阅读说明：本技术 用于流体递送以进行测序的装置、系统和方法 (Apparatus, system, and method for fluid delivery for sequencing ) 是由 A·卡里洛 J·多诺赫 于 2020-05-29 设计创作，主要内容包括：本发明教导的流体系统的各个实施例被配置成在下一代测序分析的过程中执行一系列流体操作,以将在分析过程中使用的各种溶液依序递送到多泳道传感器装置。示例性流体操作包含洗涤、灌注和通过流体多路复用器块进行核苷酸试剂递送,所述流体多路复用器块被配置成向用于在分析期间进行检测的多泳道传感器装置的每个泳道提供独立的流体分配。因此,在分析期间可以使用任何数量的泳道或泳道的任何组合,使得在分析期间,可以在运行期间单独地使用任何位置中的一个泳道,可以在运行期间同时使用所有四个泳道,或者可以在运行期间同时使用泳道的任何组合。(Various embodiments of the fluidic system taught by the present invention are configured to perform a series of fluidic operations during a next generation sequencing analysis to sequentially deliver various solutions used in the analysis process to a multi-lane sensor device. Exemplary fluidic operations include washing, perfusion, and nucleotide reagent delivery through a fluidic multiplexer block configured to provide independent fluid assignments to each lane of a multi-lane sensor device for detection during analysis. Thus, any number of lanes or any combination of lanes may be used during the analysis, such that during the analysis, one lane in any position may be used individually during the run, all four lanes may be used simultaneously during the run, or any combination of lanes may be used simultaneously during the run.)

1. A fluidic device, comprising:

a fluidic multiplexer block comprising a plurality of fluidic multiplexer units, each fluidic multiplexer unit comprising a substrate having a fluidic distribution circuit formed therein, the fluidic distribution circuit comprising a plurality of fluidic branches;

a first fluid interface block assembly connected to a first face of the fluid multiplexer block, the first fluid interface block including a first set of flexible tubes and a second set of flexible tubes, each set of tubes connected to each corresponding port of a first set of ports and a second set of ports on the first face of the fluid multiplexer block, respectively;

a second fluid interface block assembly connected to a second face of the fluid multiplexer block opposite the first face, the second fluid interface block including a first set of flexible tubes and a second set of flexible tubes, each set of tubes connected to each corresponding port of a first set of ports and a second set of ports on the second face of the fluid multiplexer block, respectively;

a third fluid interface block assembly connected to a third face of the fluid multiplexer block adjoining the first and second faces, the third fluid interface block including a first set of flexible tubes and a second set of flexible tubes, each set of tubes connected to each corresponding port of a first set of ports and a second set of ports, respectively, on the third face of the fluid multiplexer block; and

a fourth fluid interface block assembly connected to a third face adjacent to the third interface block assembly, the fourth fluid interface block including a first set of flexible tubing and a second set of flexible tubing, each set of tubing connected to a port of the fluid multiplexer block corresponding to each of a third set of ports and a fourth set of ports on the third face.

2. The fluidic device of claim 1, wherein the device is operably connected to a sequencing system.

3. The fluidic device of claim 2, wherein said first set of flexible tubing of said first fluid interface block assembly provides fluid communication between each first fluid branch of each fluid multiplexer unit's first fluid branch and a first nucleotide reagent source, and said second set of flexible tubing of said first fluid interface block assembly provides fluid communication between each second fluid branch of each fluid multiplexer unit's second fluid branch and a second nucleotide reagent source.

4. The fluidic device of claim 2, wherein said first set of flexible tubes of said second first fluid interface block assembly provide fluid communication between each of the third fluid legs of each fluid multiplexer unit and a third nucleotide reagent source, and said second set of flexible tubes of said second fluid interface block assembly provide fluid communication between each of the fourth fluid legs of each fluid multiplexer unit and a fourth nucleotide reagent source.

5. The fluidic device of claim 2, wherein the first set of flexible tubes of the third fluidic interface block assembly provide fluid communication between each of the fifth fluidic branches of each fluidic multiplexer unit and a source of calibration solution, and the second set of flexible tubes of the third fluidic interface block assembly provide fluid communication between each of the primary waste channels of each fluidic multiplexer unit and a waste container.

6. The fluidic device of claim 2, wherein said first set of flexible tubing of said fourth fluidic interface block assembly provides fluid communication between each of the wash solution channels of each fluid multiplexer unit and a wash solution source, and said second set of flexible tubing of said fourth fluidic interface block assembly provides fluid communication between each of the main sensor device waste channels of each fluid multiplexer unit and a waste container.

7. The fluidic device of claim 2, further comprising a set of sensor device interface connector ports located on a fourth face of the fluidic multiplexer block.

8. The fluidic device of claim 7, wherein the set of sensor interface connector ports of the fourth face of the fluidic multiplexer block includes a set of sensor device interface inlet connector ports and a set of sensor device interface outlet connector ports.

9. The fluidic device of claim 8, wherein the fluidic device is operably connected to a multi-lane sensor device such that:

a set of sensor device inlet ports of the multi-lane sensor device are connected and sealed to each respective one of the set of sensor device interface inlet connector ports of the fourth face of the fluid multiplexer block; and is

A set of sensor device outlet ports of the multi-lane sensor device are connected and sealed to each respective one of the set of sensor device interface outlet connector ports of the fourth face of the fluid multiplexer block.

10. The fluidic device of claim 1, wherein the substrate is a polymeric material selected from the group consisting of polycarbonate, polymethylmethacrylate, polyetherimide, and polyimide.

11. A fluidic control system for a sequencing system, the fluidic control system comprising:

a plurality of input source vessels and at least one output vessel, wherein the plurality of input source vessels contain a solution for a sequencing run;

a fluid multiplexer block assembly located in a fluid path between a fluid distribution manifold assembly and the at least one output container, wherein the fluid distribution manifold controls the distribution of the solution for a sequencing run; and

a fluid regulation and control system, wherein the fluid regulation and control system provides a defined and controllable pressure differential between the plurality of input source vessels and the output vessel.

12. The fluid control system of claim 11, further comprising:

a fluid handling manifold, wherein the fluid handling manifold controls the distribution of solutions for cleaning and filling operations.

13. The fluid control system of claim 12, further comprising:

a reagent concentrate cartridge for preparing a bulk reagent in a bulk reagent container assembly, wherein the reagent concentrate cartridge is located in a fluid path between the fluid handling manifold and the bulk reagent container assembly.

14. The fluid control system of claim 11, wherein the fluid multiplexer block assembly is configured to reversibly couple to a sensor device.

15. A method for automated fluidic system workflow for a sequencing system, the method comprising:

preparing bulk nucleotides and calibration solutions from reagent concentrate cartridges;

priming each of a plurality of fluid pathways of a fluid system;

calibrating the sensor device with a wash solution and a calibration solution;

controlling the flow of the nucleotide solution and the wash solution through the sensor device during a sequencing run; and

the fluidic system is cleaned between sequencing runs.

16. The method of claim 15, further comprising repeating a perfusion bath cleaning for subsequent runs until the multi-lane sensor device is used or otherwise depleted.

17. The method of claim 16, further comprising cleaning the fluid system using a used or otherwise depleted sensor device.

SUMMARY

Next Generation Sequencing (NGS) is a high throughput method that enables rapid sequencing of base pairs in DNA or RNA samples. The scale and efficiency of NGS affects a wide range of applications including gene expression profiling, chromosome counting, epigenetic change detection, and molecular analysis. As a result, performance of NGS is being used by researchers in many disciplines to accelerate the future of finding pace and achieving personalized medicine.

To accommodate the ever-increasing pace of discovery, the demand for increased size and throughput of NGS systems is also increasing. This increase in scale and throughput is accompanied by a need for sample input-answer output solutions, making automated quality control and scale a consideration for NGS systems. Accordingly, there is a need in the art for NGS fluid systems and related components to meet demand.

Drawings

A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth hereinafter and by reference to the accompanying drawings, in which:

fig. 1 is an exploded view generally illustrating a fluidic multiplexer block.

Fig. 2 is an isometric view of a fluid-multiplexer cell generally illustrating a fluid-multiplexer block, such as the fluid-multiplexer block of fig. 1.

Figure 3A is a perspective view illustrating generally a multi-lane sensor apparatus.

Figure 3B is a schematic representation showing generally the fluidic integration between the fluid multiplexer unit and a selected lane of the multi-lane sensor device.

Fig. 4 is an isometric view generally illustrating the integration of a fluidic interface block with a fluidic multiplexer block, such as the fluidic multiplexer block of fig. 1.

Fig. 5A is an isometric view generally illustrating a fluid interface block.

Fig. 5B is a cross-sectional view generally illustrating the seal formed between the fluid lines of the fluid interface block and the ports of the fluid multiplexer unit.

FIG. 6 is a schematic representation of a fluidic system generally illustrating a sequencing system.

Fig. 7 is a rear isometric view generally showing a fluid multiplexer block clamp assembly including a fluid multiplexer block clamp with a fluid multiplexer block assembly mounted therein.

Fig. 8A is a cross-sectional view generally illustrating the integration of electrodes into a fluid multiplexer unit. Fig. 8B is an expanded view of the electrode of fig. 8A.

Fig. 9 is a front isometric view generally showing a fluid multiplexer block clamp assembly including a fluid multiplexer block clamp with a fluid multiplexer block assembly mounted therein.

FIG. 10 is a cross-sectional view of an assembly generally showing a fluid multiplexer block mounted into a fluid multiplexer block fixture of a multi-lane sensor device positioned in a sensor device mounting assembly.

