Patent Description:
<CIT> discloses a system and a method for generating samples for biochemical analysis and/or conducting biochemical reactions. The system comprises a fluidic network comprising a sample channel, a reaction chamber, and a reservoir, the sample channel being in flow communication with a sample port that is to receive a biological sample; a pump assembly in flow communication with the fluidic network; and a rotary valve comprising a flow channel and rotatable between first and second valve positions.

<CIT> concerns a flow cell liquid degassing system and method.

<CIT> describes a method and a device for performing molecular reactions.

<CIT> discloses a fluidic system that includes a reagent manifold comprising channels configured for fluid communication between a reagent cartridge and an inlet of a flow cell; reagent sippers extending downward from ports in the manifold, each of the reagent sippers configured to be placed into a reagent reservoir in a reagent cartridge so that liquid reagent can be drawn from the reagent reservoir into the sipper; and at least one valve configured to mediate fluid communication between the reservoirs and the inlet of the flow cell.

<CIT> concerns apparatus and methods for fluidic manipulation and optical detection of samples.

The following presents a simplified summary in order to provide a basic understanding of some aspects described herein. This summary is not an extensive overview of the claimed subject matter. It is intended to neither identify key or critical elements of the claimed subject matter nor delineate the scope thereof. Its sole purpose is to present some concepts in a simplified form as a prelude to the more detailed description that is presented later.

In a first aspect of the invention, a method as defined in claim <NUM> is provided.

In a second aspect of the invention, a system as defined in claim <NUM> is provided.

In a third aspect of the invention, a computer readable medium encoded with computer-executable instructions as defined in claim <NUM> is provided.

Other features and characteristics of the subject matter of this disclosure, as reservoir as the methods of operation, functions of related elements of structure and the combination of parts, and economies of manufacture, will become more apparent upon consideration of the following description and the appended claims with reference to the accompanying drawings, all of which form a part of this specification, wherein like reference numerals designate corresponding parts in the various figures.

The accompanying drawings, which are incorporated herein and form part of the specification, illustrate various examples of the subject matter of this disclosure. In the drawings, like reference numbers indicate identical or functionally similar elements.

While aspects of the subject matter of the present disclosure may be embodied in a variety of forms, the following description and accompanying drawings are merely intended to disclose some of these forms as specific examples of the subject matter. Accordingly, the subject matter of this disclosure is not intended to be limited to the forms or examples so described and illustrated.

Unless defined otherwise, all terms of art, notations and other technical terms or terminology used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs.

Unless otherwise indicated or the context suggests otherwise, as used herein, "a" or "an" means "at least one" or "one or more.

This description may use relative spatial and/or orientation terms in describing the position and/or orientation of a component, apparatus, location, feature, or a portion thereof. Unless specifically stated, or otherwise dictated by the context of the description, such terms, including, without limitation, top, bottom, above, below, under, on top of, upper, lower, left of, right of, in front of, behind, next to, adjacent, between, horizontal, vertical, diagonal, longitudinal, transverse, radial, axial, etc., are used for convenience in referring to such component, apparatus, location, feature, or a portion thereof in the drawings and are not intended to be limiting.

Furthermore, unless otherwise stated, any specific dimensions mentioned in this description are merely representative of an example implementation of a device embodying aspects of the disclosure and are not intended to be limiting.

The use of the term "about" applies to all numeric values specified herein, whether or not explicitly indicated. This term generally refers to a range of numbers that one of ordinary skill in the art would consider as a reasonable amount of deviation to the recited numeric values (i.e., having the equivalent function or result) in the context of the present disclosure. For example, and not intended to be limiting, this term can be construed as including a deviation of ±<NUM> percent of the given numeric value provided such a deviation does not alter the end function or result of the value. Therefore, under some circumstances as would be appreciated by one of ordinary skill in the art a value of about <NUM>% can be construed to be a range from <NUM>% to <NUM>%.

As used herein, the term "adjacent" refers to being near or adjoining. Adjacent objects can be spaced apart from one another or can be in actual or direct contact with one another. In some instances, adjacent objects can be coupled to one another or can be formed integrally with one another.

As used herein, the terms "substantially" and "substantial" refer to a considerable degree or extent. When used in conjunction with, for example, an event, circumstance, characteristic, or property, the terms can refer to instances in which the event, circumstance, characteristic, or property occurs precisely as reservoir as instances in which the event, circumstance, characteristic, or property occurs to a close approximation, such as accounting for typical tolerance levels or variability of the examples described herein.

As used herein, the terms "optional" and "optionally" mean that the subsequently described, component, structure, element, event, circumstance, characteristic, property, etc. may or may not be included or occur and that the description includes instances where the component, structure, element, event, circumstance, characteristic, property, etc. is included or occurs and instances in which it is not or does not.

A "reagent" as used herein refers to any substance or combination thereof that participates in a molecular assay, other than sample material and products of the assay. Exemplary reagents include nucleotides, enzymes, amplification oligomers, probes, and salts.

The term "buffer" as used herein refers to any solution with a controlled pH that may serve to dissolve a solid (e.g., lyophilized) substance (e.g., reagent, sample, or combination thereof) or as a diluent to dilute a liquid (e.g., a liquid reagent, liquid sample, or combination thereof; or a solution of a reagent, sample, or combination thereof).

According to various examples, assemblies and devices as described herein may be used in combination with a fluid cartridge that may comprise one or more fluid processing passageways including one or more elements, for example, one or more of a channel, a branch channel, a valve, a flow splitter, a vent, a port, an access area, a via, a bead, a reagent containing bead, a cover layer, a reaction component, any combination thereof, and the like. Any element may be in fluid communication with another element.

All possible combinations of elements and components described in the specification or recited in the claims are contemplated and considered to be part of this disclosure. It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in greater detail below (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein, insofar as they fall under the scope of the appended claims.

In the appended claims, the term "including" is used as the plain-English equivalent of the respective term "comprising. " The terms "comprising" and "including" are intended herein to be open-ended, including not only the recited elements, but further encompassing any additional elements.

The term "fluid communication" means either direct fluid communication, for example, two regions can be in fluid communication with each other via an unobstructed fluid processing passageway connecting the two regions or can be capable of being in fluid communication, for example, two regions can be capable of fluid communication with each other when they are connected via a fluid processing passageway that can comprise a valve disposed therein, wherein fluid communication can be established between the two regions upon actuating the valve, for example, by dissolving a dissolvable valve, bursting a burstable valve, or otherwise opening a valve disposed in the fluid processing passageway.

While reusing fluid buffer after being flushed through the fluidic device curtails the amount of fluid buffer needed for a fluid sequence process, used fluid buffer typically includes remnants of the active reagent since sequencing procedures can instruct fluid handling equipment to introduce fluid buffers into the fluidic device directly after introducing the active reagent through the fluidic device. Consequently, mixing reused fluid buffer with other fluid operations of the sequence process can further contaminate the fluid buffer supply to the fluidic device and/or compromise the integrity of the fluid operations by inadvertently introducing remnant reagents from a prior fluid operation from the reused fluid buffer.

