NL2036031B1

NL2036031B1 - Apparatus and method for three-dimensional fluid flow focusing

Info

Publication number: NL2036031B1
Application number: NL2036031A
Authority: NL
Inventors: Chou Hou-Pu; Wang Cheng; Zhang Jie
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 channel to receive a ﬂuid containing particles, a second ﬂuidic channel to receive a ﬂow control ﬂuid, and a third ﬂuidic channel to receive the ﬂow control ﬂuid. The first ﬂuidic channel, the second ﬂuidic channel, and the third ﬂuidic channel join together at a junction. A first outlet channel extends from the junction to convey ﬂuid from the junction. A ﬂow focusing feature is positioned at the junction to cause the ﬂow control ﬂuid from the second ﬂuidic channel and the third ﬂuidic channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the first outlet channel as the particles ﬂow along a second horizontal dimension through the outlet channel. ] O l 5

Description

APPARATUS AND METHOD FOR THREE-DIMENSIONAL FLUID FLOW FOCUSING

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 a perspective view of an example junction of the fluid input region of FIG. 7.

[0010] FIG. 8A depicts a perspective view of an example pair of junctions that may be incorporated into another example fluid input region of the cartridge of FIG. 2.

[0011] FIG. 9 depicts a cross-sectional view of the junction of FIG. 8, taken along 9-9 of FIG. 8.

[0012] FIG. 10 depicts a cross-sectional view of the junction of FIG. 8, taken along 10-10 of FIG. 8.

[0013] FIG. 11 depicts a top schematic view of an example of fluid flow dynamics achieved through the junction of FIG. 8.

[0014] FIG. 12 depicts a bottom schematic view of the fluid flow dynamics of FIG. 11.

[0015] FIG. 13 depicts a side schematic view of the fluid flow dynamics of FIG. 11.

[0016] FIG. 14 depicts an end schematic view of the fluid flow dynamics of FIG. 11.

[6017] FIG. 15 depicts an enlarged view of an example mixing structure of the fluid input region of

FIG. 7.

[0018] FIG. 16 depicts a cross-sectional view of the mixing structure of FIG. 15, taken along line 16- 16 of FIG. 15.

[0019] FIG. 17 depicts a perspective schematic view of a portion of the mixing structure of FIG. 15.

[0020] FIG. 18A depicts a schematic view of an example of fluid flow dynamics in a portion of the fluid input region of FIG. 7 just upstream of the mixing structure of FIG. 15.

[0021] FIG. 18B depicts a schematic view of an example of fluid flow dynamics in a portion of the fluid input region of FIG. 7 just downstream of the mixing structure of FIG. 15.

[0022] FIG. 19 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

[0023] 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.

[0024] L Terminology

[0025] 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.9g., 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.

[0026] 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 abbreviated as */”.

[0027] 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.

[0028] 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.

[0029] 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.

[0030] 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.

[0031] 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 119). 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. 5 [0932] 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 perfectly 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.

[0033] The term “perpendicular” shall be understood to include arrangements where one element (eg, 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.).

[0034] 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. Tt 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.

[0035] 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.

[0036] 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.

[6037] 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.

[0038] 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 comer(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.

[0039] 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).

[0040] IL Example of Cell Analysis System

[0041] 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.

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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 200x. 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 us to approximately 1 ms. In other examples, camera (142) may provide any other suitable exposure time.

[6046] 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.

[0047] 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 classity 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.

[6048] 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 subseguent 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.

[6049] 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).

[0050] u. Example of Cartridge

[6051] 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).

[0052] 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), fluid 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.

[0053] 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).

[0054] 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 mput 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). Well openings (440), well channels (340), and top surface (502) of layer (500) thus cooperate 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).

[0055] 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).

[0056] 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 fluid 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.

