Fluid supply systems having flow control circuit and single fluidic connection, and methods of use thereof

Abstract

To provide fluid supply systems configured to minimize carryover.SOLUTION: Fluid supply systems of interest include an operating 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, and configured to selectively permit passage of a primary flushing fluid and a secondary fluid through an outlet, and a single fluidic connection configured to fluidically couple to the flow control circuit, the operating fluid line, and a fluidic component of an instrument. Methods of analyzing a sample and assembling an instrument are also provided.SELECTED DRAWING: None

Description

Background Art

[0001] Flow-type particle analyzers such as flow cytometers are well-known analytical tools that enable the characterization of particles based on optical parameters such as light scattering and fluorescence, or by electrical properties such as impedance. For example, in a flow cytometer, particles such as molecules, analyte-binding beads, or individual cells in a fluid suspension pass through a detection region where the particles are typically exposed to excitation light 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 a fluorescent dye to facilitate detection. By labeling a number of different particles or components with spectrally distinct fluorescent dyes, various particles or components may be detected simultaneously. Usually, detection is performed using a number of photodetectors, with one photodetector used for each distinct dye to be detected.

Summary of the Invention

Problems to be Solved by the Invention

[0002] In some flow cytometers, sheath fluid is supplied to the flow cell by a pressure-driven fluid system, and the sample fluid and the sheath fluid pass through the flow cell at a pressure higher than the ambient pressure. The flow rate through the flow cell is changed by varying the pressure in the sheath fluid reservoir, and the ratio of the sample fluid to the sheath fluid in the hydrodynamic flow is determined by the pressures applied in the sample source and the sheath fluid reservoir and the resistance of the fluid system supplying the sample fluid and the sheath fluid. A vacuum-driven fluid system can further be used in the flow cytometer, where a vacuum pump evacuates the downstream side of the flow cell and the sample fluid and the sheath fluid remain at ambient pressure. To vary the flow rate through the flow cell, the vacuum pump evacuates it, and the ratio of the sample fluid and the sheath fluid flowing through the flow cell is determined by the ratio of the resistances due to the paths of the sample fluid system and the sheath fluid system. Fluid systems that supply a hydrodynamically focused flow of the sample fluid containing particles to the center of the flow of the sheath fluid containing no particles often utilize pressurizable tubing, connectors, and seals that need to withstand a wide range of pressure levels, particularly high and low pressures.

[0003] Pressure-based fluid systems configured to supply the flow of the sample fluid to the flow of the sheath fluid include the BD Biosciences FACSAria™ I, BD Biosciences FACSAria™ II, BD Biosciences FACSAria™ III, BD Biosciences FACSCanto™ flow cytometers (described in U.S. Patent No. 6,767,188), and the Bio-Rad S3e™ (described in U.S. Patent No. 9,551,637). These systems may be configured to switch fluids so that various fluids can be supplied to the flow cell at various times as desired. Often, switching of the fluids involves a manual process where the user reconnects the fluid lines to different fluid sources.

[0004] While the pressure-based fluid system as shown in U.S. Patent No. 6,767,188 improves fluid engineering technology, the inventor recognizes that further improvements can be made. In particular, it has been discovered that the problem with conventional fluid supply mechanisms is to prevent carry-over between fluids (i.e., the first fluid remaining in the fluid line used to carry the second fluid). In certain applications, for example, when the cleaning fluid can affect the purity of the fluid, when the cleaning fluid can affect the biological sample of the device, when multiple cleaning fluids or reagents are required but direct mixing of the fluids can cause undesirable chemical reactions, it is important that the carry-over between multiple fluids be very low. This means that the upward line can (1) potentially be a source of contamination for the device or (2) require manual removal and cleaning by the user. Since the upward line is not completely swept, it does not guarantee low fluid carry-over without manual intervention by the user. Therefore, a fluid supply system that is configured to be completely swept and have minimal carry-over is desirable. The systems and methods of the present invention meet this need.

Means for Solving the Problems

[0005] Aspects of the present invention include a fluid supply system. The subject system includes an actuation fluid line having a proximal end with an actuation fluid reservoir connector configured to fluidly couple to an actuation fluid reservoir containing actuation fluid (e.g., sheath fluid) and configured to carry the actuation fluid from the proximal end to the distal end; a primary flush fluid line having a proximal end with a primary flush fluid reservoir connector configured to fluidly couple to a primary flush fluid reservoir containing primary flush fluid (e.g., inert fluid) and configured to carry the primary flush fluid from the proximal end to the distal end; a secondary fluid line having a proximal end with a secondary fluid reservoir connector configured to fluidly couple to a secondary fluid reservoir containing secondary fluid (e.g., cleaning solution, reagent, etc.) and configured to carry the secondary fluid from the proximal end to the distal end; a flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet; and one fluid connection configured to fluidly couple to the flow control circuit, the actuation fluid line, and the fluid components of the device. The one fluid connection may have, for example, a quick connect fluid connector or a three-way valve. The system of interest further includes a fluid supply pump operably connected to the flow control circuit and the one fluid connection. In embodiments, there is further provided an actuation fluid reservoir fluidly coupled to the actuation fluid line and / or a pressure regulator operably connected to the actuation fluid reservoir and a pressurized air source. In some cases, the system includes an actuation fluid supply pump operably connected to the one fluid connection. In a particular variation, the system includes a primary flush fluid reservoir fluidly coupled to the primary flush fluid line. The system of some embodiments includes a secondary fluid reservoir fluidly 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 has a row of valves. The row of valves may have, for example, two-way valves or three-way valves. In some cases, the two-way valve or three-way valve is a stand-alone valve.In other cases, the two-way valve or three-way valve is attached to the manifold. In certain cases, the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is furthest from the outlet of the flow control circuit. In some embodiments, the flow control circuit has a multi-port selection valve.

[0006] In some embodiments, the system of the present invention further comprises a device. The device may have, for example, a flow cytometer. In some such cases, the fluid component of the device is a flow cell. In certain cases, the system further comprises a vacuum source operably connected to the fluid component of the device. Aspects of the present invention further include a method of analyzing a sample. In the method of interest, a sample is introduced into a device comprising the fluid supply system of the present invention, and an operating fluid is supplied to the device to analyze the sample. Aspects of the present invention further include a method of assembling a device. In such a method, the fluid supply system of the present invention is operably connected to the fluid component of the device.

Brief Description of the Drawings

[0007] The present invention can be best understood from the following detailed description when read in conjunction with the accompanying drawings. The drawings include the following figures.

[0008]

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Embodiments for Carrying Out the Invention

[0009] A fluid supply system is provided. The fluid supply system of interest includes a working fluid line, a primary flush fluid line, a secondary fluid line, a flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively enable the primary flush fluid and the secondary fluid to pass through an outlet, and one fluid connection configured to fluidly couple the flow control circuit, the working fluid line, and the fluid components of the device. A method of analyzing a sample and a method of assembling a device are further provided.

[0010] Before the present invention is described in more detail, it is to be understood that the invention is not limited to the specific embodiments described, as the invention itself may of course vary. Since the scope of the present invention is defined only by the appended claims, it is to be further understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0011] Where a range of values is provided, each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limits of that range, and any other stated value or intervening value within the stated range, is to be understood as being included in the invention, unless the context clearly dictates otherwise. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are further included in the invention, subject to any specifically excluded limit within the stated range. Where the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the invention.

[0012] In this specification, a range is presented with the term "about" preceding a numerical value. The term "about" is used in this specification to literally support the exact number that the term precedes, and a number that is close to or approximates the number that the term precedes. When determining whether a number is close to or approximates a specifically recited number, the number that is not described as being close or approximate may be a number that gives a substantial equivalent of the specifically recited number in the context in which the specifically recited number is presented.

[0013] Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be further used in the practice or testing of the present invention, representative and exemplary methods and materials are described.

[0014] All publications and patents cited herein are incorporated herein by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference, and are incorporated herein by reference to disclose and describe the methods and / or materials related to the citation of the publications. Any citation of a publication is for its disclosure prior to the filing date, and the present invention should not be construed as admitting that the present invention has no right to precedence over such a publication on the ground of a prior invention. Further, the dates of the publications provided may be different from the actual publication dates and may need to be individually confirmed.

[0015] As used in this specification and the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should further be noted that the claims may be drafted to exclude any optional element. Accordingly, this description is intended to serve as a preamble for the use of exclusive terms such as "solely", "only", etc. in the recitation of claim elements, or for the use of "negative" limitations.

[0016] As will be apparent to those skilled in the art upon reading this disclosure, the individual embodiments described and illustrated herein each have separate components and features, and the separate components and features may be readily separated from, or readily combined with, any of the features of other embodiments without departing from the scope or spirit of the invention. All of the described methods may be performed in the order of the events described, or in any other logically possible order.

[0017] The systems and methods are described or are being described for grammatical fluidity with functional explanations, but the claims should not necessarily be construed as limited in any way by a "means" or "step" construction, unless expressly set forth under 35 U.S.C. § 112, and should be given the meaning ascribed by, and the full scope of, the definitions given by the claims under the doctrine of equivalents, and where the claims are expressly set forth under 35 U.S.C. § 112, it should be clearly understood that full statutory equivalents under 35 U.S.C. § 112 should be given.

[0018] Fluid supply system Aspects of the present invention include a fluid supply system. The fluid supply system of the present invention may be configured to selectively supply fluid, for example, to fluid components of a device. "Selectively" supplying fluid means that the fluid supply system is configured to supply one fluid and exclude other fluids. The selectively supplied fluid may contain a minimal amount of other fluids (or in some cases no other fluids). In other words, the fluid supply system is configured to reduce or eliminate carryover between fluids during operation. Reducing or eliminating carryover may include (1) reducing or removing the amount of fluid that is carried over, and / or (2) reducing or removing the number of occurrences of carryover. For example, the fluid supply system of the present invention may be configured to reduce carryover by 80% or more, for example 85% or more, for example 90% or more, for example 95% or more, for example 97% or more, for example 98% or more, for example 99% or more, for example 100%. In other words, the fluid supply system may be configured to enable 100% flushing. The fluid supply system may be configured such that a working fluid (e.g., sheath fluid) and one or more primary flush fluids or secondary fluids flow through the same one or more fluid paths.

[0019] The fluid supply system of the present disclosure may be configured to supply fluid to a fluid component of a device, such as a flow cell. The fluid supply system may be fluidly coupled to the input of the fluid component (such as a flow cell). The fluid supply system may be configured to selectively supply to the input of the fluid component (a) a working fluid, or (b) one or more of a primary flush fluid and one or more secondary fluids. "Selectively supply" means that the fluid supply system can supply (a) the working fluid to the input of the flow cell, (b) one or more of the primary flush fluid and one or more secondary fluids to the input of the flow cell, but only one of (a) the working fluid and (b) the primary flush fluid and one or more of the secondary fluids can be supplied at a time. For example, the fluid supply system can supply (a) the working fluid to the input of the fluid component, (b) one or more of the primary flush fluids or secondary fluids to the input of the fluid component, but may be configured such that (a) in some cases the working fluid is supplied, or (b) in other cases one or more of the primary flush fluids or secondary fluids are supplied. The fluid supply system may be configured to selectively and fluidly couple (a) a working fluid reservoir, or (b) one or more primary flush fluid reservoirs and / or secondary fluid reservoirs to the input of the fluid component. "Selectively and fluidly couple" means that the fluid supply system can fluidly couple (a) the working fluid reservoir to the input of the fluid component, (b) one or more of the primary flush fluid reservoirs and / or secondary fluid reservoirs to the input of the fluid component, but only one of (a) the working fluid reservoir and (b) one or more of the primary flush fluid reservoirs and / or secondary fluid reservoirs can be fluidly coupled to the input of the fluid component at a time. In some cases, the fluid coupling may be automatic / automated such that no user intervention is required. Such automation may be achieved, for example, using a processor-controlled valve.

[0020] The fluid supply system of the present invention includes a working fluid line configured to carry a working fluid from a proximal end to a distal end through the working fluid. The working fluid line may be composed of, for example, tubing or pipes and may be manufactured from any convenient material. The materials of interest include, but are not limited to, polymeric materials such as plastic materials. In some cases, the working fluid line is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), Tygon, fluorinated ethylene propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), polyethylene (PE), etc. and combinations thereof. The working fluid line may be characterized by any convenient inner diameter (i.e., the diameter inside the working fluid line), outer diameter (i.e., the diameter outside the working fluid line), wall thickness, and length. In some cases, the inner diameter of the working fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. In some cases, the outer diameter of the working fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. The working fluid line of the present invention further has a working fluid reservoir connector at the proximal end configured to be fluidly coupled to a working fluid reservoir containing the working fluid. "Fluidly coupled" means coupling in a manner sufficient to allow the passage of fluid. Any suitable connector may be included. Exemplary connectors include, for example, quick disconnect connectors, threaded connectors, luer connectors, multiport connectors, tri-clamp fittings, and puncture seal sterilization fittings. Suitable quick disconnect connectors include, but are not limited to, snap-type (ball latch) connectors, bayonet connectors, threaded connectors, non-latch connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, drive break connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0021] The actuating fluid reservoir may be any suitable reservoir or container (e.g., having a rigid or flexible wall) for holding the actuating fluid. The actuating fluid can be various fluids, and in some cases, it is the sheath fluid. The sheath fluid may include, for example, phosphate buffered saline (PBS), HEPES buffered saline, sodium fluoride (NAF), etc. and combinations thereof. The volume of the actuating fluid reservoir may be in the range of 1 L to 100 L. For example, the volume of the container may be in the range of 1 L to 90 L, 1 L to 80 L, 1 L to 70 L, 1 L to 60 L, 1 L to 50 L, 1 L to 40 L, 1 L to 30 L, 1 L to 20 L, or 1 L to 10 L. In some cases, the actuating fluid reservoir connector is configured to be detachably connected to the actuating fluid reservoir. "Detachably connected" means that the connector can be attached to a second connector, e.g., a mating connector, as desired, and then removed from the second connector, e.g., the mating connector, at a certain point. In some embodiments, the actuating fluid reservoir connector is aseptically connected to the actuating fluid reservoir. "Aseptic" means that the connection part is isolated from living bacteria or other microorganisms present in the surrounding environment. If desired, the actuating fluid reservoir, e.g., the sheath fluid container, may be individually cleaned using, for example, an autoclave cycle, etc.

[0022] The subject fluid supply system further comprises a primary flush fluid line configured to carry a primary flush fluid from a proximal end to a distal end. The primary flush fluid line may be composed of, for example, tubing or pipes and may be manufactured from any convenient material. Materials of interest include, but are not limited to, polymeric materials such as plastic materials. In some cases, the working fluid line is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), Tygon, fluorinated ethylene propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), polyethylene (PE), etc. and combinations thereof. The primary flush fluid line may be characterized by any convenient inner diameter (i.e., the diameter inside the primary flush fluid line), outer diameter (i.e., the diameter outside the primary flush fluid line), wall thickness, and length. In some cases, the inner diameter of the primary flush fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. In some cases, the outer diameter of the primary flush fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. The primary flush fluid line of the present invention further has a primary flush fluid reservoir connector at the proximal end configured to be fluidly coupled to a primary flush fluid reservoir containing the primary flush fluid. Any suitable connector may be included. Exemplary connectors include, for example, quick disconnect connectors, threaded connectors, luer connectors, multi-port connectors, tri-clamp fittings, and puncture seal sterilization fittings. Suitable quick disconnect connectors include, but are not limited to, snap type (ball latch) connectors, bayonet connectors, threaded connectors, non-latch connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, drive break connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0023] The primary flush fluid reservoir may be any suitable reservoir or container (e.g., having a rigid or flexible wall) for holding the primary flush fluid. The primary flush fluid may vary. In some cases, the primary flush fluid is an inert fluid. An "inert" fluid means a non-reactive fluid. In some embodiments, when the primary flush fluid is used for flushing the fluid supply system, the primary flush fluid may be miscible with other fluids used in the present invention (e.g., the working fluid, one or more secondary fluids) such that any remaining working fluid or secondary fluid is carried from the fluid supply system by the primary flush fluid. Exemplary primary flush fluids include, for example, deionized water or physiological saline (such as phosphate buffered saline, HEPES buffered saline, sodium fluoride solution, etc.). The volume of the primary flush fluid reservoir may be in the range of 1 L to 100 L. For example, the volume of the container may be in the range of 1 L to 90 L, 1 L to 80 L, 1 L to 70 L, 1 L to 60 L, 1 L to 50 L, 1 L to 40 L, 1 L to 30 L, 1 L to 20 L, or 1 L to 10 L. In some cases, the primary flush fluid reservoir connector is configured to be removably connected to the primary flush fluid reservoir. In some embodiments, the primary flush fluid reservoir connector makes a sterile connection to the primary flush fluid reservoir. If desired, the primary flush fluid reservoir may be individually cleaned using, for example, an autoclave cycle, etc.

[0024] The fluid supply system of the present invention further comprises a secondary fluid line configured to carry a secondary fluid from a proximal end to a distal end. The secondary fluid line may be composed of, for example, tubing or pipes and may be manufactured from any convenient material. Materials of interest include, but are not limited to, polymeric materials such as plastic materials. In some cases, the working fluid line is composed of polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), Tygon, fluorinated ethylene propylene (FEP), ethylene tetrafluoroethylene (ETFE), polypropylene (PP), polyethylene (PE), etc. and combinations thereof. The secondary fluid line may be characterized by 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 of the secondary fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. In some cases, the outer diameter of the secondary fluid line is in the range of 0.2 mm to 10 mm, for example, 1 mm to 5 mm. The secondary fluid line of the present invention further has a secondary fluid reservoir connector at the proximal end configured to be fluidly coupled to a secondary fluid reservoir containing the 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 fittings, and puncture seal sterilization fittings. Suitable quick disconnect connectors include, but are not limited to, snap (ball latch) connectors, bayonet connectors, threaded connectors, non-latch connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, drive break connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors.

[0025] The secondary fluid reservoir may be any suitable reservoir or container (e.g., having a rigid or flexible wall) for holding the secondary fluid. The secondary fluid may be various. The secondary fluid may include, for example, a cleaning solution, a buffer solution, a calibration solution, a setting bead solution, etc. In some cases, the secondary fluid is selected from a cleaning solution and a reagent. When the secondary fluid is a cleaning solution, it may include any fluid suitable for disinfecting the fluid supply system and / or the fluid components / devices connected to the fluid supply system. In some embodiments, the cleaning solution has bactericidal and / or antibacterial properties. When the secondary fluid is a reagent, any reagent suitable for use in a flow-type microfluidic environment may be used. The volume of the secondary fluid reservoir may be in the range of 1 L to 100 L. For example, the volume of the container may be in the range of 1 L to 90 L, 1 L to 80 L, 1 L to 70 L, 1 L to 60 L, 1 L to 50 L, 1 L to 40 L, 1 L to 30 L, 1 L to 20 L, or 1 L to 10 L. In some cases, the secondary fluid reservoir connector is configured to be removably connected to the secondary fluid reservoir. In some embodiments, the secondary fluid reservoir connector is aseptically connected to the secondary fluid reservoir. If desired, the secondary fluid reservoir may be individually cleaned using, for example, an autoclave cycle, etc.

[0026] In an embodiment of the fluid supply system, a plurality of secondary fluid lines and / or secondary fluid reservoirs are provided. In some embodiments, the number of secondary fluid lines of the fluid supply system is in the range of 2 to 15, for example, in the range of 2 to 10, for example, in the range of 2 to 5. In a variant, the fluid supply system comprises 2 or more secondary fluid lines, for example 3 or more secondary fluid lines, for example 4 or more secondary fluid lines, for example 5 or more secondary fluid lines, for example 6 or more secondary fluid lines, for example 7 or more secondary fluid lines, for example 8 or more secondary fluid lines, for example 9 or more secondary fluid lines, for example 10 or more secondary fluid lines. In some cases, each secondary fluid line of the fluid supply system is operably connected to a secondary fluid reservoir. The fluids in the secondary fluid reservoirs may be the same or different. In an example where three secondary fluid lines and secondary fluid reservoirs are provided, the first secondary fluid reservoir contains a cleaning liquid, the second secondary fluid reservoir contains a reagent, and the third secondary fluid reservoir contains a different reagent.

