RU2800867C2 - System and method for isolation and reuse of buffer solution during washing - Google Patents

Abstract

FIELD: analytical instruments for bionanotechnology.

SUBSTANCE: method for isolating and reusing a wash buffer solution, comprising: (a) moving an aliquot of the first liquid reagent into a fluid device; (b) after step (a) - moving the volume of the buffer solution into the fluid device; (c) after step (b), moving at least a portion of the volume of the buffer solution transferred in step (b) into the reservoir of the first buffer solution; (d) after step (c), transferring an aliquot of the second liquid reactant to the fluid device; (e) after step (d) - transferring the volume of the buffer solution into the fluid device; (f) after step (e), moving at least a portion of the volume of the buffer solution transferred in step (e) into the reservoir of the second buffer solution; (g) after step (f), transferring an aliquot of the first liquid reactant to the fluid device; (h) after step (g), transferring the buffer solution volume to the fluid device, wherein at least a portion of the buffer solution volume transferred in step (h) is transferred from the first buffer solution reservoir.

EFFECT: reduced amount of liquid buffer solution.

20 cl, 21 dwg

Description

The technical field to which the invention belongs

The present invention relates to the field of analytical instruments for bionanotechnology, and more specifically to a system and method for isolating and reusing a wash buffer solution.

State of the art

Protocols for various studies for clinical and molecular processes are implemented in systems that include fluid handling equipment to deliver various types of liquid reagents contained in one or more reagent storage containers to a reagent delivery site for one or more fluid handling. such as mixing, processing, reactions, detection, etc. Typically, after each liquid operation, a liquid buffer solution is passed through the fluid device to flush out any unused reagent molecules remaining from the previous liquid operation and thereby ensure that the liquid reagent used in the next liquid operation is not contaminated with residual liquid molecules. reagent. In order to have a sufficient amount of liquid buffer solution to flush the fluid device after each operation with liquids, large volumes of buffer solution are usually provided in the system, in particular in the liquid cartridge. However, accommodating a large volume of buffer solution can be inconvenient because liquid cartridges are limited in terms of available space in order to reduce their size. Moreover, the placement of large volumes of buffer solution increases the cost of various fluid handling operations.

Disclosure of invention

Below, a brief summary will be presented to provide a basic understanding of some aspects of the described invention. This disclosure is not a detailed overview of the claimed subject matter. It is intended neither to identify key or critical elements of the claimed subject matter nor to delineate the scope of the invention. The sole purpose is to present some of the ideas in a simplified form as an introduction to the more detailed description that will be presented below.

Aspects of the present invention encompass a method comprising: (a) transferring an aliquot of the first liquid reactant into a fluid device; (b) after step (a) - moving the volume of the buffer solution into the fluid device; (c) after step (b), transferring at least a portion of the volume of the buffer solution transferred in step (b) into the reservoir of the first buffer solution; (d) after step (c), transferring an aliquot of the second liquid reactant to the fluid device; (e) after step (d) - moving the volume of the buffer solution into the fluid device; (f) after step (e), transferring at least a portion of the volume of buffer solution transferred in step (e) into the second buffer reservoir.

Aspects of the present invention cover a system comprising: a fluid device; flow control valve; a first liquid reactant reservoir configured to be in fluid communication with the fluid device via a flow control valve; a first buffer solution reservoir configured to be in fluid communication with the fluid device via a flow control valve; a common source of buffer solution, configured to be in fluid communication with the fluid device via a flow control valve. In some examples, the flow control valve allows for the following flows: (i) flow from the first liquid reactant reservoir to the fluid device; (ii) flow from the common buffer source to the fluid device; (iii) flow from the fluid device to the reservoir of the first buffer solution; (iv) flow from the first liquid reactant reservoir to the fluid device; and (v) flow from the first buffer solution reservoir to the fluid device.

Aspects of the present invention include a computer readable medium programmed with computer executable instructions which, when executed by the computer controller of the automated system, causes the system to perform the following processes: (a) transferring an aliquot of the first liquid reagent into a fluidic device; (b) after process (a) - transferring a volume of buffer solution to the fluid device; (c) after process (b) - transferring at least a portion of the volume of buffer solution transferred during process (b) into the reservoir of the first buffer solution; (o!) after process (c) - moving an aliquot of the second liquid reagent into the fluid device; (e) after process (d) - transferring a volume of buffer solution into the fluid device; and (f) after process (e), transferring at least a portion of the volume of buffer solution transferred during process (e) into the second buffer tank.

Other distinguishing features and characteristics of the subject matter of the present invention, methods of operation, functions of the respective structural elements and combination of parts, and manufacturing economics will become more apparent upon reading the following description and the appended claims with reference to the accompanying drawings, which together form a part of the present description, when In this case, in the various figures, the corresponding elements are assigned the same reference designations.

Brief description of the drawings

The accompanying drawings, which are included and form part of the description, illustrate various embodiments of the invention. In the drawings, like reference numbers designate identical or functionally similar elements.

Fig. 1 schematically depicts an example of a system for isolating and reusing two or more reactant liquids and buffer solutions passed through a fluid device.

Fig. 2 schematically depicts a system performing a first reactant operation.

Fig. 3 schematically depicts a system that reverses the first reactant.

Fig. 4 schematically shows the system performing the first step of the first flushing operation.

Fig. 5 schematically shows the system performing the second step of the first flushing operation.

Fig. 6 schematically shows the system performing the reverse of the first buffer.

Fig. 7 schematically depicts a system performing a second reagent operation.

Fig. 8 schematically depicts a system that reverses the second reactant.

Fig. 9 schematically depicts a system performing the first step of a second flushing operation.

Fig. 10 schematically depicts a system performing the second step of a second flushing operation.

Fig. 11 schematically shows a system that reverses the second buffer.

Fig. 12 schematically depicts a system performing a third reagent operation.

Fig. 13 schematically depicts a system that reverses a third reactant.

Fig. 14 schematically shows the system performing the first step of the third flushing operation.

Fig. 15 schematically shows the system performing the second step of the third flushing operation.

Fig. 16 schematically shows a system that reverses the third buffer.

Fig. 17 schematically depicts a system performing a fourth reagent operation.

Fig. 18 schematically depicts a system that reverses a fourth reactant.

Fig. 19 schematically shows a system performing a fourth flushing operation.

Fig. 20 is a flow diagram of an exemplary method for isolating and reusing two or more reactant liquids and buffer solutions passed through a fluid device.

Fig. 21 is a block diagram of a system containing a liquid cartridge built into a processing instrument.

Implementation of the invention

Although the subject matter of the present invention may be embodied in various forms, the following description and accompanying drawings serve only to disclose some of these forms as specific examples. Accordingly, the subject matter of the invention is not intended to be limited to the forms or examples described and illustrated.

Unless otherwise stated, all technical terms, symbols and other technical terms or terminology used in the present description have the same meaning as generally accepted among specialists in the field to which the present invention relates.

Unless otherwise specified, or otherwise follows from the context, the element mentioned in the singular means "at least one" or "one or more".

In the present description, terms describing the relative spatial position and/or orientation can be used to characterize the position and/or orientation of a component, apparatus, location, design feature or part. Unless specifically stated or otherwise given by the context of the description, then, among other possibilities, such terms as top, bottom, above, below, under, above, top, bottom, left, right, front, behind, next, next to, between, horizontal, vertical, diagonal, longitudinal, transverse, radial, axial, and the like, are used for convenience in referring to such component, apparatus, location, feature or part in the drawings, and are not intended to limit the inventive concept.

In addition, unless otherwise noted, any specific dimensions mentioned in the description are given simply to provide an idea of an exemplary implementation of the device, i. implementation of aspects of the invention, and are not intended to limit the idea of the invention.

The term "approximately" applies to all numerical values given in the description, regardless of whether it is indicated explicitly or not. This term generally refers to a certain numerical range that a person skilled in the art would consider a reasonable amount of deviation from the mentioned numerical value (ie, a value that gives an equivalent result) in the context of the present invention. By way of example, which is not exhaustive, the term may be understood to include a deviation of ±10% from a given numerical value, provided that such deviation does not change the end result (or function) of that value. Therefore, under some circumstances, the specialist should be clear that approximately 1% can be understood as the range from 0.9% to 1.1%.

In the sense in which the term "adjacent" is used in the present description, it means "close" or "adjacent". Neighboring objects may be spaced apart from each other or may be in actual direct contact with each other. In some cases, neighboring objects may be connected to each other or may be integral with each other.

In the sense in which the terms "substantially" and "essential" are used in the present description, they characterize a significant measure or degree. When these terms are used, for example, in conjunction with an occurrence, circumstance, characteristic, or property, they may refer to situations in which the occurrence, circumstance, characteristic, or property occurs "exactly", as well as to situations in which the occurrence, circumstance, the characteristic or property takes place “with a reasonably good approximation”, for example, taking into account the tolerance or variability of the examples given in the description.

In the sense in which the terms "optional" and "optional" are used in the present description, they mean that the component, structure, element, event, circumstance, characteristic, property, etc. mentioned after them can be included in, and may or may not be included, or may or may not occur, and that the description covers situations in which a component, construct, element, event, circumstance, characteristic, property, etc. is included or occurs , and situations in which they are not included or do not occur.

In the sense in which the term "reagent" is used in the present description, it refers to any substance or combination of substances that are involved in molecular analysis, but other than sample material and analysis products. Examples of reagents include nucleotides, enzymes, amplification oligomers, probes, and salts.

As used herein, the term “buffer” refers to any pH controlled solution that can serve as a solvent for a solid (e.g., lyophilized) substance (e.g., reagent, sample, or combination) or as a diluent for diluting a liquid (eg, a reagent, a liquid sample, or a combination thereof; or a reagent solution, a sample, or a combination thereof).

In accordance with various examples, assemblies and devices, as discussed in the description, can be used in combination with a fluid cartridge, which may contain one or more fluid processing passages, including one or more elements, for example, one or more channels, a side channel, a valve, a flow splitter, a vent channel, a port, an access area, a transition, a drop, a drop containing a reagent, a coating layer, a reaction component, a combination of these elements, etc. . Any element can have fluid communication with another element.

