving volume of the second chamber portion.
  • PCR polymerase chain reaction
  • Example 103 The fluidic apparatus of Example 103, the first layer comprising a non- metallic material.
  • the fluidic apparatus of any of Examples 103 through 104 the first layer further defining a third chamber portion configured to receive pressurized gas; the second layer further defining a fourth chamber portion defining a fluid receiving volume, the fourth chamber portion being positioned under the third chamber portion.
  • Example 105 The fluidic apparatus of Example 105, the elastic layer being deformable into the fluid receiving volume of the second chamber portion to thereby drive fluid from the fluid receiving volume of the second chamber portion into the fluid receiving volume of the fourth chamber portion.
  • Example 106 The fluidic apparatus of Example 106, a sidewall of the second chamber portion defining a recess, the fluidic apparatus further comprising a fluid path extending from the recess in the sidewall of the second chamber portion to the fourth chamber portion, thereby providing a path for communication of fluid from the fluid receiving volume of the third chamber portion to the fluid receiving volume of the fourth chamber portion.
  • Example 107 The fluidic apparatus of Example 107, further comprising a third layer interposed between the elastic layer and the second layer.
  • Example 108 The fluidic apparatus of Example 108, the third layer having an upper surface, the fluid path extending from the recess in the sidewall of the second chamber portion to a recess in a sidewall of the fourth chamber portion further comprising a recess formed in the upper surface.
  • Example 109 The fluidic apparatus of Example 109, the third layer defining a first opening positioned over the second chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 111 [00409] The fluidic apparatus of Example 110, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening.
  • Example 111 The fluidic apparatus of Example 111, the first sidewall including a first recess in fluid communication with the recess of the second chamber portion and the recess of the upper surface, the second sidewall including a second recess in fluid communication with the recess of the fourth chamber portion and the recess of the upper surface, such that the fluid path extending from the recess in the sidewall of the second chamber portion to the recess in the sidewall of the fourth chamber portion includes the first recess, the second recess, and the recess of the upper surface.
  • the second layer further including: (i) a first lip extending upwardly around an outer perimeter of the second chamber portion, and (ii) a second lip extending upwardly around an outer perimeter of the fourth chamber portion.
  • Example 117 The fluidic apparatus of Example 117, the first lip and the second lip to provide concentrated compression against at least one gasket layer disposed above the second layer.
  • Example 124 The fluidic apparatus of any of Examples 103 through 118, the fluid receiving volume of the second chamber portion providing a fluid capacity of approximately 13.1 mL.
  • a method of using an apparatus comprising: (a) a first layer, the first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer comprising a metallic material, the second layer defining a second chamber portion defining a fluid receiving volume, the second chamber portion being positioned under the first chamber portion, the fluid receiving volume being configured to polymerase chain reaction (PCR) amplification; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the second chamber portion to thereby drive fluid from the fluid receiving volume of the second chamber portion; the method comprising: (a) performing polymerase chain reaction (PCR) amplification in a fluid in the second chamber portion; and (b) deforming the elastic layer into the fluid receiving volume of the second chamber portion to thereby drive the fluid from the second chamber portion.
  • PCR polymerase chain reaction
  • Example 131 The method of Example 131, the first layer further defining a third chamber portion configured to receive pressurized gas; the second layer further defining a fourth chamber portion defining a fluid receiving volume, the fourth chamber portion being positioned under the third chamber portion; the act of deforming the elastic layer into the fluid receiving volume of the second chamber portion to thereby drive the fluid from the second chamber portion resulting in at least some of the fluid being driven into the fluid receiving volume of the fourth chamber portion.
  • Example 135 The method of any of Examples 131 through 133, further comprising synthesizing a nucleic acid sequence template in the second chamber portion. [00456] Example 135
  • Example 137 The method of Example 137, the shelf of the second chamber portion defining a space accommodating particles in the second chamber portion while the fluid is driven from the fluid receiving volume of the second chamber portion.
  • Example 138 The method of Example 138, the particles comprising beads.
  • Example 8 The fluidic apparatus of Example 8, the third layer having a lower surface, the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion comprising a recess formed in the lower surface.
  • Example 140 The fluidic apparatus of Example 140, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 142 [00471] The fluidic apparatus of Example 141, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the recess extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the third and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the third and fourth chamber portions.
  • Example 143 The fluidic apparatus of Example 143, the first lip and the second lip to provide concentrated compression against at least one gasket layer interposed between the second and third layers.