Thus, there is a need for improved fluidic systems and methods that allow fluid buffer to be both sequestered and reused in a sequencing process to avoid contamination and reduce the volume of fluid buffer needed to be stored by the fluidic cartridge to conduct a fluid sequence process.

Accordingly, the system is configured to sequester and reuse at least first and second reagent fluids and fluid buffers directed through a fluidic device by comprising one common fluid buffer reservoir to hold fresh, unused fluid buffer and at least first and second dedicated fluid buffer reservoirs dedicated to holding fluid buffer that has been used to flush each of the at least first and second different reagent fluids through the fluidic device. The system further comprises a flow control valve operatively associated with the common fluid buffer and the at least first and second dedicated fluid buffer reservoirs to selectively fluidly connect any one of the common fluid buffer reservoir and the at least first and second dedicated fluid buffer reservoirs to the fluidic device during a wash operation. Accordingly, by sequestering and reusing fluid buffer through the use of at least first and second dedicated fluid buffer reservoirs, the system minimizes the volume of fluid buffer used in a fluid process, such as a fluid sequencing operation, for example a sequencing-by-synthesis (SBS) operation that includes a cleave process, a scavenger process, an incorporation process, and a scan process interposed by one or more wash operations.

<FIG> show an example system <NUM> for sequestering and reusing reagent fluid and fluid buffer during two or more fluid operations. In some examples, the system <NUM> comprises a fluidic device <NUM>, an inlet channel <NUM>, a flow control valve <NUM>, an outlet channel <NUM>, a pump <NUM>, a set of fluid reservoirs <NUM>, and a set of connecting channels <NUM>. In some examples, the system <NUM> is part of a fluid cartridge (not shown) supporting various components of the system <NUM>, such as, the fluidic device <NUM>, the flow control valve <NUM>, the pump <NUM>, and the set of fluid reservoirs <NUM>, although one or more components of the system <NUM> may not be supported on a common fluid cartridge or other supporting structure.

As shown in <FIG>, the fluidic device <NUM> (e.g., flow cell) is fluidly connected to the flow control valve <NUM> by the inlet channel <NUM>. In one example, the fluidic device <NUM> is a flow cell comprising a first glass layer (not shown) and a second glass layer (not shown) secured together and defining one or more channels (not shown) therein that can be fluidically manipulated and optically detected. In various examples, the fluidic device <NUM> may include a fluid inlet <NUM>, a fluid outlet <NUM>, and one or more fluid channels (not shown) are fluidly connected to the fluid inlet and the fluid outlet to allow fluid processing, such as a chemical or biochemical assay or other process or reaction, to take place. In various examples, the fluidic device <NUM> is configured to allow the introduction of various types of fluids (e.g., reagents, buffers, reaction media) into the fluid inlet <NUM> to undergo fluid processing within the one or more fluid channels. In various examples, fluidic device <NUM> is further configured to allow the various types of fluids to be flushed out of the one or more fluid channels through the fluid outlet <NUM>.

In the example shown in <FIG>, the inlet channel <NUM> fluidly connects the fluid inlet <NUM> of the fluidic device <NUM> to the flow control valve <NUM>, and the outlet channel <NUM> fluidly connects the fluid outlet <NUM> of the fluidic device <NUM> to the pump <NUM>. In other examples (not shown), the system <NUM> may include two or more channels to fluidly connect the fluidic device <NUM> to the flow control valve <NUM> and two or more channels to fluidly connect the fluidic device <NUM> to the pump <NUM>.

The set of fluid reservoirs <NUM> comprises two or more reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM>. In the example shown in <FIG>, the two or more reagent fluid reservoirs include a first reagent fluid reservoir <NUM>, a second reagent fluid reservoir <NUM>, a third reagent fluid reservoir <NUM>, and a fourth reagent fluid reservoir <NUM>, although any number of reagent fluid reservoirs is contemplated by this disclosure. The different reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> of the set of fluid reservoirs <NUM> may have the same or varying sizes (i.e., volumes) - e.g., all reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and <NUM> may have the same volume, all reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and <NUM> may have different volumes, or a subset of the reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have the same volume and a subset of the reagent reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have different volumes - depending on the necessary storage volume of the reagent to be stored in each reagent fluid reservoir.

In some examples, the first reagent fluid reservoir <NUM> holds a first reagent fluid <NUM> comprising a solvent and a first reagent directed to a first reagent operation (e.g., cleave). In some examples, the second reagent fluid reservoir <NUM> holds a second reagent fluid <NUM> comprising a solvent and a second reagent directed to a second reagent operation (e.g., scan). In some examples, the third reagent fluid reservoir <NUM> holds a third reagent fluid <NUM> comprising a solvent and a third reagent directed to a third reagent operation (e.g., incorporation). In some examples, the fourth reagent fluid reservoir <NUM> holds a fourth reagent fluid <NUM> comprising a solvent and a fourth reagent directed to a fourth reagent operation (e.g., scavenger).

The set of fluid reservoirs <NUM> comprises three or more fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM>. In the example shown in <FIG>, the three or more fluid buffer reservoirs include a common fluid buffer reservoir <NUM> and at least two dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM>, each associated with one of the reagent fluids <NUM>, <NUM>, <NUM>, and/or <NUM>. In the examples shown in <FIG>, the at least two dedicated fluid buffer reservoirs comprise a first dedicated fluid buffer reservoir <NUM>, a second dedicated fluid buffer reservoir <NUM>, and/or a third fluid buffer reservoir <NUM>, although any number of dedicated fluid buffer reservoirs is contemplated by this disclosure. The different fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> of the set of reservoirs <NUM> may have the same or varying sizes (i.e., volumes) - e.g., all fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have the same volume, all fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have different volumes, or a subset of the fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have the same volume and a subset of the fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> may have different volumes - depending on the necessary storage volume of the reagent to be stored in each reagent fluid reservoir.

In some examples, each of the dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM> holds a volume of fluid that is at least <NUM>% of a volume of fluid held by the fluidic device <NUM>. In some examples, each of the dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM> is a cache channel comprising a consistent cross-sectional dimension across a length thereof. In some examples, each of the dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM> is a cache well comprising a cross-sectional dimension larger than a cross-sectional dimension of its associated connecting channel <NUM>. In some examples, the cache-well does not include any sharp edges or various topographical features and is configured to minimize bubble nucleation such that the cache-well does not accumulate bubbles as fluid is pushed in and out of the reservoir.

