System and method for isolating wash buffer for reuse

Abstract

The present application relates to a system and method for isolating wash buffer for reuse. A system includes a fluid device, a flow control valve, a first reagent fluid reservoir that can be fluidically connected to the fluid device through the flow control valve, a first fluid buffer reservoir that can be fluidically connected to the fluid device through the flow control valve, and a common fluid buffer source that can be fluidically connected to the fluid device through the flow control valve. The flow control valve allows the following flows, including: (i) flow from the first reagent fluid reservoir to the fluid device, (ii) flow from the common fluid buffer source to the fluid device, (iii) flow from the fluid device to the first fluid buffer reservoir, (iv) flow from the first reagent fluid reservoir to the fluid device, and (v) flow from the first fluid buffer reservoir to the fluid device.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of the filing dates of provisional patent application serial number 62 / 741,812, filed on October 5, 2018, and Netherlands application serial number 2021970, filed on November 9, 2018, under 35 U.S.C. §119(e), the disclosures of which are incorporated herein by reference. Technical Field

[0003] The present application relates to systems and methods for isolating wash buffer for reuse. Background Art

[0004] Various assay protocols for clinical and molecular processes are implemented in a system including a fluid handling device to deliver various types of reagent fluids contained in one or more reagent storage components to a reagent destination, to perform one or more fluid operations, such as mixing, processing, reaction, detection, etc. Typically, after each fluid operation, a fluid buffer solution is introduced by the fluid device to rinse off any unused reagent molecules remaining from the previous fluid operation, thereby ensuring that the reagent fluid used in the next fluid operation is not polluted by residual reagent molecules. In order to have a sufficient amount of fluid buffer to rinse the fluid device after each fluid operation, a system (especially a fluid cartridge) usually accommodates a large amount of fluid buffer. However, accommodating a large amount of fluid buffer may be troublesome, because when pursuing size reduction, the fluid cartridge is limited in terms of space availability. In addition, accommodating a large amount of fluid buffer increases the cost of performing various fluid operations. Summary of the invention

[0005] A simplified summary is presented below in order to provide a basic understanding of some aspects described herein. This summary is not an extensive overview of the claimed subject matter. It is neither intended to identify key or important elements of the claimed subject matter nor to describe its scope. Its sole purpose is to present some concepts in a simplified form as a prelude to a more detailed description that is presented later.

[0006] Aspects of the present disclosure include a method comprising the following steps: (a) moving an aliquot of a first reagent fluid into a fluid device; (b) after step (a), moving a volume of a fluid buffer into the fluid device; (c) after step (b), moving at least a portion of the volume of fluid buffer moved in step (b) into a first fluid buffer reservoir; (d) after step (c), moving an aliquot of a second reagent fluid into the fluid device; (e) after step (d), moving a volume of fluid buffer into the fluid device; and (f) after step (e), moving at least a portion of the volume of fluid buffer moved in step (e) into a second fluid buffer reservoir.

[0007] Aspects of the present disclosure include a system comprising: a fluidic device, a flow control valve, a first reagent fluid reservoir capable of being fluidically connected to the fluidic device via the flow control valve, a first fluid buffer reservoir capable of being fluidically connected to the fluidic device via the flow control valve, and a common fluid buffer source capable of being fluidically connected to the fluidic device via the flow control valve. In some examples, the flow control valve allows the following flows, including: (i) flow from the first reagent fluid reservoir to the fluidic device, (ii) flow from the common fluid buffer source to the fluidic device, (iii) flow from the fluidic device to the first fluid buffer reservoir, (iv) flow from the first reagent fluid reservoir to the fluidic device, and (v) flow from the first fluid buffer reservoir to the fluidic device.

[0008] Aspects of the present disclosure include a computer-readable medium encoded with computer-executable instructions that, when executed by a computer controller of an automated system, cause the system to perform the following system processes: (a) moving an aliquot of a first reagent fluid into a fluidic device; (b) following process (a), moving a volume of a fluid buffer into the fluidic device; (c) following process (b), moving at least a portion of the volume of fluid buffer moved in process (b) into a first fluid buffer reservoir; (d) following process (c), moving an aliquot of a second reagent fluid into the fluidic device; (e) following process (d), moving a volume of fluid buffer into the fluidic device; and (f) following process (e), moving at least a portion of the volume of fluid buffer moved in process (e) into a second fluid buffer reservoir.

[0009] This application also provides the following aspects:

[0010] 1). A method comprising:

[0011] (a) moving an aliquot of a first reagent fluid into a fluidic device;

[0012] (b) after step (a), moving a volume of fluid buffer into the fluidic device;

[0013] (c) after step (b), moving at least a portion of the volume of fluid buffer moved in step (b) into a first fluid buffer reservoir;

[0014] (d) after step (c), moving an aliquot of a second reagent fluid into the fluidic device;

[0015] (e) after step (d), moving a volume of fluid buffer into the fluidic device;

[0016] (f) after step (e), moving at least a portion of the volume of fluid buffer moved in step (e) into a second fluid buffer reservoir;

[0017] (g) after step (f), moving an aliquot of the first reagent fluid into the fluidic device; and

[0018] (h) after step (g), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in step (h) is moved from the first fluid buffer reservoir.

[0019] 2) The method according to 1) further includes:

[0020] (i) after step (h), moving an aliquot of the second reagent fluid into the fluidic device; and

[0021] (j) after step (i), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in step (j) is moved from the second fluid buffer reservoir.

[0022] 3). The method according to 2) further comprises:

[0023] (k) after step (h) and before step (i), moving at least a portion of the volume moved in step (h) back into the first fluid buffer reservoir; and

[0024] (1) After step (j), moving at least a portion of the volume moved in step (j) back into the second fluid buffer reservoir.

[0025] 4). The method according to any one of 1) to 3), wherein step (b) comprises moving a certain volume of fluid buffer from the first fluid buffer reservoir to the fluid device, and thereafter, moving a certain volume of fluid buffer from a common fluid buffer reservoir to the fluid device, and

[0026] Wherein step (e) comprises moving a volume of fluid buffer from the second fluid buffer reservoir into the fluidic device, and thereafter, moving a volume of fluid buffer from the common fluid buffer reservoir into the fluidic device.

[0027] 5). The method according to 4), wherein in step (b), the volume of the fluid buffer moved from the first fluid buffer reservoir is preferably substantially equal to the volume of the fluid buffer moved from the common fluid buffer reservoir.

[0028] 6). The method according to any one of 1) to 5), wherein step (b) comprises moving a volume of fluid buffer from the first fluid buffer reservoir to the fluid device; and

[0029] Wherein step (e) comprises moving a volume of fluid buffer from the second fluid buffer reservoir into the fluidic device.

[0030] 7). The method according to any one of 1) to 6), wherein the portion of the fluid buffer moved to the first fluid buffer reservoir in step (c) ranges from approximately 30% to 70% of the volume of the fluid buffer moved in step (b).

[0031] 8). The method according to 2), wherein in step (h), the portion of the fluid buffer moved from the first fluid buffer reservoir ranges from about 30% to 70% of the volume of the fluid buffer moved into the fluid device.

[0032] 9). The method according to any one of 1) to 8), wherein in step (d), the aliquot of the second reagent is not mixed with the aliquot of the first reagent in the fluidic device.

[0033] 10). A system comprising:

[0034] Fluid equipment;

[0035] Flow control valve;

[0036] a first reagent fluid reservoir fluidly connectable to the fluidic device via the flow control valve;

[0037] a first fluid buffer reservoir fluidly connectable to the fluidic device via the flow control valve;

[0038] a common fluid buffer source fluidly connectable to the fluidic device through the flow control valve; and

[0039] a pump operably associated with the fluidic device, the first reagent fluid reservoir, the common buffer source, and the first fluid buffer reservoir;

[0040] wherein the flow control valve and the pump are configured to sequentially cause:

[0041] (i) flow from the first reagent fluid reservoir to the fluidic device,

[0042] (ii) flow from the common fluid buffer source to the fluidic device,

[0043] (iii) flow from the fluidic device to the first fluid buffer reservoir,

[0044] (iv) flow from the first reagent fluid reservoir to the fluidic device, and

[0045] (v) Flow from the first fluid buffer reservoir to the fluidic device.

[0046] 11). The system according to 10), further comprising:

[0047] a second reagent fluid reservoir fluidly connectable to the fluidic device via the flow control valve; and

[0048] a second fluid buffer reservoir fluidly connectable to the fluidic device via the flow control valve;

[0049] wherein the flow control valve and the pump are further configured to cause, after (i)-(iii) and before (iv)-(v):

[0050] (vi) flow from the second reagent fluid reservoir to the fluidic device,

[0051] (vii) flow from the second fluid buffer reservoir to the fluidic device,

[0052] (viii) flow from the common buffer source to the fluidic device, and

[0053] (ix) Flow from the fluidic device to the second fluid buffer reservoir.

[0054] 12). The system according to 10) or 11), wherein the first fluid buffer reservoir comprises a storage channel comprising a consistent cross-sectional dimension across its length.

[0055] 13). A system according to 10), 11) or 12), wherein the first fluid buffer reservoir is fluidically connected to the fluid device via a channel, and the fluid buffer reservoir comprises a storage reservoir comprising a cross-sectional dimension larger than a cross-sectional dimension of the channel.

[0056] 14). A system according to any one of 10)-13), wherein the pump is a syringe pump, so that the syringe pump generates a pressure difference between the fluid device and any one of the first reagent fluid reservoir, the common fluid buffer source or the first fluid buffer reservoir to drive fluid flow.

[0057] 15). The system according to any one of 10) to 14), wherein the flow control valve is a rotary valve.

[0058] 16). The system according to any one of 10)-15) further includes a controller in electrical communication with the flow control valve, wherein the controller is used to transmit commands for controlling the operation of the flow control valve to execute the sequence.

[0059] 17). A system according to any one of 10)-16), wherein the first fluid buffer reservoir contains a first volume of fluid, the first volume being at least 30% of the volume of the fluid contained by the fluidic device.

