Fluid supply system, method of analyzing sample, and method of assembling instrument

Abstract

The present disclosure provides a fluid supply system, a method of analyzing a sample, and a method of assembling an instrument. A fluid supply system of interest: comprising a working fluid line; a main flushing fluid line; a secondary fluid line; a flow control circuit fluidly connected to a distal end of the primary irrigation fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary irrigation fluid and the secondary fluid to pass through the outlet; and a single fluid connection configured to fluidly couple to the flow control loop, the working fluid line, and the fluid assembly of the instrument.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to the filing date of U.S. Provisional Patent Application Serial No. 63 / 546,024, filed on October 27, 2023; the disclosure of that application is incorporated herein by reference in its entirety. Background Art

[0003] Flow particle analyzers, such as flow cytometers, are well-known analytical tools that are capable of characterizing particles based on optical parameters (e.g., light scattering and fluorescence) or according to electrical properties (e.g., impedance). For example, in a flow cytometer, particles (e.g., molecules, analyte-bound beads, or individual cells) in suspension pass through a detection region (in which the particles are exposed to excitation light, typically from one or more lasers), and the light scattering and fluorescence properties of the particles are measured. The particles or their components are typically labeled with fluorescent dyes to facilitate detection, and multiple different particles or components can be detected simultaneously by labeling different particles or components with spectrally distinct fluorescent dyes. Typically, detection is performed using multiple photodetectors, one for each different dye to be detected.

[0004] In some flow cytometers, the sheath fluid is supplied to the flow cell by a pressure-driven fluid system in which the sample fluid and the sheath fluid are passed through the flow cell at a pressure above ambient pressure. The change in flow rate through the flow cell is achieved by varying the pressure in the sheath fluid reservoir, and the ratio of sample fluid to sheath fluid in the hydrodynamic flow is determined by the pressure applied in the sample source and the sheath fluid reservoir and the resistance of the fluid system supplying the sample and sheath fluid. Flow cytometers may also use vacuum-driven fluid systems in which a vacuum pump draws a vacuum downstream of the flow cell and the sample and sheath fluids are maintained at ambient pressure. To vary the rate through the flow cell, the vacuum pump draws a vacuum and the ratio of sample fluid to sheath fluid flowing through the flow cell is determined by the ratio of the resistances imposed by the path of the sample fluid and the path of the sheath fluid system. Fluid systems that provide a hydrodynamic focusing flow of a particle-containing sample fluid in the center of a particle-free sheath fluid flow typically use pressurizable tubing, connections, and seals that are required to withstand a wide range of pressure levels, particularly high and low pressures.

[0005] Pressure-based fluidics systems configured to provide sample fluid flow in a sheath fluid stream include the BD Biosciences FACSAria TM I (described in U.S. Pat. No. 6,767,188), BD Biosciences FACSAria TMII, BD Biosciences FACSAriac III, BD Biosciences FACSCantoe flow cytometer, and Bio-Rad S3e- (described in U.S. Pat. No. 9,551,637). These systems can be configured to switch between fluids so that different fluids can be provided to the flow cell at different times as needed. Typically, switching between fluids involves a manual process in which the user reconnects the fluid lines to different fluid sources. Summary of the invention

[0006] The inventors have realized that, although pressure-based fluid systems (e.g., the system proposed in U.S. Patent No. 6,767,188) constitute an improvement in fluid technology, other improvements can also be made. Specifically, it has been found that the problem in traditional fluid supply mechanisms is to prevent residues between fluids (i.e., a first fluid is left in a fluid line to be used to transport a second fluid). In certain applications, it is critical to have very low residues between multiple fluids, such as when a cleaning fluid may affect the purity of the fluid, when a cleaning fluid may affect a biological sample on an instrument, when multiple cleaning fluids or reagents are required but direct mixing of fluids may produce adverse chemical reactions, and the like. This means that the upwardly curved lines are either (1) a potential source of instrument contamination or (2) require manual removal and cleaning by the user. Since they are not completely cleaned, they also cannot ensure low fluid residues without manual intervention by the user. Therefore, a fluid supply system that is configured to be completely cleaned and have minimal residue is needed. The system and method of the present invention meet this desire.

[0007] Aspects of the present invention include a fluid supply system. The subject system includes: a working fluid line configured to deliver a working fluid therethrough from a proximal end to a distal end, and including a working fluid reservoir connector located at the proximal end, the connector configured to fluidically couple to a working fluid reservoir containing a working fluid (e.g., sheath fluid); a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end, and including a primary flush fluid reservoir connector located at the proximal end, the connector configured to fluidically couple to a primary flush fluid reservoir containing a primary flush fluid (e.g., an inert fluid); a secondary fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end, and including a primary flush fluid reservoir connector located at the proximal end, the connector configured to fluidically couple to a primary flush fluid reservoir containing a primary flush fluid (e.g., an inert fluid); A system of the invention is provided for providing a primary flushing fluid line configured to deliver a secondary fluid therethrough from a proximal end to a distal end, and including a secondary fluid reservoir connector at the proximal end, the connector being configured to be fluidically coupled to a secondary fluid reservoir containing a secondary fluid (e.g., a cleaning fluid, a reagent, etc.); a flow control circuit fluidically connected to the distal end of the primary flushing fluid line and the distal end of the secondary fluid line, and configured to selectively allow the primary flushing fluid and the secondary fluid to pass through an outlet; and a single fluid connection configured to be fluidically coupled to the flow control circuit, the working fluid line, and a fluid assembly of the instrument. The single fluid connection may include, for example, a quick-connect fluid connector or a three-way valve. The system of interest also includes a fluid supply pump operably connected to the flow control circuit and the single fluid connection. Embodiments also include a working fluid reservoir fluidically coupled to the working fluid line, and / or a pressure regulator operably connected to the working fluid reservoir and a source of pressurized air. In some cases, the system includes a working fluid supply pump operably connected to the single fluid connection. In selected versions, the system includes a primary flushing fluid reservoir fluidically coupled to the main flushing fluid line. In some embodiments, the system includes a secondary fluid reservoir fluidically coupled to the secondary fluid line. In some cases, the fluid supply system includes a plurality of secondary fluid lines. In some embodiments, the flow control circuit includes a valve group. The valve group may include, for example, a two-way valve or a three-way valve. In some cases, the two-way valve or the three-way valve is an independent valve. In other cases, the two-way valve or the three-way valve is mounted on a manifold. In selected cases, the distal end of the main flushing fluid line is fluidically coupled to the valve in the valve group that is farthest from the outlet of the flow control circuit. In some embodiments, the flow control circuit includes a multi-port selection valve.

[0008] In some embodiments, the system of the present invention also includes an instrument. The instrument may include, for example, a flow cytometer. In some such cases, the fluid assembly of the instrument is a flow cell. In some cases, the system also includes a vacuum source operably connected to the fluid assembly of the instrument. Aspects of the present invention also include methods for analyzing samples, and methods of interest include introducing samples into an instrument including a fluid supply system of the present invention, and supplying a working fluid to the instrument to analyze the sample. Aspects of the present invention also include methods for assembling instruments. Such methods include operably connecting a fluid supply system of the present invention to the fluid assembly of the instrument. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] The present invention may be better understood by reading the following detailed description in conjunction with the accompanying drawings, which include the following figures:

[0010] Figure 1 A block diagram of a fluid supply system according to some embodiments of the present invention is shown.

[0011] Figure 2 A fluid flow diagram of a fluid supply system according to some embodiments of the present invention is shown.

[0012] Figure 3 A fluid flow diagram for a fluid supply system driven by pressurized air through a liquid is shown according to some embodiments of the present invention.

[0013] Figure 4 A fluid flow diagram of a fluid supply system having a working fluid supply pump according to certain embodiments of the present invention is shown.

[0014] Figure 5 A fluid flow diagram of a fluid supply system having a vacuum source according to some embodiments of the present invention is shown.

[0015] FIG. 6A to FIG. 6B A fluid flow diagram of a fluid supply system having a single fluid connection including a quick-connect fluid connector is shown according to certain embodiments of the present invention.

[0016] Figure 7 A fluid flow diagram of a fluid supply system having a single fluid connection including a three-way valve according to some embodiments of the present invention is shown.

[0017] Figure 8 A fluid flow diagram of a fluid supply system having a flow control circuit including independent two-way valve groups according to certain embodiments of the present invention is shown.

[0018] Fig. 9 A fluid flow diagram of a fluid supply system having a flow control circuit including a two-way valve set mounted on a manifold according to certain embodiments of the present invention is shown.

[0019] Fig.10 A fluid flow diagram of a fluid supply system having a flow control circuit including an independent three-way valve set according to certain embodiments of the present invention is shown.

[0020] Fig.11 A fluid flow diagram of a fluid supply system having a flow control circuit including a three-way valve set mounted on a manifold according to certain embodiments of the present invention is shown.

[0021] Fig.12A fluid flow diagram of a fluid supply system having a flow control circuit including a multi-port selector valve according to certain embodiments of the present invention is shown.

[0022] Fig.13 A flow chart is provided of the main functions of a fluid management system of a particle analyzer configured to analyze a sample according to certain embodiments.

[0023] Fig.14 A flow chart of sub-functions within the main functions of a fluid management system of a particle analyzer configured to analyze a sample according to certain embodiments is provided.

[0024] Fig.15 Schematic diagrams of embodiments of a fluid management system of a particle analyzer configured to analyze a sample are provided.

[0025] Fig.16 A flow chart is provided of the main functions of a fluid management system of a particle sorter configured to analyze and sort a sample according to certain embodiments.

[0026] Fig.17 A flow chart of sub-functions within the main functions of a fluid management system of a particle sorter configured to analyze and sort a sample is provided in accordance with certain embodiments.

[0027] Fig.18 Schematic diagrams of embodiments of a fluid management system for a particle sorter configured to analyze and sort samples are provided.

[0028] Fig.19 A flow chart is provided of the main functions of a fluid management system configured as a hybrid system to alternately (a) analyze a sample or (b) analyze and sort a sample, according to certain embodiments.

[0029] Fig. 20 A flow chart of sub-functions within the main functions of a fluid management system configured as a hybrid system to alternately (a) analyze a sample or (b) analyze and sort a sample is provided in accordance with certain embodiments.

[0030] Fig.21 Schematic diagrams of embodiments of fluid management systems configured as hybrid systems that alternately (a) analyze samples or (b) analyze and sort samples are provided.

[0031] Fig. 22 A functional block diagram of an instrument including a flow cytometer according to some embodiments of the present invention is shown.

[0032] Fig.23 A block diagram of a control system according to some embodiments of the present invention is shown.

[0033] FIG. 24A to FIG. 24BA schematic diagram of a particle sorting system according to some embodiments of the present invention is shown.

[0034] Fig.25 An image-enabled particle sorter is depicted in accordance with certain embodiments.

[0035] Fig.26 A block diagram of a computing system according to some embodiments of the invention is shown. DETAILED DESCRIPTION

[0036] The present invention provides a fluid supply system. The fluid supply system of interest includes: a working fluid line; a primary flushing fluid line; a secondary fluid line; a flow control circuit fluidically connected to the distal end of the primary flushing fluid line and the distal end of the secondary fluid line, the flow control circuit configured to selectively allow the primary flushing fluid and the secondary fluid to pass through an outlet; and a single fluid connection configured to fluidically couple to the flow control circuit, the working fluid line, and a fluid assembly of an instrument. Methods of analyzing samples and assembling instruments are also provided.

[0037] Before describing the present invention in more detail, it should be understood that the present invention is not limited to the specific embodiments described, as these embodiments may of course vary. It should also be understood that the terminology used herein is only used to describe specific embodiments and is not intended to be limiting, as the scope of the present invention is limited only by the appended claims.

[0038] When providing a numerical range, it should be understood that unless the context clearly stipulates otherwise, each intermediate value between the upper and lower limits of the range (up to one-tenth of the lower limit unit) and any other stated values ​​or intermediate values ​​in the stated range are included within the scope of the present invention. The upper and lower limits of these smaller ranges can be independently included in the smaller ranges and are also included within the scope of the present invention, except for any specific excluded limits within the range. When the range includes one or two limits, the range excluding one or two of these included limits is also included within the scope of the present invention.

[0039] Certain ranges of values ​​given herein are preceded by the term "about." The term "about" is used herein to provide literal support for the exact number that follows it, as well as for numbers that are close to or approximately the number that follows the term. In determining whether a number is close to or approximately a specifically recited number, a close or approximate unrecited number may be a number that, in the context in which it is presented, provides a substantial equivalence to the specifically recited number.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, only representative illustrative methods and materials are now described.

[0041] All publications and patents cited in this specification are incorporated herein by reference as if each individual publication or patent was specifically and individually indicated to be incorporated herein by reference, and are incorporated herein by reference to disclose and describe the methods and / or materials related to the cited publications. Citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Furthermore, the dates of publication provided may be different from the actual publication dates and may need to be independently confirmed.

[0042] It is noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the claims may be drafted to exclude any optional elements. Thus, this statement is intended to serve as antecedent basis for using exclusive terminology such as "solely," "only," and the like when reciting claim elements or for using a "negative" limitation.

[0043] It will be clear to those skilled in the art after reading this disclosure that each individual embodiment described and illustrated herein has discrete components and features that can be easily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Any described method can be performed in the order of events described or in any other order that is logically possible.

[0044] Although the system and method have been or will be described for the purposes of grammatical fluency and functional explanation, it must be expressly understood that unless otherwise stated or expressly formulated pursuant to 35 U.S.C5, the claims should not be construed as necessarily subject to any limitations in construction as to “means” or “steps” limitations, but should be given the full scope of meaning and equivalents provided by the definitions in the claims pursuant to the doctrine of equivalents, and where a claim is expressly formulated pursuant to the construction of 35 U.S.C5, it should be given the full statutory equivalents provided by the authority enacted in 35 U.S.C5.

[0045] Fluid supply system

[0046] Aspects of the present invention include fluid supply systems. The system of the present invention can be configured to selectively supply fluids, for example, to a fluid assembly of an instrument. By "selectively" supplying fluids, it is meant that the fluid supply system is configured to supply one fluid without supplying other fluids. The fluid that is selectively supplied includes minimal (in some cases zero) traces of other fluids. In other words, the fluid supply system is configured to reduce or eliminate residues between fluids during operation. Reducing or eliminating residues can include: (1) reducing or eliminating the amount of fluid that is retained, and / or (2) reducing or eliminating the number of times residues occur. For example, the system of the present invention can be configured to reduce residues by 80% or more, such as 85% or more, such as 90% or more, such as 95% or more, such as 97% or more, such as 98% or more, such as 99% or more, including 100%. In other words, the fluid supply system can be configured to allow 100% thorough cleaning. The fluid supply system can be configured so that a working fluid (e.g., sheath fluid) and one or more primary flushing fluids or secondary fluids flow through the same fluid path.

[0047] The fluid supply system of the present disclosure can be configured to supply fluid to the fluid component (e.g., flow cell) of the instrument. The fluid supply system can be fluidically coupled to the input end of the fluid component (e.g., flow cell). The fluid supply system can be configured to alternately supply (a) working fluid or (b) one or more of the main flushing fluid and the secondary fluid to the input end of the fluid component. "Alternate supply" means that the fluid supply system is configured in such a way that it can supply (a) working fluid to the input end of the flow cell, and can supply (b) one or more of the main flushing fluid and the secondary fluid to the input end of the flow cell, but can only supply (a) working fluid and (b) one or more of the main flushing fluid and the secondary fluid at a time. For example, the fluid supply system can be configured in such a way that it can supply (a) working fluid to the input end of the fluid component, and it can supply (b) one or more of the main flushing fluid or the secondary fluid to the input end of the fluid component, but in some cases supply (a) working fluid, or in other cases supply (b) one or more of the main flushing fluid or the secondary fluid. The fluid supply subsystem can be configured to alternately fluidically couple (a) a working fluid reservoir or (b) one or more primary flushing fluids and / or secondary fluid reservoirs to the input of a fluid assembly. "Alternately fluidically coupled" means that the fluid supply system is configured in such a way that it is capable of fluidically coupling (a) a working fluid reservoir to the input of a fluid assembly, and is capable of fluidically coupling (b) one or more of the primary flushing fluid and / or secondary fluid reservoirs to the input of a fluid assembly, but can only fluidly couple (a) a working fluid reservoir and (b) one or more of the primary flushing fluid and / or secondary fluid reservoirs to the input of a fluid assembly at a time. In some cases, the fluidic coupling can be automatic / automated so that no user intervention is required. Such automation can be achieved, for example, using a processor-controlled valve.

[0048] Fluid supply system of the present invention includes a working fluid pipeline, which is configured to transport working fluid from the proximal end to the distal end. The working fluid pipeline may include, for example, a tube or a conduit, and may be made of any convenient material. Interested materials include, but are not limited to, polymer materials, such as plastic materials. In some cases, the working fluid pipeline is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), polyethylene (Tygon), fluorinated ethylene-propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), and polyethylene (PE) and the like and their combinations. The working fluid pipeline can have any convenient inner diameter (i.e., the diameter inside the working fluid pipeline), outer diameter (i.e., the diameter outside the working fluid pipeline), wall thickness and length. In some cases, the inner diameter range of the working fluid pipeline is 0.2mm to 10mm, including 1mm to 5mm. In some cases, the outer diameter range of the working fluid pipeline is 0.2mm to 10mm, including 1mm to 5mm. The working fluid pipeline of the present invention also includes a working fluid reservoir connector located at the proximal end, and the working fluid reservoir connector is configured to be fluidically coupled to the working fluid reservoir containing the working fluid." Fluidic coupling" refers to coupling in a manner sufficient to allow fluid to pass through. Any suitable connector can be included. Exemplary connectors include, for example, quick disconnect connectors, threaded connectors, Luer connectors, multi-port connectors, three-clamp connectors, and puncture and seal aseptic connectors. Suitable quick disconnect connectors include, but are not limited to, snap-on (ball lock) connectors, bayonet connectors, threaded connectors, non-locking connectors, single-close connectors, double-close connectors, non-close connectors, dry disconnect connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0049] The working fluid reservoir can be any suitable reservoir or container (e.g., with rigid or flexible walls) for holding the working fluid. The working fluid can vary, but in some cases it is a sheath fluid. The sheath fluid can include, for example, phosphate buffered saline (PBS), HEPES buffered saline, sodium fluoride (NaF), etc., and combinations thereof. The volume range of the working fluid reservoir can be 1L to 100L; for example, the volume range of the container can be 1L to 90L, 1L to 80L, 1L to 70L, 1L to 60L, 1L to 50L, 1L to 40L, 1L to 30L, 1L to 20L or 1L to 10L. In some cases, the working fluid reservoir connector is configured to be detachably connected to the working fluid reservoir. "Detachably connected" means that the connector can be connected to a second connector (e.g., a mating connector) as needed, and then subsequently removed from the second connector at a certain point. In some embodiments, the working fluid reservoir connector provides a sterile connection to the working fluid reservoir. "Sterile" means that the connection is isolated from live bacteria or other microorganisms present in the surrounding environment. If desired, the working fluid reservoir (eg, sheath fluid container) can be cleaned separately, for example, by using an autoclave cycle.

[0050] The present fluid supply system also includes a main flushing fluid line, which is configured to deliver the main flushing fluid from the proximal end to the distal end. The main flushing fluid line may include, for example, a tube or a catheter, and may be made of any convenient material. Interested materials include, but are not limited to, polymer materials, such as plastic materials. In some cases, the working fluid line is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), polyethylene (Tygon), fluorinated ethylene-propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), polyethylene (PE), etc. and combinations thereof. The features of the main flushing fluid line may have any convenient inner diameter (i.e., the diameter inside the main flushing fluid line), outer diameter (i.e., the diameter outside the main flushing fluid line), wall thickness and length. In some cases, the inner diameter range of the main flushing fluid line is 0.2mm to 10mm, including 1mm to 5mm. In some cases, the outer diameter range of the main flushing fluid line is 0.2mm to 10mm, including 1mm to 5mm. The main flushing fluid line of the present invention also includes a main flushing fluid reservoir connector located at the proximal end, which is configured to be fluidically coupled to a main flushing fluid reservoir containing a main flushing fluid. Any suitable connector may be included. Exemplary connectors include, for example, quick disconnect connectors, threaded connectors, Luer connectors, multi-port connectors, tri-clamp connectors, and puncture and seal sterile connectors. Suitable quick disconnect connectors include, but are not limited to, snap-on (ball lock) connectors, bayonet connectors, threaded connectors, non-locking connectors, single-close connectors, double-close connectors, non-close connectors, dry disconnect connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0051] The main flushing fluid reservoir can be any suitable reservoir or container (e.g., having rigid or flexible walls) for accommodating the main flushing fluid. The main flushing fluid can vary. In some cases, the main flushing fluid is an inert fluid. An "inert" fluid refers to a non-reactive fluid. In some embodiments, the main flushing fluid can be mixed with other fluids (e.g., working fluid, secondary fluid) used in the present invention so that when the main flushing fluid is used to flush the fluid supply system, any residual working fluid or secondary fluid will be taken out of the fluid supply system in the main flushing fluid. Exemplary main flushing fluids include, for example, deionized water or saline solutions (phosphate buffered saline, HEPES buffered saline, sodium fluoride solution, etc.). The volume range of the main flushing fluid reservoir can be 1L to 100L; for example, the volume range of the container can be 1L to 90L, 1L to 80L, 1L to 70L, 1L to 60L, 1L to 50L, 1L to 40L, 1L to 30L, 1L to 20L or 1L to 10L. In some cases, the main flush fluid reservoir connector is configured to be removably connected to the main flush fluid reservoir. In some embodiments, the main flush fluid reservoir connector provides a sterile connection to the working fluid reservoir. When necessary, the main flush fluid reservoir can be cleaned separately, such as by using an autoclave sterilization cycle.