All possible combinations of elements and components discussed in the description or mentioned in the claims are considered as part of the disclosure of the subject matter of the invention. It should be understood that all combinations of the above ideas and additional ideas, which will be discussed in more detail below, (provided that such ideas do not contradict each other), are considered as part of the subject matter of the invention disclosed in the present description. In particular, all combinations of claimed subject matter that appear at the end of this description are considered part of the disclosed subject matter.

In the appended claims, the term "comprising" is used as equivalent to the corresponding term "comprising". The terms "comprising" and "including" mean an open form that covers not only the listed elements, but also any additional elements. In addition, in the claims, the terms "first", "second", "third", etc. are used merely as an explanatory designation, and are not intended to impose any numerical requirements on the objects.

The term "fluid communication" means either direct fluid communication, for example, the two regions may be in fluid communication with each other through a fluid handling passage connecting the two said regions, or may be capable of fluid communication, for example, the two regions may be capable of fluid communication with each other when they are connected through a fluid handling passage, which may contain a valve located therein, and fluid communication can be established between these two areas when the valve is actuated, for example, by dissolving a soluble shutter, breaking a breakable damper, or otherwise opening a valve located in the fluid handling passage.

Fluid system

Although reusing buffer after flushing the fluid device reduces the amount of buffer needed for the sequencing process, the used buffer usually contains residual active reagent because sequencing procedures can instruct the fluid handling device to supply buffer solutions to the fluid device immediately after passing the active reagent through the fluid device. reagent. As a consequence, mixing of reused buffer with other fluid steps in the sequencing process can further contaminate the buffer supplied to the fluid device and/or impair the cleanliness of the fluid handling by inadvertently introducing residual reagents from the previous fluid handling with the recycled buffer. .

Thus, there is a need for improved fluid systems and methods that both isolate the buffer solution and reuse the buffer solution during the sequencing process to avoid contamination and reduce the volume of buffer solution that must be contained in the liquid cartridge for the sequencing process.

In accordance with various embodiments of the invention, the system is configured to isolate and reuse liquid reagents and buffer solutions passed through the fluid device, by providing one common buffer solution reservoir for storing fresh unused buffer solution, and at least one a specialized buffer tank designed to store the buffer solution that has already been used to remove each individual reagent when flushing the fluid device. The system may further comprise a flow control valve operatively associated with the common buffer reservoir and at least one dedicated buffer reservoir to selectively create a fluid connection of the fluid device with the common buffer reservoir or the dedicated buffer reservoir during the flushing operation. Accordingly, by isolating and reusing buffer through the use of one or more dedicated reservoirs, the system reduces the volume of buffer used in a fluid process such as sequencing, such as sequencing-by-synthesis (SBS) , which includes a splitting process, a cleaning process, an incorporation process, and a scanning process that are separated by one or more washing operations.

Fig. 1-19 depict an example of a system 100 for isolating and reusing a liquid reagent and buffer solution during two or more liquid operations. According to some examples, the system includes a fluid device 102, an inlet 103, a flow control valve 104, an outlet 105, a pump 106, a set of fluid reservoirs 110, and a set of 130 connecting channels. In some examples, system 100 is part of a fluid cartridge (not shown) that carries various components of system 100, such as fluid device 102, flow control valve 104, pump 106, and fluid reservoir stack 110, although one or more components of system 100 may or may not be mounted on a common fluid cartridge or other support structure.

As shown in FIG. 1, a fluid device 102 (e.g., a flow cell) is in fluid communication with a flow control valve 104 via an inlet 103. According to one example, the fluid device 102 is a flow cell comprising a first layer of glass (not shown) and a second layer of glass (not shown) that are bonded together to form one or more channels (not shown) between them through which liquids can be passed and optical detection can be performed. According to various examples, the fluid device 102 may include a fluid inlet 107, a fluid outlet 108, wherein one or more fluid channels (not shown) communicate with the fluid inlet and fluid outlet and allow fluid manipulation such as chemical or biochemical analysis, or carry out other processes or reactions. According to various examples, fluid device 102 is configured to allow fluid inlet 107 to be supplied with various types of fluids (eg, reagents, buffers, reaction media) for handling in one or more fluid channels. According to various examples, fluid device 102 is configured to further allow various types of fluids to be flushed out of one or more fluid channels through fluid outlet 108.

In the examples shown in FIG. 1-19, inlet 103 connects fluid inlet 107 of fluid device 102 to flow control valve 104, and outlet 105 connects fluid outlet 108 of fluid device 102 to pump 106. In other examples (not shown), system 100 may include two or more channels for connecting the fluid device 102 to the flow control valve 104, and two or more channels for connecting the fluid device 102 to the pump 106.

According to some examples, the liquid reservoir set 110 comprises two or more reservoirs 112, 114, 116, and/or 118 for liquid reactants. In the example shown in FIG. 1, these two or more liquid reactant tanks include: first liquid reactant tank 112, second liquid reactant tank 114, third liquid reactant tank 116, and fourth liquid reactant tank 118, although the present invention contemplates any number of liquid reactant tanks. The various reservoirs 112, 114, 116 and/or 118 for liquid reagents belonging to the set of reservoirs 110 may have the same or different sizes (i.e. volumes) - for example, all reservoirs 112, 114, 116 for liquid reagents and/or 118 may have the same volume, the liquid reactant tanks 112, 114, 116, and/or 118 may all have different volumes, or some subset of the liquid reactant tanks 112, 114, 116, and/or 118 may have the same volumes, and some subset of reactant tanks 112, 114, 116, and/or 118 may have different volumes depending on the required volume of reactant to be stored in each liquid reactant tank.

According to some examples, the reservoir 112 of the first liquid reagent contains the first liquid reagent 122 containing the solvent and the first reagent, intended for the first operation with the reagent (eg, splitting). In some examples, the second liquid reagent reservoir 114 contains a second liquid reagent 124 containing a solvent and a second reagent for a second reagent operation (eg, scanning). According to some examples, the reservoir 116 of the third liquid reagent contains a third liquid reagent 126 containing a solvent and a third reagent intended for the third operation with the reagent (eg, incorporation). In some examples, the fourth liquid reactant reservoir 118 contains a fourth liquid reactant 128 containing a solvent and a fourth reactant for a fourth reactant operation (eg, cleaning).

In some examples, fluid reservoir set 110 includes two or more buffer reservoirs 111, 113, 115, and/or 117. In the example shown in FIG. 1, said two or more buffer tanks include: a common buffer tank 111 and at least one specialized buffer tank 113, 115 and/or 117, each corresponding to one of the liquid reagents 122, 124, 126 and/ or 128. In the examples shown in FIG. 1-19, said at least one dedicated buffer reservoir is a first buffer reservoir 113, a second buffer reservoir 115, or a third buffer reservoir 117, although the invention contemplates any number of dedicated buffer reservoirs. The various buffer reservoirs 111, 113, 115 and/or 117 belonging to the fluid reservoir set 110 may have the same or different sizes (i.e. volumes)—for example, all buffer reservoirs 111, 113, 115 and/or 117 may have the same volume, the buffer reservoirs 111, 113, 115, and/or 117 may all have different volumes, or some subset of reservoirs 111, 113, 115, and/or 117 may have the same volumes, and the reservoirs a subset of reservoirs 111, 113, 115 and/or 117 may have different volumes depending on the required volume of liquid reactant to be stored in each liquid reactant reservoir.

In some examples, each of the dedicated buffer reservoirs 113, 115, and/or 117 contains a volume of fluid that is at least 30% of the volume of fluid that fluid device 102 contains. In some examples, each of the dedicated reservoirs 113, 115 and/or 117 for the buffer solution is a cache channel that has a constant cross-sectional dimension over its length. In some examples, each of the dedicated buffer reservoirs 113, 115, and/or 117 is a cache well that has a cross-sectional size larger than the cross-sectional size of its associated connection channel 130. In some examples, the cache well does not contain no sharp edges or features, and designed to minimize the formation of bubbles so that the cache well does not accumulate bubbles as fluid is pushed into and out of the reservoir.

In the examples shown in FIG. 1-19, the common buffer tank 111 contains a common buffer solution 121 (for example, a wash solution that contains salt water and soap) that was not used to flush the fluid device 102. According to various examples, each of the specialized tanks 113, 115 and/ or 117 for a buffer solution contains an isolated buffer solution that has already been used to flush the fluid device 102 after the fluid device 102 has been operated on with a reagent, for example, with one of the liquid reagents 122, 124 and/or 126, although initially, before any or reagent operations, dedicated buffer tanks 113, 115 and/or 117 may contain unused buffer.

In the examples shown in FIG. 6, the first buffer tank 113 contains the first buffer solution 123, which is a mixture of used buffer solution 602 that was passed through the fluid device 102 after the first liquid reagent 122 was directed into the fluid device 102 during the first reagent operation. In the examples shown in FIG. 11, the second buffer reservoir 115 contains a second buffer solution 125, which is a mixture of used buffer solution 1102 that was passed through the fluid device 102 after the second liquid reagent 124 was directed into the fluid device 102 during the second reagent operation. In the examples shown in FIG. 16, the third buffer reservoir 117 contains the third buffer solution 127, which is a mixture of used buffer solution 1602 that was passed through the fluid device 102 after the third liquid reagent 126 was directed into the fluid device 102 during the third reagent operation.

In some examples, the system may not include a dedicated buffer reservoir for each liquid reactant. For example, the system and processes depicted in FIG. 1-20 do not include a dedicated buffer reservoir corresponding to the fourth reactant liquid 128, while, in other examples, systems and processes may include a dedicated buffer reservoir corresponding to each reactant liquid.

In the examples shown in FIG. 1-19, the set of connection channels 130 comprises a respective connection 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 has a fluid connection with each liquid reservoir 111, 112, 113, 114, 115, 116, 117, or 118 included in the set 110 of liquid reservoirs. According to other examples (not shown), the set of 130 connecting channels may include two or more channels for fluid connection of the respective fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118 with a flow control valve 104, for example, a front a connection channel (such as the corresponding channel 131 depicted), and a rear connection channel (not shown) so that the used buffer solution can be returned for use to the rear side of the respective fluid reservoir 111, 112, 113, 114, 115, 116, 117 or 118, opposite the front connecting channel.