Figure 11 is an expanded isometric view generally illustrating the mounting of a fluid multiplexer block to a multi-lane sensor device.

FIG. 12 is a block diagram generally illustrating a sequencing system.

FIG. 13 is a perspective view illustrating a sequencing system generally.

FIG. 14 is a perspective view of a container cabinet generally illustrating a sequencing system.

Fig. 15 generally illustrates a flow diagram of a method for automated fluid delivery in a sequencing system.

Detailed Description

Fig. 1 generally illustrates an exploded view of a fluidic multiplexer block 100 that, as a component of the fluidic system of an integrated next generation sequencing system, can provide for the distribution of various solutions used during analysis to a multi-lane sensor device. Various embodiments of the fluidic systems disclosed herein are configured to perform a series of fluidic operations to sequentially deliver various solutions to a multi-lane sensor device during a next generation sequencing analysis. Exemplary fluidic operations include washing, priming, and nucleotide reagent delivery through a fluidic multiplexer block, such as fluidic multiplexer block 100 of fig. 1. Such a fluid multiplexer block is configured to provide an independent fluid distribution to each lane of a multi-lane sensor device for detection during analysis. Any number of lanes or any combination of lanes may be used during the analysis in accordance with the teachings of the present invention, such that during the analysis, one lane in any position may be used individually during the run, all lanes may be used simultaneously during the run, or any combination of lanes may be used simultaneously during the run. Using the fluid multiplexer block taught by the present invention for distributing fluids to a multi-lane sensor device during a series of fluidic operations may avoid cross-contamination of the solutions used during analysis in the individual fluidic compartments as well as provide sharp transitions between reagent fluid flows during analysis. Additionally, various embodiments of the fluid multiplexer block may provide a constant electrolyte fluid environment for the reference electrode, thereby providing a constant stable reference voltage for the multi-lane sensor device.

As depicted in fig. 1, the fluid-multiplexer block 100 includes fluid-multiplexer cells 200A-200D and first and second end caps 105A, 105B. In accordance with the teachings of the present invention, each fluid multiplexer unit has a fluid multiplexer circuit formed within the body of the fluid multiplexer unit. Thus, as depicted in fig. 1, each of fluid multiplexer cells 200A-200D has a fluid multiplexer circuit 215A-215D formed within the body of each fluid multiplexer cell. As will be provided in more detail later herein, each fluid multiplexer unit in the fluid multiplexer block 100 is independently controllably in fluid communication with one of each of the flow cell lanes of the multi-lane sensor device. Thus, a first lane of the multi-lane sensor device may be fluidically integrated into fluid multiplexer unit 200A, while a second lane may be fluidically integrated into fluid multiplexer unit 200B, and a third lane may be fluidically integrated into fluid multiplexer unit 200C, while a fourth lane may be fluidically integrated into fluid multiplexer unit 200D. Further, any number of lanes or any combination of lanes may be used during the analysis, such that during the analysis setup, the end user may select one lane in any position used individually during the run, all lanes used simultaneously during the run in the order specified by the end user, or any combination of lanes used simultaneously during the run in the order specified by the end user.

Fig. 2 generally illustrates an embodiment of a fluid multiplexer unit 200 that contains four input reagents and a calibration solution in each of five fluidic branches and has a dispensing channel for a wash solution. The fluidic circuit 215 is formed in a substrate 205 having a first surface 201 and an opposing second surface 203. As depicted in fig. 2, the first surface 201 and the opposing second surface 203 are substantially parallel to each other. Fluid multiplexer unit 200 may have a first fluid interface side 202 and an opposite second fluid interface side 204. As depicted in fig. 2, a third fluid interface side 206 connects the first and second interface edges on one side, and a fourth fluid interface side 208 connects the first and second interface edges on an opposite side of the third fluid interface side 206. The substrate 205 may be constructed of a variety of materials such as glass, ceramic, and plastic. Exemplary polymeric materials include polycarbonate, polymethyl methacrylate, polyetherimide and polyimide. Reagent inlet ports 210, 216, 222, and 228 and calibration solution inlet port 234 are in fluid communication with inlet channels 211, 217, 223, 229, and 235, respectively. The inlet channels 211, 217, 223, 229 are in fluid communication with the curvilinear channels 213, 219, 225, 231, and 235, respectively, of each of the five fluid branches. Finally, the wash solution inlet port 240 is in fluid communication with the wash solution channel 242.

As depicted in fig. 2, each inlet channel forms a three-way junction with each curvilinear channel, such that each curvilinear channel consists of two branches. Such a three-way joint forming two branches is depicted in fig. 2, wherein the inlet channel 211 is three-way into a curved channel 213, forming a first branch channel 212 and a second branch channel 214. Similarly, the inlet passage 217 is ported into a curved passage 219, forming a first bypass passage 218 and a second bypass passage 220, while the inlet passage 223 is ported into a curved passage 225, forming a first bypass passage 224 and a second bypass passage 226. Additionally, the inlet channel 229 is three-way into the curved channel 231, thereby forming a first branch channel 230 and a second branch channel 232. Finally, the inlet channel 235 leads into a curved channel 237, thereby forming a first bypass channel 236 and a second bypass channel 238. First branch channels 212, 218, 224, 230, and 236 of the curvilinear channels 213, 219, 225, 231, and 237 of the five fluid branches, respectively, are in fluid communication with the central channel 250. As depicted in fig. 2, central channel 250 is in fluid communication with a sensor device interface inlet connector port 260, which is in fluid communication with a sensor inlet port (not shown). Additionally, the wash solution inlet port 240 is in fluid communication with a wash solution channel 242, which is also in fluid communication with the sensor device interface inlet connector port 260. Sensor device interface outlet connector port 262 is in fluid communication with a sensor outlet port (not shown) and sensor device waste passage 244. The sensor unit waste channel 244 is in fluid communication with a sensor unit waste receptacle (not shown) that is connected to the fluid multiplexer unit 200 through a sensor unit waste port outlet 264. Each of the second bypass channels 214, 220, 226, 232, and 238 of curvilinear channels 213, 219, 225, 231, and 237, respectively, is in fluid communication with a main waste channel 246, which is in fluid communication with a main waste receptacle (not shown) connected to fluid multiplexer unit 200 through a main waste outlet port 266.

FIG. 3A is a perspective view illustrating generally a multi-lane sensor device, such as a microarray device comprising a flow cell. For example, as depicted in fig. 3A, the sensor device 10 includes a die 4 mounted on a substrate 2. The die 4 may have a plurality of micro-holes in fluid communication with the sensor array. As used herein, an "array" is a planar arrangement of elements such as sensors or microwells. The array may be one-dimensional or two-dimensional. A one-dimensional array may be an array having a column (or row) of elements in a first dimension and a plurality of column (or row) elements in a second dimension. The number of columns (or rows) in the first dimension and the second dimension may be the same or different. Further, embodiments of the sensor device may include a plurality of micro-holes disposed over a Field Effect Transistor (FET) sensor array. In various embodiments of the sensor device, the microwell may be a reaction chamber that provides a containment or confinement region for sequencing. In this aspect, the microwell array may comprise a plurality of microwells loaded with one or more polymer particles or beads prepared for sequencing a target polynucleotide sample, each loaded microwell being disposed over at least one FET sensor. In various embodiments of the sensor device, the FET sensor may be a chemical sensitive FET (chemfet). For various embodiments of the sensor device, the FET sensor may be an ion sensitive FET (isfet). Both chemFET sensors and ISFET sensors can have structural analogs of MOSFET transistors in which the charge on the gate electrode is generated by chemical processes such as incorporation of nucleotides during sequencing by synthesis. In this regard, ISFETs can be used to measure protons (i.e., pH) released in microwells due to incorporation of nucleotides during a sequencing reaction. A sensor device as taught by the present invention, such as sensor device 10 of FIG. 3A, may comprise 10²1, 10³1, 10⁴1, 10⁵1, 10⁶1, 10⁷An array of one or more FET sensors.

The flow cell 6 of fig. 3A is mounted securely over the substrate 2, providing a volume over the die 4. In an example, flow cell 6 includes a set of fluid inlets, such as inlets 3A-3D, and a set of fluid outlets, such as outlets 5A-5D, as depicted in fig. 3A. The flow cell 6 may be divided into lanes, such as lanes 4A-4D, where each lane is individually in fluid communication with a respective fluid inlet and fluid outlet. For example, as depicted in fig. 3A, lane 4A is in fluid communication with inlet 3A and outlet 5A. As shown, the sensor device 10 includes four lanes. Alternatively, the sensor device 10 may comprise less than four lanes or more than four lanes. For example, the sensor device 10 may comprise between 1 and 10 lanes, such as between 2 and 8 lanes, or 4 to 6 lanes. For sensor devices taught by the present invention, the lanes of a multi-lane sensor device may be fluidically isolated from each other. Thus, lanes such as lanes 4A-4C of FIG. 3A may be used at separate times or simultaneously, depending on aspects of the end user defined sequencing run plan. As will be provided in more detail later herein, the sensor device 10 may further include a guide structure, such as, for example, alignment pins 12A and 12B formed as part of the flow cell 1406. Alignment pins 12A and 12B may engage complementary structures on the fluid multiplexer block. Such alignment pins may help align fluid inlets and fluid outlets, such as 3A-3D and 5A-5D of fig. 3A, respectively, with associated ports on a fluid multiplexer block, such as fluid multiplexer block 100 of fig. 1. In accordance with the teachings of the present invention, an automated fluidic system can provide a sequence of nucleotide reagents flowing through a flow cell, such as flow cell 6 of fig. 3A, where each reagent contains a single type of nucleotide. As previously described herein, in response to nucleotide addition during the sequencing reaction, the pH within the local environment of the microwell may change and may be detected by the ISFET sensors in the sensor array. Thus, a change in pH associated with a known sequence of nucleotide reagent flow can be used to indicate the sequence of nucleotides complementary to a polynucleotide sample on a particle or bead that has been prepared for sequencing a polynucleotide sample of interest.

