Microfluidic apparatus with asymmetrical chambers

Abstract

A fluidic apparatus includes a first layer defining a first upper chamber portion. The first upper chamber portion is configured to receive pressurized gas. The first upper chamber portion has a first height. The fluid apparatus also includes a second layer defining a first lower chamber portion positioned under the first upper chamber portion. The first lower chamber portion is configured to receive at least one liquid. The first lower chamber has a second height different from the first height. The fluidic apparatus further includes an elastic layer disposed between the first layer and the second layer. The elastic layer is deformable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion. The elastic layer is configured to be substantially flat when the first lower chamber portion receives the at least one liquid.

Description

MICROFLUIDIC APPARATUS WITH ASYMMETRICAL CHAMBERSPRIORITY

[0001] This application claims the benefit of U.S. Pat. App. No. 63 / 681,918, entitled "Microfluidic Apparatus with Asymmetrical Chambers," filed August 12, 2024, the disclosure of which is incorporated by reference herein.BACKGROUND

[0002] The subject matter discussed in this section should not be assumed to be prior art merely as a result of its mention in this section. Similarly, a problem mentioned in this section or associated with the subject matter provided as background should not be assumed to have been previously recognized in the prior art. The subj ect matter in this section merely represents different approaches, which in and of themselves may also correspond to implementations of the claimed technology.

[0003] Some currently available technologies for manufacturing and formulating polynucleotide therapeutics (e.g., mRNA therapeutics, etc.) 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.BRIEF DESCRIPTION OF THE DRAWINGS

[0004] The details of one or more implementations are set forth in the accompanying drawings and the description below. Other features, aspects, and advantages will become apparent from the description, the drawings, and the claims, in which:

[0005] FIG. 1 depicts a schematic view of an example of a system including a microfluidic process chip;

[0006] FIG. 2 depicts an exploded perspective view of examples of components of the system of FIG. 1;

[0007] FIG. 3 depicts a top plan view of an example of a process chip that may be incorporated into the system of FIG. 1;

[0008] FIG. 4A depicts a cross-sectional side view of the process chip of FIG. 3 in a first state of operation;

[0009] FIG. 4B depicts a cross-sectional side view of the process chip of FIG. 3 in a second state of operation;

[0010] FIG. 4C depicts a cross-sectional side view of the process chip of FIG. 3 in a third state of operation;

[0011] FIG. 4D depicts a cross-sectional side view of the process chip of FIG. 3 in a fourth state of operation;

[0012] FIG. 4E depicts a cross-sectional side view of the process chip of FIG. 3 in a fifth state of operation;

[0013] FIG. 4F depicts a cross-sectional side view of the process chip of FIG. 3 in a sixth state of operation;

[0014] FIG. 5 depicts a top plan view of another example of a process chip that may be incorporated into the system of FIG. 1, with certain layers being transparent to show internal features;

[0015] FIG. 6 depicts an enlarged top plan view of area 6 of the process chip of FIG. 5 as indicated in FIG. 5;

[0016] FIG. 7 depicts a cross-sectional view of the process chip of FIG. 5, taken along line 7-7 in FIG. 5;

[0017] FIG. 8 depicts a cross-sectional view of another example of a process chip, showing various alternative configurations for a process chamber thereof;

[0018] FIG. 9 depicts a cross-sectional view of another example of a process chip, showing various alternative configurations for a process chamber thereof;

[0019] FIG. 10 depicts a top plan view of another example of a process chip that may be incorporated into the system of FIG. 1, with certain layers being transparent to show internal features;

[0020] FIG. 11 depicts an enlarged top plan view of area 11 of the process chip of FIG. 10 as indicated in FIG. 10;

[0021] FIG. 12 depicts a cross-sectional view of the process chip of FIG. 10, taken along line 12-12 in FIG. 10;

[0022] FIG. 13A depicts a cross-sectional view of the process chip of FIG. 10, taken along line 13-13 in FIG. 10, with a second layer of the process chip in a non-deformed state;

[0023] FIG. 13B depicts a cross-sectional view of the process chip of FIG. 10, taken along line 13- 13 in FIG. 10, with the second layer of the process chip in a first deformed state;

[0024] FIG. 13C depicts a cross-sectional view of the process chip of FIG. 10, taken along line 13-13 in FIG. 10, with the second layer of the process chip in a second deformed state; and

[0025] FIG. 14 depicts a cross-sectional view of the process chip of FIG. 10, taken along line 14-14 in FIG. 10.DET AILED DESCRIPTION

[0026] In some aspects, apparatuses and methods are disclosed herein for processing therapeutic polynucleotides. In particular, 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 anearly 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.

[0027] 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. In some scenarios, 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.).

[0028] I. Terminology

[0029] Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising” means various components may be co-jointly employed in the methods and articles (e.g., compositions and apparatuses including device and methods). For example, the term “comprising” will be understood to imply the inclusion of any stated elements or steps but not the exclusion of any other elements or steps. In general, 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.

[0030] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items and may be abbreviated as

[0031] 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. Similarly, the terms “upwardly,” “downwardly,” “vertical,” “horizontal,” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.

[0032] 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. 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. Although described or shown with respect to one embodiment, the features and elements so described or shown may apply to other embodiments. It will also be appreciated by those skilled in the art that references to a structure or feature that is disposed “adjacent” another feature may have portions that overlap or underlie the adjacent feature.

[0033] As used herein in the specification and claims, including as used in the examples and unless otherwise expressly specified, all numbers may be read as if prefaced by the word “about” or “approximately,” even if the term does not expressly appear. The phrase “about” or “approximately” may be used when describing magnitude and / or position to indicate that the value and / or position described is within a reasonable expected range of values and / or positions. For example, 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.

[0034] It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value,” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “X” is disclosed the “less than or equal to X” as well as “greater than or equal to X” (e.g., where X is a numerical value) is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11. 12, 13, and 14 are also disclosed.

[0035] Although the terms “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 belowcould 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.

[0036] As used herein, the terms '‘system,’7“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.

[0037] As used herein, “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. Aspects of this disclosure include 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 270, about 280, about 290, or about 300 nucleotides), or long polynucleotides having a length greater than about 300 nucleotides (e.g., polynucleotides of between about 300 to about 400 nucleotides, betw een about 400 to about 500 nucleotides, between about 500 to about 600 nucleotides, between about 600 to about 700 nucleotides, between about 700 to about 800 nucleotides, between about 800 to about 900 nucleotides, between about 900 to about 1000 nucleotides, between about 300 to about 500 nucleotides, between about 300 to about 600 nucleotides, between about 300 to about 700 nucleotides, between about 300 to about 800 nucleotides, between about 300 to about 900 nucleotides, or about 1000 nucleotides in length, or even greater than about 1000 nucleotides in length, such as about 1500, about 2000, about 2500, about 5000, or about 7000 nucleotides in length). Where a polynucleotide is double-stranded, its length may be similarly described in terms of base pairs.

[0038] As used herein "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.

[0039] As used herein a “cassette'’ (e.g., a synthetic in vitro transcription facilitator 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. In some aspects, 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.

[0040] As used herein, 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.

[0041] As used herein, the term “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.

[0042] As used herein, “m 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, ribozy mes, 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.

[0043] As used herein 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.

[0044] As used herein a “therapeutic polynucleotide composition” (or “therapeutic composition” for short) 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. An example of 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.

[0045] As used herein, “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.). Referring to something as a delivery vehicle need not exclude the possibility^ of the delivery vehicle also havingtherapeutic effects. Some versions of a delivery vehicle may provide additional therapeutic effects. In some versions, 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.”

[0046] As used herein, “joining” refers to methods such as ligation, synthesis, primer extension, annealing, recombination, or hybridization use to couple one component to another.

[0047] As used herein “purifying” refers to physical and / or chemical separation of a component (e.g., particles) of other unwanted components (e.g., contaminating substances, fragments, etc.).

[0048] As used herein, 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.

[0049] II. Overview of System Including Microfluidic Process Chip

[0050] 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). In other words, system (100) includes a chip-receiving component that is configured to removably accommodate a process chip (11 1), where the process chip (111) itself defines one or more microfluidic channels or fluid pathways. Components of system (100) (e.g., within housing (103)) that fluidically interact with process chip (111) 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)). In some versions, 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).

[0051] In some scenarios, 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). When process chip (1 1 1 ) 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.

[0052] 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.

[0053] In some variations, 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 (1 13) 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). In some variations, 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 (113).

[0054] As shown in FIG. 1, 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 (1 17) to one or more interior regions of process chip (111) as will be described in greater detail below . While only one pressure source (117) is shown, system (100) may include two or more pressure sources (117). In some scenarios, pressure may be generated by one or more sources other than pressure source (1 17). For instance, one or more vials or other fluid sources within reagent storage frame (107) may be pressurized. In addition, or in the alternative, reactions and / or other processes carried out on process chip (111) may generate additional fluid pressure. In the present example, 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 .

[0055] In some versions, pressurized fluid (e.g., gas) from at least one pressure source (117) reaches fluid interface assembly (109) via reagent storage frame (107), such that reagent storage frame (107) includes one or more components interposed in the fluid path between pressure source (1 17) and fluid interface assembly (109). In some versions, 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) bypassesreagent storage frame (107) to reach fluid interface assembly (109). Regardless of whether the fluid interface assembly (109) is interposed in the fluid path between pressure source (117) and 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. As described in greater detail below, pressure source (117) may selectively pressurize one or more chamber regions on process chip (111). In addition, or in the alternative, pressure source may also selectively pressurize one or more vials or other fluid storage containers held by reagent storage frame (107).

[0056] 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). In some versions, 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). In addition, or in the alternative, 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). In some such scenarios, the first process chip (11 1) 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.

[0057] 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 (1 11) when process chip (111) is held in seating mount (1 15). Any associated tubing (e.g., the fluid lines and / or the pressure lines) may be part of fluid interface assembly (109) and / or may connect to fluid interface assembly (109). In some versions, 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). In some versions, 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. In the present example, 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.

[0058] 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). In some versions, the vials may be pressurized by pressure source (117). Negative or positive pressure may thus be applied. For example, the fluid vials may be pressurized to between about 1 and about 20 psig (e.g., 5 psig, 10 psig, etc.). Alternatively, a vacuum (e.g., about -7 psig or about 7 psia) may be applied to draw fluids back into the vials (e.g., vials serving as storage depots) at the end of the process. The fluid vials may be driven at lower pressure than the pneumatic valves as described below, which may prevent or reduce leakage. In some variations, the difference in pressure between the fluid and pneumatic valves may be between about 1 psi and about 25 psi (e.g., about 3 psi, about 5 psi, 7 psi, 10 psi, 12 psi, 15 psi, 20 psi, etc.).

[0059] 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).

[0060] System (100) of the present example further includes one or more sensors (105). In some versions, such 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 movementwithin a process chip (111) that is mounted within seating mount (115). and / or other optically detectable conditions. In versions where 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). In some versions, 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. In some other versions, more than one sensor (105) is used to view such conditions. In some such versions, 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.

[0061] In versions where sensors (105) include at least one optical sensor, visual / optical markers may be used to estimate yield. For example, fluorescence may be used to detect process yield or residual material by tagging with fluorophores. In addition, or in the alternative, dynamic light scattering (DLS) may be used to measure particle size distributions within a portion of the process chip (111) (e.g., such as a mixing portion of process chip (1 1 1)). In some variations, 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). In versions where 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.

[0062] In versions where sensors (105) include at least one optical sensor that is configured to capture video images, such sensors (105) may record at least some activity on process chip (111). For example, an entire run for synthesizing and / or processing a material (e.g., a therapeutic RNA) may be recorded by one or more video sensors (105), including a video sensor (105) that may visualize process chip (111) (e.g., from above). Processing on process chip (111) may be visually tracked and this video record may be retained for later quality control and / or processing. Thus, thevideo record of the processing may be saved, stored, and / or transmitted for subsequent review and / or analysis. In addition, as will be described in greater detail below, 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.

[0063] System (100) of the present example may be controlled by a controller (121). Controller (121) may include one or more processors, one or more memories, and various other suitable electrical components. In some versions, one or more components of controller (121) (e.g., one or more processors, etc.) is / are embedded within system (100) (e.g., contained within housing (103)). In addition, or in the alternative, one or more components of controller (121) (e.g., one or more processors, etc.) may be detachably attached or detachably connected with other components of system (100). Thus, at least a portion of controller (121) may be removable. Moreover, at least a portion of controller (121) may be remote from housing (103) in some versions.

[0064] The control by 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). In some versions, controller (121) is fully automated to a point where user inputs are not needed. In some such versions, 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.

[0065] FIG. 2 shows examples of certain forms that may be taken by various components of system (100). In particular, 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. These components are secured relative to a base (180). A set of rods (182) support reagent storage frame (150) over fluid interface assembly (152).

[0066] As shown in FIG. 2, 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). In addition, process chip (200) is w ithin 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).

[0067] While 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). For instance, 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. In some such versions, 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. While not shown, 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).

[0068] In some versions, 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).

[0069] 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). From the protocol, 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 (1 15); 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.After confirming the starting materials and equipment, 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).

[0070] III. Example of Process Chip

[0071] FIGS. 3 and 4A-4F depict the example of a process chip (200) in further detail. In combination with the rest of system (100), 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. As shown in FIG. 3, 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). As described in greater detail below, each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109). In the present example, 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.

[0072] As also shown in FIG. 3, 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. By way of example only, such 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). In some versions, fluidic connector (232) is operable like a valve between an open and closed state (e.g., similar to valve chamber (224)). In some other versions, fluidic connector (232) remains open throughout the process of makingthe therapeutic composition. In the present example, chambers (230) are used to provide synthesis of polynucleotides, though chambers (230) may alternatively sen e any other suitable purpose(s).

[0073] In the example shown in FIG. 3, 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. In versions where a chamber (250) is used for purification, chamber (250) may include a material that is configured to absorb selected moieties from a fluidic mixture in chamber (250). In some such versions, the material may include a cellulose material, which may selectively absorb double-stranded mRNA from a mixture. In some such versions, 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. Alternatively, chambers (250) may be used for any other suitable purpose.

[0074] Additional 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.

[0075] As shown in FIG. 3, 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. In some versions, fluid ports (221) are used to communicate additional fluids to chambers (270). In addition, or in the alternative, 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.

[0076] Process chip (200) further includes several reservoir chambers (260). In this example, 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 betw een reservoir chamber (260) and a corresponding fluid port (266). In some versions, each fluid port (266) is configured to communicate fluid from a corresponding vial in reagent storage frame (107) to a corresponding reservoir chamber (260). In addition, or in the alternative, 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). In the present example, reservoir chambers (260) are used to provide metering of fluid communicated to and / or from process chip (200). Alternatively, 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).

[0077] As also shown in FIG. 3, 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 correspondingpressure channel (244). As described in greater detail below, each pressure port (240) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109). In the present example, 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.

[0078] 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). In the example shown in FIG. 3, 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.

[0079] As shown in FIGS. 4A-4F, process chip (200) further includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306). As described in greater detail below, some versions of elastic layer (302) are in the form of a flexible membrane. First plate (300) has an upper surface (210) and a low er surface (310), with lower surface (310) apposing elastic layer (302). Second plate (304) has an upper surface (312) and alower 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). In the present example, another elastic layer (316) is also interposed between second and third plates (304, 306), though this elastic layer (316) is optional.

[0080] Plates (300, 304, 306) of the present example are substantially translucent to visible light and / or ultraviolet light. By “substantially translucent” is meant that at least 50% (e.g., at least 90%, including in some instances 100%) of light is transmitted through the material compared to a translucent material. In some variations, the one ormore of plates (300, 304, 306) may comprise materials that are substantially transparent to visible light and / or ultraviolet light. By "‘substantially transparent7’ is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a completely transparent material. As another example, one or more of plates (300, 304, 306) 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%.

[0081] 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).

[0082] Elastic layer (302) of the present example is formed as a liquid-impermeable flexible membrane. In some versions, elastic layer (302) is gas-permeable despite being liquid-impermeable. In some such versions, certain regions of elastic layer (302) are treated to be gas-permeable while the non-treated regions of elastic layer (302) are gas- impermeable. As described below, elastic layer (302) may be used to drive fluids across process chip (200) via peristaltic pumping action. As also described below, elastic layer (302) may be used to provide valves at various locations along process chip (200). In some versions, a single sheet of elastic material spans across the width of process chip (200) to form elastic layer (302). In some other versions, two or more discrete pieces of elastic material are used to form elastic layer (302), with such discrete pieces of elastic material being positioned at different locations across the width of process chip (200). By way of example only, elastic layer (302) may include a membrane comprising polydimethylsilicone (PDMS) elastomer fdm.