In the examples shown in <FIG>, the common fluid buffer reservoir <NUM> holds a common fluid buffer <NUM> (e.g., wash solution that includes salt-water and soap) that has not been flushed through the fluidic device <NUM>. In various examples, each of the dedicated buffer reservoirs <NUM>, <NUM>, and/or <NUM> holds a sequestered fluid buffer that has been flushed through the fluidic device <NUM> following a reagent operation conducted in the fluidic device <NUM>, e.g., with one of the reagent fluids <NUM>, <NUM>, and/or <NUM>, although initially before any reagent operations have occurred, the dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM> may hold unused fluid buffer.

In the examples shown in <FIG>, the first buffer reservoir <NUM> holds a first fluid buffer <NUM> that includes a mixture of used fluid buffer <NUM> that has been flushed through the fluidic device <NUM> after the first reagent fluid <NUM> has been directed through the fluidic device <NUM> during the first reagent operation. In the examples shown in <FIG>, the second buffer reservoir <NUM> holds a second fluid buffer <NUM> that includes a mixture of used fluid buffer <NUM> that has been flushed through the fluidic device <NUM> after the second reagent fluid <NUM> has been directed through the fluidic device <NUM> during the second reagent operation. In the examples shown in <FIG>, the third buffer reservoir <NUM> holds a third fluid buffer <NUM> that includes a mixture of used fluid buffer <NUM> that has been flushed through the fluidic device <NUM> after the third reagent fluid <NUM> has been directed through the fluidic device <NUM> during the fourth reagent operation.

In some examples, the system may not include a dedicated fluid buffer reservoir for every reagent. For example, system and processes shown in <FIG> do not include a dedicated fluid buffer reservoir associated with the fourth reagent fluid <NUM>, other example systems and processes may include a dedicated fluid buffer reservoir associated with every reagent fluid.

In the example shown in <FIG>, the set of connecting channels <NUM> comprises a respective connecting channel <NUM> extending from its associated fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> to the flow control valve <NUM>, such that the flow control valve <NUM> is fluidly connected to each fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of the set of fluid reservoirs <NUM>. In other examples (not shown), the set of connecting channels <NUM> may include two or more channels to fluidly connect a respective fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> to the flow control valve <NUM>, such as a front connecting channel (such as the respective connecting channel <NUM> shown) and a rear connecting channel (not shown) such that the used fluid buffer can be recycled to the rear of the respective fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>, opposite the front connecting channel.

Flow control valve <NUM> is constructed and arranged to selectively, fluidly connect one of the fluid reservoirs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of the set of fluid reservoirs <NUM> to the inlet channel <NUM>, and thus, to the fluidic device <NUM>. According to various examples, the flow control valve <NUM> comprises a rotary valve for selectively connecting to one of the connecting channels <NUM> for a respective fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>.

In the example shown in <FIG>, the flow control valve <NUM> is a rotary valve comprising a rotary body <NUM> and valve selector channel <NUM>. In some examples, the rotary body <NUM> is configured to rotate between a plurality of angular positions so that the valve selector channel <NUM> may fluidly connect any one of the fluid reservoirs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> to a valve outlet port via a respective inlet port for each fluid reservoir. When the rotary body <NUM> is rotated to an angular position such that the valve selector channel <NUM> is aligned with the one of the inlet ports for a selected fluid reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>, fluid may flow from the selected reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>, through the valve selector channel <NUM>, and into the valve outlet port.

In other examples (not shown), the flow control valve <NUM> may include any other type of valve to selectively, fluidly connect one of the fluid reservoirs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> to the inlet channel <NUM>. In other examples (not shown), the flow control valve <NUM> may include a valve array, such as a plurality of pinch valves or solenoid valves and a manifold, to selectively, fluidly connect one of the fluid reservoirs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> to the inlet channel <NUM>.

In various examples, the pump <NUM>, fluidly connected to the outlet channel <NUM>, is configured to apply a pressure differential between any one <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of the first set of fluid reservoirs <NUM> and the outlet channel <NUM> to propel fluid flow bidirectionally along a respective connecting channel <NUM> of the set of connecting channels <NUM>, the flow control valve <NUM>, inlet channel <NUM>, the fluidic device <NUM>, and the outlet channel <NUM>. Pump <NUM> may comprise a syringe pump with an actuator (not shown) operatively associated with the syringe. In various examples, the actuator is configured to move a plunger of the syringe in a first direction to generate a negative pressure differential to draw fluid through the fluidic device <NUM> toward (and possibly into) a barrel of the syringe. The actuator is further configured to move the plunger in a second direction, opposite to the first direction, to generate a positive pressure differential and expel fluid away from (and possible out of) the syringe toward a selected reservoir <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> of the set of fluid reservoirs <NUM>. Accordingly, by moving in a second direction to generate a positive pressure differential, the pump <NUM> is configured to expel fluid held in the fluidic device <NUM> or the outlet channel <NUM> though the inlet channel <NUM>, the flow control valve <NUM>, a selected connecting channel <NUM>, and into one of the selected fluid reservoirs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or <NUM>.

In other examples (not shown), the pump <NUM> may comprise any other pressure differential creating mechanism that is capable of reversing flow direction.

In various examples, as shown in <FIG>, the system <NUM> sequesters and reuses reagent fluids and fluid buffers by: (i) selectively directing reagent fluids from any one of the reagent fluid reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> to the fluidic device <NUM> to perform a reagent operation; (ii), optionally, redirecting at least a portion of used reagent fluid <NUM>, <NUM>, <NUM>, and/or <NUM> directed through the fluidic device <NUM> to the selected reagent fluid reservoir <NUM>, <NUM>, <NUM>, and <NUM> to be reused for a subsequent reagent operation; (iii) selectively directing fluid buffers from any of the fluid buffer reservoirs <NUM>, <NUM>, <NUM>, and/or <NUM> to the fluidic device <NUM> to conduct a wash operation; and (iv) redirecting at least a portion of a used fluid buffer <NUM>, <NUM>, and/or <NUM> directed through the fluidic device <NUM> back to one of the dedicated fluid buffer reservoirs <NUM>, <NUM>, and/or <NUM> to be reused for a subsequent wash operation.

Referring to <FIG>, the system <NUM> may be set to perform a first reagent operation, such that flow control valve <NUM> permits fluid flow from the selected first reagent fluid reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>. During the first reagent operation, the flow control valve <NUM> is set to connect the first reagent fluid reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first reagent fluid reservoir <NUM>). The pump <NUM> is operated to draw the first reagent fluid <NUM> from the first reagent fluid reservoir <NUM> through the flow control valve <NUM> and into the fluidic device <NUM>. As shown in <FIG>, an aliquot <NUM> of the first reagent fluid <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a first reagent reverse operation, such that the pump <NUM> redirects at least a portion of the aliquot <NUM> of the first reagent fluid <NUM> moved in the first reagent operation back to the first reagent fluid reservoir <NUM>. During the first reagent reverse operation, the flow control valve <NUM> remains set to connect the fluidic device <NUM> to the first reagent fluid reservoir <NUM>, and the pump <NUM> is operated to expel fluid in a reverse direction through the fluidic device <NUM> back into the first reagent fluid reservoir <NUM>. As shown in <FIG>, at least a portion <NUM> of the aliquot <NUM> of the first reagent fluid <NUM> moved in the first reagent operation is received back in the first reagent fluid reservoir <NUM> to be reused for one or more subsequent first reagent operations.