[0027] The system of the present invention further comprises a flow control circuit that is fluidly connected to the distal end of the primary flush fluid line and the distal end(s) of the one or more secondary fluid lines. The flow control circuit of the present invention is configured to selectively enable the primary flush fluid and the one or more secondary fluids to pass through the outlet. In other words, the flow control circuit of the present invention may be configured to allow the passage of a particular fluid while blocking the passage of other fluids. In some cases, the flow control circuit is configured to allow a single (i.e., one) fluid to pass through at a given time. In other cases, the flow control circuit is configured to allow the passage of multiple fluids while blocking the passage of other fluids. The mechanism for selectively enabling the passage of fluids may vary. In one variant, the flow control circuit has a row of valves. The number of valves in a row can be in the range of, for example, 3 to 10, for example, 4 to 6. In some cases, there are 2 or more valves in a row, for example, 3 or more valves, for example, 4 or more valves, for example, 5 or more valves, for example, 6 or more valves, for example, 7 or more valves, for example, 8 or more valves, for example, 9 or more valves, for example, 10 or more valves. In one variant, the flow control circuit has a row of two-way valves (i.e., valves having one inlet and one outlet). In some such variants, the flow control circuit has one valve for each of the primary flush fluid reservoir and the secondary fluid reservoir. In other embodiments, the flow control circuit has a row of three-way valves. In some such embodiments, the primary flush fluid line shares a connection with the secondary fluid line via a three-way valve. Additional three-way valves may be added for each additional secondary fluid. In some cases, the valves (e.g., two-way valves, three-way valves) are stand-alone valves. By "stand-alone" valves, it means that each valve in a row is a separate component and is not attached to the same substrate (e.g., manifold). In other cases, the valves (e.g., two-way valves, three-way valves) are attached to a manifold. In other words, the flow control circuit may be composed of manifold valves composed of individual valves. In some embodiments, the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is furthest from the outlet of the flow control circuit.The outlet of the flow control circuit may be defined as the position where the fluid exits the valve closest to one fluid connection with respect to the fluid path, or the position closest to this position. In other words, the inert flush fluid connection is provided at the position farthest from the outlet of the flow control circuit so as to maximize the flushing of other paths in order to minimize the dead volume and fluid carry-over between different flowing fluids.

[0028] In other cases, the flow control circuit has a multi-port selection valve. The multi-port selection valves described herein may have 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, such as 10 or more ports. The multi-port selection valves described herein may be configured to selectively enable access between ports. For example, the multi-port selection valve may be connected to a primary flush fluid line and one or more secondary fluid lines, but only one type of fluid can pass through at a time. In some embodiments, the internal volume of the multi-port selection valve is small. For example, the internal volume of the multi-port selection valve is 3 ~1,000 mm 3 、for example 20mm 3 ~100 mm 3 、for example 500 mm 3 ~1000mm 3 within the range of. Since the internal volume is small, it can be guaranteed that the entire multi-port selection valve is completely swept by the primary flush fluid and / or secondary fluid.

[0029] The fluid supply system of the present invention further includes one fluid connection configured to fluidly couple a flow control circuit, a working fluid line, and a fluid component of a device. The "one" fluid connection means that the fluid from the flow control circuit and the working fluid line 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 line can pass before being supplied to the fluid component of the device. In other words, one fluid connection is shared between the working fluid reservoir and the flow control circuit, and there is no additional non-cleanable tubing or path between the working fluid reservoir and the fluid connection. For this reason, the primary flush fluid and / or the secondary fluid can perform 100% flushing of the path between the working fluid reservoir and the device due to very low carryover. The one fluid connection may take any suitable form. In some cases, the one fluid connection is a quick connect fluid connector. In some such cases, the one fluid connection may be manually changed between the working fluid reservoir and the flow control circuit. Suitable connectors include, but are not limited to, snap-type (ball latch) connectors, bayonet connectors, threaded connectors, non-latch connectors, single shut-off connectors, double shut-off connectors, non-shut-off connectors, drive break connectors, roller lock connectors, pin lock connectors, ring lock connectors, and cam lock connectors. In other cases, the one fluid connection is a valve such as a three-way valve. In some embodiments, the internal volume of the three-way valve is small. For example, the internal volume of the three-way valve is in the range of 5 mm 3 to 100 mm 3 , for example 15 mm 3 to 30 mm 3 , for example 50 mm 3 to 80 mm 3 . Since the internal volume is small, it can be ensured that the entire three-way valve can be completely swept by the primary flush fluid and / or the secondary fluid. When the one fluid connection is a valve, the valve may be configured for automatic selection, i.e., configured such that the selection can be made between the working fluid reservoir and the flow control circuit without manual intervention.

[0030] In some cases, the fluid supply system includes a flow control circuit and a fluid supply pump operably connected to one fluid connection. The fluid supply pump may be configured to draw fluid from either the primary flush fluid reservoir or one or more secondary fluid reservoirs according to the settings 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 trapping a fixed volume of fluid and forcing (moving) the trapped fluid out of the device, and such a pump may operate in a series of operating cycles, trapping a fixed volume of fluid in each cycle and moving the fluid mechanically through the pump into the fluid system. Positive displacement pumps that may be used include rotary positive displacement pumps, such as peristaltic pumps, internal gear pumps, screw pumps, shuttle block pumps, flexible vane pumps, sliding vane pumps, circumferential piston pumps, flexible impeller pumps, helical twist roots pumps, or liquid-sealed 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, but are not limited thereto. In one embodiment, the positive displacement pump includes a pump selected from the group consisting of peristaltic pumps, gear pumps, and diaphragm pumps. In some cases, the positive displacement pump is a peristaltic pump.

[0031] FIG. 1 is a block diagram showing a fluid supply system according to an embodiment of the present invention. The fluid supply system 100 includes a working fluid reservoir 101, a primary flash fluid reservoir 102, and a set of secondary fluid reservoirs 103. Both the primary flash fluid reservoir 102 and the secondary fluid reservoirs 103 are fluidly connected to a flow control circuit 104 configured to selectively enable the primary flash fluid and the secondary fluid to pass through an outlet. The outlet of the flow control circuit 104 is fluidly coupled to a fluid supply pump 105 that draws fluid from one of the primary flash fluid reservoir 102 or the secondary fluid reservoirs 103 according to the setting of the flow control circuit 104. The fluid supply system 100 further includes one fluid connection portion 106, and the one fluid connection portion 106 may be fluidly connected to the flow control circuit 104 (via the fluid supply pump 105) or may be fluidly connected to the working fluid reservoir 101. The one fluid connection portion 106 enables either the movement of the working fluid from the working fluid reservoir 101 to the device 107 or the movement of the fluid from the flow control circuit 104 to the device 107, but does not enable both movements.

[0032] FIG. 2 is a diagram showing the fluid flow of a fluid supply system according to an embodiment of the present invention. The fluid supply system 200 includes a working fluid reservoir 201, a primary flash fluid reservoir 202, and a set of secondary fluid reservoirs 203. As the set of secondary fluid reservoirs 203, a first secondary fluid reservoir 203a, a second secondary fluid reservoir 203b, and an nth secondary fluid reservoir 203n are shown. The number of the second secondary fluid reservoirs in the set of secondary fluid reservoirs 203 may vary such that n is, for example, in the range of 3 to 10. The primary flash fluid line 208 fluidly couples the primary flash fluid reservoir 202 to the flow control circuit 204, while the secondary fluid lines 209a, 209b, and 209n fluidly 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 is understood that additional lines may be provided between the secondary fluid lines 209b and 209n according to the number of the set of secondary fluid reservoirs 203. The outlet of the flow control circuit 204 is fluidly coupled to a fluid supply pump 205 that draws fluid from either the primary flash fluid reservoir 202 or one of the set of secondary fluid reservoirs 203 according to the setting of the flow control circuit 204. The fluid supply system 200 further includes one fluid connection portion 206, and the one fluid connection portion 206 may be fluidly connected to the flow control circuit 204 (via the fluid supply pump 205) or may be fluidly connected to the working fluid reservoir 201. The one fluid connection portion 206 enables either the movement of the working fluid from the working fluid reservoir 201 to the device 207 or the movement of the fluid from the flow control circuit 204 to the device 207 at a given time, but does not enable both movements.

[0033] The system of the present invention may be configured to flush the working fluid through any suitable means to the fluid components (e.g., flow cell) of the device. In an embodiment of the system, a pressurized air source operably connected to the working fluid reservoir is provided. In such an embodiment, the pressurized air source may be used to apply a positive pressure to the working fluid reservoir to flush the working fluid contained in the working fluid reservoir through the fluid supply system, for example, to the fluid components of the device. The pressurized air source includes, for example, a compressor and a canister containing a pressurized gas. Any suitable compressor may be included. In some embodiments, the compressor is a positive displacement compressor, such as a rotary compressor (e.g., lobe compressor, screw compressor, liquid ring compressor, scroll compressor, vane compressor) or a reciprocating compressor (e.g., diaphragm compressor, double-acting compressor, single-acting compressor). In other cases, the compressor is a dynamic compressor such as a centrifugal compressor or an axial flow compressor. The system may further include a pressure regulator operably connected to the working fluid reservoir and the pressurized air source. Any suitable pressure regulator may be used, and it can include, for example, a regulator with a poppet valve, a diaphragm chamber regulator, a balanced poppet regulator, and a precision regulator. The pressure regulator of interest may be configured to adjust the flow rate of the flow stream by adjusting the pressure of the pressurized air applied to the working fluid reservoir.

[0034] FIG. 3 is a diagram showing the fluid flow of a fluid supply system 300 driven by pressurized air with respect to a liquid. The fluid supply system 300 includes a working fluid reservoir 301, a primary flash fluid reservoir 302, a set of secondary fluid reservoirs 303 (including secondary fluid reservoirs 303a, 303b, and 303n), a primary flash fluid line 308, secondary fluid lines 309a - 309n, a flow control circuit 304, a fluid supply pump 305, one fluid connection 306, a device 307, and a working fluid line 310. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 300 further 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 flush the working fluid contained in the working fluid reservoir through the working fluid line 310 and one fluid connection 306 to the device 307. The pressure regulator 311 is configured to control the amount of pressure applied and the flow rate of the working fluid.

[0035] In a further case, the system includes a working fluid supply pump configured to draw fluid from the working fluid reservoir and supply the working fluid to the fluid components of the device (e.g., a flow cell). The working fluid supply pump may be a positive displacement pump. Positive displacement pumps that may be used include rotary positive displacement pumps, such as peristaltic pumps, internal gear pumps, screw pumps, shuttle block pumps, flexible vane pumps, sliding vane pumps, circumferential piston pumps, flexible impeller pumps, helical twist roots pumps, or liquid-sealed 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, but are not limited thereto. In certain embodiments, the positive displacement pump includes a pump selected from the group consisting of peristaltic pumps, gear pumps, and diaphragm pumps. In some cases, the positive displacement pump is a peristaltic pump.

[0036] FIG. 4 is a diagram showing the fluid flow of a fluid supply system 400 including a working fluid supply pump according to an embodiment of the present invention. The fluid supply system 400 includes a working fluid reservoir 401, a primary flash fluid reservoir 402, a set of secondary fluid reservoirs 403 (including a secondary fluid reservoir 403a, a secondary fluid reservoir 403b, and a secondary fluid reservoir 403n), a primary flash fluid line 408, secondary fluid lines 409a to 409n, a flow control circuit 404, a fluid supply pump 405, one fluid connection 406, a device 407, and a working fluid line 410. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 400 further includes a working fluid supply pump 411 disposed between the device 407 and the one fluid connection 406. The working fluid supply pump 411 is configured to draw the working fluid from the working fluid reservoir 401 and supply it to the device 407.

[0037] In other cases, the fluid supply system includes a vacuum source operably connected to the fluid components of the device. In some cases, the vacuum source is a pump. The vacuum source may be disposed on the opposite side of the fluid component (e.g., flow cell) of the device with respect to the one fluid connection such that the vacuum generated by the vacuum source draws the working fluid from the working fluid reservoir through the one fluid connection and the fluid components of the device (e.g., flow cell). Any pump suitable for generating a vacuum capable of pushing the fluid from the working fluid reservoir may be used, including but not limited to the pumps described herein.

[0038] FIG. 5 is a diagram showing the fluid flow of a fluid supply system 500 including a vacuum source according to an embodiment of the present invention. The fluid supply system 500 includes a working fluid reservoir 501, a primary flash fluid reservoir 502, a set of secondary fluid reservoirs 503 (including secondary fluid reservoirs 503a, 503b, and 503n), a primary flash fluid line 508, secondary fluid lines 509a-509n, a flow control circuit 504, a fluid supply pump 505, one fluid connection 506, a device 507, and a working fluid line 510. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 500 further includes a vacuum source 511, and the vacuum source 511 is arranged on the opposite side of the device 507 with respect to one fluid connection such that the vacuum generated by the vacuum source 511 draws the working fluid from the working fluid reservoir 501 through one fluid connection and the device 507.

[0039] FIGS. 6A and 6B are diagrams showing the fluid flow of a fluid supply system 600 including one fluid connection constituted by a quick-connect fluid connector according to an embodiment of the present invention. The fluid supply system 600 includes a working fluid reservoir 601, a primary flash fluid reservoir 602, a set of secondary fluid reservoirs 603 (including secondary fluid reservoirs 603a, 603b, and 603n), a primary flash fluid line 608, secondary fluid lines 609a-609n, a flow control circuit 604, a fluid supply pump 605, a device 607, and a working fluid line 610. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 600 further includes one fluid connection constituted by a quick-connect fluid connector 606 that may be fluidly coupled to either the working fluid reservoir 601 or the flow control circuit 604 at a given time. FIG. 6A shows the quick-connect fluid connector 606 fluidly coupled to the working fluid reservoir 601, while FIG. 6B shows the quick-connect fluid connector 606 fluidly coupled to the flow control circuit 604.

[0040] FIG. 7 is a diagram showing the fluid flow of a fluid supply system 700 including one fluid connection portion constituted by a three-way valve according to an embodiment of the present invention. The fluid supply system 700 includes a working fluid reservoir 701, a primary flush fluid reservoir 702, a set of secondary fluid reservoirs 703 (including a secondary fluid reservoir 703a, a secondary fluid reservoir 703b, and a secondary fluid reservoir 703n), a primary flush fluid line 708, secondary fluid lines 709a to 709n, a flow control circuit 704, a fluid supply pump 705, a device 707, and a working fluid line 710. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 700 further includes one fluid connection portion 706 constituted by a three-way valve 706. The three-way valve 706 has two inlets, one of which is fluidly connected to the working fluid reservoir 701 and the other of which is fluidly connected to the flow control circuit 704, and an outlet fluidly connected to the device 707. Depending on the setting of the three-way valve 706, fluid from either the working fluid reservoir 701 or the flow control circuit 704 may be supplied to the device 707. In some cases, the three-way valve 706 may be automatically controlled, for example, by a processor.

[0041] FIG. 8 is a diagram showing the fluid flow of a fluid supply system 800 including a flow control circuit composed of a series of stand-alone two-way valves according to an embodiment of the present invention. The fluid supply system 800 includes a working fluid reservoir 801, a primary flush fluid reservoir 802, a set of secondary fluid reservoirs 803 (including secondary fluid reservoirs 803a, 803b, and 803n), a primary flush fluid line 808, secondary fluid lines 809a-809n, a fluid supply pump 805, one fluid connection 806, a device 807, and a working fluid line 810. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 800 further includes a flow control circuit composed of a series of stand-alone two-way valves 804a-804d. Each of the two-way valves 804a-804d may be individually operable such that fluid can be supplied from the respective reservoir when the two-way valve is open, while this supply is blocked when the two-way valve is closed. The opening and closing of the two-way valve may in some cases be automatically controlled, for example, by a processor. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines may be provided in the fluid supply system 800 (as described above). In this case, an additional two-way valve may be provided for each additional secondary fluid reservoir and secondary fluid line. As shown in FIG. 8, the primary flush fluid line 808 is fluidly coupled to the two-way valve 804a, which is the farthest two-way valve in a row from the outlet of the flow control circuit (where the two-way valves 804a-804d are fluidly connected to the fluid supply pump 805). For this reason, it is ensured that the fluid lines of the flow control circuit are completely flushed by the primary flush fluid.

[0042] FIG. 9 is a diagram showing the fluid flow of a fluid supply system 900 including a flow control circuit composed of a series of two-way valves commonly attached according to an embodiment of the present invention. The fluid supply system 900 includes a working fluid reservoir 901, a primary flush fluid reservoir 902, a set of secondary fluid reservoirs 903 (including secondary fluid reservoirs 903a, 903b, and 903n), a primary flush fluid line 908, secondary fluid lines 909a to 909n, a fluid supply pump 905, one fluid connection 906, a device 907, and a working fluid line 910. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 900 further includes a flow control circuit composed of a series of two-way valves 904a to 904d attached to a manifold 904. Each of the two-way valves 904a to 904d may be individually operable such that fluid can be supplied from each reservoir when the two-way valve is open, while this supply is blocked when the two-way valve is closed. The opening and closing of the two-way valve may be automatically controlled by, for example, a processor in some cases. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines may be provided in the fluid supply system 900 (as described above). In this case, an additional two-way valve may be provided in the manifold 904 for each additional secondary fluid reservoir and secondary fluid line. As shown in FIG. 9, the primary flush fluid line 908 is fluidly coupled to the two-way valve 904a in the manifold 904 that is farthest from the outlet of the flow control circuit (where the two-way valves 904a to 904d are fluidly connected to the fluid supply pump 905). For this reason, it is guaranteed that the fluid lines of the flow control circuit are completely flushed by the primary flush fluid.

[0043] FIG. 10 is a diagram showing the fluid flow of a fluid supply system 1000 including a flow control circuit composed of a row of stand-alone three-way valves according to an embodiment of the present invention. The fluid supply system 1000 includes a working fluid reservoir 1001, a primary flush fluid reservoir 1002, a set of secondary fluid reservoirs 1003 (including a secondary fluid reservoir 1003a, a secondary fluid reservoir 1003b, and a secondary fluid reservoir 1003n), a primary flush fluid line 1008, secondary fluid lines 1009a to 1009n, fluid supply pumps 1005, one fluid connection 1006, a device 1007, and a working fluid line 1010. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 1000 further includes a flow control circuit composed of stand-alone three-way valves 1004a, 1004b, and 1004c. The three-way valve 1004a has one inlet fluidly connected to the primary flush fluid reservoir 1002 via the primary flush fluid line 1008 and another inlet fluidly connected to the secondary fluid reservoir 1003a. The three-way valve 1004b has one inlet fluidly connected to the outlet of the three-way valve 1004a and another inlet fluidly connected to the secondary fluid reservoir 1003b via the secondary fluid line 1009a. The three-way valve 1004c has one inlet fluidly connected to the outlet of the three-way valve 1004b and another inlet fluidly connected to the secondary fluid reservoir 1003n via the secondary fluid line 1009n. It should be understood that one or more additional secondary fluid reservoirs and secondary fluid lines may be provided in the fluid supply system 1000 (as described above, for example). In this case, an additional three-way valve may be provided for each additional secondary fluid reservoir and secondary fluid line. Each of the three-way valves 1004a to 1004c may be individually operable such that fluid can be supplied from each reservoir when the three-way valve is open, while this supply is blocked when the three-way valve is closed.As shown in FIG. 10, the primary flash fluid line 1008 is fluidly coupled to the three-way valve 1004a that is the farthest from the outlet of the flow control circuit (where the three-way valves 1004a to 1004c are fluidly connected to the fluid supply pump 1005). For this reason, it is guaranteed that the fluid lines of the flow control circuit are completely flushed by the primary flash fluid.

[0044] FIG. 11 is a diagram showing the fluid flow of a fluid supply system 1100 including a flow control circuit composed of a series of three-way valves attached to a manifold according to an embodiment of the present invention. The fluid supply system 1100 includes a working fluid reservoir 1101, a primary flush fluid reservoir 1102, a set of secondary fluid reservoirs 1103 (including secondary fluid reservoir 1103a, secondary fluid reservoir 1103b, and secondary fluid reservoir 1103n), a primary flush fluid line 1108, secondary fluid lines 1109a to 1109n, fluid supply pumps 1105, one fluid connection 1106, a device 1107, and a working fluid line 1110. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 1100 further includes a flow control circuit composed of three-way valves 1104a, 1104b, and 1104c attached to the manifold 1104. The three-way valve 1104a has one inlet fluidly connected to the primary flush fluid reservoir 1102 via the primary flush fluid line 1108 and another inlet fluidly connected to the secondary fluid reservoir 1103a via the secondary fluid line 1109a. The three-way valve 1104b has one inlet fluidly connected to the outlet of the three-way valve 1104a and another inlet fluidly connected to the secondary fluid reservoir 1103b via the secondary fluid line 1109b. The three-way valve 1104c has one inlet fluidly connected to the outlet of the three-way valve 1104b and another inlet fluidly connected 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 may be provided in the fluid supply system 1100 (as described above, for example). In this case, an additional three-way valve may be provided in the manifold for each additional secondary fluid reservoir and secondary fluid line. Each of the three-way valves 1104a to 1104c may be individually operable such that fluid can be supplied from the respective reservoirs when the three-way valve is open, while this supply is blocked when the three-way valve is closed.As shown in FIG. 11, the primary flush fluid line 1108 is fluidly coupled to the three-way valve 1104a that is farthest from the outlet of the flow control circuit (where the three-way valves 1104a to 1104c are fluidly connected to the fluid supply pump 1105). For this reason, it is ensured that the fluid lines of the flow control circuit are completely flushed by the primary flush fluid.