[0083] As best seen in FIGS. 4A-4F, 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. For instance, chamber (320) may be analogous to chamber (264), chamber (322) may be analogous to chamber (260), chamber (324) may be analogous to chamber (262), and chamber (326) may be analogous to chamber (250).

[0084] As shown in FIGS. 4A-4F, 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). As noted above, 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). In some versions, 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.

[0085] Pressure port (240) is formed through first plate (300). A corresponding opening (344) is formed through the region of elastic layer (302) underlying pressureport (240). Pressure channel (244) extends from opening (344) to upper chamber region (330) of first chamber (320). As noted above, 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). In some versions, 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).

[0086] While 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). In some versions, one or more of chambers (320. 322, 324. 326) has its own dedicated pressure line (208) and corresponding pressure channel (244). In addition, or in the alternative, 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. 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).

[0087] IV. Example of Valving and Peristaltic Pumping Driven via Elastic Layer

[0088] As noted above, 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. An example of such operation is illustrated in the sequence depicted through FIGS. 4A-4F. In this example,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.

[0089] 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). In 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). In this state, 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). This seating of elastic layer (302) against the surface of lower chamber region (332) of chamber (324) prevents fluid (207) from entering chamber (324), such that chamber (324) is operating like a closed valve in the state shown in FIG. 4B. The negatively pressurized gas in upper chamber regions (330) of chambers (320, 322) causes the corresponding portion of elastic layer (302) in chambers (320, 322) to deform and seat against upper chamber regions (330) of chambers (320, 322). This allows fluid (207) to occupy the full capacity of chambers (320, 322).

[0090] After reaching the state shown in FIG. 4B, positively pressurized gas is communicated to upper chamber region (330) of chamber (320) while the pneumatic state of chambers (322, 324) may remain unchanged. This results in the state shown in FIG. 4C. As shown, 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 show n in FIG. 4C. However, 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 offluid (207), such that only a precise, predetermined volume of fluid (207) is communicated further along process chip (200). By way of example only, such 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.

[0091] Once the appropriate metering volume has been achieved, negatively pressurized gas is communicated to upper chamber regions (330) of chambers (324, 326) while the pneumatic state of chambers (320, 322) may remain unchanged. This results in the state shown in FIG. 4D. As shown, the negatively pressurized gas in upper chamber regions (330) of chambers (324, 326) causes the corresponding portion of elastic layer (302) in chamber (324, 326) to deform and seat against the surface of upper chamber regions (330) of chambers (324, 326). This effectively opens the valve formed by chamber (324) and puts chamber (326) in a state to receive fluid (207). This also produces a negative pressure in chamber (324) that draws fluid (207) from chamber (322) into chamber (324).

[0092] With the valve formed by chamber (324) being in the open state, positively pressurized gas is communicated to upper chamber region (330) of chamber (322) while the pneumatic state of chambers (320, 324, 326) may remain unchanged. This results in the state shown in FIG. 4E. As shown, the positively pressurized gas in upper chamber region (330) of chamber (322) causes the corresponding portion of elastic layer (302) in chamber (322) to deform and seat against the surface of lower chamber region (332) of chamber (322). This deformation of elastic layer (302) drives fluid (207) out of chamber (322). Since the valve formed by chamber (320) is in a closed state and the valve formed by chamber (324) is in an open state, fluid (207) travels from chamber (322) into chamber (324). In the present example, the capacity of chamber (322) is greater than the capacity of chamber (324), such that fluid (207) from chamber (322) overflows from chamber (324) into chamber (326).

[0093] Once fluid (207) has been communicated from chamber (322) to chambers (324, 326), positively pressurized gas is communicated to upper chamber region (330) of chamber (324) while the pneumatic state of chambers (320. 322, 326) may remain unchanged. This results in the state shown in FIG. 4F. As shown, the positivelypressurized gas in upper chamber region (330) of chamber (324) causes the corresponding portion of elastic layer (302) in chamber (324) to deform and seat against the surface of lower chamber region (332) of chamber (324). This deformation of elastic layer (302) drives fluid (207) out of 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).

[0094] At the stage shown in FIG. 4F, 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. In addition, or in the alternative, 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. Regardless of what is done with fluid (207) after fluid (207) has reached chamber (326), it should be understood that 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. Such 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.”

[0095] In some scenarios, it may be desirable to remove air or other gas from one or more fluid pathways in process chip (200). To accomplish this, 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. For instance, such ventilation or evacuation may be performed as part of a priming process as fluid is initially introduced to process chip (200). In addition, or in the alternative, such ventilation or evacuation may be performed to relieve gas that is generated in the fluid during the process of forming the therapeutic composition. Such ventilation or gasrelief chambers may be referred to as “vacuum caps.” In some versions, at least the region of elastic layer (302) that is positioned in the vacuum cap (if not the entirety of elastic layer (302)) 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). In some versions, 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.

[0096] V. Example of Process Chip with Symmetrical Chambers

[0097] FIGS. 5-7 show an example of a process chip (400) that is similar to process chip (200) described above, except as otherwise described below. In this regard, in combination with the rest of system ( 100), process chip (400) may be utilized to provide in-vitro synthesis, purification, concentration, formulation (e.g., encapsulation of a therapeutic composition in a delivery vehicle through a mixing process, etc.), and / or analysis of therapeutic compositions and / or templates, including but not limited to therapeutic polynucleotides templates, therapeutic polynucleotides, and / or therapeutic polynucleotide compositions. Alternatively, process chip (400) may be utilized in other processes.

[0098] As shown in FIG. 5, 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 assembly7(109).

[0099] As shown in FIG. 5, process chip (400) of this example further includes a plurality7of process chambers (450, 460, 470) that may be used to serve different purposes during the process of producing the therapeutic composition as described herein. By way of example only, process chambers (450, 460, 470) may be used toprovide synthesis, purification, dialysis, compounding, and / or concentration of one or more therapeutic compositions; or to perform any other suitable function(s). In the present example, process chambers (450) are configured to be used for IVT reaction, and thus may also be referred to as “reaction chambers;” process chambers (460) are configured to be used for DNA digestion, and thus may also be referred to as “digestion chambers;” and process chambers (470) are configured to be used for RNA purification, and thus may also be referred to as “purification chambers.” Process chip (400) may have any other suitable number and / or type(s) of process chambers (450, 460, 470). As shown, process chip (400) of this example further includes a first layer (500), a second layer (600), a third layer (700), a fourth layer (800), and a fifth layer (900). Each of these layers (500, 600. 700, 800. 900). and their structural and functional relationships with each other, will be described in greater detail below.

[0100] First layer (500) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, first layer (500) may be substantially translucent to visible light and / or ultraviolet light. By “substantially translucent” is meant that at least 50% (e.g., 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). In some versions, first layer (500) is formed as a lamination of tw o or more layers of material, such that first layer (500) does not necessarily need to be formed as a single homogenous continuum of material.

[0101] As best shown in FIG. 7, first layer (500) includes an upper surface (502) and a lower surface (504). First layer (500) may include openings formed through both surfaces (502, 504), and such openings may form part of fluid ports (410) and / or pressure ports (420). A plurality of process chamber portions (550, 560, 570) are formed as recesses in lower surface (504), collectively providing a protruding region (552) in upper surface (502). Each process chamber portion (550. 560, 570) forms a “dry” region of a respective process chamber (450, 460, 470) as described herein.

[0102] Each process chamber portion (550. 560, 570) has a sidewall (553, 563, 573) that extends upwardly from lower surface (504) toward a corresponding ceiling (555,565, 575). As seen in FIG. 5, 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 forming a respective pressure port (420). Each pneumatic channel (522) is further configured to define a space between first layer (500) and second layer (600), such that pressurized gas may be communicated along pneumatic channels (522). Each process chamber portion (550, 560, 570) has its own associated pneumatic channel (522) and pressure port (420) in this example. Thus, each process chamber portion (550, 560, 570) may be pneumatically pressurized independently of the other process chamber portions (550, 560, 570).

[0103] Second layer (600) is interposed between first layer (500) and third layer (700). Second layer (600) may be in the form of first and second liquid-impermeable flexible membranes laminated to each other in a vertically stacked arrangement. As described below, second layer (600) may be used to drive fluids across process chip (400) via peristaltic pumping action. As also described below, second layer (600) may be used to provide valves at various locations along process chip (400). By way of example only, one or both flexible membranes of second layer (600) may comprise polydimethylsilicone (PDMS) elastomer film. Alternatively, any other suitable material(s) may be used to form second layer (600).

[0104] Second layer (600) may be configured and operable like elastic layer (302) described above. Second layer (600) may thus pass through chambers collectively defined by first and third layers (500, 700) to separate such chambers into an upper chamber region (which receives pneumatic pressure) and a lower chamber region (which receives fluid). Such chambers include process chambers (450, 460, 470), valve chambers as described herein, and pump chambers as described herein. Second layer (600) may further thus be pneumatically deformed to provide valving, peristaltic pumping, etc. within process chip (400). Second layer (600) may include a plurality of openings formed through the flexible membranes. The openings may be positioned at fluid ports (410) and / or pressure ports (420) to allow fluid and / or pressurized gas to be communicated through second layer (600) at fluid ports (410) and / or pressure ports (420), respectively. The distal ends of fluid and pressure lines (206, 208) may sealagainst corresponding regions of second layer (600) at or near the bottom ends of fluid and pressure ports (410, 420), respectively.

[0105] In some instances, second layer (600) may have a thickness of about 100 pm. By way of further example only, second layer (600) may be configured and operable in accordance with at least some of the teachings of International Pat. App. No. PCT / US2024 / 020488, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,’' filed on March 19, 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. For example, second layer (600) may include first and second liquid-impermeable flexible membranes (not shown) laminated to each other in a vertically stacked arrangement, and the second flexible membrane may include a plurality of apertures (e.g., cutouts) that are each configured to vertically align with a respective chamber (450, 460, 470), while the first flexible membrane may lack such apertures. In some such cases, the first flexible membrane may have a thickness of about 100 pm.

[0106] In the present example, the region of each chamber in process chip (400) that is above second layer (600) constitutes a “dry” chamber region since this region receives pressurized gas to pneumatically deflect second layer (600). Conversely, the region of each chamber in process chip (400) that is below second layer (600) constitutes a “wet” chamber region since this region receives fluid. However, these “wet” and “dry” roles may be reversed in some variations. In other words, some variations of process chip (400) may provide pressurized gas to the 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.

[0107] Third layer (700) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, third layer (700) is substantially translucent to visible light and / or ultraviolet light. Third layer (700) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, third layer (700) is formed as alamination of two or more layers of material, such that third layer (700) does not necessarily need to be formed as a single homogenous continuum of material.

[0108] As best shown in FIG. 7, third layer (700) includes an upper surface (702) and a lower surface (704). Third layer (700) may include openings formed through both surfaces (702, 704), and such openings may form part of fluid ports (410) and / or pressure ports (420). A plurality' of process chamber portions (750, 760, 770) are formed as recesses in upper surface (702), collectively providing a protruding region (752) in lower surface (704). Each process chamber portion (750. 760, 770) forms a “wet” region of a respective process chamber (450, 460, 470) as described herein.

[0109] Each process chamber portion (750. 760, 770) has a sidewall (753, 763, 773) that extends downwardly from upper surface (702) toward a corresponding floor (755, 765, 775). As seen in FIG. 5, upper surface (702) of third layer (700) further defines a plurality of fluid channels (712). which are formed as recesses in upper surface (702). Each fluid channel (712) is in fluid communication with a corresponding opening forming a respective fluid port (410). Each fluid channel (712) is further configured to define a space between third layer (700) and second layer (600), such that fluid may be communicated along fluid channels (712). Each process chamber portion (750, 760, 770) also has at least one respective fluid channel (712) leading thereto, such that fluid may be communicated to / from each process chamber (450, 460, 470) via the fluid channel (712) leading to the process chamber portion (750, 760, 770) associated with the process chamber (450, 460, 470). Corresponding pumps and valves interposed between an opening forming a respective fluid port (410) and each fluid channel (712) leading to each respective process chamber portion (750. 760, 770) may drive and regulate the communication of fluid to / from each process chamber (450, 460, 470).

[0110] As also seen in FIGS. 5-7, third layer (700) includes a plurality of bridging channels (751), sidewall channels (754), and floor channels (756). Sidewall channels (754) may also be referred to as “sippers.” Each bridging channel (751) is formed as a recess in upper surface (702); each floor channel (756) is formed as a recess in a corresponding floor (755); and each sidewall channel (754) is formed as a recess in a corresponding sidewall (753) that extends upwardly from the corresponding floorchannel (756) toward upper surface (702). The presence of bridging channels (751), sidewall channels (754). and / or floor channels (756) may provide a pathway for fluid to escape the respective process chamber (450, 460, 470) when second layer (600) is deformed downwardly toward the respective floor (755), such that sidewall channels (754) and / or floor channels (756) may prevent pockets of fluid from being trapped between localized regions of deformed second layer (600) and the respective floor (755) and / or sidewall (753) (or otherwise reduce the risk of such pockets being formed).

[0111] As shown, at least some sidewall channels (754) extend from the corresponding floor channel (756) to a corresponding bridging channel (751), such that each bridging channel (751) is flanked by a pair of adjacent sidewall channels (754). Each bridging channel (751) and the pair of adjacent sidewall channels (754) are configured to provide a path for communication of fluid from one process chamber (450, 460, 470) to an adjacent process chamber (450, 460, 470).

[0112] Referring to FIG. 7, fourth layer (800) may include an elastomeric membrane, such as PDMS or other silicone compositions, and / or any other suitable material(s). Fourth layer (800) may include a central opening that is sized and configured to receive protruding region (752) of third layer (700). Fourth layer (800) may provide a gasket forming a seal between adjacent regions of third layer (700) and another layer such as a rigid plate. Fifth layer (900) is sized and configured to directly underlie protruding region (752) of third layer (700), and is in the form of a thermal pad in this example. It will be appreciated that any suitable structures or techniques may be used to secure layers (500, 600. 700, 800, 900) together.

[0113] In the example shown, each upper process chamber portion (550, 560. 570) is sized and shaped substantially similarly to the corresponding lower process chamber portion (750, 760, 770), such that each process chamber (450, 460, 470) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (600) (e.g., notwithstanding the presence of sidewall channels (754), floor channels (756), and bridging channels (751) in third layer (700)). For example, with specific regard to reaction chambers (450), each upper process chamber portion (550) has an upper process chamber portion length (Lup), an upper process chamber portion maximumwidth (Wup,max), an upper process chamber portion minimum width (Wup,mm), and an upper process chamber portion height (HUP) defined between lower surface (504) and the respective ceiling (555); and each lower process chamber portion (750) has a lower process chamber portion length (LLO) substantially equal to the upper process chamber portion length (Lup), a lower process chamber portion maximum width (Wuo,max) substantially equal to the upper process chamber portion maximum width (Wup,max), a lower process chamber portion minimum width (Wi.am.n) substantially equal to the upper process chamber portion minimum width (Wup,min), and a lower process chamber portion height (HLO) defined between upper surface (702) and the respective floor (755) and substantially equal to the upper process chamber portion height (HUP). AS a result, each upper process chamber portion (550. 560, 570) may be configured to hold a first volume of fluid and each lower process chamber portion (750, 760, 770) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for the corresponding upper process chamber portion (550, 560, 570), such that each process chamber portion (550. 560, 570, 750, 760, 770) of a particular process chamber (450, 460, 470) may have an individual capacity substantially equal to about 50% of the full capacity of the respective process chamber (450, 460, 470).

[0114] With specific regard to reaction chambers (450), each upper process chamber portion (550) may provide the portion of second layer (600) that extends below upper process chamber portion (550) and that is configured to be deflected upwardly into upper process chamber portion (550) with a predefined stretch ratio within upper process chamber portion (550). In this regard, the stretch ratio within upper process chamber portion (550) may be defined as the ratio of the surface area of the portion of second layer (600) deflected fully upwardly into upper process chamber portion (550) vs. the surface area of the same portion undeflected (e g., horizontal); and may be expressed as a function of the upper process chamber portion height (HUP), the inverse of the upper process chamber portion maximum width (Wup.max), and the inverse of the upper process chamber portion length (LUP). For example, the stretch ratio may be estimated via the following equation: R = 1 + 2(HUP / LUP) + 2(Hup / Wup,max).