Referring to <FIG> and <FIG>, the system <NUM> may be set to perform a first wash operation such that flow control valve <NUM> initially permits fluid flow from the selected first dedicated fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>, and then, optionally, permits fluid flow from the common fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>.

As shown in <FIG>, during a first part of the first wash operation, the flow control valve <NUM> is set to connect the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the first fluid buffer <NUM> from the first dedicated fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, a first volume <NUM> of the first fluid buffer <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM> to flush the first reagent fluid <NUM> remaining in the fluidic device <NUM>. In some implementations, the first volume <NUM> may include a volume <NUM> of reused first fluid buffer, shown in <FIG>, in instances where the first fluid buffer <NUM> has previously been pumped into the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>.

As shown in <FIG>, in some examples, during a second part of the first wash operation, after the first volume <NUM> of the first fluid buffer <NUM> is flushed through the fluidic device <NUM>, the flow control valve <NUM> is set to connect the common fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, a second volume <NUM> of the common fluid buffer <NUM> can be moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>. In some implementations, the second volume <NUM> can mix with the first volume <NUM> in one or more of the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>. Thus, if the first volume <NUM> includes a volume <NUM> of reused first fluid buffer, shown and described in <FIG>, the inclusion of the common fluid buffer <NUM> can dilute or otherwise lessen the presence of remnant first reagent fluid <NUM> in the reused first fluid buffer. In addition, by providing the common fluid buffer <NUM> after the reused first fluid buffer, the common fluid buffer <NUM> can fluidically separate the reused first fluid buffer further downstream from the fluidic device <NUM>.

In other examples, the second part of the first wash operation illustrated in <FIG> may be omitted and only the first fluid buffer <NUM> for the first wash operation can be drawn from the first dedicated fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a first buffer reverse operation such that flow control valve <NUM> permits fluid flow to the first dedicated fluid buffer reservoir <NUM>. The pump <NUM> redirects a third volume <NUM> of used fluid buffer comprising the first fluid buffer <NUM> and/or the common fluid buffer <NUM> back to the first dedicated fluid buffer reservoir <NUM>. The third volume <NUM> can include at least a portion of remnant first reagent fluid <NUM> in addition to the first fluid buffer <NUM> and/or the common fluid buffer <NUM>. In some instances, the percentage of the common fluid buffer <NUM> in the third volume <NUM> is greater than a percentage of first fluid buffer <NUM> and/or reused first fluid buffer. During the first buffer reverse operation, the flow control valve <NUM> is set to connect the fluidic device <NUM> to the first dedicated fluid buffer reservoir <NUM>(e.g., by connecting valve selector channel <NUM> with the connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>), and the pump <NUM> is operated to move fluid in a reverse direction through the fluidic device <NUM> to the first dedicated fluid buffer reservoir <NUM>. As shown in <FIG>, the third volume <NUM> of used fluid buffer is received back in the first dedicated fluid buffer reservoir <NUM> to be reused for one or more subsequent first wash operations. In some examples, the third volume <NUM> of used fluid buffer is less than or equal to the second volume <NUM> moved from the common fluid buffer reservoir <NUM>, in other examples, the third volume <NUM> of used fluid buffer is greater than the second volume <NUM> moved from the common fluid buffer reservoir <NUM>, and, in other examples, the third volume <NUM> of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device <NUM>, i.e., the combined first fluid volume <NUM> moved from the first fluid dedicated buffer reservoir <NUM> and second volume <NUM> moved from the common fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a second reagent operation, such that flow control valve <NUM> permits fluid flow from the selected second reagent fluid reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>. During the second reagent operation, the flow control valve <NUM> is set to connect the second reagent fluid reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the second reagent fluid reservoir <NUM>). The pump <NUM> is operated to draw the second reagent fluid <NUM> from the second reagent fluid reservoir <NUM> through the flow control valve <NUM> and into the fluidic device <NUM>. As shown in <FIG>, an aliquot <NUM> of the second reagent fluid <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a second reagent reverse operation, such that the pump <NUM> redirects at least a portion of the aliquot <NUM> of the second reagent fluid <NUM> moved in the second reagent operation back to the second reagent fluid reservoir <NUM>. During the second reagent reverse operation, the flow control valve <NUM> remains set to connect the fluidic device <NUM> to the second reagent fluid reservoir <NUM>, and the pump <NUM> is operated to expel fluid in a reverse direction through the fluidic device <NUM> back into the second reagent fluid reservoir <NUM>. As shown in <FIG>, at least a portion <NUM> of the aliquot <NUM> of the second reagent fluid <NUM> moved in the second reagent operation is received back in the second reagent fluid reservoir <NUM> to be reused in one or more subsequent second reagent operations.

Referring to <FIG> and <FIG>, the system <NUM> may be set to perform a second wash operation such that flow control valve <NUM> initially permits fluid flow from the selected second dedicated fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>, and then, optionally, permits fluid flow from the common fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>.

As shown in <FIG>, during a first part of the second wash operation, the flow control valve <NUM> is set to connect the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the second buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the second fluid buffer <NUM> from the second dedicated fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, a fourth volume <NUM> of the second fluid buffer <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM> to flush the second reagent fluid <NUM> remaining in the fluidic device <NUM>. In some implementations, the fourth volume <NUM> may include a volume <NUM> of reused second fluid buffer, shown in <FIG>, in instances where the second fluid buffer <NUM> has previously been pumped into the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>.

As shown in <FIG>, in some examples, during a second part of the second wash operation, after the fourth volume <NUM> of the second fluid buffer <NUM> is flushed through the fluidic device <NUM>, the flow control valve <NUM> is set to connect the common fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, a fifth volume <NUM> of the common fluid buffer <NUM> can be moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>. In some implementations, the fifth volume <NUM> can mix with the fourth volume <NUM> in one or more of the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>. Thus, if the fourth volume <NUM> includes a volume <NUM> of reused second fluid buffer, shown and described in <FIG>, the inclusion of the common fluid buffer <NUM> can dilute or otherwise lessen the presence of remnant second reagent fluid <NUM> in the reused second fluid buffer. In addition, by providing the common fluid buffer <NUM> after the reused second fluid buffer, the common fluid buffer <NUM> can fluidically separate the reused second fluid buffer further downstream from the fluidic device <NUM>.