The flow control valve 104 is conceived and constructed to selectively fluidly connect one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 belonging to the fluid reservoir set 110 to the inlet 103, and thus to the fluid device 102. According to various examples, the flow control valve 104 is a rotary valve for selectively connecting to one of the connecting channels 131 of the respective liquid reservoir 111, 112, 113, 114, 115, 116, 117, or 118.

In the example shown in FIG. 1-19, the flow control valve 104 is a rotary valve comprising a rotary member 150 and a selector port 152. In some examples, the rotary member 150 is rotatable between a plurality of angle positions such that the valve selector port 152 can fluidly connect any one of fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 with a valve outlet port through the appropriate inlet port for each fluid reservoir. When the valve pivot 150 is rotated to a certain angular position so that the valve selector port 152 is aligned with one of the inlet ports for the selected fluid reservoir 111, 112, 113, 114, 115, 116, 117, or 118, fluid can move from the selected reservoir 111, 112, 113, 114, 115, 116, 117, or 118 through the valve selector port 152 and enter the valve outlet port.

In other examples (not shown), flow control valve 104 may be any other type of valve for selectively fluid connection of one of reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 to inlet 103. According to other examples ( not shown), the flow control valve 104 may be a series of valves, such as a plurality of pinch or solenoid valves with a manifold, to selectively connect one of the fluid reservoirs 111, 112, 113, 114, 115, 116, 117, or 118 to the inlet 103 .

In various examples, the pump 106, which is in fluid communication with the outlet port 105, is configured to create a pressure difference between any one reservoir 111, 112, 113, 114, 115, 116, 117, or 118 of the first set of fluid reservoirs 110 and the outlet 105 to allow bi-directional fluid flow along the respective connection channel 131 of the set of connection channels 130, through the flow control valve 104, through the inlet channel 103, the fluid device 102 and the outlet channel 105. The valve 106 may be a powered syringe pump (not shown), which is operatively associated with the syringe. According to various examples, the actuator is configured to move the syringe plunger in a first direction to create a negative pressure differential to draw liquid through the fluid device 102 towards (and possibly inward) the syringe barrel. The actuator is further configured to move the syringe plunger in a second direction, which is opposite to the first direction, to create a positive pressure difference to push fluid away from the syringe (and possibly from the syringe) towards the selected reservoir 111, 112, 113, 114, 115, 116, 117 or 118 from the first set of 110 liquid reservoirs. Accordingly, by moving the plunger in the second direction to create a positive pressure difference, the pump 106 can push the liquid contained in the fluid device 102 or the outlet port 105 through the inlet port 103, the flow control valve 104, the selected connection port 131, and supply said liquid. into one of the selected tanks 111, 112, 113, 114, 115, 116, 117 or 118.

In other examples (not shown), pump 106 may include any other differential pressure mechanism that is capable of reversing the direction of fluid flow.

The sequence of operations with fluids in the system

According to various examples, as shown in FIG. 2-19, system 100 isolates and reuses reagent and buffer solutions by: (i) selectively supplying a liquid reagent from any one of the liquid reagent tanks 112, 114, 116, and/or 118 to the fluid device 102 to perform the operation with reagent; (ii) optionally redirecting at least a portion of the used liquid reagent solution, i. e. 302, 802, 1302, or 1802 passed through fluid device 102 into a selected fluid reagent reservoir 112, 114, 116, and/or 118 for reuse in a subsequent reagent operation; (iii) selectively supplying a buffer solution from any buffer solution reservoir 111, 113, 115 and/or 117 to the fluid device 102 to perform a flushing operation; and (iv) redirecting at least a portion of the used buffer solution 602, 1102 and/or 1602 passed through the fluid device 102 back to one of the dedicated tanks 113, 115 or 117 for reuse in a subsequent flush operation.

According to FIG. 2, system 100 can be configured to perform a first reactant operation wherein flow control valve 104 allows fluid to flow from a selected first reactant liquid reservoir 112 belonging to reservoir set 110 into fluid device 102. During first reactant operation, control valve 104 flows is set in position to connect the reservoir 112 of the first liquid reagent with the fluid device 102 (for example, by connecting the selector channel 152 with a corresponding connection channel 131 associated with the reservoir 112 of the first liquid reagent). The pump 106 is actuated to draw the first liquid reactant 122 from the first liquid reactant reservoir 112 through the flow control valve 104 and the fluid device 102. As shown in FIG. 2, an aliquot 202 of the first liquid reactant 122 is passed through the appropriate connection channel 131, flow control valve 104, inlet 103, fluid device 102, outlet 105, and/or pump chamber 106.

According to FIG. 3, system 100 may be configured to perform a first liquid reactant reversal operation, wherein pump 106 redirects at least a portion of an aliquot 202 of first liquid reactant 122 that was transferred during the first reactant operation back to first liquid reactant reservoir 112. During the reversal of the first liquid reactant, the flow control valve 104 remains in position to connect the fluid device 102 to the first liquid reactant reservoir 112, and the pump 106 is actuated to push the liquid back through the fluid device 102 back into the first liquid reactant reservoir 112 . As shown in FIG. 3, at least a portion 302 of the aliquot 202 of the first liquid reactant 122 that was transferred in the first reactant operation is received back into the first liquid reactant reservoir 112 for reuse in one or more subsequent first reactant operations.

According to FIG. 4 and 5, the system 100 can be configured to perform a first flush operation wherein the flow control valve 104 initially allows fluid to flow from a selected first buffer reservoir 113 belonging to the reservoir set 110 into the fluid device 102 and then optionally allows fluid pass from the common buffer tank 111 belonging to the set of tanks 110 into the fluid device 102.

As shown in FIG. 4, during the first part of the first flush operation, the flow control valve 104 is set to connect the first buffer tank 113 to the fluid device 102 (for example, by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first buffer tank 113) . The pump 106 is actuated to suck an aliquot of the first buffer solution 123 from the first buffer solution reservoir 113 through the fluid device 102. As shown in FIG. 4, the first volume 402 of the first buffer solution 123 is transferred through the respective connecting channel 131, the flow control valve 104, the inlet 103, the fluid device 102, the outlet channel 105 and/or into the pump chamber 106 to flush out the first liquid reactant 122 remaining in the fluid device. 102. According to some embodiments of the invention, the first volume 402 may include the volume 602 of the reused first buffer solution shown in FIG. 6, in cases where the first buffer solution 123 has previously been pumped into the respective connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105 and/or pump chamber 106.

As shown in FIG. 5, according to some examples, during the second part of the first flush operation after the fluid device 102 has been flushed with the first volume 402 of the first buffer solution 123, the flow control valve 104 is set to connect the common buffer reservoir 111 to the fluid device 102 (for example , by connecting the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the tank 111 of the common buffer solution). The pump 106 is operated to draw an aliquot of the common buffer solution 121 from the common buffer reservoir 111 through the fluid device 102. As shown in FIG. 5, the second volume 502 of the common buffer solution 121 may be conveyed through an appropriate connection channel 131, a flow control valve 104, an inlet 103, a fluid device 102, an outlet 105, and/or a pump chamber 106. In some embodiments, the second volume 502 may mix with the first volume 402 in one or more of the following: an appropriate connection channel 131, a flow control valve 104, an inlet 103, a fluid device 102, an outlet 105, or a pump chamber 106. Thus, if the first volume 402 includes the volume 602 of the reused first buffer solution shown in FIG. 6, the inclusion of a common buffer solution 121 may dilute or otherwise reduce the presence of residues of the first liquid reagent 122 in the reused first buffer solution. Additionally, by supplying the common buffer solution 121 after the recycled first buffer solution, the common buffer solution 121 can in the fluid stream drive the recycled first buffer solution downstream of the fluid device 102.

According to other examples, the second part of the first flush operation illustrated in FIG. 5 may be omitted, and for the first flush operation, only the first buffer solution 123 may be sucked from the first buffer reservoir 113.

According to FIG. 6, the system 100 can be configured to perform a first buffer solution reversal operation in which the flow control valve 104 allows the liquid to move towards the first buffer reservoir 113. The pump 106 redirects the third volume 602 of the used buffer solution containing the first buffer solution 123 and/or the total buffer solution 121 back to the reservoir 113 of the first buffer solution. The third volume 602 may include at least a portion of the remainder of the liquid reagent 122 in addition to the first buffer solution 123 and/or the total buffer solution 121. In some cases, the percentage of the total buffer solution 121 in the third volume 602 is greater than the content of the first buffer solution 123 and/ or reused first buffer solution. During the reversal operation of the first buffer solution, the flow control valve 104 is set to connect the fluid device 102 to the first buffer tank 113 (for example, by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first buffer tank 113), and the pump 106 is actuated to move the liquid in the reverse direction through the fluid device 102 to the reservoir 113 of the first buffer solution. As shown in FIG. 6, the third volume 602 of used buffer is received back into the first buffer reservoir 113 for reuse in one or more subsequent first flush operations. According to some examples, the third volume 602 of the used buffer solution is less than the second volume 502 or equal to the second volume 502 transferred from the common buffer tank 111, and according to other examples, the third volume 602 of the used buffer solution is greater than the second volume 502 transferred from the common buffer tank 111 solution, and according to other examples, the third volume 602 of the used buffer solution is equal to the total amount of buffer solution passed through the fluid device 102, i.e. taken together the first volume 202 transferred from the first buffer tank 113 and the second volume 502 transferred from the common buffer tank 111.

According to FIG. 7, system 100 can be configured to perform a second reactant operation wherein flow control valve 104 allows fluid to flow from a selected second reactant liquid reservoir 114 belonging to reservoir set 110 into fluid device 102. During second reactant operation, control valve 104 flows is set in position to connect the tank 114 of the second liquid reagent with the fluid device 102 (for example, by connecting the selector channel 152 with a corresponding connection channel 131 associated with the tank 114 of the second liquid reagent). The pump 106 is actuated to draw the second liquid reactant 124 from the second liquid reactant reservoir 114 through the flow control valve 104 and the fluid device 102. As shown in FIG. 7, an aliquot 702 of the first liquid reactant 124 is passed through the appropriate connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105, and/or into the pump chamber 106.