[0115] Likewise, each lower process chamber portion (750) may provide the portion of second layer (600) that extends above lower process chamber portion (750) and thatis configured to be deflected downwardly into lower process chamber portion (750) with a predefined stretch ratio within lower process chamber portion (750). In this regard, the stretch ratio within lower process chamber portion (750) may be defined as the ratio of the surface area of the portion of second layer (600) deflected fully downwardly into lower process chamber portion (750) vs. the surface area of the same portion undeflected (e.g., horizontal); and may be expressed as a function of the lower process chamber portion height (HLO), the inverse of the lower process chamber portion maximum width (WLo,max), and the inverse of the lower process chamber portion length (LLO). For example, the stretch ratio may be estimated via the following equation: R = 1 + 2(HLO / LLO) + 2(HLO / WLo,max).

[0116] The substantial symmetry of reaction chambers (450) may provide second layer (600) with a stretch ratio within upper process chamber portion (550) that is substantially the same as the stretch ratio within lower process chamber portion (750). For example the stretch ratio of second layer (600) within both upper and lower process chamber portions (750) may be about 500%.

[0117] As noted above, each bridging channel (751) may provide a path for communication of fluid from one process chamber (450, 460, 470) to an adjacent process chamber (450, 460, 470). For example, with specific regard to reaction chambers (450) shown in FIG. 6, fluid may be transferred back and forth between the “wet” regions of reaction chambers (450) via the corresponding bridging channels (751) to agitate a fluidic mixture. Such an agitation process, also referred to as a blending cycle, may initiate when the “wet” region of each reaction chamber (450) is filled with an initial amount of fluid (e.g., liquid) that exceeds the individual capacity of the corresponding lower process chamber portion (750). For example, the “wet” region of each reaction chamber (450) may be filled with an initial amount of fluid that is substantially equal to about 75% of the full capacity of the respective reaction chamber (450). Second layer (600) may be deflected partially upwardly into each of the corresponding upper process chamber portions (550) toward the respective ceiling (555) to accommodate the initial amount of fluid in the “w et” region of each reaction chamber (450), such as in instances where the fluid is substantially incompressible (e.g., where the fluid has a high water content).

[0118] With the “wet” region of each reaction chamber (450) filled with the initial amount of fluid, second layer (600) may be pneumatically deflected downwardly in a first one of the reaction chambers (450) (e.g., the lefthand reaction chamber (450) in FIG. 6) toward the respective floor (755) and upwardly in a second one of the reaction chambers (450) (e.g., the righthand reaction chamber (450) in FIG. 6) toward the respective ceiling (555), to thereby drive a portion of the fluid from the first reaction chamber (450) into the second reaction chamber (450). Second layer (600) may then be pneumatically deflected downwardly in the second reaction chamber (450) toward the respective floor (755) and upwardly in the first reaction chamber (450) toward the respective ceiling (555), to thereby drive a portion of the fluid from the second reaction chamber (450) into the first reaction chamber (450). Second layer (600) may alternate between being pneumatically deflected downwardly in the first reaction chamber (450) and upwardly in the second reaction chamber (450), and being pneumatically deflected downwardly in the second reaction chamber (450) and upwardly in the first reaction chamber (450), to thereby drive fluid back and forth between the first and second reaction chambers (450) to achieve the desired blending.

[0119] It will be appreciated that, when second layer (600) is pneumatically deflected upwardly in either reaction chamber (450) to drive fluid between the reaction chambers (450), second layer (600) may be deflected farther upwardly into the corresponding upper process chamber portion (550) and closer to the respective ceiling (555) than when second layer (600) is deflected partially upwardly into the corresponding upper process chamber portion (550) to accommodate the initial amount of fluid in the “wet” region of each reaction chamber (450). In some cases when second layer (600) is pneumatically deflected upwardly in either reaction chamber (450) to drive fluid between the reaction chambers (450), second layer (600) may be deflected fully upwardly into the corresponding upper process chamber portion (550) such that second layer (600) contacts the respective ceiling (555).

[0120] VI. Examples of Process Chips with Membrane Yield Mitigation

[0121] In some scenarios, it may be desirable to provide a version of process chip (400) that inhibits ripping or tearing of second layer (600) or otherwise reduces the riskof ripping or tearing of second layer (600) and thereby improve longevity of second layer (600). For instance, it may be desirable to reduce any tensioning (e.g.. stretching) of second layer (600) that may occur during a blending cycle, which may otherwise eventually cause second layer (600) to yield (e.g., after numerous blending cycles). It may therefore be desirable to provide a variation of process chip (400) with process chambers that are sized and configured to facilitate blending of a fluid mixture while minimizing tensioning (e.g., stretching) of second layer (600).

[0122] A. Examples of Alternative Types of Process Chambers

[0123] FIGS. 8-9 show examples of process chips (400a, 400b) with various alternative types of process chambers that may provide at least some, if not all, of the features and functionalities described above. As shown in FIGS. 8-9, process chips (400a, 400b) each include respective process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) that may be used to serve different purposes during the process of producing the therapeutic composition as described herein. By way of example only, process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) 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). In the present example, process chambers (450a, 450b, 450c, 450d, 450e. 450f, 450g, 450h) are each configured to be used for IVT reaction, and thus may also be referred to as “reaction chambers." Reaction chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) may each be similar to reaction chambers (450) described above, except as otherwise described below. As shown, process chips (400a, 400b) each further include a respective first layer (500a. 500b). a respective second layer (600a, 600b), and a respective third layer (700a, 700b), which may each be similar to first layer (500), second layer (600), and third layer (700) described above, respectively, except as otherwise described below.

[0124] As shown in FIGS. 8-9, each first layer (500a, 500b) includes an upper surface (502a, 502b) and a lower surface (504a, 504b). A plurality of process chamber portions (550a. 550b, 550c, 550d. 550e, 550f, 550g, 550h) are formed as recesses in each lower surface (504a, 504b), collectively providing a protruding region (552a,552b) in the respective upper surface (502a, 502b). Each process chamber portion (550a, 550b, 550c, 550d. 550e, 550f, 550g, 550h) forms a '’dry” region of a respective process chamber (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) as described herein. Each process chamber portion (550a, 550b, 550c, 550d, 550e, 550f, 550g, 550h) has a sidewall (553a, 553b, 553c, 553d, 553e, 553f, 553g, 553h) that extends upwardly from the respective lower surface (504a, 504b) toward a corresponding ceiling (555a, 555b, 555c, 555d, 555e, 555f, 555g, 555h). Each first layer (500a, 500b) further includes a plurality of pneumatic channels (522a, 522b).

[0125] As shown in FIGS. 8-9, each third layer (700a, 700b) includes an upper surface (702a, 702b) and a lower surface (704a, 704b). A plurality of process chamber portions (750a, 750b, 750c, 750d, 750e, 750f, 750g, 750h) are formed as recesses in each upper surface (702a. 702b). collectively providing a protruding region (752a, 752b) in the respective lower surface (704a, 704b). Each process chamber portion (750a, 750b, 750c, 750d, 750e, 750f, 750g, 750h) forms a “wet” region of a respective process chamber (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) as described herein. Each process chamber portion (750a, 750b, 750c, 750d. 750e, 750f, 750g, 750h) has a sidewall (753a, 753b753c, 753d, 753e. 753f. 753g, 753h) that extends downwardly from the respective upper surface (702a, 702b) toward a corresponding floor (755a, 755b, 755c, 755d, 755e, 755f, 755g, 755h). Each third layer (700a, 700b) further includes a plurality of fluid channels, a plurality of bridging channels (751a, 751b), and a plurality of sidewall channels (754a, 754b).

[0126] While process chambers (450a, 450b, 450c, 450d, 450e. 450f, 450g, 450h) are shown incorporated into process chips (400a, 400b) in FIGS. 8-9. any one or more of process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) may be incorporated into process chip (400) in place of any one or more of process chambers (450, 460, 470), such as in place of any one or more of reaction chambers (450). Thus, process chambers (450a, 450b. 450c, 450d, 450e, 450f, 450g, 450h) shown in FIGS. 8- 9 may be referred to as alternative process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) and / or as alternative reaction chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h). Each alternative process chamber (450a, 450b, 450c, 450d, 450e. 450f. 450g, 450h) is configured to provide the corresponding second layer (600a,600b) with a reduced tendency to yield, at least relative to that provided by process chamber (450) to second layer (600), and thereby extend the useful life of the corresponding second layer (600a, 600b) (e.g., enabling use of the corresponding second layer (600a, 600b) through a greater number of blending cycles than the second layer (600) when used with process chamber (450)).

[0127] As shown in FIG. 8, first alternative process chamber (450a) is substantially asymmetrical (e.g., vertically asymmetrical) relative to second layer (600a), with upper process chamber portion (550a) being sized and shaped substantially differently from lower process chamber portion (750a). For example, upper process chamber portion (550a) may have a length, a maximum width, a minimum width, and a height defined between lower surface (504a) and ceiling (555a) and substantially less than the maximum width of upper process chamber portion (550a); and lower process chamber portion (750a) may have a length substantially equal to the length of upper process chamber portion (550a), a maximum width substantially equal to the maximum width of upper process chamber portion (550a), and a height defined between upper surface (702a) and floor (755a) and substantially greater than the height of upper process chamber portion (550a). As a result, upper process chamber portion (550a) may be configured to hold a first volume of fluid and lower process chamber portion (750a) may be configured to hold a second volume of fluid substantially greater than the first volume of fluid for upper process chamber portion (550a), such that lower process chamber portion (750a) may have an individual capacity’ substantially greater than that of upper process chamber portion (550a). For example, lower process chamber portion (750a) may have an individual capacity substantially equal to about 75% of the full capacity of first alternative reaction chamber (450a), while upper process chamber portion (550a) may have an individual capacity substantially equal to about 25% of the full capacity of first alternative reaction chamber (450a). In addition, or alternatively, the individual capacity of lower process chamber portion (750a) may be substantially equal to or greater than a predetermined percentage of the full capacity of first alternative reaction chamber (450a) to which the "wet" region of first alternative reaction chamber (450a) is to be filled with an initial amount of fluid at the initiation of a blending cycle. For example, in scenarios where the "wet" region of first alternativereaction chamber (450a) is to be filled with an initial amount of fluid that is substantially equal to about 75% of the full capacity of first alternative reaction chamber (450a). the individual capacity of lower process chamber portion (750a) may be substantially equal to or greater than about 75% of the full capacity of first alternative reaction chamber (450a).

[0128] In the example shown, sidewall (553a) is generally curved (e.g., concave) and ceiling (555a) is generally flat (e.g., horizontal), such that upper process chamber portion (550a) has agenerally flat-topped dome-shaped cross-section with smooth (e.g.. rounded) transitions between sidewall (553a) and ceiling (555a). Such smooth transitions may promote conformation of second layer (600a) with si de wall (553a) and ceiling (555a) when second layer (600a) is deflected into upper process chamber portion (550a) against sidewall (553a) and ceiling (555a), and thereby provide second layer (600a) with a reduced tendency to yield. In the example shown, sidewall (753a) is generally flat (e.g., tapered) and floor (755a) is generally curved (e.g., concave), such that low er process chamber portion (750a) has a generally round-bottomed cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (753a) and floor (755a). Such smooth transitions may promote conformation of second layer (600a) with sidewall (753a) and floor (755a) when second layer (600a) is deflected into lower process chamber portion (750a) against sidewall (753a) and floor (755a), and thereby provide second layer (600a) with a reduced tendency to yield.

[0129] As noted above, the height of upper process chamber portion (550a) is substantially less than the maximum width of upper process chamber portion (550a). The decreased height of upper process chamber portion (550a) relative to the maximum width of upper process chamber portion (550a) may provide the portion of second layer (600a) that extends below upper process chamber portion (550a) and that is configured to be deflected upwardly into upper process chamber portion (550a) with a substantially small stretch ratio within upper process chamber portion (550a). In this regard, the stretch ratio within upper process chamber portion (550a) may be defined as the ratio of the surface area of the portion of second layer (600a) deflected fully u w ardly into upper process chamber portion (550a) vs. the surface area of the same portion undeflected (e.g.. horizontal); and may be expressed as a function of the height of upperprocess chamber portion (550a). the inverse of the maximum width of upper process chamber portion (550a), and the inverse of the length of upper process chamber portion (550a). For example, the stretch ratio may be estimated via the following equation: R = 1 + 2(H / L) + 2(H / W); where R is the stretch ratio, H is the height of upper process chamber portion (550a), W is the maximum width of upper process chamber portion (550a), and L is the length of upper process chamber portion (550a). Thus, the stretch ratio may scale proportionally to the height of upper process chamber portion (550a) and may scale inversely proportionally to the maximum width and / or length of upper process chamber portion (550a).

[0130] For example, within upper process chamber portion (550a), second layer (600a) may have a stretch ratio that is substantially less than about 800%, such as substantially less than about 700%, such as substantially less than about 600%, such as substantially less than about 500%, such as substantially less than about 400%, such as substantially less than about 300%. In some cases, within upper process chamber portion (550a), second layer (600a) may have a stretch ratio that ranges from about 120% to about 450%. For example, within upper process chamber portion (550a), second layer (600a) may have a stretch ratio of about 207% (whereas, within lower process chamber portion (750a), second layer (600a) may have a substantially larger stretch ratio such as about 421%, for example). Such a substantially small stretch ratio within upper process chamber portion (550a) may minimize stretching of second layer (600a) within upper process chamber portion (550a), and thereby provide second layer (600a) with a reduced tendency to yield. Providing second layer (600a) with an increased thickness relative to second layer (600) may also provide second layer (600a) with a reduced tendency to yield. For example, providing second layer (600a) with a thickness of about 200 pm may provide second layer (600a) with a reduced tendency to yield, at least by comparison to second layer (600) described above. It will be appreciated that providing second layer (600a) with an increased thickness may cause a corresponding increase in the amount of pneumatic pressure needed to deflect second layer (600a).

[0131] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (550a) may be substantially equal to the upper process chamberportion length (LUP) of reaction chamber (450); the minimum and maximum widths of upper process chamber portion (550a) may be substantially greater than the upper process chamber portion minimum and maximum widths (Wup,min, Wup.max) of reaction chamber (450), respectively; the height of upper process chamber portion (550a) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750a) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (750a) may be substantially greater than the lower process chamber portion maximum width (Wro,max) of reaction chamber (450); and the height of lower process chamber portion (750a) may be substantially equal to the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550a) may have an individual capacity substantially less than the individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750a) may have an individual capacity substantially greater than the individual capacity of lower process chamber portion (750).

[0132] Thus, upper process chamber portion (550a) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600a) when second layer (600a) is deflected into upper process chamber portion (550a) against sidewall (553a) and ceiling (555a), and thereby provide second layer (600a) with a reduced tendency to yield. In this regard, second layer (600a) may have a stretch ratio within upper process chamber portion (550a) (e.g., about 207%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750a) (e.g.. about 421%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0133] The substantial asymmetry of first alternative reaction chamber (450a), including the increased capacity' of lower process chamber portion (750a) relative to upper process chamber portion (550a), may allow for reduced deflecting of second layer (600a) within first alternative reaction chamber (450a) and thereby provide second layer(600a) with a reduced tendency to yield. More particularly, the increased capacity of lower process chamber portion (750a) relative to upper process chamber portion (550a) may allow second layer (600a) to be substantially undeflected (e.g., flat and / or horizontal) when the “wet” region of first alternative reaction chamber (450a) is filled with an initial amount of fluid at the initiation of a blending cycle. For example, byhaving an individual capacity that is substantially equal to or greater than the predetermined percentage of the full capacity of first alternative reaction chamber (450a) to which the “wet” region of first alternative reaction chamber (450a) is filled with the initial amount of fluid at the initiation of a blending cycle, lower process chamber portion (750a) may hold the entire initial amount of fluid. Thus, second layer (600a) may not be deflected partially upwardly into upper process chamber portion (550a) toward ceiling (555a), and thus may' not undergo any stretching, to accommodate the initial amount of fluid in the “wet” region of first alternative reaction chamber (450a), both in instances where the fluid is substantially compressible and in instances where the fluid is substantially incompressible (e.g., where the fluid has a high water content). In other words, second layer (600a) may be substantially flat when the “wet” region of first alternative reaction chamber (450a) is nominally filled at the initiation of a blending cycle. This functionality of first alternative process chamber (450a) is described in greater detail below in connection with FIGS. 13A-13C.

[0134] As shown in FIG. 8. second alternative process chamber (450b) is substantially asymmetrical (e.g., vertically asymmetrical) relative to second layer (600a), with upper process chamber portion (550b) being sized and shaped substantially differently from lower process chamber portion (750b). For example, upper process chamber portion (550b) may have a length, a maximum width, a minimum width, and a height defined between lower surface (504a) and ceiling (555a) and substantially less than the maximum width of upper process chamber portion (550b); and lower process chamber portion (750b) may have a length substantially equal to the length of upper process chamber portion (550b), a maximum width substantially equal to the maximum width of upper process chamber portion (550b), and a height defined between upper surface (702a) and floor (755a) and substantially greater than the height of upper process chamber portion (550b). As a result, upper process chamber portion (550b)may be configured to hold a first volume of fluid and lower process chamber portion (750b) may be configured to hold a second volume of fluid substantially greater than the first volume of fluid for upper process chamber portion (550b), such that lower process chamber portion (750b) may have an individual capacity substantially greater than that of upper process chamber portion (550b).