In other examples, the second part of the second wash operation illustrated in <FIG> may be omitted and only the second fluid buffer <NUM> for second wash operation can be drawn from the second dedicated fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a second buffer reverse operation such that flow control valve <NUM> permits fluid flow to the second dedicated fluid buffer reservoir <NUM>. The pump <NUM> redirects a sixth volume <NUM> of used fluid buffer comprising the second fluid buffer <NUM> and/or the common fluid buffer <NUM> back to the second dedicated fluid buffer reservoir <NUM>. The sixth volume <NUM> can include at least a portion of remnant second reagent fluid <NUM> in addition to the second fluid buffer <NUM> and/or the common fluid buffer <NUM>. In some instances, the percentage of the common fluid buffer <NUM> in the sixth volume <NUM> is greater than a percentage of second fluid buffer <NUM> and/or reused second fluid buffer. During the second buffer reverse operation, the flow control valve <NUM> is set to connect the fluidic device <NUM> to the second dedicated fluid buffer reservoir <NUM> (e.g., by connecting valve selector channel <NUM> with the connecting channel <NUM> associated with the second dedicated fluid buffer reservoir <NUM>), and the pump <NUM> is operated to move fluid in a reverse direction through the fluidic device <NUM> to the second dedicated fluid buffer reservoir <NUM>. As shown in <FIG>, the sixth volume <NUM> of used fluid buffer is received back in the second dedicated fluid buffer reservoir <NUM> to be reused for one or more subsequent second wash operations. In some examples, the sixth volume <NUM> of used fluid buffer is less than or equal to the fifth volume <NUM> moved from the common fluid buffer reservoir <NUM>, in other examples, the sixth volume <NUM> of used fluid buffer is greater than the fifth volume <NUM> moved from the common fluid buffer reservoir <NUM>, and, in other examples, the sixth volume <NUM> of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device <NUM>, i.e., the combined fourth volume <NUM> moved from the second dedicated fluid buffer reservoir <NUM> and fifth volume <NUM> moved from the common fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a third reagent operation, such that flow control valve <NUM> permits fluid flow from the selected third reagent fluid reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>. During the third reagent operation, the flow control valve <NUM> is set to connect the third reagent fluid reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the third reagent fluid reservoir <NUM>). The pump <NUM> is operated to draw the third reagent fluid <NUM> from the third reagent fluid reservoir <NUM> through the flow control valve <NUM> and into the fluidic device <NUM>. As shown in <FIG>, an aliquot <NUM> of the third reagent fluid <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a third reagent reverse operation, such that the pump <NUM> redirects at least a portion of the aliquot <NUM> of the third reagent fluid <NUM> moved in the third reagent operation back into the third reagent fluid reservoir <NUM>. During the third reagent reverse operation, the flow control valve <NUM> remains set to connect the fluidic device <NUM> to the third reagent fluid reservoir <NUM>, and the pump <NUM> is operated to expel fluid in a reverse direction through the fluidic device <NUM> to the third reagent fluid reservoir <NUM>. As shown in <FIG>, at least a portion <NUM> of the aliquot <NUM> of the third reagent fluid <NUM> moved in the third reagent operation is received back in the third reagent fluid reservoir <NUM> to be reused for one or more subsequent third reagent operations.

Referring to <FIG> and <FIG>, the system <NUM> may be set to perform a third wash operation, such that flow control valve <NUM> initially permits fluid flow from the selected third fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>, and then, optionally, permits fluid flow from the common fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>.

As shown in <FIG>, during a first part of the third wash operation, the flow control valve <NUM> is set to connect the third fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the third buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the third fluid buffer <NUM> from the third fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, a seventh volume <NUM> of the third fluid buffer <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM> to flush the third reagent fluid <NUM> remaining in the fluidic device <NUM>. In some implementations, the seventh volume <NUM> may include a volume <NUM> of reused third fluid buffer, shown in <FIG>, in instances where the third fluid buffer <NUM> has previously been pumped into the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>.

As shown in <FIG>, in some examples, during a second part of the third wash operation, after the seventh volume <NUM> of the third fluid buffer <NUM> is flushed through the fluidic device <NUM>, the flow control valve <NUM> is set to connect the common fluid buffer reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>). Pump <NUM> is operated to draw an aliquot of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> through the fluidic device <NUM>. As shown in <FIG>, an eighth volume <NUM> of the common fluid buffer <NUM> can be moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>. In some implementations, the eight volume <NUM> can mix with the seventh volume <NUM> in one or more of the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>. Thus, if the seventh volume <NUM> includes a volume <NUM> of reused third fluid buffer, shown and described in <FIG>, the inclusion of the common fluid buffer <NUM> can dilute or otherwise lessen the presence of remnant third reagent fluid <NUM> in the reused third fluid buffer. In addition, by providing the common fluid buffer <NUM> after the reused third fluid buffer, the common fluid buffer <NUM> can fluidically separate the reused third fluid buffer further downstream from the fluidic device <NUM>.

In other examples, the second part of the second wash operation illustrated in <FIG> may be omitted and only the third fluid buffer <NUM> for third wash operation can be drawn from the third fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a third buffer reverse operation such that flow control valve <NUM> permits fluid flow to the third fluid buffer reservoir <NUM>. The pump <NUM> redirects a ninth volume <NUM> of used fluid buffer comprising the third fluid buffer <NUM> and/or the common fluid buffer <NUM> back to the third fluid buffer reservoir <NUM>. The ninth volume <NUM> can include at least a portion of remnant third reagent fluid <NUM> in addition to the third fluid buffer <NUM> and/or the common fluid buffer <NUM>. In some instances, the percentage of the common fluid buffer <NUM> in the ninth volume <NUM> is greater than a percentage of third fluid buffer <NUM> and/or reused third fluid buffer. During the third buffer reverse operation, the flow control valve <NUM> is set to connect the fluidic device <NUM> to the third fluid buffer reservoir <NUM> (e.g., by connecting valve selector channel <NUM> with the connecting channel <NUM> associated with the third fluid buffer reservoir <NUM>), and the pump <NUM> is operated to move fluid in a reverse direction through the fluidic device <NUM> to the third fluid buffer reservoir <NUM>. As shown in <FIG>, the ninth volume <NUM> of used fluid buffer is received back in the third fluid buffer reservoir <NUM> to be reused for one or more subsequent third wash operations. In some examples, the ninth volume <NUM> of used fluid buffer is less than or equal to the eighth volume <NUM> moved from the common fluid buffer reservoir <NUM>, in other examples, the ninth volume <NUM> of used fluid buffer is greater than the eighth volume <NUM> moved from the common fluid buffer reservoir <NUM>, and, in other examples, the ninth volume <NUM> of used fluid buffer is equal to the total amount of fluid buffer flushed through the fluidic device <NUM>, i.e., the combined seventh volume <NUM> moved from the third fluid buffer reservoir <NUM> and eighth volume <NUM> moved from the common fluid buffer reservoir <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a fourth reagent operation, such that flow control valve <NUM> permits fluid flow from the selected fourth reagent fluid reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>. During the fourth reagent operation, the flow control valve <NUM> is set to connect the fourth reagent fluid reservoir <NUM> to the fluidic device <NUM> (e.g., by connecting valve selector channel <NUM> with the corresponding connecting channel <NUM> associated with the fourth reagent fluid reservoir <NUM>). The pump <NUM> is operated to draw the fourth reagent fluid <NUM> from the fourth reagent fluid reservoir <NUM> through the flow control valve <NUM> and into the fluidic device <NUM>. As shown in <FIG>, an aliquot <NUM> of the fourth reagent fluid <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM>.