[0060] 18) A computer readable medium encoded with computer executable instructions, which when executed by a computer controller of an automated system causes the system to perform the following system process:

[0061] (a) moving an aliquot of a first reagent fluid into a fluidic device;

[0062] (b) after process (a), moving a volume of fluid buffer into the fluidic device;

[0063] (c) after process (b), moving at least a portion of the volume of fluid buffer moved in process (b) into a first fluid buffer reservoir;

[0064] (d) after process (c), moving an aliquot of a second reagent fluid into the fluidic device;

[0065] (e) after process (d), moving a volume of fluid buffer into the fluidic device;

[0066] (f) after process (e), moving at least a portion of the volume of fluid buffer moved in process (e) to a second fluid buffer reservoir;

[0067] (g) after process (f), moving an aliquot of the first reagent fluid into the fluidic device; and

[0068] (h) after process (g), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in process (h) is moved from the first fluid buffer reservoir.

[0069] 19). The computer-readable medium according to 18), wherein the system process further comprises:

[0070] (i) after process (h), moving an aliquot of the second reagent fluid into the fluidic device; and

[0071] (j) after process (i), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in process (j) is moved from the second fluid buffer reservoir.

[0072] 20). A computer-readable medium according to 18) or 19), wherein process (b) comprises moving a volume of fluid buffer from the first fluid buffer reservoir to the fluid device, and thereafter, moving a volume of fluid buffer from a common fluid buffer reservoir to the fluid device, and

[0073] Wherein process (e) comprises moving a certain volume of fluid buffer from the second fluid buffer reservoir into the fluid device, and thereafter, moving a certain volume of fluid buffer from the common fluid buffer reservoir into the fluid device.

[0074] Other features and characteristics of the subject matter of the present disclosure, as well as the method of operation, the function of the related elements of structure and the combination of parts and the economy of manufacturing will become more apparent when considering the following description and appended claims, with reference to the accompanying drawings, all of which form a part of this specification, in which like reference numerals indicate corresponding parts in the various figures. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] The accompanying drawings, which are incorporated herein and form a part of the specification, illustrate various examples of the subject matter of the present disclosure. In the drawings, like reference numerals indicate identical or functionally similar elements.

[0076] Figure 1is a schematic diagram of an exemplary system for sequestering and reusing two or more reagent fluids and fluid buffers passing through a fluidic device.

[0077] Figure 2 is a schematic diagram of the system performing the first reagent operation.

[0078] Figure 3 is a schematic diagram of the system performing the first reagent reversal.

[0079] Figure 4 Schematic diagram of the system performing the first step of the first washing operation.

[0080] Figure 5 is a schematic diagram of the system performing the second step of the first washing operation.

[0081] Figure 6 is a schematic diagram of the system performing the first buffer inversion.

[0082] Figure 7 is a schematic diagram of the system performing the second reagent operation.

[0083] Figure 8 is a schematic diagram of the system performing the second reagent reversal.

[0084] Fig. 9 is a schematic diagram of the system performing the first step of the second washing operation.

[0085] Fig.10 is a schematic diagram of the system performing the second step of the second washing operation.

[0086] Fig.11 is a schematic diagram of the system performing the second buffer reversal.

[0087] Fig.12 is a schematic diagram of the system performing a third reagent operation.

[0088] Fig.13 is a schematic diagram of the system performing a third reagent reversal.

[0089] Fig.14 Schematic diagram of the system performing the first step of the third washing operation.

[0090] Fig.15 Schematic diagram of the system performing the second step of the third washing operation.

[0091] Fig.16 is a schematic diagram of the system performing the third buffer inversion.

[0092] Fig.17 is a schematic diagram of the system performing the fourth reagent operation.

[0093] Fig.18 is a schematic diagram of the system performing the fourth reagent reversal.

[0094] Fig.19 Schematic diagram of the system performing the fourth washing operation.

[0095] Fig. 20 is a flow chart of an exemplary method for isolating and reusing two or more reagent fluids and fluid buffers passing through a fluidic device.

[0096] Fig.21 is a schematic diagram of a system including a fluid cartridge incorporated into a processing instrument. DETAILED DESCRIPTION

[0097] Although aspects of the subject matter of the present disclosure may be embodied in various forms, the following description and drawings are intended to disclose only some of these forms as specific examples of the subject matter. Therefore, the subject matter of the present disclosure is not intended to be limited to the forms or examples described and shown.

[0098] Unless otherwise specified, all technical terms, symbols and other technical terms or expressions used in this article have the same meanings as those generally understood by those of ordinary skill in the field to which this disclosure belongs. All patents, applications, published applications and other publications mentioned herein are fully incorporated by reference. If the definition set forth in this section is contrary to or otherwise inconsistent with the definition set forth in the patents, applications, published applications and other publications incorporated herein by reference, then the definition set forth in this section takes precedence over the definition incorporated herein by reference.

[0099] As used herein, "a" or "an" means "at least one" or "one or more" unless indicated otherwise or the context suggests otherwise.

[0100] The description may use relative spatial and / or directional terms when describing the position and / or orientation of a component, device, location, feature, or portion thereof. Unless otherwise specified or otherwise dictated by the context of the description, such terms (including without limitation top, bottom, above, below, under, on top of, upper, lower, to the left, to the right, in front of, behind, beside, adjacent, between, horizontal, vertical, diagonal, longitudinal, transverse, radial, axial, etc.) are used for convenience in referring to such components, devices, locations, features, or portions thereof in the accompanying drawings and are not intended to be limiting.

[0101] Furthermore, unless otherwise noted, any specific dimensions mentioned in this description are merely representative of exemplary embodiments of devices embodying aspects of the present disclosure and are not intended to be limiting.

[0102] The use of the term "approximately" applies to all numerical values ​​specified herein, whether or not explicitly indicated. The term generally refers to a numerical range that a person of ordinary skill in the art considers to be a reasonable amount of deviation from the listed numerical values ​​(i.e., having equivalent functions or results) in the context of the present disclosure. For example, and not intended to be limiting, the term may be interpreted as including a deviation of ±10% of a given numerical value, assuming that such deviation does not change the final function or result of the value. Therefore, in some cases, as will be appreciated by a person of ordinary skill in the art, a value of about 1% may be interpreted as a range from 0.9% to 1.1%.

[0103] As used herein, the term "adjacent" refers to proximity or abutment. Adjacent objects may be spaced apart from one another, or may be in actual or direct contact with one another. In some examples, adjacent objects may be coupled to one another or may be integrally formed with one another.

[0104] As used herein, the terms "substantially" and "substantially" refer to a considerable degree or extent. When used in conjunction with, for example, an event, circumstance, characteristic, or attribute, the terms may refer to instances where the event, circumstance, characteristic, or attribute occurs exactly as well as instances where the event, circumstance, characteristic, or attribute occurs very approximately (e.g., taking into account the general tolerance level or variability of the examples described herein).

[0105] As used herein, the terms "optional" and "optionally" mean that the subsequently described parts, structures, elements, events, environments, characteristics, attributes, etc. may or may not be included or appear, and that the description includes instances in which the parts, structures, elements, events, environments, characteristics, attributes, etc. are included or appear and instances in which they are not included or do not appear.

[0106] As used herein, "reagent" refers to any substance or combination of substances that participate in a molecular assay, other than sample material and the products of the assay. Exemplary reagents include nucleotides, enzymes, amplification oligomers, probes, and salts.

[0107] The term "buffer" as used herein refers to any solution with a controlled pH that can be used to dissolve solid (e.g., lyophilized) substances (e.g., reagents, samples, or a combination thereof) or as a diluent for diluting a liquid (e.g., a liquid reagent, a liquid sample, or a combination thereof; or a solution of a reagent, a sample, or a combination thereof).

[0108] According to various examples, the components and devices described herein can be used in conjunction with a fluid cartridge that can include one or more fluid processing pathways, the fluid processing pathways including one or more elements, such as channels, branch channels, valves, diverters, vents, ports, access areas, through holes, beads, beads containing reagents, covers, reaction components, any combination thereof, and the like. Any element can be in fluid communication with another element.

[0109] All possible combinations of elements and components described in the specification or recited in the claims are contemplated and considered to be part of the present disclosure. It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in more detail below (provided such concepts are not mutually inconsistent) are contemplated to be part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing in the present disclosure are contemplated to be part of the inventive subject matter disclosed herein.

[0110] In the appended claims, the term "including" is used as the plain-English equivalent of the respective term "comprising." The terms "comprising" and "including" are defined herein as open-ended and include not only the listed elements, but also any additional elements. Moreover, in the appended claims, the terms "first," "second," and "third," etc. are used merely as labels, and are not intended to impose numerical requirements on their objects.

[0111] The term "fluid communication" means direct fluid communication, for example, two regions may be in fluid communication with each other via an unobstructed fluid processing passage connecting the two regions or may be capable of being in fluid communication, for example, when two regions are connected via a fluid processing passage, they may be capable of being in fluid communication with each other, and the fluid processing passage may include a valve arranged therein, wherein fluid communication between the two regions can be established when the valve arranged in the fluid processing passage is activated, for example by dissolving a soluble valve, bursting a burstable valve, or otherwise opening the valve.

[0112] Fluid Systems

[0113] While reusing a fluid buffer after flushing it through a fluidic device reduces the amount of fluid buffer required for a fluid sequence process, the used fluid buffer often includes remnants of active reagents because a sequencing program may instruct a fluid handling device to introduce a fluid buffer into a fluidic device directly after introducing an active reagent through the fluidic device. Thus, mixing the reused fluid buffer with other fluid operations of a sequencing process may further contaminate the fluid buffer supplied to the fluidic device and / or compromise the integrity of the fluid operation by inadvertently introducing residual reagents from a previous fluid operation into the reused fluid buffer.

[0114] Therefore, a need exists for improved fluidic systems and methods that allow fluidic buffers to be isolated and reused during sequencing processes to avoid contamination and reduce the volume of fluidic buffers required to be stored by a fluidic cartridge to perform a fluidic sequencing process.