[6957] A fluidic channel (464) extends between each fluidic flush input regions (460) and a corresponding fluidic flush output region (462), In some scenarios, a fluid input conduit may be coupled with the fluid input port (310) over fluidic flush input regions (460), and a fluid output conduit may be coupled the fluid output port (312) over fluidic flush output region (462).

Fluid may be communicated into flush input region (460) via fluid input ports (310, 410) over fluidic flush input region (460), may flow along fluidic channel (464), then may exit fluidic output region (462) via fluid output ports (312, 412) over fluidic 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.

[0058] 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).

[06059] 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 (800) 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 (800); while objective lens assembly (140) and camera (142) are positioned under cartridge (200) to capture images of cells within imaging region (800). Other examples may provide imaging region (800) at any other suitable location or locations along sampling channel (650). Some examples may also provide a plurality of imaging regions (800) at different respective positions along sampling channel (650).

[0060] Sampling channel (650) terminates in another junction (660), which allows fluid to flow from sampling channel to either a first outlet channel (662) or a second outlet channel (664). Each outlet channel (662, 664) may be selectively opened and closed through pneumatic valving. In particular, a pneumatic channel (322) is positioned over each outlet channel (662, 664), in a region just downstream of junction (660), such that pneumatic pressurization within a selected pneumatic channel (332) causes the underlying region of layer (400) to deform downwardly against top surface (502) of layer (500), thereby effectively providing a closed valve in that outlet channel (662, 664). When the pneumatic pressure is relieved in the pneumatic channel (322) over a given outlet channel (662, 664), the underlying region of layer (400) may return to a non-deformed (i.e, flat) state, thereby effectively opening the fluid pathway through that outlet channel (662, 664).

[0061] If the pneumatic valve for outlet channel (662) is opened while the pneumatic valve for outlet channel (664) is closed, the fluid from sampling channel (650) may flow through outlet channel (662) via junction (660). The fluid path from outlet channel (662) then continues through a selected fluidic channel (442). Each fluidic channel (442) provides a fluidic communication path between outlet channel (662) and a respective well formed by well openings (440) and well channels (340). The fluid path between outlet channel (662) and each fluidic channel (442) may be selectively opened and closed through pneumatic valving as described above. In particular, a pneumatic channel (322) is positioned over each fluidic channel (442), in a region just downstream of outlet channel (662), such that pneumatic pressurization within a selected pneumatic channel (332) causes the underlying region of layer (400) to deform downwardly against top surface (502) of layer (500), thereby effectively providing a closed valve in that fluidic channel (442). When the pneumatic pressure is relieved in pneumatic channel (322), the underlying region of layer (400) may return to a non-deformed (i.e, flat) state, thereby effectively opening the fluid pathway through fluidic channel (442).

[0062] In some scenarios, it may be beneficial to provide additional assistance to the flow of fhad from outlet channel (662) through a selected fluidic channel (442) to reach a selected well channel (340) via an underlying well opening (440). To that end, 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). 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. 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 (442); and ultimately to the selected well channel (340) via an underlying well opening (440). 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.

[0063] If the pneumatic valve for outlet channel (664) is opened while the pneumatic valve for outlet channel (662) is closed, the fluid from sampling channel (650) may 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.

[0064] As noted above, a first set of pneumatic valves may be actuated to allow fluid to flow from sample channel (650) to either outlet channel (662) or outlet channel (664); and if the fluid flows through outlet channel (662); a second set of pneumatic valves may be actuated to allow the fluid to flow through a selected fluidic channel (442) to reach a selected well channel (340) via an underlying well opening (440). System (100) may execute a control algorithm to automatically select which pneumatic valves 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 (800), and the pneumatic valves 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 to a first well channel (340), cells of a second type (as identified by image processing module {144)) may be routed via pneumatic valving to a second well channel (340). and cells of a third type (as identified by image processing module (144)) may be routed via pneumatic valving to sample output region (680).

[0065] IV. Example of Flow Control Features of Cartridge

[0066] 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.

[0067] 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. 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).

