EP4731539A1 - Reagent reservoirs and related systems and methods - Google Patents

Abstract

Reagent reservoirs and related systems and methods are disclosed. An apparatus includes a flexible container and a coupling. The flexible container includes an end and a tapered bottom. The flexible container includes an interior containing dried reagent. The coupling has a portion coupled to the end of the flexible container. The coupling has a port fluidly coupled to the interior of the flexible container. Rehydrating fluid is to flow through the port and into the interior of the flexible container. An interaction between the rehydrating fluid and the tapered bottom is to cause a vortex within the interior of the flexible container to rehydrate a substantial portion of the dried reagent.

Description

REAGENT RESERVOIRS AND RELATED SYSTEMS AND METHODS

CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application Number 63/509,980, filed June 23, 2023, the content of which is incorporated by reference herein in its entirety and for all purposes.

BACKGROUND

[0002] Various protocols in biological or chemical research involve performing controlled reactions. The designated reactions can then be observed or detected and subsequent analysis can help identify or reveal properties of chemicals involved in the reaction. In some multiplex assays, an unknown analyte having an identifiable label {e.g., fluorescent label) can be exposed to thousands of known probes under controlled conditions. Each known probe can be deposited into a corresponding well of a microplate. Observing any chemical reactions that occur between the known probes and the unknown analyte within the wells can help identify or reveal properties of the analyte. Other examples of such protocols include known deoxyribonucleic acid (DNA) sequencing processes, such as sequencing-by-synthesis (SBS) or cyclic-array sequencing.

[0003] In some fluorescent-detection protocols, an optical system is used to direct excitation light onto fluorophores, e.g., fluorescently-labeled analytes and to also detect the fluorescent emissions signal light that can emit from the analytes having attached fluorophores. In other proposed detection systems, the controlled reactions in a flow cell are detected by a solid-state light sensor array {e.g., a complementary metal oxide semiconductor (CMOS) detector). These systems do not involve a large optical assembly to detect the fluorescent emissions. For CMOS based flow cells that use external illumination, the lid over the flow channel may be transparent. Furthermore, the external illumination source often is aligned with the sensor, which may have particular challenges for removable flow cells and/or multiple flow cells used in a single instrument.

[0004] Some sequencing, such as DNA sequencing, may include moving reagents, buffers, and/or other materials through a flow channel of one or more flow cells, maintaining and/or modifying the temperature(s) of the materials within the flow channel, and illuminating fluorescent nucleotides within the flow channel. To use a shared pool of reagent resources for each flow cell may involve a fluidic solution that passes fluids to multiple flow cells either simultaneously or on demand. Certain reagents may be unstable or otherwise degrade if stored for extended periods of time. SUMMARY

[0005] Shortcomings of the prior art can be overcome and advantages and benefits as described later in this disclosure can be achieved through the provision of reagent cartridges and related systems and methods. Various implementations of the apparatus and methods are described below, and the apparatus and methods, including and excluding the additional implementations enumerated below, in any combination (provided these combinations are not inconsistent), may overcome these shortcomings and achieve the advantages and benefits described herein.

[0006] In accordance with a first implementation, an an apparatus, comprising: a flexible container comprising an end and a tapered bottom, the flexible container including an interior containing dried reagent; and a coupling having a portion coupled to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container, wherein rehydrating fluid is to flow through the port and into the interior of the flexible container and wherein an interaction between the rehydrating fluid and the tapered bottom is to cause a vortex within the interior of the flexible container to rehydrate a substantial portion of the dried reagent.

[0007] In accordance with a second implementation, a method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; enabling a vortex to form within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom, and a rehydrating the dried reagent in the interior using the vortex.

[0008] In accordance with a third implementation, a method, comprising: forming a tapered bottom of a flexible container; depositing dried reagent within an interior of the flexible container; and coupling a coupling to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container.

[0009] In accordance with a fourth implementation, a method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom, and rehydrating the dried reagent in the interior using the vortex.

[0010] In accordance with a fifth implementation, an apparatus, comprising: a flexible container comprising an end and a tapered bottom. The flexible container including an interior containing dried reagent. A coupling having a portion coupled to the end of the flexible container. The coupling comprising a port fluidly coupled to the interior of the flexible container. Rehydrating fluid is to flow through the port and into the interior of the flexible container and an interaction between the rehydrating fluid and the tapered bottom is to rehydrate a substantial portion of the dried reagent.

[0011] In accordance with a sixth implementation, a method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; breaching the dried reagent with the liquid; and rehydrating the dried reagent in the interior.

[0012] In accordance with a seventh implementation, a method, comprising: forming a tapered bottom of a flexible container; depositing dried reagent within an interior of the flexible container; and coupling securing a coupling to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container.

[0013] In further accordance with the foregoing first, second, third, fourth, fifth, sixth, and seventh implementations, an apparatus and/or method may further comprise or include any one or more of the following:

[0014] In an implementation, the flexible container comprises a heat-sealed pouch.

[0015] In another implementation, the heat-sealed pouch comprises a first panel and a second panel.

[0016] In another implementation, the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

[0017] In another implementation, the tapered bottom is V-shaped.

[0018] In another implementation, the tapered bottom comprises a conical bottom.

[0019] In another implementation, the tapered bottom comprises a vertex.

[0020] In another implementation, the vertex has an angle that is less than 180 degrees.

[0021] In another implementation, the vertex has an angle that is less than 180 degrees.

[0022] In another implementation, the vertex has an angle about 45 degrees and about 105 degrees.

[0023] In another implementation, the vertex has an angle between about 60 degrees and about 90 degrees. [0024] In another implementation, the angle is between about 45 degrees and about 105 degrees.

[0025] In another implementation, the angle is between about 60 degrees and about 90 degrees.

[0026] In another implementation, the angle is about 60 degrees.

[0027] In another implementation, the angle is about 90 degrees.

[0028] In another implementation, the flexible container comprises heat seals that comprise the tapered bottom and form the interior that are triangular.

[0029] In another implementation, the flexible container comprises heat seals that comprise the tapered bottom and form the interior that are pentagonal.

[0030] In another implementation, further comprising a sipper.

[0031] In another implementation, the sipper is integrally formed with the flexible container.

[0032] In another implementation, the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

[0033] In another implementation, the tapered bottom comprises a vertex and the sipper includes a distal end disposed proximate the vertex.

[0034] In another implementation, further comprising a heat seal seam that forms a baffle within the interior.

[0035] In another implementation, the flexible container comprises a second end opposite the end and wherein the baffle extends toward the second end.

[0036] In another implementation, further comprising a first heat seal seam that forms a first baffle within the interior and a second heat seal seam that forms a second baffle within the interior.

[0037] In another implementation, the coupling further comprises an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0038] In another implementation, the extension comprises a tube having a proximal end coupled to the portion of the coupling and a distal end disposed within the interior of the flexible container. [0039] In another implementation, the flexible container comprises lateral sides comprising reinforcements.

[0040] In another implementation, reinforcements of the lateral sides comprise inserts.

[0041] In another implementation, further comprising a frame carried by the flexible container.

[0042] In another implementation, the flexible container comprises heat seal seams forming the interior and lateral sides, wherein a pocket is formed between the heat seal seams, the frame positioned within the pocket.

[0043] In another implementation, the frame enables the flexible container to be substantially taught.

[0044] In another implementation, the frame is U-shaped.

[0045] In another implementation, the flexible container further comprising an external border, the apparatus further comprising a frame engaging the flexible container.

[0046] In another implementation, the frame is selectively receivable between the interior and the external border.

[0047] In another implementation, the frame is U-shaped.

[0048] In another implementation, the interior comprises a volume of approximately 13 milliliters.

[0049] In another implementation, the coupling comprises a fitment.

[0050] In another implementation, the portion of the coupling has opposing first side walls that form a canoe shape.

[0051] In another implementation, further comprising a second flexible container having an end and defining a second interior, the flexible container positioned within the second interior.

[0052] In another implementation, the coupling has the portion coupled to the end of the flexible container and a second portion coupled to the end of the second flexible container, the coupling comprising a pressure port fluidly coupled to the second interior of the second flexible container.

[0053] In another implementation, the reagent coupling is to couple with the port to enable reagent to flow out of the flexible container and the pressure coupling is to enable a pressure source to change a pressure within the second flexible container to urge the reagent to flow out of the flexible container. [0054] In another implementation, the reagent comprises lyophilized microspheres.

[0055] In another implementation, the flexible container and the coupling coupled thereto are vacuum sealed.

[0056] In another implementation, flowing the rehydrating fluid through the port comprises flowing the rehydrating fluid through the port of a coupling having a portion coupled to an end of the flexible container, the coupling comprising the port fluidly coupled to the interior of the flexible container.

[0057] In another implementation, the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

[0058] In another implementation, flowing rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container.

[0059] In another implementation, flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper formed by a pair of heat seal seams.

[0060] In another implementation, flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper comprising an extension coupled to the port and extending into the interior of the flexible container.

[0061] In another implementation, the extension comprises a tube having a proximal end coupled to a portion of a coupling and a distal end disposed within the interior of the flexible container.

[0062] In another implementation, further comprising reducing foaming within the interior of the interior of the flexible container.

[0063] In another implementation, reducing the foaming comprises reducing the foaming using baffles.

[0064] In another implementation, the baffles are formed by heat seal seams.

[0065] In another implementation, the heat seal seams comprise a first heat seal seam extending inwardly from a first lateral side of the flexible container and a second heat seal seam extending inwardly from a second lateral side of the flexible container. [0066] In another implementation, flowing the rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container and between the baffles.

[0067] In another implementation, further comprises encouraging the flexible container to be substantially taut.

[0068] In another implementation, encouraging the flexible container to be substantially taught comprises reinforcing lateral sides of the flexible container.

[0069] In another implementation, the reinforcements of the lateral sides comprise inserts.

[0070] In another implementation, the reinforcements of the lateral sides comprise a frame.

[0071] In another implementation, the frame is a U-shaped frame.

[0072] In another implementation, forming the tapered bottom comprises forming a first heat seal seam and a second heat seal seam.

[0073] In another implementation, further comprising forming a sipper integrally formed with the flexible container.

[0074] In another implementation, the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

[0075] In another implementation, further comprising forming a baffle within the interior.

[0076] In another implementation, further comprising forming a first baffle within the interior and a second baffle within the interior.

[0077] In another implementation, further comprising coupling an extension to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0078] In another implementation, further comprising forming an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0079] In another implementation, further comprising vacuum sealing the flexible container.

[0080] In another implementation, breaching the dried reagent comprises the rehydrating fluid interacting with the tapered bottom. [0081] In another implementation, further comprising forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom.

[0082] In another implementation, rehydrating the dried reagent in the interior comprises rehydrating the dried reagent in the interior using the vortex.

[0083] 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 subject matter disclosed herein and/or may be combined to achieve the particular benefits of a particular aspect. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are contemplated as being part of the subject matter disclosed herein.

BRIEF DESCRIPTION OF DRAWINGS

[0084] The present disclosure is described in the following detailed description in conjunction with the drawings, wherein:

[0085] FIG. 1 illustrates a schematic diagram of an implementation of a system in accordance with the teachings of this disclosure.