[0115] According to various examples, the system is configured to isolate and reuse the reagent fluid and fluid buffer guided by the fluid device by including a common fluid buffer reservoir containing fresh, unused fluid buffer and at least one special fluid buffer reservoir dedicated to accommodating the fluid buffer of each different reagent for flushing through the fluid device. The system can also include a flow control valve operably associated with the common fluid buffer and at least one special fluid buffer reservoir, so that any one of the common fluid buffer reservoir and at least one special fluid buffer reservoir is selectively connected to the fluid device during the washing operation. Therefore, by using one or more special fluid buffer reservoirs to isolate and reuse fluid buffer, the system minimizes the volume of the fluid buffer used in the fluid process such as fluid sequencing operation (e.g., synthesis sequencing (SBS) operation), and the synthesis sequencing operation includes a splitting process (cleave process), a removal process (scavenger process), an incorporation process (incorporation process) and a scanning process intervened by one or more washing operations.

[0116] Figure 1-19An example system 100 for isolating and reusing reagent fluids and fluid buffers during two or more fluid operations is shown. In some examples, the system 100 includes a fluidic device 102, an inlet channel 103, a flow control valve 104, an outlet channel 105, a pump 106, a set of fluid reservoirs 110, and a set of connecting channels 130. In some examples, the system 100 is part of a fluid box (not shown) that supports various components of the system 100 (e.g., the fluidic device 102, the flow control valve 104, the pump 106, and the set of fluid reservoirs 110), but one or more components of the system 100 may not be supported on a common fluid box or other support structure.

[0117] like Figure 1 As shown, fluid device 102 (e.g., flow cell) is fluidically connected to flow control valve 104 through inlet channel 103. In one example, fluid device 102 is a flow cell, which includes a first glass layer (not shown) and a second glass layer (not shown) that are fixed together and define one or more channels (not shown) that can be fluidically manipulated and optically detected. In various examples, fluid device 102 may include fluid inlet 107, fluid outlet 108, and one or more fluid channels (not shown) are fluidically connected to fluid inlet and fluid outlet to allow fluid processing (e.g., chemical or biochemical determination or other processes or reactions) to occur. In various examples, fluid device 102 is configured to allow various types of fluids (e.g., reagents, buffers, reaction media) to be introduced into fluid inlet 107 to experience fluid processing in one or more fluid channels. In various examples, fluid device 102 is also configured to allow various types of fluids to be flushed out from one or more fluid channels through fluid outlet 108.

[0118] exist Figure 1-19 In the example shown, the inlet channel 103 fluidly connects the fluid inlet 107 of the fluidic device 102 to the flow control valve 104, and the outlet channel 105 fluidly connects the fluid outlet 108 of the fluidic device 102 to the pump 106. In other examples (not shown), the system 100 may include two or more channels that fluidically connect the fluidic device 102 to the flow control valve 104 and two or more channels that fluidically connect the fluidic device 102 to the pump 106.

[0119] In some examples, the set of fluid reservoirs 110 includes two or more reagent fluid reservoirs 112, 114, 116, and / or 118. Figure 1In the example shown, the two or more reagent fluid reservoirs include a first reagent fluid reservoir 112, a second reagent fluid reservoir 114, a third reagent fluid reservoir 116, and a fourth reagent fluid reservoir 118, but the present disclosure contemplates any number of reagent fluid reservoirs. The different reagent fluid reservoirs 112, 114, 116, and / or 118 of the group of fluid reservoirs 110 can have the same or varying sizes (i.e., volumes), for example, all reagent fluid reservoirs 112, 114, 116, and 118 can have the same volume, all reagent fluid reservoirs 112, 114, 116, and 118 can have different volumes, or a subset of the reagent fluid reservoirs 112, 114, 116, and / or 118 can have the same volume, and a subset of the reagent fluid reservoirs 112, 114, 116, and / or 118 can have different volumes, depending on the desired storage volume of the reagent to be stored in each reagent fluid reservoir.

[0120] In some examples, the first reagent fluid reservoir 112 contains a first reagent fluid 122, the first reagent fluid 122 includes a solvent and a first reagent for a first reagent operation (e.g., splitting). In some examples, the second reagent fluid reservoir 114 contains a second reagent fluid 124, the second reagent fluid 124 includes a solvent and a second reagent for a second reagent operation (e.g., scanning). In some examples, the third reagent fluid reservoir 116 contains a third reagent fluid 126, the third reagent fluid 126 includes a solvent and a third reagent for a third reagent operation (e.g., doping). In some examples, the fourth reagent fluid reservoir 128 contains a fourth reagent fluid 128, the fourth reagent fluid 128 includes a solvent and a fourth reagent for a fourth reagent operation (e.g., clearing).

[0121] In some examples, the set of fluid reservoirs 110 includes two or more fluid buffer reservoirs 111, 113, 115, and / or 117. Figure 1 In the example shown, the two or more fluid buffer reservoirs include a common fluid buffer source or reservoir 111 and at least one dedicated fluid buffer reservoir 113, 115, and / or 117, each of which is associated with one of the reagent fluids 122, 124, 126, and / or 128. Figure 1-19In the example shown, at least one dedicated fluid buffer reservoir includes a first fluid buffer reservoir 113, a second fluid buffer reservoir 115 and / or a third fluid buffer reservoir 117, but the present disclosure contemplates any number of dedicated fluid buffer reservoirs. The different fluid buffer reservoirs 111, 113, 115 and / or 117 of the group of reservoirs 110 can have the same or varying sizes (i.e., volumes), for example, all fluid buffer reservoirs 111, 113, 115 and / or 117 can have the same volume, all fluid buffer reservoirs 111, 113, 115 and / or 117 can have different volumes, or a subset of fluid buffer reservoirs 111, 113, 115 and / or 117 can have the same volume, and a subset of fluid buffer reservoirs 111, 113, 115 and / or 117 can have different volumes, depending on the required storage volume of the reagent to be stored in each reagent fluid reservoir.

[0122] In some examples, each of the dedicated fluid buffer reservoirs 113, 115 and / or 117 holds a volume of fluid that is at least 30% of the volume of the fluid held by the fluidic device 102. In some examples, each of the dedicated fluid buffer reservoirs 113, 115 and / or 117 is a cache channel that includes a consistent cross-sectional dimension across its length. In some examples, each of the dedicated fluid buffer reservoirs 113, 115 and / or 117 is a cache well that includes a cross-sectional dimension larger than the cross-sectional dimension of its associated connecting channel 130. In some examples, the cache well does not include any sharp edges or various topographical features and is configured to minimize bubble nucleation so that the cache well does not accumulate bubbles when the fluid is pushed and pushed out of the reservoir.

[0123] exist Figure 1-19 In the example shown, common fluid buffer reservoir 111 contains common fluid buffer 121 (e.g., a cleaning solution including saline and soap) that is not flushed through fluidic device 102. In various examples, each of dedicated buffer reservoirs 113, 115, and / or 117 contains a segregated fluid buffer that is flushed through fluidic device 102 following a reagent operation (e.g., with one of reagent fluids 122, 124, and / or 126) performed in fluidic device 102, but initially dedicated fluid buffer reservoirs 113, 115, and / or 117 may contain unused fluid buffer before any reagent operation occurs.

[0124] exist Figure 6In the example shown, the first buffer reservoir 113 contains a first fluid buffer 123 that includes a mixture of spent fluid buffer 602 that is flushed through the fluidic device 102 after the first reagent fluid 122 is directed through the fluidic device 102 during a first reagent operation. Fig.11 In the example shown, the second buffer reservoir 115 contains a second fluid buffer 125 that includes a mixture of spent fluid buffer 1102 that is flushed through the fluidic device 102 after the second reagent fluid 124 is directed through the fluidic device 102 during a second reagent operation. Fig.16 In the example shown, third buffer reservoir 117 contains third fluid buffer 127 that includes a mixture of spent fluid buffer 1602 that was flushed through fluidic device 102 after third reagent fluid 126 was directed through fluidic device 102 during a fourth reagent operation.

[0125] In some examples, the system may not include a dedicated fluid buffer reservoir for each reagent. For example, Figure 1-20 While the illustrated systems and processes do not include a dedicated fluid buffer reservoir associated with the fourth reagent fluid 128, other example systems and processes may include a dedicated fluid buffer reservoir associated with each reagent fluid.

[0126] exist Figure 1-19 In the example shown, the group of connecting channels 130 includes a corresponding connecting channel 131 extending from its associated fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118 to the flow control valve 104, so that the flow control valve 104 is fluidly connected to each fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118 of the group of fluid reservoirs 110. In other examples (not shown), the set of connecting channels 130 may include two or more channels to fluidly connect the corresponding fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118 to the flow control valve 104, such as a front connecting channel (e.g., the corresponding connecting channel 131 shown) and a rear connecting channel (not shown), so that the used fluid buffer can be recycled to the rear of the corresponding fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118 opposite to the front connecting channel.

[0127] The flow control valve 104 is constructed and arranged to selectively fluidly connect one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117 or 118 of the set of fluid reservoirs 110 to the inlet channel 103, and thus to the fluidic device 102. According to various examples, the flow control valve 104 includes a rotary valve for selectively connecting to one of the connecting channels 131 of the corresponding fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118.

[0128] exist Figure 1-19 In the example shown, the flow control valve 104 is a rotary valve including a rotating body 150 and a valve selector passage 152. In some examples, the rotating body 150 is configured to rotate between a plurality of angular positions so that the valve selector passage 152 can fluidly connect any one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 to the valve outlet port via the corresponding inlet port of each fluid reservoir. When the rotating body 150 is rotated to an angular position such that the valve selector passage 152 is aligned with one of the inlet ports of the selected fluid reservoir 111, 112, 113, 114, 115, 116, 117, or 118, the fluid can flow from the selected reservoir 111, 112, 113, 114, 115, 116, 117, or 118 through the valve selector passage 152 and into the valve outlet port.

[0129] In other examples (not shown), the flow control valve 104 may include any other type of valve to selectively fluidly connect one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 to the inlet channel 103. In other examples (not shown), the flow control valve 104 may include a valve array, such as a plurality of pinch valves or solenoid valves and a manifold, to selectively fluidly connect one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 to the inlet channel 103.