[0135] In the example shown, sidewall (553b) is generally curved (e.g., concave) and ceiling (555b) is generally flat (e.g., horizontal), such that upper process chamber portion (550b) has a generally flat-topped dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (553b) and ceiling (555b). Such smooth transitions may promote conformation of second layer (600a) with sidewall (553b) and ceiling (555b) when second layer (600a) is deflected into upper process chamber portion (550b) against sidewall (553b) and ceiling (555b), and thereby provide second layer (600a) with a reduced tendency to yield. In the example shown, sidewall (753b) is generally flat (e.g., tapered) and floor (755b) is generally curved (e.g., concave), such that low er process chamber portion (750b) has a generally round-bottomed cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (753b) and floor (755b). Such smooth transitions may promote conformation of second layer (600a) with sidewall (753b) and floor (755b) when second layer (600a) is deflected into lower process chamber portion (750b) against sidewall (753b) and floor (755b), and thereby provide second layer (600a) with a reduced tendency to yield.

[0136] As noted above, the height of upper process chamber portion (550b) is substantially less than the maximum width of upper process chamber portion (550b). The decreased height of upper process chamber portion (550b) relative to the maximum width of upper process chamber portion (550b) may provide the portion of second layer (600a) that extends below upper process chamber portion (550b) and that is configured to be deflected upwardly into upper process chamber portion (550b) with a substantially small stretch ratio within upper process chamber portion (550b). In this regard, the stretch ratio within upper process chamber portion (550b) may be defined as the ratio of the surface area of the portion of second layer (600a) deflected fully upwardly into upper process chamber portion (550b) vs. the surface area of the same portion undeflected (e.g.. horizontal); and may be expressed as a function of the height of upperprocess chamber portion (550b), the inverse of the maximum width of upper process chamber portion (550b), and the inverse of the length of upper process chamber portion (550b). For example, the stretch ratio may be estimated via the following equation: R = 1 + 2(H / L) + 2(H / W); where R is the stretch ratio, H is the height of upper process chamber portion (550b), W is the maximum width of upper process chamber portion (550b), and L is the length of upper process chamber portion (550b). Thus, the stretch ratio may scale proportionally to the height of upper process chamber portion (550b) and may scale inversely proportionally to the maximum width and / or length of upper process chamber portion (550b).

[0137] For example, within upper process chamber portion (550b), second layer (600a) may have a stretch ratio that is substantially less than about 800%, such as substantially less than about 700%, such as substantially less than about 600%, such as substantially less than about 500%, such as substantially less than about 400%, such as substantially less than about 300%. In some cases, within upper process chamber portion (550b), second layer (600a) may have a stretch ratio that ranges from about 120% to about 450%. For example, within upper process chamber portion (550b), second layer (600a) may have a stretch ratio of about 173% (whereas, within lower process chamber portion (750b), second layer (600a) may have a substantially larger stretch ratio such as about 319%, for example). Such a substantially small stretch ratio within upper process chamber portion (550b) may minimize stretching of second layer (600a) within upper process chamber portion (550b), and thereby provide second layer (600a) with a reduced tendency to yield.

[0138] By way of comparison to reaction chamber (450). the length of upper process chamber portion (550b) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the minimum and maximum w idths of upper process chamber portion (550b) may be substantially greater than the upper process chamber portion minimum and maximum widths (Wup,min, Wup,max) of reaction chamber (450), respectively; the height of upper process chamber portion (550b) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750b) may be substantially equal to the lower process chamber portion length (LLO) of reactionchamber (450); the maximum width of lower process chamber portion (750b) may be substantially greater than the lower process chamber portion maximum width (WLo.max) of reaction chamber (450); and the height of lower process chamber portion (750b) may be substantially less than the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550b) may have an individual capacity’ substantially less than the individual capacity’ of upper process chamber portion (550); and / or lower process chamber portion (750b) may have an individual capacity substantially greater than the individual capacity of lower process chamber portion (750).

[0139] Thus, upper process chamber portion (550b) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600a) when second layer (600a) is deflected into upper process chamber portion (550b) against sidewall (553b) and ceiling (555b), and thereby provide second layer (600a) with a reduced tendency to yield; and / or lower process chamber portion (750b) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600a) when second layer (600a) is deflected into lower process chamber portion (750b) against sidewall (753b) and floor (755b), and thereby provide second layer (600a) with a reduced tendency to yield. In this regard, second layer (600a) may have a stretch ratio within upper process chamber portion (550b) (e.g.. about 173%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750b) (e.g., about 319%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0140] The substantial asymmetry of second alternative reaction chamber (450b), including the increased capacity of lower process chamber portion (750b) relative to upper process chamber portion (550b), may allow for reduced deflecting of second layer (600a) within second alternative reaction chamber (450b) and thereby provide second layer (600a) with a reduced tendency to yield. More particularly, the increased capacity of lower process chamber portion (750b) relative to upper process chamber portion (550b) may allow second layer (600a) to be substantially undeflected (e.g., flatand / or horizontal) when the "w et" region of second alternative reaction chamber (450b) is filled with an initial amount of fluid at the initiation of a blending cycle. For example, by having an individual capacity that is substantially equal to or greater than the predetermined percentage of the full capacity of second alternative reaction chamber (450b) to which the “wet” region of second alternative reaction chamber (450b) is filled with the initial amount of fluid at the initiation of a blending cycle, lower process chamber portion (750b) may hold the entire initial amount of fluid. Thus, second layer (600a) may not be deflected partially upwardly into upper process chamber portion (550b) toward ceiling (555b), and thus may not undergo any stretching, to accommodate the initial amount of fluid in the “wet” region of second alternative reaction chamber (450b). both in instances where the fluid is substantially compressible and in instances where the fluid is substantially incompressible (e.g., where the fluid has a high water content). In other words, second layer (600a) may be substantially flat when the “wet” region of second alternative reaction chamber (450b) is nominally filled at the initiation of a blending cycle.

[0141] As shown in FIG. 8, third alternative process chamber (450c) is generally symmetrical (e.g.. vertically symmetrical) relative to second layer (600a) (e.g.. notwithstanding the presence of pneumatic channels (522a) in first layer (500a) and sidewall channels (754a) in third layer (700a)), with upper process chamber portion (550c) being sized and shaped substantially similarly to lower process chamber portion (750c). For example, upper process chamber portion (550c) may have a length, a maximum width, and a height defined between lower surface (504a) and ceiling (555a); and lower process chamber portion (750c) may have a length substantially equal to the length of upper process chamber portion (550c), a maximum width substantially equal to the maximum width of upper process chamber portion (550c), and a height defined between upper surface (702a) and floor (755a) and substantially equal to the height of upper process chamber portion (550c). As a result, upper process chamber portion (550c) may be configured to hold a first volume of fluid and lower process chamber portion (750c) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (550c). such that each process chamber portion (550c, 750c) of third alternative reaction chamber (450c) mayhave an individual capacity substantially equal to about 50% of the full capacity of third alternative reaction chamber (450c).

[0142] In the example shown, sidewall (553c) is generally flat (e.g., tapered) and ceiling (555c) is generally curved (e.g., concave), such that upper process chamber portion (550c) has a generally round-topped cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (553c) and ceiling (555c). Such smooth transitions may promote conformation of second layer (600a) with sidewall (553 c) and ceiling (555c) when second layer (600a) is deflected into upper process chamber portion (550c) against sidewall (553c) and ceiling (555c), and thereby provide second layer (600a) with a reduced tendency to yield. In the example shown, sidewall (753c) is generally flat (e.g., tapered) and floor (755c) is generally curved (e.g., concave), such that lower process chamber portion (750c) has a generally round-bottomed cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (753c) and floor (755c). Such smooth transitions may promote conformation of second layer (600a) with sidewall (753c) and floor (755c) when second layer (600a) is deflected into lower process chamber portion (750c) against sidewall (753c) and floor (755c), and thereby provide second layer (600a) with a reduced tendency to yield.

[0143] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (550c) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the maximum width of upper process chamber portion (550c) may be substantially greater than the upper process chamber portion maximum width (Wup,max) of reaction chamber (450); the height of upper process chamber portion (550c) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750c) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (750c) may be substantially greater than the lower process chamber portion maximum width (W o.max) of reaction chamber (450); and the height of lower process chamber portion (750c) may be substantially less than the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550c) may have an individual capacity substantially equal tothe individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750c) may have an individual capacity substantially equal to the individual capacity of lower process chamber portion (750).

[0144] Thus, upper process chamber portion (550c) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600a) when second layer (600a) is deflected into upper process chamber portion (550c) against sidewall (553c) and ceiling (555c), and thereby provide second layer (600a) with a reduced tendency to yield; and / or lower process chamber portion (750c) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600a) when second layer (600a) is deflected into lower process chamber portion (750c) against sidewall (753c) and floor (755c), and thereby provide second layer (600a) with a reduced tendency to yield. In this regard, second layer (600a) may have a stretch ratio within upper process chamber portion (550c) (e.g., about 304%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750c) (e.g.. about 304%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0145] As show n in FIG. 8, fourth alternative process chamber (450d) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (600a) (e.g., notwithstanding the presence of pneumatic channels (522a) in first layer (500a) and sidewall channels (754a) in third layer (700a)), with upper process chamber portion (550d) being sized and shaped substantially similarly to lower process chamber portion (750d). For example, upper process chamber portion (550d) may have a length, a maximum width, and a height defined between low er surface (504a) and ceiling (555a); and low er process chamber portion (750d) may have a length substantially equal to the length of upper process chamber portion (550d), a maximum width substantially equal to the maximum width of upper process chamber portion (550d), and a height defined between upper surface (702a) and floor (755a) and substantially equal to the height of upper process chamber portion (550d). As a result, upper process chamber portion (550d) may be configured to hold a first volume of fluid and lower process chamberportion (750d) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (550d). such that each process chamber portion (550d, 750d) of fourth alternative reaction chamber (450d) may have an individual capacity substantially equal to about 50% of the full capacity of fourth alternative reaction chamber (450d).

[0146] In the example shown, sidewall (553d) is generally flat (e.g., tapered) and ceiling (555d) is generally curved (e.g., concave), such that upper process chamber portion (550d) has a generally round-topped cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (553d) and ceiling (555d). Such smooth transitions may promote conformation of second layer (600a) with sidewall (553d) and ceiling (555d) when second layer (600a) is deflected into upper process chamber portion (550d) against sidewall (553d) and ceiling (555d), and thereby provide second layer (600a) with a reduced tendency to yield. In the example shown, sidewall (753d) is generally flat (e.g., tapered) and floor (755d) is generally curved (e.g., concave), such that low er process chamber portion (750d) has a generally round-bottomed cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (753d) and floor (755d). Such smooth transitions may promote conformation of second layer (600a) with sidewall (753d) and floor (755d) when second layer (600a) is deflected into lower process chamber portion (750d) against sidewall (753d) and floor (755d), and thereby provide second layer (600a) with a reduced tendency to yield.

[0147] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (550d) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the maximum width of upper process chamber portion (550d) may be substantially equal to the upper process chamber portion maximum width (Wup,max) of reaction chamber (450); the height of upper process chamber portion (550d) may be substantially equal to the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750d) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (750d) may be substantially equal to the lower process chamber portion maximum width (WLo,max) of reaction chamber (450); and the height of lowerprocess chamber portion (750d) may be substantially equal to the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550d) may have an individual capacity substantially equal to the individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750d) may have an individual capacity substantially equal to the individual capacity of lower process chamber portion (750).

[0148] Due to the more rounded configuration of fourth alternative reaction chamber (450d). second layer (600a) may have a stretch ratio within upper process chamber portion (550d) (e.g., about 498%) that is at least slightly less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750d) (e.g., about 498%) that is at least slightly less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0149] As shown in FIG. 9, fifth alternative process chamber (450e) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (600b) (e.g., notwithstanding the presence of pneumatic channels (522b) in first layer (500b) and sidewall channels (754b) in third layer (700b)), with upper process chamber portion (550e) being sized and shaped substantially similarly to lower process chamber portion (750e). For example, upper process chamber portion (550e) may have a fist length, a maximum width, a minimum width, and a height defined between lower surface (504b) and ceiling (555b); and lower process chamber portion (750e) may have a length substantially equal to the length of upper process chamber portion (550e), a maximum width substantially equal to the maximum width of upper process chamber portion (550e), a minimum width substantially equal to the minimum width of upper process chamber portion (550e), and a height defined between upper surface (702b) and floor (755b) and substantially equal to the height of upper process chamber portion (550e). As a result, upper process chamber portion (550e) may be configured to hold a first volume of fluid and lower process chamber portion (750e) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (550e), such that each process chamber portion (550e, 750e) of fifth alternative reaction chamber (45 Oe) may have an individual capacitysubstantially equal to about 50% of the full capacity of fifth alternative reaction chamber (450e).

[0150] In the example shown, sidewall (553e) is generally curved (e.g., concave) and ceiling (555e) is generally flat (e.g.. horizontal), such that upper process chamber portion (550e) has agenerally flat-topped dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (553e) and ceiling (555e). Such smooth transitions may promote conformation of second layer (600b) with sidewall (553e) and ceiling (555e) when second layer (600b) is deflected into upper process chamber portion (550e) against sidewall (553e) and ceiling (555e), and thereby provide second layer (600b) with a reduced tendency to yield. In the example shown, sidewall (753e) is generally curved (e.g.. concave) and floor (755e) is generally flat (e.g., horizontal), such that lower process chamber portion (750e) has a generally flat-bottomed domeshaped cross-section with smooth (e.g., rounded) transitions between sidewall (753e) and floor (755e). Such smooth transitions may promote conformation of second layer (600b) with sidew all (753e) and floor (755e) when second layer (600b) is deflected into lower process chamber portion (750e) against sidewall (753e) and floor (755e), and thereby provide second layer (600b) with a reduced tendency to yield.

[0151] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (550e) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the maximum width of upper process chamber portion (550e) may be substantially greater than the upper process chamber portion maximum width (Wup,max) of reaction chamber (450); the height of upper process chamber portion (550e) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750e) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (750e) may be substantially greater than the lower process chamber portion maximum width (W o.max) of reaction chamber (450); and the height of lower process chamber portion (750e) may be substantially less than the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550e) may have an individual capacity substantially equal tothe individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750e) may have an individual capacity substantially equal to the individual capacity of lower process chamber portion (750).

[0152] Thus, upper process chamber portion (550e) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into upper process chamber portion (550e) against sidewall (553e) and ceiling (555e), and thereby provide second layer (600b) with a reduced tendency to yield; and / or lower process chamber portion (750e) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into lower process chamber portion (750e) against sidewall (753e) and floor (755e), and thereby provide second layer (600b) with a reduced tendency to yield. In this regard, second layer (600b) may have a stretch ratio within upper process chamber portion (550e) (e.g., about 304%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750e) (e.g.. about 304%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0153] As shown in FIG. 9, sixth alternative process chamber (4501) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (600b) (e.g., notwithstanding the presence of pneumatic channels (522b) in first layer (500b) and sidewall channels (754b) in third layer (700b)), with upper process chamber portion (55 Of) being sized and shaped substantially similarly to lower process chamber portion (750f). For example, upper process chamber portion (550f) may have a length, a maximum width, a minimum width, and a height defined betw een low er surface (504b) and ceiling (555b); and lower process chamber portion (750f) may have a length substantially equal to the length of upper process chamber portion (550f), a maximum width substantially equal to the maximum width of upper process chamber portion (550f), a minimum width substantially equal to the minimum width of upper process chamber portion (550f), and a height defined between upper surface (702b) and floor (755b) and substantially equal to the height of upper process chamber portion (5501).As a result, upper process chamber portion (550f) may be configured to hold a first volume of fluid and lower process chamber portion (7501) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (5501), such that each process chamber portion (550f, 750f) of sixth alternative reaction chamber (450f) may have an individual capacity substantially equal to about 50% of the full capacity of sixth alternative reaction chamber (4501).