[0086] FIG. 2 is a detailed cross-sectional view of a portion of an implementation of a reagent reservoir interface that can be used to implement the reagent reservoir interface of the system of FIG. 1 and a portion of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1.

[0087] FIG. 3 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 .

[0088] FIG. 4 is an isometric view of the coupling of the reagent reservoir of FIG. 3.

[0089] FIG. 5 is a detailed isometric cross-sectional view of the reagent reservoir of FIG. 3.

[0090] FIG. 6 is an isometric view of a plurality of reagent reservoirs of FIG. 3 that are coupled together using a plate.

[0091] FIG. 7 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 .

[0092] FIG. 8 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 . [0093] FIG. 9 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 .

[0094] FIG. 10 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 .

[0095] FIG. 11 is a side view of an implementation of a reagent reservoir that can be used to implement the reagent reservoir of FIG. 1 .

[0096] FIG. 12 illustrates a flow chart for a method of using the reagent reservoirs, the first flexible container, the flexible container, or any of the other implementations disclosed herein.

[0097] FIG. 13 illustrates an example of a bag made in accordance with the present disclosure.

[0098] FIG. 14 illustrates the bag of FIG. 13 partially filled with dried reagent.

[0099] FIG. 15 shows the bag of FIG. 13 providing higher recovery and better uniformity over other bag designs.

[0100] FIG. 16 illustrates an alternative bag, similar to the bag of FIG. 13.

[0101] FIG. 17 illustrates a flexible shell having an integrated sipper, an internal volume, and an external border that include heat seals.

[0102] FIG. 18 illustrates a flexible shell including the sipper, the first internal volume, the second internal volume, and the external border including heat seals.

[0103] FIG. 19 illustrates the bag of FIG. 13 including an external border spaced apart from the internal volume.

[0104] FIG. 20 illustrates a cross-sectional view of the bag of FIG. 13 as the internal volume is filled with dried reagent and/or rehydration fluid.

[0105] FIG. 21 illustrates a cross-sectional view of the bag of FIG. 13 with a first fitment and a second fitment.

[0106] FIG. 22 illustrates the improved recovery and uniformity possible with the bags as disclosed herein.

[0107] FIG. 23 illustrates a bag inefficiently rehydrating dried reagent.

[0108] FIG. 24 illustrates an example of rehydration fluid breaching the dried reagent. DETAILED DESCRIPTION

[0109] Although the following text discloses a detailed description of implementations of methods, apparatuses and/or articles of manufacture, it should be understood that the legal scope of the property right is defined by the words of the claims set forth at the end of this patent. Accordingly, the following detailed description is to be construed as examples only and does not describe every possible implementation, as describing every possible implementation would be impractical, if not impossible. Numerous alternative implementations could be implemented, using either current technology or technology developed after the filing date of this patent. It is envisioned that such alternative implementations would still fall within the scope of the claims.

[0110] Reagents may be lyophilized or dried for stability for extended periods of storage, and then rehydrated prior to use.

[0111] At least some of the examples of the flow cells described herein help enable the storage and rehydration of dried reagents. Some reagents are expensive, and may only be required in small volumes. The small volumes of reagents may also benefit from being rehydrated to high concentration to allow for sufficient stability for further storage in liquid form before use. However, rehydrating dried reagents to high concentration may be difficult. For example, it may be beneficial to rehydrate 12 mL of dried or lyophilized reagent with about 6 ml_ of liquid, such as buffer. The dried reagent may be lyophilized microspheres. A bag may be used to store the lyophilized microspheres. Because the liquid volume is lower than dry reagent volume, not all microsphere may be exposed to liquid depending on the configuration of the container or vessel. In an automated system, the bag may be stationary and not agitated from outside; rather a stationary sipper which has the opening at the bottom of the bag may be utilized. Mixing may be accomplished by moving liquid back and forth from the straw, sometimes referred to as jet mixing.

[0112] The present application is directed to a bag for storing and rehydrating dried reagent. As will be described in greater detail below, the bag comprises a bag configured to facilitate breaching, form rehydrating vortices, maintain intra-bag lyophilized reagent movement. In the present example, the bag is a flexible bag, but in various other examples, the bag may be flexible, semi-flexible, semi-rigid, or rigid.

[0113] FIG. 1 illustrates a schematic diagram of an implementation of a system 100 in accordance with the teachings of this disclosure. The system 100 can be used to perform an analysis on one or more samples of interest. The sample may include one or more DNA clusters that are linearized to form a single stranded DNA (sstDNA). In the implementation shown, the system 100 receives a reagent reservoir 102 and a flow cell 104 and includes, in part, a reagent receptacle 105, a pressure source 106, a reagent reservoir interface 108, a drive assembly 110, a controller 1 12, an imaging system 114, and a waste reservoir 1 16. The reagent reservoir 102 may alternatively be referred to as a reagent vessel, a reagent assembly, or a reagent cartridge. The reagent reservoir 102 contains reagent 118 in the implementation shown and the reagent receptacle 105 receives the reagent reservoir 102. The reagent 1 18 may be dry reagent. The reagent 118 may include lyophilized microspheres. The dry reagent may be referred to as lyophilized reagent or dried reagent. The reagent 1 18 may alternatively be liquid reagent. The reagent reservoir interface 108 has a reagent coupling 120 and a pressure coupling 122 fluidly coupled to the pressure source 106. The pressure coupling 122 may alternatively be omitted.

The controller 112 is electrically and/or communicatively coupled to the pressure source 106, the reagent reservoir interface 108, the drive assembly 1 10, and the imaging system 114 and causes the pressure source 106, the reagent reservoir interface 108, the drive assembly 110, and/or the imaging system 114 to perform various functions as disclosed herein.

[0114] The flow cell 104 carries a sample of interest. The pressure source 106 may be used to pressurize the reagent reservoir 102 to flow the reagent 118 from the reagent reservoir 102 that interact with the sample. The pressure source 106 may be provided by the system 100 and/or may be carried by the reagent reservoir 102. Alternatively, the pressure source 106 may be omitted.

[0115] A reversible terminator may be attached to the reagent 118 to allow a single nucleotide to be incorporated onto a growing DNA strand. One or more of the nucleotides has a unique fluorescent label that emits a color when excited in some implementations. The color (or absence thereof) is used to detect the corresponding nucleotide. The imaging system 1 14 excites one or more of the identifiable labels (e.g., a fluorescent label) and thereafter obtains image data for the identifiable labels. The labels may be excited by incident light and/or a laser and the image data may include one or more colors emitted by the respective labels in response to the excitation. The image data (e.g., detection data) may be analyzed by the system 100. The imaging system 114 may be a fluorescence spectrophotometer including an objective lens and/or a solid-state imaging device. The solid-state imaging device may include a charge coupled device (CCD) and/or a complementary metal oxide semiconductor (CMOS).

[0116] Another reaction component (e.g., a reagent) is flowed into the flow cell 104 after the image data is obtained that is thereafter received by the waste reservoir 109 and/or otherwise exhausted by the reagent reservoir 102. The reaction component may perform a flushing operation that chemically cleaves the fluorescent label and the reversible terminator from the sstDNA. The sstDNA is then ready for another cycle.

[0117] The reagent reservoir 102 has a first flexible container 124, a second flexible container 126, and a coupling 128. The first flexible container 124 may be referred to as the flexible container. The first flexible container 124 may be referred to as a reagent reservoir and the second flexible container 126 may be referred to as a pressure chamber. The second flexible container may alternatively be omitted. The first flexible container 124 has an end 130 and a tapered bottom 200, and defines an interior 132 that contains the reagent 118. The second flexible container 126 also has an end 134 and defines an interior 136. An internal volume of the first interior 132 may be about 13 milliliters. The internal volume of the first interior 132 may be any other volume. The internal volume of the first interior 132 may between about 5 milliliters and about 500 milliliters, for example. The internal volume of the first interior 132 may be less than about 5 milliliters and/or larger than about 500 milliliters in some examples. The internal volume of the first interior 132 may between about 1 milliliters and about 1 Liter, for example. The interior 132 may be referred to as a first interior and the interior 136 may be referred to as a second interior. The first flexible container 124 is positioned within the interior 136 of the second flexible container 126. The second flexible container 126 may alternatively be omitted as shown in FIGS. 7 - 10, for example.

[0118] The first flexible container 124 may be a heat-sealed pouch 135 in some examples. The first flexible container 124 may include a first panel 202 and a second panel 204. The panels 202, 204 are coupled to form the first flexible container 124. The panels 202, 204 may be referred to as sheets. Any number of panels may be used to form the first flexible container 124, however.

[0119] The first flexible container 124 has a first heat seal seam 203 and a second heat seal seam 205 that form the tapered bottom 200 in the implementation shown. The tapered bottom 200 is V-shaped and/or has a conical bottom. The tapered bottom 200 may have another shape, however. The tapered bottom 200 has a vertex 206. The vertex 206 has an angle that is shown being less than 180 degrees. The angle may be between about 45 degrees and about 105 degrees in some examples. The angle may be between about 60 degrees and about 90 degrees. The angle may be about 60 degrees in some examples. The angle may be about 90 degrees in some examples. The tapered bottom 200 may have any angle however that encourages a vortex to formed within the interior of the interior 136 of the first flexible container 124 and/or that increases rehydration of the reagent 1 18 within the first flexible container 124, for example.

[0120] The coupling 128 has a first portion 138 coupled and/or secured to the end 130 of the first flexible container 124 and a second portion 140 coupled to the end 134 of the second flexible container 126. The second portion 140 may be omitted when the second flexible container 126 is omitted, for example. The coupling 128 may be coupled to the first flexible container 124 and the second flexible container 126 using a heat sealing process. The coupling 128 also has a reagent port 142 fluidly coupled to the interior 132 of the first flexible container 124 and a pressure port 144 fluidly coupled to the interior 136 of the second flexible container 126. The reagent coupling 120 couples with the reagent port 142 in operation to enable the reagent 118 to flow out of the first flexible container 124 to the reagent coupling 120 of the reagent reservoir interface 108 and the pressure coupling 122 couples with the pressure port 144 to enable the pressure source 106 to change a pressure within the second flexible container 126 to urge the reagent 118 to flow out of the first flexible container 124. Rehydrating fluid flows through the reagent port 142 and into the interior 132 of the first flexible container 124 in operation. An interaction between the rehydrating fluid and the tapered bottom 200 of the first flexible container 124 causes a vortex within the interior 132 of the first flexible container 124 to rehydrate a substantial portion of the dried reagent.

[0121] The pressure source 106 may provide positive pressure to the second flexible container 126 and/or the pressure source 106 may provide negative pressure to the second flexible container 126. The pressure source 106 changes a pressure within the second flexible container 126 and that pressure change compresses the first flexible container 124 and urges the reagent 118 out of the first flexible container 124 regardless of whether the pressure provided is a positive pressure or a negative pressure. The pressure within the second flexible container 126 may apply a relatively even force on the first flexible container 124. All, a majority, or most of the reagent 118 may be dispensed from the first flexible container 124 as a result and, thus, the reagent reservoirs 102 disclosed have low amounts of dead volume.