[0068] 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 (602) 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, fhadic 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

[6069] 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 (800), 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.

[0070] A. Example of Three-Dimensional Flow Focusing Feature in Junction

[0971] Flow focusing is a concept in fluid dynamics where a focusing fluid or sheath fluid is used to control the flow of another fluid, which may be referred to as a core fluid or a focused fluid.

Flow focusing may be achieved along one or more dimensions in space. Flow focusing may be inertial and/or hydrodynamic. In the context of sampling channel (650), flow focusing may be used to position the cells in the fluid received via sample fluid receiving region (600) within a certain desired region of the cross-sectional space of sampling channel (650), which may facilitate imaging of those cells by providing a consistent and predicable focal plane that does not require movement or refocusing of camera (142) and/or objective lens assembly (140), etc.

Thus, the fluid received via sample fluid receiving region (600) may constitute the core fluid or the focused fluid; while the fluid received via flow control fluid receiving region (620) may constitute the focusing fluid or sheath fluid.

[0072] The flow focusing in sampling channel (650) may be achieved through the structural configuration of junction (640). As shown in FIGS. 8-10, junction (640) includes an expanded recess (642), which extends along a portion of the length of each outlet region (638) of each fluidic channel (626). Each outlet region (638) and fluidic channel (604) have the same first width (WI). Each outlet region (638) and fluidic channel (604) also have the same first height (H1) with respect to top surface (502) of layer (500). Expanded recess (642) includes sidewalls (646) that define a second width (W2) and a length (1). Expanded recess (642) defines second height (H2) between a ceiling (644) of expanded recess (642) and top surface (502) of layer (500).

[9073] The length (L) of expanded recess (642) is greater than the width (W2) of expanded recess (642), such that expanded recess (642) has a rectangular configuration in this example. The width (W2) of expanded recess (642) is slightly smaller than the width (W1) of outlet region (638) and fluidic channel (604), though widths (W1, W2) may be equal in some other examples.

In some examples, each width (W 1, W2) is approximately 50 um. The length {L) of expanded recess (642) is substantially greater than the width (WI) of outlet region (638) and fluidic channel (604). In some examples, the length (L) is approximately 300 um. The height (H2) of expanded recess (642) is also substantially greater than the height (H1) of outlet region (638) and fluidic channel (604). In some examples, the height (H2) of expanded recess (642) ranges from approximately 50 um to approximately 60 um; while the height (H1) of outlet region (638) is approximately 22 pm. In some examples, the length (L) of expanded recess (642) ranges from approximately 4 times the width (W2) of expanded recess (642) to approximately 20 times the width (W2) of expanded recess (642). In some examples, the length (L) of expanded recess (642) ranges from approximately 4 times the width (W 1) of outlet region (638) to approximately 20 times the width (W1) of outlet region (638). In some examples, the height (H2) of expanded recess (642) ranges from approximately 2 times the height (H1) of outlet region (638) and fluidic channel (604) to approximately 4 times the height (H1) of outlet region (638) and fluidic channel (604). The specific dimensions and ratios provided above are not intended to be limiting. Other dimensions and ratios may be used.

[0074] FIGS. 11-14 show an example of flow focusing fluid dynamics that may be achieved by expanded recess (642) at junction (640). As can be seen, fhud enters junction (640) from outlet regions (638) of fluidic channels (626) in opposing directions along the y-dimension; while fluid enters junction (640) from fluidic channel (604) along the x-dimension. The opposing flow of fluid from fluidic channels (626) along the y-dimension thus drives the cells in the fluid of fluidic channel (604) inwardly along the y-dimension, toward the central region of junction (640). This aspect of the flow is best seen in FIGS. 12 and 14. The fluid from fluidic channels (626) also flows upwardly along the z-dimension into expanded recess (642), as best seen in

FIG. 11. The flow of fluid from fluidic channels (626) in expanded recess (642) is turbulent, with sidewalls (646) and ceiling (644) causing the fluid from fluidic channels (626) to flow back downwardly along the z-dimension. This downward flow further drives the cells in the fluid of fluidic channel (604) downwardly along the z-dimension, as best seen in FIGS. 13 and 14.