[0154] In the example shown, sidewall (5531) is generally curved (e.g., concave) and ceiling (555f) is generally flat (e.g.. horizontal), such that upper process chamber portion (5501) has a generally flat-topped dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (5531) and ceiling (5551). Such smooth transitions may promote conformation of second layer (600b) with sidewall (5531) and ceiling (5551) when second layer (600b) is deflected into upper process chamber portion (5501) against sidewall (5531) and ceiling (5551), and thereby provide second layer (600b) with a reduced tendency to yield. In the example shown, sidewall (7531) is generally curved (e.g., concave) and floor (7551) is generally flat (e.g., horizontal), such that lower process chamber portion (750f) has a generally flat-bottomed dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (7531) and floor (7551). Such smooth transitions may promote conformation of second layer (600b) with sidewall (753f) and floor (7551) when second layer (600b) is deflected into lower process chamber portion (7501) against sidewall (7531) and floor (7551), and thereby provide second layer (600b) with a reduced tendency to yield.

[0155] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (55 Of) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the maximum width of upper process chamber portion (5501) may be substantially greater than the upper process chamber portion maximum width (Wup,max) of reaction chamber (450); the height of upper process chamber portion (5501) may be substantially less than the upper process chamber portion height (Hi p) of reaction chamber (450); the length of lower process chamber portion (750f) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (7501) may be substantially greater than the lower process chamber portion maximumwidth (WLO,max) of reaction chamber (450); and the height of lower process chamber portion (75 Of) may be substantially less than the lower process chamber portion height (Hi.o) of reaction chamber (450). In some versions, upper process chamber portion (550f) may have an individual capacity substantially equal to the individual capacity of upper process chamber portion (550); and / or lower process chamber portion (7501) may- have an individual capacity substantially equal to the individual capacity of lower process chamber portion (750).

[0156] Thus, upper process chamber portion (5501) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into upper process chamber portion (550f) against sidewall (5531) and ceiling (5551), and thereby provide second layer (600b) with a reduced tendency to yield; and / or lower process chamber portion (75 Of) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into lower process chamber portion (750f) against sidewall (7531) and floor (755f), and thereby provide second layer (600b) with a reduced tendency to yield. In this regard, second layer (600b) may have a stretch ratio within upper process chamber portion (550f) (e.g., about 280%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%): and / or may have a stretch ratio within lower process chamber portion (750f) (e.g., about 280%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0157] As shown in FIG. 9. seventh alternative process chamber (450g) is generally symmetrical (e g., vertically symmetrical) relative to second layer (600b) (e.g., notwithstanding the presence of pneumatic channels (522b) in first layer (500b) and sidewall channels (754b) in third layer (700b)), with upper process chamber portion (550g) being sized and shaped substantially similarly to lower process chamber portion (750g). For example, upper process chamber portion (550g) may have a length, a maximum width, a minimum width, and a height defined betw een low er surface (504b) and ceiling (555b); and lower process chamber portion (750g) may have a length substantially equal to the length of upper process chamber portion (550g), a maximumwidth substantially equal to the maximum width of upper process chamber portion (550g). a minimum width substantially equal to the minimum width of upper process chamber portion (550g), and a height defined between upper surface (702b) and floor (755b) and substantially equal to the height of upper process chamber portion (550g). As a result, upper process chamber portion (550g) may be configured to hold a first volume of fluid and lower process chamber portion (750g) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (550g), such that each process chamber portion (550g, 750g) of seventh alternative reaction chamber (450g) may have an individual capacity7substantially equal to about 50% of the full capacity of seventh alternative reaction chamber (450g).

[0158] In the example shown, sidewall (553g) is generally curved (e.g., concave) and ceiling (555g) is generally flat (e.g., horizontal), such that upper process chamber portion (550g) has a generally flat-topped dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (553g) and ceiling (555g). Such smooth transitions may promote conformation of second layer (600b) with sidewall (553g) and ceiling (555g) when second layer (600b) is deflected into upper process chamber portion (550g) against sidewall (553g) and ceiling (555g), and thereby provide second layer (600b) with a reduced tendency to yield. In the example shown, sidewall (753g) is generally curved (e.g.. concave) and floor (755g) is generally flat (e.g., horizontal), such that lower process chamber portion (750g) has a generally flat-bottomed domeshaped cross-section with smooth (e.g., rounded) transitions between sidewall (753g) and floor (755g). Such smooth transitions may promote conformation of second layer (600b) with sidew all (753g) and floor (755g) when second layer (600b) is deflected into lower process chamber portion (750g) against sidewall (753g) and floor (755g), and thereby provide second layer (600b) with a reduced tendency to yield.

[0159] By w ay of comparison to reaction chamber (450), the length of upper process chamber portion (550g) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the maximum width of upper process chamber portion (550g) may be substantially greater than the upper process chamber portion maximum width (Wup,max) of reaction chamber (450); the height of upperprocess chamber portion (550g) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750g) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the maximum width of lower process chamber portion (750g) may be substantially greater than the lower process chamber portion maximum width (WLo,max) of reaction chamber (450); and the height of lower process chamber portion (750g) may be substantially less than the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550g) may have an individual capacity substantially equal to the individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750g) may have an individual capacity substantially equal to the individual capacity of lower process chamber portion (750).

[0160] Thus, upper process chamber portion (550g) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into upper process chamber portion (550g) against sidewall (553g) and ceiling (555g), and thereby provide second layer (600b) with a reduced tendency to yield; and / or lower process chamber portion (750g) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into lower process chamber portion (750g) against sidewall (753g) and floor (755g), and thereby provide second layer (600b) with a reduced tendency to yield. In this regard, second layer (600b) may have a stretch ratio within upper process chamber portion (550g) (e.g., about 257%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750g) (e.g.. about 257%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0161] As shown in FIG. 9, eighth alternative process chamber (450h) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (600b) (e.g., notwithstanding the presence of pneumatic channels (522b) in first layer (500b) and sidewall channels (754b) in third layer (700b)), with upper process chamber portion(550h) being sized and shaped substantially similarly to lower process chamber portion (750h). For example, upper process chamber portion (550h) may have a length, a maximum width, a minimum width, and a height defined between lower surface (504b) and ceiling (555b); and lower process chamber portion (750h) may have a length substantially equal to the length of upper process chamber portion (550h), a maximum width substantially equal to the maximum width of upper process chamber portion (550h), a minimum width substantially equal to the minimum width of upper process chamber portion (550h), and a height defined between upper surface (702b) and floor (755b) and substantially equal to the height of upper process chamber portion (550h). As a result, upper process chamber portion (550h) may be configured to hold a first volume of fluid and lower process chamber portion (750h) may be configured to hold a second volume of fluid substantially equal to the first volume of fluid for upper process chamber portion (550h), such that each process chamber portion (550h, 750h) of eighth alternative reaction chamber (450h) may have an individual capacity substantially equal to about 50% of the full capacity of eighth alternative reaction chamber (450h).

[0162] By way of comparison to reaction chamber (450). the length of upper process chamber portion (550h) may be substantially equal to the upper process chamber portion length (Lup) of reaction chamber (450); the minimum and maximum w idths of upper process chamber portion (550h) may be substantially greater than the upper process chamber portion minimum and maximum widths (Wup,min, Wup,max) of reaction chamber (450), respectively; the height of upper process chamber portion (55 Oh) may be substantially less than the upper process chamber portion height (HUP) of reaction chamber (450); the length of lower process chamber portion (750h) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450): the minimum and maximum widths of lower process chamber portion (750h) may be substantially greater than the lower process chamber portion minimum and maximum widths (WLo,min, WLo,max) of reaction chamber (450), respectively; and the height of lower process chamber portion (750h) may be substantially less than the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, upper process chamber portion (550h) may have an individual capacitysubstantially equal to the individual capacity of upper process chamber portion (550); and / or lower process chamber portion (750h) may have an individual capacity substantially equal to the individual capacity' of lower process chamber portion (750).

[0163] Thus, upper process chamber portion (550h) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into upper process chamber portion (550h) against sidewall (553h) and ceiling (555h), and thereby provide second layer (600b) with a reduced tendency to yield; and / or lower process chamber portion (750h) may be substantially more shallow than lower process chamber portion (750), which may provide reduced stretching of second layer (600b) when second layer (600b) is deflected into lower process chamber portion (750h) against sidewall (753h) and floor (755h), and thereby provide second layer (600b) with a reduced tendency to yield. In this regard, second layer (600b) may have a stretch ratio within upper process chamber portion (550h) (e.g., about 234%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (750h) (e.g.. about 234%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0164] As noted above, any one or more of process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h) may be incorporated into process chip (400) in place of any one or more of process chambers (450, 460, 470). For example, a pair of any of process chambers (450a, 450b, 450c, 450d, 450e, 450f, 450g, 450h), such as a pair of first alternative reaction chambers (450a). may be incorporated into process chip (400) in place of the pair of reaction chambers (450).

[0165] B. Example of Process Chip with Asymmetrical Chambers

[0166] FIGS. 10-14 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. In this regard, in combination with the rest of system (100), process chip (1400) may be utilized to provide in-vitro synthesis, purification, concentration, formulation(e.g., encapsulation of a therapeutic composition in a delivery vehicle through a mixing process, etc.), and / or analysis of therapeutic compositions and / or templates, including but not limited to therapeutic polynucleotides templates, therapeutic polynucleotides, and / or therapeutic polynucleotide compositions. Alternatively, process chip (1400) may be utilized in other processes.

[0167] As shown in FIG. 10, 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).

[0168] As shown in FIG. 10, process chip (1400) of this example further includes a plurality of process chambers (1450, 1460, 1470) that may be used to serve different purposes during the process of producing the therapeutic composition as described herein. By way of example only, process chambers (1450, 1460, 1470) 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). In the present example, process chambers (1450) are configured to be used for IVT reaction, and thus may also be referred to as "‘reaction chambers;” process chambers (1460) are configured to be used for DNA digestion, and thus may also be referred to as “digestion chambers;” and process chambers (1470) are configured to be used for RNA purification, and thus may also be referred to as “purification chambers.” Process chip (1400) may have any other suitable number and / or type(s) of process chambers (1450, 1460. 1470). As shown, process chip (1400) of this example further includes a first layer (1500), a second layer (1600), a third layer (1700), and a fourth layer (1800). Each of these layers (1500, 1600, 1700, 1800), and their structural and functional relationships with each other, will be described in greater detail below'.

[0169] First layer (1500) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, first layer (1500) may be substantially translucent to visible light and / or ultraviolet light. By “substantially translucent” is meant that at least 50% (e.g., at least 90%, including in some instances 100%) of light is transmittedthrough the material compared to a translucent material. First layer (1500) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, first layer (1500) is formed as a lamination of two or more layers of material, such that first layer (1500) does not necessarily need to be formed as a single homogenous continuum of material.

[0170] As shown in FIG. 12, first layer (1500) includes an upper surface (1502) and a lower surface (1504). First layer (1500) may include openings formed through both surfaces (1502, 1504). and such openings may form part of fluid ports (1410) and / or pressure ports (1420). A plurality of process chamber portions (1550, 1560, 1570) are formed as recesses in lower surface (1504), providing respective protruding regions (1552) in upper surface (1502). Each process chamber portion (1550, 1560, 1570) forms a “dry” region of a respective process chamber (1450, 1460, 1470) as described herein.

[0171] Each process chamber portion (1550, 1560, 1570) has a sidewall (1553, 1563, 1573) that extends upwardly from lower surface (1504) toward a corresponding ceiling (1555, 1565, 1575). As seen in FIG. 10, lower surface (1504) of first layer (1500) further defines a plurality of pneumatic channels (1522), which are formed as recesses in lower surface (1504). Each pneumatic channel (1522) is in pneumatic communication with a corresponding opening forming a respective pressure port (1420). Each pneumatic channel (1522) is further configured to define a space between first layer (1500) and second layer (1600), such that pressurized gas may be communicated along pneumatic channels (1522). Each process chamber portion (1550, 1560. 1570) has its own associated pneumatic channel (1522) and pressure port (1420) in this example. Thus, each process chamber portion (1550, 1560, 1570) may be pneumatically pressurized independently of the other process chamber portions (1550, 1560. 1570).

[0172] Lower surface (1504) of first layer (1500) further defines a plurality' of valve chamber portions (1530) and a plurality of pump chamber portions (1540). Valve chamber portions (1530) and pump chamber portions (1540) are formed as recesses in lower surface (1504); and are in pneumatic communication with correspondingpneumatic channels (1522). Pressurized gas may thus be communicated from pressure ports (1420) to valve chamber portions (1530) and pump chamber portions (1540) via respective pneumatic channels (1522).

[0173] Valve chamber portions (1530) form “dry” regions of valve chambers as described herein; while pump chamber portions (1540) form “dry” regions of pump chambers as described herein. In the present example, valve chamber portions (1530) have a circular shape while some pump chamber portions (1540) have an oblong or stadium shape and other pump chamber portions (1540) have a circular shape, though other shapes may be used. By way of further example only, valve chambers associated with valve chamber portions (1530) may be operated like chambers (320, 324) described above; while pump chambers associated with pump chamber portions (1540) may be operated like chamber (322) described above.

[0174] Second layer (1600) is interposed between first layer (1500) and third layer (1700). Second layer (1600) may be in the form of first and second liquid-impermeable flexible membranes laminated to each other in a vertically stacked arrangement. As described below, second layer (1600) may be used to drive fluids across process chip (1400) via peristaltic pumping action. As also described below, second layer (1600) may be used to provide valves at various locations along process chip (1400). By way of example only, one or both flexible membranes of second layer (1600) may comprise polydimethylsilicone (PDMS) elastomer film. Alternatively, any other suitable material(s) may be used to form second layer (1600).

[0175] Second layer (1600) may be configured and operable like elastic layer (1302) described above. Second layer (1600) may thus pass through chambers collectively defined by first and third layers (1500, 1700) to separate such chambers into an upper chamber region (which receives pneumatic pressure) and a lower chamber region (which receives fluid). Such chambers include process chambers (1450, 1460, 1470), valve chambers as described herein, and pump chambers as described herein. Second layer (1600) may further thus be pneumatically deformed to provide valving, peristaltic pumping, etc. within process chip (1400). Second layer (1600) may include a plurality of openings formed through the flexible membranes. The openings may be positionedat fluid ports (1410) and / or pressure ports (1420) to allow fluid and / or pressurized gas to be communicated through second layer (1600) at fluid ports (1410) and / or pressure ports (1420), respectively. The distal ends of fluid and pressure lines (206, 208) may seal against corresponding regions of second layer (1600) at or near the bottom ends of fluid and pressure ports (1410, 1420), respectively.

[0176] In some instances, second layer (1600) may have a substantially increased thickness, at least relative to second layer (600) described above. For example, second layer (1600) may have a thickness of about 200 pm. By way of further example only, second layer (1600) may be configured and operable in accordance with at least some of the teachings of International Pat. App. No. PCT / US2024 / 020488, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,” filed on March 19, 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 entirety7. For example, as best shown in FIG. 14, second layer (1600) may include first and second liquid-impermeable flexible membranes (1600a, 1600b) laminated to each other in a vertically stacked arrangement, and the second flexible membrane (1600b) may include a plurality of apertures (e g., cutouts) (1640) that are each configured to vertically align with a respective chamber (e.g., the pump chamber shown in FIG. 14 defined by pump chamber portions (1540, 1740)), while the first flexible membrane (1600a) may lack such apertures. In such cases, the first flexible membrane (1600a) may have a substantially increased thickness, at least relative to the first flexible membrane of second layer (600) described above. For example, the first flexible membrane (1600a) may have a thickness of about 200 pm. It will be appreciated that any other suitable thickness may be used for second layer (1600) and / or for the first flexible membrane (1600a) of second layer (1600), such as any suitable thickness greater than about 200 pm.

[0177] In the present example, the region of each chamber in process chip (1400) that is above second layer (1600) constitutes a "dry" chamber region since this region receives pressurized gas to pneumatically deflect second layer (1600). Conversely, the region of each chamber in process chip (1400) that is below second layer (1600)constitutes a “wet"’ chamber region since this region receives fluid. However, these "wet" and “dry” roles may be reversed in some variations. In other words, some variations of process chip (1400) may provide pressurized gas to the 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.

[0178] Third layer (1700) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, third layer (1700) is substantially translucent to visible tight and / or ultraviolet tight. Third layer (1700) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, third layer (1700) is formed as a lamination of two or more layers of material, such that third layer (1700) does not necessarily need to be formed as a single homogenous continuum of material.

[0179] As shown in FIG. 12, third layer (1700) includes an upper surface (1702) and a lower surface (1704). Third layer (1700) may include openings formed through both surfaces (1702, 1704), and such openings may form part of fluid ports (1410) and / or pressure ports (1420). A plurality of process chamber portions (1750, 1760, 1770) are formed as recesses in upper surface (1702), providing respective protruding regions (1752) in lower surface (1704). Each process chamber portion (1750. 1760, 1770) forms a “wet” region of a respective process chamber (1450, 1460, 1470) as described herein.