  • Example 148 [00483] The fluidic apparatus of Example 147, the elastic layer being isolated from the bridging channel by a thickness of a third layer interposed between the elastic layer and the second layer.
  • Example 147 The fluidic apparatus of Example 147, a portion of the third layer being configured to shield the elastic layer from fluid within the bridging channel.
  • Example 150 The fluidic apparatus of Example 150, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 151 The fluidic apparatus of Example 151, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the recess extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the third and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the third and fourth chamber portions.
  • Example 155 The fluidic apparatus of Example 153, the first lip and the second lip to provide concentrated compression against at least one gasket layer interposed between the second and third layers.
  • Example 157 The fluidic apparatus of Example 157, the elastic layer being isolated from the bridging channel by a thickness of a third layer interposed between the elastic layer and the second layer.
  • Example 157 The fluidic apparatus of Example 157, a portion of the third layer being configured to shield the elastic layer from fluid within the bridging channel.
  • Example 54 The fluidic apparatus of Example 54, the third layer having a lower surface, the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion comprising a recess formed in the lower surface.
  • Example 161 The fluidic apparatus of Example 160, the third layer defining a first opening positioned over the third chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 161 The fluidic apparatus of Example 161, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the recess extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the third and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the third and fourth chamber portions.
  • Example 167 The fluidic apparatus of any of Examples 52 through 76 or 160 through 166, the fluid path extending from the recess in the sidewall of the third chamber portion to the recess in the sidewall of the fourth chamber portion comprising a bridging channel.
  • Example 167 The fluidic apparatus of Example 167, the elastic layer being isolated from the bridging channel by a thickness of a third layer interposed between the elastic layer and the second layer.
  • Example 167 The fluidic apparatus of Example 167, a portion of the third layer being configured to shield the elastic layer from fluid within the bridging channel.
  • Example 170 The method of Example 170, the apparatus further comprising a third layer interposed between the elastic layer and the second layer, the fluid being isolated from the elastic layer by a thickness of the third layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path.
  • Example 174 The method of Example 172, the apparatus further comprising a third layer interposed between the elastic layer and the second layer, the fluid being isolated from the elastic layer by a thickness of the third layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path.
  • Example 174 The method of Example 174, the apparatus further comprising a third layer interposed between the elastic layer and the second layer, the fluid being isolated from the elastic layer by a thickness of the third layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path.
  • Example 108 The fluidic apparatus of Example 108, the third layer having a lower surface, the fluid path extending from the recess in the sidewall of the second chamber portion to a recess in a sidewall of the fourth chamber portion further comprising a recess formed in the lower surface.
  • Example 176 The fluidic apparatus of Example 176, the third layer defining a first opening positioned over the second chamber portion and a second opening positioned over the fourth chamber portion.
  • Example 177 The fluidic apparatus of Example 177, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the recess extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the second and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the second and fourth chamber portions.
  • Example 180 [00547] The fluidic apparatus of Example 179, the first lip and the second lip to provide concentrated compression against at least one gasket layer interposed between the second and third layers.
  • Example 183 The fluidic apparatus of Example 183, the elastic layer being isolated from the bridging channel by a thickness of a third layer interposed between the elastic layer and the second layer.
  • Example 184 The fluidic apparatus of Example 184, a portion of the third layer being configured to shield the elastic layer from fluid within the bridging channel.
  • Example 187 The method of any of Examples 133 through 139, the fluid being isolated from the elastic layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path. [00560] Example 187
  • Example 186 The method of Example 186, the apparatus further comprising a third layer interposed between the elastic layer and the second layer, the fluid being isolated from the elastic layer by a thickness of the third layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path.
  • a fluidic apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion configured to receive fluid, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion configured to receive fluid, the fourth chamber portion being positioned under the second chamber portion; (c) a bridging channel extending between the third chamber portion and the fourth chamber portion; and (d) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the third chamber portion to thereby drive fluid into the fourth chamber portion via the bridging channel, the elastic layer being isolated from the bridging channel.
  • Example 188 The fluidic apparatus of Example 188, further comprising a third layer interposed between the elastic layer and the second layer.
  • Example 192 The fluidic apparatus of any of Examples 189 through 190, the third layer having a lower surface, the bridging channel comprising a recess formed in the lower surface. [00570]
  • Example 192 The fluidic apparatus of any of Examples 189 through 190, the third layer having a lower surface, the bridging channel comprising a recess formed in the lower surface.