[0122] A seal 146 is carried by the reagent port 142 and a seal 148 is carried by the pressure port 144. The seal 146 provides a fluid seal and the seal 148 provides a pressure seal. The reagent coupling 120 sealingly engages the seal 146 carried by the reagent port 142 and the pressure coupling 122 sealingly engages the seal 148 carried by the pressure port 144. The seals 146, 148 may be O-rings and/or may be include an elastomer. [0123] The reagent reservoir 102 also includes a cover 150 that covers the reagent port 142 and the pressure port 144. The cover 150 may be an impermeable barrier 152 such as foil. The cover 150 retains the reagent 118 within the first flexible container 124 and/or prevents or inhibits ingress of moisture into the interior 132 of the first flexible container 124 and/or the interior 136 of the second flexible container 126. The cover 150 may be a pierceable or removable cover including thin metal foil, such as aluminum foil, or a thin plastic sheet(s), such as Saran™ wrap. The cover 150 may comprise or consist of other materials and/or other layering arrangements that substantially prevent moisture ingress. The cover 150 may be coupled to the reagent port 142 and the pressure port 144 by heat sealing, laser welding, ultrasonic welding, pressuresensitive adhesive (PSA), or any other suitable method. The cover 150 may alternatively be omitted. A pierceable septum or resealing spring valve may be included in place of the cover 150, for example.

[0124] The system 100 may pierce the cover 150 or the cover 150 may be pierced by an individual prior to use. The reagent coupling 120 may pierce the cover 150 covering the reagent port 142 in implementations when the system 100 pierces the cover 150 and the pressure coupling 122 may pierce the cover 150 covering the pressure port 144. The reagent coupling 120 and/or the pressure coupling 122 may include a piercing member such as a conical protrusion that is used to pierce the cover 150 as and/or prior to the couplings being formed between the reagent coupling 120 and the reagent port 142 and between the pressure coupling 122 and the pressure port 144.

[0125] A sipper 154 is also included in the implementation shown. The sipper 154 may be integrally formed with the first flexible container 124 in some implementation. In such implementations, the first flexible container 124 may include a heat seal 208 that forms a channel 210 that forms the sipper 154 within the interior 132. The heat seal 208 may include a pair of heat seal seams between which the channel 210 is defined. The sipper 154 may be coupled to the reagent port 142 and extend into the interior 132 of the first flexible container 124. The sipper 154 may alternatively be formed as a tube 155. The tube 155 may be referred to as a straw.

[0126] The sipper 154 may be used to encourage more of the reagent 1 18 to flow out of the first flexible container 124 and, thus, for less dead volume to be left within the first flexible container 124 when emptied. The sipper 154 may alternatively be omitted. The sipper 154 has a proximal end 156 coupled to the first portion 138 of the coupling 128 and a distal end 158 disposed within the interior 132 of the first flexible container 124. The distal end 158 of the sipper 154 is shown disposed proximate the vertex 206. [0127] The sipper 154 may be coupled with the coupling 128 by an interference fit. The coupling 128 may include a male portion that is received within a female portion of the sipper 154, for example. The sipper 154 may be coupled to the coupling 128 in other ways, however. Adhesive may be used to couple the tube 155 and the coupling 128, for example.

[0128] An insert 159 may be positioned within the interior 136 of the second flexible container 126 and positioned between the second flexible container 126 and the first flexible container 124. The insert 159 may be a structure and/or a mesh that allows the second flexible container 126 to slide and/or move relative to the first flexible container 124. Relative movement between the second flexible container 126 and the first flexible container 124 is helpful when the pressure source 106 applies negative pressure.

[0129] A regulator 160 can be positioned between the pressure source 106 and the reagent reservoir interface 108 and regulates a pressure provided to the reagent reservoir interface 108 and, thus, the second flexible container 126. The regulator 160 may alternatively not be included.

[0130] The reagent reservoir 102 is in fluid communication with the flow cell 104. A “flow cell” as used herein can include a device having a lid extending over a reaction structure to form a flow channel therebetween that is in communication with a plurality of reaction sites of the reaction structure, and can include a detection device that detects designated reactions that occur at or proximate to the reaction sites. The flow cell 104 is shown being received within a flow cell receptacle 162 of the system 100. The flow cell 104 may alternatively be carried by or otherwise integrated into the reagent reservoir 102.

[0131] While the above disclosure describes urging liquid and/or the reagent 118 into and out of the first flexible container 124 and/or through the flow cell 104 under positive pressure, liquid and/or the reagent 1 18 may alternatively be drawn through the flow cell 104 under negative pressure when, for example, the reagent reservoir(s) 102 is not pressurized. The system 100 may include a pump 164 positioned between the flow cell 104 and the waste reservoir 116 to do so. The waste reservoir 109 may be selectively receivable within a waste reservoir receptacle 165 of the system 100. The pump 164 may be implemented by a syringe pump, a peristaltic pump, a diaphragm pump, etc. While the pump 164 is shown being part of the system 100 and positioned between the flow cell 104 and the waste reservoir 11 16 the pump 164 may be positioned upstream of the flow cell 104, may be part of the reagent reservoir 102, or omitted entirely, in other implementations. [0132] Referring now to the drive assembly 110, in the implementation shown, the drive assembly 1 10 includes a pump drive assembly 166 that interfaces with the pump 164 to pump fluid through the reagent reservoir 102 and/or the flow cell 104.

[0133] The controller 112 includes a user interface 168, a communication interface 170, one or more processors 172, and a memory 174 storing instructions executable by the one or more processors 172 to perform various functions including the disclosed implementations. The user interface 168, the communication interface 170, and the memory 174 are electrically and/or communicatively coupled to the one or more processors 172.

[0134] In an implementation, the user interface 168 receives input from a user and provides information to the user associated with the operation of the system 100 and/or an analysis taking place. The user interface 168 may include a touch screen, a display, a key board, a speaker(s), a mouse, a track ball, and/or a voice recognition system. The touch screen and/or the display may display a graphical user interface (GUI).

[0135] In an implementation, the communication interface 170 enables communication between the system 100 and a remote system(s) (e.g., computers) via a network(s). The network(s) may include an intranet, a local-area network (LAN), a wide-area network (WAN), the intranet, etc. Some of the communications provided to the remote system may be associated with analysis results, imaging data, etc. generated or otherwise obtained by the system 100. Some of the communications provided to the system 100 may be associated with a fluidics analysis operation, patient records, and/or a protocol(s) to be executed by the system 100.

[0136] The one or more processors 172 and/or the system 100 may include one or more of a processor-based system(s) or a microprocessor-based system(s). In some implementations, the one or more processors 172 and/or the system 100 includes a reduced-instruction set computer(s) (RISC), an application specific integrated ci rcuit(s) (ASICs), a field programable gate array(s) (FPGAs), a field programable logic device(s) (FPLD(s)), a logic circuit(s), and/or another logic-based device executing various functions including the ones described herein.

[0137] The memory 174 can include one or more of a hard disk drive, a flash memory, a readonly memory (ROM), erasable programable read-only memory (EPROM), electrically erasable programable read-only memory (EEPROM), a random-access memory (RAM), non-volatile RAM (NVRAM) memory, a compact disk (CD), a digital versatile disk (DVD), a cache, and/or any other storage device or storage disk in which information is stored for any duration (e.g., permanently, temporarily, for extended periods of time, for buffering, for caching). [0138] FIG. 2 is a detailed cross-sectional view of a portion of an implementation of a reagent reservoir interface 180 that can be used to implement the reagent reservoir interface 108 of the system 100 of FIG. 1 and a portion of a reagent reservoir 182 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir interface 180 includes the reagent coupling 120 and the pressure coupling 122 and the reagent reservoir 182 includes the reagent port 142 and the pressure port 144. The seals 146, 148 are positioned within the corresponding ports 142, 144.

[0139] The reagent coupling 120 is shown being received within the reagent port 142 and the pressure coupling 122 is shown being received within the pressure port 144. The reagent port 142 is formed of or comprises an annular wall 205 and the reagent port 142 is also formed of or comprises an annular wall 207. The reagent coupling 120 includes a reagent conduit 213 having a distal end 209 that is shown received within the reagent port 142 and the pressure coupling 122 similarly includes a pressure conduit 21 1 having a distal end 214 received within the pressure port 144.

[0140] FIG. 3 is a side view of an implementation of a reagent reservoir 300 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 300 includes the first flexible container 124, the second flexible container 126, and the coupling 128. The first flexible container 124 includes heat seals 212.

[0141] The first flexible container 124 is a pouch bag 302 in the implementation shown and the second flexible container 126 is also a pouch bag 304. The pouch bag 302 is a 3-sided pouch having opposing side surfaces 306 and a base 307 and the pouch bag 304 is also shown as a 3- sided pouch having opposing side surfaces 308 and a base 309. The pouch bags 302, 304 may be differently configured, however. The pouch bags 302, 304 may be 2-sided bags or 4-sided bags, for example.

[0142] The first flexible container 124 has a neck 310 and the first portion 138 of the coupling 128 is coupled to the neck 310 of the first flexible container 124. The second flexible container 126 also has a neck 312 and the second portion 140 of the coupling 128 is coupled to the neck 312 of the second flexible container 126. The first flexible container 124 and the second flexible container 126 are shown being symmetric about a central longitudinal axis 314.

[0143] The first flexible container 124 may include a first material in some implementations and the second flexible container 126 may include a second material. The first material may be the same or different from the first material. The first material may be stiffer than the second material and may be resistant to air permeability. The first flexible container 124 may prevent or inhibit the gas used to pressurize the second flexible container 126 from passing through the first flexible container 124 and accessing the reagent 118 within the first flexible container 124. The first flexible container 124 may be metalized to deter the gas from passing into the interior 132 of the first flexible container 124, for example.

[0144] The coupling 128 has a pressure fluidic line 316 and a reagent fluidic line 318. The pressure fluidic line 316 is shown extending between the pressure port 144 and the second portion 140 of the coupling 128 and the reagent fluidic line 318 is shown extending between the reagent port 142 and the first portion 138 of the coupling 128. The pressure fluidic line 316 extends through the second portion 140 of the coupling 128 to enable the pressure source 106 of the system 100 of FIG. 1 to change the pressure within the second flexible container 126 and the reagent fluidic line 318 extends through the first portion 138 and the second portion 140 of the coupling 128 to enable the reagent 118 to flow out of the first flexible container 124.

[0145] FIG. 4 is an isometric view of the coupling 128 of the reagent reservoir 300 of FIG. 2. The coupling 128 is a fitment 320 and may be referred to as a dual fitment because the fitment 320 includes both the first portion 138 and the second portion 140. The coupling also has spacers 322 that are positioned between the first portion 138 and the second portion 140. The spacers 322 define a gap 324 between the first portion 138 and the second portion 140 and allow both of the couplings between the first flexible container 124 and the first portion 138 and between the second flexible container 126 and the second portion 140 to not affect one another.