[0052] Fluid supply system of the present invention also includes a secondary fluid line, which is configured to transport secondary fluid from the proximal end to the distal end. The secondary fluid line may include, for example, a tube or a conduit, and may be made of any convenient material. Interested materials include, but are not limited to, polymer materials, such as plastic materials. In some cases, the working fluid line is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), polyethylene (Tygon), fluorinated ethylene-propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), polyethylene (PE), etc. and combinations thereof. The secondary fluid line may have any convenient inner diameter (i.e., the diameter inside the secondary fluid line), outer diameter (i.e., the diameter outside the secondary fluid line), wall thickness and length. In some cases, the inner diameter range of the secondary fluid line is 0.2mm to 10mm, including 1mm to 5mm. In some cases, the outer diameter range of the secondary fluid line is 0.2mm to 10mm, including 1mm to 5mm. The secondary fluid line of the present invention also includes a secondary fluid reservoir connector at the proximal end, which is configured to be fluidically coupled to a secondary fluid reservoir containing a secondary fluid. Any suitable connector may be included. Exemplary connectors include, for example, quick disconnect connectors, threaded connectors, Luer connectors, multi-port connectors, tri-clamp connectors, and puncture seal sterile connectors. Suitable quick disconnect connectors include, but are not limited to, snap-on (ball lock) connectors, bayonet connectors, threaded connectors, non-locking connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, dry disconnect connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0053] The secondary fluid reservoir can be any suitable reservoir or container (e.g., with rigid or flexible walls) for accommodating the secondary fluid. The secondary fluid can vary. The secondary fluid can include, for example, a cleaning fluid, a buffer, a calibration solution, a bead solution, etc. In some cases, the secondary fluid is selected from a cleaning fluid and a reagent. When the secondary fluid is a cleaning fluid, any fluid suitable for disinfecting the fluid supply system and / or the fluid assembly / instrument connected thereto can be included. In some embodiments, the cleaning fluid has sterilization and / or antibacterial properties. When the secondary fluid is a reagent, any reagent suitable for use in a flow-type microfluidic environment can be used. The volume range of the secondary fluid reservoir can be 1L to 100L; for example, the volume range of the container can be 1L to 90L, 1L to 80L, 1L to 70L, 1L to 60L, 1L to 50L, 1L to 40L, 1L to 30L, 1L to 20L or 1L to 10L. In some cases, the secondary fluid reservoir connector is configured to be detachably connected to the secondary fluid reservoir. In some embodiments, the secondary fluid reservoir connector provides a sterile connection to the secondary fluid reservoir. If necessary, the secondary fluid reservoir can be cleaned separately, such as by using an autoclave sterilization cycle.

[0054] Embodiments of the fluid supply system include multiple secondary fluid lines and / or secondary fluid reservoirs. In some embodiments, the number of secondary fluid lines in the fluid supply system ranges from 2 to 15, such as 2 to 10, including 2 to 5. In certain versions, the fluid supply system includes 2 or more secondary fluid lines, such as 3 or more secondary fluid lines, such as 4 or more secondary fluid lines, such as 5 or more secondary fluid lines, such as 6 or more secondary fluid lines, such as 7 or more secondary fluid lines, such as 8 or more secondary fluid lines, such as 9 or more secondary fluid lines, including 10 or more secondary fluid lines. In some cases, each secondary fluid line in the fluid supply system is operably connected to a secondary fluid reservoir. The fluids in the secondary fluid reservoirs can be the same or different. In one example including 3 secondary fluid lines and secondary fluid reservoirs, a first secondary fluid reservoir includes a cleaning fluid, a second secondary fluid reservoir includes a reagent, and a third secondary fluid reservoir includes a different reagent.

[0055] The system of the present invention also includes a flow control circuit fluidically connected to the distal end of the main flushing fluid pipeline and the distal end of the secondary fluid pipeline. The flow control circuit of the present invention is configured to selectively allow the main flushing fluid and the secondary fluid to pass through the outlet. In other words, the flow control circuit of the present invention can be configured to allow certain fluids to pass through and prevent other fluids from passing through. In some cases, the flow control circuit is configured to allow a single (i.e., 1) fluid to pass through at a given time. In other cases, the flow control circuit is configured to allow multiple fluids to pass through and prevent other fluids from passing through. The mechanism for achieving selectively allowing fluids may be different. In some versions, the flow control circuit includes a valve group. The number of valves in the group can range from 3 to 10, including 4 to 6. In some cases, the valve group includes 2 or more valves, such as 3 or more valves, such as 4 or more valves, such as 5 or more valves, such as 6 or more valves, such as 7 or more valves, such as 8 or more valves, such as 9 or more valves, and including 10 or more valves. In some versions, the flow control circuit includes a two-way valve group (i.e., a valve with one inlet and one outlet). In some such versions, the flow control circuit includes a valve for each main flushing fluid reservoir and a secondary fluid reservoir. In other embodiments, the flow control circuit includes a three-way valve group. In some such embodiments, the main flushing fluid line is connected to the secondary fluid line via a three-way valve. Additional three-way valves can be added for each additional secondary fluid. In some cases, the valve (e.g., two-way valve, three-way valve) is an independent valve. "Independent" valve means that each valve in the valve group is a separate entity, and they are not installed on the same substrate (e.g., manifold). In other cases, the valve (e.g., two-way valve, three-way valve) is installed on the manifold. In other words, the flow control circuit can include a manifold valve, which includes individual valves. In some embodiments, the distal end of the main flushing fluid line is fluidically coupled to the valve in the valve group that is farthest from the outlet of the flow control circuit. The outlet of the flow control circuit can be defined as the position where the fluid leaves and the valve is closest to a single fluid connection relative to the fluid path, or the position closest to the position. In other words, the inert flush fluid connection is farthest from the flow control loop outlet to achieve maximum flushing of the other paths, thereby minimizing dead volume and fluid carryover between running different fluids.

[0056] In other cases, the flow control circuit includes a multi-port selection valve. The multi-port selection valve discussed herein may include 3 or more ports, such as 4 or more ports, such as 5 or more ports, such as 6 or more ports, such as 7 or more ports, such as 8 or more ports, such as 9 or more ports, and including 10 or more ports. The multi-port selection valve described herein can be configured to selectively allow access between ports. For example, the multi-port selection valve can be connected to a main flushing fluid line and one or more secondary fluid lines, but only allows one type of fluid to pass through at a time. In some embodiments, the multi-port selection valve has a low internal volume, for example, the multi-port selection valve has a volume from 5 mm 3 Up to 1,000mm 3 The internal volume, for example 20mm 3 Up to 100mm 3 , and includes 500mm 3 Up to 1000mm 3 The low internal volume ensures that the entire multi-port selector valve is completely swept with the primary flushing fluid and / or the secondary fluid.

[0057] The fluid supply system of the present invention also includes a single fluid connection, which is configured to be fluidically coupled to the flow control circuit, the working fluid pipeline, and the fluid components of the instrument. A "single" fluid connection means that the fluid from the flow control circuit and the working fluid pipeline passes through the same fluid connection. In other words, there is no other fluid path through which the fluid from the flow control circuit and the working fluid pipeline can pass before being supplied to the fluid components of the instrument. In other words, the single fluid connection is shared between the working fluid reservoir and the flow control circuit, and there is no additional and uncleanable pipe or passage between the working fluid reservoir and the fluid connection. This allows the primary flushing fluid and / or secondary fluid to flush 100% of the passage between the working fluid reservoir and the instrument to achieve very low residuals. The single fluid connection can take any suitable form. In some cases, the single fluid connection is a quick-connect fluid connector. In some such cases, the single fluid connection can be manually changed between the working fluid reservoir and the flow control circuit. Suitable connectors include, but are not limited to, snap-on (ball lock) connectors, bayonet connectors, threaded connectors, non-locking connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, dry disconnect connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors. In other cases, the single fluid connection is a valve, such as a three-way valve. In some embodiments, the three-way valve has a low internal volume, such as a three-way valve having a range from 5 mm to 10 mm. 3 Up to 100mm 3 The internal volume, for example, 15mm 3 Up to 30mm 3, and includes 50mm 3 Up to 80mm 3 The low internal volume ensures that the entire three-way valve is completely swept by the primary flushing fluid and / or the secondary fluid. In some cases where the single fluid connection is a valve, the valve can be configured to be automatically selected, i.e., it can select between the working fluid reservoir and the flow control circuit without manual intervention.

[0058] In some cases, the fluid supply system includes a fluid supply pump operably connected to the flow control circuit and a single fluid connection. The fluid supply pump can be configured to extract fluid from any one of the primary flushing fluid reservoir and the secondary fluid reservoir according to the setting of the flow control circuit. In some cases, the fluid supply pump is a positive displacement pump. As used herein, a "positive displacement pump" refers to a pump that moves fluid by capturing a fixed amount of fluid and forcing (discharging) the captured volume out of the device, wherein such a pump can operate in a series of working cycles, each cycle capturing a certain volume of fluid and mechanically moving the fluid through the pump and into the fluid system. Positive displacement pumps that can be used include, but are not limited to: rotary positive displacement pumps, such as peristaltic pumps, internal gear pumps, screw pumps, shuttle block pumps, flexible vane or sliding vane pumps, circumferential piston pumps, flexible impeller pumps, helical twisted roots pumps or liquid ring pumps; reciprocating positive displacement pumps, such as piston pumps, plunger pumps or diaphragm pumps; and linear positive displacement pumps, such as rope pumps and chain pumps. In certain embodiments, the positive displacement pump comprises a pump selected from the group consisting of a peristaltic pump, a gear pump, and a diaphragm pump. In some cases, the positive displacement pump is a peristaltic pump.

[0059] Figure 1 A block diagram of a fluid supply system according to some embodiments of the present invention is shown. The fluid supply system 100 includes a working fluid reservoir 101, a primary flush fluid reservoir 102, and a secondary fluid reservoir group 103. The primary flush fluid reservoir 102 and the secondary fluid reservoir 103 are both fluidically connected to a flow control circuit 104, which is configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet. The outlet of the flow control circuit 104 is fluidically coupled to a fluid supply pump 105, which draws fluid from one of the primary flush fluid reservoir 104 or the secondary fluid reservoir 103 depending on the setting of the flow control circuit 104. The fluid supply system 100 also includes a single fluid connection 106, which can be fluidically connected to the flow control circuit 104 (via the fluid supply pump 105) and the working fluid reservoir 101. A single fluid connection 106 allows either working fluid from the working fluid reservoir 101 or fluid from the flow control loop 104 to flow to the instrument 107, but not both at the same time.

[0060] Figure 21 shows a fluid flow diagram of a fluid supply system according to some embodiments of the present invention. The fluid supply system 200 includes a working fluid reservoir 201, a primary flushing fluid reservoir 202, and a secondary fluid reservoir group 203. A first secondary fluid reservoir 203a, a second secondary fluid reservoir 203b, and an nth secondary fluid reservoir 203n are shown in the secondary fluid reservoir group 203. The number of secondary fluid reservoirs in the secondary fluid reservoir group 203 can vary, so that n ranges from, for example, 3 to 10. The primary flushing fluid line 208 fluidically couples the primary flushing fluid reservoir 202 to the flow control circuit 204, while the secondary fluid lines 209a, 209b, and 209n fluidically couple the first secondary fluid reservoir 203a, the second secondary fluid reservoir 203b, and the nth secondary fluid reservoir 203n to the flow control circuit 204, respectively. It will be appreciated that there may be additional lines between 209b and 209n depending on the number of secondary fluid reservoirs in the set 203. The outlet of the flow control loop 204 is fluidly coupled to a fluid supply pump 205, which draws fluid from either the primary flush fluid reservoir 204 or one of the secondary fluid reservoirs in the set 203, depending on the configuration of the flow control loop 204. The fluid supply system 200 also includes a single fluid connection 206 that can be fluidly connected to the flow control loop 204 (via the fluid supply pump 205) and the working fluid reservoir 201. The single fluid connection 206 allows either working fluid from the working fluid reservoir 201 or fluid from the flow control loop 204 to flow to the instrument 207 at a given time, but does not allow both to pass through at the same time.

[0061] The system of the present invention can be configured to drive the working fluid to the fluid assembly (e.g., flow cell) of the instrument by any suitable means. An embodiment of the system includes a pressurized air source operably connected to the working fluid reservoir. In such an embodiment, a pressurized air source can be used to apply positive pressure to the working fluid reservoir, thereby driving the working fluid contained therein through the fluid supply system, for example, to the fluid assembly of the instrument. The pressurized air source includes, for example, a compressor and a tank containing pressurized gas. Any suitable compressor can be included. In some embodiments, the compressor is a positive displacement compressor, such as a rotary compressor (e.g., a vane compressor, a screw compressor, a liquid ring compressor, a scroll compressor, a vane compressor) or a reciprocating compressor (e.g., a diaphragm compressor, a double-acting compressor, a single-acting compressor). In other cases, the compressor is a dynamic compressor, such as a centrifugal compressor or an axial compressor. The system can also include a pressure regulator operably connected to the working fluid reservoir and the pressurized air source. Any suitable pressure regulator can be used, for example, a lift valve regulator, a diaphragm chamber regulator, a balanced lift regulator, and a precision regulator can be included. The regulator of interest may be configured to regulate the pressure of pressurized air applied to the working fluid reservoir, thereby regulating the flow rate of the flow stream.

[0062] Figure 3 A fluid flow diagram of a fluid supply system 300 driven by pressurized air through a liquid according to some embodiments of the present invention is shown. The fluid supply system 300 includes a working fluid reservoir 301, a primary flush fluid reservoir 302, a secondary fluid reservoir group 303 (including secondary fluid reservoirs 303a, 303b and 303n), a primary flush fluid line 308, secondary fluid lines 309a-309n, a flow control circuit 304, a fluid supply pump 305, a single fluid connection 306, an instrument 307 and a working fluid line 310. The arrangement of the above elements is as described above with respect to Figure 2 The fluid supply system 300 also includes a pressurized air source 312 and a pressure regulator 311. The pressurized air source 312 is operably connected to the working fluid reservoir 301 and is configured to drive the working fluid therein through the working fluid pipeline 310 and to the instrument 307 through the single fluid connection 306. The pressure regulator 311 is configured to control the amount of pressure applied and the flow rate of the working fluid.

[0063] In other cases, the system includes a working fluid supply pump configured to draw fluid from a working fluid reservoir and supply the working fluid to a fluid component of the instrument (e.g., a flow cell). The working fluid supply pump can be a positive displacement pump. Positive displacement pumps that can be used include, but are not limited to: rotary positive displacement pumps, such as peristaltic pumps, internal gear pumps, screw pumps, shuttle block pumps, flexible vane or sliding vane pumps, circumferential piston pumps, flexible impeller pumps, helical twisted roots pumps, or liquid ring pumps; reciprocating positive displacement pumps, such as piston pumps, plunger pumps, or diaphragm pumps; and linear positive displacement pumps, such as rope pumps and chain pumps. In some embodiments, the positive displacement pump includes a pump selected from the group consisting of a peristaltic pump, a gear pump, and a diaphragm pump. In some cases, the positive displacement pump is a peristaltic pump.

[0064] Figure 4 A fluid flow diagram of a fluid supply system 400 with a working fluid supply pump according to some embodiments of the present invention is shown. The fluid supply system 400 includes a working fluid reservoir 401, a primary flushing fluid reservoir 402, a secondary fluid reservoir group 403 (including secondary fluid reservoirs 403a, 403b and 403n), a primary flushing fluid line 408, secondary fluid lines 409a-409n, a flow control circuit 404, a fluid supply pump 405, a single fluid connection 406, an instrument 407 and a working fluid line 410. The arrangement of the above elements is as described above with respect to Figure 2 The fluid supply system 400 further includes a working fluid supply pump 411 located between the instrument 407 and the single fluid connection 406 . The working fluid supply pump 411 is configured to draw working fluid from the working fluid reservoir 401 and provide it to the instrument 407 .

[0065] In other cases, the fluid supply system includes a vacuum source operably connected to the fluid assembly of the instrument. In some cases, the vacuum source is a pump. The vacuum source can be located on the opposite side of the fluid assembly of the instrument (e.g., flow cell) relative to a single fluid connection, so that the vacuum generated by the vacuum source draws the working fluid out of the working fluid reservoir through the single fluid connection and through the fluid assembly of the instrument (e.g., flow cell). Any pump suitable for generating a vacuum capable of driving a fluid from the working fluid reservoir can be used, including but not limited to the pump described herein.

[0066] Figure 5A fluid flow diagram of a fluid supply system 500 with a vacuum source according to some embodiments of the present invention is shown. The fluid supply system 500 includes a working fluid reservoir 501, a primary flushing fluid reservoir 502, a secondary fluid reservoir group 503 (including secondary fluid reservoirs 503a, 503b and 503n), a primary flushing fluid line 508, secondary fluid lines 509a-509n, a flow control circuit 504, a fluid supply pump 505, a single fluid connection 506, an instrument 507 and a working fluid line 510. The arrangement of the above elements is as described above with respect to Figure 2 The fluid supply system 500 also includes a vacuum source 511 located on an opposite side of the instrument 507 relative to the single fluid connection, such that the vacuum created by the vacuum source 511 draws the working fluid from the working fluid reservoir 501 through the single fluid connection 506 and through the instrument 507 .

[0067] FIG. 6A to FIG. 6B A fluid flow diagram of a fluid supply system 600 having a single fluid connection including a quick connect fluid connector according to some embodiments of the present invention is shown. The fluid supply system 600 includes a working fluid reservoir 601, a primary flush fluid reservoir 602, a secondary fluid reservoir set 603 (including secondary fluid reservoirs 603a, 603b, and 603n), a primary flush fluid line 608, secondary fluid lines 609a-609n, a flow control circuit 604, a fluid supply pump 605, an instrument 607, and a working fluid line 610. The arrangement of the above elements is as described above with respect to Figure 2 The fluid supply system 600 also includes a single fluid connection consisting of a quick connect fluid connector 606 that can be fluidly coupled to either the working fluid reservoir 601 or the flow control circuit 604 at a given time. Fig. 6A The quick connect fluid connector 606 is shown fluidly coupled to the working fluid reservoir 601, while Figure 6B A quick connect fluid connector 606 is shown fluidly coupled to the flow control circuit 604 .

[0068] Figure 7 A fluid flow diagram of a fluid supply system 700 having a single fluid connection including a three-way valve according to some embodiments of the present invention is shown. The fluid supply system 700 includes a working fluid reservoir 701, a primary flush fluid reservoir 702, a secondary fluid reservoir group 703 (including secondary fluid reservoirs 703a, 703b, and 703n), a primary flush fluid line 708, secondary fluid lines 709a-709n, a flow control circuit 704, a fluid supply pump 705, an instrument 707, and a working fluid line 710. The arrangement of the above elements is as described above with respect to Figure 2The fluid supply system 700 also includes a single fluid connection 706 consisting of a three-way valve 706. The three-way valve 706 has two inlets (one inlet each fluidically coupled to the working fluid reservoir 701 and the flow control loop 704) and an outlet fluidically coupled to the instrument 707. Depending on the setting of the three-way valve 706, fluid from the working fluid reservoir 701 or the flow control loop 704 can be provided to the instrument 707. In some cases, the three-way valve 706 can be automatically controlled, such as by a processor.

[0069] Figure 8 A fluid flow diagram of a fluid supply system 800 having a flow control circuit including an independent two-way valve group according to some embodiments of the present invention is shown. The fluid supply system 800 includes a working fluid reservoir 801, a primary flushing fluid reservoir 802, a secondary fluid reservoir group 803 (including secondary fluid reservoirs 803a, 803b and 803n), a primary flushing fluid line 808, secondary fluid lines 809a-809n, a fluid supply pump 805, a single fluid connection 806, an instrument 807 and a working fluid line 810. The arrangement of the above elements is as described above with respect to Figure 2 As described. The fluid supply system 800 also includes a flow control circuit consisting of a set of independent two-way valves 804a-804d. Each of the two-way valves 804a-804d can be individually actuated so that when the valve is open, fluid can be supplied from the corresponding reservoir, and when the valve is closed, the supply of fluid is prevented. The opening and closing of the valve can be automatically controlled, for example, by a processor in some cases. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines (for example, as described above) can be included in the fluid supply system 800. If this is the case, an additional two-way valve can be included for each additional secondary fluid reservoir and secondary fluid line. As Figure 8 As shown, the main flush fluid line 808 is fluidly coupled to the valve 804a in the valve group that is farthest from the outlet of the flow control circuit (i.e., the location where valves 804a-804d are fluidly connected to the fluid supply pump 805). This ensures that the fluid lines in the flow control circuit are completely purged with the main flush fluid.