Figure 3B generally shows a schematic representation of the fluidic integration of the fluidic multiplexer unit 200 of the fluidic multiplexer block 100 of figure 1 with a multi-lane sensor arrangement.

With respect to fluid delivery and control for performing various analyses on a multi-lane sensor device, such as the sensor device 10 of fig. 3B, the fluid circuit 215 of the fluid multiplexer unit 200 may be in fluid communication with one flow cell lane of the sensor device 10. For illustrative purposes, one fluid multiplexer unit is shown in fig. 3B fluidly integrated with one flow cell lane. However, the end user may select any number of lanes or any combination of lanes during the analysis, as each lane is fluidically integrated with a fluid multiplexer unit, such as one of fluid multiplexer units 200A-200D of fig. 1. As a non-limiting example, a first flow cell lane, such as flow cell lane 4A of sensor device 10 of fig. 3B, may be fluidically integrated with a first fluid multiplexer, such as fluid multiplexer unit 200A of fig. 1, while a second flow cell lane, such as flow cell lane 4B of sensor device 10 of fig. 3B, may be fluidically integrated with a second fluid multiplexer, such as fluid multiplexer unit 200B of fig. 1. Similarly, a third flow cell lane, such as flow cell lane 4C of sensor device 10 of fig. 3B, may be fluidically integrated with a third fluid multiplexer, such as fluid multiplexer unit 200C of fig. 1, while a fourth flow cell lane, such as flow cell lane 4D of sensor device 10 of fig. 3B, may be fluidically integrated with a fourth fluid multiplexer, such as fluid multiplexer unit 200D of fig. 1. In this regard, for the purposes of illustration of fig. 3B, the description herein generally relates to how each fluid multiplexer unit 200 is fluidically integrated with each of the corresponding flow cell lanes of the multi-lane sensor device.

As will be provided in more detail later herein, the fluidic system of the sequencer may include a plurality of solution containers that provide a plurality of solutions for use during a sequencing run. For example, the various solutions may contain various nucleotide reagents for analysis, calibration solutions, diluent (wash) solutions, and cleaning solutions. Various solutions used during a sequencing run may be passed through a flow cell inlet, such as flow cell inlet 3A of flow cell lane 4A of FIG. 3B, and any of sensor devices 10Which flow cell lanes are controllably fluidly connected. The fluidic system of a sequencer in accordance with the teachings of the present invention may include a fluid line from each of the solution containers that may be selectively placed in fluid communication with an inlet channel of fluidic circuit 215, such as inlet channels 211, 217, 223, 229, 235, and 242. In addition, as depicted in fig. 3B, the fluid flow of each of the various solutions used in the analysis process may be controlled by a valve, such as a separate fluid line L for the reagents₁To L₄Each reagent fluid line of (a) and a line for calibration solution L₅And a washing solution line L₆Fluid line valve V₁To V₆To control. It should be noted that the fluid line valve V is calibrated except during a calibration sequence prior to run start-up to calibrate the sensor device selected for use during operation₅Typically in the closed position. Thus, calibrating the fluid line valve V₅Is off during the sequencing run.

In conjunction with the controllable flow of various solutions used in the analysis process, fluid multiplexer unit 200 of fig. 3B may perform fluidic operations including, for example and without limitation, providing selected reagent delivery to a flow cell lane of sensor device 10, washing fluid multiplexer circuit 215 and a flow cell, such as flow cell lane 4A of sensor device 10, and priming fluid multiplexer circuit 215 with the selected reagent. Such fluidic manipulation may provide for cross-contamination free delivery of reagents to a flow cell, such as flow cell lane 4A of fig. 3B, may provide for sharp transitions between reagent fluid flows, and provide a constant electrolyte fluid environment for the reference electrode 275 shown in fig. 3B in fluid communication with the central channel 250, thereby providing a constant stable reference voltage to the sensor device 10.

For example, the fluid multiplexer unit 200 of fig. 3B may selectively provide reagent fluid lines L₁To L₄Is in fluid communication with the first flow cell inlet 3A of the first flow cell lane 4A, thereby providing selective reagent flow through the first flow cell lane 4A of the sensor device 10. Is notThe restrictive illustrative reagent fluid path is given by a reagent delivery operation in which a wash solution fluid line valve V₆In a closed state and a reagent fluid line valve V₁To V₄One of which is in an open state, provided that one of the selected reagents is in fluid communication with fluid multiplexer circuit 215. Under such conditions, the selected reagent may flow through fluid multiplexer circuit 215 and then be discarded through waste channel 246. Additionally, the selected reagent may flow through the fluid multiplexer circuit 215 to the first flow cell inlet 3A of the first flow cell lane 4A, where it may then flow through the first flow cell lane 4A to the first outlet port 5A, and finally through the flow cell outlet line (see fig. 2) to the flow cell waste container.

With respect to fluid control of the wash solution, the fluid multiplexer unit 200 of fig. 3B may selectively provide fluid communication between the wash solution and the first flow cell lane 4A. Thus, in the wash solution fluid line valve V₆In the open state, the washing solution line L₆Can be in fluid communication with fluid multiplexer waste channel 246 and flow cell waste channel (see fig. 1) provided that a wash of fluid multiplexer circuit 215 and first flow cell lane 4A can be performed. A non-limiting illustrative wash solution fluid path is given by a wash operation in which a wash solution fluid line valve V₆In an open state and a reagent fluid line valve V₁To V₄Provided that the wash solution can flow through the wash solution fluid passage 242 to the three-way junction having the central passage 250. Since the central channel 250 is in fluid communication with the waste channel 246 through the fluid multiplexer circuit 215, the wash solution can flow through the waste channel 246 to the fluid multiplexer waste. As will be provided in more detail later herein, the wash solution may pass through the first flow cell lane 4A from the first inlet port 3A to the first outlet port 5A, and then to the flow cell waste container.

Priming fluid multiplexer circuit 215 of fluid multiplexer unit 200 with the selected reagent may be performed, for example, sequentially after a wash operation and before the selected reagent is placed in fluid communication with first flow cell lane 4A of fig. 3B. A non-limiting example illustrating reagent priming is given by a reagent priming operation in which the solution fluid line valve V₆In an open state and a reagent fluid line valve V₁To V₄One of the selected reagents is selected to be in an open state, provided that the selected one of the reagents is in fluid communication with fluid multiplexer circuit 215 of fluid multiplexer unit 200. In this operation, the flow rate of the wash solution is selected relative to the flow rate of the reagents such that the wash solution flows through the wash channel 242 and through the first flow cell lane 4A of the sensor device 10 to the sensor device waste. Under such conditions, the selected reagent circulates through the fluid multiplexer circuit 215 as it is prevented from flowing through the sensor device 10 by the wash solution flowing through the device. Thus, the selected reagent flows through fluid multiplexer circuit 215 to the main waste through waste channel 246. Thus, when an agent delivery operation as previously provided herein is initiated, the agent selected in the agent perfusion operation is in direct flow communication with first flow cell inlet 3A.

Accordingly, various embodiments of the fluidic system are configured to perform a series of operations to sequentially deliver various solutions to the sensor device during a next generation sequencing analysis. For example, a series of operations may include washing, priming, and delivering a nucleotide reagent to the sensor device through the fluid multiplexer block unit, as depicted in fig. 3B. Using a fluidic multiplexer block for dispensing fluids to a sensor device during a series of fluidic operations as taught by the present invention can avoid cross-contamination of reagents in individual fluidic compartments as well as provide sharp transitions between reagent fluid flows. Additionally, various embodiments of the fluidic system may provide a constant electrolyte fluid environment for the reference electrode, thereby providing a constant stable reference voltage to the sensor device.

Fig. 4 generally shows a fluidic multiplexer block assembly 110 that includes a fluidic multiplexer block 100 and each of fluidic interface block assemblies 300, 310, 320, and 330. Each interface block assembly may include a fluid interface block, such as fluid interface blocks 302, 312, 322, and 332 of fluid interface block assemblies 300, 310, 320, and 330, respectively. Each fluid interface block has first and second sets of flexible tubes connected to the block, such as first and second flexible tube sets 304, 306 connected to the fluid interface block 302, first and second flexible tube sets 314, 316 connected to the fluid interface block 312, first and second flexible tube sets 324, 326 connected to the fluid interface block 322, and first and second flexible tube sets 334, 336 connected to the fluid interface block 332.

As depicted in fig. 4, each fluid interface block is mounted to a face of the fluid multiplexer block 100 such that each fluid line in each set of flexible tubes can be coupled and sealed to a respective inlet port in a complementary set of ports. For example, in fig. 4, the fluid interface block assembly 300 is shown mounted on a first face 102 of the fluid multiplexer block 100, while the fluid interface block assembly 310 is shown mounted on an opposing second face 104. Similarly, fluid interface block assemblies 320 and 330 are shown mounted on third face 106, which abuts first face 103 and second face 104 of fluid-multiplexer block 100. For each fluid interface block, each fluid line in each set of flexible tubes may be coupled and sealed to a respective inlet port in a complementary set of ports on each face of the fluid multiplexer block 100. For example, as shown in the exploded view of fig. 1, each fluid multiplexer unit 200A-200B provides a first set of ports and a second set of ports for an assembled fluid multiplexer block.