[6075] Thus, as the cell-containing fluid flows along sampling channel (650) downstream of junction

(640), the cells in the cell-containing fluid may be substantially confined within the z- dimension (along the bottom of sampling channel (650)) and within the y-dimension (along the center of sampling channel (650)). Moreover, this flow focusing may be maintained along the length of sampling channel (650), including within imaging region (800), through the inertial focusing phenomenon. This may be achieved in a flow having a Reynolds number that is approximately 2.5, which may be achieved when the flow rate through fluidic channel (604) is approximately 1 uL/min, the flow rate through each outlet region (638) of each fluidic channel (626) is approximately 9 ul/min, and the flow rate through sampling channel (650) is approximately 10 ul/min.

[0076] While expanded recess (642) is provided in the upper region of junction (640) in this example, some variations of cartridge (200) may instead position expanded recess (642) in the lower region of junction (640). Such repositioning of expanded recess (642) may tend to focus the cells in the upper region of sampling channel (650) rather than the lower region of sampling channel (650). i5 [0077] As another example, a cartridge (200) may include two or more expanded recesses (642), from the same side or from different sides of sampling channel (650). An example of such a configuration is shown in FIG. 8A. In this example, a fluidic channel (1604) is structurally configured and operable like fluidic channel (604) described above; while a set of fluidic channels (1638, 1738) are structurally configured and operable like fluidic channels (626) described above. Fluidic channels (1604, 1638) converge at a junction (1640) which is structurally configured and operable like junction (640) described above. Junction (1640) of this example thus includes an expanded recess (1642), which is structurally configured and operable like expanded recess (642) described above. The fluid output from junction (1640) is conveyed along another fluidic channel (1651), which converges with fluidic channels (1738) at another junction (1740). A sampling channel (1650) provides a fluid output from junction (1740). Sampling channel (1650) may be configured and operable like sampling channel (650) described above.

[0078] Junction (1740) of this example includes an expanded recess (1742) that is structurally configured and operable similar to expanded recess (1642), though expanded recess (1742) extends below the plane of fluidic channels (1604, 1638, 1651, 1738) along the z-dimension {whereas expanded recess (1642) extends above the plane of fluidic channels (1604, 1638, 1651, 1738) along the z-dimension). Thus, to the extent that expanded recess (1642) may tend to focus the flow of fluid downwardly along the z-dimension; expanded recess (1742) may tend to focus the flow of fluid upwardly along the z-dimension. In addition, the combination of recesses (1642, 1742) may tend to focus the flow along an x-y plane that is substantially centered along the z-dimension in sampling channel (1650). While recess (1742) is positioned downstream of recess (1642) in this example, other examples may provide recess (1642) downstream of recess (1742).

[0079] B. Examples of Mixing Structures in Side Channels Upstream of Junction

[0080] 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 (800). 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 (800) 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). ln 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).

[0081] 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).

[0082] As noted above, each fluidic channel (626) includes a herringbone region (630). FIGS. 15-17 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. 17, sidewalls (636) define a length (1.2) of approximately 60 pm. Sidewalls (636) may also form angles (9) 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.

[6083] FIG. 18A shows an example of an arrangement of beads (652) in fluidic channel (626) upstream of herringbone region (630); while FIG. 18B 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 above (and achieve an arrangement like that shown in FIG. 14) 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 (800) that would be achieved by cells later passing through imaging region (800). Herringbone region (630) may thus facilitate replication of cell arrangements in imaging region (800) by bead (660) arrangements in imaging region (800) during calibration of system (100).

[0084] 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).