[0180] Each process chamber portion (1750, 1760, 1770) has a sidewall (1753, 1763, 1773) that extends downwardly from upper surface (1702) toward a corresponding floor (1755. 1765, 1775). As seen in FIG. 10, upper surface (1702) of third layer (1700) further defines a plurality of fluid channels (1712), which are formed as recesses in upper surface (1702). Each fluid channel (1712) is in fluid communication with a corresponding opening forming a respective fluid port (1410). Each fluid channel (1712) is further configured to define a space between third layer (1700) and second layer (1600), such that fluid may be communicated along fluid channels (1712). Each process chamber portion (1750, 1760, 1770) also has at least one respective fluid channel (1712) leading thereto, such that fluid may becommunicated to / from each process chamber (1450, 1460, 1470) via the fluid channel (1712) leading to the process chamber portion (1750. 1760, 1770) associated with the process chamber (1450, 1460, 1470). Corresponding pumps and valves interposed between an opening forming a respective fluid port (1410) and each fluid channel (1712) leading to each respective process chamber portion (1750, 1760, 1770) may drive and regulate the communication of fluid to / from each process chamber (1450, 1460, 1470).

[0181] Upper surface (1702) of third layer (1700) further defines a plurality of valve chamber portions (1730) and a plurality of pump chamber portions (1740). Valve chamber portions (1730) and pump chamber portions (1740) are formed as recesses in upper surface (1702); and are in fluid communication with corresponding fluid channels (1712). Fluid may thus be communicated from fluid ports (1410) to valve chamber portions (1730) and pump chamber portions (1740) via respective openings (1710) and fluid channels (1712).

[0182] Valve chamber portions (1730) form “wet” regions of valve chambers as described herein (opposite to a corresponding valve chamber portion (1530)); while pump chamber portions (1740) form ‘‘wet” regions of pump chambers (opposite to a corresponding pump chamber portion (1540)) as described herein. In the present example, valve chamber portions (1730) have a circular shape while some pump chamber portions (1740) have an oblong or stadium shape and other pump chamber portions (1740) have a circular shape, though other shapes may be used. By way of further example only, valve chambers associated with valve chamber portions (1730) may be operated like chambers (320, 324) described above; while pump chambers associated with pump chamber portions (1740) may be operated like chamber (322) described above.

[0183] As also seen in FIGS. 10-12, third layer (1700) includes a plurality of bridging channels (1751), sidewall channels (1754), and floor channels (1756). Sidewall channels (1754) may also be referred to as “sippers.” Each bridging channel (1751) is formed as a recess in upper surface (1702); each floor channel (1756) is formed as a recess in a corresponding floor (1755); and each sidewall channel (1754)is formed as a recess in a corresponding sidewall (1753) that extends upwardly from the corresponding floor channel (1756) toward upper surface (1702). The presence of bridging channels (1751), sidewall channels (1754), and / or floor channels (1756) may provide a pathway for fluid to escape the respective process chamber (1450, 1460, 1470) when second layer (1600) is deformed downwardly toward the respective floor (1755), such that sidewall channels (1754) and / or floor channels (1756) may prevent pockets of fluid from being trapped between localized regions of deformed second layer (1600) and the respective floor (1755) and / or sidewall (1753) (or otherwise reduce the risk of such pockets being formed).

[0184] As shown, at least some sidewall channels (1754) extend from the corresponding floor channel (1756) to a corresponding bridging channel (1751). such that each bridging channel (1751) is flanked by a pair of adjacent sidewall channels (1754). Each bridging channel (1751) and the pair of adjacent sidewall channels (1754) are configured to provide a path for communication of fluid from one process chamber (1450, 1460, 1470) to an adjacent process chamber (1450, 1460, 1470).

[0185] Referring to FIG. 12, fourth layer (1800) may include an elastomeric membrane, such as PDMS or other silicone compositions and / or any other suitable material(s). Fourth layer (1800) may include a central opening that is sized and configured to receive protruding region (1752) of third layer (1700). Fourth layer (1800) may provide a gasket forming a seal between adjacent regions of third layer (1700) and another layer such as a rigid plate. It will be appreciated that any suitable structures or techniques may be used to secure layers (1500, 1600, 1700, 1800) together.

[0186] In the example shown, each upper process chamber portion (1570) is sized and shaped substantially similarly to the corresponding lower process chamber portion (1770), such that each process chamber (1470) is generally symmetrical (e.g., vertically symmetrical) relative to second layer (1600) (e.g., notwithstanding the presence of sidewall channels (1754), floor channels (1756), and bridging channels (1751) in third layer (1700)); while each upper process chamber portion (1550, 1560) is sized and shaped substantially differently from the corresponding lower process chamber portion (1750, 1760), such that each process chamber (1450, 1460) is substantiallyasymmetrical (e.g., vertically asymmetrical) relative to second layer (1600). For example, with specific regard to reaction chambers (1450), each upper process chamber portion (1550) has an upper process chamber portion length (LUP), an upper process chamber portion maximum width (Wup,max), an upper process chamber portion minimum width (Wup,min), and an upper process chamber portion height (HUP) defined between lower surface (1504) and the respective ceiling (1555) and substantially less than the upper process chamber portion maximum width (Wup,max); and each lower process chamber portion (1750) has a lower process chamber portion length (LLO) substantially equal to the upper process chamber portion length (LUP), a lower process chamber portion maximum width (WL0,max) substantially equal to the upper process chamber portion maximum width (Wup.max), and a lower process chamber portion height (HLO) defined between upper surface (1702) and the respective floor (1755) and substantially greater than the upper process chamber portion height (Hup). Thus, reaction chambers (1450) may each be configured and operable similarly to first alternative reaction chamber (450a).

[0187] As a result, each upper process chamber portion (1550) may be configured to hold a first volume of fluid and lower process chamber portion (1750) may be configured to hold a second volume of fluid substantially greater than the first volume of fluid for the corresponding upper process chamber portion (1550), such that each lower process chamber portion (1750) may have an individual capacity substantially greater than that of the corresponding upper process chamber portion (1550). For example, each lower process chamber portion (1750) may have an individual capacity substantially equal to about 75% of the full capacity of the respective reaction chamber (1450), while each upper process chamber portion (1550) may have an individual capacity substantially equal to about 25% of the full capacity of the respective reaction chamber (1450). In addition, or alternatively, the individual capacity of each lower process chamber portion (1750) may be substantially equal to or greater than a predetermined percentage of the full capacity of the respective reaction chamber (1450) to which the “wet’" region of the respective reaction chamber (1450) is to be filled with an initial amount of fluid at the initiation of a blending cycle. For example, in scenarios where thec'wet” region of each reaction chamber (1450) is to be filled with an initialamount of fluid that is substantially equal to about 75% of the full capacity of each reaction chamber (1450). the individual capacity of each lower process chamber portion (1750) may be substantially equal to or greater than about 75% of the full capacity of the respective reaction chamber (1450).

[0188] In the example shown, each sidewall (1553) is generally curved (e.g., concave) and each ceiling (1555) is generally flat (e.g., horizontal), such that each upper process chamber portion (1550) has a generally flat-topped dome-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (1553) and ceiling (1555). Such smooth transitions may promote conformation of second layer (1600) with each sidewall (1553) and each ceiling (1555) when second layer (1600) is deflected into the corresponding upper process chamber portion (1550) against sidewall (1553) and ceiling (1555), and thereby provide second layer (1600) with a reduced tendency to yield. In the example shown, each sidewall (1753) is generally flat (e.g., tapered) and each floor (1755) is generally curved (e.g., concave), such that each lower process chamber portion (1750) has a generally round-bottomed cone-shaped cross-section with smooth (e.g., rounded) transitions between sidewall (1753) and floor (1755). Such smooth transitions may promote conformation of second layer (1600) with each sidewall (1753) and each floor (1755) when second layer (1600) is deflected into the corresponding lower process chamber portion (1750) against sidewall (1753) and floor (1755), and thereby provide second layer (1600) with a reduced tendency to yield.

[0189] As noted above, the upper process chamber portion height (Hup) is substantially less than the upper process chamber portion maximum width (Wup,max). The decreased upper process chamber portion height (HUP) relative to the upper process chamber portion maximum width (Wup.max) may provide the portion of second layer (1600) that extends below upper process chamber portion (1550) and that is configured to be deflected upwardly into upper process chamber portion (1550) with a substantially small stretch ratio within upper process chamber portion (1550). In this regard, the stretch ratio within upper process chamber portion (1550) may be defined as the ratio of the surface area of the portion of second layer (1600) deflected fully upwardly into upper process chamber portion (1550) vs. the surface area of the same portion undeflected (e.g., horizontal); and may be expressed as a function of the upper processchamber portion height (Hup), the inverse of the upper process chamber portion maximum width (Wup.max), and the inverse of the upper process chamber portion length (LI ). For example, the stretch ratio may be estimated via the following equation: R = 1 + 2(HUP / LUP) + 2(HUP / Wup,max). Thus, the stretch ratio may scale proportionally to the upper process chamber portion height (Hup) and may scale inversely proportionally to the upper process chamber portion maximum width (Wup,max) and / or upper process chamber portion length (LUP).

[0190] For example, within upper process chamber portion (1550), second layer (1600) may have a stretch ratio that is substantially less than about 800%, such as substantially less than about 700%, such as substantially less than about 600%, such as substantially less than about 500%, such as substantially less than about 400%, such as substantially less than about 300%. In some cases, within upper process chamber portion (1550), second layer (1600) may have a stretch ratio that ranges from about 120% to about 450%. For example, within upper process chamber portion (1550), second layer (1600) may have a stretch ratio of about 192% (whereas, within lower process chamber portion (1750), second layer (1600) may have a substantially larger stretch ratio such as about 391%, for example). Such a substantially small stretch ratio within upper process chamber portion (1550) may minimize stretching of second layer (1600) within upper process chamber portion (1550), and thereby provide second layer (1600) with a reduced tendency to yield.

[0191] By way of comparison to reaction chamber (450), the upper process chamber portion length (LUP) of each reaction chamber (1450) may be substantially equal to the upper process chamber portion length (LIT) of reaction chamber (450); the upper process chamber portion minimum and maximum widths (Wup,min, Wup,max) of each reaction chamber (1450) may be substantially greater than the upper process chamber portion minimum and maximum widths (Wup,min, Wup,max) of reaction chamber (450), respectively; the upper process chamber portion height (HUP) of each reaction chamber (1450) may be substantially less than the upper process chamber portion height (HI ) of reaction chamber (450); the lower process chamber portion length (LLO) of each reaction chamber (1450) may be substantially equal to the lower process chamber portion length (LLO) of reaction chamber (450); the lower process chamber portionmaximum width (WLO,max) of each reaction chamber (1450) may be substantially greater than the lower process chamber portion maximum width (WL0,max) of each reaction chamber (450); and the lower process chamber portion height (Hi.o) of each reaction chamber (1450) may be substantially equal to the lower process chamber portion height (HLO) of reaction chamber (450). In some versions, each upper process chamber portion (1550) may have an individual capacity substantially less than the individual capacity of upper process chamber portion (550); and / or each lower process chamber portion (1750) may have an individual capacity substantially greater than the individual capacity' of lower process chamber portion (750).

[0192] Thus, each upper process chamber portion (1550) may be substantially more shallow than upper process chamber portion (550), which may provide reduced stretching of second layer (1600) when second layer (1600) is deflected into the corresponding upper process chamber portion (1550) against sidewall (1553) and ceiling (1555), and thereby provide second layer (1600) with a reduced tendency to yield. In this regard, second layer (1600) may have a stretch ratio within upper process chamber portion (1550) (e.g., about 192%) that is substantially less than the stretch ratio of second layer (600) within upper process chamber portion (550) (e.g., about 500%); and / or may have a stretch ratio within lower process chamber portion (1750) (e.g., about 391%) that is substantially less than the stretch ratio of second layer (600) within lower process chamber portion (750) (e.g., about 500%).

[0193] The substantial asymmetry' of each reaction chamber (1450), including the increased capacity of each lower process chamber portion (1750) relative to the corresponding upper process chamber portion (1550), may allow for reduced deflecting of second layer (1600) within the respective reaction chamber (1450) and thereby provide second layer (1600) with a reduced tendency to yield. More particularly, the increased capacity of each lower process chamber portion (1750) relative to the corresponding upper process chamber portion (1550) may allow second layer (1600) to be substantially undeflected (e.g., flat and / or horizontal) when the “wet” region of each reaction chamber (1450) is fdled with an initial amount of fluid at the initiation of a blending cycle. For example, by having an individual capacity that is substantially equal to or greater than the predetermined percentage of the full capacity of therespective reaction chamber (1450) to which the “wet"’ region of the respective reaction chamber (1450) is filled with the initial amount of fluid at the initiation of a blending cycle, each lower process chamber portion (1750) may hold the entire initial amount of fluid. Thus, second layer (1600) may not be deflected partially upwardly into the corresponding upper process chamber portion (1550) toward ceiling (1555), and thus may not undergo any stretching, to accommodate the initial amount of fluid in the “wet” region of the respective reaction chamber (1450), both in instances where the fluid is substantially compressible and in instances where the fluid is substantially incompressible (e.g., where the fluid has a high water content). In other words, second layer (1600) may be substantially flat when the “wet” region of the respective reaction chamber (1450) is nominally filled at the initiation of a blending cycle.

[0194] Referring now primarily to FIGS. 13A-13C, an example of a blending cycle may be performed using process chip (1400). As shown in FIG. 13A, the blending cycle initiates when the “w et” region of each reaction chamber (1450) is filled with an initial amount of fluid that is substantially equal to or less than the individual capacity7of the corresponding lower process chamber portion (1750). For example, the “wet” region of each reaction chamber (1450) may be filled with an initial amount of fluid that is substantially equal to about 75% of the full capacity7of the respective reaction chamber (1450), which may be substantially equal to the individual capacity7of each lower process chamber portion (1750). Thus, second layer (600) is substantially undeflected (e.g., flat and / or horizontal) at the initiation of the blending cycle.

[0195] With the “wet” region of each reaction chamber (1450) filled with the initial amount of fluid, second layer (1600) may be pneumatically deflected downwardly in a first one of the reaction chambers (1450) (e.g., the lefthand reaction chamber (1450) in FIGS. 13A-13C) toward the respective floor (1755) and upwardly in a second one of the reaction chambers (1450) (e.g., the righthand reaction chamber (1450) in FIGS. 13A-13C) toward the respective ceiling (1555). to thereby drive a portion of the fluid from the first reaction chamber (1450) into the second reaction chamber (1450), as show n in FIG. 13B. For example, the portion of the fluid may be driven from the first reaction chamber (1450), through the corresponding bridging channels (1751), and into the second reaction chamber (1450). In some versions, the portion of the fluid drivenfrom the first reaction chamber (1450) into the second reaction chamber (1450) may cause the total amount of fluid within the first reaction chamber (1450) to be substantially equal to about 50% of the full capacity of the first reaction chamber (1450); and / or may cause the total amount of fluid within the second reaction chamber (1450) to be substantially equal to about 100% of the full capacity of the second reaction chamber (1450). Thus, when second layer (1600) is pneumatically deflected downwardly in the first reaction chamber (1450) and upwardly in the second reaction chamber (1450) to drive the portion of the fluid from the first reaction chamber (1450) into the second reaction chamber (1450), second layer (1600) may be deflected only partially downwardly into the corresponding lower process chamber portion (1750) for the first reaction chamber (1450) such that second layer (1600) does not contact the respective floor (1755), and may be deflected fully upwardly into the corresponding upper process chamber portion (1550) for the second reaction chamber (1450) such that second layer (1600) contacts the respective ceiling (1555). In such cases, the smooth transitions between sidewall (1553) and ceiling (1555) of the corresponding upper process chamber portion (1550) for the second reaction chamber (1450) may promote conformation of second layer (1600) with the respective sidewall (1553) and ceiling (1555).