[0070] Fig. 9 A fluid flow diagram of a fluid supply system 900 having a flow control circuit including a co-mounted two-way valve group according to some embodiments of the present invention is shown. The fluid supply system 900 includes a working fluid reservoir 901, a primary flush fluid reservoir 902, a secondary fluid reservoir group 903 (including secondary fluid reservoirs 903a, 903b, and 903n), a primary flush fluid line 908, secondary fluid lines 909a-909n, a fluid supply pump 905, a single fluid connection 906, an instrument 907, and a working fluid line 910. The arrangement of the above elements is as described above with respect to Figure 2 As described. The fluid supply system 900 also includes a flow control circuit, which includes a two-way valve group 904a-904d mounted on the manifold 904. Each of the two-way valves 904a-904d can be individually actuated so that fluid can be provided from the corresponding reservoir when the valve is open, and the supply of fluid is blocked when the valve is closed. In some cases, the opening and closing of the valve can be automatically controlled, such as by a processor. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines (for example, as described above) may be included in the fluid supply system 900. If this is the case, an additional two-way valve may be included on the manifold 904 for each additional secondary fluid reservoir and secondary fluid line. As Fig. 9 As shown, the main flush fluid line 908 is fluidly coupled to the valve 904a in the manifold 904 that is farthest from the outlet of the flow control circuit (i.e., where valves 904a-904d are fluidly connected to the fluid supply pump 905). This ensures that the fluid lines in the flow control circuit are completely purged with the main flush fluid.

[0071] Fig.10 A fluid flow diagram of a fluid supply system 1000 having a flow control circuit including an independent three-way valve group according to some embodiments of the present invention is shown. The fluid supply system 1000 includes a working fluid reservoir 1001, a primary flushing fluid reservoir 1002, a secondary fluid reservoir group 1003 (including secondary fluid reservoirs 1003a, 1003b and 1003n), a primary flushing fluid line 1008, secondary fluid lines 1009a-1009n, a fluid supply pump 1005, a single fluid connection 1006, an instrument 1007 and a working fluid line 1010. The arrangement of the above elements is as described above with respect to Figure 2The fluid supply system 1000 also includes a flow control circuit composed of independent three-way valves 1004a, 1004b and 1004c. The three-way valve 1004a has an inlet fluidically coupled to the main flushing fluid reservoir 1002 via the main flushing fluid pipeline 1008, and another inlet fluidically coupled to the secondary fluid reservoir 1003a. The three-way valve 1004b has an inlet fluidically coupled to the outlet of the three-way valve 1004a, and another inlet fluidically coupled to the secondary fluid reservoir 1003b via the secondary fluid pipeline 1009a. The three-way valve 1004c has an inlet fluidically coupled to the outlet of the three-way valve 1004b, and another inlet fluidically coupled to the secondary fluid reservoir 1003n via the secondary fluid pipeline 1009n. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid pipelines (e.g., as described above) may be included in the fluid supply system 1000. If this is the case, an additional three-way valve may be included for each additional secondary fluid reservoir and secondary fluid line. Each of the three-way valves 1004a-1004c may be individually actuated so that fluid may be provided from the corresponding reservoir when the valve is open, and fluid is prevented from being provided when the valve is closed. Fig.10 As shown, the main flush fluid line 1008 is fluidly coupled to the valve 1004a farthest from the outlet of the flow control circuit (i.e., where valves 1004a-1004c are fluidly connected to the fluid supply pump 1005). This ensures that the fluid lines in the flow control circuit are completely purged with the main flush fluid.

[0072] Fig.11 A fluid flow diagram of a fluid supply system 1100 having a flow control circuit including a three-way valve group mounted on a manifold according to some embodiments of the present invention is shown. The fluid supply system 1100 includes a working fluid reservoir 1101, a primary flush fluid reservoir 1102, a secondary fluid reservoir group 1103 (including secondary fluid reservoirs 1103a, 1103b, and 1103n), a primary flush fluid line 1108, secondary fluid lines 1109a-1109n, a fluid supply pump 1105, a single fluid connection 1106, an instrument 1107, and a working fluid line 1110. The arrangement of the above elements is as described above with respect to Figure 2The fluid supply system 1100 also includes a flow control circuit including three-way valves 1104a, 1104b, and 1104c mounted on the manifold 1104. The three-way valve 1104a has one inlet fluidically coupled to the primary flushing fluid reservoir 1102 via the primary flushing fluid line 1108, and another inlet fluidically coupled to the secondary fluid reservoir 1103a via the secondary fluid line 1109a. The three-way valve 1104b has one inlet fluidically coupled to the outlet of the three-way valve 1104a, and another inlet fluidically coupled to the secondary fluid reservoir 1103b via the secondary fluid line 1109b. The three-way valve 1104c has one inlet fluidically coupled to the outlet of the three-way valve 1104b, and another inlet fluidically coupled to the secondary fluid reservoir 1103n via the secondary fluid line 1109n. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines (e.g., as described above) may be included in the fluid supply system 1100. If this is the case, an additional three-way valve may be included on the manifold for each additional secondary fluid reservoir and secondary fluid line. Each of the three-way valves 1104a-1104c may be individually actuated so that fluid may be supplied from the corresponding reservoir when the valve is open, and fluid may be blocked from being supplied when the valve is closed. Fig.11 As shown, the main flush fluid line 1108 is fluidly coupled to the valve 1104a farthest from the outlet of the flow control circuit (i.e., where valves 1104a-1104c are fluidly connected to the fluid supply pump 1105). This ensures that the fluid lines in the flow control circuit are completely purged with the main flush fluid.

[0073] Fig.12 A fluid flow diagram of a fluid supply system 1200 having a flow control circuit including a multi-port selector valve according to some embodiments of the present invention is shown. The fluid supply system 1200 includes a working fluid reservoir 1201, a primary flushing fluid reservoir 1202, a secondary fluid reservoir group 1203 (including secondary fluid reservoirs 1203a, 1203b, and 1203n), a primary flushing fluid line 1208, secondary fluid lines 1209a-1209n, a fluid supply pump 1205, a single fluid connection 1206, an instrument 1207, and a working fluid line 1210. The arrangement of the above elements is as described above with respect to Figure 2The fluid supply system 1200 also includes a multi-port selector valve 1204, which is fluidically connected to the primary flushing fluid reservoir 1202 via the primary flushing fluid line 1208, to the secondary fluid reservoir 1203a via the secondary fluid line 1209a, to the secondary fluid reservoir 1203b via the secondary fluid line 1209b, to the secondary fluid reservoir 1203n via the secondary fluid line 1203n, and to the fluid supply pump 1205. The multi-port selector valve 1204 is configured to selectively route fluid from one of these fluidly connected reservoirs at a given time. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines (e.g., as described above) may be included in the fluid supply system 1200. If this is the case, these reservoirs and secondary fluid lines may also be fluidically connected to the multi-port selector valve 1204.

[0074] Fluid Management System

[0075] Aspects of the present invention also include a fluid management system, which includes the fluid supply system of the present invention. The term "fluid management system" refers to the entirety of the fluid components associated with an instrument (e.g., a flow cytometer). The fluid management system of the present disclosure can be configured to perform one or more functions, including but not limited to supplying a working fluid (e.g., sheath fluid) and a sample fluid to a fluid component (e.g., a flow cell) at a controlled ratio; supplying a primary flush fluid to a fluid component; supplying one or more secondary fluids (e.g., for cleaning or as a reagent) to a fluid component; using a working fluid to perform hydrodynamic focusing of a sample fluid for sample interrogation through a cuvette; collecting analyzed samples as waste; collecting waste generated by sample sorting; and managing and storing all system waste; etc. The fluid management system may include: a flow cell, the flow cell including an input end and an output end; a cuvette, the cuvette including an input end coupled to an output end of the flow cell and also including an output end; a sample input line for fluidically coupling a sample fluid source to the input end of the flow cell; and any fluid supply system described herein. Exemplary fluid management systems that may be adapted to include the subject fluid supply systems include those described in U.S. Patent Application Publication No. 2022 / 0341838, the disclosure of which is incorporated herein by reference.

[0076] Fig.13A flow chart of the main functions of a fluid management system configured to analyze a sample according to certain embodiments is provided. The main functions are: (1301) supplying a working fluid (e.g., sheath fluid) and a sample fluid to a flow cell in controlled proportions; (1302) supplying any number of primary flush fluids and secondary fluids (e.g., for cleaning); (1303) performing hydrodynamic focusing of a sample using the working fluid for sample interrogation through a cuvette; (1304) collecting analyzed samples as waste; and (1305) managing and storing all system wastes (waste from particle analysis 1304 and secondary system waste 1306). Dashed lines indicate that (a) the working fluid and the sample fluid can be supplied to the flow cell, or (b) any number of secondary system fluids can be supplied to the flow cell.

[0077] In some embodiments, the fluid management system is configured to alternately (a) analyze a sample (e.g., particles in a sample fluid) or (b) analyze and sort a sample (e.g., particles in a sample fluid). In such embodiments, the fluid management system may include all components required for (a) analyzing a sample or (b) analyzing and sorting a sample, but may only perform one of (a) and (b) at a time. In some embodiments, the fluid management system is configured such that it is capable of (a) analyzing a sample and is capable of (b) analyzing and sorting a sample, but performs (a) analyzing a sample in some cases or performs (b) analyzing and sorting a sample in other cases.

[0078] The fluid management aspects of the fluid management system can be based on the fluid circuit principle. For example, the basic principle of the working fluid supply can be based on the fluid circuit principle, in which it is assumed that the pressure drop on the closed fluid path is equal to the product of the liquid flow rate and the fluid resistance. The working fluid (e.g., sheath fluid) and the sample path can be modeled as two resistors in parallel, combined to pass through the flow cell, the cuvette and the waste path, which are the third fluid resistor. Similarly, according to certain embodiments, waste management using a single waste pump is based on the fluid circuit principle. Each individual waste liquid suction source can be modeled as a fluid reservoir under atmospheric pressure, and each connection (e.g., fluid pipeline) from these waste liquid sources to the waste liquid management system can be modeled as a fluid resistor. Given the supply vacuum of the waste liquid pump, the fluid resistance of each connection can be controlled to balance the waste liquid suction required for each waste liquid source, as described in further detail below.

[0079] According to certain embodiments, the waste liquid of the flow-type particle analyzer of the present disclosure can be managed by a fluid moving device. In some cases, the waste liquid (e.g., the combined waste liquid) of the fluid management system is managed by a single fluid moving device (e.g., a single waste liquid pump). The combined waste liquid can be managed by a single fluid moving device (e.g., a single waste liquid pump) by properly balancing the fluid resistance of all different waste liquid sources from the system (such as, for example, analyzer waste liquid, sorting waste liquid and secondary system waste liquid). The waste liquid source may include, for example, an analyzer waste liquid collection subsystem, a sorter waste liquid collection subsystem and one or more system waste liquid sources. The connection (e.g., a fluid pipeline) from the waste liquid source can act as a plurality of fluid resistors in parallel with a single pressure source (including a fluid moving device, e.g., a waste liquid vacuum pump). As long as the fluid moving device (e.g., a waste liquid vacuum pump) has sufficient capacity, the resistance can be adjusted so that each waste liquid source has an appropriate vacuum. For example, the resistance of a flow path (e.g., a fluid line) that fluidically couples a waste source of a system to a fluid moving device can have known resistances that are balanced to allow each waste source to have an appropriate vacuum to extract waste. In some cases, the resistance of a first fluid line coupled to a first waste source among a plurality of fluid lines that couple multiple waste sources to a fluid moving device can be adjusted or set to provide an appropriate vacuum for the first waste source. The resistance of a second fluid line coupled to a second waste source among a plurality of fluid lines that couple multiple waste sources to a fluid moving device can be adjusted or set to provide an appropriate vacuum for the second waste source. The resistance of additional fluid lines in the system can also have individual resistances that are adjusted or set to provide an appropriate vacuum for each respective waste source. The resistance of the fluid lines from each waste source can be the same or different. Each fluid line that fluidically couples a waste source to a fluid moving device can have a specific known resistance so that each waste source has an appropriate vacuum. For example, the resistance of the fluid lines coupling one or more of the analyzer waste collection subsystem, the sorter waste collection subsystem, and one or more system waste sources to the fluid moving device can be balanced to allow for an appropriate vacuum for each waste source and control waste aspiration of the individual fluid moving devices. In some cases, the resistance of multiple fluid lines coupling multiple waste sources to the fluid moving device can be individually adjusted to provide a desired fluid flow rate from each respective waste source. For example, the resistance of a first fluid line coupled to a first waste source among the multiple fluid lines coupling the multiple waste sources to the fluid moving device can be adjusted or set to provide a desired first flow rate of waste from the first waste source. The resistance of a second fluid line coupled to a second waste source among the multiple fluid lines coupling the multiple waste sources to the fluid moving device can be adjusted or set to provide a desired second flow rate of waste from the second waste source.The resistance of the additional fluid lines in the system can also have individual resistances, which are adjusted or set to provide the required flow rate of the waste liquid from each corresponding waste liquid source. The resistance of the fluid lines from each waste liquid source can be the same or different. Each fluid line that couples the waste liquid source to the fluid moving device can have a specific known resistance based on the required fluid flow rate from each waste liquid source. In some cases, the resistance of the flow path (e.g., fluid line) in the fluid management system is fixed. In some cases, the resistance of the flow path (e.g., fluid line) in the fluid management system is variable. In some cases, the resistance of one or more flow paths in the system can be changed independently, wherein, for example, the resistance of each flow path can be changed independently. In some cases, the system includes one or more variable resistors, which can adjust the resistance of the flow path by, for example, adjusting the size of the flow path (e.g., the diameter or length of the fluid line). In some cases, the variable resistor includes a valve.

[0080] The fluid management system of the present disclosure may also include one or more waste collection subsystems. In some cases, the fluid management system includes: an analyzer waste collection subsystem configured to fluidically couple the output of the cuvette to the waste management subsystem; a sorter waste collection subsystem configured to fluidically couple the output of the sorting block coupled to the output of the cuvette to the waste management subsystem; or a combination thereof (e.g., in a hybrid system, where a flow-type particle analyzer can (a) analyze a sample, or (b) analyze and sort a sample).

[0081] The waste management subsystem may be configured to receive and combine waste from: (a) one or more system waste sources and (b) an analyzer waste collection subsystem, a sorter waste collection subsystem, or a combination thereof to produce combined waste. Fluid from a system waste source may include waste generated and / or collected from any portion of a clean flow-type particle analyzer. Fluid from the analyzer waste collection subsystem may include analyzed waste from the output end of a cuvette, which may include, for example, analyzed sample fluid and working fluid that have flowed through a cuvette and been optically interrogated. Fluid from the sorter waste collection subsystem may include waste received and collected from the output end of a sorting block (e.g., the output end of a sorting nozzle). Waste may include analyzed working fluid and sample fluid (e.g., including particles of no interest) that are not transferred to a sample collection container in the form of droplets by one or more deflection plates of the sorting block. In certain embodiments, the waste management subsystem is configured to receive and merge waste from: (a) one or more system waste sources and (b) an analyzer waste collection subsystem to produce a merged waste. In certain embodiments, the waste management subsystem is configured to receive and merge waste from: (a) one or more system waste sources and (b) a sorter waste collection subsystem to produce a merged waste. In certain embodiments, the waste management subsystem is configured to receive and merge waste from: (a) one or more system waste sources and (b) an analyzer waste collection subsystem and a sorter waste collection subsystem. In certain embodiments, the waste management subsystem is configured to receive and merge waste from: (a) one or more system waste sources and (b) an analyzer waste collection subsystem or a sorter waste collection subsystem.

[0082] The waste management subsystem can be fluidly coupled to (a) one or more system waste sources and (b) an analyzer waste collection subsystem, a sorter waste collection subsystem, or a combination thereof. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste sources and (b) an analyzer waste collection subsystem. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste sources and (b) a sorter waste collection subsystem. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste sources and (b) an analyzer waste collection subsystem and a sorter waste collection subsystem.

[0083] The waste management subsystem may include an output, such as a single output, for combining waste liquids. In some cases, the output of the waste management subsystem is fluidically coupled to a waste container. A fluid line, such as a tube or conduit, may fluidically couple the output of the waste management subsystem to the waste container. In some cases, the output is fluidically coupled to the waste container via a single fluid line. Combined waste liquid may flow from the output and into a waste container, such as for storage. The waste container may be fluidically coupled (e.g., via a fluid line (e.g., a single fluid line)) to a fluid moving device configured to transfer the combined waste liquid from the output of the waste management subsystem to the waste container. In some cases, the waste container may be detachable from the fluid line coupling it to the fluid management system so that, for example, the waste container may be emptied and cleaned.

[0084] If desired, the fluid management system may include a fluid moving device configured to transfer (e.g., mechanically transfer) the combined waste liquid to a waste container. In some cases, the combined waste liquid is managed by a single fluid moving device (e.g., a single pump). In these embodiments, since there is only a single fluid moving device, the fluid management system does not include any other fluid moving device that controls or regulates the flow of the combined waste liquid to the waste container. In some embodiments, a single fluid line may couple the output end of the waste management subsystem to the waste container. In some embodiments, a single fluid line may fluidically couple the output end of the waste management system to a single fluid moving device, which in turn is fluidically coupled to the waste container via a single fluid line. In some cases, the combined waste liquid is transferred from the output end of the waste management subsystem to the waste container via a single fluid moving device (e.g., a single pump). In some cases, the fluid moving device is a vacuum source that draws the combined waste liquid from the output end of the fluid management subsystem to the waste container. In some cases, the fluid moving device transfers waste liquid from each subsystem or waste liquid source (e.g., analyzer waste liquid collection subsystem, sorter waste liquid collection subsystem, system waste liquid source) to the waste liquid management subsystem, for example, to be combined, and then the combined waste liquid is transferred to the waste liquid container. In some cases, the waste liquid from each subsystem or waste liquid source fluidically coupled to the waste liquid management subsystem is managed by a single fluid moving device. In some cases, the fluid moving device is a vacuum source that draws waste liquid from each subsystem or waste liquid source to the waste liquid management subsystem. The fluid moving device can be fluidically coupled to the output end of the waste liquid management subsystem and the waste liquid container. The fluid moving device can be any fluid moving device as described herein. In some cases, the fluid moving device is a vacuum application device. In some cases, the fluid moving device includes a pump, such as a vacuum pump. In some cases, the fluid moving device is a single pump. In some cases, the pump includes a positive displacement vacuum pump. In some cases, the positive displacement vacuum pump includes a pump selected from the group consisting of a diaphragm pump, a gear pump, and a peristaltic pump. In some cases, the positive displacement vacuum pump is a diaphragm pump.

[0085] In some embodiments, the flow-type particle analyzer includes a waste liquid container. The waste liquid container can be any suitable reservoir or container (e.g., having rigid or flexible walls) for storing waste liquid. In some cases, the output end of the waste liquid management subsystem is fluidically coupled to the waste liquid container. In some cases, the waste liquid container is fluidically coupled to a fluid moving device, which is configured to transfer the combined waste liquid from the output end of the waste liquid management subsystem to the waste liquid container. A fluid pipeline (e.g., a tube or a conduit) can fluidically couple the output end of the waste liquid management subsystem to the waste liquid container. In some cases, the output end is fluidically coupled to the waste liquid container through a single fluid pipeline. The combined waste liquid can flow out of the output end and flow into the waste liquid container, for example for storage. The waste liquid container can be fluidically coupled to a fluid moving device, for example, via a fluid pipeline (e.g., a single fluid pipeline), which is configured to transfer the combined waste liquid from the output end of the waste liquid management subsystem to the waste liquid container. In some cases, the waste liquid container can be detached from the fluid pipeline connecting it to the fluid management system so that, for example, the waste liquid container can be emptied and cleaned. The waste container can be configured, for example, to have a suitable volume to hold and store all system fluids. The volume range of the waste container can be 1L to 100L; for example, the volume range of the container can be 1L to 90L, 1L to 80L, 1L to 70L, 1L to 60L, 1L to 50L, 1L to 40L, 1L to 30L, 1L to 20L, or 1L to 10L.

[0086] The fluid management system may include one or more system waste sources fluidically coupled to the waste management subsystem. The one or more system waste sources may generate and / or collect waste from cleaning any area or location of the flow-type particle analyzer. The one or more system waste sources may be configured to transfer or deliver system waste to the waste management subsystem. In some cases, waste from one or more system waste sources is sucked into the waste management subsystem by a vacuum generated by a fluid moving device. The one or more waste sources may be fluidically coupled to the waste management subsystem with a known fluid resistance. In some cases, one or more fluid lines (e.g., each of the one or more fluid lines) coupling the one or more system waste sources to the waste management subsystem include a resistor. The resistor may provide a known fluid resistance in one or more fluid lines coupling the one or more system waste sources to the waste management subsystem. Suitable resistors include, but are not limited to, a section of pipe, an orifice, etc. having a known length and inner diameter.