In this regard, a fluid interface block assembly 300 mounted on the fluid multiplexer block first face 102 has a first flexible tube set 304 and a second flexible tube set 306. For each fluid multiplexer block unit of the fluid multiplexer block 100, each tube of the flexible tube set 304 is connected to a corresponding first reagent inlet port, such as the first nucleotide reagent inlet port 210 of the fluid multiplexer unit 200 of fig. 2, and each tube of the flexible tube set 306 is connected to a corresponding second reagent inlet port, such as the second nucleotide inlet port reagent 216 of the fluid multiplexer unit 200 of fig. 2. As depicted in fig. 4, a fluid interface block assembly 310 mounted on the fluid multiplexer block second face 104 has a first flexible tube set 314 and a second flexible tube set 316. For each fluid multiplexer block unit of the fluid multiplexer block 100, each tube of the flexible tube set 314 is connected to a corresponding third reagent inlet port, such as the third nucleotide reagent inlet port 222 of the fluid multiplexer unit 200 of fig. 2, and each tube of the flexible tube set 316 is connected to a corresponding fourth reagent inlet port, such as the fourth nucleotide reagent inlet port 228 of the fluid multiplexer unit 200 of fig. 2. Thus, each tube of the flexible tube set 304 may be coupled and sealed to a first set of ports on the first face 102, while each tube of the flexible tube set 306 may be coupled and sealed to a second set of ports on the first face 102. In a corresponding manner, each tube of the flexible tube set 314 may be coupled and sealed to a first set of ports on the second face 104, while each tube of the flexible tube set 316 may be coupled and sealed to a second set of ports on the second face 104.

Similarly, fig. 4 depicts a fluid interface block assembly 320 mounted on a fluid multiplexer block third face 106 that is orthogonal to and abuts first face 102 and second face 104. The fluid interface block assembly 320 has a first flexible tube set 324 and a second flexible tube set 326. For each fluid multiplexer block unit of the fluid multiplexer block 100, each tube of the flexible tube set 324 is connected to a corresponding sensor device waste outlet port, such as sensor device waste outlet port 264 of the fluid multiplexer unit 200 of fig. 2, and each tube of the flexible tube set 326 is connected to a corresponding wash solution inlet port, such as wash solution inlet port 240 of the fluid multiplexer unit 200 of fig. 2. As depicted in fig. 4, a fluid interface block assembly 330 mounted on the fluid multiplexer block third face 106 has a first flexible tube set 334 and a second flexible tube set 336. For each fluid multiplexer block unit of fluid multiplexer block 100, each tube of flexible tube set 334 is connected to a corresponding primary waste outlet port, such as primary waste outlet port 266 of fluid multiplexer unit 200 of fig. 2, and each tube of flexible tube set 336 is connected to a corresponding calibration solution inlet port, such as calibration solution inlet port 234 of fluid multiplexer unit 200 of fig. 2. Thus, each tube of the flexible tube set 324 can be coupled and sealed to the first set of ports on the third face 106, while each tube of the flexible tube set 326 can be coupled and sealed to the second set of ports on the third face 106. Similarly, each tube of the flexible tube set 334 can be coupled and sealed to the third set of ports on the third face 106, while each tube of the flexible tube set 336 can be coupled and sealed to the fourth set of ports on the third face 106.

Finally, as depicted in fig. 4, a fourth face 108 of the fluid-multiplexer block opposite to third face 106 of the fluid-multiplexer block has a sensor device interface inlet connector port and a sensor device interface outlet connector port for each fluid-multiplexer cell of fluid-multiplexer block 100, as illustrated in fig. 2, which depicts sensor device interface inlet connector ports 260A-260D and sensor device interface outlet connector ports 262A-262D on fourth face 108 of the fluid-multiplexer block.

As previously provided herein, once installed on the fluid multiplexer block 100, each fluid line in each set of flexible tubes is coupled and sealed to an inlet port of the fluid multiplexer unit. Fig. 5A generally shows a fluid interface block assembly 310, in which a fluid interface block 312 includes a first flexible tube set 314 and a second flexible tube set 316 connected to the fluid interface block 312. As depicted in fig. 5A, each tube of a set of flexible tubes has a flanged tube connection, such as any tube of the flexible tube set 314 having the flanged tube connection 301. Additionally, each flanged pipe connection has an O-ring mounted thereon, such as O-ring 303 mounted on flanged pipe connection 301 of fig. 5A. Fig. 5B generally illustrates the sealing of each tube of a set of tubes of a fluidic interface assembly, such as fluidic interface assembly 310 of fig. 4, in each corresponding inlet port of a fluidic multiplexer unit, such as fluidic multiplexer unit 200 of fig. 2. Fig. 5B is a partial cross-sectional view of a fluid multiplexer unit, such as fluid multiplexer unit 200 of fig. 2, with fluid interface block 312 mounted thereon. Fig. 5B depicts a partial cross-sectional view through the fluid interface block 312 and fluid interface side 204, showing the flange 301 fitted into the reagent inlet port 222 and the connection between the flexible tubing set 314 and the reagent inlet port 222 effectively sealed by the O-ring 303. Similarly, fig. 5B shows the flange 305 fitting into the nucleotide reagent inlet port 228 and the connection between the flexible tubing set 316 and the nucleotide reagent inlet port 228 effectively sealed by the O-ring 307.

FIG. 6 is a schematic representation of a fluid control system 1000 that generally demonstrates a sequencing system in accordance with the teachings of the present invention. As depicted in fig. 6, fluid control system 1000 has a pneumatic control system 500 and a fluid process control system, such as fluid process manifold 600 and fluid distribution manifold assembly 700.

In this regard, the pneumatic control system 500 is in fluid communication with a first scrubbing solution container 520 through a first pneumatic inlet line controlled via valve 501, a second scrubbing solution container 522 through a second pneumatic inlet line 504 controlled via valve 503, and a cleaning solution container 524 through a third pneumatic inlet line 506 controlled via valve 505. Similarly, the pneumatic control system 500 is in fluid communication with the bulk container assembly 670 through a fourth pneumatic inlet line 508 controlled via a valve 507. With respect to fluid system control, the pneumatic control system 500 is in fluid communication with the fluid handling manifold 600 through a fifth pneumatic inlet line 510 controlled via a valve 509. Finally, the pneumatic control system 500 is in fluid communication with the clamp manifold 800 through a sixth pneumatic inlet line 512 controlled via a valve 511.

As will be provided in greater detail later herein, the pneumatic control system 500 and clamp manifold 800, along with the flow rate sensor 610, constitute a fluid regulation and control system that provides regulation and control, for example, between various input source vessels and output vessels. In this regard, various embodiments of fluid conditioning and control systems taught by the present invention may provide a defined and controllable pressure differential between input source vessels and output vessels containing various solutions for sequencing runs, which may include various nucleotide reagents for analysis, calibration solutions, wash solutions, and cleaning solutions. Thus, the fluid conditioning and control system of the present teachings can provide a defined and controllable pressure differential between the various solutions in the input source containers, such as the first wash solution container 520, the second wash solution container 522, the cleaning solution container 524, and the bulk container assembly 670, and the output container, such as the waste container 550. Thus, the defined and controlled pressure differential provides a defined and controlled flow rate of various reagents and solutions used in the analytical process through the various fluidic circuits of the fluidic control system 1000. The flow rate may comprise: a rate of about 15 microliters/second for single lane sensor device flow; about 30 microliters/second for primary waste flow; for the combination of single lane sensor device and main waste flow, 45 microliters/second; 180 microliters/second for the complete sensor device and primary waste flow; and over 300 microliters/second during system cleaning operations.

The fluid handling manifold 600 provides control over the distribution of various solutions used in cleaning and filling operations. As depicted in fig. 6, fluid handling manifold 600 has a fluid handling manifold line 620 in fluid communication with flow rate sensor 610. In accordance with the teachings of the present invention, flow rate sensor 610 provides dynamic inputs to pneumatic control system 500 for calibrating a defined flow rate of the various fluid circuits of fluid control system 1000 using clamp manifold 800 and for providing a defined flow volume when filling the various containers in bulk container assembly 670, such as nucleotide reagent container 673-679 and calibration solution container 671. With respect to the liquid input source, the first wash solution container 520 is in fluid communication with the fluid treatment manifold 600 through a first wash solution outlet line 530, while the second wash solution container 522 is in fluid communication with the fluid treatment manifold 600 through a second wash solution outlet line 532, and the cleaning solution container 524 is in fluid communication with the fluid treatment manifold 600 through a calibration solution outlet line 534.

To provide sufficient reagent volumes for massively parallel processing performed during next generation sequencing, fluid control system 1000 is configured to provide large volumes of various reagents and solutions for use in various analytical processes performed using sensor devices, such as sensor device 10 of fig. 3A. To provide perspective, the volume of the various reagents and solutions provided may be between, for example, 10,000 and 100,000 times greater than the flow cell volume of the sensor device. In this regard, the reagent concentrate cartridge 660 is in fluid communication with the wash solution containers 520 and 522. As used herein, a wash solution is a water-based solution that stabilizes the electrolyte composition, which can be used as a solvent in preparing calibration solutions and sequencing reagents during calibration of the sensor device prior to a sequencing run, for cleaning operations of the fluid manifold block as previously provided herein, and for constantly updating the electrolyte solution around the reference electrode. The reagent concentrate cartridge 660 may contain a calibration solution concentrate container 661, while the remaining concentrate is the nucleotide reagent concentrate. For example, the first nucleotide reagent concentrate container 663 may contain a deoxyguanosine triphosphate (dGTP) reagent concentrate, while the second nucleotide reagent concentrate container 665 may contain a deoxycytidine triphosphate (dCTP) reagent concentrate, and the third nucleotide reagent concentrate container 667 may contain a deoxyadenosine triphosphate (dATP) reagent concentrate, while the fourth nucleotide reagent concentrate container 669 may contain a deoxythymidine triphosphate (dTTP) reagent concentrate. The reagent concentrate cartridge 660 is also in fluid communication with the bulk container assembly 670.

