Particle sorting module with alignment window, system and method of use thereof

Abstract

Aspects of the present disclosure include a particle sorting module having an opening configured to visualize droplets of a deflected fluid stream. A particle sorting module according to certain embodiments includes a housing having a proximal end, a distal end, and a wall therebetween, the wall having an opening positioned in the wall, the opening being configured to align the fluid stream with one or more sample containers at the distal end. Systems and methods for aligning a fluid stream with one or more sample containers and sorting particles of a sample (e.g., a biological sample) are also provided. A kit having one or more particle sorting modules is also described, the particle sorting modules being suitable for coupling with a particle sorting system and being suitable for implementing the subject method.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority under 35 USC §119(e) to U.S. Provisional Patent Application Serial No. 62 / 752,793, filed on October 30, 2018; the disclosure of which is incorporated herein by reference. Background Art

[0003] Flow particle sorting systems (e.g., sorting flow cytometers) are used to sort particles in a fluid sample based on at least one measured characteristic of the particles. In a flow particle sorting system, particles (e.g., molecules, analyte-bound beads, or single cells) in a fluid suspension pass in a flow through a detection region where a sensor detects particles contained in the flow of the type to be sorted. When a particle of the type to be sorted is detected, the sensor triggers a sorting mechanism that selectively isolates the particle of interest.

[0004] Particle sensing is typically performed by passing a fluid stream through a detection region where the particles are exposed to illumination light from one or more lasers and the light scattering and fluorescence properties of the particles are measured. Particles or their components may be labeled with fluorescent dyes to facilitate detection, and multiple different particles or components may be detected simultaneously by labeling the different particles or components with spectrally distinct fluorescent dyes. Detection is performed using one or more light sensors to facilitate independent measurement of the fluorescence of each different fluorescent dye.

[0005] In order to sort the particles in the sample, the droplet charging device charges the droplets of the fluid stream containing the type of particles to be sorted with an electric charge at the demarcation point of the fluid stream. The droplets pass through the electrostatic field and are deflected into one or more collection containers based on the polarity and magnitude of the charge on the droplets. Droplets that do not carry a charge are not deflected by the electrostatic field. Summary of the invention

[0006] Aspects of the present disclosure include a particle sorting module having an opening configured to visualize droplets of a deflected fluid stream. A particle sorting module according to certain embodiments includes a housing having a proximal end, a distal end, and a wall therebetween, the wall having an opening positioned in the wall, the opening being configured to align the fluid stream with one or more sample containers at the distal end. In some embodiments, the opening is located in a wall downstream of a droplet deflector that deflects droplets of the fluid stream into one or more sample containers. In some cases, the opening is positioned 0.5 cm to 5 cm from the droplet deflector. In other cases, the opening is positioned 0.5 cm to 5 cm from the sample container at the distal end of the particle sorting module. In some embodiments, the opening includes an optical adjustment component, such as a focusing lens, for propagating light from an irradiation source to the deflected fluid stream. In other embodiments, the optical adjustment component is a transparent optical window. In some cases, the transparent optical window may include one or more reference identifiers, such as one or more marks associated with the boundaries of one or more sample containers at the distal end of the housing. In some cases, the reference identifier includes a mark indicating the center of the distal end of the housing. In some cases, the particle sorting module includes a waste container and two sample containers coupled to the distal end. In some embodiments, the transparent optical window includes a mark associated with the boundaries of the waste container and the two sample containers. In certain embodiments, the particle sorting module of interest includes a fluid pool nozzle located at the proximal end of the housing and a sample interrogation area connected to the fluid pool nozzle fluid. In some cases, the sample interrogation area has a test tube (e.g., a glass or plastic test tube). In some particle sorting modules, the test tube is co-molded. In other particle sorting modules, an adhesive is adhered to the test tube. In other particle sorting modules, the test tube is integral with the particle sorting module. The particle sorting module may also include one or more of a sample inlet and a sheath fluid inlet at the proximal end.

[0007] Aspects of the present disclosure also include systems for aligning a fluid stream with one or more sample containers and sorting particles of a sample (e.g., a biological sample). A system according to certain embodiments includes a particle sorting module having an opening and an imaging sensor, the opening being used to visualize droplets deflected from the flow, for example as described above, the imaging sensor being configured to capture one or more images of the deflected fluid stream passing through the opening. In some embodiments, the system includes a laser for irradiating the fluid stream passing through the opening. The opening may include one or more optical adjustment components, such as a focusing lens or a transparent optical window. In certain embodiments, the system includes a processor, the processor including a memory coupled to the processor in an operative manner, wherein the memory includes instructions stored thereon, the instructions when executed by the processor causing the processor to capture an image of the opening of the particle sorting module to obtain an imaging opening; and calculating the pixel position of the boundary of each port at the distal end of the housing along the horizontal axis in the imaging opening. In these embodiments, calculating the pixel position may include calculating the total number of pixels passing through the imaging opening along the horizontal axis; and multiplying the total number of pixels by a predetermined ratio. In some cases, the predetermined ratio is calculated by dividing the number of pixels of each port boundary in the imaging opening by the total number of pixels passing through the imaging opening along the horizontal axis. In some embodiments, the system includes a memory having instructions stored thereon, which, when executed by a processor, cause the processor to align the fluid flow with a port at the distal end of the housing. In other embodiments, the system includes a memory having instructions stored thereon, which, when executed by the processor, cause the processor to align the fluid flow with the center of a port at the distal end of the housing. In other embodiments, the system includes a memory having instructions stored thereon, which, when executed by the processor, cause the processor to align the fluid flow with the center of the distal end of the housing. The system of interest is configured for sorting particles of a sample (e.g., a biological sample) in a fluid stream. In some embodiments, the system further includes a sample fluid delivery subsystem and a sheath fluid delivery subsystem, the sheath fluid delivery subsystem being in fluid communication with an inlet at the proximal end of the particle sorting module. In some cases, a waste bin is coupled to an outlet of the particle sorting module.

[0008] Aspects of the present disclosure also include methods for aligning a fluid stream with one or more ports at the distal end of a particle sorting module. The method according to certain embodiments includes: capturing an image through an opening in a particle sorting module, the opening being configured to visualize droplets of a fluid stream; calculating the pixel position of the boundary of each port at the distal end in the imaging opening along a horizontal axis; and aligning the fluid stream with the port based on the calculated pixel position of the boundary of each port in the imaging opening. In some embodiments, calculating the pixel position includes: calculating the total number of pixels passing through the imaging opening along the horizontal axis; and multiplying the total number of pixels by a predetermined ratio. In some cases, the method further includes calculating a predetermined ratio by dividing the number of pixels of each port boundary in the imaging opening by the total number of pixels passing through the imaging opening along the horizontal axis. In some embodiments, the method includes irradiating the fluid stream through the opening with a light source (e.g., a laser). The method according to certain instances also includes sorting particles in the fluid stream. In some embodiments, the method includes: irradiating a sample in an interrogation region of a particle sorting module with a light source, the sample having particles in the fluid stream; detecting light from the sample and applying sufficient deflection force to the particles in the fluid stream to separate the particles into one or more sample containers at the distal end of the particle sorting module. In certain embodiments, the sample is a biological sample and the method comprises sorting and collecting two or more different types of cells.

[0009] Also provided are kits having one or more subject particle sorting modules. Kits according to certain embodiments include a particle sorting module comprising an opening configured for visualizing droplets of a deflected fluid stream and one or more sample collection containers for coupling to the particle sorting module. Kits of interest may also include a droplet deflector. In some embodiments, the particle sorting module includes one or more of a sample inlet and a sheath fluid inlet, and the kit may include one or more connectors for coupling the particle sorting module to a sample fluid delivery system and a sheath fluid delivery system. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The present invention may be best understood from the following detailed description when read in conjunction with the accompanying drawings. Included in the drawings are the following figures:

[0011] Figure 1 Depicted are example calculations for determining the boundaries of a sample collection port at the distal end of a particle sorting module in accordance with certain embodiments.

[0012] Figure 2 Depicted is a particle sorting module having openings configured to visualize droplets of a deflected fluid stream, according to certain embodiments. DETAILED DESCRIPTION

[0013] Aspects of the present disclosure include a particle sorting module having an opening configured to visualize droplets of a deflected fluid stream. A particle sorting module according to certain embodiments includes a housing having a proximal end, a distal end, and a wall therebetween, the wall having an opening positioned in the wall, the opening being configured to align the fluid stream with one or more sample containers at the distal end. Systems and methods for aligning a fluid stream with one or more sample containers and sorting particles of a sample (e.g., a biological sample) are also provided. A kit having one or more particle sorting modules is also described, the particle sorting modules being suitable for coupling with a particle sorting system and being suitable for implementing the subject method.

[0014] Before describing the present invention in more detail, it should be understood that the present invention is not limited to the specific embodiments described, as these embodiments will of course vary. It should also be understood that the terminology used herein is only for the purpose of describing specific embodiments and is not intended to be limiting, as the scope of the present invention will only be limited by the appended claims.

[0015] Where a range of values ​​is provided, it is to be understood that, unless the context clearly dictates otherwise, each intermediate value of one tenth of the unit of the lower limit between the upper and lower limits of the range and any other stated or intermediate values ​​within the stated range are included within the present invention. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges and are also included within the present invention, subject to any specific exclusions in the stated ranges. Where the stated range includes one or two limitations, ranges not including one or both of these included limitations are also included within the present invention.

[0016] Certain ranges appear herein with numerical values ​​preceded by the term "about". The term "about" is used herein to provide literal support for the exact number preceding it, as well as numbers that are close to or approximately the number preceding the term. In determining whether a number is close to or approximately a specifically recited number, the close or approximate unrecited number may be a number that, in the context in which it appears, provides a substantial equivalence to the specifically recited number.

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative illustrative methods and materials are now described.

[0018] All publications and patents cited in this specification are incorporated herein by reference, as if each individual publication or patent was specifically and individually indicated to be incorporated by reference, and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with the cited publications. The citation of any publication is due to its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. In addition, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0019] It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the claims may be drafted to exclude any optional elements. Therefore, this statement is intended to serve as antecedent basis for using terminology such as "solely," "only," and the like when reciting claim elements or for using a "negative" limitation.

[0020] As will be clear to those skilled in the art upon reading this disclosure, each of the various embodiments described and illustrated herein has discrete components and features that can be easily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any method cited can be executed in the order of events cited or in any other order that is logically possible.

[0021] Although the apparatus and methods have been or will be described using functional interpretations for grammatical fluency, it should be clearly understood that, unless expressly stated in accordance with 35 U.S.C. §112, the claims should not be interpreted as necessarily being limited in any way to constructions of “means” or “step” limitations, but should be given the full range of meanings and equivalent meanings defined in the claims under the doctrine of judicial equivalence, and if the claims are expressly stated in accordance with 35 U.S.C. §112, they should be given all legal equivalent meanings in accordance with 35 U.S.C. §112.

[0022] As described above, the present invention provides a particle sorting module for sorting particle components of a sample, such as cells in a biological sample. In further describing an embodiment of the present invention, a particle sorting module having an opening is first described in more detail, wherein the opening is configured to visualize droplets of a deflected fluid stream (e.g., to align the deflected fluid stream with one or more sample collection containers). Next, a particle sorting system and method for separating particles in a sample are described. A kit having one or more particle sorting modules is also provided, wherein the particle sorting module is suitable for coupling with a particle sorting system and suitable for implementing the subject method.

[0023] Particle sorting module with openings for visualizing fluid flow

[0024] As described above, aspects of the present invention include a particle sorting module having an opening configured to visualize droplets of a deflected fluid stream. In an embodiment, the subject particle sorting module is configured to sort components of a sample, such as cells in a biological sample. The term "sorting" is used herein in its traditional sense to refer to components (e.g., cells, non-cellular particles, such as biomacromolecules) of a separated sample, and in some cases, as described below, the separated components are transported to one or more ports that are coupled to a sample collection container. For example, the subject particle sorting module can be configured to sort a sample having 2 or more components, such as 3 or more components, such as 4 or more components, such as 5 or more components, such as 10 or more components, such as 15 or more components, and includes sorting a sample having 25 or more components. One or more sample components can be separated from the sample and transported to a sample collection container, for example, 2 or more sample components, for example, 3 or more sample components, for example, 4 or more sample components, for example, 5 or more sample components, for example, 10 or more sample components, and including 15 or more sample components can be separated from the sample and transported to a sample collection container at a receiving position.

[0025] In an embodiment, the particle sorting module includes an opening (e.g., as described in more detail below) for visualizing droplets deflected by a deflection force applied from one or more droplet deflectors. In some embodiments, the particle sorting module is configured to generate an analyzed droplet stream and deflect each analyzed droplet from the analyzed droplet stream to a deflected droplet receiving position. As used herein, the term "deflected droplet receiving position" refers to a position downstream of a droplet deflector at which sorted droplets containing particles of interest (e.g., cells) can be collected after they are deflected by a droplet deflector plate. In an embodiment, droplets in a fluid stream deviate from their normal trajectory along a longitudinal axis of the fluid stream. In some cases, droplets in a fluid stream deviate from their normal trajectory at a fluid stream demarcation point. "Demarcation point" refers to a point in a fluid stream where a continuous fluid stream begins to form discrete droplets. In other cases, a droplet in the fluid stream deviates from its normal trajectory at a location downstream from the demarcation point, for example, deviates from the demarcation point by 0.001 mm or more, for example, 0.005 mm or more, for example, 0.01 mm or more, for example, 0.05 mm or more, for example, 0.1 mm or more, for example, 0.5 mm or more, for example, 1 mm or more, for example, 2 mm or more, for example, 5 mm or more, for example, 10 mm or more, for example, 15 mm or more, for example, 20 mm or more, for example, 25 mm or more, for example, 30 mm or more, for example, 35 mm or more, and includes a droplet deviating from its normal trajectory downstream of the fluid stream demarcation point by a droplet deflector by 50 mm or more.

[0026] In embodiments, a droplet is deflected from its normal trajectory by a droplet deflector a distance of 0.001 mm or more, as measured radially in a plane orthogonal to the longitudinal axis of the fluid flow, such as 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 2 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 20 mm or more, such as 25 mm or more, such as 30 mm or more, such as 35 mm or more, and including 50 mm or more. For example, droplets in a fluid stream may deviate a distance from 0.001 mm to 100 mm, for example from 0.005 mm to 95 mm, for example from 0.001 mm to 90 mm, for example from 0.05 mm to 85 mm, for example from 0.01 mm to 80 mm, for example from 0.05 mm to 75 mm, for example from 0.1 mm to 70 mm, for example from 0.5 mm to 65 mm, for example from 1 mm to 60 mm, for example from 5 mm to 55 mm, and including from 10 mm to 50 mm. Thus, the angle at which droplets in the fluid stream deviate from a trajectory along the longitudinal axis of the fluid stream is from 0.01° to 90°, for example from 0.05° to 85°, for example from 0.1° to 80°, for example from 0.5° to 75°, for example from 10° to 70°, for example from 15° to 65°, for example from 20° to 60°, for example from 25° to 55°, and including from 30° to 50°.

[0027] The particle sorting module of interest may include one or more sample collection ports at a droplet receiving position (e.g., at the distal end of the particle sorting module). Each sample collection port includes an opening (i.e., an empty space through which droplets pass) for receiving droplets from a fluid stream. In an embodiment, the opening of each sample collection port at the distal end of the particle sorting module described herein may be independently 0.001 mm or more, such as 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 2 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 20 mm or more, such as 25 mm or more, such as 30 mm or more, such as 35 mm or more, and includes a distance of 50 mm or more from the longitudinal axis of the fluid stream. For example, the opening of each sample collection port at the distal end of the particle sorting module can range from 0.001 mm to 100 mm, such as from 0.01 mm to 90 mm, such as from 0.1 mm to 80 mm, such as from 0.5 mm to 70 mm, such as from 1 mm to 60 mm, and including from 5 mm to 50 mm. The opening of each sample collection port at the distal end of the particle sorting module can each independently form a different angle relative to the longitudinal axis of the fluid flow, such as 0.01° to 90°, such as from 0.05° to 85°, such as from 0.1° to 80°, such as from 0.5° to 75°, such as from 10° to 70°, such as from 15° to 65°, such as from 20° to 60°, such as from 25° to 55°, and including from 30° to 50°. In an embodiment, where the distal end of the particle sorting module includes more than one sample collection port, the space between the openings of each sample collection port can independently be 0.001 mm or more, such as 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 2 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 20 mm or more, such as 30 mm or more, such as 35 mm or more, and including 50 mm or more from the longitudinal axis of the fluid flow. For example, the space between the openings of each sample collection port can independently vary from 0.001 mm to 100 mm, such as from 0.01 mm to 90 mm, such as from 0.1 mm to 80 mm, such as from 0.5 mm to 70 mm, such as from 1 mm to 60 mm, and including from 5 mm to 50 mm.

[0028] In some cases, the opening of each sample collection port may include a cover, such as a valve, that can be reversibly closed as needed. For example, the opening of the sample collection port may include a valve that is configured to remain closed until fluid flow is detected. In other cases, the opening of the sample collection port includes a valve that is configured to remain closed until a sample collection container is coupled to the sample collection port. In other cases, the opening of the sample collection port includes a valve that is configured to remain closed after a predetermined time period (e.g., to collect droplets only from a specific portion of the sample). In other cases, the opening of the sample collection port includes a valve that is configured to close after detecting that a sample collection container coupled to the sample collection port has collected a predetermined volume. In some cases, the opening of the sample collection port is configured to close in response to a command input by a user.

[0029] The opening of the sample collection port at the distal end of the particle sorting module of interest can be any suitable shape, wherein the shape of the opening of the sample collection port of interest includes: a straight shape, such as a square, rectangle, trapezoid, triangle, hexagon, etc.; a curved shape, such as a circle, an ellipse; and an irregular shape, for example, a parabola bottom coupled to a plane top. In some embodiments, the opening of the sample collection port has a circular shape. In other embodiments, the opening of the sample collection port has an elliptical shape. In other embodiments, the opening of the sample collection port has a square or rectangular shape.

[0030] The size of the opening of each sample collection port can vary, with the size of each opening independently ranging from 0.1 cm to 5 cm, such as from 0.25 cm to 4.5 cm, such as from 0.5 cm to 4 cm, such as from 0.75 cm to 3.5 cm, and including from 1 cm to 3 cm. In the case where the opening of the sample collection port is circular, the diameter of the opening can range from 0.1 cm to 5 cm, such as from 0.25 cm to 4.5 cm, such as from 0.5 cm to 4 cm, such as from 0.75 cm to 3.5 cm, and including from 1 cm to 3 cm in diameter.

[0031] The particle sorting module of interest may include one or more sample collection ports at the distal end of the housing, such as 2 or more, such as 3 or more, such as 4 or more, such as 5 or more, such as 6 or more, such as 7 or more, such as 8 or more, such as 9 or more, and including 10 or more sample collection ports. In the case where the sample collection port is coupled to a sample collection container (e.g., a waste collection container), the particle sorting module may include one or more sample collection containers coupled to the particle sorting module housing, such as 2 or more, such as 3 or more, such as 4 or more, such as 5 or more, such as 6 or more, such as 7 or more, such as 8 or more, such as 9 or more, and including 10 or more sample collection containers.

[0032] The sample collection container can be coupled to the sample collection port at the distal end of the particle sorting module by any convenient attachment scheme. In some embodiments, the sample collection container is mechanically coupled to the sample collection port, for example, by a luer-lok connection, by aseptic tube welding or by threading to the sample collection port. In other embodiments, the sample collection container is fixed to the sample collection port by a permanent or non-permanent adhesive. In other embodiments, the sample collection container is co-molded with the sample collection port. In other embodiments, the sample collection container is integrated with the particle sorting module housing so that the sample collection container and the particle sorting module housing form a single unit. In other cases, the sample collection container can be fluidically coupled to the sample collection port, for example via a tube configured to transport sorted droplets, wherein these embodiments can provide aseptic recovery of sorted droplets, for example, by clamping and cutting off a fluid transport structure (e.g., a tube). Suitable sample collection containers for collecting droplets from a fluid stream may include, but are not limited to, test tubes, conical tubes, multi-chamber containers (e.g., microtiter plates (e.g., 96-well plates)), centrifuge tubes, culture tubes, microtubes, caps, test tubes, bottles, linear polymer containers, and bags.

[0033] Transferring the droplets of interest to a sample collection container (eg, through a sample collection port) can be accomplished by a droplet deflector via electrostatic charging of the droplets and deflecting the charged droplets from the fluid stream by applying an electrostatic field. The voltage applied to the metal plate of the droplet deflector in the subject sorting module can be 10mV or greater, for example 25mV or greater, for example 50mV or greater, for example 75mV or greater, for example 100mV or greater, for example 250mV or greater, for example 500mV or greater, for example 750mV or greater, for example 1V or greater, for example 2.5V or greater, for example 5V or greater, for example 10V or greater, for example 25V or greater, for example 50V or greater, and including 100V or greater, for example 500V or greater, for example 1000V or greater, for example 5000V or greater, for example 10000V or greater, for example 15000V or greater, for example 25000V or greater, for example 50000V or greater, and including 100000V or greater. In certain embodiments, the voltage applied to each set of parallel metal plates is from 0.5 kV to 15 kV, such as from 1 kV to 15 kV, such as from 1.5 kV to 12.5 kV, and including from 2 kV to 10 kV.

[0034] Particles in the fluid stream may be deflected by any convenient deflector plate arrangement, including but not limited to those described in U.S. Patent Nos. 9,952,076, 9,933,341, 9,726,527, 9,453,789, 9,200,334, 9,097,640, 9,095,494, 9,092,034, 8,975,595, 8,753,573, 8,233,146, 8,140 ,300, 7,544,326, 7,201,875, 7,129,505, 6,821,740, 6,813,017, 6,809,804, 6,372,506, 5,700,692, 5,643,796, 5,627,040, 5,620,842, 5,602,039, the entire disclosures of which are incorporated herein by reference. In certain embodiments, the deflector plate comprises a charge plate for sorting cells in a fluid stream, such as used in a flow cytometer system, such as a BD Biosciences FACSCanto TM II flow cytometer, BD Accuri TM Flow cytometer, BD Biosciences FACSCelesta TM Flow cytometer, BD Biosciences FACSLyric TM Flow cytometer, BD Biosciences FACSVerse TMFlow cytometer, BD Biosciences FACSymphony TM Flow cytometer, BD Biosciences LSRFortessa TM Flow cytometer, BD Biosciences LSR Fortess TM X-20 Flow Cytometer and BD Biosciences FACSCalibur TM Cell sorter, BD Biosciences FACSCount TM Cell sorter, BD Biosciences FACSLyric TM Cell Sorter and BD Biosciences Via TM Cell sorter, BD Biosciences Influx TM Cell sorter, BD Biosciences Jazz TM Cell sorter, BD Biosciences Aria TM Cell Sorter and BD Biosciences FACSMelody TM Cell sorting machine, etc.

[0035] The opening in the particle sorting module as described herein is configured to visualize deflected droplets and, in some cases, to visualize a sample collection container at a droplet receiving position. As described above, in some embodiments, the particle sorting module includes a housing having a proximal end, a distal end, and a wall therebetween, the wall having an opening positioned in the wall, the opening being used to visualize deflected droplets. The opening can be any suitable shape, wherein the shapes used to visualize the opening of interest include, but are not limited to: a straight shape, such as a square, rectangle, trapezoid, triangle, hexagon, etc.; a curved shape, such as a circle, an ellipse; and an irregular shape, such as a parabola bottom coupled to a plane top. In some embodiments, the opening in the wall of the particle sorting module housing has a circular shape. In other embodiments, the opening in the wall of the particle sorting module housing has an elliptical shape. In other embodiments, the opening in the wall of the particle sorting module housing has a square or rectangular shape.

[0036] In some embodiments, the size of the opening in the wall of the particle sorting module housing can vary, with a width ranging from 0.5cm to 10cm, for example from 1cm to 9.5cm, for example from 1.5cm to 9cm, for example from 2cm to 8.5cm, for example from 2.5cm to 8cm, for example from 3cm to 7.5cm, for example from 3.5cm to 7cm, for example from 4cm to 6.5cm, and including from 4cm to 6cm, for example 5cm, and a length ranging from 1cm to 20cm, for example from 1.5cm to 19.5cm, for example from 2cm to 19cm, for example from 2.5cm to 18.5cm, for example from 3cm to 18cm, for example 3.5cm to 17.5cm, for example from 4cm to 17cm, for example from 4.5cm to 16.5cm, and including from 5cm to 15cm, for example 10cm. In some embodiments, the opening in the wall of the particle sorting module housing is circular and the diameter of the opening ranges from 0.5 cm to 10 cm, such as from 1 cm to 9.5 cm, such as from 1.5 cm to 9 cm, such as from 2 cm to 8.5 cm, such as from 2.5 cm to 8 cm, such as from 3 cm to 7.5 cm, such as from 3.5 cm to 7 cm, such as from 4 cm to 6.5 cm, and including from 4 cm to 6 cm, such as 5 cm. As described in more detail below, in some cases, an image is captured through the opening in the particle sorting module and the number of pixels passing through the horizontal axis of the opening is counted. Depending on the size of the opening and the size of each pixel, the opening in the wall of the particle sorting module housing can vary and in some cases is configured to provide a range from 1x10 1 Pixels up to 1x10 6 Pixel images, for example from 5x10 1 Pixels up to 5x10 5 Pixels, for example from 1x10 2 Pixels up to 1x10 5 Pixels, for example from 5x10 2 Pixels up to 5x10 4 Pixels, including from 1x10 3 Pixels up to 1x10 4 Pixel.

[0037] The opening in the wall of the particle sorting module housing is positioned between the droplet deflector and the distal end of the housing. Depending on the length of the particle sorting module, in some embodiments, the upper boundary of the opening is located at a position from 0.5 cm to 25 cm from the bottom of the droplet deflector, such as from 1 cm to 24.5 cm, such as from 1.5 cm to 24 cm, such as from 2 cm to 23.5 cm, such as from 2.5 cm to 23 cm, such as from 3 cm to 22.5 cm, such as from 3.5 cm to 22 cm, such as from 4 cm to 21.5 cm, such as from 4.5 cm to 21 cm, and the upper boundary of the opening in the wall of the particle sorting module housing is located at a position from 5 cm to 20 cm from the bottom of the droplet deflector. In some embodiments, the lower boundary (i.e., bottom) of the opening in the wall of the particle sorting module housing is located at a position from 0.5 cm to 10 cm from the far end of the housing, for example from 1 cm to 9.5 cm, for example from 1.5 cm to 9 cm, for example from 2 cm to 8.5 cm, for example from 2.5 cm to 8 cm, for example from 3 cm to 7.5 cm, for example from 3.5 cm to 7 cm, for example from 4 cm to 6.5 cm, and the lower boundary (i.e., bottom) of the opening in the wall of the particle sorting module housing is positioned from 4 cm to 6 cm, for example 5 cm.

[0038] In some embodiments, the opening in the wall of the particle sorting module housing includes one or more optical adjustment components. The so-called "optical adjustment" refers to: changing the light propagating through the opening or the light collected from the irradiated fluid flow through the opening as needed. The optical adjustment component can be any convenient device or structure that provides the required change of light, and can include but is not limited to optical windows (e.g., transparent windows) lenses, mirrors, pinholes, slits, gratings, light refractors and any combination thereof. The opening in the wall of the particle sorting module housing can include one or more optical adjustment components at the sample interrogation area as needed, such as two or more, such as three or more, such as four or more, and including five or more optical adjustment components.

[0039] In some cases, the opening in the wall of the particle sorting module housing includes a transparent optical window. By "optically transparent" is meant that the optical window does not restrict the propagation of at least one or more wavelengths of light therein. In some embodiments, the optical window is optically transparent to one or more of ultraviolet light, visible light, and near-infrared light. In one example, the optical window is transparent to ultraviolet light. In another example, the optical window is transparent to visible light. In yet another example, the optical window is transparent to near-infrared light. In yet another example, the optical window is transparent to ultraviolet light and visible light. In yet another example, the optical window is transparent to visible light and near-infrared light. In yet another example, the optical window is transparent to ultraviolet light, visible light, and near-infrared light. Depending on the desired light propagation characteristics, the transparent optical window can be formed of any suitable material, such as quartz, glass, silicon dioxide, titanium dioxide, zinc oxide, silicon nitride, carbon black, aluminum oxide, or polymers, including but not limited to optically transparent polymers, such as optically transparent polymers such as acrylic acid, acrylic acid / styrene, cycloolefin polymers, polycarbonate, polyester, and polystyrene. The optically transparent region can extend across 5% or more of the optical window, such as across 10% or more, such as across 25% or more, such as across 50% or more, such as across 75% or more, such as across 90% or more, such as across 95% or more, and including across 99% or more of the optical window. In some embodiments, the entire optical window (i.e., 100%) is optically transparent. For example, the optically transparent region can extend across from 5% to 100% of the optical window, such as from 10% to 90%, such as from 20% to 80%, such as from 30% to 70%, and including from 40% to 60%.

[0040] In some embodiments, the optical window includes one or more reference identifiers. In some cases, the reference identifier may include a mark associated with the boundary of the deflected fluid flow. For example, these marks depict the deflection path of the fluid flow. For example, these marks can be associated with the charge of the droplet deflector plate or the voltage applied to the droplet deflector plate. In other cases, the mark is associated with the boundary of one or more ports at the distal end of the housing. For example, the optical window may include a reference identifier that depicts the boundary of one or more ports coupled to the sample collection container. In some cases, the optical window includes a reference identifier for aligning the fluid flow with the one or more ports. For example, the optical window may include a reference identifier that indicates the position of the waste collection port at the distal end of the particle sorting module housing. In some embodiments, the optical window includes a reference identifier indicating the center of the distal end of the housing. For example, the optical window may include a reference identifier for aligning the fluid flow with the center of the distal end of the particle sorting module housing.

[0041] In some embodiments, the opening in the particle sorting module housing includes an optical adjustment component for adjusting the light irradiated by a light source (e.g., a laser) onto the deflected fluid flow. For example, the optical adjustment can be to increase the size of the light, the focus of the light, or to collimate the light. In some cases, the optical adjustment is an amplification scheme to increase the size of the light (e.g., a light spot), such as increasing the size by 5% or more, such as 10% or more, such as 25% or more, such as 50% or more, and including increasing the size by 75% or more. In other embodiments, the optical adjustment includes focusing the collected light to reduce the size of the light, such as by 5% or more, such as by 10% or more, such as by 25% or more, such as by 50% or more, and including reducing the size of the light spot by 75% or more. In certain embodiments, the optical adjustment includes collimation of the light. The term "collimation" in its conventional sense refers to reducing divergence by optically adjusting the collinearity of light propagation or by light from a common propagation axis. In some cases, collimation includes reducing the spatial cross-section of the light beam (e.g., reducing the beam profile of the laser).

[0042] In some embodiments, the opening in the particle sorting module housing includes: a focusing lens having a magnification from 0.1 to 0.95, such as a magnification from 0.2 to 0.9, such as a magnification from 0.3 to 0.85, such as a magnification from 0.35 to 0.8, such as a magnification from 0.5 to 0.75, and including a magnification from 0.55 to 0.7, such as a magnification of 0.6. For example, in some cases, the focusing lens is a bichromatic de-magnifying lens with a magnification of approximately 0.6. The focal length of the focusing lens can vary, ranging from 5mm to 20mm, such as from 6mm to 19mm, such as from 7mm to 18mm, such as from 8mm to 17mm, such as from 9mm to 16, and including a focal length ranging from 10mm to 15mm. In some embodiments, the focusing lens has a focal length of approximately 13mm.

[0043] In other embodiments, the optical adjustment component is a collimator. The collimator can be any convenient collimation scheme, such as one or more mirrors or curved lenses or a combination thereof. For example, the collimator is a single collimating lens in some cases. In other cases, the collimator is a collimating mirror. In other cases, the collimator includes two lenses. In other cases, the collimator includes a mirror and a lens. In the case where the collimator includes one or more lenses, the focal length of the collimating lens can vary, ranging from 5mm to 40mm, such as from 6mm to 37.5mm, such as from 7mm to 35mm, such as from 8mm to 32.5mm, such as from 9mm to 30mm, such as from 10mm to 27.5mm, such as from 12.5mm to 25mm, and including focal lengths ranging from 15mm to 20mm.

[0044] In some embodiments, the particle sorting module also includes a fluid pool nozzle having a nozzle orifice, the nozzle orifice being configured to cause a fluid flow to flow through the fluid pool nozzle and one or more of the above-mentioned subject droplet deflectors, the droplet deflectors being configured to apply a deflection force to the fluid flow flowing therebetween. The subject particle sorting module includes a fluid pool nozzle having an orifice for propagating a fluid sample to a sample interrogation area, wherein, in some embodiments, the fluid pool nozzle includes: a proximal cylindrical portion defining a longitudinal axis; and a distal frustoconical portion terminating in a plane having a nozzle orifice transverse to the longitudinal axis. The length of the proximal cylindrical portion (as measured along the longitudinal axis) can vary, ranging from 1 mm to 15 mm, for example, from 1.5 mm to 12.5 mm, for example, from 2 mm to 10 mm, for example, from 3 mm to 9 mm, and including from 4 mm to 8 mm. The length of the distal frustoconical portion (as measured along the longitudinal axis) can also vary, ranging from 1 mm to 10 mm, such as from 2 mm to 9 mm, such as from 3 mm to 8 mm, and including from 4 mm to 7 mm. In some embodiments, the diameter of the fluid pool nozzle chamber can vary, ranging from 1 mm to 10 mm, such as from 2 mm to 9 mm, such as from 3 mm to 8 mm, and including from 4 mm to 7 mm.

[0045] In some cases, the nozzle chamber does not include a cylindrical portion, and the entire fluid pool nozzle chamber is frustoconical. In these embodiments, the length of the frustoconical nozzle chamber (as measured along the longitudinal axis transverse to the nozzle orifice) can range from 1mm to 15mm, such as from 1.5mm to 12.5mm, such as from 2mm to 10mm, such as from 3mm to 9mm, and includes from 4mm to 8mm. The diameter of the proximal portion of the frustoconical nozzle chamber can range from 1mm to 10mm, such as from 2mm to 9mm, such as from 3mm to 8mm, and includes from 4mm to 7mm.

[0046] In an embodiment, the sample fluid stream is emitted from an orifice at the distal end of the fluid pool nozzle. Depending on the desired characteristics of the fluid stream, the fluid pool nozzle orifice can be any suitable shape, where the cross-sectional shapes of interest include, but are not limited to: straight cross-sectional shapes, such as square, rectangular, trapezoidal, triangular, hexagonal, etc.; curved cross-sectional shapes, such as circular, elliptical; and irregular shapes, such as a parabolic bottom coupled to a flat top. In some embodiments, the fluid pool nozzle of interest has a circular orifice. In some embodiments, the size of the nozzle orifice can vary, ranging from 1 μm to 20000 μm, such as from 2 μm to 17500 μm, such as from 5 μm to 15000 μm, such as from 10 μm to 12500 μm, such as from 15 μm to 10000 μm, such as from 25 μm to 7500 μm, such as from 50 μm to 5000 μm, such as from 75 μm to 1000 μm, such as from 100 μm to 750 μm, and including from 150 μm to 500 μm. In some embodiments, the nozzle orifice is 100 μm.

[0047] In some embodiments, the fluid pool nozzle includes a sample injection port configured to provide a sample to the fluid pool nozzle. In an embodiment, the sample injection system is configured to provide a suitable sample fluid to the fluid pool nozzle chamber. Depending on the desired characteristics of the fluid flow, the sample rate delivered to the fluid pool nozzle chamber through the sample injection port can be 1 μL / sec or greater, such as 2 μL / sec or greater, such as 3 μL / sec or greater, such as 5 μL / sec or greater, such as 10 μL / sec or greater, such as 15 μL / sec or greater, such as 25 μL / sec or greater, such as 50 μL / sec or greater, such as 100 μL / sec or greater, such as 150 μL / sec or greater, such as 200 μL / sec or greater, such as 250 μL / sec or greater, such as 300 μL / sec or greater, such as 350 μL / sec or greater, such as 400 μL / sec or greater, such as 450 μL / sec or greater, and including 500 μL / sec or greater. For example, the sample flow rate can range from 1 μL / sec to about 500 μL / sec, for example from 2 μL / sec to about 450 μL / sec, for example from 3 μL / sec to about 400 μL / sec, for example from 4 μL / sec to about 350 μL / sec, for example from 5 μL / sec to about 300 μL / sec, for example from 6 μL / sec to about 250 μL / sec, for example from 7 μL / sec to about 200 μL / sec, for example from 8 μL / sec to about 150 μL / sec, for example from 9 μL / sec to about 125 μL / sec, and including from 10 μL / sec to about 100 μL / sec.

[0048] The sample injection port can be an orifice located in the wall of the nozzle chamber, or can be a conduit located at the proximal end of the nozzle chamber. In the case where the sample injection port is an orifice located in the wall of the nozzle chamber, the sample injection port orifice can be any suitable shape, wherein the cross-sectional shape of interest includes, but is not limited to: a straight cross-sectional shape, such as a square, rectangle, trapezoid, triangle, hexagon, etc.; a curved cross-sectional shape, such as a circle, an ellipse, etc.; and an irregular shape, such as a parabolic bottom coupled to a plane top. In certain embodiments, the sample injection port has a circular orifice. In certain cases, the size of the sample injection port orifice can vary according to the shape, and its opening range is from 0.1mm to 5.0mm, such as 0.2mm to 3.0mm, such as 0.5mm to 2.5mm, such as from 0.75mm to 2.25mm, such as from 1mm to 2mm, and includes from 1.25mm to 1.75mm, such as 1.5mm.

[0049] In some cases, the sample injection port is a conduit located at the proximal end of the fluid pool nozzle chamber. For example, the sample injection port can be a conduit positioned so that the orifice of the sample injection port is aligned with the fluid pool nozzle orifice. In the case where the sample injection port is a conduit positioned to align with the fluid pool nozzle orifice, the cross-sectional shape of the sample injection tube can be any suitable shape, wherein the cross-sectional shape of interest includes but is not limited to: a straight cross-sectional shape, such as a square, rectangle, trapezoid, triangle, hexagon, etc.; a curved cross-sectional shape, such as a circle, an ellipse; and an irregular shape, such as a parabola bottom coupled to a plane top. In some cases, the orifice of the conduit can vary according to the shape, and its opening range is from 0.1mm to 5.0mm, such as 0.2mm to 3.0mm, such as 0.5mm to 2.5mm, such as from 0.75mm to 2.25mm, such as from 1mm to 2mm, and includes from 1.25mm to 1.75mm, such as 1.5mm. The shape of the tip of the sample injection port can be the same or different from the cross-sectional shape of the sample injection tube. For example, the orifice of the sample injection port may include a bevel tip having a bevel angle ranging from 1° to 10°, such as from 2° to 9°, such as from 3° to 8°, such as from 4° to 7°, and including a bevel angle of 5°.

[0050] In some embodiments, the fluid pool nozzle also includes a sheath liquid injection port configured to provide sheath liquid to the fluid pool nozzle. In an embodiment, the sheath liquid injection system is configured to provide a fluid of sheath liquid to the fluid pool nozzle chamber, such as to produce a laminar fluid flow of sheath liquid surrounding the sample fluid flow together with the sample. Depending on the desired characteristics of the fluid flow, the rate of sheath liquid delivered to the fluid pool nozzle chamber through the tube can be 25 μL / sec or greater, such as 50 μL / sec or greater, such as 75 μL / sec or greater, such as 100 μL / sec or greater, such as 250 μL / sec or greater, such as 500 μL / sec or greater, such as 750 μL / sec or greater, such as 1000 μL / sec or greater, and including 2500 μL / sec or greater. For example, the sheath fluid flow rate can range from 1 μL / sec to about 500 μL / sec, for example from 2 μL / sec to about 450 μL / sec, for example from 3 μL / sec to about 400 μL / sec, for example from 4 μL / sec to about 350 μL / sec, for example from 5 μL / sec to about 300 μL / sec, for example from 6 μL / sec to about 250 μL / sec, for example from 7 μL / sec to about 200 μL / sec, for example from 8 μL / sec to about 150 μL / sec, for example from 9 μL / sec to about 125 μL / sec, and including from 10 μL / sec to about 100 μL / sec.

[0051] In some embodiments, the sheath injection port is an orifice located in the wall of the nozzle chamber. The sheath injection port orifice can be any suitable shape, where the cross-sectional shapes of interest include, but are not limited to: straight cross-sectional shapes, such as squares, rectangles, trapezoids, triangles, hexagons, etc.; curved cross-sectional shapes, such as circles, ellipses; and irregular shapes, such as a parabolic bottom coupled to a flat top. In some cases, the size of the sample injection port orifice can vary depending on the shape, with an opening ranging from 0.1 mm to 5.0 mm, such as 0.2 mm to 3.0 mm, such as 0.5 mm to 2.5 mm, such as from 0.75 mm to 2.25 mm, such as from 1 mm to 2 mm, and including from 1.25 mm to 1.75 mm, such as 1.5 mm.

[0052] The particle sorting module also includes a sample interrogation area in fluid communication with the fluid pool nozzle orifice. As described in more detail below, a sample fluid stream is emitted from an orifice at the distal end of the fluid pool nozzle, and particles in the fluid stream can be irradiated with a light source at the sample interrogation area of ​​the particle sorting module. The size of the interrogation area of ​​the particle sorting module can vary according to the characteristics of the fluid nozzle, such as the size of the nozzle orifice and the size of the sample injection port. In an embodiment, the width of the interrogation area can be 0.01 mm or greater, such as 0.05 mm or greater, such as 0.1 mm or greater, such as 0.5 mm or greater, such as 1 mm or greater, such as 2 mm or greater, such as 3 mm or greater, such as 5 mm or greater, and including 10 mm or greater. The length of the interrogation zone may also vary, in some cases ranging from 0.01 mm or greater, such as 0.1 mm or greater, such as 0.5 mm or greater, such as 1 mm or greater, such as 1.5 mm or greater, such as 2 mm or greater, such as 3 mm or greater, such as 5 mm or greater, such as 10 mm or greater, such as 15 mm or greater, such as 20 mm or greater, such as 25 mm or greater, and including particle sorting modules of 50 mm or greater.

[0053] The interrogation region on the particle sorting module can be configured to facilitate irradiation of a planar cross-section of the emitted fluid stream, or can be configured to facilitate irradiation of a predetermined length of a diffuse field (e.g., with a diffuse laser or lamp). In some embodiments, the interrogation region on the particle sorting module includes a transparent window that facilitates irradiation of a predetermined length of the emitted fluid stream, such as 1 mm or greater, such as 2 mm or greater, such as 3 mm or greater, such as 4 mm or greater, such as 5 mm or greater, and including 10 mm or greater. Depending on the light source used to irradiate the emitted fluid stream (described below), the interrogation region of the particle sorting module can be configured to pass light in the range of from 100 nm to 1500 nm, such as from 150 nm to 1400 nm, such as from 200 nm to 1300 nm, such as from 250 nm to 1200 nm, such as from 300 nm to 1100 nm, such as from 350 nm to 1000 nm, such as from 400 nm to 900 nm, and including from 500 nm to 800 nm.Thus, the particle sorting module at the interrogation region may be formed of any transparent material that passes the desired wavelength range, including but not limited to: optical glass, borosilicate glass, Pyrex glass, UV quartz, infrared quartz, sapphire, and plastics such as polycarbonate, polyvinyl chloride (PVC), polyurethane, polyether, polyamide, polyimide, or copolymers of these thermoplastics, such as PETG (ethylene glycol-modified polyethylene terephthalate), and other polymeric plastic materials including polyesters, where the polyesters of interest may include but are not limited to: poly(alkyl terephthalates), such as poly(ethylene terephthalate), (PET), bottle-grade PET (a copolymer based on monoethylene glycol, terephthalic acid and other comonomers such as isophthalic acid, cyclohexene dimethanol, etc.), poly(butylene terephthalate) (PBT) and poly(hexamethyl terephthalate); poly(alkyl adipates), such as poly(vinyl adipate), poly(1,4-butylene adipate) and poly(hexamethylene adipate); poly(alkyl suberates), such as poly(vinyl suberate); poly(alkyl sebacates), such as poly(vinyl sebacate); poly(ε-caprolactone) and poly(β-propiolactone); poly(isophthalic acid) alkyl esters), such as poly(isophthalate); poly(alkyl 2,6-naphthalene dicarboxylates), such as poly(vinyl 2,6-naphthalene dicarboxylate); poly(alkyl sulfonyl-4,4′-dibenzoate), such as poly(vinyl sulfonyl-4,4′-dibenzoate); poly(alkyl p-phenylene dicarboxylates), such as poly(vinyl p-phenylene dicarboxylate); poly(trans-1,4-cyclohexane dialkyl dicarboxylates), such as poly(trans-1,4-cyclohexane dialkyl dicarboxylate); poly(alkyl 1,4-cyclohexane-dimethyl dicarboxylates), such as poly(vinyl 1,4-cyclohexane-dimethyl dicarboxylate) ; poly(alkyl [2.2.2]-bicyclooctane-1,4-dimethyldicarboxylates), such as poly(vinyl [2.2.2]-bicyclooctane-1,4-dimethyldicarboxylate); lactic acid polymers and copolymers, such as (S)-polylactic acid, (R,S)-polylactic acid, poly(tetramethyl glycolide) and poly(lactide-co-glycolide); and polycarbonates of bisphenol A, 3,3′-dimethylbisphenol A, 3,3′,5,5′-tetrachlorobisphenol A, 3,3′,5,5′-tetramethylbisphenol A; polyamides, such as poly(terephthalamide); polyesters, such as polyethylene terephthalate, such as Mylar. TMPolyethylene terephthalate; etc. In some embodiments, the particle sorting module of interest includes a test tube located in the sample interrogation area. In an embodiment, the test tube can pass light ranging from 100nm to 1500nm, such as from 150nm to 1400nm, such as from 200nm to 1300nm, such as from 250nm to 1200nm, such as from 300nm to 1100nm, such as from 350nm to 1000nm, such as from 400nm to 900nm, and including from 500nm to 800nm.

[0054] In some embodiments, the particle sorting module has a closed housing. The term "closed" means that each component of the particle sorting module is completely contained within the housing, and the components are sealed or isolated from the surrounding environment. In other words, the components within the closed housing are not exposed to the external environment or are not in contact with the external environment. In some embodiments, the components contained within the housing are isolated from the gas environment of the surrounding environment (i.e., not exposed to the gas outside the housing). In other embodiments, the components contained within the housing are isolated from the fluid environment of the surrounding environment (i.e., not exposed to any fluid present outside the housing). In yet other embodiments, the components contained within the housing are sterile and isolated from live bacteria or other microorganisms present in the surrounding environment (i.e., sterile).

[0055] The size of the particle sorting module housing can vary, with a length ranging from 10 cm to 100 cm, such as from 15 cm to 95 cm, such as from 20 cm to 90 cm, such as from 25 cm to 85 cm, such as from 30 cm to 80 cm, such as from 35 cm to 75 cm, and including from 40 cm to 60 cm. The width of the particle sorting module housing can range from 1 cm to 25 cm, such as from 2 cm to 20 cm, such as from 3 cm to 15 cm, and including from 5 cm to 10 cm.

[0056] The housing can be formed of any suitable material compatible with the fluid sample (e.g., a biological sample), including metal, glass (e.g., Pyrex glass, borosilicate glass), ceramic, or plastic. In some embodiments, the particle sorting module housing is formed of plastic, such as a hard plastic, a polymer, or a thermoplastic material. For example, suitable plastics may include, but are not limited to, polycarbonate, polyvinyl chloride (PVC), polyurethane, polyether, polyamide, polyimide, or copolymers of these thermoplastics, such as PETG (ethylene glycol-modified polyethylene terephthalate), and other polymeric plastic materials. In certain embodiments, the particle sorting module housing is formed of polyester, where the polyesters of interest may include, but are not limited to: poly(alkyl terephthalates), such as poly(ethylene terephthalate) (PET), bottle-grade PET (a copolymer based on monoethylene glycol, terephthalic acid and other comonomers (e.g., isophthalic acid, cyclohexene dimethanol, etc.)), poly(butylene terephthalate) (PBT), and poly(hexamethyl terephthalate); poly(alkyl adipates), such as poly(hexamethyl adipate); esters), poly(1,4-butylene adipate) and poly(hexamethylene adipate); poly(alkyl suberates), such as poly(vinyl suberate); poly(alkyl sebacates), such as poly(vinyl sebacate); poly(ε-caprolactone) and poly(β-propiolactone); poly(alkyl isophthalates), such as poly(vinyl isophthalate); poly(alkyl 2,6-naphthalene dicarboxylates), such as poly(vinyl 2,6-naphthalene dicarboxylate); poly(alkyl sulfonyl-4,4′-dibenzoate), such as Poly(sulfonyl-4,4′-dibenzoic acid vinyl ester); poly(p-phenylene alkylene dicarboxylates), such as poly(p-phenylene dicarboxylic acid vinyl ester); poly(trans-1,4-cyclohexane dialkyl dicarboxylates), such as poly(trans-1,4-cyclohexane dialkyl dicarboxylates); poly(1,4-cyclohexane-dimethyl dicarboxylates), such as poly(1,4-cyclohexane-dimethyl dicarboxylates); poly([2.2.2]-bicyclooctane-1,4-dimethyl dicarboxylates), such as poly( [2.2.2]-bicyclooctane-1,4-dimethyldicarboxylic acid vinyl ester); lactic acid polymers and copolymers, such as (S)-polylactic acid, (R,S)-polylactic acid, poly(tetramethyl glycolide) and poly(lactide-co-glycolide); and polycarbonates of bisphenol A, 3,3′-dimethylbisphenol A, 3,3′,5,5′-tetrachlorobisphenol A, 3,3′,5,5′-tetramethylbisphenol A; polyamides, such as poly(terephthalamide); polyesters, such as polyethylene terephthalate, such as Mylar TM Polyethylene terephthalate; etc.

[0057] In some embodiments, one or more outer walls of the particle sorting module housing have an aligner for coupling the housing to a particle sorting system (as described in more detail below). For example, the housing may have 2 or more walls having an aligner for coupling the housing to a particle sorting system, such as 3 or more walls, and including 4 or more walls having an aligner. In certain embodiments, the housing has a wall having an aligner for coupling the housing to a particle sorting system. Each wall having an aligner may include 1 or more aligners, such as 2 or more aligners, such as 3 or more aligners, such as 4 or more aligners, such as 5 or more aligners, such as 7 or more aligners, and including 10 or more aligners. In certain embodiments, the particle sorting device includes an outer wall having 3 aligners.

[0058] Any suitable type of aligner may be used, such as an alignment protrusion, an alignment track, an alignment notch, an alignment groove, an alignment slot, an alignment countersunk hole, an alignment counterbore, an alignment recess, an alignment hole, or a combination thereof. For example, in some cases, the outer wall of the housing includes one or more protrusions, such as a pin, a locating pin, or a bump. In certain embodiments, the aligner is a pin, such as a ball stud. In other cases, the outer wall of the housing includes one or more recesses, such as a hole or a notch. In some cases, the outer wall of the housing includes one or more alignment protrusions and one or more alignment recesses.

[0059] The shape of the aligner can vary, with cross-sectional shapes of interest including, but not limited to: straight cross-sectional shapes, such as square, rectangular, trapezoidal, triangular, hexagonal, etc.; curved cross-sectional shapes, such as circular, elliptical; and irregular shapes, such as a parabolic bottom coupled to a flat top. In some embodiments, the aligner is cylindrical. In other embodiments, the aligner is spherical. In other embodiments, the aligner is polygonal, such as square or rectangular. In certain embodiments, the aligner is a cylindrical pin with a spherical tip (e.g., a ball-head pin). Depending on the coupling register of the particle sorting system, each aligner can have the same or different shapes (as described in more detail below). In some embodiments, each aligner has the same shape. In other embodiments, each aligner has a different shape. In other embodiments, two or more aligners have the same shape, and one or more aligners have different shapes.

[0060] In some embodiments, the outer wall of the shell includes one or more fasteners for maintaining contact between the particle sorting module and the particle sorting system (as described in more detail below). Suitable fasteners may include, but are not limited to, magnets, hook and loop fasteners, Velcro, non-permanent adhesives, or combinations thereof. In certain embodiments, the outer wall of the shell includes one or more magnets for coupling to one or more magnets on a register of the particle sorting system. In these embodiments, the alignment between the particle sorting module and the register can be achieved by coupling the magnets on the register with the magnets on the outer wall of the particle sorting module shell. In some embodiments, the particle sorting module includes an aligner and a magnet. In other embodiments, the aligner includes one or more magnets, such as a protrusion with a magnet at the tip (e.g., a ball pin protrusion). In certain embodiments, the aligner is a magnet, such as a magnetic protrusion, a magnetic ball at the tip of a pin protrusion, or a magnet located within a hole or recess. In this article, the term "magnet" is used in its conventional sense to refer to a magnetic material having a persistent magnetic field so that the magnetic field from the magnet does not substantially decrease over time. For example, the magnet may be an iron alloy material having aluminum, nickel and cobalt (i.e., Alnico magnet), a ceramic or ferrite magnet, a rare earth magnet such as a samarium cobalt magnet (e.g., SmCo5), a neodymium alloy (NdFeB) magnet (e.g., Nd2Fe 14 B) or a combination thereof. The magnetic field generated by the magnet of interest located at the proximal end of the connector ranges from 0.01 T to 10 T, or from 0.01 T to 5 T, or from 0.01 T to 2 T, or from 0.1 T to 2 T, or from 0.1 T to 1.5 T, including from 0.1 T to 1 T, depending on the size of the magnet.

[0061] In certain embodiments, the particle sorting module is a closed particle sorting module, such as those described in U.S. Patent Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosure of which is incorporated herein by reference.

[0062] In certain embodiments, the particle sorting module of interest includes one or more sets of integrated droplet deflectors. The subject droplet deflectors may include 2 or more sets of parallel metal plates, such as 3 or more sets, such as 4 or more sets, such as 5 or more sets, such as 6 or more sets, such as 7 or more sets, such as 8 or more sets, such as 9 or more sets, and including 10 or more sets of parallel metal plates. The parallel metal plates of the subject droplet deflector may be formed of any suitable metal capable of generating an electric field, and may include, but are not limited to, aluminum, brass, chromium, cobalt, copper, gold, indium, iron, lead, nickel, platinum, palladium, tin, steel (e.g., stainless steel), silver, zinc, and combinations and alloys thereof, such as aluminum alloys, aluminum-lithium alloys, aluminum-nickel-copper alloys, aluminum-copper alloys, aluminum-magnesium alloys, aluminum-magnesium oxide alloys, aluminum-silicon alloys, aluminum-magnesium-manganese-platinum alloys, cobalt alloys, cobalt-chromium alloys, cobalt-tungsten alloys, cobalt-molybdenum-carbon alloys, cobalt-chromium-nickel-molybdenum-iron-tungsten alloys, copper alloys, copper-arsenic alloys, copper-beryllium alloys, copper-silver alloys, copper-zinc alloys (e.g., brass), copper-tin alloys (e.g., bronze), copper-nickel alloys, copper-tungsten alloys, copper-gold-silver alloys, copper-nickel-iron alloys, copper-manganese-tin alloys, copper-aluminum-zinc-tin alloys, copper-gold alloys, gold alloys, gold-silver alloys, indium alloys, indium-tin alloys, indium-tin oxide alloys, iron alloys, iron-chromium alloys ( For example, steel), iron-chromium-nickel alloys (such as stainless steel), iron-silicon alloys, iron-chromium-molybdenum alloys, iron-carbon alloys, iron-boron alloys, iron-magnesium alloys, iron-manganese alloys, iron-molybdenum alloys, iron-nickel alloys, iron-phosphorus alloys, iron-titanium alloys, iron-vanadium alloys, lead alloys, lead-antimony alloys, lead-copper alloys, lead-tin alloys, lead-tin-antimony alloys, nickel alloys, nickel-manganese-aluminum-silicon alloys, nickel-chromium alloys, nickel-copper alloys, nickel, molybdenum-chromium-tungsten alloys, nickel-copper-iron-manganese alloys, nickel-carbon alloys, nickel-chromium-iron alloys, nickel-silicon alloys, nickel-titanium alloys, silver alloys, silver-copper alloys (for example, pure silver), silver-copper-germanium alloys (for example, Argentium pure silver), silver-gold alloys, silver-copper-gold alloys, silver-platinum alloys, tin alloys, tin-copper-antimony alloys, tin-lead-copper alloys, tin-lead-antimony alloys, titanium alloys, titanium-vanadium-chromium alloys, titanium-aluminum alloys, titanium-aluminum-vanadium alloys, zinc alloys, zinc-copper alloys, zinc-aluminum-magnesium-copper alloys, zirconium alloys, zirconium-tin alloys, or combinations thereof.

[0063] The parallel metal plates of the subject droplet deflector can be any suitable shape, such as a circle, an ellipse, a semicircle, a crescent, a star, a square, a triangle, a rhombus, a pentagon, a hexagon, a heptagon, an octagon, a rectangle, or other suitable polygon. In some embodiments, the parallel metal plates are rectangular. As described in more detail below, in some cases, the parallel metal plates are twisted, such as a twisted rectangle with a twist angle of 5° or more, such as 10° or more, such as 15° or more, such as 20° or more, such as 25° or more, such as 30° or more, such as 35° or more, such as 40° or more, such as 45° or more, such as 50° or more, such as 55° or more, and including having a twist angle of 60° or more.

[0064] According to the shape of the metal plate, the size can vary. In some embodiments, the width of each metal plate ranges from 0.5mm to 10mm, for example, from 1mm to 9.5mm, for example, from 1.5mm to 9mm, for example, from 2mm to 8.5mm, for example, from 2.5mm to 8mm, for example, from 3mm to 7.5mm, for example, from 3.5mm to 7mm, for example, from 4mm to 6.5mm, and includes a width outside the range of from 4.5mm to 6mm. The length also varies, and its range is from 10mm to 500mm, for example, from 15mm to 450mm, for example, from 20mm to 400mm, for example, from 25mm to 350mm, for example, from 30mm to 300mm, for example, from 35mm to 250mm, for example, from 40mm to 200mm, for example, from 45mm to 150mm, and includes from 50mm to 100mm. In some embodiments, the metal plate is an asymmetric polygon, wherein the width of the first end is less than the width of the second end. The width at each end can range from 0.01mm to 10mm, for example from 0.05mm to 9.5mm, for example from 0.1mm to 9mm, for example from 0.5mm to 8.5mm, for example from 1mm to 8mm, for example from 2mm to 8mm, for example from 2.5mm to 7.5mm, and including from 3mm to 6mm. In some embodiments, the cover strip is an asymmetric polygon having a first end with a width of from 1 to 10mm and a second end with a width of from 2 to 5mm. For example, the cover strip can be an asymmetric polygon having a first end with a width of 5mm and a second end with a width of 10mm. In an embodiment, the surface area of ​​each metal plate can be varied as desired and can range from 0.25 to 15cm 2 , for example 0.5 to 14 cm 2 , for example 0.75 to 13 cm 2 , e.g. 1 to 12 cm 2 , e.g. 1.5 to 11 cm 2 , and includes 2 to 10cm 2 .

[0065] In certain embodiments, the particle sorting module includes one or more droplet deflectors, such as those described in U.S. Patent Publication No. 2018 / 0095022, filed on June 14, 2017, the disclosure of which is incorporated herein by reference.

[0066] In some embodiments, the outer wall of the shell includes one or more electrical connections, which are configured for electrical connection between the particle sorting module and the particle sorting system. For example, the outer shell may include 2 or more electrical connections, such as 3 or more electrical connections, such as 4 or more electrical connections, such as 5 or more electrical connections, and including 10 or more electrical connections. In some embodiments, the electrical connection provides power to the droplet deflector plate. Any convenient electrical connection can be used, such as conductive pins, conductive pads, wires or coils protruding or recessed within the outer wall of the shell. In some embodiments, the particle sorting module of interest includes 5 or more conductive pins.

[0067] Particle sorting system

[0068] Aspects of the present disclosure also include systems for sorting particle components of a sample, such as cells in a biological sample. Systems according to certain embodiments include a particle sorting module having an opening configured for visualizing droplets of a deflected fluid stream as described above, and an imaging sensor configured to capture one or more images through the opening (e.g., a boundary of a sample collection port at a distal end of the particle sorting module).

[0069] In embodiments, the imaging sensor may be any suitable device capable of capturing an optical image and converting it into an electronic data signal, including but not limited to a charge coupled device, 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 some embodiments, the imaging sensor is a CCD camera. For example, the camera may be an electron multiplying CCD (EMCCD) camera or an intensified CCD (ICCD) camera. In other embodiments, the imaging sensor is a CMOS-type camera.

[0070] As desired, the number of imaging sensors in the subject system can vary. For example, the subject system can include one imaging sensor or more, such as two imaging sensors or more, such as three imaging sensors or more, such as four imaging sensors or more, such as five imaging sensors or more, and include ten imaging sensors or more. In some embodiments, the system includes one imaging sensor. In other embodiments, the system includes two imaging sensors. In the case where the system includes more than one imaging sensor, each imaging sensor can be oriented at an angle ranging from 10° to 90° relative to another imaging sensor (based on the XY plane), such as from 15° to 85°, such as from 20° to 80°, such as from 25° to 75°, and including from 30° to 60°. In some embodiments, each imaging sensor is oriented orthogonally to each other (based on the XY plane). For example, in the case where the subject system includes two imaging sensors, the first imaging sensor is oriented orthogonally to the second imaging sensor (based on the XY plane).

[0071] Where the subject system includes more than one imaging sensor, each imaging sensor may be the same sensor or combination of sensors. For example, where the subject system includes two imaging sensors, in some embodiments, the first imaging sensor is a CCD type device and the second imaging sensor is a CMOS type device. In other embodiments, both the first imaging sensor and the second imaging sensor are CCD type devices. In other embodiments, both the first imaging sensor and the second imaging sensor are CMOS type devices.

[0072] In some embodiments, the imaging sensor is stationary, maintaining a single position relative to the opening in the particle sorting module housing. In other embodiments, the imaging sensor can be configured to move, for example, along the path of the deflected fluid flow. For example, the imaging sensor can be configured to move upstream and downstream along the deflected fluid flow, capturing images in multiple detection fields through the opening in the particle sorting module housing. In the case where the imaging sensor is configured to move, the imaging sensor can be displaced continuously. In other embodiments, the imaging sensor can be displaced at discrete intervals, such as in increments of 1 mm or greater, such as in increments of 2 mm or greater, and including increments of 5 mm or greater.

[0073] The imaging sensor may be configured to capture images continuously or at discrete intervals. In some cases, the imaging sensor of interest is configured to capture images continuously. In other cases, the imaging sensor is configured to take measurements at discrete intervals, such as capturing images of the fluid flow every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds, and including every 1000 milliseconds or some other interval.

[0074] As described in more detail below, the imaging sensor is configured to capture one or more images of the deflected fluid flow. For example, the imaging sensor can be configured to capture 2 or more images of the deflected fluid flow, such as 3 or more images, such as 4 or more images, such as 5 or more images, such as 10 or more images, such as 15 or more images, and including 25 or more images. In the case of capturing multiple images, the processor can include a digital imaging processing algorithm for combining multiple images when necessary.

[0075] Depending on the desired image resolution, the imaging sensor may have an exposure time of 100 ms or less, such as 75 ms or less, such as 50 ms or less, such as 25 ms or less, such as 10 ms or less, such as 5 ms or less, such as 1 ms or less, such as 0.1 ms or less, such as 0.01 ms or less, such as 0.001 ms or less, such as 0.0001 ms or less, such as 0.00001 ms or less, and including 0.000001 ms or less when reading out the full sensor. For example, the exposure time of the imaging sensor may range from 0.0001 ms to 10 ms, such as from 0.001 ms to 5 ms, such as from 0.01 ms to 2 ms, and including from 0.1 ms to 1 ms.

[0076] In some embodiments, the imaging sensor in the subject system can have an active pixel of 1M or larger, such as 1.5M or larger, such as 2M or larger, 2.5M or larger, or 3M or larger. In some aspects, the pixel corresponds to an actual physical size of about 0.3 μm. Depending on the detection field, in some cases, the imaging sensor has a 150 mm 2 or larger sensor area, e.g. about 150mm 2 Up to about 175mm 2 , about 175mm 2 Up to about 200mm 2 , 200mm 2 Up to about 225mm 2 , about 225mm 2 Up to about 250mm 2 , about 250mm 2 Up to about 300mm 2 , about 300mm 2 Up to about 400mm 2 , about 400mm 2 Up to about 500mm 2 , about 500mm 2 Up to about 750mm 2 , about 750mm2 Up to about 1000mm 2 or about 1000mm 2 or larger.

[0077] The imaging sensor may be positioned at any suitable distance from the opening in the particle sorting module housing as long as the sensor is capable of capturing an image of the deflected fluid flow through the opening. For example, the imaging sensor may be positioned 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 2.5 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 25 mm or more, and including 50 mm or more from the opening in the particle sorting module housing.

[0078] In some cases, the imaging sensor also includes an optical adjustment component. The optical adjustment component can be configured to change the captured image as needed, such as increasing or reducing the size of the capture or enhancing the optical resolution of the image. In some cases, the optical adjustment is an amplification scheme that is configured to increase the size of the detection field captured by the imaging sensor, such as an increase of 5% or more, such as 10% or more, such as 25% or more, such as 50% or more, and includes increasing the detection field of the imaging sensor by 75% or more. In other cases, the optical adjustment is a de-amplification scheme that is configured to reduce the size of the detection field captured by the imaging sensor, such as a decrease of 5% or more, such as 10% or more, such as 25% or more, such as 50% or more, and includes reducing the size of the detection field captured by the imaging sensor by 75% or more. In some embodiments, the optical adjustment is an enhanced resolution scheme that is configured to increase the resolution of the captured image, such as an increase of 5% or more, such as 10% or more, such as 25% or more, such as 50% or more, and includes increasing the resolution of the captured image by 75% or more. The image captured by the imaging sensor can be adjusted using any convenient optical adjustment scheme, including but not limited to lenses, mirrors, filters, and combinations thereof. In some embodiments, the imaging sensor includes a focusing lens. For example, the focusing lens can be a de-magnifying lens. In other embodiments, the focusing lens is a magnifying lens.

[0079] The imaging sensor of the present disclosure may also include one or more wavelength separators. In this article, the term "wavelength separator" is used in its conventional sense to refer to an optical scheme for separating polychromatic light into its component wavelengths for detection. According to certain embodiments, wavelength separation may include selectively passing or blocking polychromatic light of a specific wavelength or wavelength range. In order to separate light of various wavelengths, the transmitted light may pass through any convenient wavelength separation scheme, including but not limited to colored glass, bandpass filters, interference filters, dichroic mirrors, diffraction gratings, monochromators and combinations thereof, and other wavelength separation schemes. The system may include one or more wavelength separators, such as two or more, such as three or more, such as four or more, such as five or more, and including 10 or more wavelength separators. In one example, the imaging sensor includes a bandpass filter. In another example, the imaging sensor includes two or more bandpass filters. In another example, the imaging sensor includes two or more bandpass filters and a diffraction grating. In yet another example, the imaging sensor includes multiple bandpass filters and a monochromator. In certain embodiments, the imaging sensor includes multiple bandpass filters and a diffraction grating configured as a filter wheel arrangement. Where the imaging sensor includes two or more wavelength separators, the wavelength separators may be used individually or in series to separate polychromatic light into component wavelengths. In some embodiments, the wavelength separators are arranged in series. In other embodiments, the wavelength separators are arranged individually so that one or more measurements are made using each wavelength separator.

[0080] In some embodiments, the system includes one or more optical filters, such as one or more bandpass filters. For example, in some instances, the optical filter of interest is a bandpass filter with a minimum bandwidth ranging from 2nm to 100nm, such as from 3nm to 95nm, such as from 5nm to 95nm, such as from 10nm to 90nm, such as from 12nm to 85nm, such as from 15nm to 80nm, and includes a bandpass filter with a minimum bandwidth ranging from 20nm to 50nm. In other cases, the optical filter is a long pass filter, for example, a long pass filter that attenuates light of a wavelength of 1600 nm or less, for example 1550 nm or less, for example 1500 nm or less, for example 1450 nm or less, for example 1400 nm or less, for example 1350 nm or less, for example 1300 nm or less, for example 1000 nm or less, for example 950 nm or less, for example 900 nm or less, for example 850 nm or less, for example 800 nm or less, for example 750 nm or less, for example 700 nm or less, for example 650 nm or less, for example 600 nm or less, for example 550 nm or less, for example 500 nm or less, and includes a long pass filter that attenuates light of a wavelength of 450 nm or less. In other cases, the optical filter is a short pass filter, such as a short pass filter that attenuates light of a wavelength of 200 nm or greater, such as 250 nm or greater, such as 300 nm or greater, such as 350 nm or greater, such as 400 nm or greater, such as 450 nm or greater, such as 500 nm or greater, such as 550 nm or greater, and includes a short pass filter that attenuates light of a wavelength of 600 nm or greater.

[0081] In other embodiments, the wavelength separator is a diffraction grating. The diffraction grating may include, but is not limited to, a transmission, dispersion, or reflection diffraction grating. The suitable spacing of the diffraction grating may vary according to the configuration of the light source, the detection field, and the imaging sensor, as well as other existing optical adjustment schemes (e.g., focusing lenses), ranging from 0.01 μm to 10 μm, such as from 0.025 μm to 7.5 μm, such as from 0.5 μm to 5 μm, such as from 0.75 μm to 4 μm, such as from 1 μm to 3.5 μm, and including from 1.5 μm to 3.5 μm.

[0082] In some embodiments, each imaging sensor is operably coupled to one or more light sources for illuminating a fluid flow through an opening in a particle sorting module housing. In some embodiments, the light source is a laser, such as a continuous wave laser. For example, the laser can be a diode laser, such as an ultraviolet diode laser, a visible light diode laser, and a near infrared diode laser. In some cases, the diode laser outputs light having a wavelength ranging from 375nm to 1000nm, such as from 405nm to 875nm, such as from 450nm to 800nm, such as from 500nm to 650nm, and including from 525nm to 625nm. In some embodiments, the laser of interest includes a 405nm diode laser. In other embodiments, the laser can be a helium-neon (HeNe) laser. In some cases, the laser is a gas laser, such as a helium-neon laser, an argon laser, a krypton laser, a xenon laser, a nitrogen laser, a CO2 laser, a CO laser, an argon-fluorine (ArF) excimer laser, a krypton-fluorine (KrF) excimer laser, a xenon-chlorine (XeCl) excimer laser, or a xenon-fluorine (XeF) excimer laser, or a combination thereof. In other cases, the subject system includes a dye laser, such as a stilbene, coumarin, or rhodamine laser. In other cases, the laser of interest includes a metal vapor laser, such as a helium-cadmium (HeCd) laser, a helium-mercury (HeHg) laser, a helium-selenium (HeSe) laser, a helium-silver (HeAg) laser, a strontium laser, a neon-copper (NeCu) laser, a copper laser, or a gold laser, and combinations thereof. In other cases, the subject system includes a solid-state laser, such as a ruby ​​laser, a Nd:YAG laser, a NdCrYAG laser, an Er:YAG laser, a Nd:YLF laser, a Nd:YVO4 laser, a Nd:YCa4O(BO3)3 laser, a Nd:YCOB laser, a titanium sapphire laser, a thulium YAG laser, a ytterbium YAG laser, a ytterbium trioxide laser, or a cerium-doped laser, and combinations thereof. In certain embodiments, the light source is a low-power laser (e.g., a 650 nm laser outputting 5 mW).

[0083] In other embodiments, the light source is a non-laser light source, such as a broadband light source, which emits light with a wide wavelength range, for example, the light range spans 50nm or greater, such as 100nm or greater, such as 150nm or greater, such as 200nm or greater, such as 250nm or greater, such as 300nm or greater, such as 350nm or greater, such as 400nm or greater, and includes a range spanning 500nm or greater. For example, a suitable broadband light source emits light with a wavelength from 200nm to 1500nm. Another example of a suitable broadband light source includes a light source that emits light with a wavelength from 400nm to 1000nm. Any convenient broadband light source scheme can be used, such as a halogen lamp, a deuterium arc lamp, a xenon arc lamp, a stable fiber-coupled broadband light source, a broadband LED with a continuous spectrum, a superluminescent light emitting diode, a semiconductor light emitting diode, a wide spectrum LED white light source, a multi-LED integrated white light source, and other broadband light sources or any combination thereof. In some embodiments, the light source for illuminating the fluid flow through the opening in the particle sorting module during image capture includes an infrared LED array.

[0084] In other embodiments, the light source is a narrowband light source that emits a specific wavelength or a narrow range of wavelengths. In some cases, the narrowband light source emits light with a narrow range of wavelengths, and its wavelength range is, for example, 50nm or less, such as 40nm or less, such as 30nm or less, such as 25nm or less, such as 20nm or less, such as 15nm or less, such as 10nm or less, such as 5nm or less, such as 2nm or less, and includes a light source that emits light of a specific wavelength (i.e., monochromatic light). Any convenient narrowband light source scheme can be used, such as a narrow wavelength LED, a laser diode, or a broadband light source coupled to one or more optical bandpass filters, a diffraction grating, a monochromator, or any combination thereof.

[0085] In an embodiment, the imaging sensor is configured to capture an image of an opening through a particle sorting module housing in a detection field. The so-called "detection field" refers to the field of view through the opening in the particle sorting module housing captured by the imaging sensor. The imaged detection field can vary. In some embodiments, the detection field is a deflected fluid flow of a predetermined length. In other embodiments, the detection field is a predetermined portion of the opening in the particle sorting module housing. In some embodiments, the imaging sensor is configured to capture an image of a deflected fluid flow emitted from a droplet deflector in a detection field. In an embodiment, the image detection field can span a deflected fluid flow of 0.001 cm or more, such as 0.005 cm or more, such as 0.01 cm or more, such as 0.05 cm or more, such as 0.1 cm or more, such as 0.5 cm or more, such as 1 cm or more, such as 2 cm or more, such as 5 cm or more, and includes a deflected fluid flow of 10 cm or more.

[0086] In some embodiments, the detection field includes the position of the sample collection port at the distal end of the particle sorting module housing. In some cases, the sample collection port is coupled to a sample collection container, and the imaging sensor is configured to capture the position of the sample collection container at the distal end of the housing. In some embodiments, the imaging sensor is configured to capture an image through an opening of the particle sorting module housing to obtain an imaging opening, and a processor operably coupled to the imaging sensor is configured to calculate the position of the boundary of each sample collection port at the distal end of the housing along the horizontal axis in the imaging opening. In some instances, calculating the position of the boundary includes determining the distance between the boundaries of each sample collection port. For example, the processor can be configured to calculate the boundary of the first sample collection port based on the imaging opening along the horizontal axis from 0.1mm to 100mm, such as from 0.5mm to 95mm, such as from 1mm to 90mm, such as from 1.5mm to 85mm, such as from 2mm to 80mm, such as from 2.5mm to 75mm, and including from 3mm to 70mm. In other cases, the processor may be configured to calculate the boundary of the second sample collection port based on the imaging opening along the horizontal axis from 0.1 mm to 100 mm, such as from 0.5 mm to 95 mm, such as from 1 mm to 90 mm, such as from 1.5 mm to 85 mm, such as from 2 mm to 80 mm, such as from 2.5 mm to 75 mm, and including from 3 mm to 70 mm from the leftmost edge of the image. In other cases, the processor may be configured to calculate the boundary of the third sample collection port based on the imaging opening along the horizontal axis from 0.1 mm to 100 mm, such as from 0.5 mm to 95 mm, such as from 1 mm to 90 mm, such as from 1.5 mm to 85 mm, such as from 2 mm to 80 mm, such as from 2.5 mm to 75 mm, and including from 3 mm to 70 mm.

[0087] In other embodiments, the processor can be configured to calculate that the sample collection ports are spaced 0.1 mm to 10 mm apart from each other along the horizontal axis based on the imaging opening, for example 0.5 mm to 9.5 mm, for example 1 mm to 9 mm, for example 1.5 mm to 8.5 mm, for example 2 mm to 8 mm, for example 2.5 mm to 7.5 mm, and including 3 mm to 7 mm.

[0088] In other embodiments, the processor may be configured to calculate the boundary of the first sample collection port based on the imaging opening along the horizontal axis from 0.1 mm to 100 mm, such as from 0.5 mm to 95 mm, such as from 1 mm to 90 mm, such as from 1.5 mm to 85 mm, such as from 2 mm to 80 mm, such as from 2.5 mm to 75 mm, and including from 3 mm to 70 mm from the rightmost edge of the image. In other cases, the processor may be configured to calculate the boundary of the second sample collection port based on the imaging opening along the horizontal axis from 0.1 mm to 100 mm, such as from 0.5 mm to 95 mm, such as from 1 mm to 90 mm, such as from 1.5 mm to 85 mm, such as from 2 mm to 80 mm, such as from 2.5 mm to 75 mm, and including from 3 mm to 70 mm. In other cases, the processor can be configured to calculate a boundary of the third sample collection port based on the imaging opening along the horizontal axis from 0.1 mm to 100 mm, for example, from 0.5 mm to 95 mm, for example, from 1 mm to 90 mm, for example, from 1.5 mm to 85 mm, for example, from 2 mm to 80 mm, for example, from 2.5 mm to 75 mm, and including from 3 mm to 70 mm from the rightmost edge of the image.

[0089] In some embodiments, the system is configured to capture an image of an opening of a particle sorting module to obtain an imaging opening, and convert the image of the opening into a pixelated image. In this document, the term "pixelated" is used in its conventional sense to indicate that the captured image consists of a plurality of regularly spaced (e.g., grid) pixels, each pixel forming a portion of the captured image. In these embodiments, a processor operably coupled to the imaging sensor can be configured to calculate the pixel position of the boundary of each port at the distal end of the housing along the horizontal axis in the imaging opening.

[0090] In some embodiments, the processor can be configured to calculate the pixel position of the boundary of each sample collection port along the horizontal axis from the leftmost edge of the image based on the imaging opening. For example, the position of the boundary of each sample collection port can be output as the number of pixels along the horizontal axis from the leftmost edge of the image. In some instances, the position of the boundary of each sample collection port along the horizontal axis from the leftmost edge of the image of the opening in the particle sorting module housing can range from 1 pixel to 1000 pixels, such as from 5 pixels to 750 pixels, such as from 10 pixels to 500 pixels, and including from 25 pixels to 250 pixels. In other cases, the position of the boundary of each sample collection port is output as a ratio of the number of pixels passing through the imaging opening along the horizontal axis to the total number of pixels. For example, the total number of pixels passing through the imaging opening can be 100 pixels, and the position of each boundary of the sample collection port can be a ratio ranging from 1 / 100 to 99 / 100, such as from 5 / 100 to 95 / 100, such as from 10 / 100 to 90 / 100, such as from 15 / 100 to 85 / 100, such as from 20 / 100 to 80 / 100, and including a ratio ranging from 25 / 100 to 75 / 100.

[0091] In certain embodiments, a system of interest includes a processor having a memory operably coupled to the processor, wherein the memory includes instructions stored thereon that, when executed by the processor, cause the processor to calculate a total number of pixels passing through an imaging opening along a horizontal axis and multiply the total number of pixels by a predetermined ratio. In these embodiments, the boundaries of each sample collection port are determined based on the imaging opening (e.g., a left boundary and a right boundary along the horizontal axis), and a number of pixels is assigned, and the number of pixels is multiplied by a predetermined ratio. In some cases, the predetermined ratio is calculated by dividing the number of pixels in the imaging opening of each sample collection port boundary by the total number of pixels passing through the imaging opening along the horizontal axis.

[0092] Figure 1 Depicted are example calculations for determining the boundaries of a sample collection port at a distal end of a particle sorting module in accordance with certain embodiments. Figure 1 A particle sorting module 100 is depicted having a droplet deflector 101 that deflects a fluid stream according to trajectories 101a, 101b, and 101c. The particle sorting module 100 includes an opening 102 for visualizing a fluid stream having trajectories 101a, 101b, and 101c. The fluid stream is irradiated with a laser 103. At the distal end of the particle sorting module 100 are sample collection ports 100L, 100C, and 100R.

[0093] The boundary of the first sample collection port (eg, first deflection port) 100L is defined by L l and L rDefinition; the boundary of the second sample collection port (eg, waste port) 100C is defined by C l and C r and the boundary of the third sample collection port (eg, the second deflection port) 100R is defined by R l and R r In this example, the total number of pixels (P) passing through the opening is 100. The sample collection port boundary (L l , L r , C l and C r and R l and R r ) is calculated as P*(pixel position / total pixels).

[0094] In the case where the system is configured to determine the position of the sample collection port based on the imaging opening, the processor may further include a memory having instructions for aligning the fluid stream with one or more sample collection ports. For example, the memory may include instructions for aligning the fluid stream with the center of one of the sample collection ports based on a determined boundary. In one case, the system may be configured to align the fluid stream with the sample collection port. In another case, the system is configured to align the fluid stream with the waste collection port. In yet another case, the system is configured to align the fluid stream along a horizontal axis with the center of the distal end of the housing.

[0095] Figure 2 Depicted is a particle sorting module having an opening configured to visualize droplets of a deflected fluid stream in accordance with certain embodiments. Figure 2 Depicted, the pixel locations of the boundaries of the sample collection port and the trajectories of deflected droplets of fluid flow are determined from captured images through the opening.

[0096] The particle sorting system according to certain embodiments further comprises a sample input subsystem fluidly coupled to an inlet at a proximal end of the particle sorting module. In embodiments, the sample input subsystem is configured to provide a suitable sample flow to a fluid cell nozzle chamber in the particle sorting module. Depending on the desired characteristics of the fluid flow emitted from the fluid nozzle, the sample rate delivered to the particle sorting module through the sample input subsystem can be 1 μL / minute or greater, for example 2 μL / minute or greater, for example 3 μL / minute or greater, for example 5 μL / minute or greater, for example 10 μL / minute or greater, for example 15 μL / minute or greater, for example 25 μL / minute or greater, for example 50 μL / minute or greater, and including 100 μL / minute or greater, wherein, in some cases, the flow rate is 1 μL / second or greater, for example 2 μL / second or greater, for example 3 μL / second or greater, for example 5 μL / second or greater, for example 10 μL / second or greater, for example 15 μL / second or greater, for example 25 μL / second or greater, for example 50 μL / second or greater, and including 100 μL / second or greater.

[0097] In some embodiments, the sample fluid subsystem includes a container, a cover, and one or more ports into the inner cavity of the container. The container has a distal end and a proximal end and a wall between the distal end and the proximal end, which together form an inner cavity within the container. In some embodiments, the outer wall and the inner cavity of the container have the same cross-sectional shape, wherein the cross-sectional shapes of interest include, but are not limited to, curved cross-sectional shapes, such as circular, elliptical; straight cross-sectional shapes, such as square, rectangular, trapezoidal, triangular, hexagonal, etc.; and irregular shapes, such as a parabolic bottom coupled to a flat top. The container can be formed of any suitable material, including, but not limited to, glass, metal, or plastic, such as a soft or hard plastic, polymer, or thermoplastic material as described above.

[0098] The size of the inner cavity of the container can vary depending on the sample size and the size of the particle sorting module, where in some cases the length of the inner cavity of the container can range from 1 cm to 25 cm, such as from 2.5 cm to 22.5 cm, such as from 5 cm to 20 cm, such as from 7.5 cm to 17.5 cm and including from 10 cm to 15 cm, and the width of the inner cavity of the container can range from 1 cm to 20 cm, such as from 2 cm to 17.5 cm, such as from 3 cm to 15 cm, such as from 4 cm to 12.5 cm, and including from 5 cm to 10 cm. In the case where the inner cavity of the container has a cylindrical cross-section, in some embodiments, the diameter can vary, ranging from 1 cm to 10 cm, such as from 2 cm to 9 cm, such as from 3 cm to 8 cm, and including from 4 cm to 7 cm. Therefore, the volume of the container can vary, ranging from 1 to 500 cm 3 , e.g. 5 to 250 cm3 , e.g. 10 to 200 cm 3 , e.g. 15 to 150 cm 3 , e.g. 20 to 125 cm 3 , and includes from 25 to 100cm 3 In some embodiments, the container of the sample input module is a tube having a volume ranging from 1 mL to 500 mL, such as from 2 mL to 400 mL, such as from 3 mL to 300 mL, such as from 4 mL to 200 mL, such as from 5 mL to 150 mL, and including from 10 mL to 100 mL.

[0099] In an embodiment, the container of the sample input module also includes a cover, which is configured to close the proximal end of the container. For example, the cover can be a screw cover, a snap cover or a cover that connects the container by a permanent, semi-permanent or non-permanent adhesive. In some cases, the cover forms a fluid seal with the wall of the container. The cover can be an integral part of the container, including the case where the cover is molded, welded, fused or fixed on the container using a permanent adhesive. In other embodiments, the cover is releasably attached to the container. "Releasably" means that the cover can be freely removed from the proximal end of the container and reattached to the proximal end of the container. In the case where the cover is releasably attached to the container, the cover can be non-permanently fixed to the container by any convenient attachment scheme, including but not limited to: hook and loop fasteners, latches, notches, grooves, pins, ties, hinges, Velcro, non-permanent adhesives, threaded screws or combinations thereof. In some cases, the container includes a threaded outer wall and is threadedly connected to the inner wall of the cover.

[0100] The lid can include one or more ports, such as 2 or more ports, such as 3 or more ports, such as 4 or more ports, entering the inner cavity of the container, and including 5 or more ports. In certain embodiments, the lid includes two ports. These ports can be any convenient ports configured to be connected to the inner cavity fluid of the container or to be connected to the gas. In certain embodiments, the lid includes a port, which is configured to deliver gas to the container to generate a positive pressure within the container, and deliver the sample fluid to the particle sorting module from within the container through a second port. In some cases, the container includes a third opening in the lid to allow air to be discharged.

[0101] Depending on the desired function of the port, any suitable port configuration may be employed, where examples of ports include channels, orifices, channels with check valves, Luer fittings, ports with frangible seals (e.g., disposable ports), and other types of ports. In certain embodiments, the port is configured with a Luer fitting, such as a Luer-Lok or Luer-Slip. The port in the cover of the sample input module may be of any suitable shape, where the cross-sectional shapes of interest include, but are not limited to: straight cross-sectional shapes, such as squares, rectangles, trapezoids, triangles, hexagons, etc.; curved cross-sectional shapes, such as circles, ellipses, etc.; and irregular shapes, such as a parabolic bottom coupled to a plane top. The size of the port may vary, and in some embodiments, ranges from 1 mm to 100 mm, such as from 2 mm to 95 mm, such as from 3 mm to 90 mm, such as from 4 mm to 80 mm, such as from 5 mm to 70 mm, such as from 6 mm to 60 mm, and includes from 10 mm to 50 mm. In some embodiments, the port is a circular orifice and the diameter of the port ranges from 1 mm to 100 mm, such as from 2 mm to 90 mm, such as from 4 mm to 80 mm, such as from 5 mm to 70 mm, such as from 6 mm to 60 mm, and including from 10 mm to 50 mm. Thus, depending on the shape of the port, the port in the cap may have an opening ranging from 0.01 mm to 0.1 mm. 2 Up to 250mm 2 , for example from 0.05mm 2 Up to 200mm 2 , for example from 0.1mm 2 Up to 150mm 2 , for example from 0.5mm 2 Up to 100mm 2 , for example, from 1mm 2 Up to 75mm 2 , for example from 2mm 2 Up to 50mm 2 , and includes from 5mm 2 Up to 25mm 2 .

[0102] In some embodiments, the sample input subsystem includes one or more conduits, which are connected to the inner cavity fluid of the container through one or more ports in the cover. For example, the sample input module may include 2 or more conduits, such as 3 or more conduits, and include 5 or more conduits. Each conduit includes: a proximal end, which contacts the inner cavity of the container; and a distal end, which has an opening for inputting or outputting gas or fluid. In some cases, the sample input subsystem includes: an inlet conduit for delivering gas to the container; and an outlet conduit for delivering the sample fluid from the container to the particle sorting module. In other cases, the sample input subsystem includes two inlet conduits for delivering gas to the container and an outlet conduit for delivering the sample fluid from the container to the particle sorting module.

[0103] Each conduit can have an independently variable length, and each conduit can be 5 cm or greater, such as 7 cm or greater, such as 10 cm or greater, such as 25 cm or greater, such as 30 cm or greater, such as 50 cm or greater, such as 75 cm or greater, such as 100 cm or greater, such as 250 cm or greater, and including 500 cm or greater. The lumen diameter of each conduit can also vary, and can be 0.5 mm or greater, such as 0.75 mm or greater, such as 1 mm or greater, such as 1.5 mm or greater, such as 2 mm or greater, such as 5 mm or greater, such as 10 mm or greater, such as 25 mm or greater, and including 50 mm or greater. For example, depending on the desired flow rate for conveying the fluid from the container to the particle sorting module, the lumen diameter can range from 0.5 mm to 50 cm, such as from 1 mm to 25 mm, and including from 5 mm to 15 mm.

[0104] Each conduit can be configured with one or more valves that can be opened and closed as needed. For example, each conduit can be configured with 2 or more valves, such as 3 or more valves, such as 4 or more valves, and including 5 or more valves. Any convenient valve scheme can be used, including but not limited to: pinch valves, ball valves, butterfly valves, butterfly valves, disc valves, clapper valves, check valves, ball check valves, diaphragm valves, lifting check valves, tilting disc check valves, needle valves, piston valves, plug valves, lifting valves and sliding valves. In some embodiments, the inlet conduit for delivering gas to the container can be configured with two pinch valves. To maintain the sterility of the sample, the inlet conduit from the gas source to the container can include a filter, such as a high efficiency particulate arrestance (HEPA) filter or a filter having pores of 50 μm or less, such as 25 μm or less, such as 15 μm or less, such as 10 μm or less, such as 5 μm or less, such as 1 μm or less, such as 0.5 μm or less, such as 0.1 μm or less, such as 0.05 μm or less, such as 0.01 μm or less, and including 0.001 μm or less.

[0105] In certain embodiments, the system further includes one or more gas sources that are gaseously communicated with the inlet conduit of the sample input subsystem container. In some cases, the gas source is a pressurized gas, such as but not limited to a pressurized gas cylinder, a compressor, etc. In some cases, the pressure that the pressurized gas has is 2psi or greater, such as 5psi or greater, including 10psi or greater, such as 15psi or greater, including 20psi or greater, wherein in some cases, the pressure is 25psi or greater, such as 50psi or greater, or 75psi or greater, including 100psi or greater, or 125psi or greater, such as 150psi or greater. The pressurized gas can be any convenient type of gas that is suitable for generating positive pressure in the container of the sample input subsystem. For example, the pressurized gas can include air, nitrogen, argon, etc.

[0106] In some embodiments, the sample input subsystem includes a sample agitator. Any convenient stirring scheme can be used, including but not limited to ultrasonic oscillators, mechanical or electrical vibrators, eccentric motion devices and other stirring schemes. In certain embodiments, the sample agitator is an eccentric motion instrument with a stepper motor and bearings (for eliminating motion) and an eccentric drive. The sample agitator can be configured to stir the sample input module for any desired duration, such as 1 minute or longer, such as 2 minutes or longer, such as 5 minutes or longer, such as 10 minutes or longer, such as 15 minutes or longer, such as 30 minutes or longer, such as 60 minutes or longer, such as 120 minutes or longer, such as 240 minutes or longer, and including 480 minutes or longer.

[0107] The sample input subsystem may also include a temperature controller, wherein the temperature of the sample in the subject sample input subsystem may be maintained or changed (e.g., increased or decreased) as needed. For example, the temperature controller may be configured to maintain the temperature of the sample input subsystem at from -80°C to 100°C, such as from -75°C to 75°C, such as from -50°C to 50°C, such as from -25°C to 25°C, such as from -10°C to 10°C, and including from 0°C to 25°C. In certain aspects, the subject system includes: a temperature sensor for measuring the temperature within the sample input subsystem container; and a feedback monitor configured to allow the sample input module to operate in a closed-loop manner. For example, in some embodiments, the system may evaluate the temperature in the sample input subsystem, and the feedback monitor may adjust the temperature in the sample input subsystem container (e.g., increase or decrease the temperature) in substantially real time to automatically obtain more effective results as needed.

[0108] The particle sorting system according to certain embodiments also includes: a sheath fluid delivery subsystem, which is used to deliver sheath fluid to the fluid pool nozzle of the particle sorting module. In this article, the term "sheath fluid" is used in its traditional sense to refer to a fluid delivered by a conduit (for example, in a flow cytometer), and the conduit is used to form an annular flow coaxial with the fluid containing the sample, thereby producing a hydrodynamic focusing flow of the sample fluid containing particles in the center of the sheath fluid flow. The sheath fluid of interest can be any convenient buffer composition, such as used in a flow cytometer, and can include one or more salts, including but not limited to potassium phosphate, potassium chloride, sodium phosphate, sodium chloride, preservatives, and chelating agents, such as disodium ethylenediaminetetraacetic acid (EDTA). In an embodiment, the sheath fluid distribution system includes a reservoir containing sheath fluid, a conduit, and the conduit has a proximal end connected to the sheath fluid reservoir fluid and a distal end connected to the sheath fluid input to the particle sorting module.

[0109] In some embodiments, the sheath fluid delivery subsystem includes a pressurized housing with a flexible container having a sheath fluid reservoir positioned within the housing. In other embodiments, the sheath fluid delivery subsystem includes a housing and a first flexible container and a second flexible container positioned in the housing. The first flexible container includes a reservoir and a catheter having a proximal end and a distal end, wherein the proximal end is fluidically coupled to the reservoir and the distal end is configured to couple the catheter to a particle sorting module, and the second flexible container includes a gas reservoir and a port in gaseous communication with the gas reservoir. In these embodiments, the second flexible container is positioned in a housing having the first flexible container and is configured to apply pressure to the reservoir of the first flexible container to deliver the sheath fluid from the distal end of the catheter to the particle sorting module.

[0110] In certain embodiments, the particle sorting system of interest includes a sheath fluid delivery subsystem for delivering sheath fluid to a particle sorting module, such as those described in co-application PCT patent application No. PCT / US2016 / 048433, filed on October 24, 2016 and published as WO 2017 / 040151; U.S. patent application No. 14 / 365,602, now published as U.S. Patent No. 9,551,643; and U.S. Patent Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosures of which are incorporated herein by reference.

[0111] As described above, the subject system is configured to sort the particle components of a sample (e.g., a biological sample). In some embodiments, the system further includes a light detection system configured to irradiate and identify the particle components of the sample in the fluid stream, such as the particle components of the sample in the sample interrogation area of ​​the particle sorting module (e.g., in a test tube). In these embodiments, the system includes: one or more light sources for irradiating the sample in the sample interrogation area of ​​the fluid stream. The light source can be a broadband light source, such as a halogen lamp, a deuterium arc lamp, a xenon arc lamp, a stable fiber-coupled broadband light source, a broadband LED with a continuous spectrum, a superluminescent diode, a semiconductor light emitting diode, a wide spectrum LED white light source, a multi-LED integrated white light source, and other broadband light sources or any combination thereof.

[0112] In other embodiments, the light source is a narrowband light source that emits a specific wavelength or a narrow range of wavelengths, such as a narrow wavelength LED, a laser diode, or a broadband light source coupled to one or more optical bandpass filters, diffraction gratings, monochromators, or any combination thereof.

[0113] In certain embodiments, the light source is a laser. In some cases, the subject system includes a gas laser, such as a helium-neon laser, an argon laser, a krypton laser, a xenon laser, a nitrogen laser, a CO2 laser, a CO laser, an argon-fluorine (ArF) excimer laser, a krypton-fluorine (KrF) excimer laser, a xenon-chlorine (XeCl) excimer laser, or a xenon-fluorine (XeF) excimer laser, or a combination thereof. In other cases, the subject system includes a dye laser, such as a stilbene, coumarin, or rhodamine laser. In other cases, the laser of interest includes a metal vapor laser, such as a helium-cadmium (HeCd) laser, a helium-mercury (HeHg) laser, a helium-selenium (HeSe) laser, a helium-silver (HeAg) laser, a strontium laser, a neon-copper (NeCu) laser, a copper laser, or a gold laser, and combinations thereof. In other cases, the subject system includes a solid-state laser, such as a ruby ​​laser, a Nd:YAG laser, a NdCrYAG laser, an Er:YAG laser, a Nd:YLF laser, a Nd:YVO4 laser, a Nd:YCa4O(BO3)3 laser, a Nd:YCOB laser, a titanium sapphire laser, a thulium YAG laser, an ytterbium YAG laser, an ytterbium trioxide laser, or a cerium-doped laser, and combinations thereof.

[0114] The light source can be positioned at an angle relative to the fluid flow, the angle ranging from 10° to 90°, such as from 15° to 85°, such as from 20° to 80°, such as from 25° to 75°, and including from 30° to 60°. In some embodiments, the light source is positioned at a 90° angle relative to the sample.

[0115] In these embodiments, the system of interest also includes: one or more detectors for detecting and measuring light from the fluid flow. The detectors of interest may include, but are not limited to, optical sensors or photodetectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCDs), enhanced charge coupled devices (ICCDs), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes, phototransistors, quantum dot photoconductors or photodiodes and combinations thereof, and other photodetectors. In certain embodiments, the transmitted light is measured using 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 some embodiments, the imaging sensor is a CCD camera. For example, the camera may be an electron multiplying CCD (EMCCD) camera or an enhanced CCD (ICCD) camera. In other embodiments, the imaging sensor is a CMOS-type camera. In the case of measuring fluorescence or scattered light with a CCD, the effective detection surface area of ​​the CCD may vary, for example, from 0.01 cm 2 Up to 10cm 2 , for example from 0.05cm 2 Up to 9cm 2 , for example, from 0.1cm 2 Up to 8cm 2 , for example from 0.5cm 2 Up to 7cm 2 , and includes from 1cm 2 Up to 5cm 2 The number of photodetectors in the subject system can vary as desired, such as 1 or more, such as 2 or more, such as 3 or more, such as 5 or more, and including 10 or more photodetectors. Where the subject system includes more than one photodetector, each photodetector can be identical, or a collection of two or more photodetectors can be a combination of different photodetectors.

[0116] Depending on the type of irradiation light source and the characteristics of the sample (e.g., the particle size in the sample), the detector can be positioned at a distance from the fluid flow. For example, the detector can be positioned 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 2.5 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 25 mm or more, and including 50 mm or more from the sample. The detector can also be positioned at different angles relative to the sample. For example, the detector can be positioned at an angle relative to the fluid flow, and the angle range is from 10 ° to 90 °, such as from 15 ° to 85 °, such as from 20 ° to 80 °, such as from 25 ° to 75 °, and including from 30 ° to 60 °. In certain embodiments, the detector is positioned at a 90 ° angle relative to the fluid flow. In some embodiments, the system includes: a detector, which is positioned to detect forward scattered light from the fluid flow. In other embodiments, the system includes a detector positioned to detect side scattered light from the fluid flow. In other embodiments, the system includes a detector positioned to detect fluorescence from the fluid flow.

[0117] The particle sorting system according to some embodiments may include a display and an operator input device. For example, the operator input device may be a keyboard, a mouse, etc. The processing module includes a processor that is capable of accessing a memory having instructions stored thereon, which are used to execute the steps of the subject method. The processing module may include an operating system, a graphical user interface (GUI) controller, a system memory, a memory storage device and an input-output controller, a cache, a data backup unit, and many other devices. The processor may be a commercially available processor or one of other processors that are already or will be available. The processor executes the operating system, and the operating system interfaces with the firmware and hardware in a known manner and helps the processor coordinate and execute the functions of various computer programs, which can be written in various programming languages, such as java, Perl, C++, other high-level or low-level languages ​​known in the art, and combinations thereof. The operating system typically cooperates with the processor to coordinate and execute the functions of other components of the computer. The operating system also provides scheduling, input-output control, file and data management, memory management, and communication control and related services, all of which are in accordance with known techniques. The processor may be any suitable analog or digital system. In some embodiments, the processor includes analog electronics that provide feedback control (e.g., negative feedback control).

[0118] The system memory can be any of various known or future memory storage devices. Examples include any commonly used random access memory (RAM), magnetic media such as resident hard disks or tapes, optical media such as read-write optical disks, flash memory devices, or other memory storage devices. The memory storage device can be any of various known or future devices, including optical disk drives, tape drives, removable hard disk drives, or floppy disk drives. This type of memory storage device is usually read and / or written from a program storage medium ((not shown) such as an optical disk, a tape, a removable hard disk, or a floppy disk), respectively. Any of these program storage media, or other media that are currently being used or may be developed later, can be considered a computer program product. As will be understood, these program storage media usually store computer software programs and / or data. Computer software programs (also referred to as computer control logic) are usually stored in system memory and / or program storage devices used with memory storage devices.

[0119] In some embodiments, a computer program product is described that includes a computer usable medium having control logic (computer software program, including program code) stored therein. The control logic, when executed by a processor and a computer, causes the processor to perform the functions described herein. In other embodiments, some functions are primarily implemented in hardware using, for example, a hardware state machine. It will be apparent to a person skilled in the relevant art that a hardware state machine is implemented to perform the functions described herein.

[0120] The memory may be any suitable device in which the processor can store and retrieve data, such as a magnetic, optical or solid-state storage device (including a magnetic or optical disk, a tape or a RAM or any other suitable device, whether fixed or portable). The processor may include a general-purpose digital microprocessor appropriately programmed from a computer-readable medium carrying the necessary program code. The program may be provided to the processor remotely via a communication channel, or the program may be pre-stored in a computer program product using any of those devices associated with the memory, such as a memory or some other portable or fixed computer-readable storage medium. For example, a magnetic or optical disk may carry the program and may be read by a disk writer / reader. The system of the present invention also includes a program, such as an algorithm for implementing the above method in the form of a computer program product. The program according to the present invention may be recorded on a computer-readable medium, such as any medium that can be directly read and accessed by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy disks, hard disk storage media and tapes; optical storage media, such as CD-ROMs; electrical storage media, such as RAM and ROMs; portable flash drives; and hybrid media such as magnetic / optical storage media.

[0121] The processor may also access a communication channel to communicate with a user at a remote location. A remote location means that the user is not in direct contact with the system and the input information is forwarded to the input manager from an external device such as a computer connected to a wide area network ("WAN"), a telephone network, a satellite network, or any other suitable communication channel including a mobile phone (i.e., a smartphone).

[0122] In some embodiments, a system according to the present disclosure may be configured to include a communication interface. In some embodiments, the communication interface includes a receiver and / or a transmitter for communicating with a network and / or another device. The communication interface may be configured for wired or wireless communication, including but not limited to radio frequency (RF) communication (e.g., radio frequency identification (RFID)), Zigbee communication protocol, WiFi, infrared, wireless universal serial bus (USB), ultra-wideband (UWB), Communication protocols and cellular communications, such as Code Division Multiple Access (CDMA) or Global System for Mobile Communications (GSM).

[0123] In one embodiment, the communication interface is configured to include one or more communication ports, e.g., physical ports or interfaces such as a USB port, an RS-232 port, or any other suitable electrical connection port, to allow data communications between the subject system and other external devices such as a computer terminal (e.g., in a doctor's office or hospital environment) configured for similar supplemental data communications.

[0124] In one embodiment, the communication interface is configured for infrared communication, communications or any other suitable wireless communications protocol to enable the subject system to communicate with other devices such as computer terminals and / or networks, communications-enabled mobile phones, personal digital assistants, or any other communications devices that a user may use in conjunction with the subject system.

[0125] In one embodiment, the communication interface is configured to provide a connection for data transmission using the Internet Protocol (IP) through a cellular network, a short message service (SMS), a wireless connection to a personal computer (PC) on a local area network (LAN) connected to the Internet, or a WiFi connection connected to the Internet via a WiFi hotspot.

[0126] In one embodiment, the subject system is configured to communicate via a communication interface (e.g., using a communication interface such as 802.11 or The server device may be another portable device, such as a smart phone, a personal digital assistant (PDA), or a laptop computer; or a larger device, such as a desktop computer, a device, etc. In some embodiments, the server device has: a display, such as a liquid crystal display (LCD); and an input device, such as a button, a keyboard, a mouse, or a touch screen.

[0127] In some embodiments, the communication interface is configured to automatically or semi-automatically communicate data stored in the subject system (eg, in an optional data storage unit) with a network or server device using one or more of the communication protocols and / or mechanisms described above.

[0128] The output controller may include a controller for any of a variety of known display devices, which are used to present information to a user (whether human or machine), whether local or remote. If one of the display devices provides visual information, the information can generally be logically and / or physically organized as an array of picture elements. A graphical user interface (GUI) controller may include any of a variety of known or future software programs for providing a graphical input and output interface between the system and the user, and for processing user input. The functional elements of the computer may communicate with each other via a system bus. In alternative embodiments, some of these communications may be accomplished using a network or other types of remote communications. The output manager may also provide information generated by the processing module to a user at a remote location according to known techniques (e.g., via the Internet, telephone, or satellite network). The presentation of data by the output manager may be implemented according to various known techniques. For example, the data may include SQL, HTML, or XML documents, emails or other files, or other forms of data. The data may include an Internet URL address so that the user can retrieve additional SQL, HTML, XML, or other documents or data from a remote source. The platform or platforms present in the subject system may be any type of known computer platform or type to be developed in the future, although they will typically be a type of computer commonly referred to as a server. However, they may also be mainframe computers, workstations, or other computer types. They may be connected via any known or future type of cables or other communication systems (including wireless systems, whether networked or otherwise). They may be co-located or physically separated. A variety of operating systems may be employed on any computer platform, depending on the type and / or brand of computer platform selected. Suitable operating systems include Windows 10, Windows WindowsXP, Windows 7, Windows 8, iOS, Sun Solaris, Linux, OS / 400, Compaq Tru64 Unix, SGIIRIX, Siemens Reliant Unix, Ubuntu, Zorin OS, and other operating systems.

[0129] Aspects of the present disclosure further include a flow cytometer system having one or more particle sorting modules having an opening for visualizing a deflected fluid stream as described above. Suitable flow cytometric systems and methods for sorting particles in a sample include, but are not limited to, those described in Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford University Press (1997); Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology 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, the disclosures of which are incorporated herein by reference. In some cases, flow cytometer systems of interest include the BD Biosciences FACSCanto TM II flow cytometer, BD Accuri TM Flow cytometer, BD Biosciences FACSCelesta TM Flow cytometer, BD Biosciences FACSLyric TM Flow cytometer, BD Biosciences FACSVerse TM Flow cytometer, BD Biosciences FACSymphony TM Flow cytometer, BD Biosciences LSR Fortessa TM Flow cytometer, BD Biosciences LSRFortess TMX-20 Flow Cytometer and BD Biosciences FACSCalibur TM Cell sorter, BD Biosciences FACSCount TM Cell sorter, BD Biosciences FACSLyric TM Cell Sorter and BD Biosciences Via TM Cell sorter, BD Biosciences Influx TM Cell sorter, BD Biosciences Jazz TM Cell sorter, BD BiosciencesAria TM Cell Sorter and BD Biosciences FACSMelody TM Cell sorting machine, etc.

[0130] In some embodiments, the subject particle sorting system is a flow cytometer system, such as U.S. Pat. Nos. 9,952,076; 9,933,341; 9,726,527; 9,453,789; 9,200,334; 9,097,640; 9,095,494; 9,092,034; 8,975,595; 8,753,573; 8,233,146; 8,140, those systems described in 300; 7,544,326; 7,201,875; 7,129,505; 6,821,740; 6,813,017; 6,809,804; 6,372,506; 5,700,692; 5,643,796; 5,627,040; 5,620,842; 5,602,039; the disclosures of which are incorporated herein by reference in their entirety.

[0131] In some embodiments, the particle sorting system of interest is configured to sort particles with an enclosed particle sorting module, such as those described in U.S. Patent Publication No. 2017 / 0299493 filed on March 28, 2017, the disclosure of which is incorporated herein by reference. For example, the particle sorting system of interest may include a register configured to couple with an aligner on a housing of the particle sorting module. The register may include one or more aligners that are complementary to the aligners on the housing of the particle sorting module. In addition, the particle sorting system may include: one or more fasteners that are used to maintain physical contact between the particle sorting module and the particle sorting system, for example by coupling one or more magnets on the housing of the particle sorting module with one or more magnets on the particle sorting system.

[0132] In certain embodiments, the subject particle system is a flow cytometer system with an excitation module that uses radio frequency multi-channel excitation to generate multiple frequency-shifted beams. In these embodiments, the laser generator may include multiple lasers and one or more acousto-optic components (e.g., acousto-optic deflectors, acousto-optic frequency shifters) to generate multiple frequency-shifted comb beams. In certain cases, the subject system is a flow cytometer system with a laser excitation module, as described in U.S. Patent Nos. 9,423,353 and 9,784,661 and U.S. Patent Publication Nos. 2017 / 0133857 and 2017 / 0350803, the disclosures of which are incorporated herein by reference.

[0133] Method for aligning a deflected fluid stream with a sample collection port

[0134] Aspects of the present disclosure also include methods for aligning a fluid stream with one or more sample collection ports at a distal end of a particle sorting module (as described above). Methods according to certain embodiments include capturing an image through an opening in a particle sorting module, the opening being configured to: visualize droplets of a fluid stream; calculate the pixel position of the boundary of each port at the distal end along a horizontal axis in the imaging opening; and align the fluid stream with the port based on the calculated pixel position of the boundary of each port in the imaging opening. In an embodiment, the subject method can be fully automated, such as reducing or completely eliminating the need for user input or manual determination of the location of the boundaries of the sample collection ports and the alignment of the deflected fluid stream with the sample collection ports. In other words, determining the location of each sample collection port boundary and the alignment of the deflected fluid stream with the sample collection port can require little to no human intervention or manual user input.

[0135] In implementation methods according to certain embodiments, one or more images are captured in a detection field through an opening in a particle sorting module housing. The so-called "detection field" refers to the field of view captured by an imaging sensor through an opening in a particle sorting module housing. The imaging detection field may vary. In some embodiments, the detection field is a predetermined length of the deflected fluid flow. In other embodiments, the detection field is a predetermined portion of the opening in the particle sorting module housing. In some embodiments, the imaging sensor is configured to capture an image of the deflected fluid flow emitted from a droplet deflector in the detection field. In an embodiment, the image detection field may span a deflected fluid flow of 0.001 cm or more, such as 0.005 cm or more, such as 0.01 cm or more, such as 0.05 cm or more, such as 0.1 cm or more, such as 0.5 cm or more, such as 1 cm or more, such as 2 cm or more, such as 5 cm or more, and includes a deflected fluid flow of 10 cm or more.

[0136] While capturing one or more images through an opening in the particle sorting module housing, the detection field is illuminated with a light source. In some embodiments, the detection field is illuminated with a broadband light source or with a narrowband light. Suitable broadband light source schemes may include, but are not limited to, halogen lamps, deuterium arc lamps, xenon arc lamps, stable fiber-coupled broadband light sources, broadband LEDs with continuous spectra, super luminescent diodes, semiconductor light emitting diodes, wide spectrum LED white light sources, multi-LED integrated white light sources, and other broadband light sources or any combination thereof. Suitable narrowband light sources include, but are not limited to, lasers, narrow wavelength LEDs, laser diodes, or broadband light sources coupled to one or more optical bandpass filters, diffraction gratings, monochromators, or any combination thereof.

[0137] Capturing one or more images through an opening in the particle sorting module housing may include illuminating with a combination of light sources, such as with two or more light sources, such as three or more light sources, such as four or more light sources, and including five or more light sources. In the case where the image is captured by illuminating with two light sources, the subject method may include illuminating with two light sources at the same time. In other embodiments, capturing an image may include illuminating with two light sources in sequence. In the case where the two light sources illuminate in sequence, the time that each light source illuminates may independently be 0.001 seconds or longer, such as 0.01 seconds or longer, such as 0.1 seconds or longer, such as 1 second or longer, such as 5 seconds or longer, such as 10 seconds or longer, such as 30 seconds or longer, and including 60 seconds or longer. In embodiments where images are captured by illuminating with two or more light sources in sequence, the duration of illumination of each light source may be the same or different.

[0138] The image of the opening through the particle sorting module housing can be captured continuously or at discrete intervals. In some cases, the method includes capturing images continuously. In other cases, the method includes capturing images at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds and including capturing images of the fluid flow every 1000 milliseconds or some other intervals. One or more images, such as 2 or more images, such as 3 or more images, such as 4 or more images, such as 5 or more images, such as 10 or more images, such as 15 or more images, and including 25 or more images can be captured in the detection field.

[0139] As long as a usable image is captured, an image through the opening of the particle sorting module can be captured at any suitable distance (e.g., from the deflected fluid flow). For example, an image is captured through an opening in the particle sorting module housing at a distance of 0.01 mm or more from the deflected fluid flow, 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 2.5 mm or more, such as 5 mm or more, such as 10 mm or more, such as 15 mm or more, such as 25 mm or more, and including 50 mm or more from the deflected fluid flow. Images can be captured at any angle through the opening. For example, an image can be captured at an angle relative to the axis of the deflected fluid flow, the angle range being from 10° to 90°, such as from 15° to 85°, such as from 20° to 80°, such as from 25° to 75°, and including from 30° to 60°. In some embodiments, the image is captured at a 90° angle relative to the axis of the deflected fluid flow.

[0140] In some embodiments, the method includes calculating the position of the boundary of each sample collection port at the distal end of the shell along the horizontal axis based on the image. In some instances, calculating the position of the boundary includes determining the distance between the boundaries of each sample collection port. For example, the method may include: based on the imaging opening, the boundary of the first sample collection port is calculated along the horizontal axis from 0.1mm to 100mm, such as from 0.5mm to 95mm, such as from 1mm to 90mm, such as from 1.5mm to 85mm, such as from 2mm to 80mm, such as from 2.5mm to 75mm, and includes from 3mm to 70mm. In other cases, the method may include: based on the imaging opening, the boundary of the second sample collection port is calculated along the horizontal axis from the leftmost edge of the image from 0.1mm to 100mm, such as from 0.5mm to 95mm, such as from 1mm to 90mm, such as from 1.5mm to 85mm, such as from 2mm to 80mm, such as from 2.5mm to 75mm, and includes from 3mm to 70mm. In other cases, the method may include: based on the imaging opening calculation, the boundary of the third sample collection port is from 0.1mm to 100mm along the horizontal axis from the leftmost edge of the image, for example, from 0.5mm to 95mm, for example, from 1mm to 90mm, for example, from 1.5mm to 85mm, for example, from 2mm to 80mm, for example, from 2.5mm to 75mm, and including from 3mm to 70mm.

[0141] In other embodiments, the method includes calculating the space between each sample collection port (i.e., the distance between the boundaries of each sample collection port). For example, the method may include: based on the imaging opening, the sample collection ports are spaced 0.1 mm to 10 mm, such as 0.5 mm to 9.5 mm, such as 1 mm to 9 mm, such as 1.5 mm to 8.5 mm, such as 2 mm to 8 mm, such as 2.5 mm to 7.5 mm, and including 3 mm to 7 mm from each other along the horizontal axis.

[0142] In some cases, the method includes: based on the imaging opening, the boundary of the first sample collection port is calculated from 0.1mm to 100mm along the horizontal axis from the rightmost edge of the image, such as from 0.5mm to 95mm, such as from 1mm to 90mm, such as from 1.5mm to 85mm, such as from 2mm to 80mm, such as from 2.5mm to 75mm, and including from 3mm to 70mm. In other cases, the method includes: based on the imaging opening, the boundary of the second sample collection port is calculated from 0.1mm to 100mm along the horizontal axis from the rightmost edge of the image, such as from 0.5mm to 95mm, such as from 1mm to 90mm, such as from 1.5mm to 85mm, such as from 2mm to 80mm, such as from 2.5mm to 75mm, and including from 3mm to 70mm. In other cases, the method includes: based on the imaging opening, the boundary of the third sample collection port is calculated along the horizontal axis from 0.1mm to 100mm, for example, from 0.5mm to 95mm, for example, from 1mm to 90mm, for example, from 1.5mm to 85mm, for example, from 2mm to 80mm, for example, from 2.5mm to 75mm, and including from 3mm to 70mm from the rightmost edge of the image.

[0143] In some embodiments, the method includes generating a pixelated image. In some cases, any suitable image generation scheme can be used, wherein the pixelated image can be output in a digital (e.g., editable) format. In these embodiments, the method may include calculating the pixel position of the boundary of each port at the distal end along the horizontal axis in the imaging opening. In some cases, the method includes: calculating the pixel position of the boundary of each sample collection port along the horizontal axis from the leftmost edge of the image based on the imaging opening. In other cases, the method includes: calculating the pixel position of the boundary of each sample collection port along the horizontal axis from the rightmost edge of the image based on the imaging opening. In some cases, the method includes outputting the number of pixels along the horizontal axis from one or both of the leftmost edge or the rightmost edge of the image. For example, the position of the boundary of each sample collection port along the horizontal axis from the leftmost edge or the rightmost edge of the image of the opening in the particle sorting module housing can range from 1 pixel to 1000 pixels, such as from 5 pixels to 750 pixels, such as from 10 pixels to 500 pixels, and including from 25 pixels to 250 pixels. In other cases, the position of the boundary of each sample collection port is output as a ratio of the number of pixels passing through the imaging opening along the horizontal axis to the total number of pixels. For example, the total number of pixels passing through the imaging opening can be 100 pixels, and the position of each boundary of the sample collection port can be a ratio ranging from 1 / 100 to 99 / 100, such as from 5 / 100 to 95 / 100, such as from 10 / 100 to 90 / 100, such as from 15 / 100 to 85 / 100, such as from 20 / 100 to 80 / 100, and including a ratio ranging from 25 / 100 to 75 / 100.

[0144] In some embodiments, calculating the pixel position of each border of the sample collection port includes calculating the total number of pixels along the horizontal axis passing through the imaging opening; and multiplying the total number of pixels by a predetermined ratio. In these embodiments, the border of each sample collection port (e.g., the left border and the right border along the horizontal axis) is determined based on the imaging opening and is assigned a pixel number, and the pixel number is multiplied by a predetermined ratio.

[0145] In certain embodiments, the method includes aligning the fluid stream with the sample collection port based on the calculated pixel position of the boundary of each sample collection port at the distal end of the particle sorting module housing. In some instances, one or more deflected fluid streams are aligned with the sample collection port. In other cases, the non-deflected fluid stream (i.e., the longitudinal axis of the fluid stream) is aligned with the center of the distal end of the particle sorting module. In one example, the non-deflected fluid stream can be aligned with the waste collection port at the distal end of the particle sorting module.

[0146] As described above, the fluid stream can be deflected by subjecting the fluid stream to a deflection force with a droplet deflector. In order to align the deflected fluid stream with the sample collection port having a position determined as above, the voltage applied to the metal plate of the droplet deflector can be adjusted as needed. The voltage applied to the deflector plate to deflect the charged particles can be 10mV or greater, such as 25mV or greater, such as 50mV or greater, such as 100mV or greater, such as 250mV or greater, such as 500mV or greater, such as 750mV or greater, such as 1000mV or greater, such as 2500mV or greater, such as 5000mV or greater, such as 10000V or greater, such as 15000V or more, such as 25000V or greater, such as 50000V or greater, and including 100000V or greater. In some embodiments, the voltage applied to the deflector plates is from 0.5 kV to 15 kV, such as from 1 kV to 15 kV, such as from 1.5 kV to 12.5 kV, and including from 2 kV to 10 kV. Thus, the electric field strength between the deflector plates ranges from 0.1 V / m to 1x10 7 V / m, for example from 0.5V / m to 5x10 6 V / m, for example, from 1V / m to 1x10 6 V / m, for example from 5V / m to 5x10 5 V / m, for example, from 10V / m to 1x10 5 V / m, and includes from 50V / m to 5x10 4 V / m, e.g. 1x10 5 V / m to 2x10 6 V / m.

[0147] In some embodiments, the method includes sorting the components of the sample with a subject particle sorting module. In some cases, the sample is a biological sample, and the method includes sorting and collecting two or more different types of cells. In some embodiments, the sample is a biological sample. The term "biological sample" refers to a subset, a cell of a whole organism, plant, fungus or animal tissue in its conventional sense, or in some cases can be a component found in blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, amniotic fluid, amniotic umbilical cord blood, urine, vaginal fluid and semen. Therefore, "biological sample" refers to: a subset of a protist or its tissue, and a homogenate, lysate or extract prepared from a subset of an organism or its tissue, including but not limited to, for example, plasma, serum, cerebrospinal fluid, lymph, skin slices, respiratory tract, gastrointestinal tract, cardiovascular and urogenital tract, tears, saliva, milk, blood cells, tumors, organs. A biological sample can be any type of organism tissue, including healthy tissue and diseased tissue (e.g., cancer, malignancy, necrosis, etc.). In certain embodiments, the biological sample is a liquid sample, such as blood or a derivative thereof, such as plasma, tears, urine, semen, etc., wherein in some cases, the sample is a blood sample, including whole blood, such as blood obtained from a venipuncture or finger stick (wherein the blood may or may not be combined with any reagents, such as preservatives, anticoagulants, etc., prior to testing).

[0148] In certain embodiments, the source of the sample is "mammal" or "mammalian", wherein these terms are widely used to describe organisms belonging to the mammal class, including Carnivora (e.g., dogs and cats), Rodentia (e.g., mice, guinea pigs and rats) and Primates (e.g., humans, chimpanzees and monkeys). In some cases, the subject is a human. The method can be applied to samples obtained from human subjects of both sexes and at any stage of development (i.e., newborns, infants, teenagers, adolescents, adults), wherein in certain embodiments, the human subject is a teenager, adolescent or adult. Although the present invention can be applied to samples from human subjects, it should be understood that the method can also be performed on samples from other animal subjects (i.e., in "non-human subjects"), such as, but not limited to, birds, mice, rats, dogs, cats, livestock and horses.

[0149] The cells of interest can be targeted for separation from a fluid stream according to various parameters (e.g., phenotypic features identified by attaching specific fluorescent markers to the cells of interest). In certain embodiments, the system is configured to deflect analytical droplets determined to include target cells. The subject method can be used to target a variety of cells for sorting. The 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 erythroid cells. The target cells of interest include cells with convenient cell surface markers or antigens that can be captured or labeled by convenient affinity agents or their conjugates. For example, the target cell may include cell surface antigens such as CD11b, CD123, CD14, CD15, CD16, CD19, CD193, CD2, CD25, CD27, CD3, CD335, CD36, CD4, CD43, CD45RO, CD56, CD61, CD7, CD8, CD34, CD1c, CD23, CD304, CD235a, T cell receptor α / β, T cell receptor γ / δ, CD253, CD95, CD20, CD105, CD117, CD120b, Notch4, Lgr5 (N-terminal), SSEA-3, TRA-1-60 antigen, biganglioside 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 cord blood.

[0150] The method for sorting the components of a sample according to some embodiments includes introducing a fluid sample with target particles into a particle sorting module fluid nozzle. When leaving the fluid nozzle, the particles pass through the sample interrogation area substantially one at a time, wherein each particle is irradiated by a light source, and the measurement of light scattering parameters and fluorescence emission as required is recorded for each particle (e.g., two or more light scattering parameters and one or more fluorescence emission measurements). The particles pass through the sample interrogation area in the particle sorting module substantially one at a time in the fluid flow, wherein each particle is illuminated by a light source. Depending on the characteristics of the fluid flow being interrogated, a fluid flow of 0.001 mm or more, such as 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, and including a fluid flow of 1 mm or more can be irradiated with light. In certain embodiments, the method includes, for example, irradiating a planar cross section of the fluid flow in the sample interrogation area with a laser (as described above). In other embodiments, methods include irradiating a predetermined length of the fluid stream in a sample interrogation region, eg, corresponding to an irradiation profile of a diffuse laser beam or lamp.

[0151] A detector, such as a photomultiplier tube (or "PMT"), connected in series with the sensing region is used to record light passing through each particle (in some cases referred to as forward light scatter), light reflected orthogonally to the direction of flow of particles through the sensing region (in some cases referred to as orthogonal light scatter or side light scatter), and fluorescence emitted from the particle (if labeled with a fluorescent marker) as the particle passes through the sensing region and is illuminated by the energy source. Each of the forward light scatter (or FSC), orthogonal light scatter (SSC), and fluorescence emission (FL1, FL2, etc.) comprises a separate parameter for each particle (or each "event"). Thus, for example, two, three, or four parameters may be collected (and recorded) from particles labeled with two different fluorescent markers.

[0152] As described above, suitable light detection schemes include, but are not limited to, optical sensors or photodetectors, such as active pixel sensors (APS), avalanche photodiodes, image sensors, charge coupled devices (CCD), enhanced charge coupled devices (ICCD), light emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photovoltaic cells, photodiodes, photomultiplier tubes, phototransistors, quantum dot photoconductors or photodiodes and combinations thereof, and other photodetectors. In some embodiments, a charge coupled device (CCD), a semiconductor charge coupled device (CCD), an active pixel sensor (APS), a complementary metal oxide semiconductor (CMOS) image sensor, or an N-type metal oxide semiconductor (NMOS) image sensor is used to measure light from an irradiated fluid stream at a sample interrogation region of a particle sorting module. In some embodiments, light is measured using a charge coupled device (CCD). In the case of using a CCD to measure light from an irradiated fluid stream at a sample interrogation region of a particle sorting module, the effective detection surface area of ​​the CCD can vary, for example, from 0.01 cm 2 Up to 10cm 2 , for example from 0.05cm 2 Up to 9cm 2 , for example, from 0.1cm 2 Up to 8cm 2 , for example from 0.5cm 2 Up to 7cm 2 , and includes from 1cm 2 Up to 5cm 2 .

[0153] The method in certain embodiments also includes data acquisition, analysis and recording, for example, using a computer, wherein multiple data channels record data from each detector about light scattering and fluorescence emitted when each particle passes through the sample interrogation area of ​​the particle sorting module. In these embodiments, the analysis includes classifying and counting the particles so that each particle appears as a set of digitized parameter values. The subject system can be set to trigger on a selected parameter to distinguish particles of interest from background and noise. "Trigger" refers to a preset threshold for detecting a parameter and can be used as a means of detecting the passage of particles through a light source. Detection of an event exceeding a selected parameter threshold will trigger the acquisition of light scattering and fluorescence data for the particles. Data for particles or other components in the analysis medium that cause a response below the threshold is not acquired. The trigger parameter can be the detection of forward scattered light caused by the particle passing through the light beam. The flow cytometer then detects and collects light scattering and fluorescence data for the particles.

[0154] Then, based on the data collected for the entire population, the specific sub-subgroups of interest are further analyzed by "gating". In order to select an appropriate gate, the data is plotted to obtain the best possible subgroup separation. This procedure can be performed by plotting forward light scatter (FSC) and side (i.e., orthogonal) light scatter (SSC) on a two-dimensional dot plot. Then a subgroup of particles (i.e., those cells within the gate) is selected, and particles that are not within the gate are excluded. If necessary, the gate can be selected by drawing a line around the desired subgroup using a cursor on the computer screen. Those particles within the gate are then further analyzed by plotting other parameters of these particles (e.g., fluorescence). If necessary, the above analysis can be configured to generate counts of particles of interest in the sample.

[0155] In some cases, the particle sorting module is a closed particle sorting module having one or more aligners for coupling to a particle sorting system as described above. In these embodiments, in order to couple the particle sorting module to the particle sorting system, the aligners on the outer wall of the particle sorting module housing are placed in contact with the aligners on the register of the particle sorting system. When present, one or more fasteners may engage when the aligners on the outer wall of the particle sorting module housing contact the aligners of the particle sorting system register to secure the particle sorting module to the particle sorting system. After the particle sorting module is coupled to the particle sorting system, a certain amount of fluid sample is then injected into the particle sorting module for analysis and cell sorting as described above.

[0156] In certain embodiments, a method for sorting components of a sample includes sorting particles (e.g., cells in a biological sample) using a closed particle sorting module, such as described in U.S. Patent Publication No. 2017 / 0299493, filed on March 28, 2017, the disclosure of which is incorporated herein by reference.

[0157] Kits

[0158] Aspects of the invention further include kits, wherein the kit includes one or more particle sorting modules as described herein. In some embodiments, the kit includes a particle sorting module having: an opening configured for visualization of droplets of a deflected fluid stream, and one or more sample collection containers for coupling to the particle sorting module. In some embodiments, the kit includes a droplet deflector, such as a set of parallel metal plates and instructions for mounting the droplet deflector into the particle sorting module. In some instances, the kit includes one or more connectors for coupling the particle sorting module to a sample fluid delivery system and a sheath fluid delivery system.

[0159] The various components of the kit may be present in separate containers, or some or all of them may be pre-assembled. For example, in some cases, one or more components of the kit (e.g., each particle sorting module, sample collection container, droplet deflector) are present in a sealed bag, such as a sterile foil bag or envelope.

[0160] In addition to the above-mentioned components, the theme kit may further include (in certain embodiments) instructions for practicing the theme method. These instructions can be present in the theme kit in various forms, wherein one or more forms may be present in the kit. One form of these instructions can be present in the packaging of the kit, a packaging insert, etc. as information printed on a suitable medium or substrate (e.g., one or more sheets of printed information). Another form of these instructions is a computer-readable medium, such as a floppy disk, a compact disc (CD), a portable flash drive, etc., on which information is recorded. Another form in which these instructions can exist is a website, which can be used via the Internet to access information at a removed site.

[0161] use

[0162] The present subject matter, particle sorting modules, particle sorting systems, methods, and computer systems can be used in a variety of applications where it is desirable to analyze and sort particle components in a sample (e.g., a biological sample) in a fluid medium. Embodiments of the present disclosure can be used where it is desirable to provide a flow cytometer with improved cell sorting accuracy, enhanced particle collection, particle charging efficiency, more precise particle charging, and enhanced particle deflection during cell sorting.

[0163] Embodiments of the present disclosure may also be used in applications where cells prepared from biological samples may be desired for research, laboratory testing, or for treatment. In certain embodiments, subject methods and devices may facilitate obtaining single cells prepared from target fluids or tissue biological samples. For example, subject methods and systems facilitate obtaining cells from fluids or tissue samples to be used as research or diagnostic samples for diseases such as cancer. Similarly, subject methods and systems may facilitate obtaining cells for treatment from fluids or tissue samples. Compared to conventional flow cytometer systems, the methods and devices of the present disclosure allow separation and collection of cells from biological samples (e.g., organs, tissues, tissue fragments, liquids) with higher efficiency and lower cost. In certain embodiments, subject particle sorting modules are closed, and the subject particle sorting modules provide increased sterility to the particle sorting system, which enhances the collection of samples of higher purity, and reduces the incidence of cross contamination between analyzed samples, such as in research and high-throughput laboratory testing.

[0164] Notwithstanding the appended claims, the present disclosure is defined by the following clauses:

[0165] 1. A particle sorting module comprising an opening configured to visualize droplets of a deflected fluid stream.

[0166] 2. The particle sorting module according to clause 1, wherein the particle sorting module comprises:

[0167] a housing comprising a proximal end, a distal end, and a wall therebetween, wherein the opening is located in the wall; and

[0168] Droplet deflector.

[0169] 3. A particle sorting module according to clause 2, wherein the opening is located downstream of the droplet deflector.

[0170] 4. A particle sorting module according to any one of clauses 1 to 3, wherein one or more ports at the distal end of the housing are configured to couple to one or more sample collection containers.

[0171] 5. A particle sorting module according to clause 4, wherein the opening is configured for aligning the fluid flow with the one or more ports.

[0172] 6. A particle sorting module according to clause 4, wherein the opening is located between the droplet deflector and a distal end of the housing.

[0173] 7. A particle sorting module according to clause 6, wherein the opening is located at a position from 0.5 cm to 5 cm from the distal end of the housing.

[0174] 8. A particle sorting module according to clause 6, wherein the opening is located from 0.5 cm to 5 cm from the droplet deflector.

[0175] 9. A particle sorting module according to any one of clauses 1 to 8, wherein the opening comprises an optical adjustment component.

[0176] 10. A particle sorting module according to clause 9, wherein the optical conditioning comprises a focusing lens configured to focus optical irradiation on droplets from the deflected fluid flow.

[0177] 11. A particle sorting module according to any one of clauses 1 to 10, wherein the optical adjustment component comprises a transparent optical window.

[0178] 12. A particle sorting module according to clause 11, wherein the optical window comprises one or more reference identifiers.

[0179] 13. A particle sorting module according to clause 12, wherein the reference identifier comprises markings, each marking being associated with a boundary of one or more ports at the distal end of the housing.

[0180] 14. A particle sorting module according to any one of clauses 4 to 13, further comprising a waste collection container and two sample collection containers coupled to the distal end of the housing.

[0181] 15. A particle sorting module according to clause 14, wherein the opening comprises an optical window having a reference identifier associated with a boundary of each of the waste collection container and two sample collection containers.

[0182] 16. A particle sorting module according to any one of clauses 13 to 15, wherein the reference identifier comprises a marking indicating the centre of the distal end of the housing.

[0183] 17. A particle sorting module according to any one of clauses 1 to 16, further comprising:

[0184] a fluid pool nozzle at a proximal end of the housing, the fluid pool nozzle comprising an orifice; and

[0185] A sample interrogation zone is in fluid communication with the fluid reservoir nozzle orifice.

[0186] 18. A particle sorting module according to clause 17, further comprising a test tube located in the sample interrogation area.

[0187] 19. A particle sorting module according to clause 18, wherein the test tube is co-moulded with the housing.

[0188] 20. A particle sorting module according to any one of clauses 18 to 19, wherein the test tube comprises glass.

[0189] 21. A particle sorting module according to any one of clauses 18 to 19, wherein the test tube comprises plastic.

[0190] 22. A particle sorting module according to any one of clauses 1 to 21, further comprising a sample inlet at the proximal end of the housing.

[0191] 23. A particle sorting module according to any one of clauses 1 to 22, further comprising a sheath fluid inlet at the proximal end of the housing.

[0192] 24. A system comprising:

[0193] a particle sorting module comprising an opening configured to visualize droplets of the deflected fluid stream; and

[0194] An imaging sensor is configured to capture one or more images through the opening.

[0195] 25. The system of clause 24, further comprising a laser for irradiating the fluid flow through the opening.

[0196] 26. A system according to any one of clauses 24 to 25, wherein the particle sorting module comprises:

[0197] a housing comprising a proximal end, a distal end, and a wall therebetween, wherein the opening is located in the wall of the housing; and

[0198] Droplet deflector.

[0199] 27. The system of clause 26, further comprising one or more ports at a distal end of the housing, the one or more ports configured to couple to one or more sample collection containers.

[0200] 28. The system of clause 27, further comprising a processor, the processor comprising a memory operatively coupled to the processor, wherein the memory comprises instructions stored thereon that, when executed by the processor, cause the processor to:

[0201] capturing an image of an opening of the particle sorting module to obtain and image the opening; and

[0202] The pixel position of the boundary of each port at the distal end of the housing along the horizontal axis in the imaging opening is calculated.

[0203] 29. The system of clause 28, wherein calculating the pixel position comprises:

[0204] calculating a total number of pixels passing through the imaging opening along the horizontal axis; and

[0205] The total number of pixels is multiplied by a predetermined ratio.

[0206] 30. The system of clause 6, wherein the predetermined ratio is calculated by dividing the number of pixels in the imaging opening of each port boundary by the total number of pixels passing through the imaging opening along the horizontal axis.

[0207] 31. A system according to any of clauses 27 to 30, wherein the memory includes instructions stored thereon that, when executed by the processor, cause the processor to align the fluid flow with one of the ports at the distal end of the housing.

[0208] 32. The system of clause 31, wherein the memory includes instructions stored thereon that, when executed by the processor, cause the processor to align the fluid flow with a center of one of the ports at the distal end of the housing.

[0209] 33. A system according to any of clauses 30 to 32, wherein the memory includes instructions stored thereon that, when executed by the processor, cause the processor to align the fluid flow with the center of the distal end of the housing.

[0210] 34. The system of any of clauses 27 to 33, wherein the opening is located between the droplet deflector and a distal end of the housing.

[0211] 35. The system of clause 34, wherein the opening is located from 0.5 cm to 5 cm from the distal end of the housing.

[0212] 36. The system of clause 34, wherein the opening is located from 0.5 cm to 5 cm from the droplet deflector.

[0213] 37. A system according to any of clauses 24 to 36, wherein the opening includes an optical adjustment component.

[0214] 38. The system of clause 37, wherein the optical conditioning assembly comprises a focusing lens configured to focus optical radiation on droplets from the deflected fluid stream.

[0215] 39. A system according to any one of clauses 24 to 38, wherein the optical adjustment component includes a transparent optical window.

[0216] 40. The system of clause 39, wherein the optical window comprises one or more reference identifiers.

[0217] 41. The system of clause 40, wherein the reference identifier comprises markers, each marker being associated with the droplet deflection boundary.

[0218] 42. The system of any one of clauses 24 to 41, wherein the particle sorting module further comprises a waste collection container and two sample collection containers coupled to a distal end of the housing.

[0219] 43. A system according to clause 42, wherein the opening includes an optical window having a reference identifier associated with the deflection boundary of each of the waste collection container and the two sample collection containers.

[0220] 44. A system according to any of clauses 40 to 43, wherein the reference identifier includes a marking indicating the center of the distal end of the shell.

[0221] 45. The system according to any one of clauses 24 to 44, wherein the particle sorting module further comprises:

[0222] a fluid pool nozzle at a proximal end of the housing, the fluid pool nozzle comprising an orifice; and

[0223] A sample interrogation zone is in fluid communication with the fluid reservoir nozzle orifice.

[0224] 46. ​​The system of clause 45, wherein the particle sorting module further comprises a test tube located in the sample interrogation area.

[0225] 47. A system according to clause 46, wherein the test tube is co-molded with the housing.

[0226] 48. The system of any one of clauses 46 to 47, wherein the test tube comprises glass.

[0227] 49. The system of any one of clauses 46 to 47, wherein the test tube comprises plastic.

[0228] 50. The system of any one of clauses 24 to 49, further comprising a sample delivery subsystem in fluid communication with an inlet at a proximal end of the housing of the particle sorting module.

[0229] 51. The system of any one of clauses 24 to 50, further comprising a sheath fluid delivery subsystem in fluid communication with an inlet at a proximal end of the housing of the particle sorting module.

[0230] 52. The system of any one of clauses 24 to 51, further comprising a waste bin coupled to an outlet fluid of the particle sorting module.

[0231] 53. A method of aligning a fluid flow with one or more ports at a distal end of a particle sorting module, the method comprising:

[0232] capturing an image through an opening in the particle sorting module, the opening being configured to visualize droplets of the fluid stream;

[0233] calculating a pixel position of said boundary of each said port at said distal end in an imaging opening along a horizontal axis; and

[0234] The fluid stream is aligned with each port based on the calculated pixel location of the boundary of the port in the imaging opening.

[0235] 54. The method of clause 53, wherein calculating the pixel position comprises:

[0236] calculating a total number of pixels passing through the imaging opening along the horizontal axis; and

[0237] The total number of pixels is multiplied by a predetermined ratio.

[0238] 55. The method of clause 54, further comprising calculating the predetermined ratio by dividing the number of pixels in the imaging opening of each port boundary by the total number of pixels passing through the imaging opening along the horizontal axis.

[0239] 56. A method according to any one of clauses 53 to 55, wherein the particle sorting module comprises a waste collection container and two sample collection containers.

[0240] 57. The method of any of clauses 53 to 56, further comprising irradiating the fluid flow through the opening with a laser.

[0241] 58. A method according to any one of clauses 53 to 57, wherein the particle sorting module comprises:

[0242] a closed housing comprising a proximal end, a distal end and a wall therebetween, wherein the opening is located in the wall of the housing; and

[0243] Droplet deflector.

[0244] 59. The method of clause 58, wherein the opening is located between the droplet deflector and a distal end of the housing.

[0245] 60. The method of clause 59, wherein the opening is located from 0.5 cm to 5 cm from the distal end of the housing.

[0246] 61. The method of clause 59, wherein the opening is located from 0.5 cm to 5 cm from the droplet deflector.

[0247] 62. A system according to any of clauses 53 to 61, wherein the opening includes an optical adjustment component.

[0248] 63. A method according to clause 62, wherein the optical adjustment component includes a focusing lens.

[0249] 64. A method according to clause 62, wherein the optical adjustment component includes a transparent optical window.

[0250] 65. A kit comprising:

[0251] a particle sorting module comprising an opening configured to visualize droplets of the deflected fluid stream; and

[0252] One or more sample collection containers, the one or more sample collection containers are used to be coupled to the particle sorting module.

[0253] 66. The kit according to clause 65, further comprising a droplet deflector.

[0254] 67. A kit according to any one of clauses 65 to 66, wherein the particle sorting module comprises a sample inlet and a sheath fluid inlet.

[0255] 68. The kit according to clause 67, further comprising one or more connectors for coupling the particle sorting module to a sample fluid delivery system and a sheath fluid delivery system.

[0256] 69. A kit according to any one of clauses 65 to 68, wherein one or more of the particle sorting module, sample collection container and connector are present in a container.

[0257] 70. A kit according to clause 69, wherein one or more of the particle sorting module, sample collection container and connector are present in a pocket.

[0258] 71. A kit according to any one of clauses 65 to 70, wherein the kit components are sterile.

[0259] In at least some of the previously described embodiments, one or more elements used in one embodiment may be used interchangeably in another embodiment, unless such replacement is technically infeasible. It will be clear to those skilled in the art that various other omissions, additions, and modifications may be made to the above methods and structures without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter defined by the appended claims.

[0260] It will be understood by those skilled in the art that, in general, the terms used herein (and particularly in the appended claims (e.g., the bodies of the appended claims)) are often used as "open" terms (e.g., the term "comprising" should be considered "including but not limited to", the term "having" should be considered "having at least one", the term "including" should be considered "including but not limited to", etc.). It will be further understood by those skilled in the art that if a specific number of an introduced claim is intended to be recited, such intent will be explicitly recited in the claim, and in the absence of such a recitation, such intent is absent. For example, to aid understanding, the following appended claims may contain the use of the preamble phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed as implying that a claim recitation introduced by the indefinite article "a" or "an" will limit any specific claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the preamble phrases "one or more" or "at least one" and an indefinite article such as "a" or "an" (e.g., "a" and / or "an" should typically be interpreted as meaning "at least one" or "one or more"); the same applies to the use of definite articles used to introduce claim recitations. Furthermore, even if a specific number of an introduced claim recitation is explicitly recited, one skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the simple recitation "two recitations," without other modifications, means at least two recitations, or two or more recitations). Additionally, in those cases where the conventional phrase "at least one of A, B, and C, etc." is used, such phrase is generally constructed in a sense that one skilled in the art should understand the convention (e.g., "a system having at least one of A, B, and C" would include, but is not limited to, a system having only A, a system having only B, a system having only C, a system having A and B, a system having A and C, a system having B and C, and / or a system having A, B, and C, etc.). In those cases where the conventional phrase "at least one of A, B, or C, etc." is used, such phrase is generally constructed in a sense that one skilled in the art should understand the convention (e.g., "a system having at least one of A, B, or C" would include, but is not limited to, a system having only A, a system having only B, a system having only C, a system having A and B, a system having A and C, a system having B and C, and / or a system having A, B, and C, etc.). Those skilled in the art will further understand that transitional words or phrases indicating two or more alternative terms, whether in the specification, claims or drawings, should be understood to include the possibility of one, any, or both of the terms. For example, the phrase "A or B" will be understood to include the possibility of "A" or "B" or "A and B".

[0261] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0262] As will be understood by those skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also include any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily considered to fully describe and allow the same range to be decomposed into at least equal halves, one-third, one-quarter, one-fifth, one-tenth, etc. As a non-limiting example, each range discussed herein can be easily decomposed into a lower third, a middle third, and an upper third, etc. As will be understood by those skilled in the art, all languages ​​(such as "at most", "at least", "greater than", "less than", etc.) include the cited numbers and refer to the ranges that can be subsequently subdivided into sub-ranges as described above. Finally, as will be understood by those skilled in the art, the range includes each individual member. Therefore, for example, a group with 1-3 articles refers to a group with 1, 2 or 3 articles. Similarly, a group with 1-5 articles refers to a group with 1, 2, 3, 4 or 5 articles, and the like.

[0263] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to one skilled in the art that certain changes and modifications may be made thereto in accordance with the teachings of the invention without departing from the spirit or scope of the appended claims.

[0264] Therefore, the above only illustrates the principle of the present invention.It should be clear that those skilled in the art will be able to design various arrangements, although not explicitly described or shown here, these arrangements embody the principle of the present invention, and are included in the spirit and scope of the present invention.In addition, all examples and conditional language described herein are mainly intended to help readers understand the concept of the principle of the present invention and the inventor's further development contribution to this field, and should be interpreted as not being limited to these specifically described examples and conditions.In addition, all statements describing the principles, aspects and embodiments of the present invention and its specific examples herein are intended to include their structural and functional equivalents.In addition, it is expected that such equivalents include currently known equivalents and equivalents developed in the future, that is, any element of the performance of the same function developed, regardless of its structure.In addition, anything disclosed herein is not intended to be used exclusively for the public, regardless of whether the disclosure is clearly described in the claims.

[0265] Therefore, the scope of the present invention is not limited to the exemplary embodiments shown and described herein. Instead, the scope and spirit of the present invention are embodied by the appended claims. In the claims, 35 USC § 112 (f) or 35 USC § 112 (6) is expressly defined as being invoked for limitation in the claims only when the exact phrase "means" or the exact phrase "step" is recited at the beginning of such limitation in the claims; if the exact phrase is not used in the limitation in the claims, 35 USC § 112 (f) or 35 USC § 112 (6) will not be invoked.

Claims

1. A particle sorting module, the particle sorting module comprising: A housing, the housing comprising a proximal end, a distal end and a wall therebetween, the housing further comprising: Droplet deflectors; one or more sample collection ports at a distal end of the housing; as well as An opening located in the wall between the droplet deflector and a distal end of the housing and configured to visualize droplets of the deflected fluid stream, wherein the opening includes an optical window having one or more reference identifiers configured to align the fluid stream with one or more sample collection ports. 2 . The particle sorting module of claim 1 , wherein the one or more ports at the distal end of the housing are configured to be coupled to one or more sample collection containers.

3. The particle sorting module according to claim 1, wherein: The one or more reference identifiers include one or more markings associated with a boundary of each of the one or more ports at the distal end of the housing.

4. The particle sorting module according to claim 1, wherein: The opening includes an optical adjustment component.

5. The particle sorting module according to claim 4, wherein: The optical conditioning assembly includes a focusing lens configured to focus optical radiation on droplets from the deflected fluid stream.

6. The particle sorting module of claim 2, further comprising a waste collection container and two sample collection containers coupled to a distal end of the housing.

7. The particle sorting module according to claim 6, wherein: The one or more reference identifiers include markings associated with boundaries of each of the waste collection container and two sample collection containers.

8. The particle sorting module according to any one of claims 1 to 7, further comprising: a fluid pool nozzle at a proximal end of the housing, the fluid pool nozzle comprising an orifice; and A sample interrogation zone is in fluid communication with the orifice of the fluid reservoir nozzle.

9. The particle sorting module of claim 8, further comprising a test tube located in the sample interrogation area.

10. The particle sorting module according to any one of claims 1 to 7, further comprising a sample inlet at the proximal end of the housing and a sheath fluid inlet at the proximal end of the housing.

11. A system comprising: A particle sorting module according to any one of claims 1 to 10.

12. A method of aligning a fluid flow with one or more ports at a distal end of a particle sorting module, the method comprising: capturing an image through an opening in a particle sorting module according to any one of claims 1 to 10; calculating a pixel position of a boundary of each port at the distal end in the imaging opening along a horizontal axis; as well as The fluid stream is aligned with each port based on the calculated pixel location of the boundary of the port in the imaging opening.

13. A kit comprising: A particle sorting module according to any one of claims 1 to 10.

Images