[0045] FIG. 12 is a diagram showing the fluid flow of a fluid supply system 1200 including a flow control circuit composed of a multi-port selection valve according to an embodiment of the present invention. The fluid supply system 1200 includes a working fluid reservoir 1201, a primary flush fluid reservoir 1202, a set of secondary fluid reservoirs 1203 (including secondary fluid reservoirs 1203a, 1203b, and 1203n), a primary flush fluid line 1208, secondary fluid lines 1209a to 1209n, fluid supply pumps 1205, one fluid connection 1206, a device 1207, and a working fluid line 1210. The above elements are arranged as described above with respect to FIG. 2. The fluid supply system 1200 further includes a multi-port selection valve 1204. The multi-port selection valve 1204 is fluidly connected to the primary flush fluid reservoir 1202 via the primary flush fluid line 1208, fluidly connected to the secondary fluid reservoir 1203a via the secondary fluid line 1209a, fluidly connected to the secondary fluid reservoir 1203b via the secondary fluid line 1209b, fluidly connected to the secondary fluid reservoir 1203n via the secondary fluid line 1209n, and fluidly connected to the fluid supply pump 1205. The multi-port selection valve 1204 is configured to selectively send 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 may be provided in the fluid supply system 1200 (as described above, for example). In this case, these reservoirs and secondary fluid lines may be similarly fluidly connected to the multi-port selection valve 1204.

[0046] Fluid management system Aspects of the present invention further include a fluid management system comprising the fluid supply system of the present invention. The term "fluid management system" refers to the entire fluid components associated with an instrument, such as a flow cytometer. The fluid management system of the present disclosure may be configured to perform one or more functions, including supplying a working fluid (e.g., sheath fluid) and a sample fluid to a fluid component (e.g., flow cell) at a controlled ratio, supplying a primary flush fluid to the fluid component, supplying one or more secondary fluids (e.g., for washing or as a reagent) to the fluid component, hydrodynamically focusing the sample fluid using the working fluid for sample interrogation through a cuvette, collecting the analyzed sample as waste, collecting waste by sorting samples, and managing and storing all system waste, but is not limited thereto. The fluid management system may include a flow cell having an input and an output, a cuvette having an input coupled to the output of the flow cell and further having an output, a sample input line for fluidly coupling a sample fluid source to the input of the flow cell, and any of the fluid supply systems described herein. Exemplary fluid management systems that may be adapted to include the subject fluid supply system include the fluid management system described in U.S. Patent Application Publication No. 2022 / 0341838, the disclosure of which is incorporated herein by reference.

[0047] FIG. 13 is a flowchart showing the main functions of a fluid management system configured to analyze a sample. The main functions are: (1301) supplying the working fluid (e.g., sheath) and the sample fluid to the flow cell at a controlled ratio; (1302) supplying any number of primary flush fluids and secondary fluids (e.g., for cleaning); (1303) hydrodynamically focusing the sample using the working fluid for sample interrogation through the cuvette; (1304) collecting the analyzed sample as waste; and (1305) managing and storing all system waste (waste from particle analysis 1304 and secondary system waste 1306). The dotted lines indicate that (a) the working fluid and the sample fluid may be supplied to the flow cell, or (b) any number of secondary system fluids may be supplied to the flow cell.

[0048] In certain embodiments, the fluid management system is alternatively configured to (a) analyze a sample (e.g., particles in the sample fluid), or (b) analyze and sort a sample (e.g., particles in the sample fluid). In such embodiments, the fluid management system may include all of the components necessary to (a) analyze the sample or (b) analyze and sort the sample, but only one of (a) and (b) can be performed at a time. In some embodiments, the fluid management system is configured to be able to (a) analyze a sample and (b) analyze and sort a sample, but in some cases (a) the analysis of the sample is performed, and in other cases (b) the analysis and sorting of the sample is performed.

[0049] Aspects of fluid management by a fluid management system may be based on the principles of a fluid circuit. For example, the basic principle of the supply of the working fluid may be based on the principles of a fluid circuit which assumes that the pressure drop in a closed fluid path is equal to the product of the liquid flow rate and the fluid resistance. The paths of the working fluid (e.g., sheath) and the sample can be modeled as two parallel resistors and combined to proceed through a flow cell, a cuvette, and a waste path which are the third fluid resistors. Similarly, waste management using one waste pump according to certain embodiments is based on the principles of a fluid circuit. All individual waste suction sources can be modeled as fluid reservoirs at atmospheric pressure, and all connections (e.g., fluid lines) from these waste sources to the waste management system can be modeled as fluid resistors. Considering the supply vacuum of the waste pump, as described in more detail below, the fluid resistance of each connection can be controlled to balance the required waste suction force of each waste source.

[0050] The waste fluid of the flow-type particle analyzer of the present disclosure may be managed by a fluid transfer device according to certain embodiments. In some cases, the waste fluid of the fluid management system, such as the mixed waste fluid, is managed by one fluid transfer device, such as one waste pump. The mixed waste fluid may be managed by one fluid transfer device, such as one waste pump, by taking an appropriate balance of fluid resistance from all of the various waste sources of the system, such as analyzer waste, sorted waste, and secondary system waste. The waste sources may include, for example, an analyzer waste fluid recovery subsystem, a sorter waste fluid recovery subsystem, and one or more system waste fluid sources. The connections (e.g., fluid lines) from the waste sources may function as a plurality of fluid resistors in parallel with one pressure source, such as a waste vacuum pump, which is a fluid transfer device. As long as the fluid transfer device, such as the waste vacuum pump, has sufficient capacity, the resistance can be adjusted so that each waste source has an appropriate vacuum. For example, the resistance of the flow path (e.g., fluid line) fluidly coupling the waste sources of the system to the fluid transfer device may have a balanced known resistance so that each waste source can have an appropriate vacuum to draw out the waste fluid. In some cases, the resistance of the first fluid line coupled to the first waste source among the plurality of fluid lines coupling the plurality of waste sources to the fluid transfer device may be adjusted or set to provide an appropriate vacuum to the first waste source. The resistance of the second fluid line coupled to the second waste source among the plurality of fluid lines coupling the plurality of waste sources to the fluid transfer device may be adjusted or set to provide an appropriate vacuum to the second waste source. The resistance of the additional fluid lines in the system may further have individual resistances that are adjusted or set to provide an appropriate vacuum for each of the respective waste sources. The resistance of the fluid lines from each of the waste sources may be the same or different. Each fluid line fluidly coupling the waste sources to the fluid transfer device may have a specific known resistance so that each of the waste sources has an appropriate vacuum.For example, the resistance of fluid lines coupling an analyzer waste fluid recovery subsystem, a sorter waste fluid recovery subsystem, and one or more of one or more system waste fluid sources to a fluid moving device may be balanced to enable appropriate vacuum for each waste source and control of suction of waste fluid by one fluid moving device. In some cases, the resistance of multiple fluid lines coupling multiple waste sources to a fluid moving device may be individually adjusted to provide fluid from each waste source at a desired flow rate. For example, the resistance of a first fluid line coupled to a first waste source among multiple fluid lines coupling multiple waste sources to a fluid moving device may be adjusted or set to provide waste fluid from the first waste source at a desired first flow rate. The resistance of a second fluid line coupled to a second waste source among multiple fluid lines coupling multiple waste sources to a fluid moving device may be adjusted or set to provide waste fluid from the second waste source at a desired second flow rate. The resistance of additional fluid lines within the system may further have individual resistances that are adjusted or set to provide waste fluid from each of the respective waste sources at a desired flow rate. The resistance of fluid lines from each of the waste sources may be the same or different. Each fluid line fluidly coupling a waste source to a fluid moving device may have a specific known resistance based on the desired flow rate of fluid from each of the waste sources. In some cases, the resistance of a flow path (e.g., a fluid line) within a fluid management system is constant. In some cases, the resistance of a flow path (e.g., a fluid line) within a fluid management system is variable. In some cases, the resistance of one or more flow paths within a fluid management system may be independently variable, e.g., the resistance of each flow path may vary individually. In some cases, the fluid management system includes one or more variable resistors that may adjust the resistance of a flow path, e.g., by adjusting the dimensions of the flow path (e.g., the diameter or length of a fluid line). In some cases, the variable resistor has a valve.

[0051] The fluid management system of the present disclosure may further include one or more waste fluid recovery subsystems. In some cases, the fluid management system includes an analyzer waste fluid recovery subsystem configured to fluidly couple the output of the cuvette to a waste management subsystem, a sorter waste fluid recovery subsystem configured to fluidly couple the output of a sorter block coupled to the output of the cuvette to the waste management subsystem, or combinations thereof (e.g., in a hybrid system where a flow-through particle analyzer may (a) analyze a sample or (b) analyze and sort a sample).

[0052] The waste management subsystem may be configured to receive and mix waste fluids from (a) one or more system waste fluid sources and (b) an analyzer waste fluid recovery subsystem, a sorter waste fluid recovery subsystem, or a combination thereof to produce a mixed waste fluid. The fluid from the system waste fluid source may include waste fluid generated and / or recovered by cleaning any part of the flow-through particle analyzer. The fluid from the analyzer waste fluid recovery subsystem may include, for example, analyzed waste fluid from the output of a cuvette that may include analyzed sample fluid and actuation fluid that has flowed through the cuvette and been optically investigated. The fluid from the sorter waste fluid recovery subsystem may include waste fluid received from the output of the sorting block, for example, the output of the sorting nozzle. The waste fluid may include analyzed actuation fluid and sample fluid that has not been conveyed, for example, in droplets, to the sample collection container by one or more deflection plates of the sorting block (e.g., including non-target particles). In certain embodiments, the waste management subsystem is configured to receive and mix waste fluids from (a) one or more system waste fluid sources and (b) an analyzer waste fluid recovery subsystem to produce a mixed waste fluid. In certain embodiments, the waste management subsystem is configured to receive and mix waste fluids from (a) one or more system waste fluid sources and (b) a sorter waste fluid recovery subsystem to produce a mixed waste fluid. In certain embodiments, the waste management subsystem is configured to receive and mix waste fluids from (a) one or more system waste fluid sources, (b) an analyzer waste fluid recovery subsystem, and a sorter waste fluid recovery subsystem. In certain embodiments, the waste management subsystem is configured to receive and mix waste fluids from (a) one or more system waste fluid sources and (b) an analyzer waste fluid recovery subsystem or a sorter waste fluid recovery subsystem.

[0053] The waste management subsystem may be fluidly coupled to (a) one or more system waste fluid sources and (b) an analyzer waste fluid recovery subsystem, a sorter waste fluid recovery subsystem, or a combination thereof. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste fluid sources and (b) an analyzer waste fluid recovery subsystem. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste fluid sources and (b) a sorter waste fluid recovery subsystem. In certain embodiments, the waste management subsystem is fluidly coupled to (a) one or more system waste fluid sources and (b) both an analyzer waste fluid recovery subsystem and a sorter waste fluid recovery subsystem.

[0054] The waste management subsystem may have an output for the mixed waste fluid, such as one output. Optionally, the output of the waste management subsystem is fluidly coupled to a waste fluid container. A fluid line, such as tubing or a pipe, may fluidly couple the output of the waste management subsystem to the waste container. Optionally, the output is fluidly coupled to the waste container with one fluid line. The mixed waste fluid may flow, for example, from the output into the waste fluid container for storage. The waste container may be fluidly coupled, for example via a fluid line (e.g., one fluid line), to a fluid transfer device configured to carry the mixed waste fluid from the output of the waste management subsystem to the waste fluid container. Optionally, the waste fluid container is detachable from a fluid line that couples the waste fluid container to the fluid management system, for example to be able to empty and clean the waste fluid container.

[0055] If desired, the fluid management system may include a fluid transfer device configured to transfer, e.g., mechanically, the mixed waste fluid to the waste fluid container. In some cases, the mixed waste fluid is managed by one fluid transfer device, e.g., one pump. In these embodiments, since only one fluid transfer device is provided, the fluid management system does not include any other fluid transfer device for controlling or regulating the flow of the mixed waste fluid to the waste container. In some embodiments, one fluid line may couple the output of the waste management subsystem to the waste container. In some embodiments, one fluid line fluidly couples the output of the waste management subsystem to one fluid transfer device, and the one fluid transfer device is then fluidly coupled to the waste fluid container by one fluid line. In some cases, the mixed waste fluid is transferred by one fluid transfer device, e.g., one pump, from the output of the waste management subsystem to the waste fluid container. In some cases, the fluid transfer device is a vacuum source that draws the mixed waste fluid from the output of the fluid management subsystem to the waste fluid container. In some cases, the fluid transfer device transports waste fluid, e.g., mixes it, from each of the subsystem or waste source (e.g., analyzer waste fluid recovery subsystem, sorter waste fluid recovery subsystem, one or more system waste fluid sources) to the waste management subsystem and then transports the mixed waste fluid to the waste container. In some cases, the waste fluid from each of the subsystems or waste sources fluidly coupled to the waste management subsystem is managed by one fluid transfer device. In some cases, the fluid transfer device is a vacuum source that draws the waste fluid from each of the subsystem or waste source to the waste management subsystem. The fluid transfer device may be fluidly coupled to the output of the waste management subsystem and the waste fluid container. The fluid transfer device may be any fluid transfer device as described herein. In some cases, the fluid transfer device is a vacuum generating device. In some cases, the fluid transfer device has a pump, e.g., a vacuum pump. In some cases, the fluid transfer device is one pump. In some cases, the pump has a positive displacement vacuum pump. In some cases, the positive displacement vacuum pump has 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.

[0056] In some embodiments, the flow-through particle analyzer has a waste fluid container. The waste container may be any suitable reservoir or container (e.g., having a rigid or flexible wall) for storing the waste fluid. In some cases, the output of the waste management subsystem is fluidly coupled to the waste fluid container. In some cases, the waste container is fluidly coupled to a fluid transfer device configured to carry the mixed waste fluid from the output of the waste management subsystem to the waste fluid container. A fluid line, such as tubing or piping, may fluidly couple the output of the waste management subsystem to the waste container. In some cases, the output is fluidly coupled to the waste container with one fluid line. The mixed waste fluid may flow, for example, from the output into the waste fluid container for storage. The waste container may be fluidly coupled, for example, via a fluid line (e.g., one fluid line) to a fluid transfer device configured to carry the mixed waste fluid from the output of the waste management subsystem to the waste fluid container. In some cases, the waste fluid container is detachable from the fluid line that couples the waste fluid container to the fluid management system, for example, so that the waste fluid container can be emptied and cleaned. The waste fluid container may be configured to have a suitable volume, for example, to hold and store all of the system fluid. The volume of the waste fluid container may be in the range of 1 L to 100 L, for example, the volume of the container may be in the range of 1 L to 90 L, 1 L to 80 L, 1 L to 70 L, 1 L to 60 L, 1 L to 50 L, 1 L to 40 L, 1 L to 30 L, 1 L to 20 L, or 1 L to 10 L.

[0057] The fluid management system may comprise one or more system waste fluid sources that are fluidly coupled to the waste management subsystem. The one or more system waste fluid sources may generate and / or recover waste fluid by cleaning any area or location of the flow-type particle analyzer. The one or more system waste fluid sources may be configured to carry or convey the system waste fluid to the waste management subsystem. In some cases, the waste fluid from the one or more system waste fluid sources is drawn into the waste management subsystem by a vacuum generated by a fluid moving device. The one or more waste fluid sources may be fluidly coupled to the waste management subsystem with a known fluid resistance. In some cases, one or more fluid lines coupling the one or more system waste fluid sources to the waste management subsystem, for example, each of the one or more fluid lines is provided with a resistor. The one or more resistors may provide a known fluid resistance to the one or more fluid lines coupling the one or more system waste fluid sources to the waste management subsystem. Suitable resistors include, but are not limited to, a portion of tubing having a known length and inner diameter, such as an orifice.

[0058] The fluid management system of the present disclosure may further comprise an operating fluid pressure feedback control subsystem configured to control or adjust the operating fluid pressure in a system, such as an operating fluid reservoir. The operating fluid pressure may be controlled by an adjusted air pressure in the operating fluid reservoir. The pressurized operating fluid may flow from an operating fluid source through a fluid supply system to a flow cell. The operating fluid pressure may be controlled based on various parameters, such as the measured liquid level height in the operating fluid reservoir and the air pressure in the operating fluid reservoir. Additional parameters may include, but are not limited to, the flow rate of the operating fluid.

[0059] The actuated fluid pressure feedback control subsystem may have a control unit configured to maintain a controlled variable, such as fluid pressure, at a desired setpoint. The control unit may be any suitable control system. In some cases, the control unit has a closed-loop control system, such as a feedback control unit. The system may have any suitable sensor for detecting one or more parameters for the feedback signal. In some cases, the control unit is configured to receive a signal representing the measured actuated fluid reservoir air pressure. In some cases, the control unit is configured to receive a signal representing the measured liquid level height in the actuated fluid reservoir. In some cases, the control unit is configured to receive a signal representing the measured actuated fluid reservoir air pressure and a signal representing the measured liquid level height in the actuated fluid reservoir. In some cases, the control unit is configured to determine whether the actuated fluid pressure has deviated from the setpoint based on the received signal representing the measured actuated fluid reservoir air pressure and / or the received signal representing the measured liquid level height in the actuated fluid reservoir. In some cases, the control unit transmits an actuated fluid pressure control signal to the actuated fluid reservoir air pressure regulator to adjust, for example, the actuated fluid reservoir air pressure if the actuated fluid pressure has deviated from the setpoint (thereby regulating the actuated fluid pressure and the actuated fluid flow rate). By the actuated fluid pressure control signal, the actuated fluid reservoir air pressure regulator may adjust the air pressure in the actuated fluid reservoir so that the actuated fluid pressure matches the desired setpoint. In some cases, the control unit is configured to transmit the actuated fluid pressure control signal to the control unit of the sample fluid pressure feedback control system. Any suitable feedback control mechanism may be used. The noted feedback control unit may use control parameters, such as a proportional gain, an integral term, and / or a derivative term. The control parameters may be applied to an error signal (e.g., the difference between the setpoint and the feedback signal) to calculate an input to be provided to the controlled system or process. In one embodiment, the control unit is a proportional integral derivative (PID) control unit.

[0060] In some cases, the working fluid pressure feedback control subsystem has a working fluid reservoir air pressure regulator configured to control the air pressure in the working fluid reservoir. The working fluid reservoir air pressure regulator may be configured to receive a working fluid pressure control signal from the control unit and adjust the air pressure in the working fluid reservoir (thereby adjusting the fluid pressure). The working fluid reservoir air pressure regulator may adjust the air pressure in the working fluid reservoir so that the working fluid pressure matches a desired set value. The working fluid reservoir air pressure regulator may be gas-coupled to a pressurized air source. In some cases, the working fluid reservoir air pressure regulator is gas-coupled to the working fluid reservoir. In some cases, the inlet of the working fluid reservoir air pressure regulator is gas-coupled to a pressurized air source. In some cases, the outlet of the working fluid reservoir air pressure regulator is gas-coupled to the working fluid reservoir. Any suitable air pressure regulator may be used. The pressure regulator of interest may include, for example, a device that controls (e.g., adjusts) a supply (or inlet) pressure to a desired outlet pressure and operates to maintain this outlet pressure despite fluctuations in the inlet pressure. In some cases, the working fluid reservoir air pressure regulator is an electro-pneumatic pressure regulator.

[0061] In some cases, the actuating fluid pressure feedback control subsystem has a liquid level measuring device configured to measure the liquid level height in the actuating fluid reservoir. The liquid level measuring device may be operably coupled to the actuating fluid reservoir. The output signal of the liquid level measuring device (e.g., representing the measured liquid level) may be transmitted to the control unit of the actuating fluid pressure feedback control subsystem according to the embodiments described herein. In some cases, the liquid level measuring device is a point level measuring sensor configured to mark one individual liquid level height. In some cases, the liquid level measuring device is a continuous level sensor configured to measure the liquid level within a certain range. Suitable liquid level measuring devices include, but are not limited to, glass liquid level gauges, float switches, ultrasonic sensors, capacitance level sensors, hydrostatic devices (e.g., bubblers, displacer, differential pressure transmitters), strain gauges, magnetic level gauges, magnetostrictive level transmitters, laser level transmitters, radar level sensors, inductive radar sensors, tuning forks, etc.