Thus, each container of the bulk container assembly 670 contains a large volume of calibration solution for determining the time signal generated spatially across the sensor device as well as dNTP reagents used in high-throughput sequencing. For example, each container in the bulk container assembly 670 may have a volume of between about 200 ml and about 300 ml to load between about 140 ml and about 160 ml of the bulk calibration solution and the diluted volume of each bulk nucleotide reagent, where about 35 ml to about 135 ml may be used for a sequencing run, depending on the end-user settings of the sequencing run. In general, the volume of the bulk container may be selected, for example, by applying a suitable dilution factor for the volume of concentrated reagent in the reagent concentrate cartridge and allowing sufficient headspace to contain a pressure sufficient to provide a defined and controlled flow rate of the various solutions used in the analysis process through the various fluidic circuits of the fluidic control system 1000.

With respect to the wash solution containers 520 and 522, the volume of wash solution used for a sequencing run, for example, may be up to about 2.5 liters depending on the end user settings of the sequencing run, such that each wash solution container may be about 1.5 to about 2 liters, given that during the sequencing run, the wash solution may be used as a solvent in preparing various reagents and for constantly updating the electrolyte solution around the reference electrode. In general, the volume of the wash solution container may be selected, for example, by including and applying appropriate time and flow rate factors such as time according to the flow rates previously provided herein and allowing sufficient headspace to accommodate a pressure sufficient to provide a defined and controlled flow rate of the wash solution through the fluid control system 1000, taking into account multiple uses of the wash solution during unattended operation of the sequencing system.

Finally, with respect to the cleaning solution container 524, between 500 ml and about 700 ml of cleaning solution may be used for various cleaning procedures using cleaning solution, such that the cleaning solution container 524 may have a volume between about 600 ml and about 800 ml that may provide sufficient headspace to contain a pressure sufficient to provide a defined and controllable flow rate of cleaning solution through the fluid control system 1000.

Before the sequencing run is initiated, a bulk preparation of the calibration solution and nucleotide reagents may be performed. With respect to the bulk preparation of the calibration solution, with the valves 602 and 621 of the fluid treatment manifold 600 open and all other fluid treatment manifold valves closed, the wash solution in the wash solution container 520 may flow through the wash solution outlet line 530 into the fluid treatment manifold line 620 and through the calibration concentrate line 631 into the calibration solution concentrate container 661, and then through the calibration solution inlet line 641 into the calibration solution container 671. The wash solution may continue to flow through the calibration solution concentrate container 661 to the calibration solution container 671 to reach the predetermined fill volume of the calibration solution container 671 at which time the calibration solution concentrate container 661 has effectively drained the calibration solution concentrate.

Next, bulk preparation of the first nucleotide reagent may be performed by having valves 602 and 623 of the fluid processing manifold 600 open and all other fluid processing manifold valves closed, such that the wash solution in the wash solution container 520 may flow into the fluid processing manifold line 620 and into the first nucleotide reagent concentrate container 663 through the first nucleotide reagent concentrate line 633 and then into the first nucleotide reagent container 673 through the first nucleotide reagent inlet line 643 until the first nucleotide reagent container 673 is filled.

After the bulk preparation of the first nucleotide reagent is complete, the bulk preparation of the second nucleotide reagent may be performed by having valves 602 and 625 of the fluid handling manifold 600 open and all other fluid handling manifold valves closed, such that the wash solution in the wash solution container 520 may flow into the fluid handling manifold line 620 through the wash solution outlet line 530 and into the second nucleotide reagent concentrate container 665 through the second nucleotide reagent concentrate line 635 and then into the second nucleotide reagent container 675 through the second nucleotide reagent inlet line 645 until the second nucleotide reagent container 675 is filled.

After the bulk preparation of the second nucleotide reagent, the bulk preparation of the third nucleotide reagent may be performed by having valves 602 and 627 of the fluid processing manifold 600 open and all other fluid processing manifold valves closed, such that the wash solution in the wash solution container 520 may flow into the fluid processing manifold line 620 through the wash solution outlet line 530 and into the third nucleotide reagent concentrate container 667 through the third nucleotide reagent concentrate line 637 and then into the third nucleotide reagent container 677 through the third nucleotide reagent inlet line 647 until the third nucleotide reagent container 677 is filled.

Finally, bulk preparation of the fourth nucleotide reagent may be performed by having valves 602 and 629 of the fluid treatment manifold 600 open and all other fluid treatment manifold valves closed, such that the wash solution in the wash solution container 520 may flow through the wash solution outlet line 530 into the fluid treatment manifold line 620 and through the fourth nucleotide reagent concentrate line 639 into the fourth nucleotide reagent concentrate container 669 and then through the fourth nucleotide reagent inlet line 649 into the fourth nucleotide reagent container 679 until the fourth nucleotide reagent container 679 is filled.

With sufficient calibration solutions and nucleotide reagents prepared for high throughput next generation sequencing-by-synthesis runs, the fluid distribution manifold assembly 700 can control the distribution of various solutions through the fluid multiplexer block and into the sensor device and finally into the waste container. For example, the fluid distribution manifold assembly 700 may control the distribution of various solutions in the bulk container assembly 670 and wash solutions in the wash solution containers 520, 522 through the fluid multiplexer block 100 and into the sensor apparatus 10 and finally into the waste 530 through the sensor apparatus waste line 324 or the main waste line 334 (see, e.g., fig. 4). More specifically, during a sequencing run, the fluid distribution manifold assembly 700 may control the order in which nucleotide reagents (i.e., dNTP reagents) in the bulk container assembly 670 flow through the fluidic multiplexer block 100 and into the sensor device 10 and then finally to waste. The flow of dNTP reagents through a flow cell, such as one or more selected lanes of flow cell 6 of sensor device 10 of fig. 3A, can be performed in any determined flow order during a sequencing run.

As depicted in fig. 6, fluid distribution manifold assembly 700 may include solution distribution manifold 702, reagent distribution manifold 712, and reagent distribution manifold 722. Heater block 750 of fig. 6 may be in contact with flexible tube sets 304, 306, 314, 316, 326, and 336 to ensure that the solution and reagents have a uniform temperature before flowing through fluid multiplexer block 100 and sensor apparatus 10.

The solution distribution manifold 702 may have a first set of valves in a valve block 704 that may individually control fluid communication between the fluid treatment manifold line 620 and the flexible tube set 326. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 326 is connected to a corresponding wash solution inlet port of a corresponding fluid multiplexer unit, such as the wash solution inlet port 240 of the fluid multiplexer unit 200 of fig. 2. Additionally, the solution distribution manifold 702 may have a second set of valves in a valve block 706 that may individually control fluid communication between the calibration solution line outlet 651 and the flexible tube set 336. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 336 is connected to a corresponding calibration solution inlet port of a corresponding fluid multiplexer unit, such as the calibration solution inlet port 234 of the fluid multiplexer unit 200 of fig. 2.

To provide for flow of wash solution through a selected lane of the multi-lane sensor device or through multiple selected lanes, either valve 602 or 604 of fluid processing manifold 600 may be opened while all other valves in fluid processing manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 704 of the solution distribution manifold 702 may be opened, so wash solution from either vessel 520 and 522 may flow through either wash solution outlet line 530 or 532 into the fluid treatment manifold line 620 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, respectively, depending on the selection of the valve in the valve block 704. To provide for the flow of calibration solution through one selected lane of the multi-lane sensor device or through multiple selected lanes, all valves in fluid handling manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 706 of the solution distribution manifold 702 may be open, so calibration solution from the calibration solution container 671 may flow into the calibration solution line outlet 651 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, depending on the selection of valves in the valve block 706.

The reagent distribution manifold 712 can have a first set of valves in a valve block 714 that can individually control fluid communication between the first nucleotide reagent outlet line 653 and the flexible tubing set 304. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 304 is connected to a corresponding first nucleotide reagent inlet port of a corresponding fluid multiplexer unit, such as the first nucleotide reagent inlet port 210 of the fluid multiplexer unit 200 of fig. 2. Additionally, the reagent distribution manifold 712 may have a second set of valves in a valve block 716 that may individually control fluid communication between the second nucleotide reagent outlet line 655 and the flexible tubing set 306. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 306 is connected to a corresponding second nucleotide reagent inlet port of a corresponding fluid multiplexer unit, such as the second nucleotide reagent inlet port 216 of the fluid multiplexer unit 200 of fig. 2.

For example, to provide flow of the first nucleotide reagent through one selected lane of the multi-lane sensor device or through multiple selected lanes, all valves in fluid processing manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 714 of the distribution manifold 712 can be opened so that the first nucleotide reagent from the first nucleotide reagent container 673 can flow into the first nucleotide reagent outlet line 653 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, depending on the selection of the valve in the valve block 714. To provide for the flow of the second nucleotide reagent through one selected lane of the multi-lane sensor device or through multiple selected lanes, all valves in fluid handling manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 716 of the distribution manifold 712 may be opened so that the second nucleotide reagent from the second nucleotide reagent container 675 may flow into the second nucleotide reagent outlet line 655 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, depending on the selection of valves in the valve block 716.