  • Example 192 The fluidic apparatus of Example 192, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the bridging channel extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the third and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the third and fourth chamber portions.
  • Example 194 The fluidic apparatus of Example 194, the first lip and the second lip to provide concentrated compression against at least one gasket layer disposed above the second layer.
  • a fluidic apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion configured to receive fluid, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion configured to receive fluid, the fourth chamber portion being positioned under the second chamber portion; (c) a third layer having a lower surface, the third layer defining a recess in the lower surface for communicating fluid between the third and fourth chamber portions; and (d) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the third chamber portion to thereby drive fluid into the fourth chamber portion via the recess, the third layer being interposed between the elastic layer and the second layer.
  • Example 198 The fluidic apparatus of Example 198, the elastic layer being isolated from the recess by a thickness of the third layer.
  • Example 200 The fluidic apparatus of Example 200, the third layer further having a first sidewall defining the first opening and a second sidewall defining the second opening, the recess extending from the first sidewall to the second sidewall.
  • the second layer further including: (i) a first lip extending upwardly partially around outer perimeters of the third and fourth chamber portions, and (ii) a second lip extending upwardly partially around the outer perimeters of the third and fourth chamber portions.
  • Example 203 [00593] The fluidic apparatus of Example 202, the first lip and the second lip to provide concentrated compression against at least one gasket layer interposed between the second and third layers.
  • a method of using an apparatus comprising: (a) a first layer defining: (i) a first chamber portion, the first chamber portion being configured to receive pressurized gas, and (ii) a second chamber portion, the second chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a third chamber portion configured to receive fluid, the third chamber portion being positioned under the first chamber portion, and (ii) a fourth chamber portion configured to receive fluid, the fourth chamber portion being positioned under the second chamber portion; (c) a fluid path extending from the third chamber portion to the fourth chamber portion; and (d) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion; the method comprising deforming the elastic layer into the fluid receiving volume of the third chamber portion to thereby drive fluid into the fluid receiving volume of the fourth chamber portion via the fluid path, the fluid being isolated from the elastic layer
  • Example 207 [00601] The method of Example 206, the apparatus further comprising a third layer interposed between the elastic layer and the second layer, the fluid being isolated from the elastic layer by a thickness of the third layer as the fluid is driven into the fluid receiving volume of the fourth chamber portion via the fluid path.
  • Some versions of the examples described herein may be implemented using a processor, which may be part of a computer system and communicate with a number of peripheral devices via bus subsystem. Versions of the examples described herein that are implemented using a computer system may be implemented using a general- purpose computer that is programmed to perform the methods described herein. Alternatively, versions of the examples described herein that are implemented using a computer system may be implemented using a specific-purpose computer that is constructed with hardware arranged to perform the methods described herein. Versions of the examples described herein may also be implemented using a combination of at least one general-purpose computer and at least one specific-purpose computer.
  • each processor may include a central processing unit (CPU) of a computer system, a microprocessor, an application-specific integrated circuit (ASIC), other kinds of hardware components, and combinations thereof.
  • a computer system may include more than one type of processor.
  • the peripheral devices of a computer system may include a storage subsystem including, for example, memory devices and a file storage subsystem, user interface input devices, user interface output devices, and a network interface subsystem. The input and output devices may allow user interaction with the computer system.
  • the network interface subsystem may provide an interface to outside networks, including an interface to corresponding interface devices in other computer systems.
  • User interface input devices may include a keyboard; pointing devices such as a mouse, trackball, touchpad, or graphics tablet; a scanner; a touch screen incorporated into the display; audio input devices such as voice recognition systems and microphones; and other types of input devices.
  • pointing devices such as a mouse, trackball, touchpad, or graphics tablet
  • audio input devices such as voice recognition systems and microphones
  • input device is intended to include all possible types of devices and ways to input information into computer system.
  • a user interface output device may include a display subsystem, a printer, a fax machine, or non-visual displays such as audio output devices.
  • the display subsystem may include a cathode ray tube (CRT), a flat-panel device such as a liquid crystal display (LCD), a projection device, or some other mechanism for creating a visible image.
  • the display subsystem may also provide a non-visual display such as audio output devices.
  • output device is intended to include all possible types of devices and ways to output information from computer system to the user or to another machine or computer system.