[0062] The fluid management system of the present disclosure may further include a sample fluid pressure feedback control subsystem configured to control or adjust the sample fluid pressure within the system, for example, within the sample fluid source. The sample fluid pressure may be controlled by the regulated air pressure within the sample fluid source. The pressurized sample fluid may be supplied directly from the sample fluid source to the flow cell. The fluid pressure within the sample fluid source may be controlled based on various parameters including, for example, the measured air pressure within the sample fluid source and the actuating fluid pressure control signal. Additional parameters may include, but are not limited to, the sample liquid level height and the flow rate of the sample fluid.

[0063] The sample fluid pressure feedback control subsystem may have a control unit configured to maintain a controlled variable, such as fluid pressure, at a desired setpoint. The control unit may be any suitable control system. In some cases, the control unit has a closed-loop control system, such as a feedback control unit. The system may have any suitable sensor for detecting one or more parameters for the feedback signal. The sample fluid pressure setpoint may be the sheath fluid pressure + the desired flow rate. In such a case, the sample fluid pressure will always be higher than the working fluid (e.g., sheath) pressure for a positive flow through the system. Thus, it is possible to dynamically adjust the working fluid without changing the effective pressure drop of the sample fluid. The systems described herein may control the sample pressure as a differential pressure above the working fluid (e.g., sheath) pressure. Thus, it is possible to vary the working fluid pressure while keeping the sample flow rate constant (P_sample (sample fluid pressure) = P_sheath (sheath fluid pressure) + P_diff (differential pressure)). In one embodiment, the control unit is configured to receive a signal representing the measured sample fluid source air pressure. In one embodiment, the control unit is configured to receive, for example, a working fluid pressure control signal from the control unit of the working fluid pressure feedback control subsystem. In one embodiment, the control unit is configured to receive a signal representing the measured sample fluid source air pressure and a working fluid pressure control signal. In some cases, the control unit is configured to determine whether the sample fluid pressure has deviated from the setpoint based on the received signal representing the measured sample fluid source air pressure and / or the working fluid pressure control signal. The control unit may transmit a sample fluid pressure control signal to the sample fluid source air pressure regulator to adjust, for example, the sample fluid source air pressure if the sample fluid pressure has deviated from the setpoint (thereby adjusting the sample fluid pressure and the sample fluid flow rate). In response to the sample fluid pressure control signal, the sample fluid source air pressure regulator may adjust the air pressure in the sample fluid source so that the sample fluid pressure matches the desired setpoint. Any suitable feedback control mechanism may be used. The noted feedback control unit may use control parameters such as a proportional gain, an integral term, and / or a derivative term.The control parameter may be applied to an error signal (e.g., the difference between a set value and a feedback signal) to calculate an input applied to a controlled system or process. In certain embodiments, the control unit is a PID control unit.

[0064] In some cases, the sample fluid pressure feedback control subsystem has a sample fluid source air pressure regulator configured to control the air pressure within the sample fluid source. The sample fluid source air pressure regulator may be configured to receive a sample fluid pressure control signal from the control unit and adjust the air pressure within the sample fluid source (thereby adjusting the fluid pressure). The sample fluid source air pressure regulator may adjust the air pressure within the sample fluid source such that the sample fluid pressure matches a desired set value. The sample fluid source air pressure regulator may be pneumatically coupled to a pressurized air source. In some cases, the sample fluid source air pressure regulator is pneumatically coupled to the sample fluid source. In some cases, the inlet of the sample fluid source air pressure regulator is pneumatically coupled to a pressurized air source. In some cases, the outlet of the sample fluid source air pressure regulator is pneumatically coupled to the sample fluid source. Any suitable air pressure regulator may be used. The pressure regulator of interest may include, for example, a device that controls (e.g., adjusts) a supply (or inlet) pressure to a desired outlet pressure and operates to maintain this outlet pressure despite fluctuations in the inlet pressure. In some cases, the sample fluid source air pressure regulator is an electro-pneumatic pressure regulator.

[0065] In certain embodiments, the flow-through particle analyzer has a sample fluid source. The sample fluid source may be any suitable reservoir or container (e.g., having a rigid or flexible wall) for holding 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 of the sample fluid container may be in the range of 1 mL to 100 mL, and for example, the volume of the container may be in the range of 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.

[0066] The sample fluid source may contain the sample fluid, and the pressurized sample fluid may flow from the sample fluid source to the flow cell. Pressure may be applied to the sample fluid using a pressurized air source and a sample fluid source air pressure regulator gas-coupled to the sample fluid source. The air pressure in the sample fluid source may be adjusted by a sample fluid pressure feedback control subsystem to control the pressure and flow rate of the sample fluid.

[0067] FIG. 14 is a flowchart showing sub-functions within the main functions of a fluid management system configured to analyze a sample according to an embodiment. The configuration is a configuration that manages the secondary system fluid and all waste fluids based on pumps. A fluid supply pump 1406 manages the primary flush system fluid and the secondary system fluid 1403. A waste pump 1412 manages the waste fluid from the discharge waste accumulation section 1411 and the system waste 1414.

[0068] In the fluid configuration, the sheath and the sample are sent based on positive pressure, and a positive pressure source is used to control the flow rate ratio of the sheath / sample liquid. The working fluid (sheath) is controlled by the regulated air pressure 1402, and the sample fluid pressure is controlled by the regulated air pressure 1404. The air pressure feedback control 1401 is performed in a closed loop via inputs from the working fluid (sheath) chamber 1405 and the sample chamber 1407. Examples of feedback parameters include measurements such as the air pressure in the chamber and the liquid level height for pressure head correction. Further, the working fluid (sheath) pressure control signal is used as feedback for sample pressure control. The sample pressure set value is always the sheath pressure + the desired flow rate. This is because the sample pressure is always higher than the working fluid (sheath) pressure for the positive flow through the system. Therefore, it is possible to dynamically adjust the working fluid without changing the effective pressure drop of the sample fluid.

[0069] The pressurized working fluid (sheath) travels through the fluid supply system 1409 to the flow cell 1408, and the pressurized sample is supplied directly to the flow cell 1408. Fluid 1403 in any number of fluid supply reservoirs (e.g., for cleaning fluid) can be further sent through the fluid supply pump 1406 to the fluid supply system 1409 in place of the working fluid (sheath) 1405. The dotted lines indicate that (a) the working fluid may be supplied to the flow cell, or (b) any number of secondary system fluids may be supplied to the flow cell. This enables 100% flushing of the fluid system. The sample is investigated as the working fluid and the sample fluid pass through the flow cell and cuvette 1408.

[0070] After the working fluid and the sample fluid (or secondary fluid during cleaning) have flowed through the flow cell, both fluids can be considered "analyzed waste fluid" 1410 that is recovered at atmospheric pressure in the discharge waste accumulation section 1411. All waste from the system (analyzed waste 1410 and any secondary waste sources 1414 such as from cleaning operations) are mixed and drawn out by one vacuum waste source such as the waste pump 1412. The mixed waste is sent to the waste storage section 1413.

[0071] FIG. 15 is a schematic diagram showing a fluid management system according to an embodiment. A pressurized air source 1501 is used to supply pneumatic pressure to an electro-pneumatic pressure regulator. The pressure regulator 1502 controls the air pressure in the working fluid reservoir 1504, and the sample fluid regulator 1503 controls the pressure in the sample chamber 1505. A feedback control mechanism 1506, such as a PID control unit, is used to transmit a control signal 1508 to the pressure regulator 1502. A similar feedback control mechanism 1507 is used to transmit a control signal 1509 to the sample fluid regulator 1503. The feedback of the working fluid pressure is obtained from the measured air pressure 1510 in the working fluid reservoir and the liquid level height 1513 in the working fluid reservoir measured using the liquid level measuring device 1512. The feedback of the sample fluid pressure is obtained from the measured air pressure 1511 in the sample chamber and the working fluid control signal 1508. The set value of the sample fluid is always controlled as a delta higher than the working fluid pressure so that the pressure drop in the sample line remains constant even when the working fluid pressure changes. The working fluid reservoir 1504 has a quick disconnect connection portion 1515 that connects to pipes 1516 and fitting connectors 1514. Any arbitrary amount of secondary fluid is stored in a set of secondary fluid reservoirs 1553. In addition, the primary flush fluid reservoir 1552 contains the primary flush fluid. The flow control circuit 1554 selectively enables the fluid from the primary flush fluid reservoir 1552 and the secondary fluid reservoir 1553 as described above.

[0072] When a primary flush fluid or a secondary fluid is required for cleaning the machine or the like, the connector 1514 and the tubing 1516 can be disconnected from the working fluid supply tank and connected to another mating connector 1518 that is connected to the fluid supply pump 1555. Other fluid supply systems of the present invention described herein may be used instead. When analyzing a sample, the working fluid and the sample fluid meet in the flow cell 1521, and the sample is hydrodynamically focused into the cuvette 1522 where the sample is investigated. After analysis, the mixed working fluid and sample can be considered as analyzer waste. This waste flows to the discharge waste accumulation section 1525 at atmospheric pressure. This accumulation section 1525 is at atmospheric pressure and maintains a constant pressure drop across the flow cell and the cuvette. One waste pump 1530 supplies a vacuum to draw all waste sources within the system. The analyzer waste within the discharge waste accumulation section 1525 is drawn to the waste management system 1527 through a tube having a known fluid resistance 1526. This resistor can be part of a tubing having a known length and inner diameter, an orifice, or the like. Any number of additional system waste sources 1528, such as system waste sources used to clean other parts of the machine, are further connected to the waste management system 1527 through a portion of tubing having a known fluid resistance 1529. The waste management system can be any fluid management system that combines multiple waste sources in parallel to a single output. One waste output of the waste management system 1527 is used to draw all waste sources through the waste pump 1530, and all of the system waste is stored in the waste tank 1531.

[0073] In one embodiment, the fluid management system comprises a particle sorter fluid management system belonging to a particle sorter configured to analyze and sort particles in a sample fluid. After analyzing the particles in the sample fluid, the analyzed sample fluid may flow from the output of the cuvette to a sorting block coupled to the output of the cuvette. The flow stream may exit from a nozzle within the sorting block having a nozzle diameter. To generate droplets, the nozzle may be rapidly vibrated by an acoustic device such as a piezoelectric element. To sort the particles in the sample, a droplet charging mechanism may charge the droplets of the flow stream containing the particle types to be sorted by charge, for example at the separation point of the flow stream. The droplets may pass through an electrostatic field and be deflected to one or more collection containers based on the polarity and magnitude of the charge of the droplets. Uncharged droplets are not deflected by the electrostatic field. The droplets may be collected in one or more sample collection containers or collection subsystems that are appropriately directed, for example arranged, to collect one or more flow streams generated by the deflection plates, whether charged or uncharged.

[0074] In such an embodiment, the fluid management system may be configured to analyze the sample fluid in the cuvette in the same manner as performed by the particle analyzer, as detailed above. For example, the system analyzes the detected light to measure the physical and fluorescence properties of the particles. The flow cytometer can further sort the particles based on these measured properties. The fluid management system for the particle sorter may be the same as the fluid management system for the particle analyzer, except that it may include a sorting block, a sorter waste fluid recovery subsystem (instead of the analyzer waste fluid recovery subsystem), and a sample recovery system having one or more sample recovery containers. The sorting block may be configured to be coupled, for example detachably, to the output of the cuvette. The waste fluid from sorting may be recovered by a sorter waste fluid recovery subsystem disposed in a droplet receiving relationship with the output of the sorting block. The waste fluid recovered by the sorter waste fluid recovery subsystem may be conveyed to a waste container via a waste management subsystem by a fluid transfer device. The waste fluid recovered by the sorter waste fluid recovery subsystem may be drawn into the waste container via the waste management subsystem by a vacuum generated by the fluid transfer device. The waste fluid from the sorting operation may include, for example, particles not sorted into the sample recovery container and the analyzed fluid (e.g., sample fluid, operating fluid).

[0075] As summarized above, in some cases, the fluid management system includes a sorting device waste fluid recovery subsystem configured to fluidly couple the output of a sorting block coupled to the output of the cuvette to a waste management subsystem. The sorting device waste fluid recovery subsystem may be configured to receive and recover waste fluid from the sorting operation by a flow-type particle analyzer. The sorting device waste fluid recovery subsystem may be configured to receive and recover waste fluid from the output of the sorting block, e.g., waste fluid that has flowed through the cuvette and exited the sorting block coupled to the cuvette. The sorting device waste fluid recovery subsystem may be disposed in a droplet receiving relationship with the sorting block. The sorting device waste fluid recovery subsystem may have any container and fluid line suitable for recovering waste fluid by sorting the sample.

[0076] In some cases, the sorting device waste fluid recovery subsystem has a waste aspirator for receiving waste from the output of the sorting block, and the waste aspirator is fluidly coupled to the waste management subsystem. The waste aspirator may have any component suitable for receiving and transporting waste fluid from the sorting block, such as tubes, containers, fluid lines, etc. The waste aspirator may transport the waste received from the sorting block to the waste management subsystem. In some cases, the waste fluid in the waste aspirator is drawn into the waste management subsystem by a vacuum generated by a fluid moving device fluidly coupled to the waste management subsystem. The waste aspirator may be configured to receive waste from the output of the sorting block. In some cases, the waste aspirator is disposed in a droplet receiving relationship with the output of the sorting block coupled to the cuvette. The waste aspirator may be fluidly coupled to the waste management subsystem with a known fluid resistance. In some cases, a resistor is provided in the fluid line coupling the waste aspirator to the waste management subsystem. The resistor may provide a known fluid resistance to the fluid line coupling the waste aspirator to the waste management subsystem. The resistor may be any resistor described herein.

[0077] In some cases, the fluid management system includes a sorting block (e.g., a detachable sorting block) configured to be removably coupled to the output of the cuvette. "Removably coupled" means that the sorting block can be coupled to, e.g., attached to, the cuvette and then later detached from, e.g., removed from, the cuvette at some point. 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 mounting component that can couple the sorting block 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 fluid subsystem connector. For example, the sorting block and the analyzer waste fluid recovery subsystem connector may both have the same manual connector configured to be removably coupled to the output of the cuvette.

[0078] The sorting block may have any suitable components that enable sorting of particles in the sample, such as sorting nozzles, and one or more deflection plates. To sort cells by the electrostatic method, the desired cells must be contained within charged droplets. To generate the droplets, the sorting block, e.g., the sorting nozzle, may vibrate rapidly by an acoustic device such as a piezoelectric element. The volume of the droplets may be estimated based on the hydrodynamic properties of the flow stream and the dimensions of the nozzle. To charge the droplets, the sorting block may have a charging element that can rapidly change the potential. Since the flow of cells may exit the sorting nozzle in a substantially downward vertical direction, after the droplets are formed, the droplets may also propagate in that direction. The droplets, whether charged or uncharged, may be appropriately directed, e.g., positioned, to be collected in a sample collection container or by the waste fluid recovery subsystem to collect one or more flow streams generated by the deflection plates. Methods and apparatuses for electrostatic cell sorting are described, for example, in U.S. Patent No. 9,952,076 and U.S. Patent No. 9,404,846, the entire disclosures of which are incorporated herein by reference.

[0079] The sorting block may have any suitable sorting nozzle configured to generate stream-in air droplets for sorting. The mixed working fluid and sample fluid may flow from the cuvette to the sorting nozzle and out into the flow stream through the orifice of the sorting nozzle. The orifice of the nozzle may have any diameter, for example 50 μm, 70 μm, 100 μm, or any other suitable diameter. The diameter of the nozzle may affect the characteristics of the flow stream, such as the dimensions of the flow stream, the droplet separation point, and the volume of the droplets. The flow stream may be a continuous flow of fluid or a series of droplets depending on the action of the droplet generator.

[0080] The sorting block may have any suitable droplet deflector configured to deflect particles in the flow stream flowing between the droplet deflectors. The flow stream may be a series of droplets that are partially deflected by a pair of deflection plates and split into multiple flows. Charged droplets may be deflected by the potential applied to the deflection plates and move away from the original path, for example towards a collection container. Droplets that are not charged, either positively or negatively, may not be deflected by the potential applied to the deflection plates and thus may proceed along the original path. In some cases, charged droplets are collected by the waste fluid recovery subsystem. In some cases, uncharged droplets are collected by the waste fluid recovery subsystem.

[0081] The sorting block may send the sorted particles to a sample recovery system or the sorting device waste fluid recovery subsystem. The sorting block may send the sorted sample, for example the sorted particles of interest, to a sample collection container. The collection container may be in a droplet receiving relationship with the sorting block. For example, the collection container may receive the cell product sorted by flow cytometry from the sorting block, for example in the form of droplets. In some cases, the sorting block sends the waste fluid to the waste aspirator of the sorting device waste fluid recovery subsystem. The waste aspirator may be in a droplet receiving relationship with the sorting block. For example, the waste aspirator may receive the waste fluid from the sorting block, for example in the form of droplets.

[0082] In some cases, the fluid management system may include a sample recovery system that includes a sample recovery container for receiving the selected sample from the output of the sorting block. The sample recovery container may be any suitable container (e.g., having a rigid or flexible wall) for receiving the selected particles. The sample recovery system may be configured to aseptically recover the selected particles. In some cases, the sample recovery system comprises, for example, any of the recovery systems described in US Patent Application Publication No. 2019 / 0331657, the entire disclosure of which is incorporated herein by reference.

[0083] FIG. 16 is a flowchart showing the main functions of a fluid management system configured to analyze and sort a sample according to an embodiment. The main functions are: 1601 supplying the working fluid (e.g., sheath) and the sample fluid to the flow cell at a controlled ratio; 1602 supplying any number of primary flush fluids and / or secondary fluids (e.g., for cleaning); 1603 hydrodynamically focusing the sample using the working fluid for sample interrogation through the cuvette; recovering the unselected waste fluid; 1605 managing and storing all system wastes (wastes from particle sorting 1604 and secondary system waste 1606). The dotted lines indicate that (a) the working fluid and the sample fluid may be supplied to the flow cell, or (b) any number of primary flush fluids or secondary system fluids may be supplied to the flow cell.

[0084] FIG. 17 is a flowchart showing the sub-functions within the main functions of a fluid management system configured to analyze and sort a sample according to an embodiment. The configuration is a configuration for managing the secondary system fluid and all waste fluids based on pumps. A fluid supply pump 1706 manages the primary flush system fluid and the secondary system fluid 1703. A waste pump 1714 manages all waste fluids from particle sorting 1712 and system waste 1713.

[0085] In the fluid configuration, the sheath and the sample are sent based on positive pressure, and a positive pressure source is used to control the flow rate ratio of the sheath / sample liquid. The working fluid (sheath) is controlled by the regulated air pressure 1702, and the sample fluid pressure is controlled by the regulated air pressure 1704. The air pressure feedback control 1701 is performed in a closed loop via inputs from the working fluid (sheath) chamber 1705 and the sample chamber 1707. Examples of feedback parameters include measurements such as the air pressure in the chamber and the liquid level height for pressure head correction. Further, the working fluid (sheath) pressure control signal is used as feedback for sample pressure control. The sample pressure setpoint is always the sheath pressure + the desired flow rate. This is because the sample pressure is always higher than the working fluid (sheath) pressure for the positive flow through the system. Therefore, it is possible to dynamically adjust the working fluid without changing the effective pressure drop of the sample fluid.

[0086] The pressurized working fluid (sheath) proceeds through the fluid supply system 1709 to the flow cell 1708, and the pressurized sample is supplied directly to the flow cell 1708. The fluid 1703 in any number of primary flush fluid reservoirs and secondary fluid reservoirs (for example, for the cleaning liquid) can be further sent to the fluid supply system 1709 via the fluid supply pump 1706 instead of the working fluid (sheath) 1705. The dotted lines indicate that (a) the working fluid may be supplied to the flow cell, or (b) any number of primary flush fluids or secondary system fluids may be supplied to the flow cell. This enables 100% flushing of the fluid system. The sample is investigated as the primary fluid and the sample fluid pass through the flow cell and the cuvette 1708.