The reagent distribution manifold 722 may have a first set of valves in a valve block 724 that can individually control fluid communication between the third nucleotide reagent outlet line 657 and the flexible tubing set 314. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 314 is connected to a corresponding third nucleotide reagent inlet port of a corresponding fluid multiplexer unit, such as the third nucleotide reagent inlet port 222 of the fluid multiplexer unit 200 of fig. 2. Additionally, the reagent distribution manifold 722 may have a second set of valves in a valve block 726 that may individually control fluid communication between the fourth nucleotide reagent outlet line 659 and the flexible tube set 316. As depicted for the fluid multiplexer block assembly 110 of fig. 4, each tube of the flexible tube set 316 is connected to a corresponding fourth nucleotide reagent inlet port of a corresponding fluid multiplexer unit, such as the fourth nucleotide reagent inlet port 228 of the fluid multiplexer unit 200 of fig. 2.

For example, to provide for flow of the third nucleotide reagent through one selected lane of the multi-lane sensor device or through multiple selected lanes, all valves in fluid processing manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 724 of the distribution manifold 722 may be open, so the third nucleotide reagent from the third nucleotide reagent container 677 may flow into the third nucleotide reagent outlet line 657 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, depending on the selection of the valve in the valve block 724. To provide for the flow of the fourth nucleotide reagent through one selected lane of the multi-lane sensor device or through multiple selected lanes, all valves in fluid handling manifold 600 are closed. One valve, all four valves, or any combination of valves in the valve block 726 of the distribution manifold 722 may be open, so that the fourth nucleotide reagent from the fourth nucleotide reagent container 679 may flow into the fourth nucleotide reagent outlet line 659 for distribution by the corresponding fluid multiplexer block unit to one lane, all lanes, or any combination of lanes, depending on the selection of the valve in the valve block 726. Although a flow order is provided in this example, it should be noted that dNTP flow order can be performed in any defined flow order during a sequencing run.

A schedule of various cleaning procedures for all fluid components in fluid control system 1000 may be executed.

For example, a run-to-run clean that provides a flush of used lanes may be performed to clean previously used lanes. For example, once the sequencing run has been completed and the sensor device still has unused lanes, the fluid control system 1000 of FIG. 6 may flush the cleaning solution from the cleaning solution container 524 through fluid passage 632-. After a sufficient volume of cleaning solution has been flushed through the system, the sensor device 10 may be further processed for another sequencing run.

Additionally, a cleaning schedule for all fluid components in the fluid control system 1000 may be performed. Such cleaning is typically performed after all lanes of the sensor device have been sequenced or before a new sensor device is installed on the system. The cleaning may be performed with the depleted reagent concentrate cartridge and the used multi-lane sensor device in place. With valve 606 open, each of valves 623-629 of fluid handling manifold 600 may be opened in sequence and all valves in a set of valves of a corresponding valve block of fluid distribution manifold assembly 700 may be opened. Where such flow paths are performed sequentially for each fluidic path of the calibration solution and each nucleotide reagent, the cleaning solution from the cleaning solution container 524 may flow sequentially through each fluidic component of the fluidic control system 1000 to the waste container 550. Finally, a drying procedure is performed to prepare the system for the next use. For the drying procedure, valves 602, 604, and 606 are closed, and all other valves of fluid handling manifold 600 are open, and all valves of fluid distribution manifold assembly 700 are open. In this configuration, clean dry air passes through the fluid handling components of the fluid control system 1000 to drive the remaining liquid to the waste container 550.

As previously provided herein, the pneumatic control system 500 and clamp manifold 800, along with the flow rate sensor 610, constitute a fluid regulation and control system that provides regulation and control, for example, between various input source vessels and output vessels. In this regard, various embodiments of fluid conditioning and control systems taught by the present invention may provide a defined and controllable pressure differential between input source vessels and output vessels containing various solutions for sequencing runs, which may include various nucleotide reagents for analysis, calibration solutions, wash solutions, and cleaning solutions. Thus, the fluid conditioning and control system of the present teachings can provide a defined and controllable pressure differential between the various solutions in the input source containers, such as the first wash solution container 520, the second wash solution container 522, the cleaning solution container 524, and the bulk container assembly 670, and the output container, such as the waste container 550.

Regarding the components of the fluid regulation and control system, for various embodiments of the fluid control system 100, the clamp manifold 800 may contain eight clamp regulators, each clamp regulator configured as described in US 9,375,716. These devices operate essentially as three-port pressure followers with an input fluid port, an output fluid port, and a control pneumatic port. With a defined waste line fluid resistance connected between the pinch regulator output fluid port and the waste container 550, the pressure on the output fluid port will be approximately equal to the pressure on the pinch regulator control pneumatic port, regardless of the pressure on the input fluid port. The flow rate through the pinch regulator will then be equal to the output fluid port pressure divided by the waste line fluid resistance. To accurately calibrate the flow rate, the fluid control system 1000 is configured to allow the wash solution to flow to the desired clamp regulator on the clamp manifold 800. A known set of pneumatic pressures is applied to each clamp regulator pneumatic control port and a flow sensor 610 measures a precise flow rate corresponding to the pneumatic control pressure. Then, for each pinch regulator, a table of flow rates versus pneumatic control pressures is stored, which the instrument software can use to accurately deliver any desired flow rate.

Fig. 7 is a rear isometric view generally showing a fluid multiplexer block clamp assembly 150. As depicted in fig. 7, fluid-multiplexer block clamp assembly 150 includes a fluid-multiplexer block clamp 400 in which fluid-multiplexer block assembly 110 is mounted. The fluid multiplexer block clamp 400 may include an electrode connection mounting plate 410 mounted on one side 342 of the electrode adapter fluid interface block 340. The electrode connection mounting plate 410 enables connection of electrical leads 412 and ground leads 414 to the electrode adapter fluid interface block 340 of the fluid multiplexer block clamp assembly 150. Fluid multiplexer block clamp 400 also includes shoulder screws 420, 422, and 424 and a fourth shoulder screw placed below shoulder screw 424 and opposing shoulder screw 422. The force on the shoulder screw of the fluid multiplexer block clamp 400 is set to provide a four degree movement to the fluid multiplexer block mounted therein to provide flexibility in interfacing the fluid multiplexer block with the multi-lane sensor device.

With respect to the fluid-multiplexer block assembly 110 installed in the fluid-multiplexer block clamp 400, as depicted in fig. 7, the orientation of the fluid-multiplexer block 100 and the fluid block connection shows the fluid interface block 312 mounted to the fluid-multiplexer block 100 at the top of the fluid-multiplexer block clamp assembly 150, while the fluid interface block 302 is mounted to the fluid-multiplexer block 100 at the bottom of the fluid-multiplexer block clamp assembly 150. As depicted in fig. 7, the orientation of the first and second flexible tube sets 314, 316 likewise emanates from the top of the fluid multiplexer block clamp assembly 150, while the orientation of the first and second flexible tube sets 304, 306 likewise emanates from the bottom of the fluid multiplexer block clamp assembly 150. Similarly, fluid interface block 332 is mounted to fluid multiplexer block 100 below the back of fluid multiplexer block clamp assembly 150 and electrode adapter fluid interface block 340. As depicted in fig. 7, the orientation of first flexible tube set 334 and second flexible tube set 336 likewise emanate from the back of fluid multiplexer block clamp assembly 150. Finally, fluid interface block 322 is mounted to fluid multiplexer block 100 over the back of fluid multiplexer block clamp assembly 150 and electrode adapter fluid interface block 340. As depicted in fig. 7, the orientation of the first flexible tube set 324 and the second flexible tube set 326 likewise emanates from the top of the electrode adapter fluid interface block 340.

Fig. 8A and 8B are cross-sectional views that generally show the orientation of the fluid-multiplexer block 100 when it is installed in a fluid-multiplexer block clamp assembly and the integration of electrodes into the fluid-multiplexer cell. With respect to the orientation of the fluid-multiplexer block 100 when it is installed in the fluid-multiplexer block jig assembly, as depicted in fig. 8A, the orientation is a 90 ° counterclockwise rotation of the fluid-multiplexer block assembly 110 of fig. 4. In this regard, the fluid multiplexer block first face 102 is depicted as having a fluid interface block 302 mounted thereon and having a first flexible tube set 304 and a second flexible tube set 306 connected to the fluid interface block 302, while the fluid multiplexer block second face 104, opposite the fluid multiplexer block first face 102, is depicted as having a fluid interface block 312 mounted thereon and having a first flexible tube set 314 and a second flexible tube set 316 connected to the fluid interface block 312. Similarly, the fluid-multiplexer block third face 106 is depicted as having a fluid interface block 332 mounted thereon and having a first flexible tube set 334 and a second flexible tube set 336 connected to the fluid interface block 332. Additionally, the fluid multiplexer block third face 106 is depicted as having an electrode adapter fluid interface block 340 mounted thereon. As depicted in fig. 8A, the fluid interface block 322 is mounted on the electrode adapter fluid interface block 340 such that the first and second flexible tubing sets 324 and 326 connected to the fluid interface block 332 are in fluid communication with the electrode adapter fluid interface block inlet channels 342 and 344, respectively. In this regard, the electrode adapter fluid interface block 340 is mounted to the fluid multiplexer block third face 106 such that the electrode adapter fluid interface block inlet channels 342 and 344 are coupled and sealed to the sensor device waste outlet port 264 and the wash solution inlet port 240, respectively. Finally, as depicted in fig. 8A, the fluid multiplexer block fourth face 108 has a sensor device interface inlet connector port 260 and a sensor device interface outlet connector port 262. As previously provided herein, fluid multiplexer block fourth face 108 has a corresponding set of sensor device interface inlet connector ports and sensor device interface outlet connector ports for each fluid multiplexer unit of fluid multiplexer block 100. As will be provided in more detail later herein, the sensor device interface inlet connector port 260 and the sensor device interface outlet connector port 262 are coupled and sealed to the inlet port and the outlet port, respectively, of the multi-lane sensor device.