[0130] In various examples, a pump 106 fluidly connected to the outlet channel 105 is configured to apply a pressure difference between any one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117 or 118 of the first set of fluid reservoirs 110 and the outlet channel 105 to promote fluid flow bidirectionally along the corresponding connecting channel 131 of the group of connecting channels 130, the flow control valve 104, the inlet channel 103, the fluid device 102 and the outlet channel 105. The pump 106 may include a syringe pump having an actuator (not shown) operably associated with a syringe. In various examples, the actuator is configured to move the plunger of the syringe in a first direction to generate a negative pressure difference to draw fluid toward the barrel of the syringe through the fluid device 102 (and possibly into the barrel). The actuator is also configured to move the plunger in a second direction opposite to the first direction to generate a positive pressure differential and expel the fluid away from the syringe toward a selected reservoir 111, 112, 113, 114, 115, 116, 117, or 118 of the set of fluid reservoirs 110 (and possibly expel the fluid from the syringe). Thus, by moving in the second direction to generate a positive pressure differential, the pump 106 is configured to cause the fluid contained in the fluidic device 102 or the outlet channel 105 to be expelled through the inlet channel 103, the flow control valve 104, the selected connecting channel 131, and into one of the selected fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118.

[0131] In other examples (not shown), pump 106 may include any other pressure differential creating mechanism capable of reversing the direction of flow.

[0132] Fluidic sequencing operation of the system

[0133] In various examples, such as Figure 2-19As shown, system 100 isolates and reuses reagent fluids and fluid buffers by: (i) selectively directing reagent fluids from any one of reagent fluid reservoirs 112, 114, 116, and / or 118 to fluidic device 102 to perform a reagent operation; (ii) optionally redirecting at least a portion of spent reagent fluid 302, 802, 1302, and / or 1802 directed through fluidic device 102 to the selected reagent fluid reservoirs 112, 114, 116, and / or 118; 18 for reuse in a subsequent reagent operation; (iii) selectively directing fluid buffer from any one of the fluid buffer reservoirs 111, 113, 115 and / or 117 to the fluid device 102 for a washing operation; and (iv) redirecting at least a portion of the used fluid buffer 602, 1102 and / or 1602 directed through the fluid device 102 back to one of the dedicated fluid buffer reservoirs 113, 115 and / or 117 for reuse in a subsequent washing operation.

[0134] refer to Figure 2 , the system 100 can be configured to perform a first reagent operation such that the flow control valve 104 allows fluid to flow from the selected first reagent fluid reservoir 112 of the set of reservoirs 110 to the fluidic device 102. During the first reagent operation, the flow control valve 104 is configured to connect the first reagent fluid reservoir 112 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first reagent fluid reservoir 112). The pump 106 is operated to draw the first reagent fluid 122 from the first reagent fluid reservoir 112 through the flow control valve 104 and into the fluidic device 102. Figure 2 As shown, an aliquot 202 of the first reagent fluid 122 moves through the corresponding connecting channel 131 , the flow control valve 104 , the inlet channel 103 , the fluidic device 102 , the outlet channel 105 and / or into a chamber of the pump 106 .

[0135] refer to Figure 3 , the system 100 can be configured to perform a first reagent reversal operation such that the pump 106 redirects at least a portion of the aliquot 202 of the first reagent fluid 122 moved in the first reagent operation back into the first reagent fluid reservoir 112. During the first reagent reversal operation, the flow control valve 104 remains configured to connect the fluidic device 102 to the first reagent fluid reservoir 112, and the pump 106 is operated to discharge fluid through the fluidic device 102 in the opposite direction back into the first reagent fluid reservoir 112. Figure 3As shown, at least a portion 302 of the aliquot 202 of the first reagent fluid 122 moved in a first reagent operation is received back into the first reagent fluid reservoir 112 for reuse in one or more subsequent first reagent operations.

[0136] refer to Figure 4 and Figure 5 , the system 100 can be configured to perform a first washing operation such that the flow control valve 104 first allows fluid to flow from a selected first fluid buffer reservoir 113 of the group reservoir 110 to the fluid device 102, and then optionally allows fluid to flow from a common fluid buffer reservoir 111 of the group reservoir 110 to the fluid device 102.

[0137] like Figure 4 As shown, during the first portion of the first wash operation, the flow control valve 104 is configured to connect the first fluid buffer reservoir 113 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first fluid buffer reservoir 113). The pump 106 is operated to draw an aliquot of the first fluid buffer 123 from the first fluid buffer reservoir 113 through the fluidic device 102. Figure 4 As shown, the first volume 402 of the first fluid buffer 123 moves through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or enters the chamber of the pump 106 to flush the first reagent fluid 122 remaining in the fluid device 102. In some embodiments, in an example where the first fluid buffer 123 is previously pumped into the chamber of the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the pump 106, the first volume 402 may include Figure 6 The volume 602 of the reused first fluid buffer is shown in FIG.

[0138] like Figure 5 As shown, in some examples, during the second portion of the first wash operation, after the first volume 402 of the first fluid buffer 123 is flushed through the fluidic device 102, the flow control valve 104 is configured to connect the common fluid buffer reservoir 111 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to a corresponding connection channel 131 associated with the common fluid buffer reservoir 111). The pump 106 is operated to draw an aliquot of the common fluid buffer 121 from the common fluid buffer reservoir 111 through the fluidic device 102. Figure 5As shown, the second volume 502 of the common fluid buffer 121 can move through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or into the chamber of the pump 106. In some embodiments, the second volume 502 can be mixed with the first volume 402 in one or more of the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106. Therefore, if the first volume 402 is included in Figure 6 The volume 602 of the reused first fluid buffer shown and described in the figure, then the inclusion of the common fluid buffer 121 can dilute or otherwise reduce the residual first reagent fluid 122 present in the reused first fluid buffer. In addition, by providing the common fluid buffer 121 after the reused first fluid buffer, the common fluid buffer 121 can be fluidically separated from the reused first fluid buffer further downstream in the fluidic device 102.

[0139] In other examples, Figure 5 The second part of the illustrated first wash operation may be omitted, and only the first fluid buffer 123 for the first wash operation may be drawn from the first fluid buffer reservoir 113 .

[0140] refer to Figure 6 , the system 100 can be configured to perform a first buffer reversal operation such that the flow control valve 104 allows fluid to flow to the first fluid buffer reservoir 113. The pump 106 redirects the used fluid buffer of the third volume 602 including the first fluid buffer 123 and / or the common fluid buffer 121 back to the first fluid buffer reservoir 113. In addition to the first fluid buffer 123 and / or the common fluid buffer 121, the third volume 602 can also include at least a portion of the residual first reagent fluid 122. In some instances, the percentage of the common fluid buffer 121 in the third volume 602 is greater than the percentage of the first fluid buffer 123 and / or the reused first fluid buffer. During the first buffer reversal operation, the flow control valve 104 is configured to connect the fluid device 102 to the first fluid buffer reservoir 113 (e.g., by connecting the valve selector channel 152 to the connecting channel 131 associated with the first fluid buffer reservoir 113), and the pump 106 is operated to move the fluid through the fluid device 102 to the first fluid buffer reservoir 113 in the opposite direction. Figure 6As shown, the third volume 602 of used fluid buffer is received back into the first fluid buffer reservoir 113 for reuse in one or more subsequent first wash operations. In some examples, the third volume 602 of used fluid buffer is less than or equal to the second volume 502 moved from the common fluid buffer reservoir 111, in other examples, the third volume 602 of used fluid buffer is greater than the second volume 502 moved from the common fluid buffer reservoir 111, and in other examples, the third volume 602 of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device 102, i.e., the combined first fluid volume 402 moved from the first fluid buffer reservoir 113 and the second volume 502 moved from the common fluid buffer reservoir 111.

[0141] refer to Figure 7 , the system 100 can be configured to perform a second reagent operation such that the flow control valve 104 allows fluid to flow from a selected second reagent fluid reservoir 114 of the set of reservoirs 110 to the fluidic device 102. During the second reagent operation, the flow control valve 104 is configured to connect the second reagent fluid reservoir 114 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to a corresponding connecting channel 131 associated with the second reagent fluid reservoir 114). The pump 106 is operated to draw the second reagent fluid 124 from the second reagent fluid reservoir 114 through the flow control valve 104 and into the fluidic device 102. Figure 7 As shown, aliquot 702 of second reagent fluid 124 moves through corresponding connecting channel 131 , flow control valve 104 , inlet channel 103 , fluidic device 102 , outlet channel 105 and / or into the chamber of pump 106 .

[0142] refer to Figure 8 , the system 100 can be configured to perform a second reagent reversal operation such that the pump 106 redirects at least a portion of the aliquot 702 of the second reagent fluid 124 moved in the second reagent operation back into the second reagent fluid reservoir 114. During the second reagent reversal operation, the flow control valve 104 remains configured to connect the fluidic device 102 to the second reagent fluid reservoir 114, and the pump 106 is operated to discharge the fluid through the fluidic device 102 in the opposite direction back into the second reagent fluid reservoir 114. Figure 8 As shown, at least a portion 802 of the aliquot 702 of the second reagent fluid 124 moved in the second reagent operation is received back into the second reagent fluid reservoir 114 for reuse in one or more subsequent second reagent operations.

[0143] refer to Fig. 9 and Fig.10, the system 100 can be configured to perform a second washing operation such that the flow control valve 104 first allows fluid to flow from the selected second fluid buffer reservoir 115 of the group reservoir 110 to the fluid device 102, and then optionally allows fluid to flow from the common fluid buffer reservoir 111 of the group reservoir 110 to the fluid device 102.

[0144] like Fig. 9 As shown, during the first portion of the second wash operation, the flow control valve 104 is configured to connect the second fluid buffer reservoir 115 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connecting channel 131 associated with the second buffer reservoir 115). The pump 106 is operated to draw an aliquot of the second fluid buffer 125 from the second fluid buffer reservoir 115 through the fluidic device 102. Fig. 9 As shown, the fourth volume 902 of the second fluid buffer 125 moves through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or into the chamber of the pump 106 to flush the second reagent fluid 124 remaining in the fluid device 102. In some embodiments, in an instance where the second fluid buffer 125 is previously pumped into the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106, the fourth volume 902 may include Fig.11 The volume 1102 of the reused second fluid buffer is shown in .