[0146] The first portion 138 of the coupling 128 has opposing first side walls 326, 328 that form a first canoe shape 330 and the second portion 140 of the coupling 128 has opposing second side walls 332, 334 that form a second canoe shape 336. The first canoe shape 330 is smaller than the second canoe shape 336. The first canoe shape 330 has a smaller width and a smaller depth than the second canoe shape 336 shown. The first flexible container 124 and the second flexible container 126 can each form a coupling with the corresponding canoe shapes 330, 336 as a result of the sizing of the canoe shapes 330, 336, with the first flexible container 124 coupled to the first canoe shape 330 and the second flexible container 126 coupled to the second canoe shape 336. The first canoe shape 330 and the second canoe shape 336 may alternatively be similar or the same sizes. Manufacturing the coupling 128 may be simplified if the canoe shapes 330, 336 are similar or the same. The coupling 128 may be more easily coupled to the first flexible container 124 and the second flexible container 126 if the canoe shapes 330, 336 are similar or the same. [0147] Each of the first portion 138 and the second portion 140 include ribs 338 that are separated by slots 340. The ribs 338 and the slots 340 may facilitate manufacturability of the coupling 128. The coupling 128 may be an injection molded part in some implementations.

[0148] FIG. 5 is a detailed isometric cross-sectional view of the reagent reservoir 300 of FIG. 3. The coupling 128 includes the first portion 138, the second portion 140, the pressure fluidic line 316, and the reagent fluidic line 318. The pressure fluidic line 316 includes an exit port 342 positioned between the first portion 138 and the second portion 140 and defined by the spacer 322. The exit port 342 of the pressure fluidic line 316 is positioned to be fluidly coupled to the second flexible container 126.

[0149] The first portion 138 of the coupling 128 has a first width 344 and the second portion 140 of the coupling 128 has a second width 346 that is wider than the first width 344. The coupling 128 also a head 348 in the implementation shown having the pressure port 144 and the reagent port 142 and a neck 350 positioned between the head 348 and the second portion 140 of the coupling 128.

[0150] FIG. 6 is an isometric view of a plurality of reagent reservoirs 300 of FIG. 3 that are coupled together using a plate 400. The plate 400 defines a plurality of slots 402, 404, 406. The necks 350 of the reagent reservoirs 300 are positioned within the corresponding slots 402, 404, 406 to couple the reagent reservoirs 300 and the plate 400. The reagent receptacle 105 of the system 100 may receive the plate 400 to position and hold the reagent reservoirs 300 within the system 100. The reagent receptacle 105 may include rails and/or define opposing grooves to allow the plate 400 to be received.

[0151] FIG. 7 is a side view of an implementation of a reagent reservoir 500 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 500 includes a flexible container 501 and a coupling 502 coupled to the flexible container 501 . The sipper 154 and the second flexible container 126 are not included in the implementation shown.

[0152] The coupling 502 is similar to the coupling 128 of FIG. 1 . The coupling 502 includes the first portion 138 of the coupling 128 and includes the reagent port 142. The reagent port 142 may be referred to as a port. The reagent port 142 is shown being positioned toward the middle of the coupling 128 and includes a male portion 504 that extends into the interior 132 of the flexible container 501 . The coupling 502 does not include the second portion 140 of the coupling 128 because the second flexible container 126 is omitted, however. [0153] The flexible container 501 includes has the end 130 to which the coupling 502 is coupled and has a tapered bottom 200. The flexible container 501 includes the interior 132 that contains the reagent 1 18. The flexible container 501 has heat seals 212 that include the tapered bottom 200 and form the interior 132. The heat seals 212 are triangular and/or in a triangular arrangement in the plan view shown. The heat seals 212 may be in a different arrangement, however. The heat seals 212 may be pentagonal and/or in a pentagonal arrangement, for example.

[0154] FIG. 8 is a side view of an implementation of a reagent reservoir 550 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 550 is similar to the reagent reservoir 500 of FIG. 7. The reagent reservoir 550 of FIG. 8 includes the sipper 154, however. The sipper 154 of the reagent reservoir 550 of FIG. 8 may be formed by one or more heat seals 208 in some examples. The heat seals 208 may be referred to as heat seal seams. The sipper 154 is shown being coupled to the male portion 504 of the coupling 502. The male portion 504 may be inserted into the channel 210 of the sipper 154 to couple the coupling 502 to the sipper 154. An interference fit may be formed between the male portion 504 and the sipper 154. The sipper 154 of the reagent reservoir 550 may alternatively be implemented by the tube 155.

[0155] FIG. 9 is a side view of an implementation of a reagent reservoir 600 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 600 is similar to the reagent reservoir 550 of FIG. 8.

[0156] The reagent reservoir 600 includes the flexible container 501 including a heat seal seam 602 that forms a baffle 604 within the interior 132. The heat seal seam 602 may be referred to as a first heat seal seam and the baffle 604 may be referred to as a first baffle. A second heat seal seam 606 is shown being included that forms a second baffle 608 within the interior 132. The baffle 604 and the second baffle 608 may form a sub-chamber 609 within the interior 132 of the flexible container 501 .

[0157] The heat seal seams 602, 606 are mirror images of one another across a central axis 610 of the reagent reservoir 650 in the implementation shown. The reagent port 142 is shown being substantially coaxial with the central axis 610. As set forth herein, the phrase “substantially coaxial” means within about 5 degrees of being coaxial including being coaxial itself. The flexible container 501 has a second end 612 and the baffle 604 extends toward the second end 612. The baffle 604 may be positioned at an acute angle relative to the second end 612. The baffle 604 may be in any position, however. [0158] The sipper 154 is shown extending into the interior 132 of the flexible container 501 and between the first baffle 604 and the second baffle 608. The sipper 154 may be an extension 614 which is coupled to the reagent port 142. The sipper 154 and/or the extension may be implemented by the heat seals 208 that form the channel 210 or may be implemented by the tube 155.

[0159] FIG. 10 is a side view of an implementation of a reagent reservoir 650 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 650 is similar to the reagent reservoir 550 of FIG. 8. The reagent reservoir 650 includes the flexible container 501 including lateral sides 652, 654 that include reinforcements 666. The reinforcements 666 may include the lateral sides 652, 654 having additional material and/or thicker material.

[0160] The reinforcements 666 of the lateral sides 652, 654 additionally or alternatively may include inserts 658. The inserts 658 may be coupled to flexible container 501 . The flexible container 501 of FIG. 10 may include heat seal seams on one or more sides of the inserts 658 to provide a space in which the inserts 658 are positioned.

[0161] FIG. 11 is a side view of an implementation of a reagent reservoir 700 that can be used to implement the reagent reservoir 102 of FIG. 1 . The reagent reservoir 700 is similar to the reagent reservoir 650 of FIG. 10. The reagent reservoir 700 includes a frame 702 carried by the flexible container 501 . The reinforcements 656 of the lateral sides 652, 654 are implemented by the frame 702 in this example.

[0162] The flexible container 501 includes heat seal seams 704, 706, 708 that define the interior 132 and lateral sides 652, 654. A pocket 710 is formed between the heat seal seams 704, 706, 708. The frame 702 is positioned within the pocket 710. The frame 702 may be made of any material including plastic. The frame 702 may exert a force onto the flexible container 501 in directions generally indicated by arrows 712, 714. The frame 702 enables the flexible container 501 to be substantially taught. The frame 702 is shown being U-shaped 716.

[0163] FIG. 12 illustrates a flow chart for a method of using the reagent reservoirs 102, 300, 500, 550, 600, 650, 700, the first flexible container 124, the flexible container 501 , or any of the other implementations disclosed herein. The order of execution of the blocks may be changed, and/or some of the blocks described may be changed, eliminated, combined and/or subdivided into multiple blocks. [0164] The process 750 of FIG. 12 begins with rehydrating fluid flowing through the reagent port 142 and into the interior 136 of the flexible container 124, 501 containing dried reagent 118 (Block 752). The rehydrating fluid may be jet dispensed into the interior 136 of the flexible container 124, 501 . The flexible container 124, 501 includes a tapered bottom 200. The flexible container 124, 501 may include a first heat seal seam 602 and a second heat seal seam 606 that form the tapered bottom 200. Flowing the rehydrating fluid through the reagent port 142 may include flowing the rehydrating fluid through the reagent port 142 of the coupling 128 having a portion 138 coupled to the end 130 of the flexible container 124, 501. The coupling 128 includes the reagent port 142 that is fluidly coupled to the interior 132 of the flexible container 124, 501. Flowing rehydrating fluid through the reagent port 142 and into the interior 136 of the flexible container 124, 501 may include flowing the rehydrating fluid through a sipper 154 extending into the interior 132 of the flexible container 126. Flowing the rehydrating fluid through the sipper 154 may include flowing the rehydrating fluid through the sipper 154 formed by a pair of heat seals 208. Flowing the rehydrating fluid through the sipper 154 may include flowing rehydrating fluid through the sipper 154 including an extension 614 coupled to the reagent port 142 and extending into the interior 132 of the flexible container. The extension 614 may include a tube 155 having a proximal end 156 coupled to the first portion 138 of the coupling 128 and a distal end 158 disposed within the interior 136 of the flexible container 124, 501.

[0165] A vortex is enabled to form within the interior 132 of the flexible container 124, 501 based on an interaction between the rehydrating fluid and the tapered bottom 200 (Block 754). The dried reagent 118 in the interior 132 is rehydrated using the vortex (Block 756). The system 100 may pump the rehydrating fluid and dried reagent 1 18 into and out of the flexible container 124, 501 to rehydrate and/or mix the dried reagent 118 in some examples.

[0166] Foaming within the interior of the interior 136 of the flexible container 501 is reduced (Block 758). Reducing the foaming may include reducing the foaming using baffles 604, 608. The baffles 604, 608 are formed by heat seal seams 602, 606 in some implementations. The heat seal seams 602, 606 may include a first heat seal seam 602 extending inwardly from a first lateral side 652 of the flexible container 501 and a second heat seal seam 606 extending inwardly from a second lateral side 654 of the flexible container 501 . The rehydrating fluid may flow through a sipper 154 extending into the interior 132 of the flexible container 501 and between the baffles 604, 608.

[0167] The flexible container 124, 501 is encouraged to be substantially taut (Block 760). The flexible container 124, 501 may be encouraged to be substantially taught by reinforcing lateral sides 652, 654 of the flexible container 124, 501 . The reinforcements 656 of the lateral sides 652, 654 may include inserts 658. The reinforcements 656 of the lateral sides 652, 654 may include a frame 702. The frame 702 may be a U-shaped frame 716.

[0168] FIG. 13 illustrates an example of a bag 1300 made in accordance with the present disclosure. The example bag 1300 includes a fitment 1302 and a flexible shell 11304 defining an internal volume 1306 and an external border 1308. In some examples, the bag 1300 could additionally include a sipper (e.g., sipper 1208 of FIG. 24). In the present example, the fitment 1302 forms a watertight seal with the flexible shell 1304 and an inlet/outlet for the internal volume 1306. In various other examples, the bag 1300 could include more or fewer fitments 1302 and/or more or fewer internal volumes 1306. In the present example, the bag 1300 defines a height 131 Oh of approximately 7 centimeters (cm) and a width 1310w of approximately 9 cm. The bag 1300 may be any size, however.