Referring to <FIG>, the system <NUM> may be set to perform a fourth reagent reverse operation, such that the pump <NUM> redirects at least a portion of the aliquot <NUM> of the fourth reagent fluid <NUM> moved in the fourth reagent operation back into the fourth reagent fluid reservoir <NUM>. During the fourth reagent reverse operation, the flow control valve <NUM> remains set to connect the fluidic device <NUM> to the fourth reagent fluid reservoir <NUM>, and the pump <NUM> is operated to expel fluid in a reverse direction through the fluidic device <NUM> to the fourth reagent fluid reservoir <NUM>. As shown in <FIG>, at least a portion <NUM> of the aliquot <NUM> of the fourth reagent fluid <NUM> moved in the fourth reagent operation is received back in the fourth reagent fluid reservoir <NUM> to be reused for subsequent fourth reagent operation.

In some examples, it may not be feasible or practical to reuse a fluid buffer after a wash operation - for example, if the characteristics of reagent or other material being flushed in the wash operation are such that the risk of carryover from re-using a buffer used to flush the reagent are too great. In one example, combining fluid buffer used to flush reagents in the fluidic device <NUM> after a scavenger process of the SBS operation with fresh buffer typically compromises the subsequent processes of the SBS operation, even if a trace amount of the used fluid buffer was added to the fresh buffer. In such a situation, buffer may be moved through the fluidic device, for example, from the common buffer reservoir <NUM>, but the used buffer is not thereafter reversed back to a sequestered fluid buffer reservoir. In some examples, the risk of reusing a fluid buffer is determined to be too great when the used fluid buffer exceeds a first contamination level, which may be determined by experimentation.

In some examples, the system <NUM> may reuse fluid buffer from the same designated reservoir in two different wash operations that follow two different reagent operations. For example, if the characteristics of the reagent or other material of a first reagent operation are benign, meaning that the presence of the reagent or the material would not affect or compromise another reagent operation (e.g., a second reagent operation), then the fluid buffer used to flush that reagent or material after the first reagent operation may be used again in the wash operation following the second reagent operation. In some examples, a used fluid buffer is determined be benign when the used fluid buffer is below a second contamination level, which may be determined by experimentation.

Referring to <FIG>, the system <NUM> may be set to perform a fourth wash operation, such that flow control valve <NUM> permits fluid flow from the selected common fluid buffer reservoir <NUM> of the set of reservoirs <NUM> to the fluidic device <NUM>. As shown in <FIG>, a volume <NUM> of the common fluid buffer <NUM> is moved through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or into a chamber of the pump <NUM> to flush the fourth reagent fluid <NUM> remaining in the fluidic device <NUM>. The system <NUM>, however, does not include a fluid buffer reservoir for storing and sequestering the buffer used in the fourth wash operation for possible re-use. Thus, the volume <NUM> of buffer moved through the fluidic device during the fourth wash operation is not reversed from the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>. Rather, the fluid buffer may be flushed through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM> and moved into a waste reservoir (not shown).

<FIG> illustrates a method <NUM> for sequestering and reusing reagent fluids and fluid buffers.

Method <NUM> comprises a step <NUM> of moving an aliquot of first reagent fluid <NUM> from the first reagent fluid reservoir <NUM> into the fluidic device <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the first reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first reagent fluid reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to move the first reagent fluid <NUM> from the first reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the first reagent fluid <NUM> through the fluidic device <NUM>.

Method <NUM> further comprises a first step <NUM> of moving a volume of unused common fluid buffer <NUM> into the fluidic device <NUM> where it is used. In some examples, step <NUM> includes a first part comprising moving a volume of the first fluid buffer <NUM> from the first dedicated fluid buffer reservoir <NUM> into the fluidic device <NUM> followed by a second part comprising moving a volume of the unused common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> into the fluidic device <NUM> where it is used. In some examples, the first part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the first fluid buffer <NUM> from the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>. In some examples, the first part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the first fluid buffer <NUM> through the fluidic device <NUM>.

In some examples, the second part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the common fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>. In some examples, the second part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the common fluid buffer <NUM> through the fluidic device <NUM>.

In some examples, the volume of first fluid buffer <NUM> moved from the first dedicated fluid buffer reservoir <NUM> is substantially equal to the volume of the common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>. In other instances, the volume of first fluid buffer <NUM> moved from the first dedicated fluid buffer reservoir <NUM> is greater than the volume of the common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>. In still other instances, the volume of first fluid buffer <NUM> moved from the first dedicated fluid buffer reservoir <NUM> is less than the volume of the common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>.

Method <NUM> further comprises a step <NUM> of moving at least a portion of the volume of the common fluid buffer <NUM> moved in step <NUM> and used in the fluidic device <NUM> back from the fluidic device <NUM> into the first dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit flow from the fluidic device <NUM> to the first dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to expel the fluid buffer from the fluidic device <NUM> to the first dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the second direction to generate a positive pressure differential to move the fluid buffer to the first dedicated fluid buffer reservoir <NUM>. In some examples, the portion of fluid buffer moved to the first dedicated fluid buffer reservoir <NUM> in step <NUM> ranges from about <NUM>% to <NUM>% of the total volume of fluid buffer moved into the fluidic device <NUM> in step <NUM>. For example, if <NUM>µL of first dedicated fluid buffer <NUM> is moved from the first dedicated fluid buffer reservoir <NUM> and <NUM>µL of common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>, then a volume of <NUM>µL to <NUM>µL of the combined <NUM>µL volume can be moved back into the first dedicated fluid buffer reservoir <NUM>. In some instances, the volume moved back into the first dedicated fluid buffer reservoir <NUM> can be predominantly common fluid buffer <NUM> based on the common fluid buffer121 displacing the first fluid buffer <NUM> further downstream through the fluidic system.

Method <NUM> further comprises a step <NUM> of moving an aliquot of second reagent fluid <NUM> from a second reagent fluid reservoir <NUM> into the fluidic device <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the second reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second reagent fluid reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to move the second reagent fluid <NUM> from the second reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the second reagent fluid <NUM> through the fluidic device <NUM>.