[0087] For analysis, after the working fluid and the sample fluid flow through the flow cell and the cuvette, the fluid proceeds to the sorting nozzle 1710, which generates stream-in air droplets. These droplets can be sorted by the instrument sorting mechanism to the collection section 1711 or the waste 1712. All waste from the system (sorted waste 1712 and any secondary waste sources 1713 such as cleaning operations) are mixed and drawn out by one vacuum waste source such as the waste pump 1714. The mixed waste is sent to the waste storage section 1715.

[0088] Figure 18 is a schematic diagram showing the sorting device configuration of a flow cytometer according to an embodiment. A pressurized air source 1801 is used to supply air pressure to the electro-pneumatic pressure regulator. The pressure regulator 1802 controls the air pressure in the working fluid reservoir 1804, and the sample fluid regulator 1803 controls the pressure in the sample chamber 1805. A feedback control mechanism 1806 such as a PID control unit is used to transmit a control signal 1808 to the pressure regulator 1802. A similar feedback control mechanism 1807 is used to transmit a control signal 1809 to the sample fluid regulator 1803. The feedback of the working fluid pressure is obtained from the measured air pressure 1810 in the working fluid reservoir and the liquid level height 1813 in the working fluid reservoir measured using the liquid level measuring device 1812. The feedback of the sample fluid pressure is obtained from the measured air pressure 1811 in the sample chamber and the working fluid control signal 1808. The set value of the sample fluid is always controlled as a delta higher than the working fluid pressure so that the pressure drop in the sample line remains constant even when the working fluid pressure changes. The working fluid reservoir 1804 has a quick disconnect connection part 1815 that connects to the fitting connector 1814.

[0089] The fluid supply system of the present invention is further included. Any arbitrary amount of secondary fluid is stored in the secondary fluid reservoir 1853. In addition, a primary flash fluid reservoir 1852 is provided. When primary flash fluid or secondary fluid is required, for example, for cleaning the equipment, the fluid supply connector 1814 and the tubing 1816 can be disconnected from the working fluid supply tank and connected to another mating connector 1818 connected to the fluid supply pump 1855. Other fluid supply systems of the present invention described herein may be used instead. This pump can then supply fluid according to the settings of the flow control circuit 1854. The fluid is directed to the flow cell 1821. When analyzing a sample, the working fluid and the sample fluid meet in the flow cell 1821, and the sample is hydrodynamically focused into the cuvette 1822 where the sample is investigated. After analysis, the working fluid and the sample proceed through a sorting nozzle 1823 that generates stream-in air droplets for sorting. The sorted sample is collected in the sample collection section 1832, while the unsorted waste is collected in the sorted waste aspirator 1833. One waste pump 1830 supplies a vacuum to suck all waste sources within the system. The sorted waste in the sorted waste aspirator 1833 is drawn to the waste management system 1827 through a tube having a known fluid resistance 1834. This resistor can be a part of tubing having a known length and inner diameter, an orifice, etc. Any arbitrary number of additional system waste sources 1828, such as system waste sources used for cleaning other parts of the equipment, are further connected to the waste management system 1827 through a part of tubing having a known fluid resistance 1829. The waste management system can be any fluid management system that combines multiple waste sources in parallel to one output. One waste output of the waste management system 1827 is used to draw all waste sources through the waste pump 1830, and all of the system waste is stored in the waste tank 1831.

[0090] In one embodiment, the fluid management system comprises a hybrid system. The hybrid system may be configured to alternatively (a) analyze particles in a sample fluid (e.g., operate in an analyzer mode) according to any of the embodiments described herein, or (b) analyze and sort particles in a sample fluid according to any of the embodiments described herein (e.g., operate in a sorter mode). The hybrid system has an analyzer waste fluid recovery subsystem configured to fluidically couple the output of the cuvette to the waste management subsystem, and a sorter waste fluid recovery subsystem configured to fluidically couple the output of the sorting block coupled to the output of the cuvette to the waste management subsystem. The fluid management system may be configured such that, in some cases, (a) the analyzer waste fluid recovery subsystem fluidically couples the output of the cuvette to the waste management subsystem, or in other cases, (b) the sorter waste fluid recovery subsystem fluidically couples the output of the sorting block coupled to the output of the cuvette to the waste management subsystem. The fluid management system may be configured such that the user can select whether the analyzer waste fluid recovery subsystem fluidically couples the output of the cuvette to the waste management subsystem, or the sorter waste fluid recovery subsystem fluidically couples the output of the sorting block coupled to the output of the cuvette to the waste management subsystem. The user may select based on whether they desire (a) analysis of the sample, or (b) analysis and sorting of the sample by a flow-through particle analyzer. For example, to enable the analyzer mode, the user may manually couple the analyzer waste fluid recovery subsystem connector to the output of the cuvette, and the analyzer waste fluid recovery subsystem connector may fluidically couple the cuvette to the analyzer waste fluid recovery subsystem. In the analyzer mode, the sorting block may not be coupled to (or may be removed from) the output of the cuvette. To enable the sorter mode, the user may manually couple the sorting block to the output of the cuvette and position the sorter waste fluid recovery subsystem in a droplet receiving relationship with the output of the sorting block.In some cases, to switch from the analyzer mode to the sorter mode, the analyzer waste fluid recovery subsystem, such as the analyzer waste fluid recovery subsystem connector, is detached or removed from the cuvette, and then the sorting block is coupled to the cuvette. The hybrid fluid management system may comprise any of the elements of the particle analyzer fluid management system and the particle sorter fluid management system as described above.

[0091] FIG. 19 is a flowchart showing the main functions of a fluid management system configured to alternatively (a) analyze a sample or (b) analyze and sort a sample, according to an embodiment. The main functions are: 1901 supplying the working fluid (e.g., sheath) and the sample fluid to the flow cell at a controlled ratio; 1902 supplying any number of secondary system fluids (e.g., for washing); 1903 hydrodynamically focusing the sample using the working fluid for sample interrogation through the cuvette; 1904 recovering the analyzed sample as waste if the system is configured as an analyzer; 1905 recovering the unselected waste fluid if the system is configured as a sorter; and 1906 managing and storing all system wastes (wastes from particle analysis 1904, particle sorting 1905, and system waste 1907). The dotted lines before sample interrogation indicate that (a) the working fluid and the sample fluid may be supplied to the flow cell, or (b) any number of secondary system fluids may be supplied to the flow cell. The dotted lines after sample interrogation indicate that (a) the analyzed sample may be sorted, or (b) the waste from sample analysis may be recovered without sorting.

[0092] FIG. 20 is a flowchart showing sub - functions within the main functions of a fluid management system configured to alternatively (a) analyze a sample or (b) analyze and sort a sample. The configuration is one that manages secondary system fluids and all waste fluids based on pumps. A fluid supply pump 2006 manages primary flush system fluids and secondary system fluids 2003. A waste pump 2016 manages system waste 2015 in addition to waste fluids from a discharge waste accumulator 2013 and particle sorting 2014.

[0093] In the fluid configuration, the sheath and the sample are sent based on positive pressure, and a positive pressure source is used to control the flow rate ratio of the sheath / sample liquid. The working fluid (sheath) is controlled by an adjustable air pressure 2002, and the sample fluid pressure is controlled by an adjustable air pressure 2004. The air pressure feedback control 2001 is performed in a closed loop via inputs from the working fluid (sheath) chamber 2005 and the sample chamber 2007. Examples of feedback parameters include measurements such as the air pressure in the chamber and the liquid level height for pressure head correction. Further, the working fluid (sheath) pressure control signal is used as feedback for sample pressure control. The sample pressure setpoint is always the sheath pressure + the desired flow rate. This is because the sample pressure must always be higher than the working fluid (sheath) pressure for a positive flow through the system. Thus, it is possible to dynamically adjust the working fluid without changing the effective pressure drop of the sample fluid.

[0094] The pressurized working fluid (sheath) proceeds through the fluid supply system 2009 to the flow cell 2008, and the pressurized sample is supplied directly to the flow cell 2008. The fluid 2003 in any number of fluid supply tanks (e.g., for the cleaning fluid) can be further sent to the fluid supply system 2009 via the fluid supply pump 2006 instead of the working fluid (sheath) 2005. The dotted lines connecting to the fluid supply system indicate that (a) the working fluid may be supplied to the flow cell, or (b) any number of secondary system fluids may be supplied to the flow cell. Thus, 100% flushing of the fluid system becomes possible. When the working fluid and the sample fluid pass through the flow cell and the cuvette 2008, the sample is investigated.

[0095] The hybrid fluid configuration includes both the discharge waste accumulation section 2013 from the analyzer system and the sorting nozzle 2011, the sample collection section 2012, and the sorted waste collection section 2014 from the sorter configuration. The user can select either an operation mode of analyzing only the sample without the need for flow generation or analyzing and sorting the sample as it is in the sorter. When the system is configured as an analyzer, a waste connection 2010, such as a connector or fixture, may be coupled to the cuvette, and the waste connection fluidly couples the discharge waste accumulation section to the cuvette. When the system is configured as a sorter, the sorting nozzle 2011 may be coupled to the cuvette. The dotted lines after the sample investigation in the flow cell and the cuvette indicate that (a) the analyzed sample may be sorted (2011), or (b) the waste from the sample analysis is collected without sorting (2010). All waste from the system (analyzed waste (2013) or sorted waste (2014) and any secondary waste sources (2015) such as from the cleaning operation) are mixed and drawn out by one vacuum waste source such as the waste pump (2016). The mixed waste is sent to the waste storage section 2017.

[0096] FIG. 21 shows a hybrid configuration including a combination of both an analyzer configuration and a sorter configuration having components that enable switching between functions, according to one embodiment. A pressurized air source 2101 is used to supply air pressure to a pneumatic pressure regulator. The pressure regulator 2102 controls the air pressure within the working fluid reservoir 2104, and the sample fluid regulator 2103 controls the pressure within the sample chamber 2105. A feedback control mechanism 2106, such as a PID control unit, is used to transmit a control signal 2108 to the pressure regulator 2102. A similar feedback control mechanism 2107 is used to transmit a control signal 2109 to the sample fluid regulator 2103. The feedback of the working fluid pressure is obtained from the measured air pressure 2110 within the working fluid reservoir and the liquid level height 2113 within the working fluid reservoir measured using the liquid level measuring device 2112. The feedback of the sample fluid pressure is obtained from the measured air pressure 2111 within the sample chamber and the working fluid control signal 2108. The set value of the sample fluid is always controlled as a delta higher than the working fluid pressure so that the pressure drop across the sample line remains constant even when the working fluid pressure changes. The working fluid reservoir 2104 has a quick disconnect connection 2115 that connects to the fitting connector 2114.

[0097] The system further includes the fluid supply system of the present invention. Any arbitrary amount of secondary fluid is stored in the secondary fluid reservoir 2153. A primary flush fluid reservoir 2152 is further provided. When primary flush fluid or secondary fluid is required for equipment cleaning etc., the fluid supply connector 2114 and the tubing 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 may be used instead. This pump can then supply the primary flush fluid or the secondary fluid based on the settings of the flow control circuit 2154. The working fluid or the secondary fluid is directed to the flow cell 2121. When analyzing a sample, the working fluid and the sample fluid meet in the flow cell 2121, and the sample is hydrodynamically focused into the cuvette 2122 where the sample is investigated. After analysis, the working fluid and the sample proceed through a sorting nozzle 2123 that generates stream-in air droplets for sorting. The sorted sample is collected in the sample collection section 2132, while the unsorted waste is collected in the sorted waste aspirator 2133. If the user wants to operate the system in analyzer-only mode, the sorting nozzle 2123 is removed from the cuvette, and instead, a connector 2124 that connects to the atmosphere at atmospheric pressure is coupled to the cuvette at the discharge waste accumulation section 2125. This accumulation section 2125 is under atmospheric pressure and maintains a constant pressure drop across the flow cell and the cuvette. One waste pump 2130 supplies a vacuum to suck all waste sources within the system. The sorted waste in the sorted waste aspirator 2133 is drawn to the waste management system 2127 through a tube having a known fluid resistance 2134. The analyzer waste in the discharge waste accumulation section 2125 is drawn to the waste management system 2127 through a tube having a known fluid resistor 2126. These resistors can be part of tubing having a known length and inner diameter, an orifice, etc. Any arbitrary number of additional system waste sources 2128, such as system waste sources used to clean other parts of the equipment, are further connected to the waste management system 2127 through a portion of tubing having a known fluid resistance 2129.A waste management system can be any fluid management system that combines multiple waste sources in parallel into one output. One waste output of the waste management system 2127 is used to draw all waste sources through a waste pump 2130, and all of the system waste is stored in a waste tank 2131.

[0098] Flow cytometer Aspects of the present invention further include a flow cytometer comprising the fluid supply system of the present invention. The flow cytometer of interest, for example, comprises a flow cell configured to carry particles in a flow stream, a light source configured to irradiate the flow cell at an interrogation point, and the fluid supply system of the present invention. As described herein, a "flow cell" is described in its conventional meaning to refer to a component such as a cuvette that includes a flow path having a liquid flow stream for carrying particles within a sheath fluid. The cuvette of interest has a container through which the passageway passes. The flow stream may include a liquid sample injected from a sample tube. The flow cytometer of interest includes a flow path accessible to light. In some cases, the flow cell includes a transparent material (e.g., quartz) that allows light to pass through the flow cell. In some embodiments, the flow cell is a stream-in air flow cell in which optical interrogation of the particles is performed outside the flow cell (i.e., in free space).

[0099] In some cases, the frost stream is configured such that light from the light source irradiates at the investigation point. The frost stream in which the flow path is configured may include a liquid sample injected from a sample tube. In certain embodiments, the frost stream may include a narrow and rapidly flowing liquid stream in which linearly separated particles carried within the frost stream are arranged in a row and separated from each other. The "investigation point" described herein refers to, for example, a region within a flow cell where particles are irradiated by light from the light source. The size of the investigation point may vary as desired. For example, if 0 μm represents the axis of the light emitted by the light source, the investigation point may be within the range of -100 μm to 100 μm, for example -50 μm to 50 μm, for example -25 μm to 40 μm, for example -15 μm to 30 μm.

[0100] After irradiating the particles in the flow cell, the particle-modulated light may be observed. "Particle-modulated light" means the light received from the particles in the frost stream after irradiating the particles with light from the light source. In some cases, the particle-modulated light is side-scattered light. As described herein, side-scattered light refers to the light that is refracted and reflected by the surface and internal structure of the particles. In a further embodiment, the particle-modulated light includes forward-scattered light (i.e., light that mainly passes through the particles or travels around the particles in the forward direction). In still other cases, the particle-modulated light includes fluorescence (i.e., the light emitted from a fluorescent dye after irradiating with excitation wavelength light).

[0101] As described above, aspects of the present invention further include a light source configured to irradiate particles passing through the flow cell at the interrogation point. Any convenient light source may be used as the light source described herein. In some embodiments, the light source is a laser. In embodiments, the laser may be any convenient laser, such as a continuous wave laser. For example, the laser may 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 may be a helium neon (HeNe) laser. In some cases, the laser may be a gas laser, such as a helium neon laser, an argon laser, a krypton laser, a xenon laser, a nitrogen laser, a CO 2 laser, a CO laser, an argon fluoride (ArF) excimer laser, a krypton fluoride (KrF) excimer laser, a xenon chloride (XeCl) excimer laser, or a xenon fluoride (XeF) excimer laser, or a combination thereof. In other cases, the flow cytometer of interest comprises a dye laser, such as a stilbene laser, a coumarin laser, or a rhodamine laser. In still other cases, the laser of interest is 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 still other cases, the flow cytometer of interest comprises 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:YVO 4 laser, a Nd:YCa 4 O(BO 3 ) 3 laser, a Nd:YCOB laser, a titanium sapphire laser, a thulium YAG laser, a ytterbium YAG laser, a Yb 2 O 3 laser, or a cerium-doped laser, and combinations thereof.

[0102] The laser light source according to an embodiment may further include one or more optical adjustment components. In an embodiment, the optical adjustment component is disposed between the light source and the flow cell, and may include any device that can change the spatial width of the irradiation from the light source or other characteristics of the irradiation, 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 an embodiment, the flow cytometer of interest includes one or more focusing lenses. The focusing lens may be a reducing lens in one example. In still other embodiments, the flow cytometer of interest includes an optical fiber.

[0103] If the optical adjustment component is configured to move, the optical adjustment component may be configured to move continuously or at discrete intervals, such as increments of 0.01 μm or more, such as 0.05 μm or more, such as 0.1 μm or more, such as 0.5 μm or more, such as 1 μm or more, such as 10 μm or more, such as 100 μm or more, such as 500 μm or more, such as 1 mm or more, such as 5 mm or more, such as 10 mm or more, such as 25 mm or more.

[0104] Any movement protocol may be used to move the optical adjustment component structure, and the optical adjustment component structure is connected to, for example, a movable support base or directly connected to a device using a motor-driven translation stage, a lead screw translation assembly, a geared translation device, such as a motor type, particularly a stepping motor, a servo motor, a brushless electric motor, a brushed DC motor, a microstep drive motor, a high-resolution stepping motor.

[0105] The light source may be disposed at any suitable distance from the flow cell. For example, the light source and the flow cell may be separated by 0.005 mm or more, such as 0.01 mm or more, such as 0.05 mm or more, such as 0.1 mm or more, such as 0.5 mm or more, such as 1 mm or more, such as 5 mm or more, such as 10 mm or more, such as 25 mm or more, and for example, separated by a distance of 100 mm or more. Additionally, the light source may be disposed at any suitable angle with respect to the flow cell, for example, at an angle within the range of 10 degrees to 90 degrees, such as 15 degrees to 85 degrees, such as 20 degrees to 80 degrees, such as 25 degrees to 75 degrees, such as 30 degrees to 60 degrees, for example, at an angle of 90 degrees.

[0106] In some embodiments, the light source of interest comprises a plurality of lasers, such as 2 or more lasers, such as 3 or more lasers, such as 4 or more lasers, such as 5 or more lasers, such as 10 or more lasers, and for example, 15 or more lasers, configured to provide laser light for separate illumination of the flow stream. Depending on the desired wavelength of the light for illuminating the flow stream, each laser may have various specific wavelengths within the range of 200 nm to 1500 nm, such as 250 nm to 1250 nm, such as 300 nm to 1000 nm, such as 350 nm to 900 nm, such as 400 nm to 800 nm. In certain embodiments, one or more of a 405 nm laser, a 488 nm laser, a 561 nm laser, and a 635 nm laser may be included in the laser of interest.

[0107] As described above, the particle analyzer of interest may further include one or more particle-modulated light detectors for detecting particle-modulated light intensity data. In some embodiments, the one or more particle-modulated light detectors include one or more forward-scattered light detectors configured to detect forward-scattered light. For example, the particle analyzer of interest may include one forward-scattered light detector or multiple forward-scattered light detectors, such as two or more, such as three or more, such as four or more, such as five or more forward-scattered light detectors. In one embodiment, the particle analyzer has one forward-scattered light detector. In other embodiments, the particle analyzer has two forward-scattered light detectors.

[0108] Any convenient detector for detecting the collected light may be used for the forward-scattered light detectors described herein. Detectors of interest may include, but are not limited to, light sensors or photodetectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge-coupled devices (CCD), intensified charge-coupled devices (ICCD), light-emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMT), phototransistors, quantum dot photoconductors or photodiodes, and combinations thereof. In one embodiment, the collected light is measured with 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. In one embodiment, the detector is a photomultiplier tube, for example, having an active detection surface area in each region within the range of 0.01 cm 2 to 10 cm 2 , for example, 0.05 cm 2 to 9 cm 2 , for example, 0.1 cm 2 to 8 cm 2 , for example, 0.5 cm 2 to 7 cm 2 , for example, 1 cm 2 to 5 cm 2 .

[0109] In an embodiment, the forward scattered light detector is configured to measure light continuously or at separate intervals. In some cases, the detector of interest is configured to continuously measure the collected light. In other cases, the detector of interest is configured to measure at separate intervals, for example, every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds, for example, every 1000 milliseconds or at other intervals to measure light.

[0110] In a further embodiment, the one or more particle modulated light detectors may have one or more side scattered light detectors for detecting light at a side scattered wavelength (i.e., light refracted and reflected by the surface and internal structure of the particle). In some embodiments, the particle analyzer has one side scattered light detector. In other embodiments, the particle analyzer has a plurality of side scattered light detectors, for example, two or more, for example, three or more, for example, four or more, for example, five or more side scattered light detectors.