[0145] like Fig.10 As shown, in some examples, during the second portion of the second wash operation, after the fourth volume 902 of the second fluid buffer 125 is flushed through the fluidic device 102, the flow control valve 104 is configured to connect the common fluid buffer reservoir 111 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to a corresponding connection channel 131 associated with the common fluid buffer reservoir 111). The pump 106 is operated to draw an aliquot of the common fluid buffer 121 from the common fluid buffer reservoir 111 through the fluidic device 102. Fig.10 As shown, the fifth volume 1002 of the common fluid buffer 121 can move through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or into the chamber of the pump 106. In some embodiments, the fifth volume 1002 can be mixed with the fourth volume 902 in one or more of the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106. Therefore, if the fourth volume 902 is included in Fig.11The volume 1102 of the reused second fluid buffer shown and described in the figure, then the common fluid buffer 121 can dilute or otherwise reduce the residual second reagent fluid 124 present in the reused second fluid buffer. In addition, by providing the common fluid buffer 121 after the reused second fluid buffer, the common fluid buffer 121 can be fluidically separated from the reused second fluid buffer further downstream in the fluidic device 102.

[0146] In other examples, Fig.10 The second portion of the second wash operation shown may be omitted, and only the second fluid buffer 125 for the second wash operation may be drawn from the second fluid buffer reservoir 115 .

[0147] refer to Fig.11 , the system 100 can be configured to perform a second buffer reversal operation such that the flow control valve 104 allows fluid to flow to the second fluid buffer reservoir 115. The pump 106 redirects the spent fluid buffer of the sixth volume 1102 including the second fluid buffer 125 and / or the common fluid buffer 121 back to the second fluid buffer reservoir 115. The sixth volume 1102 can also include at least a portion of the residual second reagent fluid 124 in addition to the second fluid buffer 125 and / or the common fluid buffer 121. In some examples, the percentage of the common fluid buffer 121 in the sixth volume 1102 is greater than the percentage of the second fluid buffer 125 and / or the reused second fluid buffer. During the second buffer reversal operation, the flow control valve 104 is configured to connect the fluidic device 102 to the second fluid buffer reservoir 115 (e.g., by connecting the valve selector channel 152 to the connecting channel 131 associated with the second fluid buffer reservoir 115), and the pump 106 is operated to move fluid through the fluidic device 102 to the second fluid buffer reservoir 115 in the opposite direction. Fig.11 As shown, the sixth volume 1102 of used fluid buffer is received back into the second fluid buffer reservoir 115 for reuse in one or more subsequent second wash operations. In some examples, the sixth volume 1102 of used fluid buffer is less than or equal to the fifth volume 1002 moved from the common fluid buffer reservoir 111, in other examples, the sixth volume 1102 of used fluid buffer is greater than the fifth volume 1002 moved from the common fluid buffer reservoir 111, and in other examples, the sixth volume 1102 of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device 102, i.e., the combined fourth volume 902 moved from the second fluid buffer reservoir 115 and the fifth volume 1002 moved from the common fluid buffer reservoir 111.

[0148] refer to Fig.12 , the system 100 can be configured to perform a third reagent operation such that the flow control valve 104 allows fluid to flow from the selected third reagent fluid reservoir 116 of the set of reservoirs 110 to the fluidic device 102. During the third reagent operation, the flow control valve 104 is configured to connect the third reagent fluid reservoir 116 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the third reagent fluid reservoir 116). The pump 106 is operated to draw the third reagent fluid 126 from the third reagent fluid reservoir 116 through the flow control valve 104 and into the fluidic device 102. Fig.12 As shown, an aliquot 1202 of the third reagent fluid 126 moves through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluidic device 102, the outlet channel 105 and / or into the chamber of the pump 106.

[0149] refer to Fig.13 , the system 100 can be configured to perform a third reagent reversal operation such that the pump 106 redirects at least a portion of the aliquot 1202 of the third reagent fluid 126 moved during the third reagent operation back into the third reagent fluid reservoir 116. During the third reagent reversal operation, the flow control valve 104 remains configured to connect the fluidic device 102 to the third reagent fluid reservoir 116, and the pump 106 is operated to discharge fluid through the fluidic device 102 in the opposite direction to the third reagent fluid reservoir 116. Fig.13 As shown, at least a portion 1302 of the aliquot 1202 of the third reagent fluid 126 moved during the third reagent operation is received back into the third reagent fluid reservoir 116 for reuse in one or more subsequent third reagent operations.

[0150] refer to Fig.14 and Fig.15 , the system 100 can be configured to perform a third washing operation such that the flow control valve 104 first allows fluid to flow from the selected third fluid buffer reservoir 117 of the group reservoir 110 to the fluid device 102, and then optionally allows fluid to flow from the common fluid buffer reservoir 111 of the group reservoir 110 to the fluid device 102.

[0151] like Fig.14 As shown, during the first portion of the third wash operation, the flow control valve 104 is configured to connect the third fluid buffer reservoir 117 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the third buffer reservoir 117). The pump 106 is operated to draw an aliquot of the third fluid buffer 127 from the third fluid buffer reservoir 117 through the fluidic device 102. Fig.14As shown, the seventh volume 1402 of the third fluid buffer 127 moves through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or into the chamber of the pump 106 to flush the third reagent fluid 126 remaining in the fluid device 102. In some implementations, in an instance where the third fluid buffer 127 was previously pumped into the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106, the seventh volume 1402 may include Fig.16 The volume 1602 of the reused third fluid buffer is shown.

[0152] like Fig.15 As shown, in some examples, during the second portion of the third wash operation, after the seventh volume 1402 of the third fluid buffer 126 is flushed through the fluidic device 102, the flow control valve 104 is configured to connect the common fluid buffer reservoir 111 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to a corresponding connection channel 131 associated with the common fluid buffer reservoir 111). The pump 106 is operated to draw an aliquot of the common fluid buffer 121 from the common fluid buffer reservoir 111 through the fluidic device 102. Fig.15 As shown, the eighth volume 1502 of the common fluid buffer 121 can move through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or into the chamber of the pump 106. In some implementations, the eighth volume 1502 can be mixed with the seventh volume 1402 in one or more of the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106. Therefore, if the seventh volume 1402 is included in Fig.16 The volume 1602 of the reused third fluid buffer shown and described in the figure, then the inclusion of the common fluid buffer 121 can dilute or otherwise reduce the residual third reagent fluid 126 present in the reused third fluid buffer. In addition, by providing the common fluid buffer 121 after the reused third fluid buffer, the common fluid buffer 121 can be fluidically separated from the reused third fluid buffer further downstream in the fluidic device 102.

[0153] In other examples, Fig.15 The second portion of the second wash operation shown may be omitted, and only the third fluid buffer 127 for the third wash operation may be drawn from the third fluid buffer reservoir 117 .

[0154] refer to Fig.16, the system 100 can be configured to perform a third buffer reversal operation such that the flow control valve 104 allows fluid to flow to the third fluid buffer reservoir 127. The pump 106 redirects the spent fluid buffer of the ninth volume 1602 including the third fluid buffer 127 and / or the common fluid buffer 121 back to the third fluid buffer reservoir 117. The ninth volume 1602 can also include at least a portion of the residual third reagent fluid 126 in addition to the third fluid buffer 127 and / or the common fluid buffer 121. In some examples, the percentage of the common fluid buffer 121 in the ninth volume 1602 is greater than the percentage of the third fluid buffer 127 and / or the reused third fluid buffer. During the third buffer reversal operation, the flow control valve 104 is configured to connect the fluidic device 102 to the third fluid buffer reservoir 117 (e.g., by connecting the valve selector channel 152 to the connection channel 131 associated with the third fluid buffer reservoir 117), and the pump 106 is operated to move fluid through the fluidic device 102 to the third fluid buffer reservoir 117 in the reverse direction. Fig.16 As shown, the ninth volume 1602 of used fluid buffer is received back into the third fluid buffer reservoir 117 for reuse in one or more subsequent third wash operations. In some examples, the ninth volume 1602 of used fluid buffer is less than or equal to the eighth volume 1502 moved from the common fluid buffer reservoir 111, in other examples, the ninth volume 1602 of used fluid buffer is greater than the eighth volume 1502 moved from the common fluid buffer reservoir 111, and in other examples, the ninth volume 1602 of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device 102, i.e., the combined seventh volume 1402 moved from the third fluid buffer reservoir 117 and the eighth volume 1502 moved from the common fluid buffer reservoir 111.

[0155] refer to Fig.17 , the system 100 can be configured to perform a fourth reagent operation such that the flow control valve 104 allows fluid to flow from the selected fourth reagent fluid reservoir 118 of the set of reservoirs 110 to the fluidic device 102. During the fourth reagent operation, the flow control valve 104 is configured to connect the fourth reagent fluid reservoir 118 to the fluidic device 102 (e.g., by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the fourth reagent fluid reservoir 118). The pump 106 is operated to draw the fourth reagent fluid 128 from the fourth reagent fluid reservoir 118 through the flow control valve 104 and into the fluidic device 102. Fig.17As shown, aliquot 1702 of fourth reagent fluid 128 moves through corresponding connecting channel 131 , flow control valve 104 , inlet channel 103 , fluidic device 102 , outlet channel 105 and / or into the chamber of pump 106 .

[0156] refer to Fig.18 , the system 100 can be configured to perform a fourth reagent reversal operation such that the pump 106 redirects at least a portion of the aliquot 1702 of the fourth reagent fluid 128 moved during the fourth reagent operation back into the fourth reagent fluid reservoir 118. During the fourth reagent reversal operation, the flow control valve 104 remains configured to connect the fluidic device 102 to the fourth reagent fluid reservoir 118, and the pump 106 is operated to discharge fluid through the fluidic device 102 in the opposite direction to the fourth reagent fluid reservoir 118. Fig.18 As shown, at least a portion 1802 of the aliquot 1702 of the fourth reagent fluid 128 moved during the fourth reagent operation is received back into the fourth reagent fluid reservoir 118 for reuse in a subsequent fourth reagent operation.