Method <NUM> further comprises a second step <NUM> of moving a volume of unused common fluid buffer <NUM> from the common fluid buffer source <NUM> into the fluidic device <NUM> where it is used. In some examples, step <NUM> includes a first part comprising moving a volume of the second fluid buffer <NUM> from the second dedicated fluid buffer reservoir <NUM> into the fluidic device <NUM> followed by a second part comprising moving a volume of the unused common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> into the fluidic device <NUM> where it is used. In some examples, the first part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the second fluid buffer <NUM> from the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second dedicated fluid buffer reservoir <NUM>. In some examples, the first part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the second fluid buffer <NUM> through the fluidic device <NUM>.

In some examples, the second part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the common fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to draw the volume of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>. In some examples, the second part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the common fluid buffer <NUM> through the fluidic device <NUM>.

Method <NUM> further comprises a step <NUM> of moving at least a portion of the volume of the fluid buffer moved in step <NUM> and used in the fluidic device <NUM> back from the fluidic device <NUM> into the second dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit flow from the fluidic device <NUM> to the second dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to expel the fluid buffer from the fluidic device <NUM> to the second dedicated fluid buffer reservoir <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the second direction to generate a positive pressure differential to move the fluid buffer to the second dedicated fluid buffer reservoir <NUM>. In some examples, the portion of fluid buffer moved to the second dedicated fluid buffer reservoir <NUM> in step <NUM> ranges from about <NUM>% to <NUM>% of the total volume of fluid buffer moved into the fluidic device <NUM> in step <NUM>. For example, if <NUM>µL of second fluid buffer <NUM> is moved from the second dedicated fluid buffer reservoir <NUM> and <NUM>µL of common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>, then a volume of <NUM>µL to <NUM>µL of the combined <NUM>µL volume can be moved back into the second dedicated fluid buffer reservoir <NUM>. In some instances, the volume moved back into the second dedicated fluid buffer reservoir <NUM> can be predominantly common fluid buffer <NUM> based on the common fluid buffer displacing the second fluid buffer <NUM> further downstream through the fluidic system.

Method <NUM> further comprises a step <NUM> of re-using the first reagent fluid <NUM> by moving an aliquot of the first reagent fluid <NUM> into the fluidic device <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the first reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first reagent fluid reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to move the first reagent fluid <NUM> from the first reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the first reagent fluid <NUM> through the fluidic device <NUM>.

Method <NUM> further comprises a step <NUM> of re-using the first fluid buffer by moving a volume of fluid buffer into the fluidic device <NUM>, where at least a portion of the volume of fluid buffer moved in step <NUM> is moved from the first fluid dedicated buffer reservoir <NUM>, which now includes used fluid buffer moved to first dedicated fluid buffer reservoir <NUM> in step <NUM>. In some examples, step <NUM> includes a first part comprising setting the flow control valve <NUM> to permit fluid flow from the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the first fluid buffer <NUM> from the first dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>. In some examples, the first part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the first fluid buffer <NUM> through the fluidic device <NUM>.

In some examples, step <NUM> includes a second part comprising, after moving the first fluid buffer <NUM>, moving a volume of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> into the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the common fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>. In some examples, the second part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the common fluid buffer <NUM> through the fluidic device <NUM>. In some examples, the volume of first fluid buffer <NUM> moved from the first dedicated fluid buffer reservoir <NUM> is substantially equal to the volume of the common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>.

In some examples, if the first fluid buffer <NUM> is to be further re-used after step <NUM>, the method comprises a step of moving at least a portion of the volume of the fluid buffer moved in step <NUM> back into the first dedicated fluid buffer reservoir <NUM>, as in step <NUM> above. In some examples, the step of moving at least a portion of the volume of the fluid buffer moved in step <NUM> back into the first dedicated fluid buffer reservoir <NUM> comprises setting the flow control valve <NUM> to permit flow from the fluidic device <NUM> to the first dedicated fluid buffer reservoir <NUM>. In some examples, the step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the first dedicated fluid buffer reservoir <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the first dedicated fluid buffer reservoir <NUM>. In some examples, the step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the first dedicated fluid buffer reservoir <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the second direction to generate a positive pressure differential to move the fluid buffer to the first dedicated fluid buffer reservoir <NUM>. In some examples, the portion of fluid buffer moved from the first dedicated fluid buffer reservoir <NUM> in step <NUM> ranges from about <NUM>% to <NUM>% of the volume of the fluid buffer moved into the fluidic device <NUM> in step <NUM>.

Method <NUM> may further comprise a step <NUM> of re-using the second reagent fluid <NUM> by moving an aliquot of the second reagent fluid <NUM> into the fluidic device <NUM>. In some examples, step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the second reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second reagent fluid reservoir <NUM>. In some examples, step <NUM> comprises using the pump <NUM> to draw move the second reagent fluid <NUM> from the second reagent fluid reservoir <NUM> to the fluidic device <NUM>. In some examples, step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the second reagent fluid <NUM> through the fluidic device <NUM>.

Method <NUM> may further comprise a step <NUM> of re-using the second fluid buffer by moving a volume of fluid buffer into the fluidic device <NUM>, where at least a portion of the volume of fluid buffer moved in step <NUM> is moved from the second dedicated fluid buffer reservoir <NUM>, which now includes used fluid buffer moved to second dedicated fluid buffer reservoir <NUM> in step <NUM>. In some examples, step <NUM> includes a first part comprising setting the flow control valve <NUM> to permit fluid flow from the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the second fluid buffer <NUM> from the second dedicated fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the first part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second dedicated fluid buffer reservoir <NUM>. In some examples, the first part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the second fluid buffer <NUM> through the fluidic device <NUM>.

In some examples, step <NUM> includes a second part comprising, after moving the second fluid buffer <NUM>, moving a volume of the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> into the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises setting the flow control valve <NUM> to permit fluid flow from the common fluid buffer reservoir <NUM> to the fluidic device <NUM> and using the pump <NUM> to move the common fluid buffer <NUM> from the common fluid buffer reservoir <NUM> to the fluidic device <NUM>. In some examples, the second part of step <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the common fluid buffer reservoir <NUM>. In some examples, the second part of step <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the first direction to generate a negative pressure differential to move the common fluid buffer <NUM> through the fluidic device <NUM>. In some examples, the volume of second fluid buffer <NUM> moved from the second dedicated fluid buffer reservoir <NUM> is substantially equal to the volume of the common fluid buffer <NUM> moved from the common fluid buffer reservoir <NUM>.