[0085] C. Examples of Flow Resistance Relationships in Channels of Junction

[0086] In some examples, 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 (800), 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 expanded recess (642) at junction (640) may tend to complicate the achievement of a stable flow rate through sampling channel (650).

[0087] In the present example, the stability of flow along sampling channel (650) is provided, 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).

[0088] In the present example, channels (604, 626, 650) are configured such that the flow resistance of fluidic channel (604) is substantially lower than the flow resistance of channels (626, 650); and such that the flow resistance of each fluidic channel (626) is substantially equal to (or on the same order as) the flow resistance of sampling channel (650). The substantially lower flow resistance 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 of each fluidic channel (626) ranges from approximately 10 times the flow resistance of fluidic channel (604) to approximately 50 times the flow resistance of fluidic channel (604); or more particularly, may be approximately 20 times the flow resistance of fluidic channel (604). In some examples, the flow resistance of sampling channel (650) ranges from being approximately equal to the flow resistance of each fluidic channel (626) to being approximately $ times the flow resistance of each fluidic channel (626); or more particularly, may be approximately 3 times the flow resistance of each fluidic channel (626).

[0089] Minimizing the flow resistance of fluidic channel (604) may minimize the fluid pressure 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 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.

[6090] V. Example of Another Cartridge

[0991] FIG. 19 shows an example of another cartridge (1000) that may be used with system (100) in place of cartridge (200). Except as otherwise noted below, cartridge (1000) of this example may be structurally configured and operable like cartridge (200). Cartridge (1000) of this example includes a sample fluid receiving region (1002) and a flow control fluid receiving region (1004). A flow channel (1006) leads from sample fluid receiving region (1002) to a junction (1006). A pair of flow channels (1008) lead from flow control fluid receiving region (1004) to junction (1006). A sampling channel (1012) extends from junction (1006) to a bend (1014), which leads to a sample output region (1018) via an outlet channel (1016).

[6092] A cell containing fluid may be communicated through sample fluid receiving region (1002), and a flow control fluid may be communicated through flow control fluid receiving region (1094). These two fluids meet at junction (1010), which may provide a flow focusing feature like expanded recess (642), with the merged fluids being conveyed along sampling channel (1012). Light source (130) and optical assembly (132) illuminate imaging region {1020); while objective lens assembly (140) and camera (142) capture images of cells within imaging region (1020). The fluid continues around bend (1014), through outlet channel (1016), and exits cartridge (1000) at sample output region (1018). The fluid that exits cartridge (1000) at sample output region (1018) may be further conveyed to a reservoir that is either integrated into the instrument of system (100) or is external to the instrument.

[0093] Cartridge (1000) is thus similar to cartridge (200) except that cartridge (1000) 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 (1000).

[0094] VI. Examples of Combinations

[0095] 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.

[0096] Example 1

[6097] An apparatus, comprising: a first fluidic channel to receive a fluid containing particles; a second fluidic channel to receive a flow control fluid; a third fluidic channel to receive the flow control fluid; a junction, the first fluidic channel, the second fluidic channel, and the third fluidic channel joining together at the junction; a first outlet channel extending from the junction to convey fluid from the junction; and a flow focusing feature positioned at the junction, the flow focusing feature to cause the flow control fluid from the second fluidic channel and the third fluidic channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the first outlet channel as the particles flow along a second horizontal dimension through the outlet channel.

[0098] Example 2

[0099] The apparatus of Example 1, the second fluidic channel and the third fluidic channel having mirror symmetry with each other.

[00100] Example 3

[60101] The apparatus of any of Examples 1 through 2, the flow focusing feature comprising a recess positioned above the junction.

[00102] Example 4

[00103] The apparatus of Example 3, the recess extending over a portion of the second fluidic channel.

[00104] Example 5

[60105] The apparatus of any of Examples 3 through 4, the recess extending over a portion of the third fluidic channel.