[0087] The fluid management system of the present disclosure may also include a working fluid pressure feedback control subsystem, which is configured to control or regulate the working fluid pressure in the system (e.g., in a working fluid reservoir). The working fluid pressure may be controlled via the regulated air pressure in the working fluid reservoir. The pressurized working fluid may flow from the working fluid source to the flow pool through the fluid supply system. The working fluid pressure may be controlled based on various parameters, including, for example, the fluid level height measured in the working fluid reservoir and the air pressure in the working fluid reservoir. Other parameters may include, but are not limited to, the working fluid flow rate.

[0088] The working fluid pressure feedback control subsystem may include a controller configured to maintain a controlled variable (e.g., fluid pressure) at a desired set point. The controller may be any suitable control system. In some cases, the controller includes a closed-loop control system, such as a feedback controller. The system may include any suitable sensor for sensing one or more parameters of the feedback signal. In some cases, the controller is configured to receive a signal representing a measured working fluid reservoir gas pressure. In some cases, the controller is configured to receive a signal representing a measured liquid level in the working fluid reservoir. In some cases, the controller is configured to receive a signal representing a measured working fluid reservoir gas pressure and a signal representing a measured liquid level in the working fluid reservoir. In some cases, the controller is configured to determine whether the working fluid pressure deviates from the set point based on the received signal representing the measured working fluid reservoir gas pressure and / or the signal representing the measured liquid level in the working fluid reservoir. In some cases, the controller is configured to send a working fluid pressure control signal to the working fluid reservoir gas pressure regulator when the working fluid pressure deviates from the set point, for example, to adjust the working fluid reservoir gas pressure (and thereby adjust the working fluid pressure and the working fluid flow rate). The working fluid pressure control signal can cause the working fluid reservoir gas pressure regulator to adjust the gas pressure in the working fluid reservoir so that the working fluid pressure matches the desired set point. In some cases, the controller is configured to send a working fluid pressure control signal to a controller of a sample fluid pressure feedback control system. Any suitable feedback control mechanism can be used. The feedback controller of interest can use control parameters such as proportional gain, integral term and / or differential term. The control parameter can be applied to an error signal (e.g., the difference between the set point and the feedback signal) to calculate the input provided to the controlled system or process. In some embodiments, the controller is a proportional integral derivative (PID) controller.

[0089] In some cases, the working fluid pressure feedback control subsystem includes a working fluid reservoir gas pressure regulator, which is configured to control the air pressure in the working fluid reservoir. The working fluid reservoir gas pressure regulator can be configured to receive a working fluid pressure control signal from a controller and adjust the air pressure in the working fluid reservoir (and thereby adjust the fluid pressure). The working fluid reservoir gas pressure regulator can adjust the air pressure in the working fluid reservoir so that the working fluid pressure matches the desired set point. The working fluid reservoir gas pressure regulator can be gaseously coupled to a pressurized air source. In some cases, the working fluid reservoir gas pressure regulator is gaseously coupled to the working fluid reservoir. In some cases, the inlet of the working fluid reservoir gas pressure regulator is gaseously coupled to the pressurized air source. In some cases, the outlet of the working fluid reservoir gas pressure regulator is gaseously coupled to the working fluid reservoir. Any suitable gas pressure regulator can be used. The pressure regulator of interest may include, for example, a device that controls (e.g., regulates) the supply (or inlet) pressure to a desired outlet pressure and works to maintain the outlet pressure in the event of an inlet pressure fluctuation. In some cases, the working fluid reservoir gas pressure regulator is an electro-pneumatic pressure regulator.

[0090] In some cases, the working fluid pressure feedback control subsystem includes a fluid level measuring device configured to measure the fluid level height in the working fluid reservoir. The fluid level measuring device can be operably coupled to the working fluid reservoir. According to the embodiments described herein, the output signal of the fluid level measuring device (e.g., representing the measured fluid level) can be sent to the controller of the working fluid pressure feedback control subsystem. In some cases, the fluid level measuring device is a point level measurement sensor configured to mark a single discrete liquid height. In some cases, the fluid level measuring device is a continuous level sensor configured to measure the fluid level within a certain range. Suitable fluid level measuring devices include, but are not limited to, glass level gauges, float switches, ultrasonic sensors, capacitive level sensors, hydrostatic devices (e.g., bubblers, displacers, differential pressure transmitters), strain gauges, magnetic level gauges, magnetostrictive level transmitters, laser level transmitters, radar level sensors, guided radar sensors, tuning forks, etc.

[0091] The fluid management system of the present disclosure may also include a sample fluid pressure feedback control subsystem configured to control or regulate the sample fluid pressure in the system (e.g., in a sample fluid source). The sample fluid pressure may be controlled via regulated air pressure in the sample fluid source. The pressurized sample fluid may be supplied directly to the flow cell from the sample fluid source. The fluid pressure in the sample fluid source may be controlled based on a variety of parameters, including, for example, air pressure measured in the sample fluid source and a working fluid pressure control signal. Other parameters may include, but are not limited to, sample level height and sample fluid flow rate.

[0092] The sample fluid pressure feedback control subsystem may include a controller configured to maintain a controlled variable (e.g., fluid pressure) at a desired set point. The controller may be any suitable control system. In some cases, the controller includes a closed-loop control system, such as a feedback controller. The system may include any suitable sensor for sensing one or more parameters of the feedback signal. The sample fluid pressure set point may be the sheath fluid pressure + desired flow rate. In this case, the sample fluid pressure will always be greater than the working fluid (e.g., sheath fluid) pressure of the positive flow through the system. This allows dynamic adjustment of the working fluid without changing the effective pressure drop of the sample fluid. The systems described herein may provide control of the sample pressure as a differential above the working fluid (e.g., sheath fluid) pressure. This allows the working fluid pressure to be varied while keeping the sample flow rate constant (P_sample (sample fluid pressure) = P_sheath (sheath fluid pressure) + P_diff (differential pressure). In some embodiments, the controller is configured to receive a signal representing a measured sample fluid source gas pressure. In some embodiments, the controller is configured to receive a working fluid pressure control signal, such as a controller from a working fluid pressure feedback control subsystem. In some embodiments, the controller is configured to receive a signal representing a measured sample fluid source gas pressure and a working fluid pressure control signal. In some cases, the controller is configured to determine whether the sample fluid pressure has deviated from a set point based on the received signal representing the measured sample fluid source gas pressure and / or the working fluid pressure control signal. point. The controller can be configured to send a sample fluid pressure control signal to the sample fluid source gas pressure regulator in the event that the sample fluid pressure has deviated from the set point, for example, to adjust the sample fluid source gas pressure (and thereby adjust the sample fluid pressure and sample fluid flow rate). The sample fluid pressure control signal can cause the sample fluid source gas pressure regulator to adjust the gas pressure in the sample fluid source so that the sample fluid pressure matches the desired set point. Any suitable feedback control mechanism can be used. The feedback controller of interest can use control parameters, such as proportional gain, integral terms and / or differential terms. The control parameters can be applied to an error signal (e.g., the difference between the set point and the feedback signal) to calculate an input provided to the controlled system or process. In some embodiments, the controller is a PID controller.

[0093] In some cases, the sample fluid pressure feedback control subsystem includes a sample fluid source gas pressure regulator configured to control the gas pressure in the sample fluid source. The sample fluid source gas pressure regulator can be configured to receive a sample fluid pressure control signal from a controller and adjust the gas pressure (and thereby adjust the fluid pressure) in the sample fluid source. The sample fluid source gas pressure regulator can adjust the gas pressure in the sample fluid source so that the sample fluid pressure matches the desired set point. The sample fluid source gas pressure regulator can be gaseously coupled to a pressurized air source. In some cases, the sample fluid source gas pressure regulator is gaseously coupled to the sample fluid source. In some cases, the inlet of the sample fluid source gas pressure regulator is gaseously coupled to the pressurized air source. In some cases, the outlet of the sample fluid source gas pressure regulator is gaseously coupled to the sample fluid source. Any suitable gas pressure regulator can be used. The pressure regulator of interest can include, for example, a device that controls (e.g., regulates) a supply (or inlet) pressure to a desired outlet pressure and maintains the outlet pressure despite fluctuations in the inlet pressure. In some cases, the sample fluid source gas pressure regulator is an electro-pneumatic pressure regulator.

[0094] In some embodiments, the flow-type particle analyzer includes a sample fluid source. The sample fluid source can be any suitable reservoir or container (e.g., having rigid or flexible walls) for containing the sample fluid. In some cases, the sample input line of the fluid management system is fluidically coupled to the sample fluid source. The volume range of the sample fluid container can be 1 mL to 100 mL; for example, the volume range of the container can be 1 mL to 90 mL, 1 mL to 80 mL, 1 mL to 70 mL, 1 mL to 60 mL, 1 mL to 50 mL, 1 mL to 40 mL, 1 mL to 30 mL, 1 mL to 20 mL, or 1 mL to 10 mL.

[0095] The sample fluid source may include a sample fluid, and a pressurized sample fluid may flow from the sample fluid source to the flow cell. The sample fluid may be pressurized using a pressurized air source and a sample fluid source air pressure regulator gaseously coupled to the sample fluid source. Air pressure in the sample fluid source may be regulated by a sample fluid pressure feedback control subsystem to control sample fluid pressure and flow rate.

[0096] Fig.14 A flow chart of sub-functions within the main functions of a fluid management system configured to analyze samples according to certain embodiments is provided. The architecture is based on pumps for managing secondary system fluids and all waste. Fluid supply pump 1406 manages primary flush and secondary system fluids 1403. Waste pump 1412 manages waste from drain waste reservoir 1411 and system waste 1414.

[0097] The fluid architecture has positive pressure-based sheath fluid and sample delivery, where the positive pressure source is used to control the ratio of sheath fluid / sample fluid flow rates. The working fluid (sheath fluid) is controlled via regulated air pressure 1402, and the sample fluid pressure is controlled via regulated air pressure 1404. The feedback control 1401 of air pressure is a closed loop implemented via inputs from the working fluid (sheath fluid) chamber 1405 and the sample chamber 1407. Examples of feedback parameters are measured values ​​of chamber air pressure, liquid level height for pressure head compensation, etc. In addition, the working fluid (sheath fluid) pressure control signal is used as feedback for sample pressure control. The sample pressure set point is always the sheath fluid pressure + required flow rate. This is because the sample pressure will always be greater than the working fluid (sheath fluid) pressure of the positive flow through the system. This allows dynamic adjustment of the working fluid without changing the effective pressure drop of the sample fluid.

[0098] The pressurized working fluid (sheath fluid) flows through the fluid supply system 1409 and reaches the flow cell 1408, and the pressurized sample is supplied directly to the flow cell 1408. Fluid in any number of fluid supply reservoirs (e.g., for cleaning fluid) 1403 can also be delivered to the fluid supply system 1409 via the fluid supply pump 1406 in place of the working fluid (sheath fluid) 1405. The dotted lines indicate that (a) the working fluid can be supplied to the flow cell, or alternatively (b) any number of secondary system fluids are supplied to the flow cell. This allows for 100% thorough cleaning of the fluid system. As the working fluid and sample fluid flow through the flow cell and cuvette 1408, the sample is interrogated.

[0099] After the working fluid and sample fluid (or secondary fluid during cleaning) flow through the flow cell, both fluids can be considered "analyzed waste" 1410, which is collected at atmospheric pressure in the exhaust waste reservoir 1411. All waste from the system (analyzed waste 1410 plus any secondary waste sources, such as from cleaning operations 1414) are combined and pumped by a single vacuum waste source (e.g., waste pump 1412). The combined waste is sent to waste storage 1413.

[0100] Fig.15A schematic diagram of a fluid management system according to one embodiment is shown. A pressurized air source 1501 is used to provide air pressure to an electro-pneumatic pressure regulator. A pressure regulator 1502 controls the air pressure in a working fluid reservoir 1504, while a sample fluid regulator 1503 controls the pressure in a sample chamber 1505. A feedback control mechanism 1506 (e.g., a PID controller) is used to send a control signal 1508 to the pressure regulator 1502. A similar feedback control mechanism 1507 is used to send a control signal 1509 to the sample regulator 1503. Feedback for the working fluid pressure comes from the measured air pressure in the working fluid reservoir 1510 and the liquid height in the working fluid reservoir 1513 measured using a level measurement device 1512. Feedback for the sample fluid pressure comes from the measured air pressure in the sample chamber 1511 and the working fluid control signal 1508. The set point of the sample fluid is always controlled to be an increment above the working fluid pressure so that the pressure drop across the sample line is constant even if the working fluid pressure changes. The working fluid reservoir 1504 has a quick disconnect connection 1515 that connects to a conduit 1516 and a mating connector 1514. Any number of secondary fluids are stored in the secondary fluid reservoir set 1553. In addition, the primary flush fluid reservoir 1552 includes the primary flush fluid. As described above, the flow control circuit 1554 selectively allows fluid from the primary flush fluid reservoir 1552 and the secondary fluid reservoir 1553.

[0101] If a primary flush fluid or secondary fluid is required, such as for cleaning an instrument, the connector 1514 and the conduit 1516 can be disconnected from the working fluid supply tank and connected to another mating connector 1518, which is connected to a fluid supply pump 1555. Other fluid supply systems of the present invention described herein may also be used instead. If a sample is being analyzed, the working fluid and the sample fluid meet in the flow cell 1521, and the sample is focused in a hydrodynamic manner into a cuvette 1522, where the sample is interrogated. After analysis, the combined working fluid and sample can be considered as analyzer waste. The waste flows to a discharge waste accumulator 1525 at atmospheric pressure. The accumulator 1525 is at atmospheric pressure to maintain a constant pressure drop on the flow cell and the cuvette. A single waste pump 1530 provides a vacuum to pump all waste sources in the system. The analysis waste in the discharge waste accumulator 1525 is drawn out to a waste management system 1527 through a conduit 1526 having a known fluid resistance. The resistor can be a section of tubing with a known length and inner diameter, an orifice, etc. Any number of additional system waste sources 1528 (e.g., waste sources for cleaning other parts of the instrument) are also connected to the waste management system 1527 via sections of tubing with known fluid resistance 1529. The waste management system can be any fluid management system that combines multiple waste sources in parallel into a single output. The single waste output of the waste management system 1527 is used to pull all waste sources via a waste pump 1530, and all system waste is stored in a waste tank 1531.

[0102] In some embodiments, the fluid management system includes a particle sorter fluid management system, which belongs to a particle sorter and is configured to analyze and sort particles in a sample fluid. After analyzing the particles in the sample fluid, the analyzed sample fluid can flow from the output end of the cuvette to a sorting block coupled to the output end of the cuvette. The flow stream can flow out of a nozzle in the sorting block having a nozzle diameter. In order to generate droplets, the nozzle can be rapidly vibrated by an acoustic device (e.g., a piezoelectric element). In order to sort the particles in the sample, a droplet charging mechanism can charge droplets of the flow stream containing the type of particles to be sorted, for example, at an interruption point in the flow stream. The droplets can pass through an electrostatic field and be deflected into one or more collection containers based on the polarity and magnitude of the charge on the droplets. Uncharged droplets are not deflected by the electrostatic field. Droplets (whether charged or uncharged) can be collected in one or more sample collection containers or collection subsystems, which are appropriately guided (e.g., positioned) to collect one or more flow streams generated by the deflection plate.

[0103] In such embodiments, the fluid management system can be configured to analyze the sample fluid in the cuvette in the same manner as in the particle analyzer, as described in detail above, where, for example, the system analyzes the detected light to measure the physical and fluorescent properties of the particles. The flow cytometer can further sort the particles based on these measured properties. The fluid management system of the particle sorter can be the same as the fluid management system of the particle analyzer, except that the particle sorter fluid management system can include a sorting block, a sorter waste liquid collection subsystem (rather than an analyzer waste liquid collection subsystem) and a sample collection system including one or more sample collection containers. The sorting block can be configured to be coupled (e.g., detachably coupled) to the output end of the cuvette. The sorter waste liquid collection subsystem can collect waste liquid from the sorting, and the sorter waste liquid collection subsystem is in a droplet receiving relationship with the output end of the sorting block. The waste liquid collected by the sorter waste liquid collection subsystem can be transported through the waste liquid management subsystem to the waste liquid container by a fluid moving device. The waste liquid collected by the sorter waste liquid collection subsystem can be sucked into the waste liquid container through the waste liquid management subsystem by the vacuum generated by the fluid moving device. The waste liquid generated by the sorting operation may include, for example, particles that are not sorted into the sample collection container and analyzed fluids (e.g., sample fluids, working fluids).

[0104] As described above, in some cases, the fluid management system includes a sorter waste collection subsystem that is configured to fluidically couple an output end of a sorting block coupled to an output end of a cuvette to a waste management subsystem. The sorter waste collection subsystem may be configured to receive and collect waste liquid from a sorting operation performed by a flow-type particle analyzer. The sorter waste collection subsystem may be configured to receive and collect waste liquid from an output end of the sorting block, for example, waste liquid that has flowed through the cuvette and left a sorting block coupled to the cuvette. The sorter waste collection subsystem may be in a droplet receiving relationship with the sorting block. The sorter waste collection subsystem may include any container and fluid line suitable for collecting waste liquid generated by the sorted sample.

[0105] In some cases, the sorter waste liquid collection subsystem includes a waste liquid aspirator for receiving waste liquid from the output end of the sorting block, and the waste liquid aspirator is fluidically coupled to the waste liquid management subsystem. The waste liquid aspirator may include any suitable components, such as tubes, containers, fluid pipelines, etc., for receiving and conveying waste liquid from the sorting block. The waste liquid aspirator can transfer the waste liquid received from the sorting block to the waste liquid management subsystem. In some cases, the waste liquid in the waste liquid aspirator is sucked into the waste liquid management subsystem by the vacuum generated by the fluid moving device that is fluidically coupled to the waste liquid management subsystem. The waste liquid aspirator can be configured to receive waste liquid from the output end of the sorting block. In some cases, the waste liquid aspirator is in a droplet receiving relationship with the output end of the sorting block coupled to the cuvette. The waste liquid aspirator can be fluidically coupled to the waste liquid management subsystem using a known fluid resistance. In some cases, the fluid pipeline that couples the waste liquid aspirator to the waste liquid management subsystem includes a resistor. The resistor can provide a known fluid resistance in a fluid line coupling the waste aspirator and the waste management subsystem. The resistor can be any resistor described herein.

[0106] In some cases, the fluid management system includes a sorting block that is configured to be removably coupled to the output of the cuvette (e.g., a removable sorting block). "Removably coupled" means that the sorting block can be coupled to (e.g., attached to) the cuvette and then subsequently decoupled from the cuvette at some point, such as disassembled. In some cases, the sorting block is configured to be manually coupled to the output of the cuvette. In such embodiments, the sorting block may have a connector or accessory assembly that allows the sorting block to be coupled to the output of the cuvette. In some cases, the sorting block is configured to be removably coupled to the output of the cuvette in the same manner as the analyzer waste liquid subsystem connector. For example, the sorting block and the analyzer waste liquid collection subsystem connector may both have the same manual connector that is configured to be removably coupled to the output of the cuvette.

[0107] The sorting block may include any suitable components capable of sorting particles in a sample, such as a sorting nozzle and one or more deflection plates. To sort cells by electrostatic methods, the desired cells must be contained in charged droplets. In order to generate droplets, the sorting block (e.g., a sorting nozzle) can be rapidly vibrated by an acoustic device (e.g., a piezoelectric element). The volume of the droplet can be estimated by the fluid dynamics of the flow stream and the nozzle size. In order to charge the droplets, the sorting block may include a charging element whose potential can be rapidly changed. Since the cell stream can leave the sorting nozzle in a substantially downward vertical direction, the droplets can also propagate in this direction after formation. Droplets (whether charged or not) can be collected in a sample collection container or by a waste liquid collection subsystem, which is appropriately guided (e.g., positioned) to collect one or more flow streams generated by the deflection plates. Methods and devices for electrostatic cell sorting are described, for example, in U.S. Pat. Nos. 9,952,076 and 9,404,846, the disclosures of which are incorporated by reference in their entirety.

[0108] The sorting block may include any suitable sorting nozzle configured to generate air flow droplets for sorting. The combined working fluid and sample fluid may flow from the cuvette to the sorting nozzle and out through the orifice of the sorting nozzle in the form of a flow stream. The nozzle orifice may have any diameter, such as 50 μm, 70 μm, 100 μm, or any other suitable diameter. The nozzle diameter may affect the properties of the flow stream, such as stream size, droplet break point, and droplet volume. The flow stream may be a continuous fluid stream or a series of droplets, depending on the action of the droplet generator.

[0109] The sorting block may include any suitable droplet deflector configured to deflect particles in a flow stream flowing therebetween. The flow stream may be a series of droplets that are partially deflected by a pair of deflection plates and turned into multiple streams. Charged droplets may be deflected by a potential applied to the deflection plates and directed away from their original path, such as toward a collection container. Droplets that are neither positively nor negatively charged may not be deflected by the potential applied to the deflection plates and therefore continue to flow along their original path. In some cases, the charged droplets are collected by a waste collection subsystem. In some cases, the uncharged droplets are collected by a waste collection subsystem.