[0157] In some examples, it may be infeasible or impractical to reuse the fluid buffer after the washing operation, for example, if the characteristic of the reagent or other materials that is rinsed in the washing operation makes the risk of taking out from the buffer for rinsing the reagent too large. In one example, the fluid buffer for rinsing the reagent in the fluid equipment 102 after the removal process of the SBS operation is combined with fresh buffer to harm the subsequent process of the SBS operation usually, even if the used fluid buffer of a trace is added to the fresh buffer. In this case, the buffer can be for example moved from the common buffer reservoir 111 by the fluid equipment, but the used buffer does not reverse back to the isolated fluid buffer reservoir thereafter. In some examples, when the used fluid buffer exceeds the first contamination level, the risk of reusing the fluid buffer is determined to be too large, and the first contamination level can be determined by experiment.

[0158] In some examples, system 100 can reuse the fluid buffer from the same designated reservoir in two different washing operations, and these two washing operations follow two different reagent operations.For example, if the characteristic of the reagent or other materials of the first reagent operation is good, it means that the existence of reagent or material does not affect or endanger another reagent operation (for example, the second reagent operation), so the fluid buffer for flushing this reagent or material after the first reagent operation can be reused in the washing operation after the second reagent operation.In some examples, when the used fluid buffer was lower than the second pollution level, the used fluid buffer can be determined to be good, and the second pollution level can be determined by experiment.

[0159] refer to Fig.19, the system 100 can be configured to perform a fourth washing operation such that the flow control valve 104 allows fluid to flow from the selected common fluid buffer reservoir 111 of the set of reservoirs 110 to the fluidic device 102. Fig.19 As shown, a certain volume 1902 of the common fluid buffer 121 moves through the corresponding connecting passage 131, flow control valve 104, inlet channel 103, fluid equipment 102, outlet channel 105 and / or enters the chamber of pump 106 to rinse the fourth reagent fluid 128 remaining in fluid equipment 102. However, system 100 does not include a fluid buffer reservoir for possible reuse for storing and isolating the buffer used in the fourth washing operation. Therefore, the buffer of this volume 1902 moved by fluid equipment during the fourth washing operation is not reversed from the chamber of corresponding connecting passage 131, flow control valve 104, inlet channel 103, fluid equipment 102, outlet channel 105 and / or pump 106. More specifically, the fluid buffer can be rinsed by the chamber of corresponding connecting passage 131, flow control valve 104, inlet channel 103, fluid equipment 102, outlet channel 105 and / or pump 106, and moved to a waste reservoir (not shown).

[0160] Methods for isolating and reusing reagent fluids and fluid buffers

[0161] According to various examples, Fig. 20 A method 2000 for isolating and reusing reagent fluids and fluid buffers is shown.

[0162] Method 2000 includes step 2010: moving an aliquot of a first reagent fluid 122 into a fluidic device 102. In some examples, step 2010 includes setting a flow control valve 104 to allow fluid to flow from a first reagent fluid reservoir 112 to the fluidic device 102. In some examples, step 2010 includes starting a motor to rotate the flow control valve 104 to align and fluidically connect a valve selector channel 152 with a corresponding connection channel 131 associated with the first reagent fluid reservoir 112. In some examples, step 2010 includes using a pump 106 to move the first reagent fluid 122 from the first reagent fluid reservoir 112 to the fluidic device 102. In some examples, step 2010 includes using an actuator to move a plunger of a syringe pump 106 in a first direction to generate a negative pressure differential to move the first reagent fluid 122 through the fluidic device 102.

[0163] The method 2000 may also include step 2015 of moving a volume of a fluid buffer into the fluidic device 102. In some examples, step 2015 includes a first portion that includes moving a volume of a first fluid buffer 123 from a first fluid buffer reservoir 113 into the fluidic device 102. In some examples, the first portion of step 2015 includes setting a flow control valve 104 to allow fluid to flow from the first fluid buffer reservoir 113 to the fluidic device 102, and using a pump 106 to move the first fluid buffer 123 from the first fluid buffer reservoir 113 to the fluidic device 102. In some examples, the first portion of step 2015 includes activating a motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with a corresponding connection channel 131 associated with the first fluid buffer reservoir 113. In some examples, the first portion of step 2015 includes using an actuator to move a plunger of a syringe pump 106 in a first direction to generate a negative pressure differential to move the first fluid buffer 123 through the fluidic device 102.

[0164] In some examples, step 2015 includes a second portion that includes moving a volume of common fluid buffer 121 from common fluid buffer reservoir 111 into fluidic device 102 after moving first fluid buffer 123. In some examples, the second portion of step 2015 includes setting flow control valve 104 to allow fluid to flow from common fluid buffer reservoir 111 to fluidic device 102, and using pump 106 to move common fluid buffer 121 from common fluid buffer reservoir 111 to fluidic device 102. In some examples, the second portion of step 2015 includes activating a motor to rotate flow control valve 104 to align and fluidically connect valve selector channel 152 with a corresponding connection channel 131 associated with common fluid buffer reservoir 111. In some examples, the second portion of step 2015 includes using an actuator to move a plunger of syringe pump 106 in a first direction to generate a negative pressure differential to move common fluid buffer 121 through fluidic device 102.

[0165] In some examples, the volume of the first fluid buffer 123 moved from the first fluid buffer reservoir 113 is substantially equal to the volume of the common fluid buffer 121 moved from the common fluid buffer reservoir 111. In other examples, the volume of the first fluid buffer 123 moved from the first fluid buffer reservoir 113 is greater than the volume of the common fluid buffer 121 moved from the common fluid buffer reservoir 111. In still other examples, the volume of the first fluid buffer 123 moved from the first fluid buffer reservoir 113 is less than the volume of the common fluid buffer 121 moved from the common fluid buffer reservoir 111.

[0166] Method 2000 may also include step 2020 of moving at least a portion of the volume of fluid buffer moved in step 2015 into a first fluid buffer reservoir 113. In some examples, step 2020 includes setting a flow control valve 104 to allow flow from the fluid device 102 to the first fluid buffer reservoir 113. In some examples, step 2020 includes starting a motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with a corresponding connection channel 131 associated with the first fluid buffer reservoir 113. In some examples, step 2020 includes using a pump 106 to discharge the fluid buffer from the fluid device 102 to the first fluid buffer reservoir 113. In some examples, step 2020 includes using an actuator to move a plunger of the syringe pump 106 in a second direction to generate a positive pressure differential to move the fluid buffer to the first fluid buffer reservoir 113. In some examples, the portion of the fluid buffer moved to the first fluid buffer reservoir 113 in step 2020 ranges from approximately 30% to 70% of the total volume of the fluid buffer moved to the fluidic device 102 in step 2015. For example, if 50 μL of the first fluid buffer 123 is moved from the first fluid buffer reservoir 113 and 50 μL of the common fluid buffer 121 is moved from the common fluid buffer reservoir 111, then 30 μL to 70 μL of the combined 100 μL volume may be moved back into the first fluid buffer reservoir 113. In some instances, the volume moved back into the first fluid buffer reservoir 113 may be primarily the common fluid buffer 121 based on moving the first fluid buffer 123 farther downstream through the fluidic system.

[0167] Method 2000 may also include step 2025: moving an aliquot of a second reagent fluid 124 into the fluidic device 102. In some examples, step 2025 includes setting the flow control valve 104 to allow fluid to flow from the second reagent fluid reservoir 114 to the fluidic device 102. In some examples, step 2025 includes starting the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connecting channel 131 associated with the second reagent fluid reservoir 114. In some examples, step 2025 includes using the pump 106 to move the second reagent fluid 124 from the second reagent fluid reservoir 114 to the fluidic device 102. In some examples, step 2025 includes using an actuator to move the plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the second reagent fluid 124 through the fluidic device 102.

[0168] The method 2000 may also include a second step 2030 of moving a volume of a fluid buffer into the fluidic device 102. In some examples, step 2030 includes a first portion that includes moving a volume of a second fluid buffer 125 from a second fluid buffer reservoir 115 into the fluidic device 102. In some examples, the first portion of step 2030 includes setting the flow control valve 104 to allow fluid to flow from the second fluid buffer reservoir 115 to the fluidic device 102, and using the pump 106 to move the second fluid buffer 125 from the second fluid buffer reservoir 115 to the fluidic device 102. In some examples, the first portion of step 2030 includes activating the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connection channel 131 associated with the second fluid buffer reservoir 115. In some examples, the first portion of step 2030 includes using an actuator to move a plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the second fluid buffer 125 through the fluidic device 102.

[0169] In some examples, step 2030 includes a second portion that includes moving a volume of common fluid buffer 121 from the common fluid buffer reservoir 111 into the fluidic device 102 after moving the second fluid buffer 125. In some examples, the second portion of step 2030 includes setting the flow control valve 104 to allow fluid to flow from the common fluid buffer reservoir 111 to the fluidic device 102, and using the pump 106 to draw the volume of common fluid buffer 121 from the common fluid buffer reservoir 111 to the fluidic device 102. In some examples, the second portion of step 2030 includes activating the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connection channel 131 associated with the common fluid buffer reservoir 111. In some examples, the second portion of step 2030 includes using an actuator to move a plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the common fluid buffer 121 through the fluidic device 102.

[0170] Method 2000 may also include step 2035 of moving at least a portion of the volume of fluid buffer moved in step 2030 into a second fluid buffer reservoir 115. In some examples, step 2035 includes setting the flow control valve 104 to allow flow from the fluid device 102 to the second fluid buffer reservoir 115. In some examples, step 2035 includes starting the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connection channel 131 associated with the second fluid buffer reservoir 115. In some examples, step 2035 includes using the pump 106 to discharge the fluid buffer from the fluid device 102 to the second fluid buffer reservoir 115. In some examples, step 2035 includes using an actuator to move the plunger of the syringe pump 106 in a second direction to generate a positive pressure differential to move the fluid buffer to the second fluid buffer reservoir 115. In some examples, the portion of the fluid buffer moved to the second fluid buffer reservoir 115 in step 2035 ranges from approximately 30% to 70% of the total volume of the fluid buffer moved into the fluidic device 102 in step 2030. For example, if 50 μL of the second fluid buffer 125 is moved from the second fluid buffer reservoir 115 and 50 μL of the common fluid buffer 121 is moved from the common fluid buffer reservoir 111, then 30 μL to 70 μL of the combined 100 μL volume may be moved back into the second fluid buffer reservoir 115. In some instances, the volume moved back into the second fluid buffer reservoir 115 may be primarily the common fluid buffer 121 based on moving the second fluid buffer 125 farther downstream through the fluidic system.