According to FIG. 8, system 100 can be configured to perform a second reagent reversal operation, wherein pump 106 redirects at least a portion of an aliquot 702 of second liquid reagent 124 that was transferred during the second reagent operation back to second liquid reagent reservoir 114. During the reversal of the second liquid reactant, the flow control valve 104 remains in position to connect the fluid device 102 to the second liquid reactant reservoir 114, and the pump 106 is actuated to push the liquid back through the fluid device 102 back into the second liquid reactant reservoir 114 . As shown in FIG. 8, at least a portion 802 of the aliquot 702 of the second liquid reagent 124 that was transferred in the second reagent operation is received back into the second liquid reagent reservoir 114 for reuse in one or more subsequent second reagent operations.

According to FIG. 9 and 10, the system 100 may be configured to perform a second flush operation wherein the flow control valve 104 initially allows fluid to flow from a selected second buffer reservoir 115 belonging to the reservoir set 110 into the fluid device 102 and then optionally allows fluid pass from the common buffer tank 111 belonging to the set of tanks 110 into the fluid device 102.

As shown in FIG. 9, during the first part of the second flush operation, the flow control valve 104 is set in position to connect the second buffer tank 115 to the fluid device 102 (for example, by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the second buffer tank 115) . Pump 106 is actuated to draw an aliquot of second buffer 124 from second buffer reservoir 115 through fluid device 102. As shown in FIG. 9, the fourth volume 902 of the second buffer solution 124 is transferred through the respective connecting channel 131, the flow control valve 104, the inlet channel 103, the fluid device 102, the outlet channel 105 and/or the pump chamber 106 to flush out the second liquid reagent 124 remaining in the fluid device. 102. In some embodiments, the fourth volume 902 may include a reused second buffer volume 1102 as shown in FIG. 11, in cases where the second buffer solution 125 has previously been pumped into the respective connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105 and/or pump chamber 106.

As shown in FIG. 10, according to some examples, during the second part of the second flush operation after the fluid device 102 has been flushed with the fourth volume 902 of the second buffer solution 125, the flow control valve 104 is set to connect the common buffer reservoir 111 to the fluid device 102 (for example , by connecting the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the tank 111 of the common buffer solution). The pump 106 is operated to draw an aliquot of the common buffer solution 121 from the common buffer reservoir 111 through the fluid device 102. As shown in FIG. 10, the fifth volume 1002 of the total buffer solution 121 may be transferred through the appropriate connection channel 131, the flow control valve 104, the inlet 103, the fluid device 102, the outlet 105 and/or the pump chamber 106. In some embodiments, the fifth volume 1002 may mix with the fourth volume 902 in one or more of the following: an appropriate connection channel 131, a flow control valve 104, an inlet 103, a fluid device 102, an outlet 105, or a pump chamber 106. Thus, if the fourth volume 902 includes volume 1102 of the reused second buffer solution shown in FIG. 11, the inclusion of a common buffer solution 121 may dilute or otherwise reduce the presence of residues of the second liquid reagent 124 in the recycled second buffer solution. Additionally, by supplying the common buffer solution 121 after the recycled second buffer solution, the common buffer solution 121 can in the fluid stream drive the recycled second buffer solution downstream of the fluid device 102.

According to other examples, the second part of the second flush operation illustrated in FIG. 10 may be omitted, and for the second flush operation, only the second buffer solution 125 may be drawn from the second buffer reservoir 115.

According to FIG. 11, system 100 can be configured to perform a second buffer reversal operation in which flow control valve 104 allows fluid to move towards second buffer reservoir 115. The pump 106 redirects the sixth volume 1102 of the used buffer solution containing the second buffer solution 125 and/or the total buffer solution 121 back to the tank 115 of the second buffer solution. The sixth volume 1102 may include at least a portion of the remainder of the second liquid reagent 124 in addition to the second buffer solution 125 and/or the total buffer solution 121. In some cases, the percentage of the total buffer solution 121 in the sixth volume 1102 is greater than the content of the second buffer solution 125 and /or reused second buffer solution. During the reversal operation of the second buffer solution, the flow control valve 104 is set to connect the fluid device 102 to the second buffer reservoir 115 (for example, by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the second buffer reservoir 115), and the pump 106 is actuated to move the liquid in the reverse direction through the fluid device 102 to the tank 115 of the second buffer solution. As shown in FIG. 11, a sixth volume 1102 of used buffer is received back into second buffer reservoir 115 for reuse in one or more subsequent second flush operations. According to some examples, the sixth volume 1102 of the used buffer solution is less than the fifth volume 1002 or equal to the fifth volume 1002 transferred from the common buffer tank 111, and according to other examples, the sixth volume 1102 of the used buffer solution is greater than the fifth volume 1002 transferred from the common buffer tank 111 solution, and according to other examples, the sixth volume 1102 of the used buffer solution is equal to the total amount of buffer solution passed through the fluid device 102, i.e. taken together, the fourth volume 902 transferred from the second buffer tank 115 and the fifth volume 1002 transferred from the common buffer tank 111.

According to FIG. 12, system 100 may be configured to perform a third reactant operation wherein flow control valve 104 allows fluid to flow from a selected third reactant liquid reservoir 116 belonging to reservoir set 110 into fluid device 102. During third reactant operation, control valve 104 flows is set in position to connect the reservoir 116 of the third liquid reagent with the fluid device 102 (for example, by connecting the selector channel 152 with a corresponding connection channel 131 associated with the reservoir 116 of the third liquid reagent). Pump 106 is actuated to draw third liquid reactant 126 from third liquid reactant reservoir 116 through flow control valve 104 and fluid device 102. As shown in FIG. 12, an aliquot 1202 of the third liquid reactant 126 is passed through the appropriate connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105, and/or into the pump chamber 106.

According to FIG. 13, system 100 can be configured to perform a third reagent reverse operation, wherein pump 106 redirects at least a portion of an aliquot 1202 of third liquid reagent 126 that was transferred during the third reagent operation back to third liquid reagent reservoir 116. During the reversal of the third liquid reactant, the flow control valve 104 remains in position to connect the fluid device 102 to the third liquid reactant reservoir 116, and the pump 106 is actuated to push the liquid back through the fluid device 102 back into the third liquid reactant reservoir 116 . As shown in FIG. 3, at least a portion 1302 of an aliquot 1202 of the third liquid reagent 126 that was transferred in the third reagent operation is received back into the third liquid reagent reservoir 116 for reuse in one or more subsequent third reagent operations.

According to FIG. 14 and 15, the system 100 may be configured to perform a third flush operation in which the flow control valve 104 initially allows fluid to flow from a selected third buffer reservoir 117 belonging to the reservoir set 110 into the fluid device 102 and then optionally allows fluid pass from the common buffer tank 111 belonging to the set of tanks 110 into the fluid device 102.

As shown in FIG. 14, during the first part of the third flush operation, the flow control valve 104 is set to connect the third buffer reservoir 117 to the fluid device 102 (for example, by connecting the valve selector port 152 to the corresponding connection port 131 associated with the third buffer reservoir 117) . Pump 106 is actuated to draw an aliquot of third buffer 127 from third buffer reservoir 117 through fluid device 102. As shown in FIG. 14, the seventh volume 1402 of the third buffer solution 127 is transferred through the respective connecting channel 131, the flow control valve 104, the inlet 103, the fluid device 102, the outlet channel 105 and/or the pump chamber 106 to flush out the third liquid reagent 126 remaining in the fluid device. 102. In some embodiments, the seventh volume 1402 may include the reused third buffer volume 1602 shown in FIG. 16, in cases where the third buffer solution 127 has previously been pumped into the respective connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105 and/or pump chamber 106.

As shown in FIG. 15, according to some examples, during the second part of the third flush operation after the fluid device 102 has been flushed with the seventh volume 1402 of the third buffer solution 127, the flow control valve 104 is set to connect the common buffer reservoir 111 to the fluid device 102 (for example , by connecting the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the tank 111 of the common buffer solution). The pump 106 is operated to draw an aliquot of the common buffer solution 121 from the common buffer reservoir 111 through the fluid device 102. As shown in FIG. 15, an eighth volume 1502 of the total buffer solution 121 may be transferred through the appropriate connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105, and/or pump chamber 106. In some embodiments, the eighth volume 1502 may mix with the seventh volume 1402 in one or more of the following: an appropriate connecting passage 131, a flow control valve 104, an inlet 103, a fluid device 102, an outlet 105, or a pump chamber 106. Thus, if the seventh volume 1402 includes volume 1602 of the reused third buffer solution shown in FIG. 16, the inclusion of common buffer solution 121 may dilute or otherwise reduce the presence of residual third liquid reagent 126 in the reused third buffer solution. Additionally, by supplying the common buffer solution 121 after the recycled third buffer solution, the common buffer solution 121 can in the fluid stream drive the recycled third buffer solution downstream of the fluid device 102.

According to other examples, the second part of the third washing operation illustrated in FIG. 15 may be omitted, and for the third flush operation, only the third buffer solution 127 may be drawn from the third buffer reservoir 117.

According to FIG. 16, system 100 can be configured to perform a third buffer reverse operation in which flow control valve 104 allows fluid to flow towards third buffer reservoir 117. The pump 106 redirects the ninth volume 1602 of the used buffer solution containing the third buffer solution 127 and/or the total buffer solution 121 back to the reservoir 117 of the third buffer solution. The ninth volume 1602 may include at least a portion of the remainder of the third liquid reagent 126 in addition to the third buffer solution 127 and/or the total buffer solution 121. In some cases, the percentage of the total buffer solution 121 in the ninth volume 1602 is greater than the content of the third buffer solution 127 and /or reused third buffer solution. During the third buffer reversal operation, the flow control valve 104 is set to connect the fluid device 102 to the third buffer reservoir 117 (for example, by connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the third buffer reservoir 117), and the pump 106 is actuated to move the liquid in the reverse direction through the fluid device 102 to the reservoir 117 of the third buffer solution. As shown in FIG. 16, a ninth volume 1602 of used buffer is received back into third buffer reservoir 117 for reuse in one or more subsequent third flush operations. According to some examples, the ninth volume 1602 of the used buffer solution is less than the eighth volume 1502 or equal to the eighth volume 1502 transferred from the common buffer tank 111, and according to other examples, the ninth volume 1602 of the used buffer solution is greater than the eighth volume 1502 transferred from the common buffer tank 111 solution, and according to other examples, the ninth volume 1602 of the used buffer solution is equal to the total amount of buffer solution passed through the fluid device 102, i.e. taken together the seventh volume 1402 transferred from the third buffer tank 117 and the eighth volume 1502 transferred from the common buffer tank 111.