[0196] Second layer (1600) may then be pneumatically deflected downw ardly in the second reaction chamber (1450) toward the respective floor (1755) and upwardly in the first reaction chamber (1450) toward the respective ceiling (1555), to thereby drive a portion of the fluid from the second reaction chamber (1450) into the first reaction chamber (1450), as shown in FIG. 13C. For example, the portion of the fluid may be driven from the second reaction chamber (1450), through the corresponding bridging channels (1751), and into the first reaction chamber (1450). In some versions, the portion of the fluid driven from the second reaction chamber (1450) into the first reaction chamber (1450) may cause the total amount of fluid within the first reaction chamber (1450) to be substantially equal to about 100% of the full capacity of the first reaction chamber (1450); and / or may cause the total amount of fluid within the second reaction chamber (1450) to be substantially equal to about 50% of the full capacity of the second reaction chamber (1450). Thus, when second layer (1600) is pneumaticallydeflected downwardly in the second reaction chamber (1450) and upwardly in the first reaction chamber (1450) to drive the portion of the fluid from the second reaction chamber (1450) into the first reaction chamber (1450), second layer (1600) may be deflected only partially downwardly into the corresponding lower process chamber portion (1750) for the second reaction chamber (1450) such that second layer (1600) does not contact the respective floor (1755). and may be deflected fully upwardly into the corresponding upper process chamber portion (1550) for the first reaction chamber (1450) such that second layer (1600) contacts the respective ceiling (1555). In such cases, the smooth transitions between sidewall (1553) and ceiling (1555) of the corresponding upper process chamber portion (1550) for the first reaction chamber (1450) may promote conformation of second layer (1600) with the respective sidewall (1553) and ceiling (1555).

[0197] Second layer (1600) may alternate between being pneumatically deflected downwardly in the first reaction chamber (1450) and upwardly in the second reaction chamber (1450), and being pneumatically deflected downwardly in the second reaction chamber (1450) and upwardly in the first reaction chamber (1450), to thereby drive fluid back and forth between the first and second reaction chambers (1450) to achieve the desired blending.

[0198] As shown in FIG. 14, each upper pump chamber portion (1540) is sized and shaped substantially differently from the corresponding lower pump chamber portion (1740), such that each pump chamber is substantially asymmetrical (e.g., vertically asymmetrical) relative to second layer (1600). For example, with specific regard to the pump chamber shown in FIG. 14. upper pump chamber portion (1540) has an upper pump chamber portion height (Hup), and lower pump chamber portion (1740) has a lower pump chamber portion height (HLO) substantially less than the upper pump chamber portion height (HUP). In the example shown, upper pump chamber portion (1540) has a generally dome-shaped cross-section, while lower pump chamber portion (1740) has a generally flat-bottomed dome-shaped cross-section.

[0199] The substantial asymmetry of the illustrated pump chamber, including the decreased height of lower pump chamber portion (1740) relative to upper processchamber portion (550a), may allow third layer (1700) to have a decreased thickness relative to first layer (1500). For example, third layer (1700) may have a thickness between upper surface (1702) and lower surface (1704) that is less than a thickness of first layer (1500) between upper surface (1502) and lower surface (1504). In addition, or alternatively, the substantial asymmetry of the illustrated pump chamber, including the decreased height of lower pump chamber portion (1740) relative to upper process chamber portion (550a), may allow the bottom of third layer (1700) to be substantially flat, at least along the regions of third layer (1700) that are outward relative to process chambers (1450, 1460, 1470). For example, lower pump chamber portions (1740) may not provide any protruding regions in lower surface (1704), such that lower surface (1704) may extend continuously outwardly from the upper ends of protruding regions (1752) provided by lower process chamber portions (1750, 1760, 1770) to the outer periphery' of third layer (1700). In the example shown, at least some upper pump chamber portions (1540) provide a corresponding protruding (e.g., bulbous) region, such as a bump (1542) in upper surface (1502), which may allow the thickness of first layer (1500) between upper surface (1502) and lower surface (1504) to be substantially equal to the thickness of third layer (1700) betw een upper surface (1702) and low er surface (1704). In addition, or alternatively, the configuration of the illustrated pump chamber may enable process chip (1400) to be substantially flat-bottomed with a substantially continuous thickness, which may promote moldability of process chip (1400); and / or may improve the ability of second layer (1600 to export all of the fluid w ithin the pump chamber.

[0200] VI. Examples of Combinations

[0201] The following examples relate to various non-exhaustive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, noneof the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors. If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.

[0202] Example 1

[0203] A fluidic apparatus comprising: (a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a first height; (b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber having a second height different from the first height; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being configured to be substantially flat when the first lower chamber portion receives the at least one liquid.

[0204] Example 2

[0205] The fluidic apparatus of Example 1 , the second height being greater than the first height.

[0206] Example 3

[0207] The fluidic apparatus of Example 1 , the second height being less than the first height.

[0208] Example 4

[0209] The fluidic apparatus of any of Examples 1 through 3, the first height being less than a maximum width of the first upper chamber portion.

[0210] Example 5

[0211] The fluidic apparatus of any of Examples 1 through 4, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

[0212] Example 6

[0213] The fluidic apparatus of any of Examples 1 through 5, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0214] Example 7

[0215] The fluidic apparatus of any of Examples 1 through 6, the first upper chamber portion having a first fluid capacity, the first lower chamber having a second fluid capacity different from the first fluid capacity.

[0216] Example 8

[0217] The fluidic apparatus of Example 7, the second fluid capacity being greater than the first fluid capacity.

[0218] Example 9

[0219] The fluidic apparatus of any of Examples 1 through 8, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0220] Example 10

[0221] The fluidic apparatus of any of Examples 1 through 9, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

[0222] Example 11

[0223] The fluidic apparatus of Example 10, the process chamber including a reaction chamber.

[0224] Example 12

[0225] The fluidic apparatus of any of Examples 1 through 9, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

[0226] Example 13

[0227] The fluidic apparatus of any of Examples 1 through 12. the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

[0228] Example 14

[0229] The fluidic apparatus of Example 13, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

[0230] Example 15

[0231] The fluidic apparatus of any of Examples 13 through 14, the second upper chamber portion having the first height.

[0232] Example 16

[0233] The fluidic apparatus of any of Examples 13 through 15, the second lower chamber portion having the second height.

[0234] Example 17

[0235] The fluidic apparatus of any of Examples 13 through 16, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0236] Example 18

[0237] The fluidic apparatus of Example 17, the elastic layer being configured to deflect partially downwardly into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0238] Example 19

[0239] The fluidic apparatus of any of Examples 13 through 18, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least a portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0240] Example 20

[0241] The fluidic apparatus of Example 19, the elastic layer being configured to deflect partially downwardly into the second lower chamber portion and to deflect fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0242] Example 21

[0243] A fluidic apparatus comprising: (a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a ceiling and at least one sidewall, the first upper chamber portion having a first fluid capacity; (b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber having a second fluid capacity different from the first fluid capacity; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deflectable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being deflectable into the first upper chamber portion against the ceiling and the at least one sidewall for drawing the at least one liquid into the first lower chamber portion.

[0244] Example 22

[0245] The fluidic apparatus of Example 21, the second fluid capacity being greater than the first fluid capacity.

[0246] Example 23

[0247] The fluidic apparatus of any of Examples 21 through 22, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

[0248] Example 24

[0249] The fluidic apparatus of Example 23, the second height being greater than the first height.

[0250] Example 25

[0251] The fluidic apparatus of any of Examples 23 through 24. the first height being less than a maximum width of the first upper chamber portion.

[0252] Example 26

[0253] The fluidic apparatus of any of Examples 23 through 25. the first height being defined between the ceiling and the elastic layer.

[0254] Example 27

[0255] The fluidic apparatus of any of Examples 23 through 26, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0256] Example 28

[0257] The fluidic apparatus of any of Examples 21 through 27, the second fluid capacity being substantially equal to or greater than about 75% of a sum of the first and second fluid capacities.

[0258] Example 29

[0259] The fluidic apparatus of any of Examples 21 through 28, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0260] Example 30

[0261] The fluidic apparatus of any of Examples 21 through 29, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

[0262] Example 31

[0263] The fluidic apparatus of Example 30, the process chamber including a reaction chamber.

[0264] Example 32

[0265] The fluidic apparatus of any of Examples 21 through 29, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

[0266] Example 33

[0267] The fluidic apparatus of any of Examples 21 through 32, the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

[0268] Example 34

[0269] The fluidic apparatus of Example 33, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

[0270] Example 35

[0271] The fluidic apparatus of any of Examples 33 through 34, the second upper chamber portion having the first fluid capacity.

[0272] Example 36

[0273] The fluidic apparatus of any of Examples 33 through 35, the second lower chamber portion having the second fluid capacity.

[0274] Example 37

[0275] The fluidic apparatus of any of Examples 33 through 36, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0276] Example 38

[0277] The fluidic apparatus of Example 37, the elastic layer being configured to deflect partially downwardly into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0278] Example 39

[0279] The fluidic apparatus of any of Examples 33 through 38, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least a portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0280] Example 40

[0281] The fluidic apparatus of Example 39, the elastic layer being configured to deflect partially downwardly into the second lower chamber portion and to deflect fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0282] Example 41

[0283] A fluidic apparatus comprising: (a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a ceiling and at least one sidewall; (b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber portion and the first upper chamber portion collectively defining a first chamber; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deflectable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being deflectable into the first upper chamber portion against the ceiling and the at least one sidewall for drawing the at least one liquid into the first lower chamber portion, the first chamber being substantially asymmetrical relative to the elastic layer.

[0284] Example 42

[0285] The fluidic apparatus of Example 41, the first upper chamber portion having a first fluid capacity, the second upper chamber portion having a second fluid capacity different from the first fluid capacity, and the first chamber having a total fluid capacity comprising a sum of the first and second fluid capacities.

[0286] Example 43

[0287] The fluidic apparatus of Example 42, the second fluid capacity being substantially equal to or greater than about 75% of the total fluid capacity.

[0288] Example 44

[0289] The fluidic apparatus of any of Examples 41 through 43, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

[0290] Example 45

[0291] The fluidic apparatus of Example 44, the second height being greater than the first height.

[0292] Example 46

[0293] The fluidic apparatus of any of Examples 44 through 45. the first height being less than a maximum width of the first upper chamber portion.

[0294] Example 47

[0295] The fluidic apparatus of any of Examples 44 through 46, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

[0296] Example 48

[0297] The fluidic apparatus of any of Examples 44 through 47, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0298] Example 49

[0299] The fluidic apparatus of any of Examples 41 through 48, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0300] Example 50

[0301] The fluidic apparatus of any of Examples 41 through 49, the first chamber including a process chamber.

[0302] Example 51

[0303] The fluidic apparatus of Example 50, the process chamber including a reaction chamber.

[0304] Example 52

[0305] The fluidic apparatus of any of Examples 41 through 49, the first chamber including at least one of a pump chamber or a valve chamber.

[0306] Example 53

[0307] The fluidic apparatus of any of Examples 41 through 52, the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas. the second layer further defining a secondlower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

[0308] Example 54

[0309] The fluidic apparatus of Example 53, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

[0310] Example 55

[0311] The fluidic apparatus of any of Examples 53 through 54, the second upper chamber portion having the first fluid capacity .

[0312] Example 56

[0313] The fluidic apparatus of any of Examples 53 through 55, the second lower chamber portion having the second fluid capacity7.

[0314] Example 57

[0315] The fluidic apparatus of any of Examples 53 through 56, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0316] Example 58

[0317] The fluidic apparatus of Example 57, the elastic layer being configured to deflect partially downwardly7into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

[0318] Example 59

[0319] The fluidic apparatus of any of Examples 53 through 58, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least aportion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0320] Example 60

[0321] The fluidic apparatus of Example 59, the elastic layer being configured to deflect partially downwardly into the second lower chamber portion and to deflect fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

[0322] Example 61

[0323] A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, the first upper chamber portion having a first fluid capacity’, (ii) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber having a second fluid capacity different from the first fluid capacity, and (iii) an elastic layer disposed betw een the first layer and the second layer, the method comprising: (a) filling the first lower chamber portion with an initial amount of liquid substantially equal to or less than the second fluid capacity, such that the elastic layer is in an undeflected state; and (b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first lower chamber portion to thereby drive at least a portion of the liquid out of the first low er chamber portion.

[0324] Example 62

[0325] The method of Example 61, the second fluid capacity being greater than the first fluid capacity7.

[0326] Example 63

[0327] The method of any of Examples 61 through 62, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

[0328] Example 64

[0329] The method of Example 63, the second height being greater than the first height.

[0330] Example 65

[0331] The method of any of Examples 63 through 64, the first height being less than a maximum width of the first upper chamber portion.

[0332] Example 66

[0333] The method of any of Examples 63 through 65, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

[0334] Example 67

[0335] The method of any of Examples 63 through 66, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0336] Example 68

[0337] The method of any of Examples 61 through 67, the second fluid capacity being substantially equal to or greater than about 75% of a sum of the first and second fluid capacities.

[0338] Example 69

[0339] The method of any of Examples 61 through 68, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0340] Example 70

[0341] The method of any of Examples 61 through 69, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

[0342] Example 71

[0343] The method of Example 70, the process chamber including a reaction chamber.

[0344] Example 72

[0345] The method of any of Examples 61 through 69, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

[0346] Example 73

[0347] The method of any of Examples 61 through 72, the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

[0348] Example 74

[0349] The method of Example 73, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

[0350] Example 75

[0351] The method of any of Examples 73 through 74, the second upper chamber portion having the first fluid capacity.

[0352] Example 76

[0353] The method of any of Examples 73 through 75, the second lower chamber portion having the second fluid capacity.

[0354] Example 77

[0355] The method of any of Examples 73 through 76, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0356] Example 78

[0357] The method of Example 77, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly into the first lower chamber portion while deflecting the elastic layer fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0358] Example 79

[0359] The method of any of Examples 73 through 78, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive at least a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

[0360] Example 80

[0361] The method of Example 79, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

[0362] Example 81

[0363] A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, the first upper chamber portion having a first height, (ii) a second layer defining a first low er chamber portion positioned under the first upper chamber portion, the first lower chamber having a second height different from the first height, and (iii) an elastic layer disposed between the first layer and the second layer, the method comprising: (a) filling the first lower chamber portion with an initial amount of liquid such that the elastic layer is in an undeflected state; and (b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first lower chamber portion to thereby drive at least a portion of the liquid out of the first lower chamber portion.

[0364] Example 82

[0365] The method of Example 81, the second height being greater than the first height.

[0366] Example 83

[0367] The method of Example 81 , the second height being less than the first height.

[0368] Example 84

[0369] The method of any of Examples 81 through 83, the first height being less than a maximum width of the first upper chamber portion.

[0370] Example 85

[0371] The method of any of Examples 81 through 84, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

[0372] Example 86

[0373] The method of any of Examples 81 through 85, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0374] Example 87

[0375] The method of any of Examples 81 through 86, the first upper chamber portion having a first fluid capacity, the first lower chamber having a second fluid capacity different from the first fluid capacity.

[0376] Example 88

[0377] The method of Example 87, the second fluid capacity being greater than the first fluid capacity.

[0378] Example 89

[0379] The method of any of Examples 81 through 88, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0380] Example 90

[0381] The method of any of Examples 81 through 89, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

[0382] Example 91

[0383] The method of Example 90, the process chamber including a reaction chamber.

[0384] Example 92

[0385] The method of any of Examples 81 through 89, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

[0386] Example 93

[0387] The method of any of Examples 81 through 92, the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

[0388] Example 94

[0389] The method of Example 93, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

[0390] Example 95

[0391] The method of any of Examples 93 through 94, the second upper chamber portion having the first height.

[0392] Example 96

[0393] The method of any of Examples 93 through 95, the second lower chamber portion having the second height.

[0394] Example 97

[0395] The method of any of Examples 93 through 96, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0396] Example 98

[0397] The method of Example 97, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly intothe first lower chamber portion while deflecting the elastic layer fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0398] Example 99

[0399] The method of any of Examples 93 through 98, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive at least a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

[0400] Example 100

[0401] The method of Example 99, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second low er chamber portion and into the first low er chamber portion.

[0402] Example 101

[0403] A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, (ii) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion and the first upper chamber portion collectively defining a first chamber, and (iii) an elastic layer disposed between the first layer and the second layer, the first chamber being substantially asymmetrical relative to the elastic layer, the method comprising: (a) filling the first low er chamber portion with an initial amount of liquid such that the elastic layer is in an undeflected state; and (b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first low er chamber portion to thereby drive at least a portion of the liquid out of the first lower chamber portion.

[0404] Example 102

[0405] The method of Example 101, the first upper chamber portion having a first fluid capacity, the second upper chamber portion having a second fluid capacity different from the first fluid capacity, and the first chamber having a total fluid capacity comprising a sum of the first and second fluid capacities.

[0406] Example 103

[0407] The method of Example 102, the second fluid capacity being substantially equal to or greater than about 75% of the total fluid capacity.

[0408] Example 104

[0409] The method of any of Examples 101 through 103, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

[0410] Example 105

[0411] The method of Example 104, the second height being greater than the first height.

[0412] Example 106

[0413] The method of any of Examples 104 through 105, the first height being less than a maximum width of the first upper chamber portion.