With respect to the electrode connections provided to each fluid multiplexer unit, which provide a constant, stable reference electrode voltage to the multi-lane sensor device, FIG. 8A depicts a cross-sectional view of an electrode adapter fluid interface block 340 with an electrode connection mounting plate 410 mounted thereon. Fig. 8A depicts electrodes 275 in enlarged holes in a cross section of the electrode adapter fluid interface block inlet channel 344 that provide fluid communication through the electrode adapter fluid interface block inlet channel 344 in fluid communication with the wash solution channel 242. The electrodes 275 are electrically coupled to a voltage source connected to the electrode connection mounting board 410 by electrical leads 412 and ground leads 414 (see fig. 7). As previously provided herein, the second flexible tubing set 326 is in fluid communication with a source of wash solution that stabilizes the electrolyte composition. Thus, the electrodes 275 are in a fluid environment that provides a constant, stable reference electrode voltage to the multi-lane sensor device. Fig. 8B is an expanded view of the electrode adapter fluid interface block 340 depicting electrodes 275 in enlarged holes in the cross-section of the electrode adapter fluid interface block inlet channel 344 and depicting the electrodes 275 coupled to the electrode connection mounting plate 410. As depicted in fig. 8A, the electrode adapter fluidic interface block inlet channel 344 is in fluidic communication with the flexible tubing set 326 and is coupled and sealed to the wash solution inlet port 240, which is in fluidic communication with the wash solution channel of the fluid multiplexer unit, as previously provided herein. For example, as depicted in fig. 2, wash solution inlet port 240 is in fluid communication with wash solution channel 242 of fluid multiplexer unit 200.

Fig. 9 is a front isometric view generally showing a fluid multiplexer block clamp assembly 150 including a fluid multiplexer block clamp 400 in which the fluid multiplexer block assembly 110 is mounted. As depicted in fig. 9, fluid multiplexer block fourth face 108 of fluid multiplexer block 100 has a sensor device interface inlet connector port 260A-260D and a sensor device interface outlet connector port 262A-262D, respectively, for each fluid multiplexer cell 200A, 200B, 200C, and 200D. The first alignment notch 107A of the first fluid manifold unit 200A and the second alignment notch 107B of the fourth fluid manifold unit 200D are configured to facilitate the alignment and sealing process of the fluid multiplexer block clamp assembly 150 with respect to the multi-lane sensor device. As previously provided herein, the fluid multiplexer block clamp 400 provides four degrees of movement to a fluid multiplexer block mounted therein to provide flexibility in interfacing the fluid multiplexer block with a multi-lane sensor device. Additionally, the first alignment recess 107A and the second alignment recess 107B are configured to provide self-alignment of the multi-lane sensor device with the multi-lane sensor device such that sealing of the sensor device interface inlet connector ports and the sensor device interface outlet connector ports, such as the sensor device interface inlet connector ports 260A-260D and the sensor device interface outlet connector ports 262A-262D, may be performed with respect to the respective inlet and outlet ports of the multi-lane sensor device.

Figure 10 is a cross-sectional view generally showing a fluid multiplexer block assembly 110 mounted to a multi-lane sensor device 10, such as the multi-lane sensor device 10 schematically depicted in figure 3B. As depicted in fig. 10, the multi-lane sensor device 10 mounted to the sensor device mounting and positioning assembly 450. The location of the fluid interface blocks 302, 312, 322, and 332 of the fluid-multiplexer block assembly 110 is also evident in the cross-sectional view of fig. 10. When the fluid multiplexer block assembly 110 is installed to the multi-lane sensor device 10, the coupling and sealing of each sensor device interface inlet connector port and each sensor device interface outlet connector port to each corresponding sensor device inlet port and each sensor device outlet port is performed separately for each lane of the multi-lane sensor device. In this regard, fig. 11 is an expanded isometric view generally illustrating the mounting and sealing of the fluid multiplexer block to the multi-lane sensor device. As depicted in fig. 11, the sensor device 10 has lanes 4A to 4D, each lane having an inlet port and an outlet port, as illustrated for lane 4A having inlet port 3A and outlet port 5A. In fig. 11, the juxtaposition of the first alignment pin 12A of the sensor device 10 and the first alignment notch 107A of the fluid-multiplexer block 100 illustrates how the complementary pairs engage to align the sensor device 10 with the fluid-multiplexer block 100. Further, fig. 11 depicts how each inlet port and each outlet port of sensor device 10 may be coupled and sealed to each corresponding sensor device interface inlet connector port and each sensor device interface outlet connector port of fluid multiplexer block 100. In fig. 11, this is illustrated for lane 4A, where the juxtaposition of first inlet port 3A and sensor device interface inlet connector port 260A and first outlet port 5A and sensor device interface outlet connector port 262A may be coupled and sealed to each once sensor device 10 and fluid multiplexer block 100 have fully engaged each other. This compressive coupling and sealing provides for a quick decoupling of the fluid multiplexer block 100 from the sensor device 10.

FIG. 12 is a block diagram generally illustrating a sequencing system, which may be a sequencing system incorporating a sample preparation platform, in accordance with the teachings of the present invention. As depicted in fig. 12, the sequencing system 2000 can include a controller 2002 in communication with a sample preparation spacer 2004, a loading station 2006, and a sequencing station 2008. The sample preparation spacer 2004 may include a pipetting robot 2012, which may be a three-axis pipetting robot. The pipetting robot 2012 can access the sample 2014, the reagents and solutions 2016, the thermal cycler 2018, and other devices 2020, such as a magnetic separator or centrifuge. A target sequence of a sample to be analyzed on the sequencing system 2000 can be prepared at the sample preparation spacer 2004 and can then be provided to the loading station 2006. For example, sample preparation spacer 2004 may provide for library preparation of a sample to be analyzed as well as preparation of target sequences from a library of samples that may be used to prepare particles or beads. A sample of such particles or beads can then be provided to a loading station 2006 for loading onto a sensor device, such as the sensor device 10 of fig. 3A.

Once loaded, the sensor device may be transported to a sequencing station 2008 using a slide mechanism 2007 that can move the sensor device from a loading position to a sequencing position. The sequencing station 2008 may include fluidic and electronic interfaces to automatically process samples loaded on the sensor devices during a sequencing run. The container cabinet 2010 may contain containers that hold various reagents and solutions used in a sequencing run, and contain various waste containers. The data collected from the sensing devices can be provided to a sequencing computer 2022 that can perform base calling, read alignment, and variant calling.

The controller 2002 may further be in communication with a user interface such as a monitor, keyboard, mouse, touch screen, or any combination thereof, as well as other interfaces such as the user interface 2024 of fig. 12. Further, the controller 2002 may communicate with a network interface that may access a local area network, a wide area network, or a global network. Network interface 2026 may be a wired interface or a wireless interface using various standard communication protocols. The sequencing system 2000 may be powered by a power supply 2028.

FIG. 13 is a perspective view of a sequencing system that generally demonstrates the teachings of the present invention. The sequencing system 2500 of fig. 13 can be a sequencing system having various components as described for the sequencing system 2000 of fig. 12. Sequencing system 2500 can comprise an upper portion 2502 and a lower portion 2504. The upper portion 2502 may contain a door 2506 for accessing the sample preparation compartment 2504 upon which samples, reagent containers and other consumables to be analyzed may be placed, such as described with respect to fig. 12. Lower portion 2504 may contain a container cabinet, such as container cabinet 2010 of fig. 12. Additionally, various embodiments of the sequencing system, such as the sequencing system 2500, may include a user interface, such as a touch screen display 2508.

Fig. 14 generally illustrates a container cabinet 2510, which can be a component of a sequencing system, such as the sequencing system 2000 of fig. 12 and the sequencing system 2500 of fig. 13. The container cabinet 2510 may be used to manage the fluid processing of the sequencing system. For example, as depicted in fig. 13, the container cabinet 2510 includes a kit loading interface 2512 for loading a reagent concentrate cartridge, such as the reagent concentrate cartridge 660 of fig. 6. Further, the container cabinet 2510 may contain various containers for holding reagents and solutions. For example, the wash solution and the cleaning solution may be contained in the containers of the first container assembly 2514 of fig. 12, as depicted for containers 520, 522, and 524 of fig. 6. Further, the bulk nucleotide reagents and the bulk calibration solution may be contained in the container of the second container assembly 2516 of fig. 14, such as depicted for the bulk container assembly 670 of fig. 6. As depicted in fig. 6, the second container assembly 2516 of fig. 14 can be in fluid communication with the fluid multiplexer block 100 as shown in fig. 6, as through the flexible tube sets 304, 306, 314, 316, and 336. The fluidic multiplexer block 100 as shown in fig. 6 is fluidically coupled to a sensor device 10, which may be an ISFET device for sequencing-by-synthesis. Additionally, the container cabinet may contain various sample preparation waste containers, sensor waste containers, and primary waste containers for collecting effluent. For example, the first waste container 2518A may collect effluent generated during sample preparation, such as effluent generated from the sample preparation septum 2004 of the sequencing system 2000 of fig. 12 and the sample preparation septum 2504 of fig. 13. Additionally, the secondary waste container 2518B of fig. 14 may collect effluent generated from a fluidic system, such as the fluidic system 100 of fig. 6, for example, during a sequencing run, such as through the sensor device waste line 324 and the primary waste line 334 of fig. 6.