[0169] In the present example, the internal volume 1306 is approximately thirteen milliliters (13 mL). In various other examples, the internal volume 1306 could be larger or smaller as needed. The internal volume 1306 is shaped approximately triangularly, having a first side 1312, second side 1314, and a third side 1316. The first side 1312 is configured to receive the fitment 1302. Additionally, the second and third side 1314, 1316 form a lower vertex 1322 defining an angle 1324. In the present example, the angle 1324 is approximately 60 degrees (°). In various other examples, the angle 1324 could be greater or less than 60° such as approximately 30°, approximately 45°, approximately 75°, approximately 100°, approximately 120°, and so forth. Additionally, although the internal volume 1306 is shown as triangular, in other examples, the shape of the internal volume 1306 could be quadrilateral, pentagonal, hexagonal, heptagonal, and so forth. However, in any of the foregoing alternatives, the lowermost portion of the internal volume 1306 is shown as the lower vertex 1322.

[0170] FIG. 14 illustrates the bag 1300 of FIG. 13 partially filled with dried reagent 1204. The dried reagent 1204 may be lyophilized microspheres. Additionally, the internal volume 1306 is being filled with the rehydration fluid 1206 via the sipper 1208. The rehydration fluid 1206 may be buffer reagent or any other liquid. As shown in FIG. 14, the bag 1300 facilitates improved rehydration of the dried reagent 1204. The bag 1300 improves rehydration by improving vortex formation 1702, causing breaching 1704, and intra-bag microsphere movement 1706.

[0171] As can be seen in FIG. 14, the sipper 1208 causes vortex formation 1702 proximate to the lower vertex 1322. To improve vortex formation 1702, a distal end 1109 of the sipper 1208 is disposed proximate the lower vertex 1322. In the present example, the sipper 1208 passes rehydration fluid 1206 through the sipper 1208 against the second side 1314 and the third side 1316 and causes a natural circular flow of the rehydration fluid 1206. The vortex formation 1702 increases the turbulence of the rehydration fluid 1206, increasing the rehydration contact between the dried reagent 1204 and the rehydration fluid 1206. Additionally, the rehydration fluid 1206 breaches 1704 the dried reagent 1204. The rehydration fluid 1206 pushes up along the angled second and third walls 1314, 1316 and around the dried reagent 1204. The angled second and third walls 1314, 1316, in addition to the rehydration fluid 1206 movement throughout the bag 1300, helps the dried reagent 1204 move within the bag 1300.

[0172] FIG. 15 shows the bag 1300 of FIG. 13 providing higher recovery and better uniformity over other bag designs.

[0173] FIG. 16 illustrates an alternative bag 1900, similar to the bag 1300 of FIG. 13. The bag 1900 includes a fitment 1902 and a flexible shell 1904 defining an internal volume 1906 and an external border 1908. The external border 1908 may be referred to as edges. In some examples, the bag 1900 could additionally include a sipper (e.g., the sipper 1208 of FIG. 24). In the present example, the fitment 1902 forms a watertight seal with the flexible shell 1904 and an inlet/outlet for the internal volume 1906.

[0174] The internal volume 1906 is shaped approximately pentagonally, having a first side 1912; a second side 1914; a third side 1916; a fourth side 1918, disposed between the first and second sides 1912, 1914; and a fifth side 1919, disposed between the first and third sides 1912, 1916. Similar to the bag 1300, the second and third sides 1914, 1916 form a lower vertex 1922 defining an angle 1924. In the present example, the angle 1324 is approximately 90 degrees (°). In various other examples, the angle 1324 could be greater or less than 60° such as approximately 30°, approximately 45°, approximately 75°, approximately 100°, approximately 120°, and so forth.

[0175] The external border 1908 of the bag 1900 are reinforced. In the present example, the external border 1908 are reinforced for example, with a frame 1932 to prevent or deter folding and buckling of the bag 1900. In some examples the frame 1932 is rigid, but could be semi rigid. The frame 1932 may provide rigidity to the external border 1908 of the bag 1900 while maintaining flexibility in the middle of the bag 1900.

[0176] FIGS. 17 and 18 illustrate examples of assembling the bag 11 100, 11200 from an example flexible shell 11202, 1 1202. In the present example, the flexible shell 1 1202 (similar to the flexible shell 1304) is made of a heat sensitive material that can be thermally bonded to itself when sufficient heat and pressure is applied.

[0177] FIG. 17 illustrates the flexible shell 1 1202 having an integrated sipper 11 112, an internal volume 1 11 14, and an external border 1 1116 that include heat seals. For example, the fitment 11122 is sealed to the flexible shell 11202 and the sipper 11 112 is heat sealed to the fitment 11122 and the flexible shell 1 1202. As shown in FIG. 17, the sipper 1 1112 is formed as a heat sealed channel within the internal volume 1 11 14. In some examples, after the fitment 11122 and the sipper 11 112 are sealed, the internal volume 11114 is formed by sealing the flexible shell 11202 in the triangular shape around the sipper 11112. Lastly, the external border 1 1116 can entirely or partially seal around the internal volume 1 1114. Although described as a series of sequential steps, in other examples, the steps may done in any order.

[0178] The integrated sipper 111 12 enables the bag to be manufactured with fewer steps and would reduce or remove the need for a complicated fitment. The integrated sipper may reduce the overall cost of the apparatus by eliminating the integration of a separate sipper as well as reduce manufacturing and assembly complexity overall. As shown in this FIG. 17, one may heat seal/crimp two straight lines in the center of the bag which are very close to each other such that they create a narrow opening at the bottom of the bag. The sipper would benefit from being flush with the fitment to be more effective.

[0179] FIG. 18 illustrates the bag 1 1200 including the sipper 11212, the first internal volume 11214, the second internal volume 1 1215, and the external border 11216 including heat seals. For example, the fitment 1 1222 is sealed to the flexible shell 11202 and the sipper 11212 is connected to the fitment 1 1122 and the flexible shell 1 1202. As shown in FIG. 18, the sipper 11212 may be partially secured in place via baffles 1 1224. In the present example, the baffles 11224 are heat sealed portions of the flexible shell 11202. In some examples, after the fitment 11122 and the sipper 11212 are sealed, the internal volume 1 1214 is formed by sealing the flexible shell 11202 in the triangular shape around the sipper 11212. Lastly, the external border 111 16 can entirely or partially seal around the internal volume 1 1214. Although described as a series of sequential steps, in other examples, the steps may done in any order.

[0180] FIG. 19 illustrates the bag 1300 of FIG. 13 including an external border 1308 spaced apart from the internal volume 1306. The external border 1308 is adapted to receive a rigid frame 1402 (e.g., U-shaped hook). The rigid frame 1402 is inserted along a longitudinal axis 1404 into an open end of the flexible shell 1304. The rigid frame 1402 adds rigidity and structure to the bag 1300.

[0181] FIG. 20 illustrates a cross-sectional view of the bag 1300 of FIG. 13 as the internal volume 1306 is filled with dried reagent 1204 and/or rehydration fluid 1206. In a first state 1502, the bag 1300 defines a first thickness 1504. In a second state 1512, after the bag has been partially or fully filled with dried reagent 1204 and/or rehydration fluid 1206, the bag 1300 defines a second thickness 1514. As shown in FIG. 19, as the internal volume 1306 is filled, the first thickness 1504 increases to the second thickness 1514.

[0182] In various examples, the horizontal expansion of the bag 1300 from the first state 1502 to the second state 1512 can help facilitate breaching (described above). For example, if the first thickness 1504 corresponds to being filled with only a dried reagent 1204, after filling with a rehydration fluid 1206, the expansion of the bag 1300 will reduce the force necessary to breach through the dried reagent 1204.

[0183] FIG. 21 illustrates a cross-sectional view of the bag 1300 of FIG. 13 with a first fitment 1302’ and a second fitment 1302”. As shown, the size of the fitment 1302 (e.g., 1302’, 1302”) can affect the natural state of the bag 1300. For example, a smaller fitment 1302' can cause the bag 1300 to be in an open and undeformed configuration 1602. Alternatively, a larger fitment 1302” can cause a distortion in the bag 1300 and be in a deformed configuration 1612. In some examples, the deformed configuration 1612 better facilitates the rehydration fluid 1206 breaching (as described above). The deformed state may create less uniform flow that promotes breaching as an example.

[0184] FIG. 22 illustrates the improved recovery and uniformity possible with the bags 1300, 1900 as disclosed herein. As used herein, recovery refers to the amount of lyophilized microspheres (e.g., dried reagent 1204) that are dissolved in liquid (e.g., rehydration fluid 1206) measured as a percentage of lyophilized microspheres originally present. As shown in the recovery chart 11002, the bag 1300 has a recovery of approximately seven percent (7%) higher than a tube shaped bag. Similarly, the uniformity chart 1004 shows improved uniformity for the bag 1300 relative to a tube-shaped bag. As used herein, uniformity refers to the homogeneity of the mixture. A lower uniformity value may be preferential. As can be shown in FIG. 19, the bag 1300 provides a lower uniformity value compared to the tube-shaped bag. A lower uniformity value may mean that the mixture is more uniform; in other words, less variable. The lower percentage value indicates that less of the mixture is non-uniform in some examples. [0185] The bag 1300, 1900 may enable consistently high recovery (greater than 98%) and uniformity (less than 0.5% CV). The folding of the bag due to pressure creates channels that help in rehydration. First injection of buffer wets most microspheres, which leads to easier rehydration. In some examples, vacuumed bag helps against tribo-charging by trapping the microspheres tightly. Table 1 below shows recovery and uniformity of certain reinforced vacuum bags according to selected implementations of the current disclosure.

Table 1

[0186] FIG. 23 illustrates a bag 1202 inefficiently rehydrating dried reagent 1204. In some examples, the bag 1202 may additionally include a sipper 1208 for passing the rehydration fluid 1206 into the bag 1202 and beneath the dried reagent 1204. As shown in FIG. 2, the rehydration fluid 1206 fails to rehydrate all of the dried reagent 1204 (in the form of lyophilized microspheres). Specifically, some of the dried reagent 1204 is never exposed to the rehydration fluid 1206 because the dried reagent 1204 prevents the rehydration fluid 1206 from moving throughout the bag 1202 and/or because bubbles in the bag 1202 limit interaction between the rehydration fluid 1206 and the dried reagent 1204. In some examples, vibration or shaking of the bag 1202 is insufficient to overcome the impediments to rehydrating the dried reagent 1204. [0187] In various examples, the dried reagent 1204 and the rehydration fluid 1206 is provided based on predetermined ratios. In some examples, approximately one milliliter of rehydration fluid 1206 could be provided for approximately every two milliliters of dried reagent 1204. But, in various other examples, any ratio of rehydration fluid 1206 to dried reagent 1204 sufficient and/or necessary to rehydrate the dried reagent 1204 could be used. Further, in some examples, the bag 1202 may include a headspace 1212 of empty space within the bag 1202. In some examples, the headspace 1212 could be filled with air, a specific gas mixture (e.g., nitrogen, oxygen), or vacuum.

[0188] Breaching: FIG. 24 illustrates an example of rehydration fluid 1206 breaching the dried reagent 1204. As shown in FIG. 24, the rehydration fluid 1206 is forced to the top of the bag 1202, ensuring the rehydration fluid 1206 can mix with the dried reagent 1204. The breaching of the rehydration fluid 1206 ensures that substantially all or all of the dried reagent 1204 is brought into contact with the rehydration fluid 1206 in some examples. Further, breaching reduces the likelihood of bubbles forming between the rehydration fluid 1206 and the dried reagent 1204.