[0110] The sorting block can transport the sorted particles to a sample collection system or a sorter waste collection subsystem. The sorting block can transport the sorted sample (e.g., sorted particles of interest) to a sample collection container. The collection container can be in a droplet receiving relationship with the sorting block; for example, the collection container can receive flow cytometry sorted cell products (e.g., in the form of droplets) from the sorting block. In some cases, the sorting block transports waste liquid to a waste liquid aspirator of the sorter waste liquid collection subsystem. The waste liquid aspirator can be in a droplet receiving relationship with the sorting block; for example, the waste liquid aspirator can receive waste liquid (e.g., in the form of droplets) from the sorting block.

[0111] In some cases, the fluid management system includes a sample collection system including a sample collection container for receiving the sorted sample from the output end of the sorting block. The sample collection container can be any suitable container (e.g., having rigid or flexible walls) for receiving the sorted particles. The sample collection system can be configured to collect the sorted particles in a sterile manner. In some cases, the sample collection system includes, for example, any collection system described in U.S. Publication No. 2019 / 0331657, which is incorporated herein by reference in its entirety.

[0112] Fig.16 A flow chart of the main functions of a fluid management system configured to analyze and sort samples according to certain embodiments is provided. The main functions are: 1601 supplying working fluid (e.g., sheath fluid) and sample fluid to a flow cell in controlled proportions; 1602 supplying any number of primary flushes and / or secondary fluids (e.g., for cleaning); 1603 hydrodynamic focusing of a sample using a working fluid for sample interrogation through a cuvette; collecting unsorted waste; and 1605 managing and storing all system waste (waste from particle sorting 1604 and secondary system waste 1606). The dashed lines indicate that (a) working fluid and sample fluid can be supplied to the flow cell, or alternatively, (b) any number of primary flushes or secondary system fluids can be supplied to the flow cell.

[0113] Fig.17 A flow chart of sub-functions within the main functions of a fluid management system configured to analyze and sort samples according to certain embodiments is provided. The architecture is based on pumps for managing secondary system fluids and all waste. Fluid supply pump 1706 manages primary flush and secondary system fluids 1703. Waste pump 1714 manages all waste from particle sorting 1712 and system waste 1713.

[0114] The fluid architecture has positive pressure-based sheath fluid and sample delivery, where the positive pressure source is used to control the ratio of sheath fluid / sample fluid flow rates. The working fluid (sheath fluid) is controlled via regulated air pressure 1702, and the sample fluid pressure is controlled via regulated air pressure 1704. The feedback control 1701 of air pressure is a closed loop implemented via inputs from the working fluid (sheath fluid) chamber 1705 and the sample chamber 1707. Examples of feedback parameters include measured values ​​of chamber air pressure, liquid level height for pressure head compensation, etc. In addition, the working fluid (sheath fluid) pressure control signal is used as feedback for sample pressure control. The sample pressure set point is always the sheath fluid pressure + required flow rate. This is because the sample pressure will always be greater than the working fluid (sheath fluid) pressure of the positive flow through the system. This allows dynamic adjustment of the working fluid without changing the effective pressure drop of the sample fluid.

[0115] The pressurized working fluid (sheath) flows through the fluid supply system 1709 and reaches the flow cell 1708, and the pressurized sample is directly supplied to the flow cell 1708. The fluid in any number of primary flushing fluids and secondary fluid reservoirs (e.g., for cleaning fluids) 1703 can also be delivered to the fluid supply system 1709 via the fluid supply pump 1706, replacing the working fluid (sheath) 1705. The dotted lines indicate that (a) the working fluid can be supplied to the flow cell, or alternatively, (b) any number of primary flushing or secondary system fluids are supplied to the flow cell. This allows for 100% thorough cleaning of the fluid system. As the primary and sample fluids flow through the flow cell and the cuvette 1708, the sample is interrogated.

[0116] After the working fluid and sample fluid flow through the flow cell and cuvette for analysis, the fluid then flows to the sorting nozzle 1710, which produces a gas stream of droplets. These droplets can be sorted for collection 1711 or waste 1712 via the instrument sorting mechanism. All waste from the system (sorted waste 1712 plus any secondary waste sources, such as from cleaning operations 1713) are combined and pumped by a single vacuum waste source (e.g., waste pump 1714). The combined waste is sent to waste storage 1715.

[0117] Fig.18A schematic diagram of a sorter architecture of a flow cytometer according to one embodiment is shown. A pressurized air source 1801 is used to provide air pressure to an electro-pneumatic pressure regulator. A pressure regulator 1802 controls the air pressure in a working fluid reservoir 1804, while a sample fluid regulator 1803 controls the pressure in a sample chamber 1805. A feedback control mechanism 1806 (e.g., a PID controller) is used to send a control signal 1808 to the pressure regulator 1802. A similar feedback control mechanism 1807 is used to send a control signal 1809 to the sample regulator 1803. Feedback for the working fluid pressure comes from the measured air pressure in the working fluid reservoir 1810 and the liquid height in the working fluid reservoir 1813 measured using a liquid level measurement device 1812. Feedback for the sample fluid pressure comes from the measured air pressure in the sample chamber 1811 and the working fluid control signal 1808. The set point of the sample fluid is always controlled to be an increment above the working fluid pressure so that the pressure drop on the sample line is constant even if the working fluid pressure changes. The working fluid reservoir 1804 has a quick disconnect connection 1815 that connects to a mating connector 1814 .

[0118] Also included is a fluid supply system of the present invention. Any number of secondary fluids are stored in a secondary fluid reservoir 1853. In addition, a primary flushing fluid reservoir 1852 is also included. If a primary flushing fluid or a secondary fluid is needed, such as for cleaning an instrument, the fluid supply connector 1814 and the conduit 1816 can be disconnected from the working fluid supply tank and connected to another mating connector 1818, which is connected to a fluid supply pump 1855. Other fluid supply systems of the present invention described herein may be used instead. The pump can then supply fluid according to the settings of the flow control loop 1854. The fluid is directed to a flow cell 1821. If a sample is being analyzed, the working fluid and the sample fluid meet in the flow cell 1821, and the sample is focused in a fluid dynamics manner into a cuvette 1822, where a sample interrogation is performed. After analysis, the working fluid and the sample pass through a sorting nozzle 1823, which produces air flow droplets for sorting. The sorted samples are collected in the sample collection 1832, and the unsorted waste liquid is collected in the sorting waste liquid aspirator 1833. A single waste liquid pump 1830 provides vacuum to aspirate all waste liquid sources in the system. The sorting waste liquid in the sorting waste liquid aspirator 1833 is drawn out to the waste liquid management system 1827 through a tube with a known fluid resistor 1834. The resistor can be a section of pipe with a known length and inner diameter, an orifice, etc. Any number of additional system waste liquid sources 1828 (for example, waste liquid sources for cleaning other parts of the instrument) are also connected to the waste liquid management system 1827 via a tube section with a known fluid resistance 1829. The waste liquid management system can be any fluid management system that merges multiple waste liquid sources in parallel to a single output end. The single waste liquid output end of the waste liquid management system 1827 is used to pull all waste liquid sources via the waste liquid pump 1830, and all system waste liquids are stored in the waste liquid tank 1831.

[0119] In some embodiments, the fluid management system includes a mixing system. The mixing system can be configured to (a) analyze particles in a sample fluid according to any embodiment described herein (e.g., operate in an analyzer mode), or alternatively, (b) analyze and sort particles in a sample fluid according to any embodiment described herein (e.g., operate in a sorter mode). The mixing system includes: an analyzer waste collection subsystem configured to fluidically couple the output end of the cuvette to a waste management subsystem; and a sorter waste collection subsystem configured to fluidically couple the output end of a sorting block coupled to the output end of the cuvette to the waste management subsystem. The fluid management system can be configured so that in some cases (a) the analyzer waste collection subsystem fluidically couples the output end of the cuvette to the waste management subsystem, or alternatively in some cases (b) the sorter waste collection subsystem fluidically couples the output end of the sorting block coupled to the output end of the cuvette to the waste management subsystem. The fluid management system can be configured to enable a user to select whether the analyzer waste collection subsystem fluidically couples the output end of the cuvette to the waste management subsystem or the sorter waste collection subsystem fluidically couples the output end of the sorting block coupled to the output end of the cuvette to the waste management subsystem. The user can make a selection based on whether he wants the flow-type particle analyzer to (a) analyze the sample or (b) analyze and sort the sample. For example, to enable analyzer mode, a user can manually couple the analyzer waste collection subsystem connector to the output end of the cuvette, where the connector fluidically couples the cuvette to the analyzer waste collection subsystem. In analyzer mode, the sorting block may not be coupled to the output end of the cuvette (or may be detached from it). To enable sorter mode, a user can manually couple the sorting block to the output end of the cuvette and position the sorter waste collection subsystem to be in a droplet receiving relationship with the output end of the sorting block. In some cases, to switch from analyzer mode to sorter mode, the analyzer waste collection subsystem (e.g., analyzer waste collection subsystem connector) is separated from or detached from the cuvette before coupling the sorting block to the cuvette. The hybrid fluid management system may include any elements of the particle analyzer fluid management system and the particle sorter fluid management system as described above.

[0120] Fig.19A flow chart of the main functions of a fluid management system configured to alternately (a) analyze a sample or (b) analyze and sort a sample according to certain embodiments is provided. The main functions are: 1901 supplying a working fluid (e.g., sheath fluid) and a sample fluid to a flow cell in a controlled ratio; 1902 supplying any number of secondary system fluids (e.g., for cleaning); 1903 hydrodynamically focusing a sample using the working fluid for sample interrogation through a cuvette; 1904 collecting analyzed samples as waste if the system is configured as an analyzer; 1905 collecting unsorted waste if the system is configured as a sorter; and 1906 managing and storing all system waste (waste from particle analysis 1904, particle sorting 1905, and system waste 1907). The dashed line before the sample interrogation indicates that (a) the working fluid and the sample fluid can be supplied to the flow cell, or alternatively (b) any number of secondary system fluids can be supplied to the flow cell. The dashed line after sample interrogation indicates that (a) the analyzed sample can be sorted, or alternatively, (b) waste from the analyzed sample is collected without sorting.

[0121] Fig. 20 A flow chart of sub-functions in the main functions of a fluid management system configured to alternately (a) analyze a sample or (b) analyze and sort a sample according to certain embodiments is provided. The architecture is based on pumps for managing secondary system fluids and all wastes. The fluid supply pump 2006 manages the primary flush and secondary system fluids 2003. The waste pump 2016 manages system waste 2015, as well as waste from draining the waste reservoir 2013 and particle sorting 2014.

[0122] The fluid structure has a sheath liquid and sample delivery based on positive pressure, where the positive pressure source is used to control the ratio of the sheath liquid / sample liquid flow rate. The working fluid (sheath liquid) is controlled via the regulated air pressure 2002, and the sample fluid pressure is controlled via the regulated air pressure 2004. The feedback control 2001 of the air pressure is a closed loop implemented via inputs from the working fluid (sheath liquid) chamber 2005 and the sample chamber 2007. Examples of feedback parameters include measured values ​​of chamber air pressure, liquid level height for pressure head compensation, etc. In addition, the working fluid (sheath liquid) pressure control signal is used as feedback for sample pressure control. The sample pressure set point is always the sheath liquid pressure + the required flow rate. This is because the sample pressure will always be greater than the working fluid (sheath liquid) pressure of the positive flow through the system. This allows the working fluid to be dynamically adjusted without changing the effective pressure drop of the sample fluid.

[0123] The pressurized working fluid (sheath fluid) flows through the fluid supply system 2009 and reaches the flow cell 2008, and the pressurized sample is supplied directly to the flow cell 2008. Fluid in any number of fluid supply tanks (e.g., for cleaning fluid) 2003 can also be delivered to the fluid supply system 2009 via the fluid supply pump 2006, replacing the working fluid (sheath fluid) 2005. The dotted lines leading to the fluid supply system indicate that (a) the working fluid can be supplied to the flow cell, or alternatively, (b) any number of secondary system fluids can be supplied to the flow cell. This allows for 100% thorough cleaning of the fluid system. As the working fluid and sample fluid flow through the flow cell and cuvette 2008, the sample is interrogated.

[0124] The mixed fluid architecture includes both the exhaust waste reservoir 2013 from the analysis system, and the sorting nozzle 2011, sample collection 2012 and sorting waste collection 2014 from the sorter architecture. The user can select the operating mode to be performed: only analyze samples that do not require flow generation, or analyze and sort in the sorter. If the system is configured as an analyzer, the waste connection 2010 (such as a connector or joint) can be coupled to a cuvette, where the waste connection fluidically couples the exhaust waste reservoir to the cuvette. If the system is configured as a sorter, the sorting nozzle 2011 can be coupled to the cuvette. The dashed lines after the sample interrogation in the flow cell and cuvette indicate that (a) the analyzed sample can be sorted (2011), or alternatively (b) the waste from the analyzed sample can be collected without sorting (2010). All waste from the system (analyzed waste (2013) or sorted waste (2014) plus any secondary waste sources, such as waste from cleaning operations (2015)) are combined and pumped by a single vacuum waste source (e.g., a waste pump) (2016). The combined waste is sent to waste storage 2017.

[0125] Fig.21A hybrid structure according to one embodiment is shown, including a combination of an analyzer architecture and a sorter architecture, wherein components allow switching between functions. A pressurized air source 2101 is used to provide air pressure to an electro-pneumatic pressure regulator. The pressure regulator 2102 controls the air pressure in the working fluid reservoir 2104, while the sample fluid regulator 2103 controls the pressure in the sample chamber 2105. A feedback control mechanism 2106 (e.g., a PID controller) is used to send a control signal 2108 to the pressure regulator 2102. A similar feedback control mechanism 2107 is used to send a control signal 2109 to the sample regulator 2103. Feedback for the working fluid pressure comes from the measured air pressure in the working fluid reservoir 2110 and the liquid height in the working fluid reservoir 2113 measured using a liquid level measuring device 2112. Feedback for the sample fluid pressure comes from the measured air pressure in the sample chamber 2111 and the working fluid control signal 2108. The set point of the sample fluid is always controlled to be an increment above the working fluid pressure so that the pressure drop across the sample line is constant even if the working fluid pressure changes. The working fluid reservoir 2104 has a quick disconnect connection 2115 that connects to a mating connector 2114 .

[0126] The system also includes a fluid supply system of the present invention. Any number of secondary fluids are stored in the secondary fluid reservoir 2153. A main flushing fluid reservoir 2152 is also included. If a main flush or secondary fluid is needed, such as for cleaning an instrument, the fluid supply connector 2114 and the pipeline 2116 can be disconnected from the working fluid reservoir and connected to another mating connector 2118 connected to the fluid supply pump 2155. Other fluid supply systems of the present invention described herein can be used instead. Then, the pump can supply the main flushing fluid or secondary fluid according to the setting of the flow control loop 2154. The working fluid or secondary fluid is directed to the flow cell 2121. If the sample is being analyzed, the working fluid and the sample fluid meet in the flow cell 2121, and the sample is focused in a fluid dynamics manner into the cuvette 2122, where the sample is interrogated. After analysis, the working fluid and the sample pass through the sorting nozzle 2123, and the sorting nozzle 2123 produces air flow droplets for sorting. The sorted samples are collected in the sample collection 2132, and the unsorted waste liquid is collected in the sorting waste liquid aspirator 2133. If the user wishes to run the system in the analyzer mode only, the sorting nozzle 2123 is removed from the cuvette, and instead, a connector 2124 leading to a discharge waste liquid accumulator 2125 under atmospheric pressure is coupled to the cuvette. The accumulator 2125 is under atmospheric pressure to maintain a constant pressure drop between the circulation pool and the cuvette. A single waste liquid pump 2130 provides vacuum to suck all waste liquid sources in the system. The sorting waste liquid in the sorting waste liquid aspirator 2133 is drawn out to the waste liquid management system 2127 through a pipeline with a known fluid resistor 2134. The analyzer waste liquid discharged in the waste liquid accumulator 2125 is drawn out to the waste liquid management system 2127 through a pipeline with a known fluid resistor 2126. These resistors can be a section of pipe, an orifice, etc. with a known length and inner diameter. Any number of additional system waste sources 2128 (e.g., waste sources for cleaning other parts of the instrument) are also connected to the waste management system 2127 via a tubing segment with a known fluid resistance 2129. The waste management system can be any fluid management system that combines multiple waste sources in parallel into a single output. The single waste output of the waste management system 2127 is used to pull all waste sources through a waste pump 2130, and all system waste is stored in a waste tank 2131.

[0127] Flow cytometry

[0128] Aspects of the present invention also include flow cytometers, which include the fluid supply system of the present invention. The flow cytometer of interest includes, for example, a flow cell configured to transport microparticles in a flow stream, a light source configured to illuminate the flow cell at an interrogation point, and a fluid supply system of the present invention. As described herein, a "flow cell" is described in its conventional sense to refer to a component such as a cuvette, which includes a flow channel having a liquid flow stream for transporting microparticles in a sheath fluid. The cuvette of interest includes a container having a passage therethrough. The flow stream may include a liquid sample injected from a sample tube. The flow cell of interest includes a light-accessible flow channel. In some cases, the flow cell includes a transparent material (e.g., quartz) that allows light to pass through. In some embodiments, the flow cell is an airflow flow cell, wherein the optical interrogation of the microparticles occurs outside the flow cell (i.e., in free space).

[0129] In some cases, the flow stream is configured to be illuminated with light from a light source at an interrogation point. The flow stream for which the flow channel is configured may include a liquid sample injected from a sample tube. In some embodiments, the flow stream may include a narrow, fast-flowing liquid stream arranged so that linearly separated particles transported therein are separated from each other in a single-file manner. The "interrogation point" discussed herein refers to an area within a flow cell where particles are illuminated by light from a light source, such as for analysis. The size of the interrogation point can vary as desired. For example, when 0 μm represents the axis of light emitted by the light source, the interrogation point can range from -100 to 1000, such as -500 to 5000, such as -250 to 4050, and including -15 to 305.

[0130] After the particles are illuminated in the flow cell, particle modulated light can be observed. "Particle modulated light" refers to light received from particles in a flow stream after the particles are illuminated with light from a light source. In some cases, the particle modulated light is side scattered light. As described herein, side scattered light refers to light refracted and reflected from the surface and internal structure of the particles. In additional embodiments, the particle modulated light includes forward scattered light (i.e., light that passes through or bypasses the particles primarily in a forward direction). In other cases, the particle modulated light includes fluorescence (i.e., light emitted from a fluorescent dye after illumination with light of an excitation wavelength).

[0131] As described above, aspects of the present invention also include a light source that is configured to irradiate microparticles passing through a flow cell at an interrogation point. Any convenient light source can be used as the light source described herein. In some embodiments, the light source is a laser. In embodiments, the laser can be any convenient laser, such as a continuous wave laser. For example, the laser can be a diode laser, such as an ultraviolet diode laser, a visible diode laser, and a near-infrared diode laser. In other embodiments, the laser can be a helium-neon (HeNe) laser. In some cases, the laser is a gas laser, such as a helium-neon laser, an argon laser, a krypton laser, a xenon laser, a nitrogen laser, a CO2 laser, a CO laser, an argon-fluorine (ArF) excimer laser, a krypton-fluorine (KrF) excimer laser, a xenon-chlorine (XeCl) excimer laser, or a xenon-fluorine (XeF) excimer laser or a combination thereof. In other cases, the subject flow cytometer includes a dye laser, such as a stilbene, coumarin, or rhodamine laser. In yet other cases, the laser of interest includes a metal vapor laser, such as a helium cadmium (HeCd) laser, a helium mercury (HeHg) laser, a helium selenium (HeSe) laser, a helium silver (HeAg) laser, a strontium laser, a neon copper (NeCu) laser, a copper laser, or a gold laser, and combinations thereof. In yet other cases, the subject flow cytometer includes a solid-state laser, such as a ruby ​​laser, a Nd:YAG laser, a NdCrYAG laser, an Er:YAG laser, a Nd:YLF laser, a Nd:YVO4 laser, a Nd:YCa4O(BO3)3 laser, a Nd:YCOB laser, a titanium sapphire laser, a thulium YAG laser, a ytterbium YAG laser, a ytterbium2O3 laser, or a cerium-doped laser, and combinations thereof.

[0132] The laser light source according to some embodiments may also include one or more optical adjustment components. In some embodiments, the optical adjustment component is located between the light source and the flow cell, and may include any device capable of changing the spatial width of the irradiation or some other characteristics of the irradiation from the light source, such as the irradiation direction, wavelength, beam width, beam intensity, and focus. The optical adjustment protocol may include any convenient device for adjusting one or more characteristics of the light source, including but not limited to lenses, mirrors, filters, optical fibers, wavelength separators, pinholes, slits, collimation protocols, and combinations thereof. In some embodiments, the flow cytometer of interest includes one or more focusing lenses. In one example, the focusing lens can be a reduction lens. In other embodiments, the flow cytometer of interest includes an optical fiber.

[0133] When the optical adjustment component is configured to move, the optical adjustment component can be configured to move continuously or in discrete intervals, for example in increments of 0.01 mm or greater, for example 0.05 mm or greater, for example 0.1 mm or greater, for example 0.5 mm or greater, for example 1 mm or greater, for example 10 mm or greater, for example 100 mm or greater, for example 500 mm or greater, for example 1 mm or greater, for example 5 mm or greater, for example 10 mm or greater, and including increments of 25 mm or greater.