[0111] Any convenient detector for detecting the collected light may be used for the side scattered light detector described herein. Detectors of interest may include, but are not limited to, light sensors or photodetectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCD), ICCD (intensified charge-coupled device), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMT), phototransistors, quantum dot photoconductors or photodiodes, and combinations thereof. In one embodiment, the collected light is measured with 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. In one embodiment, the detector is a photomultiplier tube, for example, 0.01 cm 2 ~10 cm2 , for example, 0.05 cm 2 ~9 cm 2 , for example, 0.1 cm 2 ~8 cm 2 , for example, 0.5 cm 2 ~7 cm 2 , for example, 1 cm 2 ~5 cm 2 and is a photomultiplier tube having an active detection surface area in each region within the range of

[0112] In an embodiment, the particle analyzer to be targeted further has a fluorescence detector configured to detect light of one or more fluorescence wavelengths. In other embodiments, the particle analyzer has a plurality of fluorescence detectors, for example, two or more, for example, three or more, for example, four or more, five or more, ten or more, fifteen or more, for example, twenty or more fluorescence detectors.

[0113] Any convenient detector for detecting the collected light may be used for the fluorescence detector described herein. Detectors of interest may include, but are not limited to, light sensors or photodetectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge-coupled devices (CCD), ICCD (intensified charge-coupled device), light-emitting diodes, photon counters, bolometers, pyroelectric detectors, photoreistors, photovoltaic cells, photodiodes, photomultiplier tubes (PMT), phototransistors, quantum dot photoconductors or photodiodes, and combinations thereof. In certain embodiments, the collected light is measured with 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. In certain embodiments, the detector is a photomultiplier tube, for example, 0.01 cm 2 ~10 cm 2 , for example, 0.05 cm 2 ~9 cm 2 , for example, 0.1 cm 2 ~8 cm 2 , for example, 0.5 cm2 ~7 cm 2 、 for example, 1 cm 2 ~5 cm 2 and is a photomultiplier tube having an active detection surface area for each region within the range of.

[0114] When the particle analyzer of interest has a plurality of fluorescence detectors, each fluorescence detector may be the same, or the assembly of fluorescence detectors may be a combination of different types of detectors. For example, when the particle analyzer of interest has two fluorescence detectors, in some embodiments, the first fluorescence detector is a CCD type device and the second fluorescence detector (or image sensor) is a CMOS type device. In other embodiments, both the first fluorescence detector and the second fluorescence detector are CCD type devices. In still other embodiments, both the first fluorescence detector and the second fluorescence detector are CMOS type devices. In still other embodiments, the first fluorescence detector is a CCD type device and the second fluorescence detector is a photomultiplier tube (PMT). In still other embodiments, the first fluorescence detector is a CMOS type device and the second fluorescence detector is a photomultiplier tube. In still other embodiments, both the first fluorescence detector and the second fluorescence detector are photomultiplier tubes.

[0115] In embodiments of the present disclosure, the fluorescence detector of interest is configured to measure light collected at one or more wavelengths, such as two or more wavelengths, such as five or more different wavelengths, such as ten or more different wavelengths, such as twenty-five or more different wavelengths, such as fifty or more different wavelengths, such as one hundred or more different wavelengths, such as two hundred or more different wavelengths, such as three hundred or more different wavelengths, and for example, is configured to measure light emitted from a sample in a flow stream at four hundred or more different wavelengths. In some embodiments, two or more detectors of a particle analyzer as described herein are configured to measure light collected at the same wavelength or overlapping wavelengths.

[0116] In some embodiments, the fluorescence detector of interest is configured to measure light collected over a range of wavelengths (e.g., 200 nm to 1000 nm). In certain embodiments, the detector of interest is configured to collect a spectrum of light over a range of wavelengths. For example, the particle analyzer may have one or more detectors configured to collect a spectrum of light over one or more of the wavelength ranges from 200 nm to 1000 nm. In yet other embodiments, the detector of interest is configured to measure light emitted from a sample in a flow stream at one or more specific wavelengths. For example, the particle analyzer may have one or more detectors configured to measure light at one or more of 450 nm, 518 nm, 519 nm, 561 nm, 578 nm, 605 nm, 607 nm, 625 nm, 650 nm, 660 nm, 667 nm, 670 nm, 668 nm, 695 nm, 710 nm, 723 nm, 780 nm, 785 nm, 647 nm, 617 nm, and any combination thereof. In certain embodiments, one or more detectors may be configured to pair with a specific fluorophore, such as a fluorophore used with a sample in a fluorescence assay.

[0117] In some embodiments, the particle analyzer has one or more wavelength separators disposed between the flow cell and one or more particle-modulated light detectors. The term "wavelength separator" is used herein in its conventional sense to refer to an optical component configured to separate light collected from a sample into predetermined spectral regions. In some embodiments, the particle analyzer has one wavelength separator. In other embodiments, the particle analyzer has a plurality of wavelength separators, such as two or more wavelength separators, such as three or more, such as four or more, such as five or more, such as six or more, such as seven or more, such as eight or more, such as nine or more, such as ten or more, such as fifteen or more, such as twenty-five or more, such as fifty or more, such as seventy-five or more, such as one hundred or more wavelength separators. In some embodiments, the wavelength separator is configured to separate light collected from a sample into a predetermined spectral region by passing light having the predetermined spectral region and reflecting light in one or more remaining spectral regions. In other embodiments, the wavelength separator is configured to separate light collected from a sample into a predetermined spectral region by passing light having the predetermined spectral region and absorbing light in one or more remaining spectral regions. In still other embodiments, the wavelength separator is configured to spatially diffract light collected from a sample into a predetermined spectral region. Each wavelength separator may 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 one embodiment, the wavelength separator of the light detection system of interest is a dichroic mirror.

[0118] Suitable flow cytometry systems may include, but are not limited to, those described in Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford Univ. Press (1997); Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology No. 91, Humana Press (1997); Practical Flow Cytometry, 3rd ed., Wiley-Liss (1995); Virgo, et al. (2012) Ann Clin Biochem. Jan;49(pt 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, which disclosures are incorporated herein by reference.In some cases, the flow cytometry systems of interest include BD Biosciences FACSCanto™ flow cytometer, BD Biosciences FACSCanto™ II flow cytometer, BD Accuri™ flow cytometer, BD Accuri™ C6 Plus flow cytometer, BD Biosciences FACSCelesta™ flow cytometer, BD Biosciences FACSLyric™ flow cytometer, BD Biosciences FACSVerse™ flow cytometer, BD Biosciences FACSymphony™ flow cytometer, BD Biosciences LSRFortessa™ flow cytometer, BD Biosciences LSRFortessa™ X-20 flow cytometer, BD Biosciences FACSPresto™ flow cytometer, BD Biosciences FACSVia™ flow cytometer, and BD Biosciences FACSCalibur™ cell sorter, BD Biosciences FACSCount™ cell sorter, BD Biosciences FACSLyric™ cell sorter, BD Biosciences Via™ cell sorter, BD Biosciences Influx™ cell sorter, BD Biosciences Jazz™ cell sorter, BD Biosciences Aria™ cell sorter, BD Biosciences FACSAria™ II cell sorter, BD Biosciences FACSAria™ III cell sorter, BD Biosciences FACSAria™ Fusion cell sorter, BD Biosciences FACSMelody™ cell sorter, BD Biosciences FACSymphony™ S6 cell sorter, etc.

[0119] In some embodiments, the subject system is a flow cytometry system such as the flow cytometry system described in U.S. Patent No. 10,663,476, U.S. Patent No. 10,620,111, U.S. Patent No. 10,613,017, U.S. Patent No. 10,605,713, U.S. Patent No. 10,585,031, U.S. Patent No. 10,578,542, U.S. Patent No. 10,578,469, U.S. Patent No. 10,481,074, U.S. Patent No. 10,302,545, U.S. Patent No. 10,145,793, U.S. Patent No. 10,113,967, U.S. Patent No. 10,006,852, U.S. Patent No. 9,952,076, U.S. Patent No. 9,933,341, U.S. Patent No. 9,726,527, U.S. Patent No. 9,453,789, U.S. Patent No. 9,200,334, U.S. Patent No. 9,097,640, U.S. Patent No. 9,095,494, U.S. Patent No. 9,092,034, U.S. Patent No. 8,975,595, U.S. Patent No. 8,753,573, U.S. Patent No. 8,233,146, U.S. Patent No. 8,140,300, U.S. Patent No. 7,544,326, U.S. Patent No. 7,201,875, U.S. Patent No. 7,129,505, U.S. Patent No. 6,821,740, U.S. Patent No. 6,813,017, U.S. Patent No. 6,809,804, U.S. Patent No. 6,372,506, U.S. Patent No. 5,700,692, U.S. Patent No. 5,643,796, U.S. Patent No. 5,627,040, U.S. Patent No. 5,620,842, U.S. Patent No. 5,602,039, U.S. Patent No. 4,987,086, U.S. Patent No. 4,498,766, the entire disclosures of which are incorporated herein by reference.

[0120] In some cases, the flow cytometry system of the present invention is configured to image particles in a flow stream by fluorescence imaging with high-frequency tag emission (FIRE) as described in Diebold, et al. Nature Photonics Vol.7(10);806-810(2013) and U.S. Patent No. 9,423,353, U.S. Patent No. 9,784,661, U.S. Patent No. 9,983,132, U.S. Patent No. 10,006,852, U.S. Patent No. 10,078,045, U.S. Patent No. 10,036,699, U.S. Patent No. 10,222,316, U.S. Patent No. 10,288,546, U.S. Patent No. 10,324,019, U.S. Patent No. 10,408,758, U.S. Patent No. 10,451,538, U.S. Patent No. 10,620,111, U.S. Patent Application Publication No. 2017 / 0133857, U.S. Patent Application Publication No. 2017 / 0328826, U.S. Patent Application Publication No. 2017 / 0350803, U.S. Patent Application Publication No. 2018 / 0275042, U.S. Patent Application Publication No. 2019 / 0376895, and U.S. Patent Application Publication No. 2019 / 0376894, the disclosures of which are incorporated herein by reference.

[0121] FIG. 22 shows a system 2200 for flow cytometry according to an exemplary embodiment of the present invention. The system 2200 includes a flow cytometer 2210, a controller / processor 2290, and a memory 2295. The flow cytometer 2210 has 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 long pass (LP) filters 2255a-2255b, and one or more fluorescence detectors 2260a-2260f.

[0122] The excitation lasers 2215a to 2215c emit light in the form of laser beams. The wavelengths of the laser beams emitted from the excitation lasers 2215a to 2215c are 488 nm, 633 nm, and 325 nm, respectively, in the exemplary system of FIG. 22. The laser beams are first directed through one or more of beam splitters 2245a and 2245b. Beam splitter 2245a transmits light at 488 nm and reflects light at 633 nm. Beam splitter 2245b transmits UV light (light having wavelengths in the range of 10 to 400 nm) and reflects light at 488 nm and 633 nm.

[0123] Next, the laser beams are directed to a focusing lens 2220, which focuses the laser beams onto the portion of the fluid stream in the flow chamber 2225 where the particles of the sample are present. The flow chamber is part of a fluid system that guides the particles in the stream, usually one at a time, to the focused laser beam for investigation. The flow chamber can have a flow cell within a benchtop cytometer or a nozzle tip within a stream-in-air cytometer.

[0124] Light from one or more of the laser beams interacts with the particles in the sample by diffraction, refraction, reflection, scattering, and absorption, and is re-emitted at various different wavelengths depending on the characteristics of the particles, such as the size of the particles, the internal structure, and the presence of one or more fluorescent molecules attached to or naturally present on or within the particles. The fluorescent emission, as well as the diffracted, refracted, reflected, and scattered light, may be sent through one or more of beam splitters 2245c to 2245g, bandpass filters 2250a to 2250e, long-pass filters 2255a to 2255b, and fluorescence collection lens 2240 to one or more of a forward scatter detector 2230, a side scatter detector 2235, and one or more fluorescence detectors 2260a to 2260f.

[0125] The fluorescence collection lens 2240 collects the light emitted by the interaction between the particles and the laser beam and sends the light towards one or more beam splitters and filters. Band-pass filters such as band-pass filters 2250a to 2250e enable a narrow range of wavelengths to pass through the band-pass filter. For example, the band-pass filter 2250a is a 510 / 20 filter. The first number represents the center of the spectral band. The second number indicates the range of the spectral band. Therefore, the 510 / 20 filter extends 10 nm from the center of the spectral band to both sides, that is, from 500 nm to 520 nm. A short-pass filter transmits light with wavelengths below a specified wavelength. Long-pass filters such as long-pass filters 2255a to 2255b transmit light with wavelengths above a specified wavelength of the light. For example, the long-pass filter 2255b, which is a long-pass filter with a wavelength of 670 nm, transmits light with a wavelength of 670 nm or more. Filters are often selected to optimize the specificity of the detector for a particular fluorescent dye. The filter can be configured such that the spectral band of the light transmitted to the detector is close to the emission peak of the fluorescent dye.

[0126] The forward scatter detector 2230 is arranged slightly off-axis from the direct beam passing through the flow cell and is configured to detect diffracted light, that is, excitation light that mainly travels in the forward direction through or around the particles. The intensity of the light detected by the forward scatter detector depends on the overall size of the particles. The forward scatter detector may have a photodiode. The side scatter detector 2235 is configured to detect refracted and reflected light on the surface and internal structure of the particles, which tends to increase as the complexity of the particle structure increases. Fluorescent emission from fluorescent molecules bound to the particles can be detected by one or more fluorescence detectors 2260a to 2260f. The side scatter detector 2235 and the fluorescence detectors may have photomultiplier tubes. The signals detected by the forward scatter detector 2230, the side scatter detector 2235, and the fluorescence detectors can be converted into electronic signals (voltages) by the detectors. This data can provide information about the sample.

[0127] Those skilled in the art will recognize that the flow cytometer according to the embodiments of the present invention is not limited to the flow cytometer shown in FIG. 22 and 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 of various wavelengths and various different configurations.

[0128] During operation, the operation of the flow cytometer is controlled by a controller / processor 2290, and measurement data from the detector can be stored in a memory 2295 and processed by the controller / processor 2290. Although not explicitly shown, the controller / processor 2290 is connected to the detector to receive output signals from the detector and may be further connected to the electrical and electromechanical components of the flow cytometer 2210 to control lasers, fluid flow parameters, etc. An input / output (I / O) functional unit 2297 may be further provided in the system. The memory 2295, the controller / processor 2290, and the I / O functional unit 2297 may be provided as an integral part of the flow cytometer 2210. In such an embodiment, a display may further form part of the I / O functional unit 2297 to present experimental data to the user of the flow cytometer 2210. Alternatively, the memory 2295, the controller / processor 2290, and part or all of the I / O functional unit may be part of one or more external devices such as a general-purpose computer. In some embodiments, part or all of the memory 2295 and the controller / processor 2290 can communicate wirelessly or wired with the flow cytometer 2210. Together with the memory 2295 and the I / O functional unit 2297, the controller / processor 2290 may be configured to perform various functions related to the preparation and analysis of flow cytometer experiments.

[0129] The system illustrated in FIG. 22 includes six different detectors that detect fluorescence within six different wavelength bands, as defined by the configuration of filters and / or splitters in the beam path from the flow cell 2225 to each detector (which may be referred to herein as a "filter window" for a given detector). Different fluorescent molecules used in flow cytometry experiments emit light in their own characteristic wavelength bands. The particular fluorescent labels used in the experiment, and the associated fluorescence emission bands of the fluorescent labels, may be selected to generally match the detector's filter window. The I / O functional unit 2297 may be configured to receive data regarding a panel of fluorescent labels and a flow cytometry experiment having a plurality of cell populations, each having a subset of a plurality of markers. The I / O functional unit 2297 may be further configured to receive biological data that assigns one or more markers to one or more cell populations, marker concentration data, emission spectrum data, data that assigns labels to one or more markers, and cytometer configuration data. Flow cytometry experiment data, such as label spectral characteristics and flow cytometer configuration data, may be further stored in the memory 2295. The controller / processor 2290 may be configured to evaluate one or more assignments of labels to markers.

[0130] In some embodiments, the subject system is a particle sorting system configured to sort particles using a sealed particle sorting module as described in U.S. Patent Application Publication No. 2017 / 0299493, filed Mar. 28, 2017, the disclosure of which is incorporated herein by reference. In certain embodiments, particles of a sample (e.g., cells) are sorted using a sorting decision module having a plurality of sorting decision units as described in U.S. Patent Application Publication No. 2020 / 0256781, filed Dec. 23, 2019, the disclosure of which is incorporated herein by reference. In some embodiments, the system for sorting components of a sample comprises a particle sorting module having a deflector plate as described in U.S. Patent Application Publication No. 2017 / 0299493, filed Mar. 28, 2017, the disclosure of which is incorporated herein by reference.

[0131] FIG. 23 is a functional block diagram of an example of a sorting control system, such as a processor 2300, for analyzing and displaying a biological event. The processor 2300 can be configured to perform various processes for controlling a graphical display of a biological event.

[0132] A flow cytometer or sorting system 2302 can be configured to acquire biological event data. For example, a flow cytometer can generate flow cytometry event data (e.g., particle modulated light data). The flow cytometer 2302 can be configured to supply biological event data to the processor 2300. A data communication channel can be provided between the flow cytometer 2302 and the processor 2300. The biological event data can be supplied to the processor 2300 via the data communication channel.

[0133] 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 supply a graphical display including a first plot of the 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 plot. In some embodiments, the gate can be a logical combination of one or more illustrated regions of interest drawn on a histogram or bivariate plot of one parameter. In some embodiments, the display may be used to display particle parameters or saturation detector data.

[0134] Processor 2300 may be further configured to display the biological event data on display device 2306 within the gate as being different from other events within the biological event data outside the gate. For example, processor 2300 may be configured to render the color of the biological event data included within the gate as being different from the color of the biological event data outside the gate. Display device 2306 may be implemented as a monitor, tablet computer, smartphone, or other electronic device configured to provide a graphical interface.

[0135] Processor 2300 may be configured to receive a gate selection signal identifying a gate from a first input device. For example, the first input device may be implemented as a mouse 2310. The mouse 2310 can send a gate selection signal to the processor 2300 that identifies a gate displayed on the display device 2306 or operated via the display device 2306 (e.g., by clicking on or within a desired gate when the cursor is at the desired gate). In some embodiments, the first device may be implemented as a keyboard 2308 or as other means for supplying an input signal to the processor 2300, such as a touch screen, input pen, light detector, or voice recognition system. Some input devices may include multiple input functions. In such embodiments, each input function may be considered an input device. For example, as shown in FIG. 23, the mouse 2310 can include a right mouse button and a left mouse button, and the right mouse button and the left mouse button can each generate a trigger event.

[0136] Due to the trigger event, the processor 2300 can change how the data is displayed, which part of the data is actually displayed on the display device 2306, and / or provide an input to further processing such as the selection of a 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 change the visualization of the plot to facilitate gate processing.

[0137] Processor 2300 may be connected to a storage device 2304. The storage device 2304 may be configured to receive and store biological event data from the processor 2300. The storage device 2304 may be further configured to receive and store flow cytometry event data from the processor 2300. The storage device 2304 may be further configured to enable the processor 2300 to search for biological event data such as flow cytometry event data.

[0138] The display device 2306 may be configured to receive display data from the processor 2300. The display data may include a plot of the biological event data and a gate indicating an overview of the plot segments. The display device 2306 may be further configured to change the information displayed in response to inputs received from the processor 2300 in conjunction with inputs from the flow cytometer 2302, the memory device 2304, the keyboard 2308, and / or the mouse 2310.

[0139] In some embodiments, the processor 2300 can generate a user interface to receive exemplary events for sorting. For example, the user interface may include a mechanism for receiving exemplary events or exemplary images. The exemplary events or images, or exemplary gates, may be provided before collection of the event data for the sample, or based on the first set of events for a portion of the sample.

[0140] FIG. 24A is a schematic diagram showing a particle sorting system 2400 (e.g., a flow cytometer 2302) according to one embodiment presented herein. In some embodiments, the particle sorting system 2400 is a cell sorting system. As shown in FIG. 24A, a droplet formation transducer 2402 (e.g., a piezoelectric oscillator) is coupled to a fluid tube 2401, which may be coupled to, include, or be the nozzle 2403. Within the fluid tube 2401, sheath fluid 2404 hydrodynamically focuses the sample fluid 2406 containing particles 2409 to form a moving fluid column 2408 (e.g., a stream). Within the moving fluid column 2408, the particles 2409 (e.g., cells) are aligned in a row and cross a monitoring region 2411 (e.g., where a laser stream intersects) that is irradiated by an irradiation source 2412 (e.g., a laser). Due to the vibration of the droplet formation transducer 2402, the moving fluid column 2408 splits into a plurality of droplets 2410, and some of the droplets contain particles 2409.