According to FIG. 17, the system 100 may be configured to perform a fourth reactant operation wherein the flow control valve 104 allows fluid to flow from a selected fourth reactant liquid reservoir 118 belonging to the reservoir set 110 into the fluid device 102. During fourth reactant operation, the control valve 104 flows is set in position to connect the reservoir 118 of the fourth liquid reagent with the fluid device 102 (for example, by connecting the selector channel 152 with a corresponding connection channel 131 associated with the reservoir 118 of the fourth liquid reagent). Pump 106 is actuated to draw fourth liquid reactant 128 from fourth liquid reactant reservoir 118 through flow control valve 104 and fluid device 102. As shown in FIG. 17, an aliquot 1702 of the fourth liquid reactant 128 is passed through the appropriate connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105, and/or pump chamber 106.

According to FIG. 18, system 100 can be configured to perform a fourth reactant liquid reversal operation in which pump 106 redirects at least a portion of an aliquot 1702 of fourth reactant liquid 128 that was transferred during the fourth reactant operation back to fourth reactant liquid reservoir 118. During the reversal of the fourth liquid reactant, the flow control valve 104 remains in position to connect the fluid device 102 to the fourth liquid reactant reservoir 118, and the pump 106 is actuated to push the liquid back through the fluid device 102 back into the fourth reactant liquid reservoir 118 . As shown in FIG. 18, at least a portion 1802 of the aliquot 1702 of the fourth liquid reagent 128 that was transferred in the fourth reagent operation is received back into the fourth liquid reagent reservoir 118 for reuse in one or more subsequent fourth reagent operations.

According to some examples, it may be unacceptable or impractical to reuse a buffer solution after a wash operation - for example, if the characteristics of the reagent or other material that is washed out during the wash operation are such that the risk of carryover associated with reusing the buffer that was used is too great. to wash out the reagent. In one example, combining the buffer used to flush reagents in the fluid device 102 after a SBS purification process with fresh buffer typically disrupts subsequent SBS processes, even if a trace amount of used buffer is added to the fresh buffer. In such a situation, buffer from the common buffer tank 111 can be passed through the fluid device, but the used buffer is not then returned back to the isolated buffer tank. In some examples, the decision that the risk of reusing a buffer solution is too great is made when the activity of the used buffer solution exceeds a first level of contamination that can be determined experimentally.

In some examples, system 100 may reuse buffer solution from the same reservoir in two different wash operations that follow two different reagent operations. For example, if the characteristics of a reagent or other material in a first reagent operation are non-hazardous, meaning that the presence of that reagent or material will not affect or interfere with another reagent operation (e.g., a second reagent operation), then the buffer solution used to flush out said reagent or material after the first reagent operation can be used again in the wash operations that follow the second reagent operation. In some examples, a judgment that a used buffer solution is not hazardous is made when the activity of the used buffer solution is below a second contamination level that can be determined experimentally.

According to FIG. 19, system 100 may be configured to perform a fourth flush operation in which flow control valve 104 allows fluid to flow from a selected common buffer reservoir 111 belonging to reservoir set 110 into fluid device 102. As shown in FIG. 19, the volume 1902 of the total buffer solution 121 is transferred through the respective connecting channel 131, the flow control valve 104, the inlet 103, the fluid device 102, the outlet channel 105 and/or the pump chamber 106 to flush out the fourth liquid reagent 128 remaining in the fluid device 102 However, the system 100 does not contain a reservoir to store and isolate the buffer used in the fourth wash operation for possible reuse. Thus, the volume 1902 of buffer passed through the fluid device during the fourth flush operation is not transferred back from the respective connection channel 131, flow control valve 104, inlet channel 103, fluid device 102, outlet channel 105, and/or pump chamber 106. In contrast, , this buffer solution can be pumped through the appropriate connection port 131, flow control valve 104, inlet port 103, fluid device 102, outlet port 105 and/or pump chamber 106 and discharged into a waste reservoir (not shown).

Method for isolating and reusing liquid reagent and buffer solution

According to various examples, FIG. 20 illustrates a method 2000 for isolating and reusing liquid reagents and buffer solutions.

The method 2000 includes the step of 2010 moving an aliquot of the first liquid reactant 122 into the fluid device 102. According to some examples, the step 2010 includes setting the flow control valve 104 to a position where the valve allows fluid to pass from the first liquid reactant reservoir 112 into the fluid device 102. In some examples, For example, step 2010 includes turning on a motor to rotate the valve 104 until aligned and fluidly connecting the valve selector channel 152 to a corresponding connection channel 131 associated with the first liquid reactant reservoir 112. In some examples, step 2010 includes using a pump 106 to move the first liquid reactant 122 from reservoir 112 to fluid device 102. In some examples, step 2010 includes using an actuator to move the plunger of syringe pump 106 in a first direction to create a negative differential pressure to move first liquid reactant 122 through fluid device 102.

Method 2000 may further comprise a first step 2015 of moving a volume of buffer solution into fluid device 102. According to some examples, step 2015 includes a first part comprising moving a volume of first buffer solution 123 from reservoir 113 to fluid device 102. According to some examples, the first part step 2015 comprises setting the flow control valve 104 to a position that allows fluid from the first buffer reservoir 113 to flow into the fluid device 102, and using a pump 106 to move the first buffer solution 123 from the reservoir 113 to the fluid device 102. According to some examples, the first part step 2015 comprises turning on a motor to rotate the flow control valve 104 until aligned and fluidly connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first buffer reservoir 113. In some examples, the first part of step 2015 comprises using an actuator to move the syringe pump plunger 106 in a first direction to create a negative pressure differential to move the first buffer solution 123 through the fluid device 102.

According to some examples, step 2015 includes a second part comprising (after moving the first buffer solution 123) moving a volume of common buffer solution 121 from reservoir 111 to fluid device 102. According to some examples, the second part of step 2015 includes placing a flow control valve 104 in a position that allows fluid from the common buffer reservoir 111 to flow into the fluid device 102. According to some examples, the second part of step 2015 comprises turning on a motor to rotate the flow control valve 104 until aligned and fluidly connecting the valve selector channel 152 to the corresponding connection channel 131 associated with a reservoir 111 of a common buffer solution. According to some examples, the second part of step 2015 comprises using an actuator to move the plunger of the syringe pump 106 in a first direction to create a negative pressure difference to move the common buffer solution 121 of the buffer solution 123 through the fluid device 102.

In some examples, the volume of first buffer 123 moved out of tank 113 is substantially equal to the volume of total buffer 121 moved out of tank 111. In other cases, the volume of first buffer 123 moved out of tank 113 is greater than the volume of total buffer 121 moved from tank 111. And in other cases, the volume of the first buffer solution 123 transferred from the tank 113 is less than the volume of the total buffer solution 121 transferred from the tank 111.

The method 2000 may further comprise the step of 2020 moving at least a portion of the buffer solution volume transferred in step 2015 to the first buffer tank 113. According to some examples, step 2020 comprises setting the flow control valve 104 to a position that allows fluid from the fluid device 102 to flow into the first buffer reservoir 113. According to some examples, step 2020 comprises turning on a motor to rotate the flow control valve 104 until aligned and fluidly connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the first buffer reservoir 113. According to some examples, step 2020 comprises using a pump 106 to push the buffer solution from the fluid device 102 into the reservoir 113 of the first buffer solution. In some examples, step 2020 comprises using an actuator to move the syringe pump plunger in a second direction to create a positive differential pressure to move the buffer into the first buffer reservoir 113. According to some examples, the proportion of buffer solution transferred to reservoir 113 at step 2020 is in the range of approximately 30% to 70% of the total volume of buffer solution transferred to fluid device 102 at step 2015. For example, if 50 μl is transferred from reservoir 113 of the first buffer solution 123, and 50 µl of the total buffer solution 121 is transferred from the tank 111, then the volume transferred back to the first buffer solution tank 113 can be from 30 µl to 70 µl out of a total volume of 100 µl. In some cases, the volume transferred back to the first buffer reservoir 113 may contain predominantly the total buffer solution 121, given that the total buffer solution displaces the first buffer solution 123 further downstream in the fluid system.

The method 2000 may further comprise the step of 2025 moving an aliquot of the second liquid reactant 124 into the fluid device 102. According to some examples, step 2025 comprises setting the flow control valve 104 to a position such that the valve allows liquid to flow from the second liquid reactant reservoir 114 into the fluid device 102. According to some examples, step 2025 comprises turning on a motor to rotate valve 104 until aligned and fluidly connecting valve selector channel 152 to a corresponding connection channel 131 associated with second liquid reactant reservoir 114. In some examples, step 2025 comprises using pump 106 to move second liquid reactant 124 from reservoir 114 to fluid device 102. In some examples, step 2025 includes using an actuator to move the plunger of syringe pump 106 in a first direction to create a negative pressure differential to move second liquid reactant 124 through fluid device 102.

Method 2000 may further comprise a second step 2030 of moving a volume of buffer solution into fluid device 102. According to some examples, step 2030 includes a first part comprising moving a volume of second buffer solution 125 from reservoir 115 to fluid device 102. In some examples, the first part step 2030 includes setting the flow control valve 104 to a position that allows fluid from the second buffer reservoir 115 to flow into the fluid device 102, and using a pump 106 to move the second buffer solution 125 from the reservoir 115 to the fluid device 102. According to some examples, the first part step 2030 includes turning on a motor to rotate the flow control valve 104 until aligned and fluidly connect the valve selector channel 152 to the corresponding connection channel 131 associated with the second buffer reservoir 115. In some examples, the first part of step 2030 comprises using an actuator to move the plunger of the syringe pump 106 in a first direction to create a negative pressure differential to move the second buffer solution 125 through the fluid device 102.