[0134] Any displacement protocol can be used to move the optical adjustment assembly structure, such as coupled to a movable support table or directly coupled to a motor-actuated translation stage, a screw translation assembly, a gear translation device, such as those using stepper motors, servo motors, brushless motors, brushed DC motors, micro-step drive motors, high-resolution stepper motors, and other types of motors.

[0135] The light source can be located at any suitable distance from the flow cell, for example, the light source is separated from the flow cell by 0.005 mm or more, for example, 0.01 mm or more, for example, 0.05 mm or more, for example, 0.1 mm or more, for example, 0.5 mm or more, for example, 1 mm or more, for example, 5 mm or more, for example, 10 mm or more, for example, 25 mm or more, including a distance of 100 mm or more. In addition, the light source can be located at any suitable angle relative to the flow cell, for example, an angle in the range of 10 to 90 degrees, for example, 15 to 85 degrees, for example, 20 to 80 degrees, for example, 25 to 75 degrees, including 30 to 60 degrees, for example, at an angle of 90 degrees.

[0136] In some embodiments, the light source of interest includes a plurality of lasers configured to provide laser light to discretely illuminate the flow stream, such as 2 lasers or more, such as 3 lasers or more, such as 4 lasers or more, such as 5 lasers or more, such as 10 lasers or more, and including 15 lasers or more, which are configured to provide laser light to discretely illuminate the flow stream. Depending on the desired wavelength of light for illuminating the flow stream, each laser can have a specific wavelength that varies from 200nm to 1500nm, such as from 250nm to 1250nm, such as from 300nm to 1000nm, such as from 350nm to 900nm and including from 400nm to 800nm. In some embodiments, the laser of interest can include one or more of a 405nm laser, a 488nm laser, a 561nm laser, and a 635nm laser.

[0137] As described above, the particle analyzer of interest may also include one or more particle modulated light detectors for detecting particle modulated light intensity data. In some embodiments, the particle modulated light detector includes one or more forward scattered light detectors configured to detect forward scattered light. For example, the target particle analyzer may include 1 forward scattered light detector or more forward scattered light detectors, such as 2 or more, such as 3 or more, such as 4 or more, and including 5 or more. In certain embodiments, the particle analyzer includes 1 forward scattered light detector. In other embodiments, the particle analyzer includes 2 forward scattered light detectors.

[0138] Any convenient detector for detecting collected light may be used for the forward scattered light detector described herein. Detectors of interest may include, but are not limited to, optical sensors or detectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCD), enhanced charge coupled devices (ICCD), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMTs), phototransistors, quantum dot photoconductors or photodiodes and combinations thereof, and other detectors. In certain embodiments, a charge coupled device (CCD), a semiconductor charge coupled device (CCD), an active pixel sensor (APS), a complementary metal oxide semiconductor (CMOS) image sensor, or an N-type metal oxide semiconductor (NMOS) image sensor is used to measure the collected light. In certain embodiments, the detector is a photomultiplier tube, such as a photomultiplier tube, with an effective detection surface area of ​​0.01 cm per region. 2 Up to 10cm 2 Photomultiplier tube, for example, from 0.05cm 2 Up to 9cm 2 , for example, from 0.1cm 2 Up to 8cm 2 , for example from 0.5cm 2 Up to 7cm 2 , and includes from 1cm 2 Up to 5cm 2 .

[0139] In an embodiment, the forward scattered light detector is configured to measure light continuously or at discrete intervals. In some cases, the detector of interest is configured to measure the collected light continuously. In other cases, the detector of interest is configured to measure at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds, and including measuring light every 1000 milliseconds or some other interval.

[0140] In additional embodiments, the one or more particle modulated light detectors may include one or more side scattered light detectors for detecting side scattered light wavelengths (i.e., light refracted and reflected from the surface and internal structure of the particle). In some embodiments, the particle analyzer includes a single side scattered light detector. In other embodiments, the particle analyzer includes multiple side scattered light detectors, such as 2 or more, such as 3 or more, such as 4 or more, and including 5 or more.

[0141] Any convenient detector for detecting collected light can be used in the side scattered light detector described herein. Detectors of interest may include, but are not limited to, optical sensors or detectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCD), enhanced charge coupled devices (ICCD), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMTs), phototransistors, quantum dot photoconductors or photodiodes and combinations thereof, and other detectors. In certain embodiments, a charge coupled device (CCD), a semiconductor charge coupled device (CCD), an active pixel sensor (APS), a complementary metal oxide semiconductor (CMOS) image sensor, or an N-type metal oxide semiconductor (NMOS) image sensor is used to measure the collected light. In certain embodiments, the detector is a photomultiplier tube, such as ... 2 Up to 10cm 2 Photomultiplier tube, for example, from 0.05cm 2 Up to 9cm 2 , for example, from 0.1cm 2 Up to 8cm 2 , for example from 0.5cm 2 Up to 7cm 2 And including from 1cm 2 Up to 5cm 2 .

[0142] In an embodiment, the subject particle analyzer further comprises a fluorescence detector configured to detect light of one or more fluorescent wavelengths. In other embodiments, the particle analyzer comprises a plurality of fluorescence detectors, such as 2 or more, such as 3 or more, such as 4 or more, such as 5 or more, such as 10 or more, such as 15 or more, and including 20 or more.

[0143] Any convenient detector for detecting collected light can be used in the fluorescence detector described herein. Detectors of interest may include, but are not limited to, optical sensors or detectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCD), enhanced charge coupled devices (ICCD), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMTs), phototransistors, quantum dot photoconductors or photodiodes and combinations thereof, and other detectors. In certain embodiments, a charge coupled device (CCD), a semiconductor charge coupled device (CCD), an active pixel sensor (APS), a complementary metal oxide semiconductor (CMOS) image sensor, or an N-type metal oxide semiconductor (NMOS) image sensor is used to measure the collected light. In certain embodiments, the detector is a photomultiplier tube, such as a photomultiplier tube, with an effective detection surface area of ​​0.01 cm per region. 2 Up to 10cm 2 Photomultiplier tube, for example, from 0.05cm 2 Up to 9cm 2 , for example, from 0.1cm 2 Up to 8cm 2 , for example from 0.5cm 2 Up to 7cm 2 , and includes from 1cm 2 Up to 5cm 2 .

[0144] When the subject particle analyzer includes multiple fluorescence detectors, each fluorescence detector can be identical, or the collection of fluorescence detectors can be a combination of different types of detectors. For example, when the subject particle analyzer includes two fluorescence detectors, in some embodiments, the first fluorescence detector is a CCD type device, and the second fluorescence detector (or imaging sensor) is a CMOS type device. In other embodiments, both the first and second fluorescence detectors are CCD type devices. In other embodiments, both the first and second fluorescence detectors are CMOS type devices. In other embodiments, the first fluorescence detector is a CCD type device, and the second fluorescence detector is a photomultiplier tube (PMT). In other embodiments, the first fluorescence detector is a CMOS type device, and the second fluorescence detector is a photomultiplier tube. In other embodiments, both the first and second fluorescence detectors are photomultiplier tubes.

[0145] In embodiments of the present disclosure, the fluorescence detector of interest is configured to measure collected light at one or more wavelengths, such as at 2 or more wavelengths, such as at 5 or more different wavelengths, such as at 10 or more different wavelengths, such as at 25 or more different wavelengths, such as at 50 or more different wavelengths, such as at 100 or more different wavelengths, such as at 200 or more different wavelengths, such as at 300 or more different wavelengths, and including measuring light emitted by a sample in flow at 400 or more different wavelengths. In some embodiments, 2 or more detectors in the particle analyzer described herein are configured to measure collected light of the same or overlapping wavelengths.

[0146] In some embodiments, the fluorescence detector of interest is configured to measure the collected light within a certain wavelength range (e.g., 200nm is configured to measure). In some embodiments, the detector of interest is configured to collect the spectrum of light within a certain wavelength range. For example, the particle analyzer may include one or more detectors configured to collect the spectrum of light within one or more wavelength ranges. In some other embodiments, the detector of interest is configured to measure the light emitted by the sample in the flow stream at one or more specific wavelengths. For example, the particle analyzer may include one or more detectors configured to measure light at one or more of the following wavelengths: 450nm, 518nm, 519nm, 561nm, 578nm, 605nm, 607nm, 625nm, 650nm, 660nm, 667nm, 670nm, 668nm, 695nm, 710nm, 723nm, 780nm, 785nm, 647nm, 617nm and any combination thereof. In certain embodiments, one or more detectors may be configured to pair with a specific fluorophore, such as a fluorophore to be used with a sample in a fluorescence assay.

[0147] In some embodiments, the particle analyzer includes one or more wavelength separators, which are located between the flow cell and the particle modulated light detector. The term "wavelength separator" is used in its conventional sense herein and refers to an optical component configured to separate the light collected from the sample into a predetermined spectral range. In some embodiments, the particle analyzer includes a single wavelength separator. In other embodiments, the particle analyzer includes a plurality of wavelength separators, such as 2 or more wavelength separators, such as 3 or more, such as 4 or more, such as 5 or more, such as 6 or more, such as 7 or more, such as 8 or more, such as 9 or more, such as 10 or more, such as 15 or more, such as 25 or more, such as 50 or more, such as 75 or more, and includes 100 or more wavelength separators. In some embodiments, the wavelength separator is configured to separate the light collected from the sample into a predetermined spectral range by passing light with a predetermined spectral range and reflecting light of one or more remaining spectral ranges. In other embodiments, the wavelength separator is configured to separate the light collected from the sample into a predetermined spectral range by passing light having a predetermined spectral range and absorbing light of one or more remaining spectral ranges. In other embodiments, the wavelength separator is configured to spatially diffract the light collected from the sample into a predetermined spectral range. Each wavelength separator can be any convenient light separation protocol, such as one or more dichroic mirrors, bandpass filters, diffraction gratings, beam splitters, or prisms. In some embodiments, the wavelength separator is a prism. In other embodiments, the wavelength separator is a diffraction grating. In certain embodiments, the wavelength separator in the subject light detection system is a dichroic mirror.

[0148] Suitable flow cytometry systems may include, but are not limited to, those described in Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford University Press (1997); Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology 91, Humana Press (1997); Practical Flow Cytometry, 3rd ed., Wiley-Liss (1995); Virgo et al. (2012) Ann Clin Biochem. Jan;49(Part 1):17-28; Linden et al., Semin Throm Hemost. 2004 Oct;30(5):502-11; Alison et al., J Pathol, 2010 Dec;222(4):335-344; and Herbig et al. (2007) Crit Rev. Ther Drug Carrier Syst. 24(3): 203-255; the disclosure of which is incorporated herein by reference. In some cases, flow cytometry systems of interest include the BD Biosciences FACSCanto TM Flow cytometer, BD Biosciences FACSCanto TM II flow cytometer, BD Accuri TM Flow cytometer, BD Accuri TM C6 Plus flow cytometer, BD Biosciences FACSCelesta TM Flow cytometer, BD Biosciences FACSLyric TM Flow cytometer, BD Biosciences FACSVerse TM Flow cytometer, BD Biosciences FACSymphony TM Flow cytometer, BD Biosciences LSRFortessa TM Flow cytometer, BD Biosciences LSRFortessa TM X-20 flow cytometer, BD Biosciences FACSPresto TM Flow cytometer, BD Biosciences FACSVia TMFlow cytometer and BD Biosciences FACSCalibur TM Cell sorter, BD Biosciences FACSCount TM Cell sorter, BD Biosciences FACSLyric TM Cell sorter, BD BiosciencesVia TM Cell sorter, BD Biosciences Influxe Cell Sorter, BD Biosciences Jazzc Cell Sorter, BD Biosciences Ariac Cell Sorter, BD Biosciences FACSAriac II Cell Sorter, BD Biosciences FACSAriac III Cell Sorter, BD Biosciences FACSAriac Fusion Cell Sorter, BD Biosciences FACSMelodys Cell Sorter, or BD Biosciences FACSymphony TM S6 cell sorter, etc.

[0149] In some embodiments, the subject system is a flow cytometry system, such as described in U.S. Pat. Nos. 10,663,476, 10,620,111, 10,613,017, 10,605,713, 10,585,031, 10,578,542, 10,578,469, 10,481,074, 10,302,545, 10,145,793, 10,113,967, 10,006,852, 9,952,076, 9,933,341, 9,726,527, 9,453,789, 9,200,334, 9,097,640, 9,989,990,991,983,994,995,996,997,998,9 ... and 4,987,086; and 4,498,766, the disclosures of which are incorporated herein by reference in their entireties.

[0150] In some cases, the flow cytometry systems of the present invention are configured to image microparticles in a flow stream by fluorescence imaging using radio frequency labeled emission (FIRE), such as those described by Diebold et al. in Nature Photonics, Vol. 7(10); 806-810 (2013) and U.S. Pat. Nos. 9,423,353, 9,784,661, 9,983,132, 10,006,852, 10,078,045, 10,036,699, 10,222,316, 10,288,546, 10,324,019, 10, 408,758, 10,451,538, 10,620,111; and those described in U.S. Patent Publication Nos. 2017 / 0133857, 2017 / 0328826, 2017 / 0350803, 2018 / 0275042, 2019 / 0376895, and 2019 / 0376894, the disclosures of which are incorporated herein by reference.

[0151] Fig. 22 A system 2200 for flow cytometry according to an exemplary embodiment of the present invention is shown. The system 2200 includes a flow cytometer 2210, a controller / processor 2290, and a memory 2295. The flow cytometer 2210 includes one or more excitation lasers 2215a-2215c, a focusing lens 2220, a flow chamber 2225, a forward scatter detector 2230, a side scatter detector 2235, a fluorescence collection lens 2240, one or more beam splitters 2245a-2245g, one or more bandpass filters 2250a-2250e, one or more longpass ("pass-through") filters 2255a-2255b, and one or more fluorescence detectors 2260a-2260f.

[0152] The excitation lasers 2215a-2215c emit light in the form of laser beams. Fig. 22 In the example system of , the wavelengths of the laser beams emitted from the excitation lasers 2215a-2215c are 488nm, 633nm and 325nm, respectively. The laser beams are first directed through one or more of the beam splitters 2245a and 2245b. The beam splitter 2245a transmits the 488nm light and reflects the 633nm light. The beam splitter 2245b transmits ultraviolet light (light with a wavelength range of 10 to 400nm) and reflects the 488nm and 633nm light.

[0153] The laser beam is then directed to a focusing lens 2220, which focuses the beam onto the portion of the flow stream where the sample particles are located within a flow cell 2225. The flow cell is the portion of the fluidic system that directs the particles in the stream (usually one at a time) toward the focused laser beam for interrogation. The flow cell may include a flow cell in a benchtop cytometer or a nozzle head in an air flow cytometer.

[0154] Light from the laser beam interacts with particles in the sample through diffraction, refraction, reflection, scattering, and absorption, and is re-emitted at various wavelengths depending on the characteristics of the particles (e.g., their size, internal structure, and the presence of one or more fluorescent molecules attached to or naturally present on or within the particles). Fluorescent emission and diffracted, refracted, reflected, and scattered light can be routed to one or more of a forward scatter detector 2230, a side scatter detector 2235, and one or more fluorescence detectors 2260a-2260f via one or more of beam splitters 2245c-2245g, bandpass filters 2250a-2250e, longpass filters 2255a-2255b, and a fluorescence collection lens 2240.

[0155] Fluorescence collection lens 2240 collects light emitted from the particle-laser beam interaction and directs the light to one or more beam splitters and filters. Bandpass filters (e.g., bandpass filters 2250a-2250e) allow a narrow range of wavelengths to pass through the filter. For example, bandpass filter 2250a is a 510 / 20 filter. The first number represents the center of the spectral band. The second number provides the range of the spectral band. Therefore, a 510 / 20 filter extends 10nm on each side of the center of the spectral band, or extends from 500nm to 520nm. Short-pass filters transmit light with a wavelength equal to or shorter than a specified wavelength. Long-pass filters (e.g., long-pass filters 2255a-2255b) transmit light with a wavelength equal to or longer than a specified wavelength. For example, long-pass filter 2255b is a 670nm long-pass filter that transmits light equal to or longer than 670nm. Filters are typically selected to optimize the specificity of the detector for a particular fluorescent dye. The filters can be configured so that the spectral band of light transmitted to the detector is close to the emission peak of the fluorescent dye.

[0156] The position of the forward scattering detector 2230 is slightly offset from the axis of the direct beam passing through the flow cell, and is configured to detect diffracted light, that is, the excitation light that mainly passes through or bypasses the microparticles in the forward direction. The intensity of the light detected by the forward scattering detector depends on the overall size of the microparticle. The forward scattering detector may include a photodiode. The side scattering detector 2235 is configured to detect refracted and reflected light from the surface and internal structure of the microparticle, which increases as the complexity of the microparticle structure increases. Fluorescence emission from fluorescent molecules associated with the microparticles can be detected by one or more fluorescence detectors 2260a-2260f. The side scattering detector 2235 and the fluorescence detector may include a photomultiplier tube. The signals detected at the forward scattering detector 2230, the side scattering detector 2235 and the fluorescence detector may be converted into electronic signals (voltages) by the detectors. The data may provide information about the sample.

[0157] Those skilled in the art will recognize that the flow cytometer according to embodiments of the present invention is not limited to Fig. 22 The flow cytometer shown in the flow cytometer may include any flow cytometer known in the art. For example, the flow cytometer may have any number of lasers, beam splitters, filters, and detectors, with various wavelengths and various different configurations.

[0158] In operation, the cytometer operation is controlled by the controller / processor 2290, and the measurement data from the detector may be stored in the memory 2295 and processed by the controller / processor 2290. Although not explicitly shown, the controller / processor 2290 is coupled to the detector to receive the output signal from the detector, and may also be coupled to the electrical and electromechanical components of the flow cytometer 2210 to control the laser, flow stream parameters, etc. An input / output (I / O) function 2297 may also be provided in the system. The memory 2295, the controller / processor 2290, and the I / O 2297 may be provided as a whole as an integral part of the flow cytometer 2210. In such an embodiment, the display may also form part of the I / O function 2297 for presenting experimental data to a user of the cytometer 2210. Alternatively, some or all of the memory 2295, the controller / processor 2290, and the I / O function may be part of one or more external devices (e.g., a general purpose computer). In some embodiments, some or all of the memory 2295 and controller / processor 2290 may be in wireless or wired communication with the cytometer 2210. The controller / processor 2290 in conjunction with the memory 2295 and I / O 2297 may be configured to perform various functions related to the preparation and analysis of flow cytometer experiments.

[0159] Fig. 22The system shown in 2297 includes six different detectors that detect fluorescence in six different wavelength bands (which may be referred to herein as the "filter windows" of a given detector), which are defined by the configuration of filters and / or spectrometers in the beam path from the flow cell 2225 to each detector. Different fluorescent molecules used in flow cytometer experiments will emit light in their own characteristic wavelength bands. The specific fluorescent labels used in the experiment and their associated fluorescence emission bands can be selected to be roughly consistent with the filter windows of the detectors. I / O 2297 can be configured to receive data about a flow cytometer experiment, which has a set of fluorescent labels and multiple cell populations with multiple labels, each cell population having a subset of multiple labels. I / O 2297 can also be configured to receive biological data that assigns one or more labels to one or more cell populations, label density data, emission spectrum data, data that assigns labels to one or more labels, and cytometer configuration data. Flow cytometer experiment data (such as label spectral characteristics and flow cytometer configuration data) can also be stored in memory 2295. The controller / processor 2290 may be configured to evaluate one or more assignments of tags to labels.

[0160] In some embodiments, the subject system is a particle sorting system configured to sort particles using a closed particle sorting module, such as those described in U.S. Patent Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosure of which is incorporated herein by reference. In certain embodiments, particles (e.g., cells) of a sample are sorted using a sorting decision module having multiple sorting decision units, such as those described in U.S. Patent Publication No. 2020 / 0256781, filed on December 23, 2019, the disclosure of which is incorporated herein by reference. In some embodiments, a system for sorting components of a sample includes a particle sorting module with a deflection plate, such as those described in U.S. Patent Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosure of which is incorporated herein by reference.

[0161] Fig.23 A functional block diagram of one example of a sorting control system (eg, processor 2300) for analyzing and displaying biological events is shown. Processor 2300 may be configured to implement various processes for controlling the graphical display of biological events.

[0162] The flow cytometer or sorting system 2302 can be configured to acquire biological event data. For example, the flow cytometer can generate flow cytometer event data (e.g., particle modulated light data). The flow cytometer 2302 can be configured to provide the biological event data to the processor 2300. A data communication channel can be included between the flow cytometer 2302 and the processor 2300. The biological event data can be provided to the processor 2300 via the data communication channel.

[0163] Processor 2300 may be configured to receive biological event data from flow cytometer 2302. The biological event data received from flow cytometer 2302 may include flow cytometry event data. Processor 2300 may be configured to provide a graphical display of a first graph including biological event data to display device 2306. Processor 2300 may be further configured to render a region of interest as a gate around a population of biological event data displayed by display device 2306, for example, overlaid on the first graph. In some embodiments, a gate may be a logical combination of one or more graphical regions of interest plotted on a single parameter histogram or a bivariate graph. In some embodiments, the display may be used to display particle parameters or saturation detector data.