[0189] Intra-bag lyophilized reagent movement: As described, movement of the dried reagent 1204 within the bag 1202 improves rehydration of the dried reagent. For example, movement of the dried reagent 1204 increases contact between the dried reagent 1204 and the rehydration fluid 1206. Furthermore, movement of the dried reagent 1204 within the bag reduces the likelihood of air bubbles impeding the contact between the rehydration fluid 1206 and the dried reagent 1204.

[0190] Vortex Formation: Additionally, herein, in accordance with the present disclosure, the formation of a vortex in the bag can facilitate improved rehydration of dried reagent 1204. The hydrodynamic forces of the rehydration fluid 1206 flowing in a vortex assists in the rehydration fluid 1206 breaching (as described above) and causing the dried reagent 1204 to move within the bag 1202. The shape of the bag 1202 and the method of pumping the rehydration fluid 1206 into the bag can cooperatively work to form vortices in the bag 1202.

[0191] Advantages of the disclosed implementations include breaching, vortex formation, and aiding lyophilized microsphere movement, among others. Breaching refers to the phenomenon where the jet injected liquid travels past all the lyo and covers it from the top. This allows the air trapped between the microspheres and air released from the microspheres to escape with the liquid and allows proper mixing. Trapped air can cut off contact between the rest of the microspheres and liquid preventing rehydration. The 60° bag angle may be beneficial for liquid to ride along the edges and breach the microsphere top. The 60° bag angle with these size ratios allow for formation of vortexes at the bottom of the bag which increases the homogeneity of mixing. The angled edges allow for the dried reagent to replace the space previously occupied by the now dissolved reagent. This allows for most if not all the lyophilized microspheres to get rehydrated even when the dry fill has more volume than the liquid used for rehydration. Two-part rehydration recipe may be beneficial for rehydrating the dried reagent. Flexibility in the bag material provides benefits as well. Adding microspheres to the bag creates tension in the panels of the bag which helps to reduce or prevent the microspheres from flying around the container and tribo-charging. Also, during rehydration, the tension in the bag panels help drive the microspheres down into the liquid.

[0192] Different aspects of the bags as disclosed herein may be used to improve the rehydration of the dried reagent. Bag size should be close to or the same as the final volume with little headspace. A “V” shape helps in vortex formation for better rehydration/mixing and allows for easier breaching. For ease of use and/or compatibility with certain systems, the top and bottom of the bag may be flat and not have a gusset. Certain implementations provide for a Fitment in the neck of the bag that is as narrow as possible, for example, slightly larger than the sipper. A large fitment may lead to unpredictable distortion and folding of the bag. The edges of the bag can be reinforced with rods which can keep the bag slightly taut and prevent folding. Bag edges can be reinforced with harder material, with or without rods.

[0193] To enable a greater number of bulk lyophilized reagents and to ease the burden on shipment and allowing multirun sequencing (for example, multiple sequencing runs using a single bag of rehydrated reagent), it may be beneficial to rehydrate and store reagents that are sensitive to oxygen (or other gases in the atmosphere). The ability to vacuum the bag before shipping and then rehydrating it under vacuum reduces or even eliminates any oxygen contamination. Vacuumed bags have the ability to enhance rehydration process by helping in mixing and breaching. One concern with microspheres is tribo-charging during shipment as it leads to loss of reagent (i.e., non-rehydrated dried material) and inconsistent concentration post rehydration (for example, loss can be up to 5-8%). The vacuumed bag holds the microsphere substantially in place during shipping thereby reducing tribo-charging effects. In certain implementations, rehydration may be improved where there is low headspace, in other words, the bag volume is close to the final volume of the output. Vacuum is applied after the bag has been filled with microspheres and a sipper has been inserted into the fitment. It can be applied either from a pouch valve or through a valve in the fitment. The edges of the bag are reinforced (for example, with rigid or semi rigid members similar to coffee sticks) to prevent or deter folding and buckling of the bag. These members may provide rigidity to the frame or outside of the bag while maintaining flexibility in the middle.

[0194] Reinforced vacuumed bags may enable consistently high recovery (greater than 98%) and uniformity (less than 0.5% CV). The folding of the bag due to pressure creates channels that help in rehydration. First injection of buffer wets most microspheres, which leads to easier rehydration. Vacuumed bag helps against tribo-charging by trapping the microspheres tightly.

[0195] Some implementations of the bags disclosed have an integrated sipper. A sipper allows for the dispensed liquid to have high velocity which promotes breaching and forms bigger vortexes that help with mixing. Having an integrated sipper also enables the bag to be manufactured with fewer steps and would reduce or remove the need for a complicated fitment. The integrated sipper may reduce the overall cost of the apparatus by eliminating the integration of a separate sipper as well as reduce manufacturing and assembly complexity overall. One may heat seal/crimp two straight lines in the center of the bag which are very close to each other such that they create a narrow opening at the bottom of the bag. The sipper may benefit from being flush with the fitment to be more effective.

[0196] In certain situations and/or systems, it may be helpful to provide a bag to aid in suppressing any foam that forms during or after rehydration. Some reagents may be susceptible to foam when they are rehydrated by jet mixing/back and forth mixing which may occur with a sipper. Foaming may reduce the recovery of microspheres and can also impact the homogeneity of the rehydrated reagent. Foam may also trap microspheres, thereby reducing, delaying, or preventing their rehydration. Furthermore, for reagents susceptible to microbial growth, foam can act as an epicenter. The bag may be configured or manufactured to create baffles or subchambers by heat sealing/crimping baffles along the wall of the bag which have a narrow opening for the sipper to go through. The acute angled baffles may suppress the foam physically back into the bulk reagent and reduce overall foaming.

[0197] Example 1 . An apparatus, comprising: a flexible container comprising an end and a tapered bottom, the flexible container including an interior containing dried reagent; and a coupling having a portion coupled to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container, wherein rehydrating fluid is to flow through the port and into the interior of the flexible container and wherein an interaction between the rehydrating fluid and the tapered bottom is to rehydrate a substantial portion of the dried reagent.

[0198] Example 2. The apparatus of example 1 or any of the foregoing example, wherein the interaction between the rehydrating fluid and the tapered bottom is to cause a vortex within the interior of the flexible container to rehydrate a substantial portion of the dried reagent

[0199] Example 3. The apparatus of any one of the preceding or foregoing examples, wherein the flexible container comprises a heat-sealed pouch.

[0200] Example 4. The apparatus of example 3 or any one of the preceding or foregoing examples, wherein the heat-sealed pouch comprises a first panel and a second panel.

[0201] Example 5. The apparatus of any one of the preceding or foregoing examples, wherein the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

[0202] Example 6. The apparatus of any one of the preceding or foregoing examples, wherein the tapered bottom comprises a conical bottom.

[0203] Example 7. The apparatus of any one of the preceding or foregoing examples, wherein the tapered bottom comprises a vertex.

[0204] Example 8. The apparatus of example 7 or any one of the preceding or foregoing examples, wherein the vertex has an angle that is less than 180 degrees.

[0205] Example 9. The apparatus of example 7, wherein the vertex has an angle about 45 degrees and about 105 degrees.

[0206] Example 10. The apparatus of example 7or any one of the preceding or foregoing examples, wherein the vertex has an angle between about 60 degrees and about 90 degrees.

[0207] Example 11 . The apparatus of example 1 or any one of the preceding or foregoing examples, wherein the flexible container comprises heat seals that comprise the tapered bottom and form the interior that is triangular.

[0208] Example 12. The apparatus of example 1 or any one of the preceding or foregoing examples, wherein the flexible container comprises heat seals that comprise the tapered bottom and form the interior that is pentagonal. [0209] Example 13. The apparatus of any one of the preceding examples, further comprising a sipper and wherein the sipper is integrally formed with the flexible container.

[0210] Example 14. The apparatus of any one of examples 12 - 13 or any one of the preceding or foregoing examples, wherein the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

[0211 ] Example 15. The apparatus of any one of examples 13 - 14 or any one of the preceding or foregoing examples, wherein the tapered bottom comprises a vertex and the sipper includes a distal end disposed proximate the vertex.

[0212] Example 16. The apparatus of any one of the preceding or foregoing examples, further comprising a heat seal seam that forms a baffle within the interior, wherein the flexible container comprises a second end opposite the end and wherein the baffle extends toward the second end.

[0213] Example 17. The apparatus of any one of the preceding or foregoing examples, further comprising a first heat seal seam that forms a first baffle within the interior and a second heat seal seam that forms a second baffle within the interior.

[0214] Example 18. The apparatus of example 17 or any one of the preceding or foregoing examples, wherein the coupling further comprises an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0215] Example 19. The apparatus of example 18 or any one of the preceding or foregoing examples, wherein the extension comprises a tube having a proximal end coupled to the portion of the coupling and a distal end disposed within the interior of the flexible container.

[0216] Example 20. The apparatus of any one of the preceding or foregoing examples, wherein the flexible container comprises lateral sides comprising reinforcements.

[0217] Example 21 . The apparatus of any one of the preceding or foregoing examples, further comprising a frame carried by the flexible container.

[0218] Example 22. The apparatus of example 21 or any one of the preceding or foregoing examples, wherein the flexible container comprises heat seal seams forming the interior and lateral sides, wherein a pocket is formed between the heat seal seams, the frame positioned within the pocket.

[0219] Example 23. The apparatus of any one of examples 21 - 22 or any one of the preceding or foregoing examples, wherein the frame is U-shaped. [0220] Example 24. The apparatus of any one of the preceding or foregoing examples, wherein the portion of the coupling has opposing first side walls that form a canoe shape.

[0221] Example 25. The apparatus of any one of the preceding or foregoing examples, further comprising a second flexible container having an end and defining a second interior, the flexible container positioned within the second interior, wherein the coupling has the portion coupled to the end of the flexible container and a second portion coupled to the end of the second flexible container, the coupling comprising a pressure port fluidly coupled to the second interior of the second flexible container.

[0222] Example 26. The apparatus of example 25 or any one of the preceding or foregoing examples, wherein the reagent coupling is to couple with the port to enable reagent to flow out of the flexible container and the pressure coupling is to enable a pressure source to change a pressure within the second flexible container to urge the reagent to flow out of the flexible container.

[0223] Example 27. The apparatus of any one of the preceding or foregoing examples, wherein the reagent comprises lyophilized microspheres.

[0224] Example 28. The apparatus of any one of the preceding or foregoing examples, wherein the flexible container and the coupling coupled thereto are vacuum sealed.

[0225] Example 29. A method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom, and rehydrating the dried reagent in the interior using the vortex.

[0226] Example 30. The method of any one of examples 29, 48 or any of the preceding or foregoing examples, wherein the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

[0227] Example 31 . The method of any one of examples 29 - 30, 48 or any of the preceding or foregoing examples, wherein flowing rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container. [0228] Example 32. The method of example 31 or any of the preceding or foregoing examples, wherein flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper formed by a pair of heat seal seams.

[0229] Example 33. The method of example 32 or any of the preceding or foregoing examples, wherein flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper comprising an extension coupled to the port and extending into the interior of the flexible container.

[0230] Example 34. The method of example 33 or any of the preceding or foregoing examples, wherein the extension comprises a tube having a proximal end coupled to a portion of a coupling and a distal end disposed within the interior of the flexible container.