[0171] Method 2000 may also include step 2040: reusing the first reagent fluid 122 by moving an aliquot of the first reagent fluid 122 into the fluidic device 102. In some examples, step 2040 includes setting the flow control valve 104 to allow fluid to flow from the first reagent fluid reservoir 112 to the fluidic device 102. In some examples, step 2040 includes starting the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connection channel 131 associated with the first reagent fluid reservoir 112. In some examples, step 2040 includes using the pump 106 to move the first reagent fluid 122 from the first reagent fluid reservoir 112 to the fluidic device 102. In some examples, step 2040 includes using an actuator to move the plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the first reagent fluid 122 through the fluidic device 102.

[0172] The method 2000 may also include a step 2045 of reusing the first fluid buffer by moving a volume of the fluid buffer into the fluidic device 102, wherein at least a portion of the volume of the fluid buffer moved in step 2045 is moved from the first fluid buffer reservoir 113, which now includes the used fluid buffer moved to the first fluid buffer reservoir 113 in step 2020. In some examples, step 2045 includes a first portion that includes setting the flow control valve 104 to allow fluid to flow from the first fluid buffer reservoir 113 to the fluidic device 102. In some examples, the first portion of step 2045 includes setting the flow control valve 104 to allow fluid to flow from the first fluid buffer reservoir 113 to the fluidic device 102, and using the pump 106 to move the first fluid buffer 123 from the first fluid buffer reservoir 113 to the fluidic device 102. In some examples, the first portion of step 2045 includes activating the motor to rotate the flow control valve 104 to align and fluidly connect the valve selector channel 152 with the corresponding connection channel 131 associated with the first fluid buffer reservoir 113. In some examples, the first portion of step 2045 includes using an actuator to move a plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the first fluid buffer 123 through the fluidic device 102.

[0173] In some examples, step 2045 includes a second portion that includes moving a volume of common fluid buffer 121 from common fluid buffer reservoir 111 into fluidic device 102 after moving first fluid buffer 123. In some examples, the second portion of step 2045 includes setting flow control valve 104 to allow fluid to flow from common fluid buffer reservoir 111 to fluidic device 102, and using pump 106 to move common fluid buffer 121 from common fluid buffer reservoir 111 to fluidic device 102. In some examples, the second portion of step 2045 includes activating a motor to rotate flow control valve 104 to align and fluidically connect valve selector channel 152 with a corresponding connection channel 131 associated with common fluid buffer reservoir 111. In some examples, the second portion of step 2045 includes using an actuator to move a plunger of syringe pump 106 in a first direction to generate a negative pressure differential to move common fluid buffer 121 through fluidic device 102. In some examples, the volume of the first fluid buffer 123 moved from the first fluid buffer reservoir 113 is substantially equal to the volume of the common fluid buffer 121 moved from the common fluid buffer reservoir 111 .

[0174] In some examples, if the first fluid buffer 123 is to be further reused after step 2045, the method includes the step of moving at least a portion of the volume of fluid buffer moved in step 2045 back into the first fluid buffer reservoir 113, as in step 2020 above. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2045 back into the first fluid buffer reservoir 113 includes setting the flow control valve 104 to allow flow from the fluidic device 102 to the first fluid buffer reservoir 113. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2045 back into the first fluid buffer reservoir 113 includes activating a motor to rotate the flow control valve 104 to align and fluidly connect the valve selector channel 152 with the corresponding connection channel 131 associated with the first fluid buffer reservoir 113. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2045 back into the first fluid buffer reservoir 113 includes using an actuator to move the plunger of the syringe pump 106 in a second direction to generate a positive pressure differential to move the fluid buffer to the first fluid buffer reservoir 113. In some examples, the portion of the fluid buffer moved from the first fluid buffer reservoir 113 in step 2045 ranges from approximately 30% to 70% of the volume of the fluid buffer moved into the fluidic device 102 in step 2045.

[0175] Method 2000 may also include step 2050: reusing the second reagent fluid 124 by moving an aliquot of the second reagent fluid 124 into the fluidic device 102. In some examples, step 2050 includes setting the flow control valve 104 to allow fluid to flow from the second reagent fluid reservoir 114 to the fluidic device 102. In some examples, step 2050 includes starting the motor to rotate the flow control valve 104 to align and fluidically connect the valve selector channel 152 with the corresponding connecting channel 131 associated with the second reagent fluid reservoir 114. In some examples, step 2050 includes using the pump 106 to aspirate the second reagent fluid 124 from the second reagent fluid reservoir 114 to the fluidic device 102. In some examples, step 2050 includes using an actuator to move the plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the second reagent fluid 124 through the fluidic device 102.

[0176] The method 2000 may also include step 2055 of reusing the second fluid buffer by moving a volume of the fluid buffer into the fluidic device 102, wherein at least a portion of the volume of the fluid buffer moved in step 2055 is moved from the second fluid buffer reservoir 115, which now includes the used fluid buffer moved to the second fluid buffer reservoir 115 in step 2035. In some examples, step 2055 includes a first portion that includes setting the flow control valve 104 to allow fluid to flow from the second fluid buffer reservoir 115 to the fluidic device 102. In some examples, the first portion of step 2055 includes setting the flow control valve 104 to allow fluid to flow from the second fluid buffer reservoir 115 to the fluidic device 102, and using the pump 106 to move the second fluid buffer 125 from the second fluid buffer reservoir 115 to the fluidic device 102. In some examples, the first portion of step 2055 includes activating the motor to rotate the flow control valve 104 to align and fluidly connect the valve selector channel 152 with the corresponding connection channel 131 associated with the second fluid buffer reservoir 115. In some examples, the first portion of step 2055 includes using an actuator to move a plunger of the syringe pump 106 in a first direction to generate a negative pressure differential to move the second fluid buffer 125 through the fluidic device 102.

[0177] In some examples, step 2055 includes a second portion that includes moving a volume of common fluid buffer 121 from common fluid buffer reservoir 111 to fluidic device 102 after moving second fluid buffer 125. In some examples, the second portion of step 2055 includes setting flow control valve 104 to allow fluid to flow from common fluid buffer reservoir 111 to fluidic device 102, and using pump 106 to move common fluid buffer 121 from common fluid buffer reservoir 111 to fluidic device 102. In some examples, the second portion of step 2055 includes activating a motor to rotate flow control valve 104 to align and fluidically connect valve selector channel 152 with a corresponding connection channel 131 associated with common fluid buffer reservoir 111. In some examples, the second portion of step 2055 includes using an actuator to move a plunger of syringe pump 106 in a first direction to generate a negative pressure differential to move common fluid buffer 121 through fluidic device 102. In some examples, the volume of second fluid buffer 125 moved from second fluid buffer reservoir 115 is substantially equal to the volume of common fluid buffer 121 moved from common fluid buffer reservoir 111 .

[0178] In some examples, if the second fluid buffer 125 is further reused after step 2055, the method includes the step of moving at least a portion of the volume of fluid buffer moved in step 2055 back into the second fluid buffer reservoir 115, as in step 2035. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2055 back into the second fluid buffer reservoir 115 includes setting the flow control valve 104 to allow flow from the fluidic device 102 to the second fluid buffer reservoir 115. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2055 back into the second fluid buffer reservoir 115 includes activating a motor to rotate the flow control valve 104 to align and fluidly connect the valve selector channel 152 with a corresponding connection channel 131 associated with the second fluid buffer reservoir 115. In some examples, the step of moving at least a portion of the volume of fluid buffer moved in step 2055 back to the second fluid buffer reservoir 115 includes using an actuator to move the plunger of the injection pump 106 in a second direction to create a positive pressure differential to move the fluid buffer to the second fluid buffer reservoir 115.

[0179] Processing equipment

[0180] like Fig.21 Schematically shown in FIG. 1 , in some examples, the system 100 includes a fluid box 52 that supports various components of the system 100 (e.g., a fluid device 102, a flow control valve 104, a pump 106, and a set of fluid reservoirs 110). The fluid box 52 can be operably mounted in the processing instrument 50. In some examples, the fluid box 52 includes an inlet channel 103, a flow control valve 104, at least a portion of an outlet channel 105, the set of fluid reservoirs 110, and the set of connecting channels 130. The fluid device 52 can be operably coupled to the instrument 50, and the instrument 50 can include one or more actuators (e.g., motors) to control the position flow control valve 104 and the pressure applied by the pump 106 to select and move various reagent fluids and fluid buffers. The instrument 50 can also include a waste outlet (not shown) to dispose of any used reagent fluids or fluid buffers. A controller 140 (which may be part of the instrument 50 or may be a separate or remote computer resource operably connected to the instrument 50) controls the operation of the instrument 50 (e.g., the processing of the fluidic device 102 and the operation of the pump 106) and the operation of the fluid cartridge 52 (e.g., the operation of the flow control valve 104).

[0181] Hardware and Software

[0182] Aspects of the present disclosure are implemented through control and computing hardware components, user-created software, data input components, and data output components. The hardware components include components configured to receive one or more input values, execute one or more algorithms stored on a non-transitory machine-readable medium (e.g., software), and perform other operations. Fig. 20 The algorithm described in (described in) implements the calculation and control module (for example, system controller), such as microprocessor and computer, which provides instructions for manipulating or otherwise acting on input values ​​and outputs one or more output values. This output can be displayed or otherwise indicated to the user for providing information to the user, such as information about the state of the instrument or the process performed thereby, or this output can include input to other processes and / or control algorithms. The data input component includes an element, and the data is input by the element for use by the control and computing hardware component. This data input part can include a position sensor, a motor encoder, and a manual input element, such as a graphical user interface, a keyboard, a touch screen, a microphone, a switch, a manually operated scanner, a voice-activated input part, etc. The data output component can include a hard drive or other storage medium, a graphical user interface, a monitor, a printer, an indicator light, or an audible signal element (such as a buzzer, a speaker, a bell, etc.). The software includes instructions stored on a non-transitory computer-readable medium, which when executed by the control and computing hardware causes the control and computing hardware to perform one or more automated or semi-automated processes.