[0141] During operation, a detection station 2414 (e.g., an event detector) identifies when a particle of interest (or a cell of interest) crosses the monitoring region 2411. The detection station 2414 supplies an input to a timing circuit 2428, which then supplies an input to a flash charge circuit 2430. At the droplet fission point notified by the timed droplet delay (Δt), flash charge can be applied to the moving fluid column 2408 so that the droplet of interest carries charge. The droplet of interest can contain one or more particles or cells to be sorted. Thereafter, the charged droplets can be sorted by activating a deflection plate (not shown) to deflect the charged droplets into a container such as a collection tube or a sample plate of a multi-well or micro-well, where the well or micro-well can be specifically associated with the droplet of interest. As shown in FIG. 24A, the droplets can be collected in a drain container 2438.

[0142] A detection system 2416 (e.g., a droplet boundary detector) serves to automatically determine the phase of the droplet drive signal when a particle of interest passes through the monitoring region 2411. An exemplary droplet boundary detector is described in U.S. Patent No. 7,679,039, the entire content of which is incorporated herein by reference. By the detection system 2416, the device can accurately calculate the position of each detected particle in the droplet. The detection system 2416 can supply an amplitude signal 2420 and / or a phase signal 2418, which are then supplied to an amplitude control circuit 2426 and / or a frequency control circuit 2424 (via an amplifier 2422). Thereafter, the amplitude control circuit 2426 and / or the frequency control circuit 2424 controls the droplet formation transducer 2402. The amplitude control circuit 2426 and / or the frequency control circuit 2424 can be provided within the control system.

[0143] In some embodiments, the sorting electronic devices (e.g., detection system 2416, detection station 2414, and processor 2440) can be coupled to a memory configured to store detected events and sorting decision results based on the detected events. The sorting decision results can be included in the event data regarding the particles. In some embodiments, the detection system 2416 and the detection station 2414 can be implemented as one detection unit or can be communicatively coupled such that event measurements are collected by one of the detection system 2416 or the detection station 2414 and provided to non-collecting elements.

[0144] FIG. 24B is a schematic diagram showing a particle sorting system according to one embodiment presented herein. The particle sorting system 2400 shown in FIG. 24B includes deflection plates 2452 and 2454. Charge can be applied through a stream charging wire in the barb. Thus, a stream of droplets 2410 containing particles 2409 is generated for analysis. The particles can be irradiated using one or more light sources (e.g., lasers) to generate light scattering information and fluorescence information. Information about the particles is analyzed by a sorting electronic device (not shown in FIG. 24B) or other detection system, etc. The deflection plates 2452 and 2454 can be independently controlled to attract or repel the charged droplets and direct the droplets towards a desired collection container (e.g., one of 2472, 2474, 2476, or 2478). As shown in FIG. 24B, the deflection plates 2452 and 2454 can be controlled to direct the particles along a first path 2462 towards container 2474 or along a second path 2468 towards container 2478. If the particle is not a particle of interest (e.g., does not exhibit scattering information or irradiation information within a specified sorting range), the deflection plate may allow the particle to continue to travel along flow path 2464. Such uncharged droplets may flow into a waste container via a suction device 2470 or the like.

[0145] The sorting electronic device may include starting to collect measurement values, receiving a fluorescence signal regarding the particles, and determining how to adjust the deflection plate to sort the particles. As an exemplary embodiment of the embodiment shown in FIG. 24B, a BD FACSAria (trademark) series flow cytometer commercially available from Becton, Dickinson and Company (Franklin Lakes, NJ) is included.

[0146] In some embodiments where the flow cytometer is a cell sorter, the flow cytometer is an image - compatible particle sorter. In such embodiments, the fluid supply system of the present invention may be used to supply the operating fluid, the primary flushing fluid, and / or the secondary fluid to the flow cell of the image - compatible particle sorter. In one embodiment, the system is fluorescence imaging using a high - frequency tag - emitting image - compatible particle sorter as shown in FIG. 25. The particle sorter 2500 includes an optical irradiation unit 2500a including a light source 2501 (e.g., a 488 nm laser), the light source 2501 generates an output beam of light 2501a, and the output beam 2501a is split into a beam 2502a and a beam 2502b by a beam splitter 2502. The optical beam 2502a is propagated 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 of light. In some cases, the output beam 2503a generated from the acousto - optic device 2503 includes a local oscillator beam and a plurality of high - frequency comb beams. The optical beam 2502b is propagated 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 of light. In some cases, the output beam 2504a generated from the acousto - optic device 2504 includes a local oscillator beam and a plurality of high - frequency comb beams. The output beam 2503a and the output beam 2504a respectively generated from the acousto - optic device 2503 and the acousto - optic device 2504 are combined by a beam splitter 2505 to generate an output beam 2505a, and the output beam 2505a is transmitted through an optical component 2506 (e.g., an objective lens) to irradiate the particles in the flow cell 2507. In one embodiment, the acousto - optic device 2503 (AOD) splits one laser beam into an array of beamlets each having a different optical frequency and angle. The second AOD 2504 adjusts the optical frequency of the reference beam, and the reference beam is then overlapped with the array of beamlets by a beam combiner 2505.In one embodiment, an optical irradiation system having a light source and an acousto-optic device may further include the optical irradiation systems described in Schraivogel, etal. (“High-speed fluorescence image-enabled cell sorting” Science (2022), 375 (6578): 315-320) and U.S. Patent Application Publication No. 2021 / 0404943, the disclosures of which are incorporated herein by reference.

[0147] The output beam 2505a irradiates the sample particles 2508 propagating through the flow cell 2507 (e.g., together with the sheath fluid 2509) in the irradiation region 2510. The sheath fluid 2509 may be supplied to the flow cell 2507 by the fluid supply system of the present invention, for example, as described above. As shown in the irradiation region 2510, a plurality of beams (e.g., the angular deflection high-frequency shift beams of light shown as dots across the irradiation region 2510) overlap with the reference local oscillation beam (shown as a shaded line across the irradiation region 2510). Since the overlapping beams have different optical frequencies, they exhibit beat behavior, and each beamlet transmits a sinusoidal modulation at a different frequency f 1-n with.

[0148] The light from the irradiated sample is transmitted to a light detection system 2500b having 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 further includes a bright field photodetector 2513 for generating a light loss image 2513a. In some embodiments, the forward scattered light photodetector 2511 and the side scattered light photodetector 2512 are photodiodes (e.g., avalanche photodiodes (APDs)). In some cases, the bright field photodetector 2513 is a photomultiplier tube (PMT). Fluorescence from the irradiated sample is further detected by fluorescence photodetectors 2514-2517. In some cases, the fluorescence photodetectors 2514-2517 are photomultiplier tubes. The light from the irradiated sample is guided to the side scattered light detection channel 2512 and the fluorescence detection channels 2514-2517 via a beam splitter 2520. The light detection system 2500b includes bandpass optical components 2521-2524 (e.g., dichroic mirrors) for propagating light of a predetermined wavelength to the photodetectors 2514-2517, respectively. In some cases, the optical component 2521 has a 534 nm / 40 nm bandpass. In some cases, the optical component 2522 has a 586 nm / 42 nm bandpass. In some cases, the optical component 2523 has a 700 nm / 54 nm bandpass. In some cases, the optical component 2524 has a 783 nm / 56 nm bandpass. The first number represents the center of the spectral band. The second number indicates the range of the spectral band. Thus, a 510 / 20 filter extends 10 nm from the center of the spectral band to both sides, i.e., from 500 nm to 520 nm.

[0149] Data signals generated according to the light detected by the forward scattered light detection channel 2511, the side scattered light detection channel 2512, the bright field light detection channel 2513, and the fluorescence detection channels 2514 to 2517 are processed by real-time digital processing by the processors 2550 and 2551. Based on the data signals generated by the processors 2550 and 2551, images 2511a to 2517a can be generated in each light detection channel. Image corresponding selection is performed according to the selection signal generated by the sorting trigger 2552. The sorting unit 2500c has a deflector plate 2531 for deflecting particles into the sample container 2532 or the waste stream 2533. In some cases, the sorting unit 2500c is configured to sort particles using a sealed particle sorting module as described in U.S. Patent Application Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosure of which is incorporated herein by reference. In one embodiment, the sorting unit 2500c has a sorting decision module having a plurality of sorting decision units as described in U.S. Patent Application Publication No. 2020 / 0256781, the disclosure of which is incorporated herein by reference.

[0150] FIG. 26 shows a general configuration of an exemplary computing device 2600 according to an embodiment. The general configuration of the computing device 2600 shown in FIG. 26 includes the arrangement of computer hardware and software components. However, not all of these general conventional elements are necessarily shown in order to provide an effective disclosure. As shown, the computing device 2600 includes a processing unit 2610, a network interface 2620, a computer-readable media drive 2630, an input / output device interface 2640, a display 2650, and an input device 2660, all of which may communicate with each other via a communication bus. The network interface 2620 may provide connectivity to one or more networks or computing systems. Accordingly, the processing unit 2610 may receive information and instructions from other computing systems or services via the network. The processing unit 2610 may further communicate with a memory 2670 and may further provide output information for any display 2650 via the input / output device interface 2640. For example, analysis software (e.g., data analysis software or program, such as FlowJo (registered trademark)) stored as executable instructions in the non-transitory memory of the analysis system may display flow cytometry event data to the user. The input / output device interface 2640 may further receive input from any input device 2660, such as a keyboard, mouse, digital pen, microphone, touch screen, gesture recognition system, voice recognition system, game pad, accelerometer, gyroscope, or other input device. Data may be further stored in a data storage unit 2690.

[0151] Method for analyzing a sample Aspects of the present invention further include a method of analyzing a sample. In the method of interest, (a) a sample is introduced into a device having a fluid supply system, and (b) an operating fluid is supplied to the device to analyze the sample. The fluid supply system for use in the method is described above and is configured to carry an operating fluid from a proximal end to a distal end and has an operating fluid line having an operating fluid reservoir connector at the proximal end configured to be fluidly coupled to an operating fluid reservoir containing the operating fluid, a primary flush fluid line configured to carry a primary flush fluid from a proximal end to a distal end and having a primary flush fluid reservoir connector at the proximal end configured to be fluidly coupled to a primary flush fluid reservoir containing the primary flush fluid, a secondary fluid line configured to carry a secondary fluid from a proximal end to a distal end and having a secondary fluid reservoir connector at the proximal end configured to be fluidly coupled to a secondary fluid reservoir containing the secondary fluid, a flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet, and one fluid connection configured to be fluidly coupled to the flow control circuit, the operating fluid line, and the fluid components of the device.

[0152] In some cases, the sample to be analyzed by the present method is a biological sample. The term "biological sample" is used in the conventional sense to refer to an entire organism, a plant, a fungus, or a subset of tissues, cells, or components of an animal, and the subset of tissues, cells, or components of an animal may, in some cases, be found in blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, amniotic fluid, umbilical cord blood of amniotic fluid, urine, vaginal fluid, or semen. Therefore, "biological sample" refers to both a natural organism or a subset of its tissues, and a homogenate, lysate, or extract prepared from a subset of an organism or its tissues, including, for example, plasma, serum, cerebrospinal fluid, lymph, skin sections, airways, gastrointestinal tract, cardiovascular system, urogenital tract, tears, saliva, milk, blood cells, tumors, organs, but not limited to these. The biological sample may be tissues of any type of organism, including both healthy tissues 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., and in some cases, the sample is a blood sample, including whole blood, such as blood obtained by venipuncture or finger prick (the blood may or may not be combined with any reagents, such as preservatives, anticoagulants, etc. prior to the assay).

[0153] In certain embodiments, the sample source is a "mammal" or "mammalian", and these terms are widely used to describe organisms belonging to the class Mammalia, 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 a human. The present method may be applied to samples obtained from human subjects of both genders and at any stage of development (i.e., neonates, infants, juveniles, adolescents, adults), and in certain embodiments, the human subject is a juvenile, adolescent, or adult. The present invention may be applied to samples from human subjects, but it should be understood that the method may also be further implemented for samples from other animal subjects (i.e., "non-human subjects"), including, but not limited to, birds, mice, rats, dogs, cats, livestock, and horses, etc.

[0154] The cell of interest may be an object characterized according to various parameters such as phenotypic characteristics identified by attaching a specific fluorescent label to the cell of interest. In some embodiments, the system is configured to deflect an analysis droplet determined to contain a target cell. Various cells may be characterized using the methods of the subject matter. 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 erythrocyte cells. Target cells of interest include cells having a cell surface marker or antigen that may be taken up or labeled by a simple affinity agent or conjugate thereof. For example, target cells 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 α / β, T cell receptor γ / δ, 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 HIV-containing cells, Treg cells, antigen-specific T cell populations, tumor cells, or hematopoietic progenitor cells (CD34+) from whole blood, bone marrow, or umbilical cord blood.

[0155] In the method of interest, particles may further be used in research, clinical trials, or treatment. In some embodiments, in the subject method, individual cells are obtained from a biological sample of a target fluid or tissue. For example, in the subject method, cells are obtained from a sample of a fluid or tissue used as a research specimen or diagnostic specimen for a disease such as cancer. Similarly, in the subject method, cells are obtained from a sample of a fluid or tissue used for treatment. A cell therapy protocol is a protocol that may, for example, prepare viable cell material including cells and tissues and introduce it into a subject as a therapeutic treatment. Symptoms that can be treated by administration of a sample sorted by flow cytometry include, but are not limited to, blood diseases, immune system disorders, organ damage, and the like.

[0156] General cell therapy protocols may include the steps of sample collection, cell isolation, genetic modification, culturing, in vitro expansion, cell harvesting, sample volume reduction, sample washing, biopreservation, storage, and introduction of cells into a subject. The protocol may be initiated by collecting viable cells and tissues from a tissue source of a subject to generate a sample of cells and / or tissues. The sample may be collected by any suitable procedure, including, for example, administration of a cell mobilizing agent to the subject, blood collection from the subject, removal of bone marrow from the subject, and the like. After the sample is collected, cell enrichment may be performed by a plurality of methods including, for example, a method based on centrifugation, a method based on filtration, sedimentation, magnetic separation, fluorescence-activated cell sorting (FACS), and the like. In some cases, the enriched cells may be genetically modified by any convenient method, such as gene editing via nuclease. The genetically modified cells can be cultured, activated, and expanded in vitro. In some cases, the cells are preserved, for example cryopreserved, and stored for future use. When used, the cells are thawed and then administered to a patient, for example, the cells may be injected into the patient.

[0157] Method of assembling a device Aspects of the present invention further include a method of assembling a device. In the method of interest, the fluid supply system of the present invention is operatively connected to a fluid component (e.g., a flow cell) of a device (e.g., a flow cytometer). The fluid supply system for use in the method is described above and includes a working fluid line having a working fluid reservoir connector at a proximal end configured to carry the working fluid from the proximal end to the distal end and fluidly coupled to a working fluid reservoir containing the working fluid, a primary flush fluid line having a primary flush fluid reservoir connector at a proximal end configured to carry the primary flush fluid from the proximal end to the distal end and fluidly coupled to a primary flush fluid reservoir containing the primary flush fluid, a secondary fluid line having a secondary fluid reservoir connector at a proximal end configured to carry the secondary fluid from the proximal end to the distal end and fluidly coupled to a secondary fluid reservoir containing the secondary fluid, a flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet, and one fluid connection configured to fluidly couple the flow control circuit, the working fluid line, and the fluid component of the device.

[0158] In some embodiments, a method fluidly couples a working fluid reservoir to a working fluid line. Optionally, the method further operably connects a pressurized air source to the working fluid reservoir. In some such cases, the method operably connects a pressure regulator to the working fluid reservoir and the pressurized air source. In certain cases, the method fluidly couples a primary flush fluid reservoir to a primary flush fluid line. In a particular embodiment, the method fluidly couples a secondary fluid reservoir to a secondary fluid line. The method may further fluidly couple a plurality of secondary fluid lines to a plurality of secondary fluid reservoirs. If the flow control circuit of the fluid supply system has a row of valves, in some embodiments according to the method, the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is farthest from the outlet of the flow control circuit. Optionally, the method operably connects a vacuum source to the fluid components of the device.

[0159] Utility The flow-type particle analyzer equipped with the fluid supply system of the present disclosure and the method of using the same are used in a variety of different applications including, for example, research and clinical applications. The flow-type particle analyzer may be used in applications where it is desirable to analyze and / or analyze and sort particle components in a sample, such as a biological sample. The fluid supply system of the present disclosure may be incorporated into any suitable flow-type particle system, such as a flow cytometer system. The fluid supply system may be used in any suitable application of a flow-type particle analyzer where a cleaning liquid flows through the flow-type particle analyzer to clean the fluid management system. The fluid supply system may be used in applications where it is not possible to access the space of a specific fluid path within the fluid management system and 100% sweeping cleaning of the fluid management system is desired. For example, a fluid management system that performs waste management control using one fluid transfer device may be used in applications where it is desirable to reduce the cost and complexity of manufacturing and operating a flow-type particle analyzer. A fluid management system, such as a hybrid system as described herein, may be further used in applications where an improvement in the adaptability of particle analysis is desired. Suitable applications include those where it is desirable to switch between (a) an analysis mode and (b) an analysis and sorting mode with a flow-type particle analyzer. Such applications may include, for example, cost-effective clinical or research applications where it is desirable to have the adaptability to use one flow cytometry system in the same laboratory or clinical site to (a) analyze particles or (b) analyze and sort particles.

[0160] The fluid supply system described herein may be used in a variety of different flow-type particle analyzers, such as flow cytometers. Suitable flow cytometry systems in which the subject fluid management system may be used include, but are not limited to, the flow cytometry systems described in U.S. Patent No. 9,952,076, U.S. Patent No. 9,933,341, U.S. Patent No. 9,726,527, U.S. Patent No. 9,453,789, U.S. Patent No. 9,200,334, U.S. Patent No. 9,097,640, U.S. Patent No. 9,095,494, U.S. Patent No. 9,092,034, U.S. Patent No. 8,975,595, U.S. Patent No. 8,753,573, U.S. Patent No. 8,233,146, U.S. Patent No. 8,140,300, U.S. Patent No. 7,544,326, U.S. Patent No. 7,201,875, U.S. Patent No. 7,129,505, U.S. Patent No. 6,821,740, U.S. Patent No. 6,813,017, U.S. Patent No. 6,809,804, U.S. Patent No. 6,372,506, U.S. Patent No. 5,700,692, U.S. Patent No. 5,643,796, U.S. Patent No. 5,627,040, U.S. Patent No. 5,620,842, U.S. Patent No. 5,602,039, the entire disclosures of which are incorporated herein by reference. In some cases, the flow cytometry systems of interest include, for example, the BD Biosciences FACSCanto(TM) II flow cytometer, the BD Accuri(TM) flow cytometer, the BD Biosciences FACSCelesta(TM) flow cytometer, the BD Biosciences FACSLyric(TM) flow cytometer, the BD Biosciences FACSVerse(TM) flow cytometer, the BD Biosciences FACSymphony(TM) flow cytometer, the BD Biosciences LSRFortessa(TM) flow cytometer, the BD Biosciences LSRFortess(TM) X-20 flow cytometer, and the like.

[0161] Additional suitable flow cytometry systems include, but are not limited to, the sorting flow cytometers described in U.S. Patent No. 3,960,449, U.S. Patent No. 4,347,935, U.S. Patent No. 4,667,830, U.S. Patent No. 4,704,891, U.S. Patent No. 4,770,992, U.S. Patent No. 5,030,002, U.S. Patent No. 5,040,890, U.S. Patent No. 5,047,321, U.S. Patent No. 5,245,318, U.S. Patent No. 5,317,162, U.S. Patent No. 5,464,581, U.S. Patent No. 5,483,469, U.S. Patent No. 5,602,039, U.S. Patent No. 5,620,842, U.S. Patent No. 5,627,040, U.S. Patent No. 5,643,796, U.S. Patent No. 5,700,692, U.S. Patent No. 6,372,506, U.S. Patent No. 6,809,804, U.S. Patent No. 6,813,017, U.S. Patent No. 6,821,740, U.S. Patent No. 7,129,505, U.S. Patent No. 7,201,875, U.S. Patent No. 7,544,326, U.S. Patent No. 8,140,300, U.S. Patent No. 8,233,146, U.S. Patent No. 8,753,573, U.S. Patent No. 8,975,595, U.S. Patent No. 9,092,034, U.S. Patent No. 9,095,494, and U.S. Patent No. 9,097,640, the entire disclosures of which are incorporated herein by reference. In some cases, the sorting flow cytometer is a cell sorter from Becton Dickinson, such as the BD Biosciences Influx™ cell sorter, the BD Biosciences FACSAria™ III cell sorter, the BD FACSAria™ Fusion cell sorter, the BD Biosciences FACSJazz™ cell sorter, the BD Biosciences FACSMelody™ cell sorter, and the like.