[0231] Example 35. The method of any one of examples 29 - 34 or any of the preceding or foregoing examples, further comprising reducing foaming within the interior of the flexible container using baffles.

[0232] Example 36. The method of example 35 or any of the preceding or foregoing examples, wherein the baffles are formed by heat seal seams.

[0233] Example 37. The method of example 36 or any of the preceding or foregoing examples, wherein the heat seal seams comprise a first heat seal seam extending inwardly from a first lateral side of the flexible container and a second heat seal seam extending inwardly from a second lateral side of the flexible container.

[0234] Example 38. The method of any one of examples 35 - 37 or any of the preceding or foregoing examples, wherein flowing the rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container and between the baffles.

[0235] Example 39. A method, comprising: forming a tapered bottom of a flexible container; depositing dried reagent within an interior of the flexible container; and securing a coupling to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container.

[0236] Example 40. The method of example 39 or any of the preceding or foregoing examples, wherein forming the tapered bottom comprises forming a first heat seal seam and a second heat seal seam. [0237] Example 41 . The method of any one of examples 39 - 40 or any of the preceding or foregoing examples, further comprising forming a sipper integrally formed with the flexible container.

[0238] Example 42. The method of example 41 or any of the preceding or foregoing examples, wherein the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

[0239] Example 43. The method of any one of examples 39 - 42 or any of the preceding or foregoing examples, further comprising forming a baffle within the interior.

[0240] Example 44. The method of any one of examples 39 - 43 or any of the preceding or foregoing examples, further comprising forming a first baffle within the interior and a second baffle within the interior.

[0241] Example 45. The method of example 44 or any of the preceding or foregoing examples, further comprising coupling an extension to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0242] Example 46. The method of example 45 or any of the preceding or foregoing examples, further comprising forming an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

[0243] Example 47. The method of anyone of examples 39 - 46 or any of the preceding or foregoing examples, further comprising vacuum sealing the flexible container.

[0244] Example 48. A method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; breaching the dried reagent with the liquid; and rehydrating the dried reagent in the interior.

[0245] Example 49. The method of example 48 or any of the preceding or foregoing examples, wherein breaching the dried reagent comprises the rehydrating fluid interacting with the tapered bottom.

[0246] Example 50. The method of any one of examples 48 - 49 or any of the preceding or foregoing examples, further comprising forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom. [0247] Example 51 . The method of example 51 or any of the preceding or foregoing examples, wherein rehydrating the dried reagent in the interior comprises rehydrating the dried reagent in the interior using the vortex.

[0248] The flowchart and block diagrams in the Figures illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various examples of the present implementation. In this regard, each block in the flowchart or block diagrams can represent a module, segment, or portion of instructions, which comprises one or more executable instructions for implementing the specified logical function(s). In some alternative implementations, the functions noted in the blocks can occur out of the order noted in the Figures. For example, two blocks shown in succession can, in fact, be executed substantially concurrently, or the blocks can sometimes be executed in the reverse order, depending upon the functionality involved. It will also be noted that each block of the block diagrams and/or flowchart illustration, and combinations of blocks in the block diagrams and/or flowchart illustration, can be implemented by special purpose hardware-based systems that perform the specified functions or acts or carry out combinations of special purpose hardware and computer instructions.

[0249] The terminology used herein is for the purpose of describing particular examples only and is not intended to be limiting. 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. It will be further understood that the terms “comprises” and/or “comprising”, when used in this specification, specify the presence of stated features, integers, steps, processes, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, processes, operations, elements, components and/or groups thereof.

[0250] The corresponding structures, materials, acts, and equivalents of all means or step plus function elements in the claims below, if any, are intended to include any structure, material, or act for performing the function in combination with other claimed elements as specifically claimed. The description of one or more examples has been presented for purposes of illustration and description, but is not intended to be exhaustive or limited to in the form disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art. Any example was chosen and described in order to best explain various aspects and the practical application, and to enable others of ordinary skill in the art to understand various examples with various modifications as are suited to the particular use contemplated. [0251] 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 subject matter disclosed herein at least to achieve the benefits as described herein. In particular, all combinations of claims subject matter appearing at the end of this disclosure are contemplated as being part of the subject matter disclosed herein. It should also be appreciated that terminology explicitly employed herein that also may appear in any disclosure incorporated by reference should be accorded a meaning most consistent with the particular concepts disclosed herein.

[0252] This written description uses examples to disclose the subject matter, and also to enable any person skilled in the art to practice the subject matter, including making and using any devices or systems and performing any incorporated methods. The patentable scope of the subject matter is defined by the claims, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal languages of the claims.

[0253] It is to be understood that the above description is intended to be illustrative, and not restrictive. For example, the above-described examples (and/or aspects thereof) may be used in combination with each other. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the various examples without departing from their scope. While the dimensions and types of materials described herein are intended to define the parameters of the various examples, they are by no means limiting and are merely provided by way of example. Many other examples will be apparent to those of skill in the art upon reviewing the above description. The scope of the various examples should, therefore, be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. In the appended claims, the terms “including” and “in which” are used as the plain- English equivalents of the respective terms “comprising” and “wherein.” Moreover, in the following claims, the terms “first,” “second,” and “third,” etc. are used merely as labels, and are not intended to impose numerical requirements on their objects. Forms of term “based on” herein encompass relationships where an element is partially based on as well as relationships where an element is entirely based on. Forms of the term “defined” encompass relationships where an element is partially defined as well as relationships where an element is entirely defined. Further, the limitations of the following claims are not written in means-plus-function format and are not intended to be interpreted based on 35 U.S.C. § 112, sixth paragraph, unless and until such claim limitations expressly use the phrase “means for” followed by a statement of function void of further structure. It is to be understood that not necessarily all such objects or advantages described above may be achieved in accordance with any particular example. Thus, for example, those skilled in the art will recognize that the systems and techniques described herein may be embodied or carried out in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other objects or advantages as may be taught or suggested herein.

[0254] While the subject matter has been described in detail in connection with only a limited number of examples, it should be readily understood that the subject matter is not limited to such disclosed examples. Rather, the subject matter can be modified to incorporate any number of variations, alterations, substitutions or equivalent arrangements not heretofore described, but which are commensurate with the spirit and scope of the subject matter. Additionally, while various examples of the subject matter have been described, it is to be understood that aspects of the disclosure may include only some of the described examples. Also, while some examples are described as having a certain number of elements it will be understood that the subject matter can be practiced with less than or greater than the certain number of elements. Accordingly, the subject matter is not to be seen as limited by the foregoing description, but is only limited by the scope of the appended claims.

[0255] The terms “connect,” “connected,” “contact” “coupled” and/or the like are broadly defined herein to encompass a variety of divergent arrangements and assembly techniques. These arrangements and techniques include, but are not limited to (1 ) the direct joining of one component and another component with no intervening components therebetween (i.e., the components are in direct physical contact); and (2) the joining of one component and another component with one or more components therebetween, provided that the one component being “connected to” or “contacting” or “coupled to” the other component is somehow in operative communication (e.g., electrically, fluidly, physically, optically, etc.) with the other component (notwithstanding the presence of one or more additional components therebetween). It is to be understood that some components that are in direct physical contact with one another may or may not be in electrical contact and/or fluid contact with one another. Moreover, two components that are electrically connected, electrically coupled, optically connected, optically coupled, fluidly connected or fluidly coupled may or may not be in direct physical contact, and one or more other components may be positioned therebetween. [0256] The terms “including” and “comprising”, as used herein, mean the same thing.

[0257] The terms “substantially”, “approximately”, “about”, “relatively”, or other such similar terms that may be used throughout this disclosure, including the claims, are used to describe and account for small fluctuations, such as due to variations in processing, from a reference or parameter. Such small fluctuations include a zero fluctuation from the reference or parameter as well. For example, they can refer to less than or equal to ± 10%, such as less than or equal to ± 5%, such as less than or equal to ± 2%, such as less than or equal to ± 1%, such as less than or equal to ± 0.5%, such as less than or equal to ± 0.2%, such as less than or equal to ± 0.1%, such as less than or equal to ± 0.05%. If used herein, the terms “substantially”, “approximately”, “about”, “relatively,” or other such similar terms may also refer to no fluctuations, that is, ± 0%.

[0258] As used herein, a “flow cell” can include a device having a lid extending over a reaction structure to form a flow channel therebetween that is in communication with a plurality of reaction sites of the reaction structure. In some examples, a detection device, such as an imaging device and/or optics, are separate from the flow cell. In other examples, a flow cell can include a detection device that detects designated reactions that occur at or proximate to the reaction sites. A flow cell may include a solid-state light detection or “imaging” device, such as a Charge- Coupled Device (CCD) or Complementary Metal-Oxide Semiconductor (CMOS) (light) detection device. The CMOS detection device or sensor, for example, may include a plurality of detection pixels that detects incident emission signals. In some examples, each detection pixel corresponds to a reaction site. In other examples, there may be more or fewer pixels than the number of reaction sites. Likewise, a detection pixel in some examples corresponds to a single sensing element to create an output signal. In other examples, a detection pixel corresponds to multiple sensing elements to create an output signal. A flow cell can also or alternatively include two (or more) opposing sensors, without a lid. As one specific example, a flow cell can fluidically, electrically, or both fluidically and electrically couple to a cartridge, which can fluidically, electrically, or both fluidically and electrically couple to a bioassay system. A cartridge and/or bioassay system may deliver a reaction solution to reaction sites of a flow cell according to a predetermined protocol (e.g., sequencing-by-synthesis), and perform a plurality of imaging events. For example, a cartridge and/or bioassay system may direct one or more reaction solutions through the flow channel of the flow cell, and thereby along the reaction sites. At least one of the reaction solutions may include four types of nucleotides having the same or different fluorescent labels. In some examples, the nucleotides bind to the reaction sites of the flow cell, such as to corresponding oligonucleotides at the reaction sites. The cartridge, bioassay system, or the flow cell itself in some examples then illuminates the reaction sites using an excitation light source (e.g., solid-state light sources, such as light-emitting diodes (LEDs)). In some examples, the excitation light has a predetermined wavelength or wavelengths, including a range of wavelengths. The fluorescent labels excited by the incident excitation light may provide emission signals (e.g., light of a wavelength or wavelengths that differ from the excitation light and, potentially, each other) that may be detected by the light sensors of the flow cell.

[0259] Flow cells described herein perform various biological or chemical processes and/or analysis. More specifically, the flow cells described herein may be used in various processes and systems where it is desired to detect an event, property, quality, or characteristic that is indicative of a designated reaction. For example, flow cells described herein may include or be integrated with light detection devices, sensors, including but not limited to, biosensors, and their components, as well as bioassay systems that operate with sensors, including biosensors.