In some examples, if the second fluid buffer <NUM> is to be further re-used after step <NUM>, the method comprises a step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the second dedicated fluid buffer reservoir <NUM>, as in step <NUM>. In some examples, the step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the second dedicated fluid buffer reservoir <NUM> comprises setting the flow control valve <NUM> to permit flow from the fluidic device <NUM> to the second dedicated fluid buffer reservoir <NUM>. In some examples, the step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the second dedicated fluid buffer reservoir <NUM> comprises actuating the motor to rotate the flow control valve <NUM> to align and fluidly connect the valve selector channel <NUM> with a corresponding connecting channel <NUM> associated with the second dedicated fluid buffer reservoir <NUM>. In some examples, the step of moving at least a portion of the volume fluid buffer moved in step <NUM> back into the second dedicated fluid buffer reservoir <NUM> comprises using the actuator to move the plunger of the syringe pump <NUM> in the second direction to generate a positive pressure differential to move the fluid buffer to the second dedicated fluid buffer reservoir <NUM>.

As schematically shown in <FIG>, in some examples, the system <NUM> includes a fluid cartridge <NUM> supporting various components of the system <NUM>, such as, the fluidic device <NUM>, the flow control valve <NUM>, the pump <NUM>, and the set of fluid reservoirs <NUM>. The fluid cartridge <NUM> may be operatively installed into a processing instrument <NUM>. In some examples, the fluid cartridge <NUM> includes the inlet channel <NUM>, the flow control valve <NUM>, at least part of the outlet channel <NUM>, the set of fluid reservoirs <NUM>, and the set of connecting channels <NUM>. The fluidic device <NUM> may be operatively coupled to the instrument <NUM>, and the instrument <NUM> may include one or more actuators (e.g., motor) to control the position flow control valve <NUM> and the pressure applied by the pump <NUM> to select and move various reagent fluids and fluid buffers. Instrument <NUM> may further include a waste outlet (not shown) to dispose any used reagent fluid or fluid buffer. Controller <NUM>, which may be part of the instrument <NUM> or may be a standalone or remote computer resource operatively connected to the instrument <NUM>, controls operation of the instrument <NUM> (e.g., processing of the fluidic device <NUM> and operation of the pump <NUM>) and operation of the fluid cartridge <NUM> (e.g., operation of the flow control valve <NUM>).

Aspects of the disclosure are implemented via control and computing hardware components, user-created 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 effect computational and/or control steps by receiving one or more input values, executing one or more algorithms, as the algorithm described in <FIG>, stored on non-transitory machine-readable media (e.g., software) that provide instruction for manipulating or otherwise acting on the input values, and output one or more output values. Such outputs may be displayed or otherwise indicated to a user for providing information to the user, for example information as to the status of the instrument or a process being performed thereby, or such outputs may comprise inputs to other processes and/or control algorithms. Data input components comprise elements by which data is input for use by the control and computing hardware components. Such data inputs may comprise positions sensors, motor encoders, as well as manual input elements, such as graphic user interfaces, keyboards, touch screens, microphones, switches, manually-operated scanners, voice-activated input, etc. Data output components may comprise hard drives or other storage media, graphic user interfaces, monitors, printers, indicator lights, or audible signal elements (e.g., buzzer, horn, bell, etc.). Software comprises instructions stored on non-transitory computer-readable media which, when executed by the control and computing hardware, cause the control and computing hardware to perform one or more automated or semi-automated processes.

The system includes a control system including a computer controller <NUM> (schematically represented in <FIG>). Controller <NUM> may be a control system or computer connected to any one of the devices of the system <NUM>, e.g., a stand-alone computer, or may include computer components integrated with any one of the devices of the system <NUM>, e.g., an application specific integrated circuit. These computer components can include one or more microprocessors, displays, keyboards (and/or other user input devices), memory components, printer(s), and/or other devices. Controller <NUM> may be configured to receive inputs from a user (e.g., user-inputs) and/or feedback devices, such as pressure sensors, flow meters, etc., and manage the performance of the fluid operations of the system <NUM>. Controller <NUM> may include software algorithms to implement processes, such as a process implementing the method <NUM> shown in <FIG>, that enable a user to enter user-defined parameters related to fluid processing operations into the fluidic device <NUM> of the system <NUM>, schedule different fluid processing operations on the fluidic device <NUM> of the system <NUM>, and/or cause the controller <NUM> to perform the different steps associated with the fluid processing operations, monitor the performance of the fluid processing operations, and/or output results (on display, printout, etc.) for the user.

As shown in <FIG>, the controller <NUM> is in electrical communication with the flow control valve <NUM>, the pump <NUM> (indicated by the dashed lines), and/or intermediary devices configured to control the flow control valve <NUM> and/or the pump <NUM> (e.g., a step motor for the flow control valve <NUM>, a motor for the pump <NUM>, etc.) such that the controller <NUM> may send instructions to control the control valve <NUM> and the pump <NUM> to perform different steps associated with the fluid processing operations (e.g., the processes associated with <FIG> and the method of <FIG>). The controller <NUM> is configured to transmit a command for the flow control valve <NUM> to fluidly connect a selected fluid reservoir of the set of reservoirs <NUM> to the inlet channel <NUM> so that the fluid from the selected fluid reservoir may flow through the corresponding connecting channel <NUM>, the flow control valve <NUM>, the inlet channel <NUM>, the fluidic device <NUM>, the outlet channel <NUM>, and/or a chamber of the pump <NUM>. In some examples, the controller <NUM> is configured to transmit a command to the pump <NUM> to move in the first or second direction to generate a pressure differential between the any one of the set of the fluid reservoirs <NUM> and the outlet channel <NUM> to drive fluid flow towards or away from the pump <NUM>.

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

Claim 1:
A method comprising:
(a) moving (<NUM>) an aliquot of first reagent fluid (<NUM>) from a first reagent fluid reservoir (<NUM>) into a fluidic device (<NUM>);
(b) after step (a), moving (<NUM>) a volume of unused common fluid buffer (<NUM>) from a common fluid buffer source (<NUM>) into the fluidic device (<NUM>) where it is used;
(c) after step (b), moving (<NUM>) at least a portion of the volume of common fluid buffer (<NUM>) moved in step (b) and used in the fluidic device (<NUM>) back from the fluidic device (<NUM>) into a first dedicated fluid buffer reservoir (<NUM>);
(d) after step (c), moving (<NUM>) an aliquot of second reagent fluid (<NUM>) from a second reagent fluid reservoir (<NUM>) into the fluidic device (<NUM>);
(e) after step (d), moving (<NUM>) a volume of unused fluid buffer from the common fluid buffer source (<NUM>) into the fluidic device (<NUM>) where it is used; and
(f) after step (e), moving (<NUM>) at least a portion of the volume of fluid buffer moved in step (e) and used in the fluidic device (<NUM>), back from the fluidic device (<NUM>) into a second dedicated fluid buffer reservoir (<NUM>);
(g) after step (f), moving (<NUM>) an aliquot of the first reagent fluid (<NUM>) into the fluidic device (<NUM>);
(h) after step (g), moving (<NUM>) a volume of fluid buffer into the fluidic device (<NUM>), wherein at least a portion of the volume of fluid buffer is moved from the first dedicated fluid buffer reservoir (<NUM>) and has been used in the fluidic device (<NUM>).