[0164] Processor 2300 may also be configured to display the bio-event data within the gate in a manner different from other events in the bio-event data outside the gate on display device 2306. For example, processor 2300 may be configured to render the color of the bio-event data contained within the gate different from the color of the bio-event data outside the gate. Display device 2306 may be implemented as a display, tablet computer, smart phone, or other electronic device configured to present a graphical interface.

[0165] The processor 2300 may be configured to receive a door selection signal for identifying a door from a first input device. For example, the first input device may be implemented as a mouse 2310. The mouse 2310 may initiate a door selection signal to the processor 2300 for identifying a door to be displayed on the display device 2306 or manipulated via the display device 2306 (e.g., by clicking on the desired door when the cursor is on the desired door). In some embodiments, the first device may be implemented as a keyboard 2308 or other device for providing input signals to the processor 2300, such as a touch screen, a stylus, an optical detector, or a voice recognition system. Some input devices may include multiple input functions. In such embodiments, the input functions may each be considered an input device. For example, Fig.23 As shown, the mouse 2310 may include a right mouse button and a left mouse button, each of which may generate a trigger event.

[0166] The triggering event may cause the processor 2300 to change the way the data is displayed, which portions of the data are actually displayed on the display device 2306, and / or provide input for further processing, such as selecting a particle population of interest for particle sorting. In some embodiments, the processor 2300 may be configured to detect when a gate selection is initiated by the mouse 2310. The processor 2300 may be further configured to automatically modify the plot visualization to facilitate the gating process.

[0167] Processor 2300 may be connected to storage device 2304. Storage device 2304 may be configured to receive and store biological event data from processor 2300. Storage device 2304 may also be configured to receive and store flow cytometry event data from processor 2300. Storage device 2304 may further be configured to allow processor 2300 to retrieve biological event data, such as flow cytometry event data.

[0168] The display device 2306 can be configured to receive display data from the processor 2300. The display data can include graphs of the biological event data and gates outlining portions of the graph. The display device 2306 can be further configured to change the information presented based on input received from the processor 2300 and input from the flow cytometer 2302, the storage device 2304, the keyboard 2308, and / or the mouse 2310.

[0169] In some embodiments, the processor 2300 can generate a user interface to receive sample events for sorting. For example, the user interface can include a mechanism for receiving sample events or sample images. The sample events or images or sample gates can be provided before collecting event data for the sample, or can be provided based on an initial set of events for a portion of the sample.

[0170] Fig.24A 2400 (e.g., flow cytometer 2302) according to one embodiment described herein. In some embodiments, particle sorting system 2400 is a cell sorting system. Fig.24A As shown, a droplet formation transducer 2402 (e.g., a piezoelectric oscillator) is coupled to a fluid conduit 2401, which may be coupled to, may include, or may be a nozzle 2403. Within the fluid conduit 2401, a sheath fluid 2404 hydrodynamically focuses a sample fluid 2406 containing particles 2409 into a moving fluid column 2408 (e.g., a stream). In the moving fluid column 2408, the particles 2409 (e.g., cells) are arranged in a line through a monitored area 2411 (e.g., where the laser streams intersect) and are illuminated by a radiation source 2412 (e.g., a laser). The vibration of the droplet formation transducer 2402 causes the moving fluid column 2408 to break into a plurality of droplets 2410, some of which contain the particles 2409.

[0171] In operation, a detection station 2414 (e.g., an event detector) identifies when a target particle (or target cell) passes through the monitored area 2411. The detection station 2414 feeds into a timing circuit 2428, which in turn feeds into a flash charge circuit 2430. At the droplet breakoff point, informed by a timed droplet delay (Δ at), a flash charge can be applied to the moving fluid column 2408 such that the target droplet is charged. The target droplet may include one or more particles or cells to be sorted. The charged droplets may then be sorted by activating a deflection plate (not shown) to deflect the droplet into a container (e.g., a collection tube or a multi-porous or micro-well sample plate), where a hole or micro-well may be associated with a particular target droplet. As Fig.24A As shown, the droplets can be collected in a discharge container 2438.

[0172] A detection system 2416 (e.g., a droplet boundary detector) is used to automatically determine the phase of the droplet drive signal as the target particle passes through the monitored area 2411. An exemplary droplet boundary detector is described in U.S. Patent No. 7,679,039, which is incorporated herein by reference in its entirety. The detection system 2416 allows the instrument to accurately calculate the position of each detected particle in the droplet. The detection system 2416 can feed an amplitude signal 2420 and / or a phase 2418 signal, which in turn feeds (via amplifier 2422) an amplitude control circuit 2426 and / or a frequency control circuit 2424. The amplitude control circuit 2426 and / or the frequency control circuit 2424 in turn control the droplet formation transducer 2402. The amplitude control circuit 2426 and / or the frequency control circuit 2424 can be included in the control system.

[0173] In some embodiments, the sorting electronics (e.g., detection system 2416, detection station 2414, and processor 2440) can be coupled to a memory configured to store detected events and sorting decisions based on the events. The sorting decision can be included in the event data of the particles. In some embodiments, the detection system 2416 and the detection station 2414 can be implemented as a single detection unit or communicatively coupled so that event measurements can be collected by one of the detection system 2416 or the detection station 2414 and provided to a non-collecting element.

[0174] Fig. 24B is a schematic diagram of a particle sorting system according to an embodiment proposed herein. Fig. 24B The particle sorting system 2400 shown in FIG. 2400 includes deflection plates 2452 and 2454. A charge can be applied via a stream of charge wires in the barbs. This produces a droplet stream 2410 containing particles 2409 for analysis. The particles can be illuminated with one or more light sources (e.g., lasers) to produce light scattering and fluorescence information. The particles can be detected by, for example, sorting electronics or other detection systems ( Fig. 24B The deflection plates 2452 and 2454 can be independently controlled to attract or repel charged droplets, thereby directing the droplets to a target collection container (e.g., one of 2472, 2474, 2476, or 2478). Fig. 24B As shown, deflector plates 2452 and 2454 can be controlled to direct the particles along first path 2462 to container 2474, or along second path 2468 to container 2478. If the particle is not a target (e.g., does not show scattering or illumination information within a specified sorting range), the deflector plates can allow the particle to continue to flow along flow path 2464. Such uncharged droplets can enter a waste container, such as via aspirator 2470.

[0175] Sorting electronics may be included to enable collection of measurements, receive fluorescent signals from particles, and determine how to adjust the deflection plates to sort the particles. Fig. 24B An example implementation of the embodiment shown in FIG. 1 includes a BD FACSAria CT scan commercially available from Becton, Dickinson and Company (Franklin Lakes, NJ). TM Serial flow cytometer.

[0176] In some embodiments where the flow cytometer is a cell sorter, the flow cytometer is a particle sorter with imaging capabilities. In such embodiments, the fluid supply system of the present invention can be used to supply working fluid, primary flushing fluid and / or secondary fluid to the flow cell of the particle sorter with imaging capabilities. In some embodiments, the system is a fluorescence imaging particle sorter capable of using radio frequency markers to emit images, such as Fig.25As shown. The particle sorter 2500 includes a light irradiation assembly 2500a, which includes a light source 2501 (e.g., a 488nm laser) that generates an output beam 2501a, which is split into beams 2502a and 2502b using a beam splitter 2502. The beam 2502a propagates through an acousto-optic device (e.g., an acousto-optic deflector, AOD) 2503 to generate an output beam 2503a having one or more angularly deflected beams. In some cases, the output beam 2503a generated from the acousto-optic device 2503 includes a local oscillator beam and a plurality of RF comb beams. The beam 2502b propagates through an acousto-optic device (e.g., an acousto-optic deflector, AOD) 2504 to generate an output beam 2504a having one or more angularly deflected beams. In some cases, the output beam 2504a generated from the acousto-optic device 2504 includes a local oscillator beam and a plurality of RF comb beams. Output beams 2503a and 2504a generated from the acousto-optic devices 2503 and 2504, respectively, are combined with a beam splitter 2505 to generate an output beam 2505a, which is transmitted through an optical component 2506 (e.g., an objective lens) to illuminate the particles in the flow cell 2507. In some embodiments, the acousto-optic device 2503 (AOD) splits a single laser beam into an array of beams, each with a different optical frequency and angle. The second AOD 2504 adjusts the optical frequency of the reference beam, which then overlaps with the beam array at the beam combiner 2505. In some embodiments, the light irradiation system having a light source and an acousto-optic device can also include those systems described by Schraivogel et al. in "High-speed fluorescence image-enabled cell sorting" (Science (2022), 375 (6578): 315-320) and in U.S. Patent Publication No. 2021 / 0404943, the disclosures of which are incorporated herein by reference.

[0177] Output beam 2505a illuminates sample particles 2508 propagating through flow cell 2507 at illumination region 2510 (e.g., using sheath fluid 2509). Sheath fluid 2509 may be supplied to flow cell 2507 by a fluid supply system of the present invention, such as described above. As shown in illumination region 2510, multiple beams (e.g., angle-deflected RF-shifted beams represented by dots on illumination region 2510) overlap with a reference local oscillator beam (represented by hatching on illumination region 2510). Due to their different optical frequencies, the overlapping beams exhibit beat frequency behavior, which causes each beam to emit light at a different frequency f. 1-n Carrying sinusoidal modulation.

[0178] Light from the irradiated sample is transmitted to a light detection system 2500b including a plurality of photodetectors. The light detection system 2500b includes a forward scattered light photodetector 2511 for generating a forward scattered image 2511a and a side scattered light photodetector 2512 for generating a side scattered image 2512a. The light detection system 2500b also includes a bright field photodetector 2513 for generating a light loss image 2513a. In some embodiments, the forward scattering detector 2511 and the side scattering detector 2512 are photodiodes (e.g., avalanche photodiodes, APDs). In some cases, the bright field photodetector 2513 is a photomultiplier tube (PMT). Fluorescence photodetectors 2514-2517 are also used to detect fluorescence from the irradiated sample. In some cases, the photodetectors 2514-2517 are photomultiplier tubes. The light from the irradiated sample is directed to the side scattering detection channel 2512 and the fluorescence detection channel 2514-2517 through a beam splitter 2520. Light detection system 2500b includes bandpass optical components 2521, 2522, 2523, and 2524 (e.g., dichroic mirrors) for transmitting light of predetermined wavelengths to photodetectors 2514-2517. In some cases, optical component 2521 is a 534nm / 40nm bandpass. In some cases, optical component 2522 is a 586nm / 42nm bandpass. In some cases, optical component 2523 is a 700nm / 54nm bandpass. In some cases, optical component 2524 is a 783nm / 56nm bandpass. The first number indicates the center of the spectral band. The second number provides the range of the spectral band. Thus, a 510 / 20 filter extends 10nm on each side of the center of the spectral band, or from 500nm to 520nm.

[0179] Data signals generated in response to light detected in scattered light detection channels 2511 and 2512, bright field light detection channel 2513, and fluorescence detection channels 2514-2517 are digitally processed in real time by processors 2550 and 2551. Based on the data signals generated in processors 2550 and 2551, images 2511a-2517a can be generated in each light detection channel. Image-supported sorting is performed in response to a sorting signal generated in a sorting trigger 2552. The sorting assembly 2500c includes a deflection plate 2531 for deflecting particles into a sample container 2532 or a waste stream 2533. In some cases, the sorting assembly 2500c is configured to sort particles using a closed particle sorting module, such as those described in U.S. Patent Publication No. 2017 / 0299493 filed on March 28, 2017, the disclosure of which is incorporated herein by reference. In some embodiments, the sorting component 2500c includes a sorting decision module having multiple sorting decision units, such as those described in U.S. Patent Publication No. 2020 / 0256781, the disclosure of which is incorporated herein by reference.

[0180] Fig.26 The general architecture of an example computing device 2600 is shown in accordance with some embodiments. Fig.26 The general architecture of the computing device 2600 shown in includes the arrangement of computer hardware and software components. However, in order to provide an achievable disclosure, it is not necessary to display all of these generally conventional elements. As shown, the computing device 2600 includes a processing unit 2610, a network interface 2620, a computer-readable medium drive 2630, an input / output device interface 2640, a display 2650, and an input device 2660, all of which can communicate with each other via a communication bus. The network interface 2620 can provide a connection to one or more networks or computing systems. Therefore, the processing unit 2610 can receive information and instructions from other computing systems or services via the network. The processing unit 2610 can also communicate with the memory 2670, and further provide output information to the optional display 2650 via the input / output device interface 2640. For example, analysis software (e.g., data analysis software or program, such as a program) stored as executable instructions in the non-transient memory of the analysis system ) can display flow cytometry event data to a user. The input / output device interface 2640 can also accept input from an optional input device 2660, such as a keyboard, mouse, digital pen, microphone, touch screen, gesture recognition system, voice recognition system, game controller, accelerometer, gyroscope, or other input device. Data can also be stored in the data storage 2690.

[0181] Methods for analyzing samples

[0182] Aspects of the invention also include methods of analyzing a sample. Methods of interest include (a) introducing a sample into an instrument including a fluid supply system, and (b) supplying a working fluid to the instrument to analyze the sample. A fluid supply system for these methods is as described above, comprising: a working fluid line configured to deliver a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a working fluid reservoir comprising the working fluid; a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end and comprising a primary flush fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a primary flush fluid reservoir comprising the primary flush fluid; a secondary fluid line configured to deliver a secondary fluid therethrough from a proximal end to a distal end and comprising a secondary fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a secondary fluid reservoir comprising the secondary fluid; a flow control circuit fluidically connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and a single fluid connection configured to be fluidically coupled to the flow control circuit, the working fluid line, and a fluid assembly of the instrument.

[0183] In some cases, the sample analyzed in the present method is a biological sample. The term "biological sample" refers to a subset of a whole organism, plant, fungus or animal tissue, cell or component in its conventional sense, which in some cases can be found in blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, amniotic fluid, amniotic cord blood, urine, vaginal fluid and semen. Therefore, "biological sample" refers to both natural organisms or their tissue subsets, and also refers to homogenates, lysates or extracts prepared from organisms or their tissue subsets, including but not limited to, for example, plasma, serum, cerebrospinal fluid, lymph, skin sections, respiratory tract, gastrointestinal tract, cardiovascular and urogenital tract, tears, saliva, milk, blood cells, tumors, organs. Biological samples can be any type of organic tissue, including healthy and diseased tissues (e.g., cancerous, malignant, necrotic, etc.). In certain embodiments, the biological sample is a liquid sample, such as blood or a derivative thereof, such as plasma, tears, urine, semen, etc., wherein in certain cases, the sample is a blood sample, including whole blood, such as blood obtained from a venipuncture or fingertip prick (wherein the blood may or may not be mixed with any reagents, such as preservatives, anticoagulants, etc., prior to assay).

[0184] In certain embodiments, the sample source is "mammal", which terms are widely used to describe organisms in the mammal class, including Carnivora (e.g., dogs and cats), Rodentia (e.g., mice, guinea pigs and rats), and primates (e.g., humans, chimpanzees and monkeys). In some cases, the subject is human. These methods can be applied to samples obtained from human subjects of both sexes and any developmental stage (i.e., newborns, infants, teenagers, adolescents, adults), wherein in certain embodiments, the human subjects are teenagers, adolescents or adults. Although the present invention can be applied to samples from human subjects, it should be understood that these methods can also be performed on samples from other animal subjects (i.e., "non-human subjects"), such as, but not limited to, birds, mice, rats, dogs, cats, livestock and horses.

[0185] Target cells can be targeted to characterize them according to a variety of parameters, such as having phenotypic characteristics identified by attaching specific fluorescent labels to target cells. In some embodiments, the system is configured to deflect analyzed droplets determined to contain target cells. This method can be used to characterize a variety of cells. Target cells of interest include, but are not limited to, stem cells, T cells, dendritic cells, B cells, granulocytes, leukemia cells, lymphoma cells, viral cells (e.g., HIV cells), NK cells, macrophages, monocytes, fibroblasts, epithelial cells, endothelial cells, and erythrocytes. Target cells of interest include cells with convenient cell surface markers or antigens, which can be captured or labeled by convenient affinity agents or their conjugates. For example, the target cell may include cell surface antigens such as CD11b, CD123, CD14, CD15, CD16, CD19, CD193, CD2, CD25, CD27, CD3, CD335, CD36, CD4, CD43, CD45RO, CD56, CD61, CD7, CD8, CD34, CD1c, CD23, CD304, CD235a, T cell receptor alpha receptor, T cell receptor gamma receptor, CD253, CD95, CD20, CD105, CD117, CD120b, Notch4, Lgr5 (N-terminal), SSEA-3, TRA-1-60 antigen, disialoganglioside GD2, and CD71. In some embodiments, the target cell is selected from cells containing HIV, Treg cells, antigen-specific T cell populations, tumor cells, or hematopoietic progenitor cells (CD34+) from whole blood, bone marrow, or umbilical cord blood.

[0186] The method of interest may also include the use of microparticles in research, laboratory testing, or treatment. In some embodiments, the subject method includes obtaining prepared individual cells from a target fluid or tissue biological sample. For example, the subject method includes obtaining cells from a fluid or tissue sample for use as a research or diagnostic sample for a disease such as cancer. Similarly, the subject method includes obtaining cells from a fluid or tissue sample for treatment. A cell therapy protocol is a protocol in which a living cell material including, for example, cells and tissues can be prepared and introduced into a subject as a treatment method. Diseases that can be treated by administering flow cytometry sorted samples include, but are not limited to, blood diseases, immune system diseases, organ damage, and the like.

[0187] Typical cell therapy protocols may include the following steps: sample collection, cell separation, genetic modification, culture and in vitro amplification, cell harvesting, sample volume reduction and cleaning, biological preservation, storage and introduction of cells into the subject. The protocol can start with collecting living cells and tissues from the source tissue of the subject to produce samples of cells and / or tissues. The sample can be collected by any suitable procedure, including, for example, applying a cell mobilization agent to the subject, extracting blood from the subject, removing bone marrow from the subject, etc. After collecting the sample, cell enrichment can be performed by several methods, including, for example, centrifugation-based methods, filter-based methods, elutriation, magnetic separation methods, fluorescence-activated cell sorting (FACS), etc. In some cases, the enriched cells can be genetically modified by any convenient method, such as nuclease-mediated gene editing. Genetically modified cells can be cultured, activated and amplified in vitro. In some cases, the cells are preserved (e.g., cryopreserved) and stored for future use, wherein the cells are thawed and then applied to the patient, for example, the cells can be injected into the patient.

[0188] How to assemble the instrument

[0189] Aspects of the invention also include methods of assembling an instrument.Methods of interest include operably connecting a fluid supply system of the invention to a fluidic component (eg, a flow cell) of an instrument (eg, a flow cytometer). The fluid supply system for the method is as described above and includes: a working fluid line configured to deliver a working fluid therethrough from a proximal end to a distal end and including a working fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a working fluid reservoir containing the working fluid; a primary flushing fluid line configured to deliver a primary flushing fluid therethrough from a proximal end to a distal end and including a primary flushing fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a primary flushing fluid reservoir containing the primary flushing fluid; a secondary fluid line configured to deliver a secondary fluid therethrough from a proximal end to a distal end and including a secondary fluid reservoir connector located at the proximal end and configured to be fluidically coupled to a secondary fluid reservoir containing the secondary fluid; a flow control circuit fluidically connected to the distal end of the primary flushing fluid line and the distal end of the secondary fluid line and configured to selectively allow the primary flushing fluid and the secondary fluid to pass through an outlet; and a single fluid connection configured to be fluidically coupled to the flow control circuit, the working fluid line, and a fluid assembly of the instrument.

[0190] In some embodiments, the method includes fluidly coupling a working fluid reservoir to a working fluid line. In some cases, the method also includes operably connecting a pressurized air source to the working fluid reservoir. In some such cases, the method includes operably connecting a pressure regulator to the working fluid reservoir and the pressurized air source. In some cases, the method includes fluidly coupling a main flush fluid reservoir to a main flush fluid line. In selected embodiments, the method includes fluidly coupling a secondary fluid reservoir to a secondary fluid line. The method may also include fluidly coupling a plurality of secondary fluid lines to a plurality of secondary fluid reservoirs. When the flow control circuit of the fluid supply system includes a valve group, the method according to some embodiments includes fluidly coupling the distal end of the main flush fluid line to a valve in the valve group that is farthest from the outlet of the flow control circuit. In some cases, the method includes operably connecting a vacuum source to a fluid component of the instrument.