[00106] Example 6

[00107] The apparatus of any of Examples 3 through 5, the first fluidic channel having a width extending along the horizontal dimension, the recess having a length extending along the horizontal dimension, the length of the recess being greater than the width of the first fluidic channel.

[00108] Example 7

[00109] The apparatus of any of Examples 3 through 6, the recess having a rectangular shape.

[00110] Example 8

[00111] The apparatus of any of Examples 1 through 7, the first {fluidic channel extending along the second horizontal dimension.

[00112] Example 9

[00113] The apparatus of any of Examples 1 through 8, the first outlet channel extending along the second horizontal dimension.

[00114] Example 10

[00115] The apparatus of any of Examples 1 through 9, the first fluidic channel and the second fluidic channel being aligned with each other along the second horizontal dimension.

[00116] Example 11

[00117] The apparatus of any of Examples 1 through 10, the second fluidic channel extending along the first horizontal dimension.

[00118] Example 12

[00119] The apparatus of any of Examples 1 through 11, the third fluidic channel extending along the first horizontal dimension.

[00120] Example 13

[00121] The apparatus of any of Examples 1 through 12, the second fluidic channel and the third fluidic channel being aligned with each other along the first horizontal dimension.

[00122] Example 14

[00123] The apparatus of any of Examples 1 through 13, the first fluidic channel being perpendicular to the second fluidic channel at the junction.

[00124] Example 15

[00125] The apparatus of any of Examples 1 through 14, the first fluidic channel being perpendicular to the third fluidic channel at the junction.

[00126] Example 16

[00127] The apparatus of any of Examples 1 through 15, the first outlet channel being perpendicular to the second fluidic channel at the junction.

[00128] Example 17

[00129] The apparatus of any of Examples 1 through 16, the first outlet channel being perpendicular to the third fluidic channel at the junction.

[00130] Example 18

[00131] The apparatus of any of Examples 1 through 17, the flow focusing feature to cause the flow control fluid from the second fluidic channel and the third fluidic channel to flow in a first direction along the vertical dimension and then in a second direction along the vertical dimension to thereby focus the position of the particles in the second direction along the vertical dimension within the first outlet channel as the particles flow along a second horizontal dimension.

[00132] Example 19

[60133] The apparatus of any of Examples 1 through 18, the flow control fluid from the second fluidic channel to focus the position of the particles in a first direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension.

[90134] Example 20

[60135] The apparatus of any of Examples 1 through 19, the flow control fluid from the third fluidic channel to focus the position of the particles in a second direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension.

[00136] Example 21

[00137] The apparatus of any of Examples 1 through 20, the first fluidic channel having a first fluid flow resistance, the second fluidic channel having a second fluid flow resistance, the third fluidic channel having a third fluid flow resistance, the first outlet channel having a fourth fluid flow resistance; and the first fluid flow resistance being lower than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance.

[00138] Example 22

[00139] The apparatus of Example 21, the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance being approximately equal to each other.

[00140] Example 23

[00141] The apparatus of any of Examples 1 through 22, the second fluidic channel including a mixing feature, and the third fluidic channel including a mixing feature.

[00142] Example 24

[00143] The apparatus of Example 23, the mixing feature of the second fluidic channel including a set of recesses, and the mixing feature of the third fluidic channel including a set of recesses.

[00144] Example 25

[00145] The apparatus of Example 24, the set of recesses of the second fluidic channel being disposed over an upper surface of the second fluidic channel, and the set of recesses of the third fluidic channel being disposed over an upper surface of the third fluidic channel.

[00146] Example 26

[00147] The apparatus of any of Examples 24 through 25, the set of recesses of the second fluidic channel including chevron shaped recesses, and the set of recesses of the third fluidic channel including chevron shaped recesses.

[90148] Example 27

[00149] The apparatus of any of Examples 23 through 26, the mixing feature of the second fluidic channel to provide randomized arrangement of particles flowing through the second fluidic channel, and the mixing feature of the third fluidic channel to provide randomized arrangement of particles flowing through the third fluidic channel.