[0260] The flow cells facilitate a plurality of designated reactions that may be detected individually or collectively. The flow cells perform numerous cycles in which the plurality of designated reactions occurs in parallel. For example, the flow cells may be used to sequence a dense array of DNA features through iterative cycles of enzymatic manipulation and light or image detection/acquisition. As such, the flow cells may be in fluidic communication with one or more microfluidic channels that deliver reagents or other reaction components in a reaction solution to a reaction site of the flow cells. The reaction sites may be provided or spaced apart in a predetermined manner, such as in a uniform or repeating pattern. Alternatively, the reaction sites may be randomly distributed. Each of the reaction sites may be associated with one or more light guides and one or more light sensors that detect light from the associated reaction site. In one example, light guides include one or more filters for filtering certain wavelengths of light. The light guides may be, for example, an absorption filter (e.g., an organic absorption filter) such that the filter material absorbs a certain wavelength (or range of wavelengths) and allows at least one predetermined wavelength (or range of wavelengths) to pass therethrough. In some flow cells, the reaction sites may be located in reaction recesses or chambers, which may at least partially compartmentalize the designated reactions therein. Furthermore, the designation reactions may involve or be more easily detected at temperatures other than at ambient temperatures, for example, at elevated temperatures.

[0261] As used herein, a “designated reaction” includes a change in at least one of a chemical, electrical, physical, or optical property (or quality) of a chemical or biological substance of interest, such as an analyte-of-interest. In particular flow cells, a designated reaction is a positive binding event, such as incorporation of a fluorescently labeled biomolecule with an analyte-of- interest, for example. More generally, a designated reaction may be a chemical transformation, chemical change, or chemical interaction. A designated reaction may also be a change in electrical properties. In particular flow cells, a designated reaction includes the incorporation of a fluorescently-labeled molecule with an analyte. The analyte may be an oligonucleotide and the fluorescently-labeled molecule may be a nucleotide. A designated reaction may be detected when an excitation light is directed toward the oligonucleotide having the labeled nucleotide, and the fluorophore emits a detectable fluorescent signal. In another example of flow cells, the detected fluorescence is a result of chemiluminescence or bioluminescence. A designated reaction may also increase fluorescence (or Forster) resonance energy transfer (FRET), for example, by bringing a donor fluorophore in proximity to an acceptor fluorophore, decrease FRET by separating donor and acceptor fluorophores, increase fluorescence by separating a quencher from a fluorophore, or decrease fluorescence by co-locating a quencher and fluorophore. A biological or chemical analysis may include detecting a designated reaction.

[0262] As used herein, “downstream” refers to being situated in a direction where a net volume of fluid flows towards. For example, if the net flow of fluid flows from a first source, to a second source, such that after a relevant period of time, for example after a DNA sequencing run, more fluid flows from the first source to a second source, the second source is downstream from the first source.

[0263] As used herein, “electrically coupled” and “optically coupled” refers to a transfer of electrical energy and light waves, respectively, between any combination of a power source, an electrode, a conductive portion of a substrate, a droplet, a conductive trace, wire, waveguide, nanostructures, other circuit segment and the like. The terms electrically coupled and optically coupled may be utilized in connection with direct or indirect connections and may pass through various intermediaries, such as a fluid intermediary, an air gap and the like. Likewise, “fluidically coupled” refers to a transfer of fluid between any combination of sources. The term fluidically coupled may be utilized in connection with direct or indirect connections, and may pass through various intermediaries, such as channels, wells, pools, pumps, and the like.

[0264] As used herein, a “reaction solution,” “reaction component” or “reactant” includes any substance that may be used to obtain at least one designated reaction. For example, potential reaction components include reagents, enzymes, samples, other biomolecules, and buffer solutions, for example. The reaction components may be delivered to a reaction site in the flow cells disclosed herein in a solution and/or immobilized at a reaction site. The reaction components may interact directly or indirectly with another substance, such as an analyte-of- interest immobilized at a reaction site of the flow cell.

[0265] As used herein, the term “reaction site” is a localized region where at least one designated reaction may occur. A reaction site may include support surfaces of a reaction structure or substrate where a substance may be immobilized thereon. For example, a reaction site may include a surface of a reaction structure (which may be positioned in a channel of a flow cell) that has a reaction component thereon, such as a colony of nucleic acids thereon. In some flow cells, the nucleic acids in the colony have the same sequence, being for example, clonal copies of a single stranded or double stranded template. However, in some flow cells a reaction site may contain only a single nucleic acid molecule, for example, in a single stranded or double stranded form.

[0266] As used herein, the term “transparent” refers to allowing all or substantially all visible and non-visible electromagnetic radiation or light of interest to pass through unobstructed; the term “opaque” refers to reflecting, deflecting, absorbing, or otherwise obstructing all or substantially all visible and non-visible electromagnetic radiation or light of interest from passing through; and the term “non-transparent” refers to allowing some, but not all, visible and non-visible electromagnetic radiation or light of interest to pass through unobstructed

[0267] As used herein, the term “waveguide” refers to a structure that guides waves, such as electromagnetic waves, with minimal loss of energy by restricting the transmission of energy to a particular direction or range of directions.

Claims

CLAIMS What is claimed is:

1. An apparatus, comprising: a flexible container comprising an end and a tapered bottom, the flexible container including an interior containing dried reagent; and a coupling having a portion coupled to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container, wherein rehydrating fluid is to flow through the port and into the interior of the flexible container and wherein an interaction between the rehydrating fluid and the tapered bottom is to rehydrate a substantial portion of the dried reagent.

2. The apparatus of claim 1 , wherein the interaction between the rehydrating fluid and the tapered bottom is to cause a vortex within the interior of the flexible container to rehydrate a substantial portion of the dried reagent

3. The apparatus of any one of the preceding claims, wherein the flexible container comprises a heat-sealed pouch.

4. The apparatus of claim 3, wherein the heat-sealed pouch comprises a first panel and a second panel.

5. The apparatus of any one of the preceding claims, wherein the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

6. The apparatus of any one of the preceding claims, wherein the tapered bottom comprises a conical bottom.

7. The apparatus of any one of the preceding claims, wherein the tapered bottom comprises a vertex.

8. The apparatus of claim 7, wherein the vertex has an angle that is less than 180 degrees.

9. The apparatus of claim 7, wherein the vertex has an angle about 45 degrees and about 105 degrees.

10. The apparatus of claim 7, wherein the vertex has an angle between about 60 degrees and about 90 degrees.

11 . The apparatus of claim 1 , wherein the flexible container comprises heat seals that comprise the tapered bottom and form the interior that is triangular.

12. The apparatus of claim 1 , wherein the flexible container comprises heat seals that comprise the tapered bottom and form the interior that is pentagonal.

13. The apparatus of any one of the preceding claims, further comprising a sipper and wherein the sipper is integrally formed with the flexible container.

14. The apparatus of any one of claims 12 - 13, wherein the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

15. The apparatus of any one of claims 13 - 14, wherein the tapered bottom comprises a vertex and the sipper includes a distal end disposed proximate the vertex.

16. The apparatus of any one of the preceding claims, further comprising a heat seal seam that forms a baffle within the interior, wherein the flexible container comprises a second end opposite the end and wherein the baffle extends toward the second end.

17. The apparatus of any one of the preceding claims, further comprising a first heat seal seam that forms a first baffle within the interior and a second heat seal seam that forms a second baffle within the interior.

18. The apparatus of claim 17, wherein the coupling further comprises an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

19. The apparatus of claim 18, wherein the extension comprises a tube having a proximal end coupled to the portion of the coupling and a distal end disposed within the interior of the flexible container.

20. The apparatus of any one of the preceding claims, wherein the flexible container comprises lateral sides comprising reinforcements.

21 . The apparatus of any one of the preceding claims, further comprising a frame carried by the flexible container.

22. The apparatus of claim 21 , wherein the flexible container comprises heat seal seams forming the interior and lateral sides, wherein a pocket is formed between the heat seal seams, the frame positioned within the pocket.

23. The apparatus of any one of claims 21 - 22, wherein the frame is U-shaped.

24. The apparatus of any one of the preceding claims, wherein the portion of the coupling has opposing first side walls that form a canoe shape.

25. The apparatus of any one of the preceding claims, further comprising a second flexible container having an end and defining a second interior, the flexible container positioned within the second interior, wherein the coupling has the portion coupled to the end of the flexible container and a second portion coupled to the end of the second flexible container, the coupling comprising a pressure port fluidly coupled to the second interior of the second flexible container.

26. The apparatus of claim 25, wherein the reagent coupling is to couple with the port to enable reagent to flow out of the flexible container and the pressure coupling is to enable a pressure source to change a pressure within the second flexible container to urge the reagent to flow out of the flexible container.

27. The apparatus of any one of the preceding claims, wherein the reagent comprises lyophilized microspheres.

28. The apparatus of any one of the preceding claims, wherein the flexible container and the coupling coupled thereto are vacuum sealed.

29. A method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom, and rehydrating the dried reagent in the interior using the vortex.

30. The method of any one of claims 29, 48, wherein the flexible container comprises a first heat seal seam and a second heat seal seam that form the tapered bottom.

31 . The method of any one of claims 29 - 30, 48, wherein flowing rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container.

32. The method of claim 31 , wherein flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper formed by a pair of heat seal seams.

33. The method of claim 32, wherein flowing the rehydrating fluid through the sipper comprises flowing the rehydrating fluid through the sipper comprising an extension coupled to the port and extending into the interior of the flexible container.

34. The method of claim 33, wherein the extension comprises a tube having a proximal end coupled to a portion of a coupling and a distal end disposed within the interior of the flexible container.

35. The method of any one of claims 29 - 34, further comprising reducing foaming within the interior of the flexible container using baffles.

36. The method of claim 35, wherein the baffles are formed by heat seal seams.

37. The method of claim 36, wherein the heat seal seams comprise a first heat seal seam extending inwardly from a first lateral side of the flexible container and a second heat seal seam extending inwardly from a second lateral side of the flexible container.

38. The method of any one of claims 35 - 37, wherein flowing the rehydrating fluid through the port and into the interior of the flexible container comprises flowing the rehydrating fluid through a sipper extending into the interior of the flexible container and between the baffles.

39. A method, comprising: forming a tapered bottom of a flexible container; depositing dried reagent within an interior of the flexible container; and securing a coupling to the end of the flexible container, the coupling comprising a port fluidly coupled to the interior of the flexible container.

40. The method of claim 39, wherein forming the tapered bottom comprises forming a first heat seal seam and a second heat seal seam.

41 . The method of any one of claims 39 - 40, further comprising forming a sipper integrally formed with the flexible container.

42. The method of claim 41 , wherein the flexible container comprises a heat seal that forms a channel that forms the sipper within the interior.

43. The method of any one of claims 39 - 42, further comprising forming a baffle within the interior.

44. The method of any one of claims 39 - 43, further comprising forming a first baffle within the interior and a second baffle within the interior.

45. The method of claim 44, further comprising coupling an extension to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

46. The method of claim 45, further comprising forming an extension coupled to the port and extending into the interior of the flexible container and between the first baffle and the second baffle.

47. The method of anyone of claims 39 - 46, further comprising vacuum sealing the flexible container.

48. A method, comprising: flowing rehydrating fluid through a port and into an interior of a flexible container containing dried reagent, the flexible container comprising a tapered bottom; breaching the dried reagent with the liquid; and rehydrating the dried reagent in the interior.

49. The method of claim 48, wherein breaching the dried reagent comprises the rehydrating fluid interacting with the tapered bottom.

50. The method of any one of claims 48 - 49, further comprising forming a vortex within the interior of the flexible container based on an interaction between the rehydrating fluid and the tapered bottom.

51 . The method of claim 51 , wherein rehydrating the dried reagent in the interior comprises rehydrating the dried reagent in the interior using the vortex.