[0414] Example 107

[0415] The method of any of Examples 104 through 106, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

[0416] Example 108

[0417] The method of any of Examples 104 through 107, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

[0418] Example 109

[0419] The method of any of Examples 101 through 108. the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

[0420] Example 110

[0421] The method of any of Examples 101 through 109, the first chamber including a process chamber.

[0422] Example 111

[0423] The method of Example 110, the process chamber including a reaction chamber.

[0424] Example 112

[0425] The method of any of Examples 101 through 109, the first chamber including at least one of a pump chamber or a valve chamber.

[0426] Example 113

[0427] The method of any of Examples 101 through 112. the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

[0428] Example 114

[0429] The method of Example 113. the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

[0430] Example 1 15

[0431] The method of any of Examples 1 13 through 114. the second upper chamber portion having the first fluid capacity.

[0432] Example 1 16

[0433] The method of any of Examples 113 through 115. the second lower chamber portion having the second fluid capacity.

[0434] Example 1 17

[0435] The method of any of Examples 113 through 116, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0436] Example 118

[0437] The method of Example 117, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly into the first lower chamber portion while deflecting the elastic layer fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

[0438] Example 119

[0439] The method of any of Examples 113 through 118, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive at least a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

[0440] Example 120

[0441] The method of Example 1 19, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

[0442] VII. Miscellaneous

[0443] The foregoing description is provided to enable a person skilled in the art to practice the various configurations described herein. While the subject technology has been particularly described with reference to the various figures and configurations, it should be understood that these are for illustration purposes only and should not be taken as limiting the scope of the subject technology.

[0444] There may be many other ways to implement the subject technology. Various functions and elements described herein may be partitioned differently from those shown without departing from the scope of the subject technology. Various modifications to these implementations may be readily apparent to those skilled in the art, and generic principles defined herein may be applied to other implementations. Thus, many changes and modifications may be made to the subject technology, by one having ordinary skill in the art, without departing from the scope of the subject technology. For instance, different numbers of a given module or unit may be employed, a different type or types of a given module or unit may be employed, a given module or unit may be added, or a given module or unit may be omitted.

[0445] 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.

[0446] In versions implemented using a computer system, 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. In general, use of the term "input device" is intended to include all possible ty pes of devices and ways to input information into computer system.

[0447] In versions implemented using a 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. In general, use of the term "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.

[0448] In versions implemented using a 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. 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 functionality7of certain implementations may be stored by file storage subsystem in the storage subsystem, or in other machines accessible by the processor.

[0449] In versions implemented using a computer system, 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, aserver farm, a widely-distributed set of loosely networked computers, or any other data processing system or user device. Due to the ever-changing nature of computers and networks, 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.

[0450] As an article of manufacture, rather than a method, a non-transitory computer readable medium (CRM) 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. Alternatively, the program instructions may be loaded on a non-transitor ' CRM and, when combined with appropriate hardware, become a component of one or more of the computer- implemented systems that practice the methods disclosed.

[0451] Underlined and / or italicized headings and subheadings are used for convenience only, do not limit the subject technology, and are not referred to in connection with the interpretation of the description of the subject technology7. All structural and functional equivalents to the elements of the various implementations described throughout this disclosure that are known or later come to be known to those of ordinary skill in the art are expressly incorporated herein by reference and intended to be encompassed by the subject technology7. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the above description.

[0452] It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in greater detail below (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are contemplated as being part of the inventive subject matter disclosed herein.

Claims

WHAT IS CLAIMED IS:

1. A fluidic apparatus comprising:(a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a first height;(b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber having a second height different from the first height; and(c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being configured to be substantially flat when the first lower chamber portion receives the at least one liquid.

2. The fluidic apparatus of claim 1, the second height being greater than the first height.

3. The fluidic apparatus of claim 1, the second height being less than the first height.

4. The fluidic apparatus of any of claims 1 through 3, the first height being less than a maximum width of the first upper chamber portion.

5. The fluidic apparatus of any of claims 1 through 4, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

6. The fluidic apparatus of any of claims 1 through 5, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

7. The fluidic apparatus of any of claims 1 through 6, the first upper chamber portion having a first fluid capacity’, the first lower chamber having a second fluid capacity' different from the first fluid capacity.

8. The fluidic apparatus of claim 7, the second fluid capacity7being greater than the first fluid capacity.

9. The fluidic apparatus of any of claims 1 through 8, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

10. The fluidic apparatus of any of claims 1 through 9, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

11. The fluidic apparatus of claim 10, the process chamber including a reaction chamber.

12. The fluidic apparatus of any of claims 1 through 9, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

13. The fluidic apparatus of any of claims 1 through 12, the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

14. The fluidic apparatus of claim 13. the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

15. The fluidic apparatus of any of claims 13 through 14, the second upper chamber portion having the first height.

16. The fluidic apparatus of any of claims 13 through 15, the second lower chamber portion having the second height.

17. The fluidic apparatus of any of claims 13 through 16, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

18. The fluidic apparatus of claim 17, the elastic layer being configured to deflect partially downwardly into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

19. The fluidic apparatus of any of claims 13 through 18, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least a portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

20. The fluidic apparatus of claim 19, the elastic layer being configured to deflect partially downw ardly into the second low er chamber portion and to deflect fullyupwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

21. A fluidic apparatus comprising:(a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a ceiling and at least one sidewall, the first upper chamber portion having a first fluid capacity;(b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber having a second fluid capacity different from the first fluid capacity’; and(c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deflectable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being deflectable into the first upper chamber portion against the ceiling and the at least one sidewall for drawing the at least one liquid into the first lower chamber portion.

22. The fluidic apparatus of claim 21. the second fluid capacity being greater than the first fluid capacity.

23. The fluidic apparatus of any of claims 21 through 22, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

24. The fluidic apparatus of claim 23, the second height being greater than the first height.

25. The fluidic apparatus of any of claims 23 through 24, the first height being less than a maximum width of the first upper chamber portion.

26. The fluidic apparatus of any of claims 23 through 25, the first height being defined between the ceiling and the elastic layer.

27. The fluidic apparatus of any of claims 23 through 26, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

28. The fluidic apparatus of any of claims 21 through 27, the second fluid capacity being substantially equal to or greater than about 75% of a sum of the first and second fluid capacities.

29. The fluidic apparatus of any of claims 21 through 28, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

30. The fluidic apparatus of any of claims 21 through 29, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

31. The fluidic apparatus of claim 30, the process chamber including a reaction chamber.

32. The fluidic apparatus of any of claims 21 through 29, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

33. The fluidic apparatus of any of claims 21 through 32. the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

34. The fluidic apparatus of claim 33, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

35. The fluidic apparatus of any of claims 33 through 34, the second upper chamber portion having the first fluid capacity.

36. The fluidic apparatus of any of claims 33 through 35, the second lower chamber portion having the second fluid capacity.

37. The fluidic apparatus of any of claims 33 through 36, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

38. The fluidic apparatus of claim 37, the elastic layer being configured to deflect partially downwardly into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

39. The fluidic apparatus of any of claims 33 through 38, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least a portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

40. The fluidic apparatus of claim 39, the elastic layer being configured to deflect partially downwardly into the second lower chamber portion and to deflect fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second low er chamber portion and into the first lower chamber portion.41 . A fluidic apparatus comprising:(a) a first layer defining a first upper chamber portion, the first upper chamber portion being configured to receive pressurized gas, the first upper chamber portion having a ceiling and at least one sidewall;(b) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion being configured to receive at least one liquid, the first lower chamber portion and the first upper chamber portion collectively defining a first chamber; and(c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deflectable into the first lower chamber portion to thereby drive the at least one liquid out of the first lower chamber portion, the elastic layer being deflectable into the first upper chamber portion against the ceiling and the at least one sidewall for drawing the at least one liquid into the first lower chamber portion, the first chamber being substantially asymmetrical relative to the elastic layer.

42. The fluidic apparatus of claim 41. the first upper chamber portion having a first fluid capacity, the second upper chamber portion having a second fluid capacity different from the first fluid capacity, and the first chamber having a total fluid capacity comprising a sum of the first and second fluid capacities.

43. The fluidic apparatus of claim 42, the second fluid capacity being substantially equal to or greater than about 75% of the total fluid capacity.

44. The fluidic apparatus of any of claims 41 through 43, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

45. The fluidic apparatus of claim 44, the second height being greater than the first height.

46. The fluidic apparatus of any of claims 44 through 45, the first height being less than a maximum width of the first upper chamber portion.

47. The fluidic apparatus of any of claims 44 through 46, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

48. The fluidic apparatus of any of claims 44 through 47, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

49. The fluidic apparatus of any of claims 41 through 48, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

50. The fluidic apparatus of any of claims 41 through 49, the first chamber including a process chamber.

51. The fluidic apparatus of claim 50, the process chamber including a reaction chamber.

52. The fluidic apparatus of any of claims 41 through 49, the first chamber including at least one of a pump chamber or a valve chamber.

53. The fluidic apparatus of any of claims 41 through 52. the first layer further defining a second upper chamber portion, the second upper chamber portion being configured to receive the pressurized gas, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion, the second lower chamber portion being configured to receive the at least one liquid.

54. The fluidic apparatus of claim 53, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid between the first and second lower chamber portions.

55. The fluidic apparatus of any of claims 53 through 54, the second upper chamber portion having the first fluid capacity.

56. The fluidic apparatus of any of claims 53 through 55, the second lower chamber portion having the second fluid capacity.

57. The fluidic apparatus of any of claims 53 through 56, the elastic layer being deflectable into the first lower chamber portion to thereby drive at least a portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

58. The fluidic apparatus of claim 57, the elastic layer being configured to deflect partially downw ardly into the first lower chamber portion and to deflect fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the first lower chamber portion and into the second lower chamber portion.

59. The fluidic apparatus of any of claims 53 through 58, the elastic layer being deflectable into the second lower chamber portion to thereby drive at least a portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

60. The fluidic apparatus of claim 59, the elastic layer being configured to deflect partially downw ardly into the second low er chamber portion and to deflect fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the at least one liquid out of the second lower chamber portion and into the first lower chamber portion.

61. A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, the first upper chamber portion having a first fluid capacity, (ii) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber having a second fluid capacity different from the first fluid capacity', and (iii) an elastic layer disposed between the first layer and the second layer, the method comprising:(a) filling the first lower chamber portion with an initial amount of liquid substantially equal to or less than the second fluid capacity, such that the elastic layer is in an undeflected state; and(b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first lower chamber portion to thereby drive at least a portion of the liquid out of the first lower chamber portion.

62. The method of claim 61 , the second fluid capacity being greater than the first fluid capacity.

63. The method of any of claims 61 through 62, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

64. The method of claim 63, the second height being greater than the first height.

65. The method of any of claims 63 through 64, the first height being less than a maximum width of the first upper chamber portion.

66. The method of any of claims 63 through 65, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

67. The method of any of claims 63 through 66, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

68. The method of any of claims 61 through 67, the second fluid capacity being substantially equal to or greater than about 75% of a sum of the first and second fluid capacities.

69. The method of any of claims 61 through 68, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

70. The method of any of claims 61 through 69, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

71. The method of claim 70, the process chamber including a reaction chamber.

72. The method of any of claims 61 through 69, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

73. The method of any of claims 61 through 72, the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

74. The method of claim 73. the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

75. The method of any of claims 73 through 74, the second upper chamber portion having the first fluid capacity.

76. The method of any of claims 73 through 75, the second lower chamber portion having the second fluid capacity.

77. The method of any of claims 73 through 76, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

78. The method of claim 77, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly into the first lower chamber portion while deflecting the elastic layer fully upwardlyinto the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

79. The method of any of claims 73 through 78, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive at least a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

80. The method of claim 79, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

81. A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, the first upper chamber portion having a first height, (ii) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber having a second height different from the first height, and (iii) an elastic layer disposed between the first layer and the second layer, the method comprising:(a) filling the first lower chamber portion with an initial amount of liquid such that the elastic layer is in an undeflected state; and(b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first lower chamber portion to thereby drive at least a portion of the liquid out of the first lower chamber portion.

82. The method of claim 81, the second height being greater than the first height.

83. The method of claim 81, the second height being less than the first height.

84. The method of any of claims 81 through 83, the first height being less than a maximum width of the first upper chamber portion.

85. The method of any of claims 81 through 84, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

86. The method of any of claims 81 through 85. the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

87. The method of any of claims 81 through 86, the first upper chamber portion having a first fluid capacity, the first lower chamber having a second fluid capacity different from the first fluid capacity.

88. The method of claim 87, the second fluid capacity being greater than the first fluid capacity.

89. The method of any of claims 81 through 88, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

90. The method of any of claims 81 through 89, the first upper chamber portion and the first lower chamber portion collectively defining a process chamber.

91. The method of claim 90, the process chamber including a reaction chamber.

92. The method of any of claims 81 through 89, the first upper chamber portion and the first lower chamber portion collectively defining at least one of a pump chamber or a valve chamber.

93. The method of any of claims 81 through 92, the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

94. The method of claim 93, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

95. The method of any of claims 93 through 94, the second upper chamber portion having the first height.

96. The method of any of claims 93 through 95, the second lower chamber portion having the second height.

97. The method of any of claims 93 through 96, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

98. The method of claim 97, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly into the first lower chamber portion while deflecting the elastic layer fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second low er chamber portion.

99. The method of any of claims 93 through 98, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive atleast a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

100. The method of claim 99, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

101. A method of using an apparatus, the apparatus comprising (i) a first layer defining a first upper chamber portion, (ii) a second layer defining a first lower chamber portion positioned under the first upper chamber portion, the first lower chamber portion and the first upper chamber portion collectively defining a first chamber, and (iii) an elastic layer disposed between the first layer and the second layer, the first chamber being substantially asymmetrical relative to the elastic layer, the method comprising:(a) filling the first lower chamber portion with an initial amount of liquid such that the elastic layer is in an undeflected state; and(b) with the first lower chamber portion filled with the initial amount of liquid, deflecting the elastic layer into the first lower chamber portion to thereby drive at least a portion of the liquid out of the first lower chamber portion.

102. The method of claim 101, the first upper chamber portion having a first fluid capacity, the second upper chamber portion having a second fluid capacity different from the first fluid capacity , and the first chamber having a total fluid capacity comprising a sum of the first and second fluid capacities.

103. The method of claim 102, the second fluid capacity being substantially equal to or greater than about 75% of the total fluid capacity.

104. The method of any of claims 101 through 103, the first upper chamber portion having a first height, the first lower chamber portion having a second height different from the first height.

105. The method of claim 104, the second height being greater than the first height.

106. The method of any of claims 104 through 105, the first height being less than a maximum width of the first upper chamber portion.

107. The method of any of claims 104 through 106, the first upper chamber portion having a ceiling, the first height being defined between the ceiling and the elastic layer.

108. The method of any of claims 104 through 107, the first lower chamber portion having a floor, the second height being defined between the floor and the elastic layer.

109. The method of any of claims 101 through 108, the first upper chamber portion having a first cross-sectional shape, the first lower chamber portion having a second cross-sectional shape different from the first cross-sectional shape.

110. The method of any of claims 101 through 109, the first chamber including a process chamber.

111. The method of claim 110. the process chamber including a reaction chamber.

112. The method of any of claims 101 through 109, the first chamber including at least one of a pump chamber or a valve chamber.

113. The method of any of claims 101 through 112, the first layer further defining a second upper chamber portion, the second layer further defining a second lower chamber portion positioned under the second upper chamber portion.

114. The method of claim 113, the second layer further defining at least one bridging channel extending between the first and second lower chamber portions.

115. The method of any of claims 113 through 114, the second upper chamber portion having the first fluid capacity.1 16. The method of any of claims 1 13 through 1 15, the second lower chamber portion having the second fluid capacity.

117. The method of any of claims 113 through 116, the act of deflecting the elastic layer into the first lower chamber portion being performed to thereby drive at least a portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

118. The method of claim 117, the act of deflecting the elastic layer into the first lower chamber portion including deflecting the elastic layer partially downwardly into the first lower chamber portion while deflecting the elastic layer fully upwardly into the second upper chamber portion, to thereby drive at least the portion of the liquid out of the first lower chamber portion and into the second lower chamber portion.

119. The method of any of claims 113 through 118, further comprising deflecting the elastic layer into the second lower chamber portion to thereby drive at least a portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

120. The method of claim 119, the act of deflecting the elastic layer into the second lower chamber portion including deflecting the elastic layer partially downwardly into the second lower chamber portion while deflecting the elastic layer fully upwardly into the first upper chamber portion, to thereby drive at least the portion of the liquid out of the second lower chamber portion and into the first lower chamber portion.

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