According to some examples, step 2030 includes a second part comprising (after moving the second buffer 125) moving a volume of common buffer 121 from reservoir 111 to fluid device 102. According to some examples, the second part of step 2030 includes placing a flow control valve 104 in a position that allows fluid from the common buffer reservoir 111 to flow into the fluid device 102; and using a pump 106 to draw a volume of the common buffer solution 121 from the reservoir 111 into the fluid device 102. According to some examples, the second part of step 2030 comprises turning on a motor to rotate the valve 104 flow control until alignment and fluid connection of the valve selector channel 152 with the corresponding connecting channel 131 associated with the reservoir 111 of the common buffer solution. In some examples, the second part of step 2030 comprises using an actuator to move the plunger of the syringe pump 106 in a first direction to create a negative pressure differential to move the common buffer solution 121 through the fluid device 102.

The method 2000 may further comprise a step 2035 of moving at least a portion of the volume of buffer solution transferred in step 2030 into the reservoir 115 of the second buffer solution. According to some examples, step 2035 comprises setting the flow control valve 104 to a position that allows fluid from the fluid device 102 to flow into the reservoir 115 of the second buffer solution. According to some examples, step 2035 comprises turning on a motor to rotate the flow control valve 104 until aligned and fluidly connecting the valve selector channel 152 to a corresponding connection channel 131 associated with the second buffer reservoir 115. According to some examples, step 2035 comprises using a pump 106 to push the buffer solution from the fluid device 102 into the reservoir 115 of the second buffer solution. In some examples, step 2035 comprises using an actuator to move the syringe pump plunger in a second direction to create a positive differential pressure to move the buffer into the second buffer reservoir 115. In some examples, the proportion of buffer transferred to reservoir 115 at step 2035 is in the range of approximately 30% to about 70% of the total volume of buffer transferred to fluid device 102 at step 2030. For example, if 50 μl is transferred from reservoir 115 of the second buffer solution 125, and 50 μl of the total buffer solution 121 is transferred from the reservoir 111, then the volume transferred back to the second buffer reservoir 115 can be from 30 μl to 70 μl out of a total volume of 100 μl. In some cases, the volume transferred back to the second buffer reservoir 115 may contain predominantly the common buffer 121, given that the common buffer displaces the second buffer 125 further downstream in the fluid system.

Further, the method 2000 may comprise the step 2040 of reusing the first liquid reactant 122 by moving an aliquot of the first liquid reactant 122 into the fluid device 102. According to some examples, the step 2040 comprises setting the flow control valve 104 to a position where the valve allows liquid to flow from the reservoir 112 of the first fluid reactant into fluid device 102. In some examples, step 2040 comprises turning on a motor to rotate valve 104 until aligned and fluidly connecting valve selector channel 152 to a corresponding connection channel 131 associated with first fluid reactant reservoir 112. In some examples, step 2040 includes using a pump 106 to move the first liquid reactant 122 from reservoir 112 to fluid device 102. In some examples, step 2040 includes using an actuator to move the plunger of syringe pump 106 in a first direction to create a negative pressure differential to move first liquid reactant 122 through fluid device 102.

The method 2000 may further comprise the step of reusing the first buffer solution 2045 by moving a volume of buffer solution into the fluid device 102, wherein at least a portion of the volume of buffer solution moved in step 2045 is transferred from the first buffer reservoir 113, which now contains the used buffer solution that has been transferred to the first buffer reservoir 113 in step 2020. According to some examples, step 2045 includes a first portion comprising setting the flow control valve 104 to a position where the valve allows fluid to flow from the first buffer reservoir 113 to fluid device 102. According to some examples, the first part of step 2045 comprises setting the flow control valve 104 to a position where the valve allows fluid to flow from the first buffer solution reservoir 113 into the fluid device 102, and using a pump 106 to move the first buffer solution 123 out of the reservoir 113 into the fluid device 102. According to some examples, the first part of step 2045 comprises turning on a motor to rotate the valve 104 until aligned and fluidly connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the reservoir 113 of the first buffer solution. In some examples, the first part of step 2045 comprises using an actuator to move the syringe pump plunger 106 in a first direction to create a negative pressure differential to move the first buffer solution 123 through the fluid device 102.

According to some examples, step 2045 includes a second part comprising (after moving the first buffer solution) moving a volume of common buffer solution 121 from reservoir 111 to fluid device 102. According to some examples, the second part of step 2045 includes placing a flow control valve 104 in a position where the valve allows fluid to pass from the common buffer reservoir 111 into the fluid device 102; and using a pump 106 to move the common buffer solution 121 from the reservoir 111 into the fluid device 102. According to some examples, the second part of step 2045 comprises turning on a motor to rotate valve 104 until alignment and liquid connection of the valve selector channel 152 with the corresponding connecting channel 131 associated with the reservoir 111 of the common buffer solution. In some examples, the second part of step 2045 comprises using an actuator to move the plunger of the syringe pump 106 in a first direction to create a negative pressure differential to move the common buffer solution 121 through the fluid device 102. In some examples, the volume of the first buffer solution 123 moved from the reservoir 113 is essentially equal to the volume of the total buffer solution 121 moved from the tank 111.

In some examples, if the first buffer solution 123 is to be reused after step 2045, then the method comprises the step of moving at least a portion of the volume of buffer solution transferred in step 2045 back to the first buffer reservoir 113, as in step 2020 discussed above. . According to some examples, the step of moving at least a portion of the volume of buffer solution transferred at step 2045 back to reservoir 113 comprises setting the flow control valve 104 to a position such that the valve allows fluid to flow from the fluid device 102 into the reservoir 113 of the first buffer solution. According to some examples, the step of moving at least a portion of the volume of the buffer solution transferred at step 2045 back to the tank 113 includes turning on the motor to rotate the valve 104 until the alignment and fluid connection of the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the tank 113 of the first buffer solution. According to some examples, the step of moving at least a portion of the volume of buffer solution transferred in step 2045 back to reservoir 113 comprises using an actuator to move the plunger of syringe pump 106 in a second direction to create a positive pressure difference to move the buffer solution into reservoir 113 of the first buffer solution. According to some examples, the proportion of the buffer solution transferred from the first buffer reservoir 113 at step 2045 is in the range of approximately 30% to 70% of the volume of buffer solution transferred to the fluid device at step 2045.

The method 2000 may further comprise the step 2050 of reusing the second liquid reactant 124 by moving an aliquot of the second liquid reactant 124 into the fluid device 102. According to some examples, the step 2050 comprises setting the flow control valve 104 to a position where the valve allows liquid to pass from the reservoir 114 of the second liquid reactant to fluid device 102. In some examples, step 2050 comprises turning on a motor to rotate valve 104 until aligned and fluidly connecting valve selector channel 152 to a corresponding connection channel 131 associated with second fluid reactant reservoir 114. In some examples, step 2050 comprises using a pump 106 to draw and move second liquid reactant 124 from reservoir 114 to fluid device 102. In some examples, step 2050 includes using an actuator to move a plunger of syringe pump 106 in a first direction to create a negative pressure difference to move the second liquid reactant 124 through the fluid device 102.

The method 2000 may further comprise the step of reusing the second buffer solution 2055 by moving a volume of buffer solution into the fluid device 102, wherein at least a portion of the volume of buffer solution moved in step 2055 is transferred from the second buffer reservoir 115, which now contains the used buffer solution that has been transferred to the second buffer reservoir 115 in step 2035. According to some examples, step 2055 includes a first portion comprising setting the flow control valve 104 to a position where the valve allows fluid to flow from the second buffer reservoir 115 to fluid device 102. According to some examples, the first part of step 2055 comprises setting the flow control valve 104 to a position where the valve allows fluid to flow from the second buffer solution reservoir 115 into the fluid device 102, and using a pump 106 to move the second buffer solution 125 out of the reservoir 115 into the fluid device 102. According to some examples, the first part of step 2055 comprises turning on a motor to rotate the valve 104 until aligned and fluidly connecting the valve selector channel 152 to the corresponding connection channel 131 associated with the second buffer reservoir 115. In some examples, the first part of step 2055 comprises using an actuator to move the plunger of the syringe pump 106 in a first direction to create a negative pressure differential to move the second buffer solution 125 through the fluid device 102.

According to some examples, step 2055 includes a second part comprising (after moving the second buffer solution 125) moving a volume of total buffer solution 121 from reservoir 111 to fluid device 102. According to some examples, the second part of step 2055 includes setting the flow control valve 104 to a position where the valve allows fluid to pass from the common buffer reservoir 111 to the fluid device 102, and using a pump 106 to move the common buffer solution 121 from the reservoir 111 to the fluid device 102. According to some examples, the second part of step 2055 comprises turning on a motor to rotating the valve 104 until aligned and fluidly connecting the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the tank 111 of the common buffer solution. In some examples, the second part of step 2055 comprises using an actuator to move the syringe pump plunger 106 in a first direction to create a negative pressure differential to move the common buffer solution 121 through the fluid device 102. In some examples, the volume of the second buffer solution 125 moved from the reservoir 115 is essentially equal to the volume of the total buffer solution 121 moved from the tank 111.