Fig. 15 generally illustrates a flow chart of a method for automated fluidic system workflow for an automated sequencing system. The method 1500 of fig. 15 can be used on a sequencing system, such as the sequencing system 2000 of fig. 12 and the sequencing system 2500 of fig. 13, which can include an automated fluid control system, such as the fluid control system 1000 of fig. 6. For the method 1500 of fig. 15, once loaded with a sample of particles or beads that have been prepared for sequencing, the sensor device can be automatically moved from the loading position to the sequencing position, e.g., using the slide mechanism 2007 of fig. 12.

Before the automated sequencing run is initiated, a bulk calibration and preparation of nucleotide reagents can be performed, as indicated at step 1502 of fig. 15. As previously described, the end user may select one lane in any location of the sensor device to be used individually during a run, all lanes to be used simultaneously during a run, or any combination of lanes to be used simultaneously during a run, such that the sensor device may be used for more than one sequencing run. Thus, since the sensor device can be used for more than one sequencing run, the end user can prepare large volumes of bulk calibration solutions and nucleotide reagents with a new sensor device installed. With respect to the workflow at step 1502, as a non-limiting example, when a new sensor device is used, an end user may insert a new reagent concentrate cartridge, such as reagent concentrate cartridge 660 of fig. 6, into a kit loading interface 2512 of a sequencing system, such as sequencing system 2500 of fig. 13. Then, as part of the predictive sequencing step 1502, the preparation of the bulk calibration solution and dNTP reagents is automatically performed in order to provide bulk volumes of the calibration solution and dNTP reagents for massively parallel processing performed by automated sequencing systems, such as the sequencing system 2000 of fig. 12 and the sequencing system 2500 of fig. 13. As depicted in fig. 6 and previously described herein, the concentrate from each concentrate container, such as the concentrate container 661-669 of the reagent concentrate cartridge 660 of fig. 6, is precisely diluted into the container 671-679 of each respective bulk container, such as the bulk container assembly 670 of fig. 6. In various embodiments, the bulk container assembly 670 as depicted in fig. 6 may be an assembly of conical containers as depicted in the second container assembly 2516 of fig. 13.

With respect to the automated fluidic system workflow of step 1504, the second predictive sequence step involves priming of the fluidic circuit. For example, referring to FIG. 6, the priming fluid pathway may comprise a pathway that provides fluid communication between the wash solution container 520-. Finally, the fluid pathways from the fluid multiplexer block are in fluid communication with the sensor unit waste and the primary waste, such as through the sensor unit waste line 324 and the primary waste line 334 described with respect to and depicted in fig. 4, 6, and 7. In accordance with the teachings of the present invention, a priming step 1504 can be performed to prepare the sequencing system for a sequencing run by appropriately filling all of the lanes and ensuring that all of the lanes are bubble free and priming a fluid multiplexer block, such as the fluid multiplexer block 100 of FIG. 6, in preparation for starting the sequencing run.

In the exemplary automated workflow of step 1504, dNTP reagent priming may be performed on the fluid pathway to prime the pathway with a sequential dNTP reagent flow through one or more of the active lanes of the sensor device designated by the end user as a sequencing run. For example, with respect to the sensor apparatus 10 of FIG. 3A, the end user may select any one of the four lanes to be used individually during the run, all four lanes to be used simultaneously in any order during the run, or any combination of lanes to be used simultaneously in any order during the run. In this regard, if the end user selects any single lane of the sensor device for a sequencing run, dNTP reagent perfusion will be performed for the selected active lane, whereas if all lanes are selected to run in any order, dNTP perfusion and filling will be performed for all active lanes, and finally, if 2 to 3 lanes are selected to run in any order, 2-3 active lanes selected will be perfused and filled. For example, referring to FIG. 6, reagent priming of the fluid pathway may comprise providing fluid communication between wash solution container 520-522 and fluid pathway 632-639 and, ultimately, fluid pathways 642-649 and 652-659 in fluid communication with one or more of the selected active lanes of sensor device 10 through fluid multiplexer block 100. Finally, reagents flow from the fluidic multiplexer block 100 to the sensor unit waste and to the primary waste, for example, through the sensor unit waste line 324 and the primary waste line 334 of fig. 6.

After priming the fluidic system, a calibration of the sensor device may be performed at step 1506 of fig. 15 as a third pre-sequencing step. As described with respect to and depicted in fig. 3B, 8A, and 8B, the reference electrode 275 provides a constant and stable reference voltage for a sensor device, such as the sensor device 10 of fig. 3A and 3B. Prior to the automatic start of the sequencing run, an automated fluidic system, such as the fluidic control system 1000 of fig. 6, provides a flow of wash solution over the reference electrode and through the sensor device while the reference electrode provides a constant and stable voltage. Under such conditions, parameters required for the function of a sensor device, such as a chemFET sensor device or an ISFET sensor device, can be determined. Additionally, a calibration solution, such as calibration solution 671 of fig. 6, may be used to calibrate a sensor device, such as sensor device 10 of fig. 3A and 3B. In an embodiment, the calibration solution is selected to provide a sensor response, such as a pH change to the ISFET sensor device. As the calibration solution flows past over the sensor device, a calibration of the time during which a signal may be generated spatially across the sensor device during the nucleotide reagent flow may be determined.

In the case of sequencing system start-up and calibration, a sequencing run may be initiated. The sequential flow of the nucleotide solution and the wash solution during the sequencing run at step 1508 may be provided by an automated fluidic system as previously described herein with respect to the fluidic control system 1000 of fig. 6. In an example, the fluid control system 1000 can control the sequence of nucleotide reagent flow by controlling the fluid pathways between the wash solution container 520-522 and the fluid pathway 632-639, and the fluid pathways 642-649 and 652-659 that ultimately fluidly communicate with the sensor device 10 through the fluid multiplexer block 100. During a sequencing run, the sequential flow of nucleotide reagents through the end-user selected active lanes of the sensor device may be in any determined flow order and finally from the fluidic multiplexer block to the sensor device waste and to the primary waste, for example, through the sensor device waste line 324 and the primary waste line 334 of fig. 6.

Once the sequencing run has been completed and the sensor device still has unused lanes, cleaning the fluidic system between runs may be performed at step 1510. At the end of a sequencing run, an automated fluidic system such as the fluid control system 1000 of fig. 6 may provide a flow of cleaning solution from the cleaning solution container by a lane selected by an end user as an active lane during a sequencing run setup while the sensor device with the unused lane is still in a sequencing position. For example, referring to FIG. 6, cleaning the fluid pathways between sequencing runs may include providing pathways to clean fluid communication between solution container 524 and fluid pathways 632-. The fluid multiplexer block 100 is in fluid communication with one or more lanes of the sensor apparatus 10 that are already active lanes such that the cleaning solution is flushed through the used lanes and finally flushed to sensor apparatus waste and to primary waste, for example, through the sensor apparatus waste line 324 and the primary waste line 334 of fig. 6. In examples where the end user selects two lanes, e.g., lane 4A and lane 4C of fig. 3A, 3B, 6, and 11, an automated fluidic system, e.g., the fluid control system 1000 of fig. 6, may flush a cleaning solution through lanes 4A and 4C after the end of a sequencing run. After a sufficient volume of cleaning solution has been flushed through the system, the fluidic system may be depressurized to remove liquid from the pipeline so that the sensor device may be decoupled from the fluidic multiplexer block as described for and depicted in fig. 10 and 11. Once decoupled from the fluid multiplexer block, the sensor device with unused lanes may be automatically moved from the sequencing position to the loading position, for example, using the slide mechanism 2007 of fig. 12. When positioned in the loading position, the sensor device with unused lanes may be loaded with a sample of particles or beads that have been prepared for sequencing to perform a subsequent sequencing run according to a user-defined run plan. Once loaded and returned to the sequencing location, method steps 1502 through 1510 can be repeated until, for example, the sensor device has been fully used or otherwise depleted.

When the end user has fully used the sensor device or when the device is otherwise depleted, and before the system is initialized with a new sensor device, the fluid system may be cleaned as indicated at step 1512 of fig. 15 and as previously described herein with respect to fluid control system 1000 of fig. 6. Recall that with the depleted reagent concentrate cartridge and depleted sensor device in place, and with valve 606 open, each of the valves 623-629 of the fluid handling manifold 600 may be opened in sequence, and all of the valves in a set of valves of the corresponding valve block of the fluid distribution manifold assembly 700 may be opened. Where such flow paths are performed sequentially for each fluidic path of the calibration solution and each nucleotide reagent, the cleaning solution from the cleaning solution container 524 may flow sequentially through each fluidic component of the fluidic control system 1000 to the waste container 550. Finally, a drying procedure is performed to prepare the system for the next use. For the drying procedure, valves 602, 604, and 606 are closed, and all other valves of fluid handling manifold 600 are open, and all valves of fluid distribution manifold assembly 700 are open. In this configuration, clean dry air passes through the fluid handling components of the fluid control system 1000 to drive the remaining liquid to the waste container 550.

While various embodiments of the present teachings have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the teachings of the present invention. It is to be understood that various alternatives to the embodiments described herein may be employed in practicing the disclosure. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby.