NL2036035B1

NL2036035B1 - Cartridge for fluid with particles

Info

Publication number: NL2036035B1
Application number: NL2036035A
Authority: NL
Inventors: Chou Hou-Pu; Wang Cheng; Robles Michael; L Luengo Hendriks Christian
Original assignee: Deepcell Inc
Priority date: 2023-09-21
Filing date: 2023-10-13
Publication date: 2025-03-26

Abstract

An apparatus includes a first ﬂuidic path to receive a ﬂuid containing particles, a first sorting region, and a second sorting region. The first sorting region includes a first junction joining the first ﬂuidic path with second and third ﬂuidic paths; and a set of valves to selectively permit or prevent communication of ﬂuid along the second or third ﬂuidic paths. The second sorting region includes a second junction joining the second ﬂuidic path With fourth and fifth ﬂuidic paths; and another set of valves to selectively permit or prevent communication of ﬂuid along the fourth or ﬁfth ﬂuidic paths. An outlet port is to extract ﬂuid from the third ﬂuidic path. A first well is to receive ﬂuid from the fourth ﬂuidic path. A second well is to receive ﬂuid from the fifth ﬂuidic path.

Description

CARTRIDGE FOR FLUID WITH PARTICLES

BACKGROUND

[0091] The properties of cells may be analyzed to diagnose diseases and other conditions. Such analysis may include evaluation of cell morphology to determine cell type (e.g., stem cell or differentiated cell) or cell state (e.g., healthy state or disease state). In some cases, cells may be directed through a channel of a cartridge, under fluidic guidance, through a microscope imaging field. The images captured through the microscope may be processed to evaluate cell morphology. While a variety of devices, systems, and methods have been made and used to process and analyze cells, it is believed that no one prior to the inventor(s) has made or used the devices and techniques described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[0002] FIG. 1 depicts a schematic view of an example of a cell analysis system.

[6003] FIG. 2 depicts a perspective view of an example of a cartridge that may be used in examples of the cell analysis system of FIG. 1.

[0604] FIG. 3 depicts an exploded perspective view of the cartridge of FIG. 2.

[6005] FIG. 4 depicts a perspective view of an example underside of a first layer of the cartridge of

FIG. 2.

[6006] FIG. 5 depicts a perspective view of an example underside of a second layer of the cartridge of

FIG. 2.

[0007] FIG. 6 depicts a top plan view of the cartridge of FIG. 2.

[9608] FIG. 7 depicts an enlarged view of an example fluid input region of the cartridge of FIG. 2.

[0009] FIG. 8 depicts an enlarged perspective view of the underside of the second layer of FIG. 5, showing part of a sample fluid receiving region of the fluid input region of FIG. 7.

[8010] FIG. 9 depicts an enlarged view of an example mixing structure of the fluid input region of

FIG. 7.

[0011] FIG. 10 depicts a cross-sectional view of the mixing structure of FIG. 9, taken along line 9-9 of FIG. 9.

[6012] FIG. 11 depicts a perspective schematic view of a portion of the mixing structure of FIG. 9.

[0013] FIG. 12A depicts a schematic view of fluid flow dynamics in a portion of the fluid input region of FIG. 7 just upstream of the mixing structure of FIG. 9.

[6014] FIG. 12B depicts a schematic view of fluid flow dynamics in a portion of the fluid input region of FIG. 7 just downstream of the mixing structure of FIG. 9.

[0015] FIG. 13 depicts an enlarged top plan view of an example of a primary sorting region of the cartridge of FIG. 2.

[0016] FIG. 14A depicts a cross-sectional view of the primary sorting region of FIG. 13, taken along line 14-14 of FIG. 13, with an example of a first valve in an open state and an example of a second valve in an open state.

[0017] FIG. 14B depicts a cross-sectional view of the primary sorting region of FIG. 13, taken along line 14-14 of FIG. 13, with the second valve in a closed state and the first valve in the open state.

[0018] FIG. 14C depicts a cross-sectional view of the primary sorting region of FIG. 13, taken along line 14-14 of FIG. 13, with the second valve in the open state and the first valve in a closed state.

[6019] FIG. 15 depicts an enlarged top plan view of an example of a secondary sorting region of the cartridge of FIG. 2.

[8020] FIG. 16 depicts an enlarged top plan view of a region of the cartridge of FIG. 2 including an example set of wells downstream of the secondary sorting region of FIG. 15.

[0921] FIG. 17 depicts an enlarged top plan view of a region of the cartridge of FIG. 2 including examples of venting features.

[0022] FIG. 18 depicts an enlarged perspective view of the underside of the first layer of FIG. 4, showing the venting features of FIG. 17.

[0023] FIG. 19 depicts an enlarged top plan view of a region of another example of a cartridge that may be used in examples of the cell analysis system of FIG. 1, with examples of other venting features.

[6024] FIG. 20A depicts enlarged top plan view of a region of the cartridge of FIG. 2 including example flush ports and example fluid input ports, with a schematic representation of a set of fluid conduits coupled with the fluid input ports.

[0025] FIG. 20B depicts an enlarged top plan view of the region of FIG. 20A, with a schematic representation of a set of fluid conduits coupled with the flush ports.

[0026] FIG. 21A depicts enlarged top plan view of another region of the cartridge of FIG. 2 including other example flush ports, an example fluid input port, and an example fluid output port, with a schematic representation of a set of fluid conduits coupled with the fluid input port and the fluid output port.

[6027] FIG. 21B depicts an enlarged top plan view of the region of FIG. 20A, with a schematic representation of a set of fluid conduits coupled with the flush ports.

[0028] FIG. 22 depicts enlarged top plan view of a region of the cartridge of FIG. 2 including an example optical fiducial region.

[0029] FIG. 23 depicts an enlarged top plan view of a first corner of the optical fiducial region of FIG. 22.

[0030] FIG. 24 depicts an enlarged top plan view of a second corner of the optical fiducial region of

FIG. 22.

[0031] FIG. 25 depicts an enlarged top plan view of a third corner of the optical fiducial region of FIG. 22.

[0032] FIG. 26 depicts an enlarged top plan view of a fourth corner of the optical fiducial region of

FIG. 22.

[0033] FIG. 27 depicts an enlarged cross-sectional view of the cartridge of FIG. 2, taken along line 27-27 of FIG. 22, showing example focusing features.

[0034] FIG. 28 depicts a schematic representation of particles flowing along a fluidic channel of the cartridge of FIG. 2, with the particles separated in an example of two vertical regions.

[0035] FIG. 29 depicts an enlarged top plan view of an example sorting region of another example of a cartridge that may be used in examples of the cell analysis system of FIG. 1.

[0036] FIG. 30A depicts a cross-sectional view of a valve of the sorting region of FIG. 29, taken along line 30-30 of FIG. 29, with the valve in an open state.

[6037] FIG. 30B depicts a cross-sectional view of a valve of the sorting region of FIG. 29, taken along line 30-30 of FIG. 29, with the valve in a closed state.

[0038] FIG. 31 depicts a schematic view of another example of a cartridge that may be used in examples of the cell analysis system of FIG. 1.

DETAILED DESCRIPTION

[6039] The following detailed description of certain examples will be better understood when read in conjunction with the appended drawings. To the extent that the figures illustrate diagrams of the functional blocks of various examples, the functional blocks are not necessarily indicative of the division between hardware components. Thus, for example, one or more of the functional blocks (e.g., processors or memories) may be implemented in a single piece of hardware (e.g., a general purpose signal processor or random access memory, hard disk, or the like). Similarly, the programs may be stand-alone programs, may be incorporated as subroutines in an operating system, may be functions in an installed software package, and the like. It should be understood that the various examples are not limited to the arrangements and instrumentality shown in the drawings.

[0040] L Terminology

[0041] Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising” means various components may be co-jointly employed in the methods and articles (e.g., compositions and apparatuses including device and methods). For example, the term “comprising” will be understood to imply the inclusion of any stated elements or steps but not the exclusion of any other elements or steps. In general, any of the apparatuses and methods described herein should be understood to be inclusive, but all or a sub-set of the components and/or steps may alternatively be exclusive and may be expressed as “consisting of” or alternatively “consisting essentially of” the various components, steps, sub-components, or sub-steps. Furthermore, references to “one example” are not intended to be interpreted as excluding the existence of additional examples that also incorporate the recited features. The use of “including,” “comprising,” “having,” or “in which,” and variations thereof, herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items.

[0042] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items and may be 5 abbreviated as “/”.

[0043] When used in the claims, the term “set” should be understood as one or more things which are grouped together. Similarly, when used in the claims “based on” should be understood as indicating that one thing is determined at least in part by what it is specified as being “based on.” Where one thing is required to be exclusively determined by another thing, then that thing will be referred to as being “exclusively based on” that which it is determined by.

[0044] Spatially relative terms, such as “under,” “below,” “lower,” “over,” “upper,” and the like, may be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is inverted, elements described as “under” or “beneath” other elements or features would then be oriented “over” the other elements or features. Thus, the term “under” may encompass both an orientation of over and under. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly. Similarly, the terms “upwardly,” “downwardly,” “vertical,” “horizontal,” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise. In addition, terms such as “outer” and “inner” are used herein for purposes of description and are not intended to indicate or imply relative importance or significance.

[0045] When a feature or element is herein referred to as being “on” or “over” another feature or element, it may be directly on or over the other feature or element; or intervening features and/or elements may also be present. In other words, when a feature or element is herein referred to as being “on” or “over” another feature or element, it may be indirectly on or over the other feature or element. In contrast, when a feature or element is referred to as being “directly on” or “directly over” another feature or element, there are no intervening features or elements present.

[0946] When a feature or element is referred to as being “mounted,” “connected.” “supported,” “attached,” or “coupled” to another feature or element, it may be directly mounted, connected,

supported, attached, or coupled to the other feature or element or intervening features or elements may be present. In contrast, when a feature or element is referred to as being “directly mounted,” “directly connected,” “directly supported,” “directly attached,” or “directly coupled” to another feature or element, there are no intervening features or elements present.

Although described or shown with respect to one embodiment, the features and elements so described or shown may apply to other embodiments. It will also be appreciated by those skilled in the art that references to a structure or feature that is disposed “adjacent” another feature may have portions that overlap or underlie the adjacent feature.

[0047] As used herein in the specification and claims, including as used in the examples and unless otherwise expressly specified, the terms “about” or “approximately” for any numerical values or ranges indicate a suitable dimensional tolerance, or other form of reasonable expected range, that allows the part or collection of components to function for its intended purpose as described herein. More specifically, “about” or “approximately” may refer to the range of values that are within £10% of the recited value (e.g., “about 100” may refer to the range of values from 90 to 110, including 90, 110, 100, and all other values within the range of 90 and 110). Any numerical values given herein should also be understood to include about or approximately that value unless the context indicates otherwise. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Any numerical range recited herein is intended to include all sub- ranges subsumed therein. The terms “approximately” and “about” are thus utilized herein to represent the degree by which a quantitative representation may vary from a stated reference without resulting in a change in the basic function of the subject matter at issue.

[0048] The term “substantially” is also utilized herein to represent the degree by which a quantitative representation may vary from a stated reference without resulting in a change in the basic function of the subject matter at issue. The term “substantially” shall therefore be understood to include a range of conditions or results that provide a functional equivalent to an explicitly stated condition or result. For instance, if a task is “substantially complete,” the result of the task having been substantially completed is functionally equivalent to the result that would have been achieved if the task had been perfectly completed. As another non-limiting example, a component that is “substantially straight” or “substantially flat,” an apparatus including a component that is “substantially straight” or “substantially flat” may provide a result or effect that is functionally equivalent to a result or effect that would be achieved by the same apparatus including the same component in a pertectly straight or perfectly flat configuration. The range implied by the term “substantially” should also be read to include the perfect result that is within that range. Thus, the term “substantially complete” shall be read as including “perfectly complete” while also including a range of completeness that is functionally equivalent to perfectly complete. As another example, terms such as “substantially straight” and “substantially flat” shall be read as including “perfectly straight” and “perfectly flat,” respectively; while also including a range of straightness or flatness that is functionally equivalent to perfectly straight or flat, respectively. As with the terms “approximately” and “about,” the term “substantially” may indicate a suitable dimensional tolerance, or other form of reasonable expected range, that allows a part or collection of components to function for its intended purpose as described herein.

[0049] The term “perpendicular” shall be understood to include arrangements where one element (e.g, surface, feature, component, axis, etc.) defines an angle of 90 degrees with another element {e.g., surface, feature, component, axis, etc.). The term “perpendicular” shall also be understood to include arrangements where one element (e.g., surface, feature, component, axis, etc.) defines an angle of approximately 90 degrees with another element (e.g., surface, feature, component, axis, etc).

[6050] It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value,” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “X” is disclosed the “less than or equal to X” as well as “greater than or equal to X” (e.g., where X is a numerical value) is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0051] Although the terms “first” and “second” may be used herein to describe various features/elements (including steps), these features/elements should not be limited by these terms, unless the context indicates otherwise. These terms are used to distinguish one feature/element from another feature/element, and unless specifically pointed out, do not denote a certain order. Thus, a first feature/element discussed below could be termed a second feature/element, and similarly, a second feature/element discussed below could be termed a first feature/element without departing from the teachings of the present invention. The terms “first,” “second,” and “third,” etc. are thus used merely as labels, and are not intended to impose numerical requirements on their objects.

[0052] As used herein, the terms “system,” “apparatus,” and “device” may be read as being interchangeable with each other. A system, apparatus, and device may each include a plurality of components having various kinds of structural and/or functional relationships with each other.

[6053] The term “fluid” shall be understood to include liquids and gases, including pneumatic pressure. Similarly, “fluidic communication” shall be understood to include the communication of liquids and the communication of gases, including pneumatic pressure.

[06054] The term “morphology” or “morphological characteristic” of a cell as used herein generally refers to the form, structure, and/or configuration of the cell. The morphology of a cell may comprise one or more aspects of a cell's appearance, such as, for example, shape, size, arrangement, form, structure, pattern(s) of one or more internal and/or external parts of the cell, or shade (e.g., color, greyscale, etc.). Non-limiting examples of a shape of a cell may include, but are not limited to, circular, elliptic, dumbbell, star-like, flat, scale-like, columnar, invaginated, having one or more concavely formed walls, having one or more convexly formed walls, prolongated, having appendices, having cilia, having angle(s), having corner(s), etc. A morphological feature of a cell may be visible with treatment of a cell (e.g., small molecule or antibody staining). In other examples, the morphological feature of the cell may not and need not require any treatment to be visualized in an image or video.

[0055] The terms “unstructured” or “unsorted,” as used interchangeably herein, generally refers to a mixture of cells {e.g., an initial mixture of cells) that is not substantially sorted (or rearranged) into separate partitions. An unstructured population of cells may comprise at least two types of cells that can be distinguished by exhibiting different properties (e.g., one or more physical properties, such as one or more different morphological characteristics as disclosed herein).

The unstructured population of cells may be a random (or randomized) mixture of the at least two types of cells. The cells as disclosed herein may be viable cells. A viable cell, as disclosed herein, may be a cell that is not undergoing necrosis or a cell that is not in an early or late apoptotic state. In other examples, the cells may not and need not be viable (e.g., fixed cells).

[0056] The term “resilient” as used herein refers to a material property where the material has shape memory and stiffness such that it is structurally biased toward a neutral shape or structural arrangement. As an example, a resilient member may have a resilient bias toward a neutral shape or structural arrangement where the resilient member is straight along a central longitudinal axis. That same resilient member may be deformed relative to the neutral shape or structural arrangement, such as by being bent away from that central longitudinal axis, in response to a force (e.g., when a force is imparted on the resilient member, where the force has a directional component (hat is transverse to the central longitudinal axis). While the resilient member is being deformed relative to the neutral shape in response to the force, the resilient member may be under stress whereby the resilient property of the material of the resilient member generates a force in a direction that is opposite to the force that is causing the deformation of the resilient member. In other words, the resilient property of the material of the resilient member may impart a mechanical bias urging the resilient member back toward the neutral shape or structural arrangement. After the force causing the deformation of the resilient member is removed, the resilient bias of the material of the resilient member may cause the resilient member to return to (or at least toward) the neutral shape or structural arrangement. While the foregoing example provides a straight configuration as a neutral shape or structural arrangement, other examples of resilient members may have other kinds of neutral shapes or structural arrangements.

[0057] IL Example of Cell Analysis System

[0058] The systems and methods described herein may be utilized to analyze a cell and/or sort (or partition) the cell from a population of cells. A cell may be directed through a flow channel, and one or more imaging devices (e.g., sensor(s), camera(s)) may capture one or more images/videos of the cell passing through the flow channel. Subsequently, the image(s)/video(s) of the cell may be analyzed in real-time, such that a decision may be made in real-time (e.g., automatically by the machine learning algorithm) to determine (i) whether to sort the cell or not and/or (ii) which sub-channel of a plurality of sub-channels to sort the cell into.

[0059] FIG. 1 shows an example of a cell analysis system (100), which may be used to capture images of cells, and apply machine learning or artificial intelligence to analyze the captured images of the cells, and automatically sort the cells based on the analysis. System (100) of this example includes a pump (110) that is operable to drive a sample cell-containing fluid from a reservoir (112) into a cartridge (120). Cartridge (120) may be provided as a modular component, such that cartridge (120) may be readily replaced within system (100) (e.g., to analyze different batches of sample cells, etc.). The remaining components of system (100) that do not get replaced each time cartridge (120) is replaced may be collectively referred to as “the instrument.” The instrument of system (100) may include pump (110); or pump (110) may be considered as a separate component such that a different pump (110) may be used when a different batch of sample cells is being analyzed. Similarly, the instrument of system (100) may include reservoir (112); or reservoir (112) may be considered as a separate component such that a different reservoir (112) may be used when a different batch of sample cells is being analyzed.

[0660] In some examples, reservoir (112) comprises a syringe barrel; and pump (110) comprises a syringe pump. In other examples, pump (110) may take any other suitable form, including but not limited to a gravity feed, a peristaltic pump, etc. Reservoir (112) may also take any other suitable form, including but not limited to a vial, tube, etc. The sample in reservoir (112) may be prepared by fixation and staining; and may contain viable cells. The fluid in which the sample cells are contained may include an aqueous solution (e.g., water, buffer, saline, etc.), an oil, or any other suitable fluid.

[0061] Cartridge (120) includes a flow channel (122) fluidically coupled with pump (110), such that pump (110) is operable to drive the sample cell-containing fluid from reservoir (112) through flow channel (122). Cartridge (120) may comprise a microfluidic chip, a flow cell, or any other kind of structure through which fluid may flow; and through which cells in the fluid may be imaged. A light source (130) generates light for such imaging. In particular, an optical assembly (132) directs light from light source (130) toward an imaging region of flow channel (122). In some examples, light source (130) comprises a source of incoherent white light, such as an arc lamp, etc. In other examples, light source (130) may take any other suitable form.

Optical assembly (132) may comprise any suitable number and/or arrangement of lenses and/or other elements as will be apparent to those skilled in the art in view of the teachings herein.

[0082] The light from light source (130), as directed by optical assembly (132), illuminates cells as the cells pass through the imaging region of flow channel (122). An objective lens assembly (140) is positioned on the opposite side of the imaging region of flow channel (122), magnifies the images of cells passing through the imaging region of flow channel (122), and directs the magnified images to a camera (142). Objective lens assembly (140) and camera (142) thus cooperate to capture high resolution images of cells that pass through the imaging region of flow channel (122), as illuminated by light source (130) and optical assembly (132). By way of example only, objective lens assembly (140) may provide magnification ranging from approximately 10x to approximately 200%. In other examples, objective lens assembly (140)

may provide any other suitable level of magnification. By way of further example only, camera (142) may provide an exposure time ranging from approximately 0.001 ys to approximately 1 ms. In other examples, camera (142) may provide any other suitable exposure time. In examples described below, objective lens assembly (140) and camera (142) have an optical axis along the z-dimension.

[0063] An image processing module (144) receives images from camera (142) and processes those received images in real time. Image processing module (144) may include one or more processors, one or more memories, and various other suitable electrical components. Image processing module (144) may also include software, firmware and/or hardware. In some examples, image processing module (144) is in communication with a remote server and/or with other components. The one or more processors of image processing module (144) may comprise one or more of a programmable processor, a programmable controller, a microprocessor, a microcontroller, a graphics processing unit (GPU), a digital signal processor (DSP), a reduced-instruction set computer (RISC), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA), a field programmable logic device (FPLD), a logic circuit, and/or another logic-based device executing various functions including the ones described herein.

[0064] Image processing module (144) may utilize any of a number of techniques to classify or otherwise analyze images of cells captured by camera (142). For instance, cell image data may be analyzed to plot a plurality of cells into a cell clustering map. The image data may comprise tag-free images of single cells. In other examples, the image data may comprise images of single cells that are tagged (e.g., with a heterologous marker). The image data may be processed to generate a cell morphology map. The cell morphology map may comprise a plurality of morphologically distinct clusters corresponding to different types or states of the cells. In some cases, a classifier (e.g., a cell clustering machine learning algorithm or deep learning algorithm) may be trained by using the cell morphology map. The classifier may be configured to classify a cell image sample based on its proximity, correlation, or commonality with one or more of the morphologically distinct clusters. Thus, in some cases, the classifier may be used to classify the sample cell image sample accordingly. This classification may be fully automatic, such that the classification is accomplished solely by software executed through image processing module (144), without additional human operator review of the sample cell image. In some other examples, the classification is at least partially manual such that a human operator verifies or otherwise intervenes to inform or approve the classification of the sample cell image.

[6065] Regardless of the technique(s) used by image processing module (144) to classify or otherwise analyze images of cells captured by camera (142), system (100) may provide sorting of cells in cartridge (120) based on such image processing. In some examples, cartridge (120) may include two or more outlet channels from flow channel, and system (100) may automatically activate one or more valves to direct an imaged cell through a selected one of those outlet channels based on the image analysis of the cell by image processing module (144). For instance, a certain outlet channel may be selected if classification or other analysis by image processing module (144) determines that the cell appears to be a certain cell type of interest.

In other examples, system (100) may provide cell sorting in any other suitable fashion; and based on any suitable criterion or criteria. In some other examples, system (100) provides imaging and analysis of cells without subsequent sorting of cells. In such examples, the imaged cells may remain contained in cartridge (120) after imaging or may exit cartridge (120) via an outlet port after imaging.

[0066] As noted above, the remaining components of system (100) that do not get replaced each time cartridge (120) is replaced may be collectively referred to as “the instrument.” In some examples, the instrument includes light source (130), optical assembly (132), objective lens assembly (140), camera (142), and various other components that removably receive cartridge (120) and provide any fluidic couplings, etc., that are needed for system (100) to perform properly. The instrument may further include image processing module (144). In other examples, image processing module (144) may be provided as a separate component (e.g., computer, etc.) that is coupled with camera (142) of the instrument to process images captured by camera (142). As further noted above, the instrument may further include or omit either or both of pump (110) and/or reservoir (112).

[0067] TI. Example of Cartridge

[0068] A. Overview

[6069] FIGS. 2-6 show an example of a form that may be taken by cartridge (120). In particular, FIGS. 2-6 show an example of a cartridge (200) that may be used in system (100) to provide imaging, analysis, and sorting of cells flowed through cartridge (200). Cartridge (200) of this example includes a first layer (300), a second layer (400), and a third layer (500). In the present example each of layers (300, 400) comprises a polymer (e.g., a siloxane-containing polymer, such as polydimethylsiloxane (PDMS), thermoset plastic, hydrogel, thermoplastic elastomer (TPE),

thermoplastic polyurethane (TPU), polycarbonate (PC), polystyrene (PS), poly(methyl methacrylate) (PMMA), cyclic olefin copolymer (COC), etc.). Layer (500) may comprise a glass (e.g., borosilicate or other silicate glass, etc.) or a polymer (e.g., polycarbonate (PC), polystyrene (PS), poly(methyl methacrylate) (PMMA), cyclic olefin copolymer (COC), etc.).

In other examples, each layer (300, 400, 500) may comprise any other suitable material or combination of materials, Layers (300, 400, 500) are arranged such that a top surface (302) of layer (300) is exposed, a bottom surface (304) of layer (300) is apposed with a top surface (402) of layer (400), and a bottom surface (404) of layer (400) is apposed with a top surface (502) of layer (500). Layer (500) acts as a substrate, providing structural support to layers (300, 400), with a bottom surface (502) of layer (500) being placed upon a mounting surface (not shown) in an instrument of system (100).

[0070] As shown in FIGS. 2-4, layer (300) is substantially thicker than layer (400). As also shown in

FIGS. 2-4, layer (300) includes a plurality of fluid input ports (310), a plurality of fluid output ports (312), plurality of pneumatic ports (320), and a plurality of well channels (340), and a pair of tabs (306). Ports (310, 312, 320) are in the form of channels formed through the entire thickness of layer (300), such that each port (310, 312, 320) is open at top surface (302) and at bottom surface (304). In some examples, fluid input conduits of system (100) are inserted into fluid input ports (310), tluid output conduits of system (100) are inserted into fluid output ports (312), and pneumatic input conduits of system (100) are inserted into pneumatic ports (320).

An instrument of system (100) may thus communicate liquids to cartridge (200) via fluid input ports (310), receive liquids from cartridge (200) via fluid output ports (312), and provide pneumatic pressure to cartridge (200) via pneumatic ports (320). In some examples, the conduits of system (100) that are coupled with ports (310, 312, 320) comprise flexible tubes.

In other examples, such conduits may take any other suitable form.

[0071] As shown in FIG. 4, bottom surface (304) of layer (300) includes a plurality of pneumatic channels (322), with each pneumatic channel (322) being in fluidic communication with a corresponding pneumatic port (320). Pneumatic channels (322) thus receive pneumatic pressure communicated through pneumatic ports (320). Such pneumatic pressure may be used to provide deflection of regions of layer (400) underlying pneumatic channels (322), such that those regions of layer (400) may be operated as pinch valves to close fluidic communication between layers (400, 500) as described in greater detail below. Each pneumatic channel (322) is formed as a recess in bottom surface (304), such that pneumatic pressure is communicated along the space defined collectively by top surface (402) of layer (400) and each pneumatic channel (322). In other words, top surface (402) of layer (400) defines a bottom of each pneumatic channel (322).

[0072] As shown in FIGS. 2-3 and 5, layer (400) includes a plurality of fluid input ports (410), a plurality of fluid output ports (412), and a plurality of well openings (440). Ports (410, 412) and well openings (440) are in the form of openings formed through the entire thickness of layer (400). Fluid input ports (410) of layer (400) are positioned to align with fluid input ports (310) of layer (300), such that fluid may be communicated from an instrument of system (100) through layer (400) via fluid input ports (310, 410). Fluid output ports (412) of layer (400) are positioned to align with fluid output ports (312) of layer (300). such that fluid may be communicated through layer (400) to an instrument of system (100) via fluid output ports (312, 412). Well openings (440) of layer (400) are positioned to align with well channels (340) of layer (300), such that fluid may be communicated through layer (400) via well openings (440) into well channels (340).

[0073] Well openings (440), well channels (340), and top surface (502) of layer (500) thus cooperate i5 to define a plurality of wells (750, 752, 754, 756, 758, 760), in which particles in fluid may be stored. In some examples, a removable layer (e.g., tape, film, foil, etc.) or other removable cover is provided over openings (440), such as to minimize, and in some instances even prevent, evaporation from the underlying wells, to minimize, and in some instances even prevent, contamination of the underlying wells, and/or for other reasons. In some such examples, an operator may remove such a layer or cover from a well to retrieve fluid from the well (e.g., via pipette, etc). It should be understood that layer (400) does not include pneumatic openings or ports in this example, such that pneumatic pressure is not communicated through layer (400).

[0074] As shown in FIG, 5, bottom surface (404) of layer (400) includes a fluidic test input region (450), a pair of fluidic flush input regions (460), a pair of fluidic flush output regions (462), a sample fluid receiving region (600), a flow control fluid receiving region (620), a flow drive fluid receiving region (670), and a sample output region (680). A plurality of fluidic channels (442, 452, 464, 604, 662. 664, 672) are formed as recesses in bottom surface (304), such that fluid is communicated along the space defined collectively by top surface (502) of layer (500) and each fluidic channel (442, 452, 464, 604, 662. 664, 672).

[0075] Fluidic channel (452) terminates within layer (400), such that fluidic channel (452) lacks any kind of fluidic outlet. In some scenarios, a fluid input conduit may be coupled with the fluid input port (310) over fluidic test input region (450), and fhiid may be communicated into fluidic channel (452) via the fluid input ports (310, 410) over fluidic test input region (450). This may be done to provide a quality control check, to ensure that layers (400, 500) are properly sealed together. In other words, if back pressure quickly accumulates in the fluid input conduit that is coupled with the fluid input port (310) over fluidic test input region (450), such back pressure may indicate that layers (300, 400) are properly sealed together. If such back pressure does not sufficiently accumulate, this may indicate that layers (400, 500) are not properly sealed together. It should be understood that similar quality control testing may be performed with pneumatic pressurization through a pneumatic port (320) into a pneumatic channel (322), to ensure that layers (300, 400) are properly sealed together.

[6076] A fluidic channel (464) extends between each flush input regions (460) and a corresponding flush output region (462). In some scenarios, as will be described in greater detail below with reference to FIGS. 20A-20B and 21A-21B, a fluid input conduit may be coupled with the fluid input port (310) over flush input regions (460), and a fluid output conduit may be coupled the fluid output port (312) over flush output region (462). Fluid may be communicated into flush input region (460) via fluid input ports (310, 410) over flush input region (460), may flow along fluidic channel (464), then may exit flush output region (462) via fluid output ports (312, 412) over flush output region (462). This may be done to provide flushing of the fluid conduits that are coupled with these ports (310, 412), such as when a cartridge (200) has been replaced, to remove any contaminants that might otherwise be found in those fluid conduits. The combination of a flush input region (460), a fluidic channel (464) a flush output region (462), fluid input port (310), and fluid outlet port (312) may be collectively understood to form a flush assembly. In some examples, the fluid communicated through the flush assembly includes a bio-compatible fluid such as an aqueous based buffer fluid or liquid culture medium; or a bio- compatible oil based fluid. In other examples, other kinds of flush fluid may be used.

[6077] A fluidic channel (604) extends from sample fluid receiving region (600) to a junction (640).

A fluid input conduit may be coupled with the fluid input port (310) over sample fluid receiving region (600), and a fluid containing sample cells may be communicated to junction (640) via the fluid input ports (310, 410) over sample fluid receiving region (600) and via fluidic channel (604). A pair of fluidic channels (626) extend from flow control fluid receiving region (620) to junction (640), such that fluidic channels (604, 626) converge at junction (640). A fluid input conduit may be coupled with the fluid input port (310) over flow control fluid receiving region (620), and a flow control fluid may be communicated to junction (640) via the fluid input ports (310, 410) over flow control fluid receiving region (620) and via fluidic channels (626). The fluid from fluidic channels (604, 626) exits junction (640) along a sampling channel (650).

[6078] The fluid communicated along sampling channel (650) may contain sample cells as noted above. These cells may be imaged by the instrument of system (100) as also noted above.

Such imaging may be performed as the cells traverse sampling channel (650). For instance,

FIG. 6 shows an example of an imaging region (900) that may be positioned along sampling channel (650). In some examples, light source (130) and optical assembly (132) are positioned over cartridge (200) to illuminate imaging region (900); while objective lens assembly (140) and camera (142) are positioned under cartridge (200) to capture images of cells within imaging region (900). Other examples may provide imaging region (900) at any other suitable location or locations along sampling channel (650). Some examples may also provide a plurality of imaging regions (900) at different respective positions along sampling channel (650).

[0079] B. Example of Sorting Features of Cartridge

[0080] As noted above, system (100) may provide sorting of cells in cartridge (120) based on analysis performed by image processing module (144) on images of cells captured by camera (142).

Cartridge (200) of the present example provides a primary sorting region (202) and a secondary sorting region (204) to achieve such sorting.

[6081] FIG. 13 shows primary sorting region (202) of cartridge (200) in greater detail. Primary sorting region (202) is located where sampling channel (650) terminates in a junction (660), which allows fluid to flow from sampling channel (650) to either a first outlet channel (662) or a second outlet channel (664). As shown, a first valve (692) is positioned along first outlet channel (662) while a second valve (690) is positioned along second outlet channel (664).

[0082] First valve (692) is defined by a portion of a pneumatic channel (322b) that is positioned directly over first outlet channel (662), in a region just downstream of junction (660).

Pneumatic port (320b) is pneumatically coupled with pneumatic channel (322b) such that pneumatic channel (322b) may be pressurized via pneumatic port (320b). Second valve (690) is defined by a portion of a pneumatic channel (322a) that is positioned directly over second outlet channel (664). Pneumatic port (320a) is pneumatically coupled with pneumatic channel (322a) such that pneumatic channel (322a) may be pressurized via pneumatic port (3202). Each outlet channel (662, 664) may thus be selectively opened and closed through pneumatic valving via respective valves (692, 690) as described below.

[0083] FIGS. 14A-14C show an example of a sequence of operation of valves (690, 692) during operation of system (100) with cartridge (200). In the state of operation shown in FIG. 14A, neither pneumatic channel (322a, 322b) is pressurized, such that each valve (690, 692) is in an open state. Fluid may thus freely flow through first outlet channel (662) and second outlet channel (664).

[0084] In the state of operation shown in FIG. 14B, pneumatic channel (3222) is pressurized by pressurized air (or any other suitable fluid) that is communicated to pneumatic channel (3222) via pneumatic port (320a). This pressurization of pneumatic channel (322a) causes layer (400) to deform and thereby deflect downwardly in the region underneath pneumatic channel (322a), such that the bottom surface (402) of layer (400) bears against the top surface (502) of layer (500) in the region underneath pneumatic channel (3222). This effectively transitions second valve (690) to a closed state, such that the deformed region of layer (400) under pneumatic channel (322a) prevents fluid from passing through second outlet channel (664). First valve (692) is in the open state in the state of operation shown in FIG. 14B, such that fluid may freely flow through first outlet channel (662).

[0085] In the state of operation shown in FIG. 14C, pneumatic channel (322b) is pressurized by pressurized air {or any other suitable fluid) that is communicated to pneumatic channel (322b) via pneumatic port {320b). This pressurization of pneumatic channel (322b) causes layer (400) to deform and thereby deflect downwardly in the region underneath pneumatic channel (322b), such that the bottom surface (402) of layer (400) bears against the top surface (502) of layer (500) in the region underneath pneumatic channel (322b). This effectively transitions first valve (692) to a closed state, such that the deformed region of layer (400) under pneumatic channel (322b) prevents fluid from passing through first outlet channel (662). Second valve (690) is in the open state in the state of operation shown in FIG. 14C, such that fluid may freely flow through second outlet channel (664).

[0086] When the pneumatic pressure is relieved in the pneumatic channel (322a, 322b) over a given outlet channel (664, 662), the underlying region of layer (400) may return to a non-deformed (e.g., flat) state, thereby effectively opening the fluid pathway through that outlet channel (664, 662). In some examples, this return to the non-deformed state may be provided by resilient properties of layer (400). Each valve (690, 692) may thus be transitioned from an open state to a closed state by pressurizing the corresponding pneumatic channel (322a, 322b); and returned from the closed state to the open state by relieving pressure in the corresponding pneumatic channel (322a, 322b).

[0087] If analysis performed by image processing module (144) on images of cells captured by camera (142) reveals that a cell should be directed to first outlet channel (662) for further sorting via secondary sorting region (204), second valve {690) may be automatically pressurized and thereby transitioned to a closed state while first valve (692) remains in an open state as shown in FIG. 14B. System (100) may then provide further sorting via secondary sorting region (204) as will be described in greater detail below.

[0088] If analysis performed by image processing module (144) on images of cells captured by camera (142) reveals that a cell should be directed to second outlet channel (664) for extraction from cartridge (200) via sample output region (680)), first valve (692) may be automatically pressurized and thereby transitioned to a closed state while second valve (690) remains in an open state as shown in FIG. 14C. The fluid from sampling channel (650) may then flow through outlet channel (664) via junction (660). The fluid path from outlet channel (664) reaches sample output region (680). The fluid exits sample output region (680) via the fluid output ports (412, 312) that are positioned over sample output region (680). A fluid output conduit may be coupled with the fluid output port (412) over sample output region (680), such that the fluid may exit cartridge (200) via this fluid output conduit. The fluid output conduit may be further coupled with a reservoir that is either integrated into the instrument of system (100) or is external to the instrument,

[0089] FIGS. 14-15 show the secondary sorting region (204) of cartridge (200) in greater detail. As shown, secondary sorting region (204) receives fluid via outlet channel (662) from primary sorting region (202). Within secondary sorting region (204), outlet channel (662) branches off into two additional fluidic channels (700, 702) at a junction (704). Fluidic channel (700) branches off into three additional fluidic channels (710, 712, 714) at another junction (706).

Similarly, fluidic channel (708) branches off into three additional fluidic channels (716, 718, 720) at another junction (708). Pneumatic channels (322¢, 322d, 322e, 3221, 322g) pass over fluidic channels (710, 712, 714, 716, 718, 720) within secondary sorting region (204). In this example, pneumatic channels (322c, 322d, 322e, 322f, 322g) all extend along the x-dimension, and fluidic channels (710, 712, 714, 716, 718, 720) all extend along the y-dimension, such that pneumatic channels (322c, 322d, 322e, 322f, 322g) are perpendicular to pneumatic channels (3220, 322d, 322e, 322f, 3229). However, pneumatic channels (322c, 322d, 322e, 322f, 322g) are positioned above fluidic channels (710, 712, 714, 716, 718, 720) along the z-dimension.

[0090] Pneumatic channel (3220) provides a valve (730) over fluidic channel (7 10), a valve (732) over fluidic channel (712), and a valve (734) over fluidic channel (714). Thus, when pneumatic channel (322c) is pneumatically pressurized via a pneumatic port (320c) at the other end of pneumatic channel (322c) (see FIG. 17), the pneumatic pressure causes the region of layer (400) over each fluidic channel (710, 712, 714) to deform downwardly, thereby transitioning each valve (730, 732, 724) to a closed state as described above with respect to valves (690, 692). The closed valves (730, 732, 724) will prevent fluid from flowing through fluidic channels (710, 712, 714) past valves (730, 732, 724). In this example, valves (730, 732, 724) transition between the open and closed state simultaneously. As shown in FIG. 15, pneumatic channel (322¢) is narrower along the y-dimension in the regions where pneumatic channel (322c) passes over fluidic channels (716, 718, 720). This narrowed width of pneumatic channel (322c) prevents deformation of the region of layer (400) over each fluidic channel (716, 718, 720) when pneumatic channel (3220) is pneumatically pressurized. Thus, pneumatic channel {322¢) does not form valves over fluidic channels (716, 718, 720).

[0091] Pneumatic channel (322d) provides a valve (736) over fluidic channel (716), a valve (738) over fluidic channel (718), and a valve (740) over fluidic channel (720). Thus, when pneumatic channel (322d) is pneumatically pressurized via a pneumatic port (320d) at the other end of pneumatic channel (322d) (see FIG. 17), the pneumatic pressure causes the region of layer (400) over each fluidic channel (716, 718, 720) to deform downwardly, thereby transitioning each valve (736, 738, 740) to a closed state as described above with respect to valves (690, 692). The closed valves (736, 738, 740) will prevent fluid from flowing through fluidic channels (716, 718, 720) past valves (736, 738, 740). In this example, valves (736, 738, 740) transition between the open and closed state simultaneously. As shown in FIG. 15, pneumatic channel (322d) is narrower along the y-dimension in the regions where pneumatic channel (322d) passes over fluidic channels (710, 712, 714). This narrowed width of pneumatic channel (322d) prevents deformation of the region of layer (400) over each fluidic channel (710, 712, 714) when pneumatic channel (322d) is pneumatically pressurized. Thus, pneumatic channel (322d) does not form valves over fluidic channels (710, 712, 714).

[0092] Pneumatic channel (322e) provides a valve (752) over fluidic channel (712), a valve (754) over fluidic channel (714), a valve (746) over fluidic channel (716), and a valve (748) over fluidic channel (718). Thus, when pneumatic channel (322e) is pneumatically pressurized via pneumatic port (320e) at the other end of pneumatic channel (322e) (see FIG. 17), the pneumatic pressure causes the region of layer (400) over each fluidic channel (712, 716, 716, 718) to deform downwardly, thereby transitioning each valve (752, 754, 746, 748) to a closed state as described above with respect to valves (690, 692). The closed valves (752, 754, 746,

748) will prevent fluid from flowing through fluidic channels (712, 716, 716, 718) past valves (752, 754, 746, 748). In this example, valves (752, 754, 746, 748) transition between the open and closed state simultaneously. As shown in FIG. 15, pneumatic channel (322e) is narrower along the y-dimension in the regions where pneumatic channel (322e) passes over fluidic channels (710, 720). This narrowed width of pneumatic channel (322e) prevents deformation of the region of layer (400) over each fluidic channel (710, 720) when pneumatic channel (3226) is pneumatically pressurized. Thus, pneumatic channel (322e) does not form valves over fluidic channels (710, 720).

[0093] Pneumatic channel (322f) provides a valve (770) over fluidic channel (710), a valve (774) over fluidic channel (714), a valve (786) over fluidic channel (716), and a valve (790) over fluidic channel (720). Thus, when pneumatic channel (322f) is pneumatically pressurized via a pneumatic port (320f) at the other end of pneumatic channel (322f) (see FIG. 17), the pneumatic pressure causes the region of layer (400) over each fluidic channel (710, 714, 716, 720) to deform downwardly, thereby transitioning each valve (770, 774, 776, 790) to a closed state as described above with respect to valves (690, 692). The closed valves (770, 774, 776, 790) will prevent fluid from flowing through fluidic channels (710, 714, 716, 720) past valves (770, 774, 776, 790). In this example, valves (770, 774, 776, 790) transition between the open and closed state simultaneously. As shown in FIG. 15, pneumatic channel (322f) is narrower along the y- dimension in the regions where pneumatic channel (322f) passes over fluidic channels (712, 718). This narrowed width of pneumatic channel (322f) prevents deformation of the region of layer (400) over each fluidic channel (712, 718) when pneumatic channel (322f) is pneumatically pressurized. Thus, pneumatic channel (322f) does not form valves over fluidic channels (712, 718).

[0094] Pneumatic channel (322g) provides a valve (771) over fluidic channel (710), a valve (773) over fluidic channel (712), a valve (798) over fluidic channel (718), and a valve (791) over fluidic channel (720). Thus, when pneumatic channel (322g) is pneumatically pressurized via a pneumatic port (320e) at the other end of pneumatic channel (322g) (see FIG. 17), the pneumatic pressure causes the region of layer (400) over each fluidic channel (710, 712, 718, 720) to deform downwardly, thereby transitioning each valve (771, 773, 798, 791) to a closed state as described above with respect to valves (690, 692). The closed valves (771, 773, 798, 791) will prevent fluid from flowing through fluidic channels (710, 712, 718, 720) past valves (771, 773, 798, 791). In this example, valves (771, 773, 798, 791) transition between the open and closed state simultaneously. As shown in FIG. 15, pneumatic channel (322g) is narrower along the y-dimension in the regions where pneumatic channel (322g) passes over fluidic channels (714, 716). This narrowed width of pneumatic channel (322g) prevents deformation of the region of layer (400) over each fluidic channel (714, 716) when pneumatic channel (322g) is pneumatically pressurized. Thus, pneumatic channel (322g) does not form valves over fluidic channels (714, 716).

[0095] As described above, when a pneumatic channel (322c, 322d, 322e, 322f, 322g) is in a pneumatically pressurized state, the valves (730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 7798, 791) of that pneumatic channel (322¢, 322d, 322e, 322f, 3229) will be in a closed state. When a pneumatic channel (322c, 322d, 322e, 322f, 322g) is in a non-pressurized state, the valves (730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791) of that pneumatic channel (3220, 322d, 322e, 322f, 3229) will be in an open state. Fluid will be allowed to flow through, or prevented from flowing through, each fluidic channel (710, 712, 714, 716, 718, 720), and thereby reach the corresponding well (750, 752, 754, 756, 758, 760), based on whether all the valves (730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791) along the fluidic channel (710, 712, 714, 716, 718, 720) are in an open state. If one or more valves (730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791) along a fluidic channel (710, 712, 714, 716, 718, 720) is/are in a closed state, then fluid may not flow through fluidic channel (710, 712, 714, 716, 718, 720) and reach the corresponding well (750, 752, 754, 756, 758, 760).

[0096] The following table provides an example of the different pneumatic states that may be provided in pneumatic channels (322c, 322d, 322e, 322f, 3229) to permit or prevent fluid (and a cell contained therein) to flow to each well (750, 752, 754, 756, 758, 760):

TABLE

Fluid/Cell State of State of State of State of State of

Destination channel channel channel channel channel (3220) (322d) (322e) (322f) (3229) well (750) non- pressurized pressurized non- non-

Eh ree] [ee well (754) non- pressurized non- non- pressurized

ET well (756) pressurized non- non- non- pressurized naa well (758) pressurized non- non- pressurized non-

B well (760) pressurized non- pressurized non- non-

[0097] In some scenarios, it may be beneficial to provide additional assistance to the flow of fhad from outlet channel (662) through whichever fluidic channel (710, 712, 714, 716, 718, 720) is selected for routing via secondary sorting region (204) to reach a corresponding selected well (750, 752, 754, 756, 758, 760). To provide such assistance, additional fluid may be communicated through a fluid input conduit that is coupled with the fluid input port (310) over flow drive fluid receiving region (670) of cartridge (200). This additional fluid may reach outlet channel (662) via ports (310, 410) over flow drive fluid receiving region (670) and further via fluidic channel (672). At the time this additional fluid is communicated to outlet channel (662), the pneumatic valve between outlet channel (662) and junction (660) may be in a closed state. As shown in FIG. 17, a pneumatic valve (795) is positioned along fluidic channel (672) downstream of flow drive fluid receiving region (670). Pneumatic valve (795) is closed through pneumatic pressurization of pneumatic channel (322h) via pneumatic port (320h).

[0098] The additional fluid from flow drive fluid receiving region (670) and fluidic channel (672) may provide a “boost” to fluid in outlet channel (662), thereby further driving the fluid from outlet channel (662) through the selected fluidic channel (710, 712, 714, 716, 718, 720); and ultimately to the selected well (750, 752, 754, 756, 758, 760). Whichever valves (730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791) are open to allow the sample cell carrying fluid to flow from outlet channel (662) to the selected well (750, 752, 754, 756, 758, 760) may remain open as needed to allow the boosting fluid from flow drive fluid receiving region (670) to adequately flow through the fluidic channel (710, 712, 714, 716, 718, 720) corresponding to the selected well (750, 752, 754, 756, 758, 760).

[0099] System (100) may execute a control algorithm to automatically select which pneumatic valves (690, 692, 730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791) to activate. This control algorithm may be executed in response to data from image processing module (144). In other words, image processing module (144) may classify or otherwise analyze images of cells captured by camera (142) as the cell-containing fluid passes through imaging region (900), and the pneumatic valves 690, 692, 730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774,786, 790, 771, 773, 798, 791) described above may provide sorting of cells in the fluid based on such image processing. For instance, cells of a first type (as identified by image processing module (144)) may be routed via pneumatic valving at primary and secondary sorting regions (202, 204) to a first well (750), cells of a second type (as identified by image processing module (144)) may be routed via pneumatic valving at primary and secondary sorting regions (202, 204) to a second well (752), and cells of a third type (as identified by image processing module (144)) may be routed via pneumatic valving at primary sorting region (202) to sample output region (680).

[00100] C. Example of Flow Control Features of Cartridge

[60101] As noted above, a fluid containing sample cells may be communicated to cartridge (200) via a fluid input conduit that is positioned over sample fluid receiving region (600); while a flow control fluid may be communicated to cartridge (200) via a fluid input conduit that is positioned over flow control fluid receiving region (620). FIG. 7 shows these regions (600, 620) in greater detail. In some examples, the flow control fluid includes a buffer fluid. In some examples the flow control fluid further includes beads; while in other examples the flow control fluid does not include beads. The flow control fluid may include a bio-compatible fluid. In some examples, the flow control fluid has a viscosity that differs from the viscosity of the fluid containing sample cells. In other examples, other kinds of fluid may be used for flow control fluid.

[00102] As shown, sample fluid receiving region (600) comprises a teardrop shaped recess in the bottom surface (404) of layer (400), with an array of filtering elements (602) positioned within the teardrop shape. As best seen in FIG. §, each filtering element (602) in this example is in the form of a cylindraceous structure that extends down to top surface (502) of layer (500).

Filtering elements (602) are spaced apart from each other with enough distance to allow cells to be carried by the cell-containing fluid through the recess of fluid receiving region (600) to fluidic channel (604). However, filtering elements (602) are close enough to each other to filter out any debris that may be present in the cell-containing fluid, such that filtering elements (602)

may prevent unwanted clogging of fluidic channel (604).

[60103] Flow control fluid receiving region (620) comprises a circle shaped recess in the bottom surface (404) of layer (400), with an array of filtering elements (622) positioned within the circle shape.

Filtering elements (622) of flow control fluid receiving region (620) are structurally configured and operable just like filtering elements (602) described above. Filtering elements (622) may thus prevent unwanted clogging of a fluidic channel (624), which provides a pathway from flow control fluid receiving region (620) to fluidic channels (626). As shown in FIG. 7, fluidic channels (626) include a first fluidic channel (626a) and a second fluidic channel (626b), which have mirror symmetry with each other. Each fluidic channel (626) has a serpentine configuration with a herringbone region (630) and an outlet region (638) leading to junction (640).

[00104] While sample fluid receiving region (600) and flow control fluid receiving region (620) are described above as including filtering elements (602, 622), other portions of cartridge (200) may also include filtering elements like filtering elements (602, 622). For instance, fluidic test i5 input region (450), flush input regions (460), flush output regions (462), flow drive fluid receiving region (670), and/or sample output region (680) may each include filtering elements like filtering elements (602, 622).

[00105] In some examples, it may be desirable to carefully control the fluid flow dynamics within sampling channel (650) during operation of system (100) with cartridge (200). Controlling the fluid flow dynamics within sampling channel (650) may ensure that the cells in in the fluid communicated along sampling channel (650) have a desired spacing relative to each other and/or relative to the three-dimensional confines of sampling channel (650). Providing such control over the spacing of the cells relative to each other and/or relative to the three- dimensional confines of sampling channel (650) may optimize the imaging of the cells within imaging region (900), which may in turn optimize the processing of the cell images and the results obtained through such image processing. The following description provides examples of how the fluid flow dynamics within sampling channel (650) may be controlled.

[00106] D. Examples of Mixing Structures in Side Channels Upstream of Junction

[00107] In some examples, it may be desirable to perform a calibration of camera (142) and objective lens assembly (140) before capturing images of cells in imaging region (900). In some of those examples, it may be desirable to communicate beads (or other particles) having a known structural configuration along sampling channel (650), such that camera (142) may capture images of the beads as they pass through imaging region (900) during a calibration routine. In some examples, such beads (or other particles) may also be communicated along sampling channel (650) while sample cells (or other sample particles) are also communicated along sampling channel (650). The presence of the beads in sampling channel (650) may allow image processing module (144) to detect changes in the structural configuration of sampling channel (650) during operation of system (100), such as expansion of sampling channel (650) caused by thermal expansion of cartridge (200) during operation of system (100). In some such examples, such changes in the structural configuration of sampling channel (650) during operation of system (100) may be detected due to a tendency of beads to flow along separate x-y planes that are separated from each other along the z-dimension by a distance that varies with the structural configuration of sampling channel (650).

[00108] In some cases where beads are used, it may be beneficial to communicate those beads through fluidic channels (626). Those beads may tend to arrange themselves within the fluid flowing along fluidic channels (626) in a predictable arrangement. Even though it may be desirable to have a predictable arrangement of particles (e.g., cells) as those particles travel along sampling channel (650) in some examples, it may be desirable for particles to have an unpredictable arrangement before entering junction (640) in some examples. An unpredictable arrangement of particles upstream of junction (640) may in fact lead to a more predictable arrangement of particles downstream of junction (640). It may therefore be desirable in some examples to provide a feature in fluidic channels (626) that will add chaos to the arrangement of beads in fluid that flows through fluidic channels (626).

[00109] As noted above, each fluidic channel (626) includes a herringbone region (630). FIGS. 9-11 show herringbone region (630) in greater detail. Herringbone region (630) serves as a mixing feature and thus provides chaos to the arrangement of beads in fluid that flows through herringbone region (630). Each herringbone region (630) includes a plurality of chevron shaped recesses (634) above the flow channel (632) through herringbone region (630). Each recess (634) includes a ceiling (634) and set of sidewalls (636). In some examples, as shown in FIG. 11, sidewalls (636) define a length (1.2) of approximately 60 um. Sidewalls (636) may also form angles (©) with each other at approximately 45 degrees. Ceiling (634) may be positioned at a height (H3) of approximately 33 um relative to the bottom of each sidewall (636). The specific dimensions provided above are not intended to be limiting. Other dimensions may be used.

[00110] FIG. 12A shows an example of an arrangement of beads (652) in fluidic channel (626) upstream of herringbone region (630); while FIG. 12B shows an example of an arrangement of beads (652) in fluidic channel (660) in outlet region (638) of fluidic channel (626) downstream of herringbone region (630). As shown, the arrangement of beads (652) upstream of herringbone region (630) is substantially orderly; whereas the arrangement of beads (652) downstream of herringbone region (630) is substantially stochastic or randomized. This stochastic or randomized arrangement of beads (652) in outlet region (638) may provide a greater likelihood that beads (652) will be subject to the flow focusing phenomenon described herein than might otherwise be achieved in the absence of herringbone region (630). In other words, without herringbone region (630), beads (652) may be less likely to mimic the arrangement in imaging region (900) that would be achieved by cells later passing through imaging region (900).

Herringbone region (630) may thus facilitate replication of cell arrangements in imaging region (900) by bead (660) arrangements in imaging region (900) during calibration of system (100).

[00111] While FIG. 7 shows each fluidic channel {626) having only one herringbone region (630), each fluidic channel (626) may have two or more herringbone regions (630) in other examples.

Some other examples may omit herringbone regions (630).

[60112] E. Examples of Flow Resistance Relationships in Channels of Junction

[00113] In some examples, it may be desirable to maximize the stability of the flow rate of fluid along the entire length of sampling channel (650). For instance, in cases where pneumatic valves are actuated downstream of junction (660) to sort cells in response to image analysis of the cells as they pass through imaging region (900), a stable flow rate of fluid will ensure that the cells are sorted as intended. In other words, a stable flow rate through sampling channel (650) may ensure appropriate synchronization between valve timing and image capture/analysis. The inclusion of flow focusing features such as those described below may tend to complicate the achievement of a stable flow rate through sampling channel (650).

[00114] In some examples, it may also be desirable to allow the flow rate of fluid being input through fluid input port (410) of sample fluid receiving region (600), and to allow the flow rate of fluid being input through fluid input port (410) of flow control fluid receiving region (620) to be adjusted, while still providing a stable flow rate through sampling channel (650). Allowing greater flexibility in input fluid flow rates may increase the versatility of cartridge (200), such as by allowing end users to meet different fluid flow requirements based on the process protocols at hand.

[00115] The above-described results may be achieved by providing tuned flow resistance through fluidic channels (604, 626, 650) as described below. In particular, the stability of flow along sampling channel (650) is provided or promoted, in part, through precise control of the flow rate of fluid entering junction (640). In turn, the control of the flow rate of fluid entering junction (640) is provided, in part, by providing tuned flow resistance through fluidic channel (604) and each outlet region (638) of each fluidic channel (626). By tuning the flow resistance through fluidic channel (604) and each outlet region (638) of each fluidic channel (626), cartridge (200) may accommodate different flow rates of fluid received via sample fluid receiving region (600) and flow control fluid receiving region (620), while still maintaining a certain flow rate ratio through fluidic channel (604) and each outlet region (638). This may allow cartridge (200) to accommodate different end user needs, which may include different flow rates and driving pressures through sample fluid receiving region (600) and flow control fluid receiving region (620).

[00116] The pressure of sample-carrying fluid communicated through fluidic channel (604) may be expressed as “P;,” the flow resistance of fluidic channel (604) may be expressed as “R;,” and the flow rate through fluidic channel (604) may be expressed as “7,” The pressure of flow control fluid communicated through each fluidic channel (626) may be expressed as “P»,” the flow resistance of each fluidic channel (626) may be expressed as “R.,” and the flow rate through each fluidic channel (626) may be expressed as “I>.” The pressure of the combined fluid communicated through sampling channel (650) may be expressed as “Po,” the flow resistance of sampling channel (650) may be expressed as “Ro,” and the flow rate through sampling channel (650) may be expressed as “ly.” With these parameters identified, a ratio of the flow rate /; of fluidic channel (604) to the flow rate I; of each fluidic channel (626) may be expressed in the following equation (1): a I _ Ry(Py=Po)+Ry(P1=P;)

I, Ry(Pa~Po)+Ro(Py~Py)

[00117] The flow rate I, of sampling channel (650) may be expressed in the following equation (II): a b= hn,

[60118] One or more pumps (110) and/or other components may be operated to drive the fluids through fluidic channels (604, 626, 650) may achieve a desired main flow rate 1, in sampling channel (650), and ratio of flow rates 1/1; of fluidic channels (604, 626), based on the two equations (1,

II) above. As can be seen above, these equations (I, II) factor in the flow resistance values R;,

R>, Ro of fluidic channels (604, 626, 650). The outcomes of these equations (1, 11) may thus be influenced by how fluidic channels (604, 626, 650) are structurally configured to tune the flow resistance value R;, Ra, Ro of each fluidic channel (604, 626, 650), since the flow resistance value R;, Az, Ro of each fluidic channel (604, 626, 650) may be fixed by the structural configuration of each fluidic channel (604, 626, 650).

[00119] In the present example, channels (604, 626, 650) are structurally configured such that the flow resistance R; of fluidic channel (604) is substantially lower than the flow resistances A», Roof channels (626, 650); and such that the flow resistance R; of each fluidic channel (626) is substantially equal to (or on the same order as) the flow resistance Ry of sampling channel (650). This relationship may thus be expressed in in the following equation (II):

[00120] (ID) Ri K Ry, ~Ry

[00121] By way of example only, the value of flow resistance R; of fluidic channel (604) may range from approximately 1/5 of the value of the flow resistance Ry of sampling channel (650) to approximately ¥2 of the value of the flow resistance Ry of sampling channel (650). Similarly, the value of flow resistance A; of fluidic channel (604) may range from approximately 1/5 of the value of the flow resistance R; of each fluidic channel (626) to approximately 14 of the value of the flow resistance R2 of each fluidic channel (626).

[00122] Channels (604, 626, 650) may be structurally configured such that the flow resistance R; of fluidic channel (604) is substantially lower than the flow resistances R>, Ro of channels (626, 650) by fluidic channel (604) having a substantially shorter length than the length of channels (626, 650). Similarly, channels (604, 626, 650) may be structurally configured such that the flow resistance R;of each fluidic channel (626) is substantially equal to (or on the same order as) the flow resistance Ro of sampling channel (650) by each fluidic channel (626) having a length that is substantially equal to (or on the same order as) the length of sampling channel (650). To achieve substantially equal length of each fluidic channel (626) and sampling channel (650) without creating unnecessary overall length in cartridge (200), each fluidic channel (626) has a serpentine configuration. In some examples, the different flow resistances

Ri, R>, Ro may be achieved by using different cross-sectional areas {(e.g., different heights and/or widths) among channels (604, 626, 650).

[00123] With the relationships between R;, R>, Ro provided as expressed in equation (Il), the ratio of the flow rate 7; of each fluidic channel (626) to the flow rate £ of fluidic channel (604) may be simplified as the following equation (IV):

(IV) fo ~ 2

[00124] Similarly, with the relationships between R;, R2, Ry as expressed in equation (HI), the flow rate

Io of sampling channel (650) may simplified as the following equation (IV): 3 av) oe

[00125] The substantially lower flow resistance R; of fluidic channel (604) may be due, at least in part, to fluidic channel (604) having a substantially shorter length than the lengths of channels (626, 650). In some examples, the flow resistance R: of each fluidic channel (626) ranges from approximately 10 times the flow resistance R; of fluidic channel (604) to approximately 50 times the flow resistance R; of fluidic channel (604); or more particularly, may be approximately 20 times the flow resistance R; of fluidic channel (604). In some examples, the flow resistance of sampling channel (650) ranges from being approximately equal to the flow resistance R; of each fluidic channel (626) to being approximately 5 times the flow resistance

R: of each fluidic channel (626); or more particularly, may be approximately 3 times the flow resistance R; of each fluidic channel (626).

[00126] Minimizing the flow resistance R; of fluidic channel (604) may minimize the fluid pressure P; applied through sample fluid receiving region (600) and fluidic channel (604), which may in turn minimize the risk of damage to cells contained in the fluid that is communicated through sample fluid receiving region (600) and fluidic channel (604). Having a higher pressure P: of fluid in fluidic channels (626) (e.g., due to the substantially higher flow resistance of fhiidic channels (626)) may not be problematic since the fluid communicated through flow control fluid receiving region (620) and fluidic channels (626) does not contain cells that might otherwise be damaged. Thus, in some examples of operation, the pressure P; of fluid in fluidic channels (626) is set to be higher than the pressure P; of fluid in fluidic channel (604). In addition, to provide positive inflow from fluidic channel (604) into junction (640), the pressure

P; of fluid in fluidic channels (626) and the pressure P; of fluid in fluidic channel (604) may be controlled to adhere to the following equation (V):

[00127] F. Example of Ventilation Features in Cartridge

[00128] The air or other gas that is used to provide pneumatic pressurization in pneumatic channels (322) will hereinafter be referred to as “pneumatic pressure gas.” In some examples, after one or more pneumatic channels (322) have been pressurized during use of cartridge (200), it may be possible that some of the pneumatic pressure gas may tend to enter the space between layers (300, 400), between bottom surface (320) and top surface (402). This may tend to create separation between layers (300, 400). In some examples, the pneumatic pressure gas may tend to build up in one or more of pneumatic channels (322) and/or migrate from one pneumatic channel (322) to another pneumatic channel (322) after cartridge (200) is used for some time.

Even if the pneumatic pressure gas does not migrate through the space between layers (300, 400) or otherwise cause separation between layers (300, 400), in some examples the material forming layer (400) may allow some pneumatic pressure gas to pass through layer (400) via diffusion. Regardless of whether the pneumatic pressure gas diffuses through layer (400) and/or between layers (300, 400), the pneumatic pressure gas may eventually reach one or more fluidic channels such as sampling channel (650). One or more of the above-described effects may occur in some cases where one or more pneumatic valves of cartridge (200) is/are maintained in a closed state for a long period of time and/or in other scenarios.

[00129] To prevent or mitigate the risk of pneumatic pressure gas passing through layer (400) and/or between layers (300, 400), cartridge (200) of the present example include venting features that allow the pneumatic pressure gas to escape to atmosphere. These venting features do not adversely affect the ability of the pneumatic pressure gas to achieve successful operation of the valves of cartridge (200) as described above. Venting features of the present example are shown in FIGS. 17-18, and include a set of vent passageways (800, 810) and vent channels (802, 804). Vent passageways (800, §10) extend through the full thickness of layer (300) along the z-dimension, such that each vent passageway (800, 810) is open at the top surface (302) and at the bottom surface (304). Each vent passageway (800, 810) further provides a path to the atmosphere to allow venting to the atmosphere.

[00130] Each vent channel (802, 804) is formed as a recess in bottom surface (304). Vent channels (802) extend directly from vent passageway (800) along the x-y plane; and vent channels (804) branch off from vent channels (802) along the x-y plane. Top surface (402) of layer (400) defines a bottom of each vent channel (802, 804) in this example. Any pneumatic pressure gas that reaches vent channels (802, 804) (e.g., through layer (400) and/or through a space between layers (300, 400)) may be relieved to the atmosphere along the space defined collectively by top surface (502) of layer (500) and each vent channel (802, 804), with such space eventually reaching vent passageway (800).

[00131] As shown in FIGS. 17-18, each vent channel (802) is positioned to be near one or more pneumatic channels (322). In particular, vent channel (804a) is positioned near pneumatic channel (322h); vent channel {804b) is positioned between pneumatic channels (322h, 322¢); vent channel (804c) is positioned between pneumatic channels (322c, 322d); vent channel (804d) is positioned between pneumatic channels (322d, 322¢); vent channel (804e) is positioned between pneumatic channels (322d, 322e); vent channel (804f) is positioned between pneumatic channels (322e, 322f); and vent channel (804g) is positioned near pneumatic channels (804g). Vent channel (804a) is this positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from pneumatic channel (322h); vent channel (804b) is positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (322h, 322¢); vent channel (804c) is positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (322¢, 322d); vent channel (804d) is positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (322d, 322¢); vent channel (804e) is positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (322d, 322e); vent channel {804f) is positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (322e, 322f); and vent channel (804g) is this positioned to provide venting for pneumatic pressure gas that may tend to otherwise build up from pneumatic channel (804g).

[00132] As described above, vent passageway (800) is positioned to relieve to atmosphere any pneumatic pressure gas that reaches any of vent channels (802, 804). Vent passageway (810) does not have any associated vent channels in this example, though vent passageway (810) may have one or more associated vent channels in some other examples. Even without any associated vent channels, vent passageway (810) may still effectively relieve to atmosphere any pneumatic pressure gas that reaches vent passageway via layer (400) or via the space between layers (300, 400).

[00133] FIG. 19 shows an example of another arrangement that may be used to provide ventilation in a cartridge (102) of system (100). Specifically, FIG. 19 shows features of another cartridge that includes a sampling channel (1650) that is like sampling channel (650), and a plurality of pneumatic channels (1322) that are like pneumatic channels (322). Specifically, pneumatic channel {1322b) is like pneumatic channel (322b), pneumatic channel (1322¢) is like pneumatic channel (322c¢), pneumatic channel (1322d) is like pneumatic channel (322d), pneumatic channel (1322e) is like pneumatic channel (322e), pneumatic channel (13221) is like pneumatic channel (322f), and pneumatic channel (1322g) is like pneumatic channel (322g). The cartridge venting arrangement of FIG. 19 also includes a plurality of vent passageways (1810) that are like vent passageway (810).

[00134] Vent passageway (1810a) is positioned near pneumatic channels (1322b, 1322h), such that vent passageway (1810a) may provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (1322b, 1322h). Vent passageway (1810b) is positioned near pneumatic channels (1322h, 1322c¢), such that vent passageway (1810b) may provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (1322h, 1322c). Vent passageway (1810c) is positioned near pneumatic channels (1322d, 1322e), such that vent passageway (1810c) may provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (1322d, 1322e). Vent passageway (1810d) is positioned near pneumatic channels (1322¢, 1322f), such that vent passageway (1810d) may provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (1322e, 1322f). Vent passageway (18106) 1s positioned near pneumatic channels (1322f, 1322g), such that vent passageway (1810e) may provide venting for pneumatic pressure gas that may tend to otherwise build up from either or both of pneumatic channels (1322f, 1322g). Vent passageway (1810f) is positioned near pneumatic channel (13229), such that vent passageway (1810f) may provide venting for pneumatic pressure gas that may tend to otherwise build up from pneumatic channel (1322g).

[00135] While vent passageways (1810) of FIG. 19 do not have any corresponding vent channels like vent channels (802, 804), some other examples may include one or more vent channels that allow pneumatic pressure gas to flow toward one or more of vent passageways (1810) of FIG. 19.

[00136] G. Example of Conduit Flush Ports in Cartridge

[00137] In some examples, system (100) may be used in different processes where one or more fluids are communicated through one or more cartridges (200) in different sessions of use. Some examples of system (100) may include integral fluid conduits that are reused with the same cartridge (200) and/or with different cartridges during different sessions of use of system (100).

In such scenarios, such fluid conduits may be cleaned between different sessions of use of system (100), to avoid or mitigate any risk of contamination from residual fluids or materials that might otherwise remain in such fluid conduits between sessions of use of system (100). In cartridge (200), flush input regions (460) and flush output regions (462) may be used to flush fluid conduits that are otherwise used to communicate fluids to or from cartridge (200).

[00138] FIG. 20A shows an arrangement where a first conduit (820) is coupled with sample fluid receiving region (600) and with a source (824) of a fluid containing one or more sample cells.

For instance, source (824) may constitute pump (110) and reservoir (112) of system (100) as described above. In the present example, first conduit (820) comprises a flexible tube that is inserted into fluid input port (310a) directly over sample fluid receiving region (600), with the material of layer (300) providing a friction fit to retain and seal first conduit (820) relative to cartridge (200). A second conduit (822) is coupled with flow control fluid receiving region (620) and with a source (826) of flow control fluid. In some examples, the flow control fluid includes a buffer fluid. In some examples the flow control fluid further includes beads; while in other examples the flow control fluid does not include beads. The flow control fluid may include a bio-compatible fluid. In some examples, the flow control fluid has a viscosity that differs from the viscosity of the fluid containing sample cells. In other examples, other kinds of fluid may be used for flow control fluid. In the present example, second conduit (822) comprises a flexible tube that is inserted into fluid input port (310b) directly over flow control fluid receiving region (620), with the material of layer (300) providing a friction fit to retain and seal second conduit (822) relative to cartridge (200).

[00139] System { 100) may be operated with cartridge (200) and conduits (820, 822) in the state shown in FIG. 20A, as described herein, to capture images of sample cells, classify the imaged sample cells, and sort the sample cells. Once this session of use is complete (e.g., after all of the sample cells from source (824) have been processed), an operator may pull first conduit (820) out of fluid input port (310a) and pull second conduit (822) out of fluid input port (310b). The operator may then couple first conduit (820) with flush output region {462a) by inserting first conduit (820) into fluid output port (312a) directly over flush output region (462a). The material of layer (300) may provide a friction fit to retain and seal first conduit (820) relative to cartridge (200) at fluid output port (312a). The other end of first conduit (820) may be decoupled from source (824) and coupled with a waste reservoir (825). The operator may also couple second conduit (822) with flush input region (460a) by inserting second conduit (822) into fluid input port (310c) directly over flush input region (4602). The material of layer (300)

may provide a friction fit to retain and seal second conduit (822) relative to cartridge (200) at fluid input port (310c). The other end of second conduit (822) may be decoupled from source (826) and coupled with a source (827) of flush fluid. An example of the resulting arrangement is shown in FIG. 20B.

[00140] As noted above, a fluidic channel (464a) extends between flush input region {4602) and flush output region (462a). Flush fluid from source (827) may thus be communicated through second conduit (822) to flush input region (460a), through fluidic channel (464a), out flush output region (462a) and along first conduit (820) to waste reservoir (825). In some examples, the flush fluid is pushed through this fluid circuit from the source (827) side of the fluid circuit, such as by a pump. In some other examples, the flush fluid is pulled through this fluid circuit from the waste reservoir (825) side of the fluid circuit. In either of these arrangements, the wash fluid may flow through both conduits (820, 822), with cartridge (200) providing a connection path for the wash fluid to flow between conduits (820, 822). After conduits (820, 822) have been sufficiently flushed, conduits (820, 822) may be returned to the arrangement shown in FIG. 20A.

[00141] While first conduit (820) is coupled with flush output region (462a) and second conduit (822) is coupled with flush input region (460a) during the flushing stage shown in FIG. 20B and described above, in other examples second conduit (822) may be coupled with flush output region (462a) and first conduit (820) may be coupled with flush input region (460a) during the flushing stage.

[00142] FIG. 21A shows an arrangement where a third conduit (830) is coupled with flow drive fluid receiving region (670) and with a source (834) of flow drive fluid. In some examples, the same fluid that is communicated to flow control fluid receiving region (620) for flow control in cartridge (200) is also communicated to flow drive fluid receiving region (670) to boost fluid flow through and past secondary sorting region (204). In some such examples, source (824) and source (834) may be the same source. In some examples, the additional boosting fluid provided via flow drive fluid receiving region (670) and fluidic channel (672) may include any bio-compatible fluid, such as liquid culture medium, cell lysis solution, or an oil based fluid.

In other examples, other kinds of fluid may be used.

[00143] In the present example, third conduit (830) comprises a flexible tube that is inserted into fluid input port (310c) directly over flow drive fluid receiving region (670), with the material of layer (300) providing a friction fit to retain and seal third conduit (830) relative to cartridge

(200). A fourth conduit (832) is coupled with sample output region (680) and with a receptacle (836) (e.g., vial, tube, etc.) that receives fluid communicated from cartridge (200) via sample output region (680). In the present example, fourth conduit (832) comprises a flexible tube that is inserted into fluid output port (312c) directly over sample output region (680), with the material of layer (300) providing a friction fit to retain and seal fourth conduit (832) relative to cartridge (200).

[09144] System (100) may be operated with cartridge (200) and conduits (830, 832) in the state shown in FIG. 21A, as described herein, to capture images of sample cells, classify the imaged sample cells, and sort the sample cells. Once this session of use is complete (e.g., after all of the sample cells from source (824) have been processed), an operator may pull third conduit (830) out of fluid input port (310c) and pull fourth conduit (832) out of fluid output port (312c). The operator may then couple third conduit (830) with flush output region {462b) by inserting third conduit (830) into fluid output port (312b) directly over flush output region (462b). The material of layer (300) may provide a friction fit to retain and seal third conduit (830) relative to cartridge {200) at fluid output port (312b). The other end of third conduit (830) may be decoupled from source (834) and coupled with a waste reservoir (835). An example of the resulting arrangement is shown in FIG. 21B. In some examples, waste reservoir (835) is the same as waste reservoir (825). In examples where the same waste reservoir (825, 835) to receive waste from first and third conduits (820, 830), the fluidic path between first conduit (820) and the shared reservoir (825, 835) may differ from the fluidic path between third conduit (830) and the shared reservoir (825, 835).

[00145] The operator may also couple fourth conduit (832) with flush input region (460b) by inserting fourth conduit (832) into fluid input port (310d) directly over flush input region (460b). The material of layer (300) may provide a friction fit to retain and seal fourth conduit (832) relative to cartridge (200) at fluid input port (310d). The other end of fourth conduit (832) may be decoupled from receptacle (836) and coupled with a source (827) of flush fluid. An example of the resulting arrangement is shown in FIG. 21B. In some examples, source (837) is the same as source (827). In examples where the same source (827, 837) to is used to provide flush fluid to second and fourth conduits (822, 832), the fluidic path between second conduit (822) and the shared source (827, 837) may differ from the fluidic path between fourth conduit (832) and the shared source (827, 837).

[00146] As noted above, a fluidic channel (464b) extends between flush input region (460b) and flush output region (462b). Flush fluid from source (837) may thus be communicated through fourth conduit (832) to flush input region (460b), through fluidic channel {464b), out flush output region (462b) and along third conduit (830) to waste reservoir (835). In some examples, the flush fluid is pushed through this fluid circuit from the source (837) side of the fluid circuit, such as by a pump. In some other examples, the flush fluid is pulled through this fluid circuit from the waste reservoir (835) side of the fluid circuit. In either of these arrangements, the wash fluid may flow through both conduits (830, 832), with cartridge (200) providing a connection path for the wash fluid to flow between conduits (830, 832). After conduits (830, 832) have been sufficiently flushed, conduits (830, 832) may be returned to the arrangement shown in FIG. 21A.

IO [00147] While third conduit (830) is coupled with flush output region (462b) and fourth conduit (832) is coupled with flush input region (460b) during the flushing stage shown in FIG. 21B and described above, in other examples fourth conduit (832) may be coupled with flush output region (462b) and third conduit (830) may be coupled with flush input region (460b) during the flushing stage.

[00148] H. Example of Visual Fiducials and Focusing Features of Cartridge

[00149] As noted above, objective lens assembly (140) and camera (142) capture images of cells conveyed via fluid along sampling channel (650), with such images being captured within imaging region (900). In some examples it may be beneficial to ensure that cartridge (200) is appropriately positioned in relation to objective lens assembly (140) and camera (142), to ensure that objective lens assembly (140) and camera (142) may adequately capture images of cells conveyed via fluid along sampling channel (650). This may include ensuring that cartridge (200) is appropriately seated in a cartridge mount of the instrument of system (100); that the cartridge mount is appropriately positioned within the instrument relative to objective lens assembly (140) and camera (142); and/or that objective lens assembly (140) and camera (142) are otherwise appropriately positioned relative to cartridge (200).

[00150] FIGS. 22-26 show an example optical fiducial region (840) of cartridge (200). Optical fiducial region (840) is positioned near imaging region (900) in this example. Optical fiducial region (840) includes a plurality of fiducial elements (844) arranged in a rectangle with four corners {842a, 842b, 842c, 842d). Fiducial elements (844) include recesses in bottom surface (404) of layer (400), such that fiducial elements (844) have optically detectable edges extending along the z-dimension, thereby defining optically detectable shapes along the x-y plane. These shapes include ellipses.

[00151] The elliptical shape of each fiducial element (844) differs from the elliptical shape of the other fiducial elements (844). For instance, as best seen in FIG. 23, fiducial elements (844) in the upper left-hand corner (842a) of optical fiducial region (840) have a major axis extending along the x-dimension, with the minor axis extending along the y-dimension. While the elliptical shapes of fiducial elements (844) in the upper left-hand corner (8422) of optical fiducial region (840) share this general characteristic, the lengths of the major and minor axes still vary among fiducial elements (844) in the upper left-hand corner (8422).

[00152] As best seen in FIG. 24, fiducial elements (844) approach a circular shape at the upper right- hand corner (842b) of optical fiducial region (840). The length of the axis along the x- dimension of the elliptical shape for each fiducial element (844) gradually reduces per each fiducial element (844) as the array of fiducial elements (844) spans along the x-dimension, such that fiducial elements (844) have a longest axis along the x-dimension in the left-hand side of optical fiducial region (840) and the shortest axis along the x-dimension in the right-hand side of optical fiducial region (840). i5 [00153] As best seen in FIG. 25, which shows the lower left-hand corner (842c) of optical fiducial region (840), the length of the axis along the x-dimension of the elliptical shape for each fiducial element (844) remains the same per each fiducial element (844) as the array of fiducial elements (844) spans along the y-dimension. However, the length of the axis along the y- dimension of the elliptical shape for each fiducial element (844) gradually increases per each fiducial element (844) as the array of fiducial elements (844) spans along the y-dimension, such that fiducial elements (844) have a shortest axis along the y-dimension in the top side of optical fiducial region (840) and the longest axis along the y-dimension in the bottom side of optical fiducial region (840).

[00154] As best seen in FIG. 26, which shows the lower right-hand corner (842c) of optical fiducial region (840), the length of the axis along the y-dimension of the elliptical shape for each fiducial element (844) remains the same per each fiducial element (844) as the array of fiducial elements (844) spans along the x-dimension. However, the length of the axis along the x- dimension of the elliptical shape for each fiducial element (844) gradually decreases per each fiducial element (844) as the array of fiducial elements (844) spans along the x-dimension, such that fiducial elements (844) have a longest axis along the x-dimension in the left-hand side of optical fiducial region (840) and the shortest axis along the x-dimension in the right-hand side of optical fiducial region (840), as noted above with reference to FIG. 24.

[00155] In view of the foregoing, the elliptical shape of fiducial element (844) at upper left-hand corner (FIG. 23) has the longest length along the x-dimension and the shortest length along the y- dimension, the elliptical shape of fiducial element (844) at the upper right-hand corner (FIG. 24) has a circular shape representing the shortest axes along the x-dimension and y-dimension, the elliptical shape of fiducial element (844) at the lower left-hand corner (FIG. 25) has the longest axes along the x-dimension and y-dimension, and the elliptical shape of fiducial element (844) at the lower right-hand corner (FIG. 26) has the shortest length along the x- dimension and the longest length along the y-dimension. The elliptical shapes of the rest of fiducial elements (844) progressively transition from these extremes based on their positions along the x-dimension and y-dimension.

[09156] While objective lens assembly (140) and camera (142) may be primarily used to capture images of cells within imaging region (900) during use of system (100) in some examples, optical fiducial region (840) may nevertheless be within the field of view of objective lens assembly (140) and camera (142). Objective lens assembly (140) and camera (142) may thus capture an image of optical fiducial region (840) during a setup or calibration procedure. The image processing module (144) may analyze this image to determine whether cartridge (200) is appropriately positioned within the x-y plane relative to objective lens assembly (140) and camera (142). The configuration and arrangement of the unique elliptical shapes of fiducial elements (844) may facilitate image analysis by image processing module (144) to determine whether cartridge (200) is appropriately positioned within the x-y plane.

[00157] If the image analysis by image processing module (144) indicates that cartridge (200) is not appropriately positioned within the x-y plane, one or more adjustments may be made to ensure that cartridge (200) is appropriately positioned within the x-y plane. In some examples, cartridge (200) is secured to a mount that may be automatically moved by system (100) (e.g.. via one or more actuators) to move cartridge (200) into the appropriate position within the x-y plane. In some other examples, system (100) may notify an operator to reposition cartridge within the x-y plane. In still other examples, objective lens assembly (140) and camera (142) may be moved along an x-y plane to ensure appropriate alignment with cartridge. Such movement of objective lens assembly (140) and camera (142) may be manual or automated (e.g., performed by one or more actuators).

[00158] In addition to providing appropriate positioning of cartridge (200) relative to objective lens assembly (140) and camera (142) and/or other components of system (100) along an x-y plane, it may be beneficial to provide appropriate focusing of objective lens assembly (140) and camera (142) relative to cartridge (200) along the z-dimension. In some examples, such focusing may be performed as part of an initial setup or calibration process. In the present example, and as shown in FIG. 22, cartridge (200) includes a focusing feature (850) that may facilitate a focusing setup routine. Focusing feature (850) includes a plurality of focusing elements (852) that are positioned between optical fiducial region (840) and sampling channel (650). Focusing elements (852) are also within imaging region (900) in this example. It should therefore be understood that focusing feature (850) may be already within the field of view of objective lens assembly (140) and camera (142) when cartridge (200) is appropriately positioned within the x-y plane.

[00159] As shown in FIG. 27, each focusing element (852) of the present example includes a recess (854) in bottom surface (404) of layer (400). Each recess (854) has curved wall (856) such that the cross-sectional size of recess (854) along an x-y plane varies along the z-dimension. In some examples each recess (854) has a paraboloid shape, though other shapes may be used, including but not limited to recesses having a rectangular cross-sectional profile. Camera (142) may capture images of recess (854) at different positions along the z-dimension. Image processing module (144) may analyze these images to determine an appropriate focus along the z-dimension. For instance, image processing module (144) may determine the position of the bottom of recess (854) along the z-dimension and the position of the top of recess (854) along the z-dimension, and thereby identify the appropriate z-focus position (i.e., the position along the z-dimension at which to focus the image transmitted to camera (142)). In some examples, this appropriate z-focus position corresponds to the center of sampling channel (650) along the z-dimension. In some other examples, the appropriate z-focus position corresponds to a lower half of sampling channel (650) along the z-dimension or some other region of sampling channel (650) along the z-dimension.

[00169] In some examples, the position of cartridge (200) along the z-dimension may be adjusted in response to analysis of images focusing elements (852) captured by camera (142). In some examples, the position of a focusing lens (between objective lens assembly (140) and camera (142)) along the z-dimension and/or the position of camera (142) along the z-dimension may be adjusted in response to analysis of images focusing elements (852) captured by camera (142).

[00161] As another example of a focusing routine, referring to FIG. 28, camera (142) may be used to capture reference images of particles (850) conveyed along sampling channel (650) during an initial setup or calibration stage. Particles (850) may include cells, beads, or any other particles.

In the example shown, flow control fluid conveyed along fluidic channels (626) has provided horizontal focus to the fluid conveyed through fluidic channel (604), such that particles (850) are centered within sampling channel (650) along the y-dimension but are distributed across sampling channel (650) in the z-dimension within an upper vertical region (VR) and a lower vertical region (VR). Camera (142) may capture images of these particles (850) in this arrangement as particles (850) pass through imaging region (900). These images of particles (850) may be captured at different positions along the z-dimension.

[00162] By analyzing these images of particles (850), image processing module (144) may determine the vertical center position (Zo) of sampling channel (650) in the z-dimension. The appropriate z-focus position may then be established based on this determination. In some examples, this appropriate z-focus position corresponds to the vertical center position (Zo) of sampling channel (650) in the z-dimension. In some other examples, the appropriate z-focus position corresponds to a lower half of sampling channel (650) along the z-dimension or some other region of sampling channel (650) along the z-dimension. In examples where images of particles (850) rather than images of focusing elements ($52) are used to determine the appropriate z-focus position, the same kinds of adjustments may be made based on this determination as described above in the context of focusing elements (852).

[60163] L Example of Hybrid Fluidic Channels in Cartridge

[00164] As described above with reference to FIGS. 14A-14C, pneumatic valves (690, 692, 730, 732, 734, 736, 738, 740, 752, 754, 746, 748, 770, 774, 786, 790, 771, 773, 798, 791, 795) may be provided for fluidic channels (662, 664, 672, 710, 712, 714, 716, 718, 720) having cross- sectional profiles that are rectangular shaped. In some examples, pneumatic valves may be provided for fluidic channels having cross-sectional profiles that are rounded rather than rectangular. A valve al a fluidic channel having a rounded cross-sectional profile may require less pneumatic pressure in an overhead pneumatic channel to achieve full closure of the valve.

However, a fluidic channel having a square or rectangular cross-sectional profile may facilitate stable inertial flow focusing (e.g., along two focal planes in the z-dimension) more easily than a fluidic channel having a rounded cross-sectional profile. In addition, a fluidic channel having a square or rectangular cross-sectional profile may require less pressure to drive fluid flow through the fluidic channel. Thus, a “hybrid” cartridge may include fluidic channels that have regions with rounded cross-sectional profiles (e.g., where a valve would be positioned) and regions with rectangular cross-sectional profiles.

[00165] FIGS. 29-30 show an example of how cartridge (200) may be modified to include a hybrid fluidic channel configuration as described above. The cartridge of this example includes a sampling channel (2650) that is like sampling channel (650) of cartridge (200), such that sampling channel (2650) has a rectangular cross-sectional profile (along the y-z plane) along the entire length of sampling channel (2650). The features upstream of sampling channel (2650) may be the same as the features upstream of sampling channel (650). Sampling channel (2650) terminates in a junction (2660) that is like junction (660) of cartridge (2660). This junction (2660) further leads to a first outlet channel (2662) and a second outlet channel (2664).

The features downstream of first outlet channel (2662) may be the same as the features downstream of first outlet channel (662): and the features downstream of second outlet channel (2664) may be the same as the features downstream of second outlet channel (664).

[00166] First outlet channel (2662) of this example differs from first outlet channel (662) of cartridge (200) in that first outlet channel (662) has a portion (2663) having a rounded cross-sectional profile and an adjacent portion (2665) having a rectangular cross-sectional profile. Portion (2663) forms part of a valve (2692), which receives pneumatic pressure via a pneumatic channel (2322b). Pneumatic channel (2322b) is like pneumatic channel (322b). Portion (2665) extends downstream of portion (2663). Second outlet channel (2664) of this example differs from first outlet channel (662) of cartridge (200) in that first outlet channel (662) has a portion (2667) having a rounded cross-sectional profile and an adjacent portion (2669) having a rectangular cross-sectional profile. In some examples, the rounded cross-sectional profile of portion (2667) has a radius of curvature ranging from approximately 500 um to approximately 1000 um. Portion (2667) forms part of a valve (2690), which receives pneumatic pressure via a pneumatic channel (2322a). Pneumatic channel (2322a) is like pneumatic channel (322a).

Portion (2669) extends downstream of portion (2667).

[00167] It should be understood from FIG. 29 and the above description that pneumatic channel (2322b) is positioned over, and thus overlaps, portion (2663) along the z-dimension; but pneumatic channel (2322b) is not positioned over, and thus does not overlap, portion (2665) along the z- dimension. Similarly, pneumatic channel (23224) is positioned over, and thus overlaps, portion (2667) along the z-dimension; but pneumatic channel (232224) is not positioned over, and thus does not overlap, portion (2669) along the z-dimension.

[00168] FIGS. 30A-30B show an example of operation of valve (2690), though it should be understood that valve (2692) may operate in a similar fashion. As shown, pneumatic channel (2322a), which is in the form of a recess on the bottom surface of layer (2300), is positioned over portion

(2667), which is in the form of a recess on the bottom surface of layer (2400). Portion (2667) has a rounded cross-sectional profile along the y-z plane. In FIG. 30A, pneumatic channel (23224) is in a non-pressurized state, such that valve (2690) is in an open state. Fluid may thus flow freely through valve (2690) and thereby reach fluidic components via the remainder of second outlet channel (2664) downstream of valve (2690), including portion (2669). In FIG. 30B, pneumatic channel (2322a) is in a pressurized state. This pressurization in channel (23222) deforms the region of layer (2400) defining portion (2667) downwardly against the underlying surface (2502), thereby transitioning valve (2690) to a closed state. When pneumatic pressure is relieved in channel (2322a), the region of layer (2400) defining portion (2667) may resiliently return back to the state shown in FIG. 30A, thereby transitioning valve (2690) back to the closed state.

[00169] In some examples, the entire remainder of second outlet channel (2664) downstream of valve (2690) has a rectangular cross-sectional profile like portion (2669) described above. Also in some examples, the portion (2665) of first outlet channel (2602) having a rectangular cross- sectional profile may persist until reaching a secondary sorting region like secondary sorting region (204). Portions of fluidic channels along valves within such a secondary sorting region may also have a rounded cross-sectional profile like portions (2663, 2667) described above, with the remaining portions of such fluidic portions (e.g., portions that are not along valves) having rectangular cross-sectional profiles.

[00170] MI. Example of Another Cartridge

[00171] FIG. 31 shows an example of another cartridge (3000) that may be used with system (100) in place of cartridge (200). Except as otherwise noted below, cartridge (3000) of this example may be structurally configured and operable like cartridge (200). Cartridge (3000) of this example includes a sample fluid receiving region (3002) and a flow control fluid receiving region (3004). A flow channel (3006) leads from sample fluid receiving region (3002) to a junction (3006). A pair of flow channels (3008) lead from flow control fluid receiving region (3004) to junction (3006). A sampling channel (3012) extends from junction (3006) to a bend (3014), which leads to a sample output region (3018) via an outlet channel (3016).

[00172] A cell containing fluid may be communicated through sample fluid receiving region (3002), and a flow control fluid may be communicated through flow control fluid receiving region (3004). These two fluids meet at junction (3010), which may provide a flow focusing feature, with the merged fluids being conveyed along sampling channel (3012). Light source (130) and optical assembly (132) illuminate imaging region (3020); while objective lens assembly (140) and camera (142) capture images of cells within imaging region (3020). The fluid continues around bend (3014), through outlet channel (3016), and exits cartridge (3000) at sample output region (3018). The fluid that exits cartridge (3000) at sample output region (3018) may be further conveyed to a reservoir that is either integrated into the instrument of system (100) or is external to the instrument.

[00173] Cartridge (3000) is thus similar to cartridge (200) except that cartridge (3000) lacks the sorting capabilities of cartridge (200). Nevertheless, the other teachings provided above in the context of cartridge (200) may be readily applied to cartridge (3000).

[00174] IV. Examples of Combinations

[60175] The following examples relate to various non-exhaustive ways in which the teachings herein may be combined or applied. The following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, none of the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors.

If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.

[00176] Example 1

[09177] An apparatus comprising: a first fluidic path to receive a fluid containing particles; a first sorting region including: a second fluidic path, a third fluidic path, a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the first junction, a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction, and a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction, a second sorting region including: a fourth fluidic path, a fifth fluidic path, a second junction, the second fluidic path joining the fourth fluidic path and the fifth fluidic path at the second junction, the second junction being downstream of the first valve, a third valve to selectively permit or prevent communication of fluid along the fourth fluidic path downstream of the second junction, and a fourth valve to selectively permit or prevent communication of fluid along the fifth fluidic path downstream of the second junction; an outlet port to extract fluid from the third fluidic path, the outlet port being downstream of the second valve; a first well to receive fluid from the fourth fluidic path, the first well being downstream of the third valve; and a second well to receive fluid from the fifth fluidic path, the second well being downstream of the fourth valve.

[00178] Example 2

[00179] The apparatus of Example 1, the first valve including a pneumatic channel and a deformable layer, and the pneumatic channel to receive pneumatic pressure to deform the deformable layer.

[00180] Example 3

[00181] The apparatus of any of Examples 1 through 2, further comprising a camera to image particles conveyed along the first fluidic path.

[00182] Example 4

[00183] The apparatus of Example 3, further comprising a processor, the processor to classify particles from images captured by the camera.

[90184] Example 5

[00185] The apparatus of Example 4, the processor to control operation of the first valve, the second valve, the third valve, and the fourth valve.

[00186] Example 6

[00187] The apparatus of Example 5, the processor to select one or more valves from the first valve, the second valve, the third valve, and the fourth valve for activation in response to classification of particles.

[00188] Example 7

[00189] The apparatus of any of Examples 1 through 6, further comprising a cartridge, the first fluidic path, the second fluidic path, the third fluidic path, the fourth fluidic path, the fifth fluidic path, the first junction, and the second junction extending along the cartridge.

[00190] Example 8

[00191] The apparatus of Example 7, the first valve, the second valve, the third valve, and the fourth valve being in the cartridge.

[00192] Example 9

[00193] The apparatus of any of Examples 7 through 8, the outlet port providing a path for extraction of fluid from the cartridge via the fluidic path.

[00194] Example 10

[00195] The apparatus of any of Examples 7 through 9, the first well and the second well being in the cartridge.

[00196] Example 11

[00197] An apparatus comprising: a first layer including: a first surface, a second surface, a first fluidic path to receive a fluid containing particles, a second fluidic path, a third fluidic path, and a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the first junction; a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction; a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction; and a second layer including: a first surface, a second surface overlaying the first surface of the first layer, a first pneumatic channel to pneumatically pressurize the first valve and thereby transition the first valve to a closed state, a second pneumatic channel to pneumatically pressurize the second valve and thereby transition the first valve to a closed state, and a vent passageway, the vent passageway extending from the first surface to the second surface, the vent passageway being open to atmosphere.

[00198] Example 12

[00199] The apparatus of Example 11, the first fluidic path including a first recess in the first surface, the second fluidic path including a second recess in the first surface, and the third fluidic path including a third recess in the first surface.

[00200] Example 13

[60201] The apparatus of any of Examples 11 through 12, the first valve including a region of the second fluidic path adjacent to a corresponding region of the first pneumatic channel, and the second valve including a region of the third fluidic path adjacent to a corresponding region of the second pneumatic channel.

[00202] Example 14

[00203] The apparatus of Example 13, the first layer to deform in the region of the second fluidic path adjacent to the corresponding region of the first pneumatic channel in response to pneumatic pressurization of the first pneumatic channel, and the second layer to deform in the region of the third fluidic path adjacent to the corresponding region of the second pneumatic channel in response to pneumatic pressurization of the second pneumatic channel.

[00204] Example 15

[00205] The apparatus of any of Examples 11 through 14, the second layer further including one or more vent channels extending from the vent passageway along the second surface of the second layer, and the one or more vent channels to provide ventilation via the vent passageway.

[00206] Example 16

[00207] An apparatus comprising: a first layer including: a first surface, a second surface, a first fluidic path to receive a fluid containing particles, a second fluidic path; a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction; and a second layer including: a first surface, a second surface overlaying the first surface of the first layer, a first pneumatic channel to pneumatically pressurize the first valve and thereby transition the first valve to a closed state, a vent passageway, the vent passageway extending from the first surface to the second surface, the vent passageway being open to atmosphere, and a vent channel extending from the vent passageway along the first surface, the vent channel to provide ventilation via the vent passageway.

[00208] Example 17

[00209] The apparatus of Example 16, the first layer further including: a third fluidic path, and a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the tirst junction.

[00210] Example 18

[00211] The apparatus of Example 17, further comprising: a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction: and the second layer further including: a second pneumatic channel to pneumatically pressurize the second valve and thereby transition the first valve to a closed state.

[00212] Example 19

[00213] The apparatus of Example 18, the vent channel being positioned between the first pneumatic channel and the second pneumatic channel.

[00214] Example 20

[00215] The apparatus of any of clams 16 through 19, the first pneumatic channel including a first recess in the second surface of the second layer, the vent channel including a second recess in the second surface of the second layer, the first surface of the first layer defining a bottom of the first pneumatic channel, and the first surface of the first layer defining a bottom of the second vent channel.

[00216] Example 21

[00217] An apparatus comprising: a first fluidic path to receive a fluid containing particles; a first fluid input port to receive a first conduit to convey fluid containing particles to the first fluidic path; a flow control fluidic path to receive a flow control fluid to focus a flow of the fluid containing particles along the first fluidic path; a second fluid input port to receive a second conduit to convey flow control fluid to the flow control fluidic path; and a flush assembly including: a third fluid input port, a fluid output port, and a fluidic flush channel extending between the third fluid input port and the fluid output port, and the flush assembly to removably receive the first conduit and the second conduit, the flush assembly to permit a flush fluid to be communicated along a fluid path defined by a combination of the first conduit, the second conduit, and the fluidic tlush channel, such that the flush fluid enters the flush assembly via the third fluid input port and exits the flush assembly via the fluid output port.

[00218] Example 22

[00219] The apparatus of Example 21, further comprising a cartridge, the first fluidic path, the flow control fluidic path, and the fluidic flush channel extending along the cartridge.

[00220] Example 23

[00221] The apparatus of Example 22, the first fluid input port, the second fluid input port, the third fluid input port, and the fluid output port extending into the cartridge.

[00222] Example 24

[00223] The apparatus of any of Examples 21 through 23, the first conduit including a first flexible tube, and the second conduit including a second flexible tube.

[00224] Example 25

[00225] A method comprising: inserting a first conduit into a first fluid input port of a cartridge; inserting a second conduit into a second fluid input of the cartridge; conveying a fluid containing particles along a first fluidic path of the cartridge via the first conduit and the first fluid input port; conveying a second fluid along a second fluidic path of the cartridge via the second conduit and the second fluid input port; removing the first conduit from the first fluid input port; removing the second conduit from the second fluid input port; inserting the first conduit into a third port of the cartridge; inserting the second conduit into a fourth port of the cartridge, the third port and the fourth port being coupled via a first fluidic flush channel to form a first flush assembly; and communicating a flushing fluid through the first conduit and the second conduit and further through the first flush assembly.

[00226] Example 26

[00227] The method of Example 25, further comprising capturing images of particles conveyed along the first fluidic path.

[00228] Example 27

[00229] The method of any of Examples 25 through 26, the second fluid providing a focus to flow of the fluid containing particles along the first fluidic path.

[00230] Example 28

[00231] The method of any of Examples 25 through 27, the third port defining an output port of the first flush assembly, the fourth port defining an input port of the first flush assembly, communicating a flushing fluid through the first conduit and the second conduit and further through the first flush assembly including: communicating the flush fluid into the flush assembly via the first conduit, and communicating the flush fluid out of the flush assembly via the second conduit.

[00232] Example 29

[00233] The method of any of Examples 25 through 28, further comprising: inserting a fourth conduit into a fifth fluid input port of the cartridge; inserting a fifth conduit into a sixth fluid input port of the cartridge; conveying the fluid containing particles from the first fluidic path to a receptacle via the fourth conduit and the fifth fluid input port; and conveying a third fluid along a third fluidic path of the cartridge via the fifth conduit and the sixth fluid input port.

[00234] Example 30

[00235] The method of Example 29, further comprising: removing the fourth conduit from the fifth fluid input port; removing the fifth conduit from the sixth fluid input port; inserting the fourth conduit into a seventh port of the cartridge; inserting the fifth conduit into a eighth port of the cartridge, the seventh port and the eighth port being coupled via a second fluidic flush channel to form a second first flush assembly; and communicating a flushing fluid through the fourth conduit and the fifth conduit and further through the second flush assembly.

[00236] Example 31 [0602371 An apparatus comprising: a cartridge, the cartridge including: a first fluidic path to receive a fluid containing particles, the first fluidic path providing an imaging region to capture images of particles conveyed along the first fluidic path, and an optical fiducial region including a plurality of fiducial elements, each fiducial element of the plurality of fiducial elements having a shape that differs from shapes of the other fiducial elements of the plurality of fiducial elements; and a camera having a field of view, the cartridge being positionable to provide the imaging region and the optical fiducial region simultaneously within the field of view.

[00238] Example 32

[09239] The apparatus of Example 31, the plurality of fiducial elements having elliptical shapes, each elliptical shape having a major axis and a minor axis, major axes and minor axes varying among the plurality of fiducial elements.

[00240] Example 33

[00241] The apparatus of any of Examples 31 through 32, further comprising a processor, the processor to process an image of the optical fiducial region captured by the camera, to thereby determine whether the cartridge is aligned with the camera.

[00242] Example 34

[00243] The apparatus of Example 33, the processor to further process an image of a particle in the fluidic path captured by the camera, to thereby classify the particle.

[90244] Example 35

[00245] The apparatus of any of Examples 33 through 34, the camera to capture images of particles in the fluidic path at different positions along a height of the fluidic path, and the processor to process the images of particles in the fluidic path at different positions along the height of the fluidic path to thereby determine a focus location.

[00246] Example 36

[00247] The apparatus of Example 35, the height of the fluidic path having a vertical center position, the processor to determine the focus location at the vertical center position.

[00248] Example 37

[00249] The apparatus of Example 35, the height of the fluidic path having a vertical center position, the processor to determine the focus location at a vertical position below the vertical center position.

[00250] Example 38

[00251] The apparatus of any of Examples 35 through 37, the particles including beads.

[00252] Example 39

[00253] The apparatus of any of Examples 31 through 38, the cartridge further including a layer having a first surface and a second surface, and the plurality of fiducial elements including recesses in the first surface.

[00254] Example 40

[00255] The apparatus of any of Examples 31 through 39, the cartridge further including a focusing region having a focusing feature, the focusing feature, the cartridge being positionable to provide the imaging region, the optical fiducial region, and the focusing feature simultaneously within the field of view, and the camera having an optical axis extending along a first dimension, the focusing feature having a depth along the first dimension.

[00256] Example 41

[00257] The apparatus of Example 40, the cartridge further including a layer having a first surface and a second surface, and the focusing feature including a recess in the first surface.

[90258] Example 42

[00259] The apparatus of Example 41, the focusing feature having a curved wall, the curved wall providing a cross-sectional area along a first plane that varies along the first dimension, and the first plane being perpendicular to the first dimension.

[09269] Example 43

[00261] The apparatus of any of Examples 31 through 42, further comprising a processor, the camera to capture images of the focusing feature at different positions along a height of the focusing feature, and the processor to process the images of the focusing feature at different positions along the height of the focusing feature to thereby determine a focus location.

[00262] Example 44

[00263] An apparatus comprising: a first layer; a second layer positioned over the first layer, the second layer including: a first fluidic channel, the first fluidic channel including a first portion and a second portion adjacent to the first portion, the first portion having a rectangular cross-section, the second portion having a rounded cross-section; and a third layer positioned over the second layer, the third layer including a first pneumatic channel, the first pneumatic channel to apply pneumatic pressure against the second portion of the first fluidic channel to thereby deform the second portion against the first layer and thereby prevent communication of fluid through the first fluidic channel.

[90264] Example 45

[00265] The apparatus of Example 44, the second layer including resilient material to return the second portion to a non-deformed state in response to removal of pneumatic pressure in the first pneumatic channel.

[00266] Example 46

[00267] The apparatus of any of Examples 44 through 45, the first fluidic channel to convey a fluid containing particles.

[90268] Example 47

[00269] The apparatus of any of Examples 44 through 46, the second layer having a first surface adjacent to the first layer and a second surface adjacent to the third layer, and the first fluidic channel including a recess in the first surface of the second layer.

[00270] Example 48

[60271] The apparatus of any of Examples 44 through 47, the third layer having a first surface adjacent to the second layer and a second surface, and the first pneumatic channel including a recess in the first surface of the third layer.

[00272] Example 49

[00273] The apparatus of any of Examples 44 through 48, the first pneumatic channel being positioned over the second portion such that the first pneumatic channel overlaps the second portion.

[00274] Example 50

[00275] The apparatus of any of Examples 44 through 49, the first portion being positioned such that the first pneumatic channel does not overlap the first portion.

[90276] Example 51 [0602771 The apparatus of any of Examples 44 through 50, the first layer further including: a junction upstream of the first fluidic channel, a second fluidic channel upstream of the junction, and a third fluidic channel downstream of the junction.

[00278] Example 52

[060279] The apparatus of Example 51, the third fluidic channel including a third portion and a fourth portion adjacent to the third portion, the third portion having a rectangular cross-section, and the fourth portion having a rounded cross-section.

[00280] Example 53

[00281] The apparatus of Example 52, the third layer further including a second pneumatic channel, the second pneumatic channel to apply pneumatic pressure against the fourth portion of the third fluidic channel to thereby deform the fourth portion against the first layer and thereby prevent communication of fluid through the third fluidic channel.

[00282] V. Miscellaneous

[00283] While the examples provided above include cells or beads as particles, the teachings herein may be readily applied to other contexts where other kinds of particles are used in addition to or in lieu of cells or beads.

[00284] The foregoing description is provided to enable a person skilled in the art to practice the various configurations described herein. While the subject technology has been particularly described with reference to the various figures and configurations, it should be understood that these are for illustration purposes only and should not be taken as limiting the scope of the subject technology. The subject matter described herein is not limited in its application to the details of construction and the arrangement of components set forth in the description herein or illustrated in the drawings hereof. The subject matter described herein is capable of other implementations and of being practiced or of being carried out in various ways.

[00285] It is to be understood that the above description is intended to be illustrative, and not restrictive.

For example, the above-described examples (and/or aspects thereof) may be used in combination with each other. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the presently described subject matter without departing from its scope. While the dimensions, types of materials and coatings described herein are intended to define the parameters of the disclosed subject matter, they are by no means limiting and instead illustrations. Many further examples will be apparent to those of skill in the art upon reviewing the above description. The scope of the disclosed subject matter should, therefore, be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. Further, the limitations of the following claims are not written in means—plus-function format and are not intended to be interpreted based on 35 U.S.C. §112(f), unless and until such claim limitations expressly use the phrase “means for” followed by a statement of function void of further structure.

[00286] The following claims recite aspects of certain examples of the disclosed subject matter and are considered to be part of the above disclosure. These aspects may be combined with one another.

[00287] The disclosure further comprises the following clauses, which correspond to the appended

Dutch-language claims:

CLAUSES

I. An apparatus comprising: a first fluidic path to receive a fluid containing particles; a first sorting region including: a second fluidic path, a third fluidic path,

a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the first junction,

a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction, and a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction, a second sorting region including:

a fourth fluidic path,

a fifth fluidic path,

a second junction, the second fluidic path joining the fourth fluidic path and the fifth fluidic path at the second junction, the second junction being downstream of the first valve,

a third valve to selectively permit or prevent communication of fluid along the fourth fluidic path downstream of the second junction, and a fourth valve to selectively permit or prevent communication of fluid along the fifth fluidic path downstream of the second junction;

an outlet port to extract fluid from the third fluidic path, the outlet port being downstream of the second valve; a first well to receive fluid from the fourth fluidic path, the first well being downstream of the third valve; and a second well to receive fluid from the fifth fluidic path, the second well being downstream of the fourth valve. 2. The apparatus of clause 1, the first valve including a pneumatic channel and a deformable layer, and the pneumatic channel to receive pneumatic pressure to deform the deformable layer.

3. The apparatus of any of clauses 1 through 2, further comprising a camera to image particles conveyed along the first fluidic path.

4. The apparatus of clause 3, further comprising a processor, the processor to classify particles from images captured by the camera. 5. The apparatus of clause 4, the processor to control operation of the first valve, the second valve, the third valve, and the fourth valve.

6. The apparatus of clause 5, the processor to select one or more valves from the first valve, the second valve, the third valve, and the fourth valve for activation in response to classification of particles.

7. The apparatus of any of clauses 1 through 6, further comprising a cartridge, the first fluidic path, the second fluidic path, the third fluidic path, the fourth tluidic path, the fifth fluidic path, the first junction, and the second junction extending along the cartridge. 8. The apparatus of clause 7, the first valve, the second valve, the third valve, and the fourth valve being in the cartridge. 9. The apparatus of any of clauses 7 through 8, the outlet port providing a path for extraction of fluid from the cartridge via the fluidic path.

10. The apparatus of any of clauses 7 through 9, the first well and the second well being in the cartridge. it.

An apparatus comprising: a first layer including: a first surface, a second surface, a first fluidic path to receive a fluid containing particles, a second fluidic path, a third fluidic path, and a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the first junction; a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction; a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction; and a second layer including: a first surface, a second surface overlaying the first surface of the first layer,

a first pneumatic channel to pneumatically pressurize the first valve and thereby transition the first valve to a closed state, a second pneumatic channel to pneumatically pressurize the second valve and thereby transition the first valve to a closed state, and a vent passageway, the vent passageway extending from the first surface to the second surface, the vent passageway being open to atmosphere.

12. The apparatus of clause 11, the first fluidic path including a first recess in the first surface, the second fluidic path including a second recess in the first surface, and the third fluidic path including a third recess in the first surface.

13. The apparatus of any of clauses 11 through 12, the first valve including a region of the second fluidic path adjacent to a corresponding region of the first pneumatic channel, and the second valve including a region of the third fluidic path adjacent to a corresponding region of the second pneumatic channel.

14. The apparatus of clause 13, the first layer to deform in the region of the second fluidic path adjacent to the corresponding region of the first pneumatic channel in response to pneumatic pressurization of the first pneumatic channel, and the second layer to deform in the region of the third fluidic path adjacent to the corresponding region of the second pneumatic channel in response to pneumatic pressurization of the second pneumatic channel.

15. The apparatus of any of clauses 11 through 14, the second layer further including one or more vent channels extending from the vent passageway along the second surface of the second layer, and the one or more vent channels to provide ventilation via the vent passageway. 16. An apparatus comprising: a first layer including: a first surface, a second surface, a first fluidic path to receive a fluid containing particles, and a second fluidic path; a first valve to selectively permit or prevent communication of fluid along the second fluidic path downstream of the first junction; and a second layer including: a first surface, a second surface overlaying the first surface of the first layer, a first pneumatic channel to pneumatically pressurize the first valve and thereby transition the first valve to a closed state, a vent passageway, the vent passageway extending from the first surface to the second surface, the vent passageway being open to atmosphere, and a vent channel extending from the vent passageway along the first surface, the vent channel to provide ventilation via the vent passageway.

17. The apparatus of clause 16, the first layer further including: a third fluidic path, and a first junction, the first fluidic path joining the second fluidic path and the third fluidic path at the first junction.

18. The apparatus of clause 17, further comprising: a second valve to selectively permit or prevent communication of fluid along the third fluidic path downstream of the first junction; and the second layer further including: a second pneumatic channel to pneumatically pressurize the second valve and thereby transition the first valve to a closed state. 19. The apparatus of clause 18, the vent channel being positioned between the first pneumatic channel and the second pneumatic channel.

20. The apparatus of any of clams 16 through 19, the first pneumatic channel including a first recess in the second surface of the second layer, the vent channel including a second recess in the second surface of the second layer, the first surface of the first layer defining a bottom of the first pneumatic channel, and the first surface of the first layer defining a bottom of the second vent channel.

21. An apparatus comprising: a first fluidic path to receive a fluid containing particles; a first fluid input port to receive a first conduit to convey fluid containing particles to the first fluidic path;

a flow control fluidic path to receive a flow control fluid to focus a flow of the fluid containing particles along the first fluidic path; a second fluid input port to receive a second conduit to convey flow control fluid to the flow control fluidic path; and a flush assembly including: a third fluid input port, a fluid output port, and a fluidic flush channel extending between the third fluid input port and the fluid output port, the flush assembly to removably receive the first conduit and the second conduit, and the flush assembly to permit a flush fluid to be communicated along a fluid path defined by a combination of the first conduit, the second conduit, and the fluidic flush channel, such that the flush fluid enters the flush assembly via the third fluid input port and exits the flush assembly via the fluid output port. 22. The apparatus of clause 21, further comprising a cartridge, the first fluidic path, the flow control fluidic path, and the fluidic flush channel extending along the cartridge.

23. The apparatus of clause 22, the first fluid input port, the second fluid input port, the third fluid input port, and the fluid output port extending into the cartridge. 24. The apparatus of any of clauses 21 through 23, the first conduit including a first flexible tube, and the second conduit including a second flexible tube. 25. A method comprising: inserting a first conduit into a first fluid input port of a cartridge; inserting a second conduit into a second fluid input of the cartridge; conveying a fluid containing particles along a first fluidic path of the cartridge via the first conduit and the first fluid input port; conveying a second fluid along a second fluidic path of the cartridge via the second conduit and the second fluid input port; removing the first conduit from the first fluid input port;

removing the second conduit from the second fluid input port;

inserting the first conduit into a third port of the cartridge;

inserting the second conduit into a fourth port of the cartridge, the third port and the fourth port being coupled via a first fluidic flush channel to form a first flush assembly; and communicating a flushing fluid through the first conduit and the second conduit and further through the first flush assembly. 26. The method of clause 25, further comprising capturing images of particles conveyed along the first fluidic path.

27. The method of any of clauses 25 through 26, the second fluid providing a focus to flow of the fluid containing particles along the first fluidic path.

28. The method of any of clauses 25 through 27, the third port defining an output port of the first flush assembly. the fourth port defining an input port of the first flush assembly, communicating a flushing fluid through the first conduit and the second conduit and further through the first flush assembly including:

communicating the flush fluid into the flush assembly via the first conduit, and communicating the flush fluid out of the flush assembly via the second conduit. 29. The method of any of clauses 25 through 28, further comprising: inserting a fourth conduit into a fifth fluid input port of the cartridge; inserting a fifth conduit into a sixth fluid input port of the cartridge; conveying the fluid containing particles from the first fluidic path to a receptacle via the fourth conduit and the fifth fluid input port; and conveying a third fluid along a third fluidic path of the cartridge via the fifth conduit and the sixth fluid input port. 30. The method of clause 29, further comprising: removing the fourth conduit from the fifth fluid input port; removing the fifth conduit from the sixth fluid input port; inserting the fourth conduit into a seventh port of the cartridge;

inserting the fifth conduit into a eighth port of the cartridge, the seventh port and the eighth port being coupled via a second fluidic flush channel to form a second first flush assembly; and communicating a flushing fluid through the fourth conduit and the fifth conduit and further through the second flush assembly. 31. An apparatus comprising: a cartridge, the cartridge including: a first fluidic path to receive a fluid containing particles, the first fluidic path providing an imaging region to capture images of particles conveyed along the first fluidic path, and an optical fiducial region including a plurality of fiducial elements, each fiducial element of the plurality of fiducial elements having a shape that differs from shapes of the other fiducial elements of the plurality of fiducial elements; and a camera having a field of view, the cartridge being positionable to provide the imaging region and the optical fiducial region simultaneously within the field of view. 32. The apparatus of clause 31, the plurality of fiducial elements having elliptical shapes,

each elliptical shape having a major axis and a minor axis, major axes and minor axes varying among the plurality of fiducial elements.

33. The apparatus of any of clauses 31 through 32, further comprising a processor, the processor to process an image of the optical fiducial region captured by the camera, to thereby determine whether the cartridge is aligned with the camera.

34. The apparatus of clause 33, the processor to further process an image of a particle in the fluidic path captured by the camera, to thereby classify the particle.

35. The apparatus of any of clauses 33 through 34, the camera to capture images of particles in the fluidic path at different positions along a height of the fluidic path, and the processor to process the images of particles in the fluidic path at different positions along the height of the fluidic path to thereby determine a focus location.

36. The apparatus of clause 35, the height of the fluidic path having a vertical center position, the processor to determine the focus location at the vertical center position. 37. The apparatus of clause 35, the height of the fluidic path having a vertical center position,

S the processor to determine the focus location at a vertical position below the vertical center position. 38. The apparatus of any of clauses 35 through 37, the particles including beads. 39. The apparatus of any of clauses 31 through 38, the cartridge further including a layer having a first surface and a second surface, and the plurality of fiducial elements including recesses in the first surface. 40. The apparatus of any of clauses 31 through 39, the cartridge further including a focusing region having a focusing feature, the focusing feature, the cartridge being positionable to provide the imaging region, the optical fiducial region, and the focusing feature simultaneously within the field of view, and the camera having an optical axis extending along a first dimension, the focusing feature having a depth along the first dimension. 41. The apparatus of clause 40, the cartridge further including a layer having a first surface and a second surface, and the focusing feature including a recess in the first surface. 42. The apparatus of clause 41, the focusing feature having a curved wall, the curved wall providing a cross-sectional area along a first plane that varies along the first dimension, and the first plane being perpendicular to the first dimension. 43. The apparatus of any of clauses 31 through 42, further comprising a processor, the camera to capture images of the focusing feature at different positions along a height of the focusing feature, and the processor to process the images of the focusing feature at different positions along the height of the focusing feature to thereby determine a focus location. 44. An apparatus comprising: a first layer; a second layer positioned over the first layer, the second layer including:

a first fluidic channel, the first fluidic channel including a first portion and a second portion adjacent to the first portion, the first portion having a rectangular cross-section, the second portion having a rounded cross- section; and a third layer positioned over the second layer, the third layer including a first pneumatic channel, the first pneumatic channel to apply pneumatic pressure against the second portion of the first fluidic channel to thereby deform the second portion against the first layer and thereby prevent communication of fluid through the first fluidic channel.

45. The apparatus of clause 44, the second layer including resilient material to return the second portion to a non-deformed state in response to removal of pneumatic pressure in the first pneumatic channel.

46. The apparatus of any of clauses 44 through 45, the first fluidic channel to convey a fluid containing particles.

47. The apparatus of any of clauses 44 through 46, the second layer having a first surface adjacent to the first layer and a second surface adjacent to the third layer, and the first fluidic channel including a recess in the first surface of the second layer.

48. The apparatus of any of clauses 44 through 47, the third layer having a first surface adjacent to the second layer and a second surface, and the first pneumatic channel including a recess in the first surface of the third layer.

49. The apparatus of any of clauses 44 through 48, the first pneumatic channel being positioned over the second portion such that the first pneumatic channel overlaps the second portion.

50. The apparatus of any of clauses 44 through 49, the first portion being positioned such that the first pneumatic channel does not overlap the first portion. 51. The apparatus of any of clauses 44 through 50, the first layer further including: a junction upstream of the first fluidic channel, a second fluidic channel upstream of the junction, and a third fluidic channel downstream of the junction, 52. The apparatus of clause 51, the third fluidic channel including a third portion and a fourth portion adjacent to the third portion, the third portion having a rectangular cross-section, and the fourth

S portion having a rounded cross-section. 53. The apparatus of clause 52, the third layer further including a second pneumatic channel, the second pneumatic channel to apply pneumatic pressure against the fourth portion of the third fluidic channel to thereby deform the fourth portion against the first layer and thereby prevent communication of fluid through the third fluidic channel.

Claims

CONCLUSIONS

1. An apparatus comprising: a first fluid path for receiving a fluid comprising particles; a first sorting region including: a second fluid path, a third fluid path, a first connection, the first fluid path connecting to the second fluid path and the third fluid path at the first connection, a first valve for selectively allowing or preventing fluid communication along the second fluid path downstream of the first connection, and a second valve for selectively allowing or preventing fluid communication along the third fluid path downstream of the first connection, a second sorting region including: a fourth fluid path, a fifth fluid path, a second connection, the second fluid path connecting to the fourth fluid path and the fifth fluid path at the second connection, the second connection being downstream of the first valve, a third valve for selectively allowing or preventing fluid communication along the fourth fluid path downstream of the second connection, and a fourth valve for selectively allowing or preventing fluid communication along the fifth fluid path downstream of the second connection; an outlet port for withdrawing fluid from the third fluid path, the outlet port being located downstream of the second valve; a first well for receiving fluid from the fourth fluid path, the first well being located downstream of the third valve; and a second well for receiving fluid from the fifth fluid path, the second well being located downstream of the fourth valve.

2. The apparatus of claim 1, wherein the first valve comprises a pneumatic channel and a deformable layer, and the pneumatic channel is adapted to receive pneumatic pressure to deform the deformable layer.

3. The apparatus of any one of claims 1 to 2, further comprising a camera for imaging particles transported along the first fluid path.

4. The apparatus of claim 3, further comprising a processor, the processor classifying particles from images captured by the camera.

5. The apparatus of claim 4, wherein the processor controls the operation of the first valve, the second valve, the third valve and the fourth valve.

6. The apparatus of claim 5, wherein the processor selects one or more valves from the first valve, the second valve, the third valve and the fourth valve for activation in response to the classification of particles.

7. The apparatus of any one of claims | to 6, further comprising a cartridge, the first fluid path, the second fluid path, the third fluid path, the fourth fluid path, the fifth fluid path, the first node and the second node extending along the cartridge.

8. The apparatus of claim 7, wherein the first valve, the second valve, the third valve and the fourth valve are located in the cartridge.

9. The apparatus of any one of claims 7 to 8, wherein the outlet port provides a path for extracting fluid from the cartridge via the fluid path.

10. Apparatus according to any one of claims 7 to 9, wherein the first well and the second well are located in the cartridge.

11. An apparatus comprising: a first layer including: a first surface, a second surface, a first fluid path for receiving a fluid comprising particles, a second fluid path, a third fluid path, and a first connection, the first fluid path connecting to the second fluid path and the third fluid path at the first connection; a first valve for selectively allowing or preventing fluid communication along the second fluid path downstream of the first connection; a second valve for selectively allowing or preventing fluid communication along the third fluid path downstream of the first connection; and a second layer including: a first surface, a second surface overlying the first surface of the first layer, a first pneumatic conduit for pneumatically pressurizing the first valve and thereby transferring the first valve to a closed state, a second pneumatic conduit for pneumatically pressurizing the second valve and thereby transferring the first valve to a closed state, and a ventilation passage, the ventilation passage extending from the first surface to the second surface, the ventilation passage being open to the atmosphere.

12. The apparatus of claim 11, wherein the first fluid path comprises a first recess in the first surface, the second fluid path comprises a second recess in the first surface, and the third fluid path comprises a third recess in the first surface.

The apparatus of any one of claims 11 to 12, wherein the first valve comprises a region of the second fluid path adjacent to a corresponding region of the first pneumatic channel, and the second valve comprises a region of the third fluid path adjacent to a corresponding region of the second pneumatic channel.

14. The apparatus of claim 13, wherein the first layer deforms in the region of the second fluid path adjacent to the corresponding region of the first pneumatic channel in response to pneumatic pressurization of the first pneumatic channel, and the second layer deforms in the region of the third fluid path adjacent to the corresponding region of the second pneumatic channel in response to pneumatic pressurization of the second pneumatic channel.

15. The apparatus of any of claims 11 to 14, wherein the second layer further comprises one or more ventilation channels extending from the ventilation passage along the second surface of the second layer, and the one or more ventilation channels to provide ventilation through the ventilation passage.

16. An apparatus comprising: a first layer including: a first surface, a second surface, a first fluid path for receiving a fluid comprising particles, and a second fluid path; a first valve for selectively allowing or preventing fluid communication along the second fluid path downstream of the first connection; and a second layer including: a first surface, a second surface overlying the first surface of the first layer, a first pneumatic channel for pneumatically pressurizing the first valve and thereby transferring the first valve to a closed state, a ventilation passageway, the ventilation passageway extending from the first surface to the second surface, the ventilation passageway being open to the atmosphere, and a ventilation channel extending from the ventilation passageway along the first surface, the ventilation channel providing ventilation through the ventilation passageway.

17. The apparatus of claim 16, wherein the first layer further comprises: a third fluid path, and a first connection, the first fluid path connecting to the second fluid path and the third fluid path connecting to the first connection.

18. The apparatus of claim 17, further comprising: a second valve for selectively allowing or preventing fluid communication along the third fluid path downstream of the first connection; and the second layer further comprises: a second pneumatic conduit for pneumatically pressurizing the second valve and thereby transferring the first valve to a closed state.

19. The apparatus of claim 18, wherein the ventilation duct is positioned between the first pneumatic duct and the second pneumatic duct.

20. The apparatus of any one of claims 16 to 19, wherein the first pneumatic channel comprises a first recess in the second surface of the second layer, the ventilation channel comprises a second recess in the second surface of the second layer, the first surface of the first layer defines a bottom of the first pneumatic channel, and the first surface of the first layer defines a bottom of the second ventilation channel.

21. An apparatus comprising: a first fluid path for receiving a fluid comprising particles; a first fluid inlet port for receiving a first conduit for conveying fluid comprising particles to the first fluid path; a flow control fluid path for receiving a flow control fluid for focusing a flow of the fluid comprising particles along the first fluid path; a second fluid inlet port for receiving a second conduit for conveying flow control fluid to the flow control fluid path; and a recessed assembly including: a third fluid inlet port, a fluid outlet port, and a fluid flushing channel extending between the third fluid inlet port and the fluid outlet port, the recessed assembly for removably receiving the first conduit and the second conduit, and the flushing assembly for permitting a flushing fluid to be communicated along a fluid path defined by a combination of the first conduit, the second conduit, and the fluid flushing channel such that the flushing fluid enters the flushing assembly through the third fluid inlet port and exits the flushing assembly through the fluid outlet port.

22. The apparatus of claim 21 further comprising a cartridge, the first fluid path, the flow control fluid path, and the fluid flushing channel extending along the cartridge.

23. The apparatus of claim 22, wherein the first fluid input port, the second fluid input port, the third fluid input port and the fluid output port extend into the cartridge.

24. The apparatus of any one of claims 21 to 23, wherein the first conduit comprises a first flexible tube, and the second conduit comprises a second flexible tube.

25. A method comprising: inserting a first conduit into a first fluid input port of a cartridge; inserting a second conduit into a second fluid input port of the cartridge; conveying a fluid comprising particles along a first fluid path of the cartridge via the first conduit and the first fluid input port; conveying a second fluid along a second fluid path of the cartridge via the second conduit and the second fluid input port; removing the first conduit from the first fluid input port: removing the second conduit from the second fluid input port; inserting the first conduit into a third port of the cartridge; inserting the second conduit into a fourth port of the cartridge, the third port and the fourth port being coupled via a first fluid flush channel to form a first flush assembly; and communicating a flush fluid through the first channel and the second channel and further through the first flush assembly.

26. The method of claim 25 further comprising capturing images of particles transported along the first fluid path.

27. A method according to any one of claims 25 to 26, wherein the second fluid provides a focus for the flow of the fluid comprising particles along the first fluid path.

28. The method of any of claims 25 to 27, wherein the third port defines an exit port of the first coil assembly, and the fourth port defines an entrance port of the first coil assembly, wherein a flushing fluid is communicated through the first conduit and the second conduit and further through the first coil assembly including: communicating the flushing fluid into the coil assembly via the first conduit, and communicating the flushing fluid from the coil assembly via the second conduit.

29. The method of any one of claims 25 to 28, further comprising: inserting a fourth conduit into a fifth fluid input port of the cartridge; inserting a fifth conduit into a sixth fluid input port of the cartridge; conveying the fluid comprising particles from the first fluid path to a receptacle via the fourth conduit and the fifth fluid input port; and conveying a third fluid along a third fluid path of the cartridge via the fifth conduit and the sixth fluid input port.

30. The method of claim 29 further comprising: removing the fourth conduit from the fifth fluid input port; removing the fifth conduit from the sixth fluid input port; inserting the fourth conduit into a seventh port of the cartridge; inserting the fifth conduit into an eighth port of the cartridge, the seventh port and the eighth port being coupled via a second fluid flush channel to form a second first flush assembly; and communicating a flush fluid through the fourth conduit and the fifth conduit and further through the second flush assembly.

31. An apparatus comprising: a cartridge, the cartridge comprising: a first fluid path for receiving a fluid comprising particles, the first fluid path providing an imaging region for capturing images of particles transported along the first fluid path, and an optical reference region comprising a plurality of reference elements, each reference element of the plurality of reference elements having a shape different from the shapes of the other reference elements of the plurality of reference elements; and a camera having a field of view, the cartridge being positionable to provide the imaging region and the optical reference region simultaneously within the field of view.

32. The apparatus of claim 31, wherein the plurality of fixed elements have elliptical shapes, each elliptical shape having a major axis and a minor axis, the major axes and minor axes varying among the plurality of fixed elements.

33. The apparatus of any one of claims 31 to 32, further comprising a processor, the processor processing an image of the optical reference area captured by the camera to thereby determine whether the cartridge is aligned with the camera.

34. The apparatus of claim 33, wherein the processor further processes an image of a particle in the fluid path captured by the camera to thereby classify the particle.

35. The apparatus of any one of claims 33 to 34, wherein the camera captures images of particles in the fluid path at different positions along a height of the fluid path, and the processor processes the images of particles in the fluid path at different positions along the height of the fluid path to thereby determine a focus location,

36. The apparatus of claim 35, wherein the height of the fluid path has a vertical center position, the processor determining the focus location at the vertical center position.

37. The apparatus of claim 35, wherein the height of the fluid path has a vertical center position, the processor determining the focus location at a vertical position below the vertical center position.

38. The device of any one of claims 35 to 37, wherein the particles comprise beads.

39. The device of any one of claims 31 to 38, wherein the cartridge further comprises a layer having a first surface and a second surface, and the plurality of reference elements including recesses in the first surface.

40. The apparatus of any one of claims 31 to 39, wherein the cartridge further comprises a focusing region having a focusing means, the focusing means being positionable to simultaneously provide the imaging region, the optical reference region and the focusing means within the field of view, and wherein the camera has an optical axis extending along a first dimension, the focusing means having a depth along the first dimension.

41. The apparatus of claim 40, wherein the cartridge further comprises a layer having a first surface and a second surface, and the focusing means encloses a recess in the first surface.

42. The apparatus of claim 41, wherein the focusing means has a curved wall, the curved wall providing a cross-sectional area along a first plane that varies along the first dimension, and the first plane is perpendicular to the first dimension.

43. The apparatus of any one of claims 31 to 42, further comprising a processor, the camera to capture images of the focusing means at different positions along a height of the focusing means, and the processor to process the images of the focusing means at different positions along the height of the focusing means to thereby determine a focus location.

44. An apparatus comprising: a first layer; a second layer positioned over the first layer, the second layer comprising: a first fluid channel, the first fluid channel comprising a first portion and a second portion adjacent to the first portion, the first portion having a rectangular cross-section, and the second portion having a rounded cross-section; and a third layer positioned over the second layer, the third layer comprising a first pneumatic channel, the first pneumatic channel applying pneumatic pressure to the second portion of the first fluid channel to thereby deform the second portion against the first layer and thereby prevent communication of fluid through the first fluid channel.

45. The apparatus of claim 44, wherein the second layer comprises resilient material for returning the second portion to an undeformed state in response to removal of pneumatic pressure in the first pneumatic channel.

46. The apparatus of any one of claims 44 to 45, wherein the first fluid channel transports a fluid containing particles.

47. The apparatus of any one of claims 44 to 46, wherein the second layer has a first surface adjacent to the first layer and a second surface adjacent to the third layer, and the first fluid channel comprises a recess in the first surface of the second layer,

48. The apparatus of any of claims 44 to 47, wherein the third layer has a first surface adjacent to the second layer and a second surface, and the first pneumatic channel comprises a recess in the first surface of the third layer.

49. The apparatus of any one of claims 44 to 48, wherein the first pneumatic channel is positioned over the second portion such that the first pneumatic channel overlaps the second portion.

50. The apparatus of any one of claims 44 to 49, wherein the first portion is positioned such that the first pneumatic channel does not overlap the first portion.

51. The apparatus of any of claims 44 to 50, wherein the first layer further comprises: a junction upstream of the first fluid channel, a second fluid channel upstream of the junction, and a third fluid channel downstream of the junction.

52. The apparatus of claim 51, wherein the third fluid channel comprises a third portion and a fourth portion adjacent to the third portion, the third portion having a rectangular cross-section, and the fourth portion having a rounded cross-section.

53. The apparatus of claim 52, wherein the third layer further comprises a second pneumatic channel, the second pneumatic channel applying pneumatic pressure to the fourth portion of the third fluid channel to thereby deform the fourth portion against the first layer and thereby communicate fluid through the third fluid channel.