In some examples, if further reuse of the second buffer solution 125 after step 2055 is contemplated, then the method comprises the step of moving at least a portion of the volume of buffer solution transferred in step 2055 back to the second buffer reservoir 115, as in step 2035 discussed above. . According to some examples, the step of moving at least a portion of the volume of buffer solution transferred in step 2055 back to reservoir 115 comprises setting the flow control valve 104 to a position such that the valve allows fluid to flow from the fluid device 102 to the reservoir 115 of the second buffer solution. According to some examples, the step of moving at least a portion of the volume of the buffer solution transferred in step 2055 back to the reservoir 115 comprises turning on a motor to rotate the valve 104 until aligned and fluidly connecting the selector channel 152 of the valve with the corresponding connecting channel 131 associated with the reservoir 115 of the second buffer solution. According to some examples, the step of moving at least a portion of the volume of buffer solution transferred at step 2055 back into reservoir 115 comprises using an actuator to move the plunger of syringe pump 106 in a second direction to create a positive pressure difference to move the buffer solution into reservoir 115 of the second buffer solution.

processing device

As shown schematically in FIG. 21, in accordance with several embodiments of the invention, system 100 includes a fluid cartridge 52 that carries various components of system 100 such as a fluid device 102, a flow control valve 104, a pump 106, and a set of fluid reservoirs 110. Fluid cartridge 52 can be operatively installed in processing apparatus 50. In some examples, fluid cartridge 52 includes an inlet 103, a flow control valve 104, at least a portion of an outlet 105, a set of fluid reservoirs 110, and a set of connection channels 130. Fluid cartridge 52 may be operatively coupled to instrument 50, wherein instrument 50 may include one or more actuators (e.g., a motor) to control the position of flow control valve 104 and the pressure generated by pump 106 to select and move various reagent solutions (liquid reagents). ) and buffer solutions. Instrument 50 may further include a drain port (not shown) for draining spent liquid reagents and buffer solutions. Controller 140, which may be part of instrument 50, or may be a separate device or remote computer resource operatively connected to instrument 50, controls the operation of instrument 50 (i.e., operation of fluid device 102 and pump 106) as well as operation of the fluid cartridge. 52 (eg, operation of the flow control valve 104).

Hardware and software

The present invention in its aspects is implemented by control and computing hardware components, user generated software, data input components, and data output components. Hardware components include computing and control modules (e.g., system controller(s)), such as microprocessors and computers, configured to perform computing and/or control steps by receiving one or more input values, executing one or more algorithms, such as the algorithm shown in Fig. 20 stored on a non-volatile computer-readable medium (eg, programs) that provide instructions for manipulating or otherwise influencing input values, and outputting one or more output values. Such output values may be displayed or otherwise presented to the user as user-relevant information, such as information about the state of the instrument or the process being performed by the instrument, or such output quantities may be input to other processes and/or control algorithms. Input components contain elements through which data is entered for subsequent use by control and computing hardware components. Such inputs may include encoder signals, motor encoders, as well as manual input signal elements such as graphical user interfaces, keyboards, touch screens, microphones, switches, manual scanners, voice-triggered signals, and the like. Data output components may include hard drives or other storage media, graphical user interfaces, monitors, printers, indicator lights or audible alarms (eg, vibrator, siren, bell, etc.). The software contains instructions stored on a non-volatile computer-readable storage medium that, when executed by the control and computing hardware components, cause the control and computing hardware to perform one or more automatic or semi-automatic processes.

According to some examples, the instrument may include a control system comprising a computer controlled controller 140 (shown schematically in FIGS. 1-19). Controller 140 may be a control system or a computer connected to any of the devices of system 100, such as a separate computer, or it may be computer components integrated with one of the devices of system 100, such as an ASIC. These computer components may include one or more microprocessors, displays, keyboards (and/or other user input devices), memory components, printer(s), and the like. The controller 140 may be configured to receive signals from a user (eg, user commands) and/or from feedback devices such as pressure sensors, flow meters, and the like, and the ability to control the execution of fluid operations in the system 100. The controller 140 may contain software algorithms for implementing processes, such as the process using method 2000 depicted in FIG. 20 that allows the user to enter user-defined parameters associated with fluid operations in fluid device 102 of system 100, schedule various fluid operations in fluid device 102 of system 100, and/or cause controller 140 to perform various steps associated with fluid operations. liquids, control the execution of these operations and / or display their results (on a display or on a printer) for the user.

As shown in FIG. 1-19, controller 140 is in electrical communication with flow control valve 104, pump 106 (connection shown in broken lines), or intermediate devices configured to control flow control valve 104 and/or pump 106 (e.g., a stepper motor for valve 104 and motor for pump 106, etc.), so that controller 140 can instruct flow control valve 104 and pump 106 to perform various steps associated with operations in fluid handling processes (e.g., processes associated with FIGS. 2-19 and/or the method of Fig. 20). According to some examples, the controller 140 is configured to command the flow control valve 104 to fluidly connect a selected fluid reservoir from the reservoir set 110 to an inlet 103 such that fluid from the selected fluid reservoir can pass through the appropriate connection channel 131, the flow control valve 104, inlet 103, fluid device 102, outlet 105, and/or pump chamber 106. In some examples, controller 140 is configured to command pump 106 to move in a first or second direction to create a pressure difference between any one of the set of reservoirs. 110 tanks and an outlet channel 105 for pumping fluid towards the pump 106 or away from the pump 106.

According to some examples, the controller 140 is configured to access a computer-readable storage medium that contains computer-executable instruction codes for performing the various processes described above. In some examples, by executing instructions represented by codes in a computer readable medium, controller 140 causes system 100 to perform methods and processes, or parts thereof, including: (a) transferring an aliquot of the first liquid reactant to a fluidic device; (b) upon completion of the process (a) - moving the volume of the buffer solution into the fluid device; (c) upon completion of process (b), moving at least a portion of the volume of buffer solution transferred during process (b) into the reservoir of the first buffer solution; (d) upon completion of process (c), transferring an aliquot of the second liquid reactant to the fluid device; (e) upon completion of the process (d) - moving the volume of the buffer solution into the fluid device; and/or (f) upon completion of process (e), moving at least a portion of the volume of buffer solution displaced during process (e) into a reservoir of the second buffer solution.

It should be understood that all combinations of the above ideas and additional ideas, which will be discussed in more detail below (provided that these ideas do not contradict each other) are considered as part of the disclosed subject matter. In particular, all combinations of claimed subject matter appearing at the end of this description are considered part of the disclosed subject matter. It should also be understood that terms that are expressly used in the present description, and which may also appear in the description of any invention to which reference is made, should correspond to the meaning most appropriate to the specific ideas disclosed in this description.

Although the subject matter of the present invention has been described and presented in some detail with reference to certain illustrative examples, including various combinations and subcombinations of characteristics, the possibility of other examples, variations and modifications that fall within the scope of the present invention should be understood by those skilled in the art. Moreover, the descriptions of such examples, combinations and subcombinations are not intended to convey the idea that the claimed subject matter requires characteristics or combinations of characteristics other than those expressly listed in the claims. Accordingly, the scope of the present invention is intended to include all modifications and variations that fall within the scope of the appended claims.

Information disclosed in the invention includes the following items:

1. A method comprising:

(a) transferring an aliquot of the first liquid reactant to the fluid device;

(b) after step (a) - moving the volume of the buffer solution into the fluid device;

(c) after step (b), moving at least a portion of the volume of the buffer solution transferred in step (b) into the reservoir of the first buffer solution;

(d) after step (c), transferring an aliquot of the second liquid reactant to the fluid device;

(e) after step (d) - transferring the volume of the buffer solution into the fluid device;

(f) after step (e), transferring at least a portion of the volume of buffer solution transferred in step (e) into the second buffer reservoir.

2. The method according to claim 1, further comprising:

(g) after step (f), transferring an aliquot of the first liquid reactant to the fluid device;

(h) after step (g), transferring a volume of buffer solution to the fluid device, wherein at least a portion of the volume of buffer solution transferred in step (h) is transferred from the first buffer solution reservoir;

(i) after step (h), transferring an aliquot of the second liquid reactant to the fluid device; And

(j) after step (i), transferring the buffer solution volume to the fluid device, wherein at least a portion of the buffer solution volume transferred in step (j) is transferred from the second buffer solution reservoir.

3. The method according to claim 2, further comprising:

(k) after step (h) but before step (i), moving at least a portion of the volume transferred in step (h) back into the first buffer solution reservoir; And

(i) after step (j), moving at least a portion of the volume transferred in step (j) back to the second buffer solution reservoir.

4. The method according to any one of paragraphs. 1-3, wherein step (b) comprises transferring a buffer volume from the first buffer reservoir to the fluid device, and then transferring the buffer volume from the common buffer reservoir to the fluid device, wherein

step (e) comprises transferring a buffer volume from the second buffer reservoir to the fluid device and then transferring the buffer volume from the common buffer reservoir to the fluid device.

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

6. The method according to any one of paragraphs. 1-5, wherein step (b) comprises transferring a volume of buffer solution from the first buffer solution reservoir to the fluid device, wherein

step (e) comprises transferring a volume of buffer solution from the second buffer solution reservoir to the fluid device.

7. The method according to any one of paragraphs. 1-6, in which the proportion of the buffer solution transferred to the tank of the first buffer solution in step (c) is in the range of approximately 30% to 70% of the volume of the buffer solution transferred in step (b)

8. The method of claim 2, wherein in step (h) the proportion of buffer solution transferred from the first buffer reservoir is between about 30% and 70% of the volume of buffer solution transferred to the fluid device.

9. The method according to any one of paragraphs. 1-8, wherein in step (d) an aliquot of the second liquid reactant is not mixed with an aliquot of the first liquid reactant in the fluid device.

10. System containing:

fluid device;

flow control valve;

a first liquid reactant reservoir configured to be in fluid communication with the fluid device via a flow control valve;

a first buffer solution reservoir configured to be in fluid communication with the fluid device via a flow control valve;

a common source of buffer solution, made with the possibility of fluid communication with the fluid device through a flow control valve;

wherein the flow control valve allows for the following flows: (i) flow from the first liquid reactant reservoir to the fluid device; (ii) flow from the common buffer source to the fluid device; (iii) flow from the fluid device to the reservoir of the first buffer solution; (iv) flow from the first liquid reactant reservoir to the fluid device; and (v) flow from the first buffer solution reservoir to the fluid device.

11. The system according to claim 10, further comprising:

a second liquid reactant reservoir configured to be in fluid communication with the fluid device via a flow control valve;

a second buffer reservoir, configured to be in fluid communication with the fluid device via a flow control valve;

wherein the flow control valve further allows for the following fluid flows: (vi) flow from the second liquid reactant reservoir to the fluid device; (vii) flow from the second buffer reservoir to the fluid device; (viii) flow from the common buffer source to the fluid device; and (ix) flow from the fluid device to the second buffer reservoir.

12. The system of claim 10 or 11, wherein the first buffer solution reservoir comprises a cache channel that has a constant cross-sectional dimension along its length.