  • a storage subsystem may store programming and data constructs that provide the functionality of some or all of the modules and methods described herein. These software modules may be generally executed by the processor of the computer system alone or in combination with other processors.
  • Memory used in the storage subsystem may include a number of memories including a main random-access memory (RAM) for storage of instructions and data during program execution and a read only memory (ROM) in which fixed instructions are stored.
  • RAM main random-access memory
  • ROM read only memory
  • a file storage subsystem may provide persistent storage for program and data files, and may include a hard disk drive, a floppy disk drive along with associated removable media, a CD-ROM drive, an optical drive, or removable media cartridges.
  • the modules implementing the functionality of certain implementations may be stored by file storage subsystem in the storage subsystem, or in other machines accessible by the processor.
  • the computer system itself may be of varying types including a personal computer, a portable computer, a workstation, a computer terminal, a network computer, a television, a mainframe, a server farm, a widely-distributed set of loosely networked computers, or any other data processing system or user device.
  • a personal computer a portable computer
  • a workstation a computer terminal
  • a network computer a television
  • mainframe a mainframe
  • server farm a widely-distributed set of loosely networked computers, or any other data processing system or user device.
  • the example of the computer system described herein is intended only as a specific example for purposes of illustrating the technology disclosed. Many other configurations of a computer system are possible having more or fewer components than the computer system described herein.
  • a non-transitory computer readable medium may be loaded with program instructions executable by a processor.
  • the program instructions when executed, implement one or more of the computer-implemented methods described above.
  • the program instructions may be loaded on a non-transitory CRM and, when combined with appropriate hardware, become a component of one or more of the computer- implemented systems that practice the methods disclosed.

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Abstract

Un appareil fluidique comprend une première couche définissant des première et deuxième parties de chambre, qui sont conçues pour recevoir un gaz sous pression. Une seconde couche définit des troisième et quatrième parties de chambre, qui ont chacune une paroi latérale respective définissant un évidement et un volume de réception de fluide adjacent à l'évidement. La troisième partie chambre est positionnée sous la première partie chambre. La quatrième partie chambre est positionnée sous la deuxième partie chambre. Un chemin fluidique s'étend de l'évidement dans la paroi latérale de la troisième partie de chambre à l'évidement dans la paroi latérale de la quatrième partie de chambre, fournissant un chemin pour la communication fluidique de la troisième chambre à la quatrième partie de chambre. Une couche élastique disposée entre la première couche et la seconde couche est déformable dans le volume de réception de fluide de la troisième partie de chambre pour ainsi entraîner le fluide dans le volume de réception de fluide de la quatrième partie de chambre.
PCT/US2024/055438 2023-11-13 2024-11-12 Ensemble puce microfluidique avec train roulant métallique Pending WO2025106391A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9895692B2 (en) * 2010-01-29 2018-02-20 Micronics, Inc. Sample-to-answer microfluidic cartridge
US10295441B2 (en) * 2013-04-26 2019-05-21 Robert Bosch Gmbh Method and device for producing a microfluidic analysis cartridge
US10406522B2 (en) * 2012-02-16 2019-09-10 Nri R&D Patent Licensing, Llc Three-dimensional multiple-layer microfluidic micro-droplet arrays for chemical and biochemical microreactors, miniature bioreactors, heat transfer, and other applications
US10710083B2 (en) * 2016-04-15 2020-07-14 University Of Maryland Integrated thermoplastic chip for rapid PCR and HRMA
US10946376B2 (en) * 2013-07-05 2021-03-16 Thinxxs Microtechnology Ag Carrier element for introducing a dry substance into a flow cell

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9895692B2 (en) * 2010-01-29 2018-02-20 Micronics, Inc. Sample-to-answer microfluidic cartridge
US10406522B2 (en) * 2012-02-16 2019-09-10 Nri R&D Patent Licensing, Llc Three-dimensional multiple-layer microfluidic micro-droplet arrays for chemical and biochemical microreactors, miniature bioreactors, heat transfer, and other applications
US10295441B2 (en) * 2013-04-26 2019-05-21 Robert Bosch Gmbh Method and device for producing a microfluidic analysis cartridge
US10946376B2 (en) * 2013-07-05 2021-03-16 Thinxxs Microtechnology Ag Carrier element for introducing a dry substance into a flow cell
US10710083B2 (en) * 2016-04-15 2020-07-14 University Of Maryland Integrated thermoplastic chip for rapid PCR and HRMA

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