[0191] Practicality

[0192] The flow-type microparticle analyzer with a fluid supply system disclosed herein and the method of using the same can be used for a variety of different applications, including, for example, research and clinical applications. The flow-type microparticle analyzer can be used in applications where it is necessary to analyze and / or analyze and sort microparticle components in a sample (e.g., a biological sample). The fluid supply system disclosed herein can be incorporated into any suitable flow-type microparticle system (e.g., a flow cytometer system). The fluid supply system can be used in any suitable flow-type microparticle analyzer application, wherein a cleaning fluid flows through the flow-type microparticle analyzer to clean the fluid management system. The fluid supply system can be used in applications where a specific fluid path volume in the fluid management system cannot be entered and the system needs to be 100% thoroughly cleaned. The fluid management system (e.g., having a waste management control with a single fluid moving device) can be used in applications where it is necessary to reduce the cost and complexity of the production and operation of the flow-type microparticle analyzer. The fluid management system (e.g., the hybrid system described herein) can also be further used in applications where it is necessary to increase the flexibility of microparticle analysis. Suitable applications include applications where it is necessary to switch between (a) analysis and (b) analysis and sorting modes of the flow-type microparticle analyzer. Such applications may include clinical or research applications where the flexibility to perform (a) microparticle analysis or (b) microparticle analysis and sorting using a single flow cytometer system in the same laboratory or clinical setting is desirable, for example, cost-effectively.

[0193] The fluid supply system described herein can be used in a variety of different flow-type particle analyzers, such as flow cytometers. Suitable flow cytometer systems that can use the subject fluid management system include, but are not limited to, U.S. Patent Nos. 9,952,076, 9,933,341, 9,726,527, 9,453,789, 9,200,334, 9,097,640, 9,095,494, 9,092,034, 8,975,595, 8,753,573, 8,233,146, 8,140, The systems described in 300, 7,544,326, 7,201,875, 7,129,505, 6,821,740, 6,813,017, 6,809,804, 6,372,506, 5,700,692, 5,643,796, 5,627,040, 5,620,842, and 5,602,039, the disclosures of which are incorporated herein by reference in their entireties. In some cases, flow cytometry systems of interest include BD Biosciences FACSCantoTMII flow cytometer, BD AccuriTM flow cytometer, BD Biosciences FACSCelestaTM flow cytometer, BD Biosciences FACSLyricTM flow cytometer, BD Biosciences FACSVerseTM flow cytometer, BD Biosciences FACSymphonyTM flow cytometer, BD Biosciences LSRFortessaTM flow cytometer, and BD Biosciences LSRFortessTMX-20 flow cytometer, among others.

[0194] Other suitable flow cytometry systems include, but are not limited to, U.S. Pat. Nos. 3,960,449, 4,347,935, 4,667,830, 4,704,891, 4,770,992, 5,030,002, 5,040,890, 5,047,321, 5,245,318, 5,317,162, 5,464,581, 5,483,469, 5,602,039, 5,620,842, 5,627,040, 5,643,79 6, 5,700,692, 6,372,506, 6,809,804, 6,813,017, 6,821,740, 7,129,505, 7,201,875, 7,544,326, 8,140,300, 8,233,146, 8,753,573, 8,975,595, 9,092,034, 9,095,494, and 9,097,640, the disclosures of which are incorporated herein by reference in their entireties. In some cases, the sorting flow cytometer is a Becton Dickinson cell sorter, such as the BD Biosciences Influxe cell sorter, the BD Biosciences FACSAriac III and BD FACSAriac Fusion cell sorters, the BD Biosciences FACSJazzc cell sorter, and the BD Biosciences FACSMelody TM Cell sorter, etc.

[0195] Notwithstanding the appended claims, the present disclosure is also defined by the following terms:

[0196] 1. A fluid supply system, comprising:

[0197] a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and including a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid;

[0198] a primary flush fluid line configured to deliver a primary flush fluid therethrough from the proximal end to the distal end and including a primary flush fluid reservoir connector at the proximal end configured to fluidly couple to a primary flush fluid reservoir containing the primary flush fluid;

[0199] a secondary fluid line configured to deliver a secondary fluid therethrough from a proximal end to a distal end and including a secondary fluid reservoir connector at the proximal end, the secondary fluid reservoir connector configured to fluidly couple to a secondary fluid reservoir containing the secondary fluid;

[0200] a flow control circuit fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow primary flush fluid and secondary fluid to pass through the outlet; and

[0201] A single fluid connection is configured to fluidically couple to a flow control loop, a working fluid line, and a fluid component of an instrument.

[0202] 2. The fluid supply system of clause 1, wherein the single fluid connection comprises a quick connect fluid connector.

[0203] 3. The fluid supply system of clause 1, wherein the single fluid connection comprises a three-way valve.

[0204] 4. The fluid supply system according to clause 3, wherein the three-way valve has a 5 mm 3 Up to 100mm 3 The internal volume within the range.

[0205] 5. A fluid supply system according to any of the preceding clauses, further comprising a fluid supply pump operably connected to the flow control circuit and the single fluid connection.

[0206] 6. The fluid supply system according to any of the preceding clauses, further comprising a working fluid reservoir fluidly coupled to the working fluid pipeline.

[0207] 7. The fluid supply system of clause 6, wherein the working fluid is a sheath fluid.

[0208] 8. The fluid supply system of clause 6 or 7, further comprising a source of pressurized air operably connected to the working fluid reservoir.

[0209] 9. The fluid supply system of clause 8, further comprising a pressure regulator operably connected to the working fluid reservoir and the pressurized air source.

[0210] 10. A fluid supply system according to any of the preceding clauses, further comprising a working fluid supply pump operably connected to the single fluid connection.

[0211] 11. The fluid supply system according to any of the preceding clauses, further comprising a primary flush fluid reservoir fluidly coupled to the primary flush fluid line.

[0212] 12. The fluid supply system of clause 11, wherein the primary flushing fluid is an inert fluid.

[0213] 13. The fluid supply system according to any of the preceding clauses, further comprising a secondary fluid reservoir fluidly coupled to the secondary fluid line.

[0214] 14. The fluid supply system of clause 13, wherein the secondary fluid is selected from a cleaning fluid and a reagent.

[0215] 15. The fluid supply system according to any of the preceding clauses, wherein the fluid supply system comprises a plurality of secondary fluid lines.

[0216] 16. The fluid supply system of clause 15, wherein the number of the plurality of secondary fluid lines ranges from 1 to 10.

[0217] 17. A fluid supply system according to any of the preceding clauses, wherein the flow control circuit comprises a valve assembly.

[0218] 18. The fluid supply system of clause 17, wherein the flow control circuit comprises a two-way valve set.

[0219] 19. The fluid supply system of clause 18, wherein the two-way valve is an independent valve.

[0220] 20. The fluid supply system of clause 18, wherein the two-way valve is mounted on the manifold.

[0221] 21. The fluid supply system of clause 17, wherein the flow control circuit comprises a three-way valve set.

[0222] 22. The fluid supply system of clause 21, wherein the three-way valve is an independent valve.

[0223] 23. The fluid supply system of clause 21, wherein the three-way valve is mounted on the manifold.

[0224] 24. The fluid supply system of any of clauses 17 to 23, wherein the distal end of the main flush fluid line is fluidly coupled to a valve in the valve group that is furthest from the outlet of the flow control circuit.

[0225] 25. A fluid supply system according to any of clauses 1 to 16, wherein the flow control circuit comprises a multi-port selector valve.

[0226] 26. The fluid supply system according to clause 25, wherein the multi-port selector valve has a range from 5 mm 3 Up to 1000mm 3 internal volume.

[0227] 27. A fluid supply system according to any of the preceding clauses, further comprising an instrument.

[0228] 28. A fluid supply system according to clause 27, wherein the instrument is a flow cytometer.

[0229] 29. A fluid supply system according to clause 28, wherein the fluidic component of the instrument is a flow cell.

[0230] 30. The fluid supply system of any of clauses 27 to 29, further comprising a vacuum source operably connected to the fluidic component of the instrument.

[0231] 31. A method for analyzing a sample, the method comprising:

[0232] (a) introducing a sample into an instrument comprising a fluid supply system, the fluid supply system comprising:

[0233] a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid;

[0234] a primary flush fluid line configured to deliver a primary flush fluid therethrough from the proximal end to the distal end and comprising a primary flush fluid reservoir connector at the proximal end configured to fluidly couple to a primary flush fluid reservoir containing the primary flush fluid;

[0235] a secondary fluid line that: is configured to deliver a secondary fluid therethrough from a proximal end to a distal end and includes a secondary fluid reservoir connector at the proximal end, the secondary fluid reservoir connector being configured to fluidly couple to a secondary fluid reservoir containing the secondary fluid;

[0236] a flow control circuit that is: fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow primary flush fluid and secondary fluid to pass through the outlet; and

[0237] a single fluid connection that: is configured to fluidically couple to a flow control loop, a working fluid line, and a fluid component of an instrument; and

[0238] (b) Supplying working fluid to the instrument to analyze the sample.

[0239] 32. The method of clause 31, wherein the single fluid connection comprises a quick connect fluid connector.

[0240] 33. The method of clause 31, wherein the single fluid connection comprises a three-way valve.

[0241] 34. The method according to clause 33, wherein the three-way valve has a range from 5 mm 3 Up to 100mm 3 internal volume.

[0242] 35. The method of any of clauses 31 to 34, wherein the fluid supply subsystem further comprises a fluid supply pump operably connected to the flow control circuit and the single fluid connection.

[0243] 36. The method of any of clauses 31 to 35, wherein the fluid supply system further comprises a working fluid reservoir fluidly coupled to the working fluid line.

[0244] 37. The method of clause 36, wherein the working fluid is a sheath fluid.

[0245] 38. A method according to clause 36 or 37, wherein the fluid supply system further comprises a source of pressurized air operably connected to the working fluid reservoir.

[0246] 39. The method of clause 38, wherein the fluid control system further comprises a pressure regulator operably connected to the working fluid reservoir and the pressurized air source.

[0247] 40. A method according to any of clauses 31 to 39, wherein the fluid supply system further comprises a working fluid supply pump operably connected to the single fluid connection.

[0248] 41. The method of any of clauses 31 to 40, wherein the fluid supply system further comprises a primary flush fluid reservoir fluidly coupled to the primary flush fluid line.

[0249] 42. The method of clause 41, wherein the primary flushing fluid is an inert fluid.

[0250] 43. The method of any of clauses 31 to 42, wherein the fluid supply system further comprises a secondary fluid reservoir fluidly coupled to the secondary fluid line.

[0251] 44. The method of clause 43, wherein the secondary fluid is selected from a cleaning fluid and a reagent.

[0252] 45. A method according to any of clauses 31 to 44, wherein the fluid supply system comprises a plurality of secondary fluid lines.

[0253] 46. ​​The method of clause 45, wherein the number of the plurality of secondary fluid lines ranges from 1 to 10.

[0254] 47. A method according to any of clauses 31 to 46, wherein the flow control circuit comprises a valve assembly.

[0255] 48. The method of clause 47, wherein the flow control circuit comprises a two-way valve set.

[0256] 49. The method of clause 48, wherein the two-way valve is an independent valve.

[0257] 50. The method of clause 48, wherein the two-way valve is mounted on the manifold.

[0258] 51. The method of clause 47, wherein the flow control circuit comprises a three-way valve set.

[0259] 52. The method of clause 51, wherein the three-way valve is an independent valve.

[0260] 53. The method of clause 51, wherein the three-way valve is mounted on the manifold.

[0261] 54. The method of any of clauses 47 to 53, wherein the distal end of the main flush fluid line is fluidly coupled to a valve in the valve group that is farthest from an outlet of the flow control circuit.

[0262] 55. A method according to any of clauses 31 to 46, wherein the flow control circuit comprises a multi-port selector valve.

[0263] 56. The method according to clause 55, wherein the multi-port selector valve has a range from 5 mm 3 Up to 1000mm 3 internal volume.

[0264] 57. A method according to any one of clauses 31 to 56, wherein the instrument is a flow cytometer.

[0265] 58. A method according to clause 57, wherein the fluidic component of the instrument is a flow cell.

[0266] 59. A method according to any of clauses 57 to 58, wherein the fluid supply system comprises a vacuum source operably connected to the fluidic component of the instrument.

[0267] 60. A method according to any one of clauses 31 to 59, wherein the sample is a biological sample.

[0268] 61. A method of assembling an instrument, the method comprising operably connecting a fluid supply system to a fluid component of the instrument, the fluid supply system comprising:

[0269] a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid;

[0270] a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end and comprising a primary flush fluid reservoir connector at said proximal end, said primary flush fluid reservoir connector configured to fluidly couple to a primary flush fluid reservoir, said primary flush fluid reservoir containing said primary flush fluid;

[0271] a secondary fluid line that: is configured to deliver a secondary fluid therethrough from a proximal end to a distal end and includes a secondary fluid reservoir connector at said proximal end, said secondary fluid reservoir connector being configured to fluidly couple to a secondary fluid reservoir, said secondary fluid reservoir containing said secondary fluid;

[0272] a flow control circuit that is: fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and

[0273] A single fluid connection is configured to fluidically couple to the flow control circuit, the working fluid line, and a fluid component of an instrument.

[0274] 62. The method of clause 61, wherein the single fluid connection comprises a quick connect fluid connector.

[0275] 63. The method of clause 61, wherein the single fluid connection comprises a three-way valve.

[0276] 64. The method according to clause 63, wherein the three-way valve has a range from 5 mm 3 Up to 100mm 3 internal volume.

[0277] 65. The method of any of clauses 61 to 64, wherein the fluid supply system further comprises a fluid supply pump operably connected to the flow control circuit and the single fluid connection.

[0278] 66. The method of any of clauses 61 to 65, further comprising fluidly coupling a working fluid reservoir to the working fluid line.

[0279] 67. The method of clause 66, wherein the working fluid is a sheath fluid.

[0280] 68. The method of clause 66 or 67, further comprising operably connecting a source of pressurized air to the working fluid reservoir.

[0281] 69. The method of clause 68, further comprising operably connecting a pressure regulator to the working fluid storage tank and the pressurized air source.

[0282] 70. A method according to any of clauses 61 to 69, wherein the fluid supply system further comprises a working fluid supply pump operably connected to the single fluid connection.

[0283] 71. The method of any of clauses 61 to 70, further comprising fluidly coupling a primary flush fluid reservoir to the primary flush fluid line.

[0284] 72. The method of clause 71, wherein the primary flushing fluid is an inert fluid.

[0285] 73. The method of any of clauses 61 to 72, further comprising fluidly coupling a secondary fluid reservoir to the secondary fluid line.

[0286] 74. The method of clause 73, wherein the secondary fluid is selected from a cleaning fluid and a reagent.

[0287] 75. A method according to any of clauses 61 to 74, wherein the fluid supply system comprises a plurality of secondary fluid lines.

[0288] 76. The method of clause 75, wherein the number of the plurality of secondary fluid lines ranges from 1 to 10.

[0289] 77. A method according to any of clauses 61 to 76, wherein the flow control circuit comprises a valve assembly.

[0290] 78. The method of clause 77, wherein the flow control circuit comprises a two-way valve set.

[0291] 79. A method according to clause 78, wherein the two-way valve is an independent valve.

[0292] 80. The method of clause 78, wherein the two-way valve is mounted on the manifold.

[0293] 81. The method of clause 77, wherein the flow control circuit comprises a three-way valve set.

[0294] 82. A method according to clause 81, wherein the three-way valve is an independent valve.

[0295] 83. The method of clause 81, wherein the three-way valve is mounted on the manifold.

[0296] 84. The method of any of clauses 77 to 83, further comprising fluidly coupling a distal end of the primary flush fluid line to a valve in the valve group that is farthest from an outlet of the flow control circuit.

[0297] 85. A method according to any of clauses 61 to 76, wherein the flow control circuit comprises a multi-port selector valve.

[0298] 86. The method of clause 85, wherein the multi-port selector valve has a range from 5 mm 3 Up to 1000mm 3 internal volume.

[0299] 87. A method according to any one of clauses 61 to 86, wherein the instrument is a flow cytometer.

[0300] 88. A method according to clause 87, wherein the fluid component of the instrument is a flow cell.

[0301] 89. The method of any one of clauses 87 to 88, further comprising operably connecting a vacuum source to a fluid component of the instrument.

[0302] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to those skilled in the art that, based on the teachings of the present invention, changes and modifications may be readily made thereto without departing from the spirit or scope of the appended claims.

[0303] Therefore, the foregoing only illustrates the principles of the present invention. It is understood that those skilled in the art will be able to design various arrangements that, although not explicitly described or shown herein, embody the principles of the present invention and are included in its spirit and scope. In addition, all examples and conditional language described herein are primarily intended to help readers understand the principles of the present invention and the concepts contributed by the inventors to promote technological progress, and should be interpreted as not being limited to these specifically described examples and conditions. In addition, all statements described herein, including the principles, aspects and embodiments of the present invention and their specific examples, are intended to cover their structural and functional equivalents. In addition, such equivalents are intended to include currently known equivalents and equivalents developed in the future, that is, regardless of the structure, any element developed to perform the same function is included. In addition, nothing disclosed herein is intended to be dedicated to the public, regardless of whether such disclosure is explicitly recorded in the claims.

[0304] Therefore, the scope of the present invention is not intended to be limited to the exemplary embodiments shown and described herein. Instead, the scope and spirit of the present invention are embodied by the appended claims. In the claims, only when the precise phrase "device for..." or the precise phrase "step for..." is cited at the beginning of the limitation in the claim, it is explicitly defined to refer to 35 U.S.C. §112(f) or 35 U.S.C. §112(6) as a limitation in the claim; if such precise phrases are not used in the limitation in the claim, 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is not cited.

Claims

1. A fluid supply system, comprising: a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid; a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end and comprising a primary flush fluid reservoir connector at said proximal end, said primary flush fluid reservoir connector configured to fluidly couple to a primary flush fluid reservoir, said primary flush fluid reservoir containing said primary flush fluid; a secondary fluid line that: is configured to deliver a secondary fluid therethrough from a proximal end to a distal end and includes a secondary fluid reservoir connector at said proximal end, said secondary fluid reservoir connector being configured to fluidly couple to a secondary fluid reservoir, said secondary fluid reservoir containing said secondary fluid; a flow control circuit that is: fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and A single fluid connection is configured to fluidically couple to the flow control circuit, the working fluid line, and a fluid component of an instrument.

2. The fluid supply system according to claim 1, wherein: The single fluid connection comprises a quick connect fluid connector.

3. The fluid supply system according to claim 1, wherein: The single fluid connection comprises a three-way valve.

4. The fluid supply system of any one of the preceding claims, further comprising a fluid supply pump operably connected to the flow control circuit and the single fluid connection.

5. The fluid supply system of any of the preceding claims, further comprising the working fluid reservoir, the working fluid reservoir being fluidly coupled to the working fluid line.

6. A fluid supply system according to any preceding claim, further comprising a working fluid supply pump operatively connected to the single fluid connection.

7. The fluid supply system of any of the preceding claims, further comprising the primary flush fluid reservoir, the primary flush fluid reservoir being fluidly coupled to the primary flush fluid line.

8. The fluid supply system of any of the preceding claims, further comprising the secondary fluid reservoir, the secondary fluid reservoir being fluidly coupled to the secondary fluid line.

9. A fluid supply system according to any one of the preceding claims, wherein: The fluid supply system includes a plurality of secondary fluid lines.

10. A fluid supply system according to any one of the preceding claims, wherein: The flow control circuit includes a valve group.

11. The fluid supply system according to any one of claims 1 to 11, wherein: The flow control circuit includes a multi-port selector valve.

12. The fluid supply system according to any one of the preceding claims, further comprising a flow cytometer.

13. A method for analyzing a sample, the method comprising: (a) introducing the sample into an instrument comprising a fluid supply system, the fluid supply system comprising: a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid; a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end and comprising a primary flush fluid reservoir connector at said proximal end, said primary flush fluid reservoir connector configured to fluidly couple to a primary flush fluid reservoir containing said primary flush fluid; a secondary fluid line that: is configured to deliver a secondary fluid therethrough from a proximal end to a distal end and includes a secondary fluid reservoir connector at said proximal end, said secondary fluid reservoir connector being configured to fluidly couple to a secondary fluid reservoir containing said secondary fluid; a flow control circuit that is: fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and a single fluid connection configured to fluidically couple to the flow control circuit, the working fluid line, and a fluid component of an instrument; and (b) supplying the working fluid to the instrument to analyze the sample.

14. The method according to claim 13, wherein: The flow control circuit is according to any one of claims 2 to 12.

15. A method of assembling an instrument, the method comprising operably connecting a fluid supply system to a fluid component of the instrument, the fluid supply system comprising: a working fluid line configured to convey a working fluid therethrough from a proximal end to a distal end and comprising a working fluid reservoir connector at the proximal end, the working fluid reservoir connector configured to fluidly couple to a working fluid reservoir containing the working fluid; a primary flush fluid line configured to deliver a primary flush fluid therethrough from a proximal end to a distal end and comprising a primary flush fluid reservoir connector at said proximal end, said primary flush fluid reservoir connector configured to fluidly couple to a primary flush fluid reservoir, said primary flush fluid reservoir containing said primary flush fluid; a secondary fluid line that: is configured to deliver a secondary fluid therethrough from a proximal end to a distal end and includes a secondary fluid reservoir connector at said proximal end, said secondary fluid reservoir connector being configured to fluidly couple to a secondary fluid reservoir, said secondary fluid reservoir containing said secondary fluid; a flow control circuit that is: fluidly connected to a distal end of the primary flush fluid line and a distal end of the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and A single fluid connection is configured to fluidically couple to the flow control circuit, the working fluid line, and a fluid component of an instrument.

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