[00150] Example 28

[00151] The apparatus of any of Examples 1 through 27, further comprising a plurality of layers, each of the first fluidic channel, the second fluidic channel, the third fluidic channel, the junction, the first outlet channel, and the flow focusing feature extending through or along one or more of the layers of the plurality of layers.

[00152] Example 29

[00153] The apparatus of Example 28, at least one layer of the plurality of layers comprising polydimethylsiloxane (PDMS).

[00154] Example 30

[60155] The apparatus of Example 29, a first layer of the plurality of layers comprising PDMS having a first thickness, and a second layer of the plurality of layers comprising PDMS having a second thickness.

[00156] Example 31

[00157] The apparatus of any of Examples 28 through 30, the plurality of layers comprising a layer of glass.

[00158] Example 32

[00159] The apparatus of any of Examples 28 through 31, each of the first fluidic channel, the second fluidic channel, the third fluidic channel, the junction, the first outlet channel, and the flow focusing feature comprising one or more respective recesses extending along one layer of the plurality of layers.

[00160] Example 33

[00161] The apparatus of any of Examples 1 through 32, further comprising an imaging region along the first outlet channel to image particles conveyed along the first outlet channel.

[00162] Example 34

[00163] The apparatus of any of Examples 1 through 33, further comprising: a second outlet channel; a third outlet channel; and a sorting junction at an end of the first outlet channel, the sorting junction leading to the second outlet channel and the third outlet channel.

[00164] Example 35

[00165] The apparatus of Example 34, further comprising a first valve and a second valve, the first valve to selectively permit or prevent flow of fluid through the second outlet channel, the second valve to selectively permit or prevent flow of fluid through the third outlet channel.

[00166] Example 36

[00167] The apparatus of any of Examples 34 through 35, further comprising: a plurality of wells; and a plurality of additional fluidic channels, each additional fluidic channel of the plurality of additional fluidic channels leading to a respective well of the plurality of wells.

[00168] Example 37

[00169] The apparatus of Example 36, further comprising a plurality of valves, each valve of the plurality of valves to selectively permit or prevent flow of fluid through a respective additional fluidic channel of the plurality of additional fluidic channels.

[00170] Example 38

[00171] The apparatus of any of Examples 1 through 37, further comprising a sample fluid receiving region, the first fluidic channel to receive a fluid containing particles from an external source via the sample fluid receiving region.

[00172] Example 39

[60173] The apparatus of Example 38, the sample fluid receiving region including a recess having a teardrop shape.

[00174] Example 40

[00175] The apparatus of any of Examples 38 through 39, the sample fluid receiving region including a plurality of filtering elements.

[00176] Example 41

[60177] A method comprising: flowing a fluid containing particles along a first fluidic channel; flowing a flow control fluid along a second fluidic channel while simultaneously flowing the fluid containing particles along the first fluidic channel; flowing the flow control fluid along a third fluidic channel while simultaneously flowing the fluid containing particles along the first fluidic channel and while simultaneously flowing the flow control fluid along the second fluidic channel; and allowing the flow of the fluid containing particles along the first fluidic channel to join the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel at a junction, thereby forming a joined fluid, the junction including an outlet channel and a flow focusing feature; the flow focusing feature causing the flow control fluid from the second fluidic channel and the third fluidic channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the outlet channel as {he particles flow in the joined fluid along a second horizontal dimension through the outlet channel.

[00178] Example 42

[60179] The method of Example 41, the flow focusing feature comprising a recess positioned above the junction, allowing the flow of the fluid containing particles along the first fluidic channel to join the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel at the junction comprising allowing the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel to enter the recess positioned above the junction.

[00180] Example 43

[00181] The method of Example 42, the flow control fluid from the second fluidic channel and the flow control fluid from the third fluidic channel flowing upwardly into the recess then downwardly toward the outlet channel.