13. The system according to any one of paragraphs. 10-12, wherein the first buffer reservoir is in fluid communication with the fluid device via a channel, wherein the first buffer reservoir comprises a cache reservoir having a cross-sectional dimension larger than the channel cross-sectional dimension.

14. The system according to any one of paragraphs. 10-13 further comprising a syringe pump operatively associated with the fluid device, the first liquid reagent reservoir, the common buffer source, and the first buffer reservoir, the syringe pump configured to create a pressure difference between the fluid device and any one of the following: a first liquid reactant reservoir, a common buffer source, or a first buffer reservoir, to set the liquids in motion.

15. The system according to any one of paragraphs. 10-14 wherein the flow control valve is a rotary valve.

16. The system according to any one of paragraphs. 10-15, further comprising a controller that is in electrical communication with the flow control valve for transmitting commands that control the operation of the flow control valve in order to perform sequencing.

17. The system according to any one of paragraphs. 10-16, in which the reservoir of the first buffer solution contains a first volume of liquid, which is at least 30% of the volume of liquid that is contained in the fluid device.

18. A computer-readable medium that encodes computer-executable instructions that, when executed by the computer controller of the automated system, cause the system to perform the following processes:

(a) transferring an aliquot of the first liquid reactant to the fluid device;

(b) after process (a) - transferring a volume of buffer solution into the fluid device;

(c) after process (b), moving at least a portion of the volume of buffer solution transferred during process (b) into a reservoir of the first buffer solution;

(d) after process (c), transferring an aliquot of the second liquid reactant to the fluid device;

(e) after process (d) - transferring a volume of buffer solution to the fluid device; And

(f) after process (e), transferring at least a portion of the volume of buffer solution transferred during process (e) into a reservoir of the second buffer solution.

19. The computer-readable medium of claim 18, wherein the processes in the system further comprise:

(g) after process (f) - transferring an aliquot of the first liquid reactant to the fluid device;

(h) after process (g), transferring a volume of buffer solution into the fluid device, wherein at least a portion of the volume of buffer solution transferred during process (h) is transferred from the first buffer reservoir;

(i) after process (h) - transferring an aliquot of the second liquid reactant to the fluid device; And

(j) after process (i), transferring a volume of buffer into the fluid device, wherein at least a portion of the volume of buffer transferred in step (j) is transferred from the second buffer reservoir.

20. The computer-readable medium of claim 18 or 19, wherein the process (b) comprises moving a volume of buffer solution from the first buffer solution reservoir to the fluid device, and then moving the volume of buffer solution from the common buffer reservoir to the fluid device, wherein

process (e) comprises transferring a volume of buffer solution from the second buffer solution reservoir to the fluid device and then transferring the volume of buffer solution from the common buffer reservoir to the fluid device.

Claims (63)

1. A method for isolating and reusing a wash buffer solution, comprising: (a) moving (2010) an aliquot of the first liquid reagent (122) into the fluid device (102); (b) after step (a), moving (2015) a volume of buffer solution into the fluid device (102); (c) after step (b), moving (2020) at least part of the volume of the buffer solution (123) transferred in step (b) into the tank (113) of the first buffer solution; (d) after step (c), moving (2025) an aliquot of the second liquid reagent (124) into the fluid device (102); (e) after step (d) - moving (2030) the volume of the buffer solution into the fluid device (102); (f) after step (e), moving (2035) at least a portion of the volume of the buffer solution (124) transferred in step (e) into the tank (115) of the second buffer solution; (g) after step (f), moving (2040) an aliquot of the first liquid reactant (122) into the fluid device (102); (h) after step (g), moving (2045) the buffer volume into the fluid device (102), wherein at least a portion of the buffer volume transferred in step (h) is transferred from the first buffer reservoir (113). 2. The method according to p. 1, further comprising: (i) after step (h), moving (2050) an aliquot of the second liquid reactant (124) into the fluid device (102); And (j) after step (i), moving (2055) the buffer volume into the fluid device (102), wherein at least a portion of the buffer volume transferred in step (j) is transferred from the second buffer reservoir (115). 3. The method of claim 2, further comprising: (k) after step (h) but before step (i) - moving at least a portion of the volume transferred in step (h) back to the reservoir (113) of the first buffer solution ; And (l) after step (j), moving at least a portion of the volume transferred in step (j) back into the tank (115) of the second buffer solution. 4. The method according to any one of paragraphs. 1-3, wherein step (b) comprises moving the buffer volume from the first buffer reservoir (113) to the fluid device (102) and then transferring the buffer volume from the common buffer reservoir (111) to the fluid device (102) , wherein step (e) comprises transferring a buffer volume from the second buffer reservoir (115) to the fluid device (102), and then transferring the buffer volume from the common buffer reservoir (111) to the fluid device. 5. The method of claim 4, wherein in step (b) the volume of buffer solution transferred from the first buffer tank (113) is substantially equal to the volume of buffer solution transferred from the common buffer tank (111). 6. The method according to any one of paragraphs. 1-5, wherein step (b) comprises moving a volume of buffer solution from the first buffer reservoir (113) to the fluid device (102), wherein step (e) comprises transferring a volume of buffer solution from the second buffer reservoir (115) to the fluid device (102). 7. The method according to any one of paragraphs. 1-6, in which the proportion of the buffer solution transferred to the reservoir (113) of the first buffer solution in step (c) is in the range of approximately 30% to 70% of the volume of buffer solution transferred in step (b) 8. The method of claim 2, wherein in step (h) the proportion of buffer solution transferred from the first buffer reservoir (113) is in the range of approximately 30% to 70% of the volume of buffer solution transferred to the fluid device (102 ). 9. The method according to any one of paragraphs. 1-8, wherein in step (d) an aliquot of the second reactant liquid (124) is not mixed with an aliquot of the first reactant liquid (123) in the fluid device (102). 10. A system for isolating and reusing a flushing buffer, comprising: fluid device (102); flow control valve (104); a reservoir (112) of the first liquid reactant configured to be fluidly connected to the fluid device (102) via a flow control valve (104); a reservoir (113) of the first buffer solution, made with the possibility of fluid connection with the fluid device (102) through the valve (104) flow control; a source (111) of a common buffer solution, made with the possibility of fluid connection with the fluid device (102) through the valve (104) flow control; And a pump (106) operatively connected to the fluid device (102), the first liquid reactant reservoir (112), the common buffer source (111), and the first buffer reservoir (113); while the flow control valve (104) and the pump (106) are configured to sequentially provide: (i) flow from the first liquid reactant reservoir to the fluid device; (ii) flow from a common source of buffer solution to the fluid device; (iii) flow from the fluid device to the reservoir of the first buffer solution; (iv) flow from the first liquid reactant reservoir to the fluid device; (v) flow from the first buffer solution reservoir to the fluid device. 11. The system according to claim 10, further comprising: a reservoir (114) of the second liquid reagent, made with the possibility of liquid connection with the fluid device (102) through the valve (104) flow control; a tank (115) of the second buffer solution, made with the possibility of fluid connection with the fluid device (102) through the valve (104) flow control; wherein the flow control valve (104) and the pump (106) are further configured to provide after (i)-(iii) but before (iv)-v): (vi) flow from the second liquid reactant reservoir to the fluid device; (vii) flow from the second buffer solution reservoir to the fluid device; (viii) flow from the common buffer solution source to the fluid device; And (ix) flow from the fluid device to the second buffer reservoir. 12. The system according to claim 10 or 11, wherein the reservoir (113) of the first buffer solution comprises a cache channel which has a constant cross-sectional dimension along its length. 13. The system according to any one of paragraphs. 10-12, in which the reservoir (113) of the first buffer solution has a fluid connection with the fluid device (102) through a channel, and the tank of the first buffer solution contains a cache tank, the size of the cross-section of which is larger than the cross-sectional size of the specified channel. 14. The system according to any one of paragraphs. 10-13, wherein said pump is a syringe type pump configured to create a pressure differential between the fluid device (102) and any one of the following: a first liquid reactant reservoir (112), a common buffer source (111), or reservoir (113) the first buffer solution, in order to set the liquids in motion. 15. The system according to any one of paragraphs. 10-14, in which the flow control valve (104) is a rotary valve. 16. The system according to any one of paragraphs. 10-15, further comprising a controller (140) that is in electrical communication with the flow control valve (104) for transmitting commands controlling the operation of the flow control valve (104) in order to perform sequencing. 17. The system according to any one of paragraphs. 10-16, in which the reservoir of the first buffer solution contains a first volume of liquid, which is at least 30% of the volume of liquid that is contained in the fluid device. 18. A machine-readable medium for isolating and reusing the wash buffer solution, which encodes computer-executable instructions that, when executed by the computer controller of the automated system, cause the system to perform the following processes: (a) transferring (2010) an aliquot of the first liquid reactant to the fluid device; (b) after process (a) - moving (2015) a volume of the buffer solution into the fluid device; (c) after process (b), moving (2020) at least a portion of the volume of buffer solution transferred during process (b) into the first buffer tank; (d) after process (c), transferring (2025) an aliquot of the second liquid reactant to the fluid device; (e) after process (d) - moving (2030) a volume of buffer solution into the fluid device; And (f) after process (e), moving (2035) at least a portion of the volume of buffer solution transferred during process (e) into a second buffer tank; (g) after process (f) - moving (2040) an aliquot of the first liquid reactant into the fluid device; (h) after process (g), moving (2045) a volume of buffer solution into the fluid device, wherein at least a portion of the volume of buffer solution transferred during process (h) is transferred from the first buffer reservoir. 19. The computer-readable medium of claim 18, wherein the processes in the system further comprise: (i) after process (h) - moving (2050) an aliquot of the second liquid reactant into the fluid device; And (j) after process (i), moving (2055) a volume of buffer solution into the fluid device, wherein at least a portion of the volume of buffer solution transferred in step (j) is transferred from the second buffer reservoir. 20. The computer-readable medium of claim 18 or 19, wherein the process (b) comprises moving a volume of buffer solution from the first buffer solution reservoir to the fluid device, and then moving the volume of buffer solution from the common buffer reservoir to the fluid device, wherein process (e) comprises transferring a volume of buffer solution from the second buffer solution reservoir to the fluid device and then transferring the volume of buffer solution from the common buffer reservoir to the fluid device.