[0162] Notwithstanding the appended claims, the present disclosure is further defined by the following appendices.

[0163] An operating fluid line having at its proximal end an operating fluid reservoir connector configured to carry an operating fluid from the proximal end to the distal end and to be fluidly coupled to an operating fluid reservoir containing the operating fluid, A primary flush fluid line having at its proximal end a primary flush fluid reservoir connector configured to carry a primary flush fluid from the proximal end to the distal end and to be fluidly coupled to a primary flush fluid reservoir containing the primary flush fluid, A secondary fluid line having at its proximal end a secondary fluid reservoir connector configured to carry a secondary fluid from the proximal end to the distal end and to be fluidly coupled to a secondary fluid reservoir containing the secondary fluid, A flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively enable the primary flush fluid and the secondary fluid to pass through an outlet, One fluid connection configured to be fluidly coupled to the flow control circuit, the operating fluid line, and the fluid components of the device A fluid supply system comprising.

[0164] Appendix 2. The fluid supply system according to Appendix 1, wherein one fluid connection is constituted by a quick-connect fluid connector.

[0165] Appendix 3. The fluid supply system according to Appendix 1, wherein one fluid connection is constituted by a three-way valve.

[0166] Appendix 4. The fluid supply system according to Appendix 3, wherein the three-way valve has an internal volume within the range of 5 mm 3 ~100 mm 3 Appendix 5. The fluid supply system according to any one of Appendices 1 to 4, further comprising a fluid supply pump operably connected to the flow control circuit and one fluid connection.

[0167]

[0168] ​Appendix 6. The fluid supply system according to any one of Appendices 1 to 5, further comprising a working fluid reservoir fluidly connected to the working fluid line.

[0169] Appendix 7. The fluid supply system according to Appendix 6, wherein the working fluid is a sheath fluid.

[0170] Appendix 8. The fluid supply system according to Appendix 6 or 7, further comprising a pressurized air source operably connected to the working fluid reservoir.

[0171] Appendix 9. The fluid supply system according to Appendix 8, further comprising a pressure regulator operably connected to the working fluid reservoir and the pressurized air source.

[0172] Appendix 10. The fluid supply system according to any one of Appendices 1 to 9, further comprising a working fluid supply pump operably connected to one fluid connection portion.

[0173] Appendix 11. The fluid supply system according to any one of Appendices 1 to 10, further comprising a primary flush fluid reservoir fluidly connected to the primary flush fluid line.

[0174] Appendix 12. The fluid supply system according to Appendix 11, wherein the primary flush fluid is an inert fluid.

[0175] Appendix 13. The fluid supply system according to any one of Appendices 1 to 12, further comprising a secondary fluid reservoir fluidly connected to the secondary fluid line.

[0176] Appendix 14. The fluid supply system according to Appendix 13, wherein the secondary fluid is selected from a cleaning liquid and a reagent.

[0177] Appendix 15. The fluid supply system according to any one of Appendices 1 to 14, comprising a plurality of secondary fluid lines.

[0178] Appendix 16. The fluid supply system according to Appendix 15, wherein the number of the plurality of secondary fluid lines is in the range of 1 to 10.

[0179] Appendix 17. The fluid supply system according to any one of Appendices 1 to 16, wherein the flow control circuit has a row of valves.

[0180] Appendix 18. The fluid supply system according to Appendix 17, wherein the flow control circuit has a row of two-way valves.

[0181] Appendix 19. The fluid supply system according to Appendix 18, wherein the two-way valve is a stand-alone valve.

[0182] Appendix 20. The fluid supply system according to Appendix 18, wherein the two-way valve is attached to a manifold.

[0183] Appendix 21. The fluid supply system according to Appendix 17, wherein the flow control circuit has a row of three-way valves.

[0184] Appendix 22. The fluid supply system according to Appendix 21, wherein the three-way valve is a stand-alone valve.

[0185] Appendix 23. The fluid supply system according to Appendix 21, wherein the three-way valve is attached to a manifold.

[0186] Appendix 24. The fluid supply system according to any one of Appendices 17 to 23, wherein the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is furthest from the outlet of the flow control circuit.

[0187] Appendix 25. The fluid supply system according to any one of Appendices 1 to 16, wherein the flow control circuit has a multiport selection valve.

[0188] Appendix 26. The fluid supply system according to Appendix 25, wherein the multiport selection valve has an internal volume within the range of 5 mm 3 ~1000 mm 3

[0189] ​Appendix 27. The fluid supply system according to any one of Appendices 1 to 26, further comprising a device.

[0190] Appendix 28. The fluid supply system according to Appendix 27, wherein the device is a flow cytometer.

[0191] Appendix 29. The fluid supply system according to Appendix 28, wherein the fluid component of the device is a flow cell.

[0192] Appendix 30. The fluid supply system according to any one of Appendices 27 to 29, further comprising a vacuum source operably connected to the fluid component of the device.

[0193] Appendix 31. A method for analyzing a sample, (a) An operating fluid line having an operating fluid reservoir connector at a proximal end configured to carry an operating fluid from the proximal end to a distal end and fluidly coupled to an operating fluid reservoir containing the operating fluid, A primary flush fluid line having a primary flush fluid reservoir connector at a proximal end configured to carry a primary flush fluid from the proximal end to a distal end and fluidly coupled to a primary flush fluid reservoir containing the primary flush fluid, A secondary fluid line having a secondary fluid reservoir connector at a proximal end configured to carry a secondary fluid from the proximal end to a distal end and fluidly coupled to a secondary fluid reservoir containing the secondary fluid, A flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through an outlet, One fluid connection configured to fluidly couple the flow control circuit, the operating fluid line, and the fluid component of the device Introducing a sample into a device comprising a fluid supply system having the above components, (b) Supplying the operating fluid to the device to analyze the sample.

[0194] Appendix 32. The method according to Appendix 31, wherein one fluid connection part is constituted by a quick-connect fluid connector.

[0195] Appendix 33. The method according to Appendix 31, wherein one fluid connection part is constituted by a three-way valve.

[0196] Appendix 34. The three-way valve has an internal volume within the range of 5 mm 3 to 100 mm 3 The method according to Appendix 33.

[0197] Appendix 35. The fluid supply system further has a flow control circuit and a fluid supply pump operably connected to one fluid connection part. The method according to any one of Appendices 31 to 34.

[0198] Appendix 36. The fluid supply system further has a working fluid reservoir fluidly coupled to the working fluid line. The method according to any one of Appendices 31 to 35.

[0199] Appendix 37. The working fluid is a sheath fluid. The method according to Appendix 36.

[0200] Appendix 38. The fluid supply system further has a pressurized air source operably connected to the working fluid reservoir. The method according to Appendix 36 or 37.

[0201] Appendix 39. The flow control system further has a pressure regulator operably connected to the working fluid reservoir and the pressurized air source. The method according to Appendix 38.

[0202] Appendix 40. The fluid supply system further has a working fluid supply pump operably connected to one fluid connection part. The method according to any one of Appendices 31 to 39.

[0203] Appendix 41. The fluid supply system further has a primary flush fluid reservoir fluidly coupled to the primary flush fluid line. The method according to any one of Appendices 31 to 40.

[0204] Supplementary Note 42. The method according to Supplementary Note 41, wherein the primary flash fluid is an inert fluid.

[0205] Supplementary Note 43. The method according to any one of Supplementary Notes 31 to 42, wherein the fluid supply system further has a secondary fluid reservoir fluidly connected to the secondary fluid line.

[0206] Supplementary Note 44. The method according to Supplementary Note 43, wherein the secondary fluid is selected from a cleaning liquid and a reagent.

[0207] Supplementary Note 45. The method according to any one of Supplementary Notes 31 to 44, wherein the fluid supply system has a plurality of secondary fluid lines.

[0208] Supplementary Note 46. The method according to Supplementary Note 45, wherein the number of the plurality of secondary fluid lines is in the range of 1 to 10.

[0209] Supplementary Note 47. The method according to any one of Supplementary Notes 31 to 46, wherein the flow control circuit has a row of valves.

[0210] Supplementary Note 48. The method according to Supplementary Note 47, wherein the flow control circuit has a row of two-way valves.

[0211] Supplementary Note 49. The method according to Supplementary Note 48, wherein the two-way valve is a stand-alone valve.

[0212] Supplementary Note 50. The method according to Supplementary Note 48, wherein the two-way valve is attached to a manifold.

[0213] Supplementary Note 51. The method according to Supplementary Note 47, wherein the flow control circuit has a row of three-way valves.

[0214] Supplementary Note 52. The method according to Supplementary Note 51, wherein the three-way valve is a stand-alone valve.

[0215] Supplementary Note 53. The method according to Supplementary Note 51, wherein the three-way valve is attached to a manifold.

[0216] Supplement 54. The method according to any one of Supplements 47 to 53, wherein the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is furthest from the outlet of the flow control circuit.

[0217] Supplement 55. The method according to any one of Supplements 31 to 46, wherein the flow control circuit has a multi-port selection valve.

[0218] Supplement 56. The method according to Supplement 55, wherein the multi-port selection valve has an internal volume within the range of 5 mm 3 ~1000 mm 3 Supplement 57. The method according to any one of Supplements 31 to 56, wherein the device is a flow cytometer.

[0219] Supplement 58. The method according to Supplement 57, wherein the fluid component of the device is a flow cell.

[0220] Supplement 59. The method according to Supplement 57 or 58, wherein the fluid supply system has a vacuum source operably connected to the fluid component of the device.

[0221] Supplement 60. The method according to any one of Supplements 31 to 59, wherein the sample is a biological sample.

[0222] Supplement 61. A method of assembling a device,

[0223] an actuating fluid line having a proximal end with an actuating fluid reservoir connector configured to be fluidly coupled to an actuating fluid reservoir containing an actuating fluid and configured to carry the actuating fluid from the proximal end to the distal end through the actuating fluid; and a primary flush fluid line having a proximal end with a primary flush fluid reservoir connector configured to be fluidly coupled to a primary flush fluid reservoir containing a primary flush fluid and configured to carry the primary flush fluid from the proximal end to the distal end through the primary flush fluid; and a primary flush fluid line having a proximal end with a primary flush fluid reservoir connector configured to be fluidly coupled to a primary flush fluid reservoir containing a primary flush fluid and configured to carry the primary flush fluid from the proximal end to the distal end through the primary flush fluid; and A secondary fluid line having a proximal end with a secondary fluid reservoir connector configured to carry the secondary fluid from the proximal end to the distal end and fluidly coupled to a secondary fluid reservoir containing the secondary fluid, A flow control circuit fluidly connected to the distal ends of the primary flush fluid line and the secondary fluid line and configured to selectively allow the primary flush fluid and the secondary fluid to pass through the outlet, One fluid connection configured to fluidly couple the flow control circuit, the working fluid line, and the fluid components of the device A method of operably connecting a fluid supply system comprising: to the fluid components of a device.

[0224] Appendix 62. The method according to Appendix 61, wherein one fluid connection is constituted by a quick-connect fluid connector.

[0225] Appendix 63. The method according to Appendix 61, wherein one fluid connection is constituted by a three-way valve.

[0226] Appendix 64. The method according to Appendix 63, wherein the three-way valve has an internal volume within the range of 5 mm 3 ~100 mm 3 The method according to Appendix 63, having an internal volume within the range of.

[0227] Appendix 65. The method according to any one of Appendices 61 to 64, wherein the fluid supply system further has a fluid supply pump operably connected to the flow control circuit and one fluid connection.

[0228] Appendix 66. The method according to any one of Appendices 61 to 65, wherein an operating fluid reservoir is fluidly coupled to the operating fluid line.

[0229] Appendix 67. The method according to Appendix 66, wherein the operating fluid is a sheath fluid.

[0230] Appendix 68. The method according to Appendix 66 or 67, wherein a pressurized air source is operably connected to the operating fluid reservoir.

[0231] Appendix 69. The method according to Appendix 68, wherein the pressure regulator is operably connected to a working fluid reservoir and a pressurized air source.

[0232] Appendix 70. The method according to any one of Appendices 61 - 69, wherein the fluid supply system further has a working fluid supply pump operably connected to one fluid connection.

[0233] Appendix 71. The method according to any one of Appendices 61 - 70, wherein the primary flash fluid reservoir is fluidly coupled to the primary flash fluid line.

[0234] Appendix 72. The method according to Appendix 71, wherein the primary flash fluid is an inert fluid.

[0235] Appendix 73. The method according to any one of Appendices 61 - 72, wherein the secondary fluid reservoir is fluidly coupled to the secondary fluid line.

[0236] Appendix 74. The method according to Appendix 73, wherein the secondary fluid is selected from a cleaning liquid and a reagent.

[0237] Appendix 75. The method according to any one of Appendices 61 - 74, wherein the fluid supply system has a plurality of secondary fluid lines.

[0238] Appendix 76. The method according to Appendix 75, wherein the number of the plurality of secondary fluid lines is in the range of 1 - 10.

[0239] Appendix 77. The method according to any one of Appendices 61 - 76, wherein the flow control circuit has a row of valves.

[0240] Appendix 78. The method according to Appendix 77, wherein the flow control circuit has a row of two - way valves.

[0241] Appendix 79. The method according to Appendix 78, wherein the two - way valve is a stand - alone valve.

[0242] Appendix 80. The method according to Appendix 78, wherein the two - way valve is attached to a manifold.

[0243] Appendix 81. The method according to Appendix 77, wherein the flow control circuit has a single row of three-way valves.

[0244] Appendix 82. The method according to Appendix 81, wherein the three-way valve is a stand-alone valve.

[0245] Appendix 83. The method according to Appendix 81, wherein the three-way valve is attached to a manifold.

[0246] Appendix 84. The method according to any one of Appendices 77 to 83, wherein the distal end of the primary flush fluid line is fluidly coupled to a valve within the row of valves that is furthest from the outlet of the flow control circuit.

[0247] Appendix 85. The method according to any one of Appendices 61 to 76, wherein the flow control circuit has a multiport selection valve.

[0248] Appendix 86. The method according to Appendix 85, wherein the multiport selection valve has an internal volume within the range of 5 mm 3 to 1000 mm 3 .

[0249] Appendix 87. The method according to any one of Appendices 61 to 86, wherein the device is a flow cytometer.

[0250] Appendix 88. The method according to Appendix 87, wherein the fluid component of the device is a flow cell.

[0251] Appendix 89. The method according to Appendix 87 or 88, wherein a vacuum source is operably connected to the fluid component of the device.

[0252] The above invention has been described in some detail by way of illustration and example for ease of understanding. However, it will be readily apparent to those skilled in the art that, in light of the teachings of the present invention, some changes and modifications may be made without departing from the spirit and scope of the appended claims.

[0253] Therefore, the foregoing is only illustrative of the essence of the present invention. It is obvious that those skilled in the art can devise various configurations that embody the essence of the present invention and are included within the spirit and scope of the present invention, although not explicitly described or shown in this specification. Furthermore, all examples and conditional terms described in this specification are essentially intended to assist the reader in understanding the essence of the present invention and the concepts given by the inventor for the advancement of the present technical field, and should not be construed as being limited to the specifically described examples and conditions. Moreover, all descriptions in this specification that describe the essence, aspects, and embodiments of the present invention as well as specific examples of the present invention are intended to include both structural and functional equivalents of the present invention. In addition, such equivalents are intended to include both currently known equivalents and equivalents developed in the future, that is, all developed elements that perform the same function regardless of their structure. Furthermore, such disclosure is not generally intended to be published, whether or not explicitly recited in the claims.

[0254] Therefore, it is intended that the scope of the present invention not be limited to the exemplary embodiments presented and described in this specification. Rather, the scope and spirit of the present invention are embodied by the appended claims. With respect to the claims, 35 U.S.C. § 112(f) or 35 U.S.C. § 112, ¶ 6 is clearly defined to be applicable only to limitations in a claim when the exact phrase "means for" or "step for" begins the limitation in the claim, and 35 U.S.C. § 112(f) or 35 U.S.C. § 112, ¶ 6 is not applicable when such exact phrase is not used in the claim limitation.

[0255] Cross - reference to related applications This application claims the benefit of the filing date of U.S. Provisional Patent Application No. 63 / 546,024, filed on October 27, 2023, under 35 U.S.C. § 119(e), the entire disclosure of which is incorporated herein by reference.

Claims

1. an actuation fluid line configured to carry an actuation fluid from a proximal end to a distal end, the actuation fluid line having an actuation fluid reservoir connector at the proximal end configured to fluidly couple to an actuation fluid reservoir containing the actuation fluid; a primary flush fluid line configured to carry a primary flush fluid from a proximal end to a distal end, the primary flush fluid line having 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; a secondary fluid line configured to carry a secondary fluid from a proximal end to a distal end, the secondary fluid line having a secondary fluid reservoir connector at the proximal end configured to fluidly couple to a secondary fluid reservoir containing the secondary fluid; 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 the primary flush fluid and the secondary fluid to pass through an outlet; a fluid connection configured to fluidly couple to the flow control circuit, the actuation fluid line, and a fluid component of an instrument; A fluid delivery system comprising:

2. 2. The fluid delivery system of claim 1, wherein the one fluid connection comprises a quick connect fluid connector.

3. 2. The fluid supply system of claim 1, wherein the one fluid connection comprises a three-way valve.

4. The fluid supply system of any one of claims 1 to 3, further comprising a fluid supply pump operatively connected to said flow control circuit and said one fluid connection.

5. The fluid supply system of any one of claims 1 to 4, further comprising an actuating fluid reservoir fluidly coupled to the actuating fluid line.

6. A fluid supply system according to any one of claims 1 to 5, further comprising an actuation fluid supply pump operatively connected to said one fluid connection.

7. The fluid supply system of any one of claims 1 to 6, further comprising a primary flush fluid reservoir fluidly coupled to the primary flush fluid line.

8. The fluid supply system of any one of claims 1 to 7, further comprising a secondary fluid reservoir fluidly coupled to the secondary fluid line.

9. A fluid supply system according to any one of the preceding claims, comprising a plurality of secondary fluid lines.

10. A fluid delivery system according to any preceding claim, wherein the flow control circuit comprises an in-line valve.

11. The fluid supply system of any one of claims 1 to 11, wherein the flow control circuit comprises a multi-port selection valve.

12. The fluid delivery system of any one of claims 1 to 11, further comprising a flow cytometer.

13. 1. A method for analyzing a sample, comprising the steps of: (a) an actuation fluid line configured to carry an actuation fluid from a proximal end to a distal end, the actuation fluid line having an actuation fluid reservoir connector at the proximal end configured to fluidly couple to an actuation fluid reservoir containing the actuation fluid; a primary flush fluid line configured to carry a primary flush fluid from a proximal end to a distal end, the primary flush fluid line having 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; a secondary fluid line configured to carry a secondary fluid from a proximal end to a distal end, the secondary fluid line having a secondary fluid reservoir connector at the proximal end configured to fluidly couple to a secondary fluid reservoir containing the secondary fluid; 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 the primary flush fluid and the secondary fluid to pass through an outlet; a fluid connection configured to fluidly couple to the flow control circuit, the actuation fluid line, and a fluid component of an instrument; introducing the sample into an instrument having a fluid delivery system having a (b) supplying a working fluid to said instrument to analyze said sample.

14. The method according to claim 13, wherein the flow control circuit is a flow control circuit as defined in any one of claims 2 to 12.

15. A method of assembling a device, comprising: an actuation fluid line configured to carry an actuation fluid from a proximal end to a distal end, the actuation fluid line having an actuation fluid reservoir connector at the proximal end configured to fluidly couple to an actuation fluid reservoir containing the actuation fluid; a primary flush fluid line configured to carry a primary flush fluid from a proximal end to a distal end, the primary flush fluid line having 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; a secondary fluid line configured to carry a secondary fluid from a proximal end to a distal end, the secondary fluid line having a secondary fluid reservoir connector at the proximal end configured to fluidly couple to a secondary fluid reservoir containing the secondary fluid; 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 the primary flush fluid and the secondary fluid to pass through an outlet; a fluid connection configured to fluidly couple to the flow control circuit, the actuation fluid line, and a fluid component of an instrument; operatively connecting a fluid delivery system comprising: a fluid component of an instrument;

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