[0183] In some examples, the apparatus may include a control system including a computer controller 140 (in Figure 1-19 100). Controller 140 may be a control system or computer (e.g., a stand-alone computer) connected to any of the devices of system 100, or may include computer components, such as application specific integrated circuits, integrated with any of the devices of system 100. These computer components may include one or more microprocessors, displays, keyboards (and / or other user input devices), memory components, printers, and / or other devices. Controller 140 may be configured to receive input from a user (e.g., user input) and / or feedback devices (e.g., pressure sensors, flow meters, etc.), and manage the performance of fluid operations of system 100. Controller 140 may include software algorithms to implement processes, such as implementing Fig. 20The process of method 2000 shown enables a user to input user-defined parameters related to fluid processing operations into the fluid device 102 of the system 100, schedule different fluid processing operations on the fluid device 102 of the system 100, and / or enable the controller 140 to perform different steps associated with the fluid processing operations, monitor the performance of the fluid processing operations, and / or output results to the user (on a display, printout, etc.).

[0184] like Figure 1-19 As shown, the controller 140 is in electrical communication with the flow control valve 104, the pump 106 (indicated by the dotted lines) and / or with an intermediate device configured to control the flow control valve 104 and / or the pump 106 (e.g., a stepper motor of the flow control valve 104, a motor of the pump 106, etc.), so that the controller 140 can send instructions to control the control valve 104 and the pump 106 to perform different steps associated with the fluid processing operation (e.g., Figure 2-19 The associated processes and / or Fig. 20 In some examples, the controller 140 is configured to transmit a command to the flow control valve 104 to fluidly connect a selected fluid reservoir of the set of reservoirs 110 to the inlet channel 103 so that the fluid from the selected fluid reservoir can flow through the corresponding connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and / or the chamber of the pump 106. In some examples, the controller 140 is configured to transmit a command to the pump 106 to move in a first direction or a second direction to generate a pressure difference between any fluid reservoir of the set of fluid reservoirs 110 and the outlet channel 105 to drive the fluid to flow toward or away from the pump 106.

[0185] In some examples, the controller 140 is configured to access a computer-readable medium encoded with computer-executable instructions to perform the various processes described herein. In some examples, by executing instructions encoded in the computer-readable medium, the controller 140 causes the system 100 to perform the methods and processes described herein or portions thereof, including: (a) moving an aliquot of a first reagent fluid into a fluidic device; (b) after process (a), moving a volume of a fluid buffer into the fluidic device; (c) after process (b), moving at least a portion of the volume of the fluid buffer moved in process (b) into a first fluid buffer reservoir; (d) after process (c), moving an aliquot of a second reagent fluid into the fluidic device; (e) after process (d), moving a volume of a fluid buffer into the fluidic device; and / or (f) after process (e), moving at least a portion of the volume of the fluid buffer moved in process (e) into a second fluid buffer reservoir.

[0186] It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in more detail below (provided that such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing in this disclosure are contemplated as being part of the inventive subject matter disclosed herein. It should also be appreciated that terms used expressly herein that may also appear in any disclosure incorporated by reference should be given the meaning most consistent with the specific concepts disclosed herein.

[0187] Although the subject matter of the present disclosure has been described and illustrated in considerable detail with reference to certain illustrative examples (including various combinations and subcombinations of features), those skilled in the art will readily recognize other examples and variations and modifications thereof that are included within the scope of the present disclosure. Furthermore, the description of such examples, combinations, and subcombinations is not intended to convey that the claimed subject matter requires features or combinations of features other than those expressly recited in the claims. Accordingly, the scope of the present disclosure is intended to include all modifications and variations that are included within the spirit and scope of the following appended claims.

Claims

1. A method for isolating a wash buffer for reuse, comprising: (a) moving an aliquot of a first reagent fluid into a fluidic device; (b) after step (a), moving a volume of fluid buffer into the fluidic device; (c) after step (b), moving at least a portion of the volume of fluid buffer moved in step (b) and used in the fluidic device back from the fluidic device into a first fluid buffer reservoir; (d) after step (c), moving an aliquot of a second reagent fluid into the fluidic device; (e) after step (d), moving a volume of fluid buffer into the fluidic device; (f) after step (e), moving at least a portion of the volume of fluid buffer moved in step (e) and used in the fluidic device back from the fluidic device into a second fluid buffer reservoir; (g) after step (f), moving an aliquot of the first reagent fluid into the fluidic device; and (h) after step (g), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in step (h) is moved from the first fluid buffer reservoir and used in the fluidic device; wherein step (b) comprises moving a volume of fluid buffer from the first fluid buffer reservoir into the fluidic device, and thereafter, moving a volume of fluid buffer from a common fluid buffer reservoir into the fluidic device, and Wherein step (e) comprises moving a volume of fluid buffer from the second fluid buffer reservoir into the fluidic device, and thereafter, moving a volume of fluid buffer from the common fluid buffer reservoir into the fluidic device.

2. The method according to claim 1, further comprising: (i) after step (h), moving an aliquot of the second reagent fluid into the fluidic device; as well as (j) after step (i), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in step (j) is moved from the second fluid buffer reservoir.

3. The method according to claim 2, further comprising: (k) after step (h) and before step (i), moving at least a portion of the volume moved in step (h) back into the first fluid buffer reservoir; as well as (1) After step (j), moving at least a portion of the volume moved in step (j) back into the second fluid buffer reservoir.

4. The method of claim 1, wherein in step (b), the volume of fluid buffer moved from the first fluid buffer reservoir is substantially equal to the volume of fluid buffer moved from the common fluid buffer reservoir.

5. The method according to any one of claims 1-4, wherein the portion of fluid buffer moved to the first fluid buffer reservoir in step (c) ranges from 30% to 70% of the volume of fluid buffer moved in step (b).

6. The method of claim 2, wherein in step (h), the portion of fluid buffer moved from the first fluid buffer reservoir ranges from 30% to 70% of the volume of fluid buffer moved into the fluidic device.

7. The method of any one of claims 1-6, wherein in step (d), the aliquot of the second reagent fluid is not mixed with the aliquot of the first reagent fluid in the fluidic device.

8. A system for isolating wash buffer for reuse, comprising: Fluid equipment; Flow control valve; a first reagent fluid reservoir fluidly connectable to the fluidic device via the flow control valve; a first fluid buffer reservoir fluidly connectable to the fluidic device through the flow control valve and for containing a fluid buffer that has been used to flush a first reagent through the fluidic device; a common fluid buffer source fluidly connectable to the fluidic device through the flow control valve and adapted to contain fresh, unused fluid buffer; a pump operably associated with the fluidic device, the first reagent fluid reservoir, the common fluid buffer source, and the first fluid buffer reservoir; a second reagent fluid reservoir fluidly connectable to the fluidic device via the flow control valve; as well as a second fluid buffer reservoir fluidly connectable to the fluidic device through the flow control valve and for containing a fluid buffer that has been used to flush a second reagent through the fluidic device; wherein the flow control valve and the pump are configured to sequentially cause: (i) flow from the first reagent fluid reservoir to the fluidic device, (ii) flow from the common fluid buffer source to the fluidic device, (iii) flow from the fluidic device to the first fluid buffer reservoir, (iv) flow from the second reagent fluid reservoir to the fluidic device, (v) flow from said common fluid buffer source to said fluidic device, (vi) flow from the fluidic device to the second fluid buffer reservoir, (vii) flow from the first reagent fluid reservoir to the fluidic device, and (viii) Flow from the first fluid buffer reservoir to the fluidic device.

9. The system of claim 8, wherein the first fluid buffer reservoir comprises a storage channel comprising a consistent cross-sectional dimension across its length.

10. The system of claim 8 or 9, wherein the first fluid buffer reservoir is fluidly connected to the fluidic device via a channel, and the first fluid buffer reservoir comprises a storage reservoir comprising a cross-sectional dimension larger than a cross-sectional dimension of the channel.

11. The system of any one of claims 8-10, wherein the pump is a syringe pump, such that the syringe pump generates a pressure differential between the fluid device and any one of the first reagent fluid reservoir, the common fluid buffer source, or the first fluid buffer reservoir to drive fluid flow.

12. The system of any one of claims 8-11, wherein the flow control valve is a rotary valve.

13. The system of any one of claims 8-12, further comprising a controller in electrical communication with the flow control valve, the controller for transmitting commands to control operation of the flow control valve to perform the sequence.

14. The system of any one of claims 8-13, wherein the first fluid buffer reservoir contains a first volume of fluid, the first volume being at least 30% of the volume of fluid contained by the fluidic device.

15. A computer readable medium encoded with computer executable instructions which, when executed by a computer controller of an automated system, cause the system to perform the following system process: (a) moving an aliquot of a first reagent fluid into a fluidic device; (b) after process (a), moving a volume of fluid buffer into the fluidic device; (c) after process (b), moving at least a portion of the volume of fluid buffer moved in process (b) and used in the fluidic device back from the fluidic device to a first fluid buffer reservoir; (d) after process (c), moving an aliquot of a second reagent fluid into the fluidic device; (e) after process (d), moving a volume of fluid buffer into the fluidic device; (f) after process (e), moving at least a portion of the volume of fluid buffer moved in process (e) and used in the fluidic device back from the fluidic device to a second fluid buffer reservoir; (g) after process (f), moving an aliquot of the first reagent fluid into the fluidic device; and (h) after process (g), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in process (h) is moved from the first fluid buffer reservoir and used in the fluidic device; wherein process (b) comprises moving a volume of fluid buffer from the first fluid buffer reservoir into the fluidic device, and thereafter, moving a volume of fluid buffer from a common fluid buffer reservoir into the fluidic device, and Wherein process (e) comprises moving a certain volume of fluid buffer from the second fluid buffer reservoir into the fluid device, and thereafter, moving a certain volume of fluid buffer from the common fluid buffer reservoir into the fluid device.

16. The computer readable medium of claim 15, wherein the system process further comprises: (i) after process (h), moving an aliquot of the second reagent fluid into the fluidic device; as well as (j) after process (i), moving a volume of fluid buffer into the fluidic device, wherein at least a portion of the volume of fluid buffer moved in process (j) is moved from the second fluid buffer reservoir.

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