[00182] Example 44

[00183] The method of any of Examples 41 through 43, flowing the fluid containing particles along the first fluidic channel including flowing the fluid containing particles along the second horizontal dimension.

[00184] Example 45

[00185] The method of any of Examples 41 through 44, flowing the flow control fluid along the second fluidic channel including flowing the flow control fluid along the first horizontal dimension into the junction.

[00186] Example 46

[00187] The method of Example 45, flowing the flow control fluid along the second fluidic channel further including flowing the flow control fluid along a serpentine path before flowing the flow control fluid along the first horizontal dimension into the junction.

[00188] Example 47

[00189] The method of any of Examples 41 through 46, flowing the flow control fluid along the third fluidic channel including flowing the flow control fluid along the first horizontal dimension into the junction.

[00190] Example 48

[00191] The method of Example 47, flowing the flow control fluid along the third fluidic channel further including flowing the flow control fluid along a serpentine path before flowing the flow control fluid along the first horizontal dimension into the junction.

[00192] Example 49

[00193] The method of any of Examples 41 through 48, the flow control fluid from the second fluidic channel focusing the position of the particles in a first direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

[00194] Example 50

[00195] The method of any of Examples 41 through 49, the flow control fluid from the third fluidic channel focusing the position of the particles in a second direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

[00196] Example 51

[60197] The method of any of Examples 41 through 50, the fluid containing particles flowing along the first fluidic channel at a first fluid flow resistance; the flow control fluid flowing along the second fluidic channel at a second fluid flow resistance; the flow control fluid flowing along the third fluidic channel at a third fluid flow resistance; the joined fluid flowing through the outlet channel at a fourth fluid flow resistance; and the first fluid flow resistance being lower than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance.

[00198] Example 52

[00199] The method of Example 51, the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance being approximately equal to each other.

[00200] Example 53

[00201] The method of any of Examples 41 through 52, flowing the flow control fluid along the second fluidic channel including flowing the flow control fluid through a mixing feature of the second fluidic channel; and flowing the flow control fluid along the third fluidic channel including flowing the flow control fluid through a mixing feature of the third fluidic channel.

[00202] Example 54

[00203] The method of Example 53, the mixing feature of the second fluidic channel including a set of recesses, flowing the flow control fluid through the mixing feature of the second fluidic channel including flowing the flow control fluid along the set of recesses of the mixing feature of the second fluidic channel; and the mixing feature of the third fluidic channel including a set of recesses, flowing the flow control fluid through the mixing feature of the third fluidic channel including flowing the flow control fluid along the set of recesses of the mixing feature of the third fluidic channel.

[00204] Example 55

[00205] The method of Example 54, the set of recesses of the second fluidic channel being disposed over an upper surface of the second fluidic channel, flowing the flow control fluid along the set of recesses of the mixing feature of the second fluidic channel including flowing the flow control fluid upwardly into the set of recesses of the mixing feature of the second fluidic channel; and the set of recesses of the third fluidic channel being disposed over an upper surface of the third fluidic channel, flowing the flow control fluid along the set of recesses of the mixing feature of the third fluidic channel including flowing the flow control fluid upwardly into the set of recesses of the mixing feature of the third fluidic channel.

[00206] Example 56

[60207] The method of any of Examples 53 through 55, the mixing feature of the second fluidic channel providing randomized arrangement of particles flowing through the second fluidic channel, the mixing feature of the third fluidic channel providing randomized arrangement of particles flowing through the third fluidic channel.

[00208] Example 57

[00209] The method of any of Examples 41 through 56, further comprising capturing images of particles as the particles are conveyed along the outlet channel.

[00210] Example 58

[00211] The method of Example 57, further comprising classifying the particles by analyzing the captured images.

[00212] Example 59

[00213] The method of Example 58, further comprising sorting particles from the outlet channel in response to classifying or identifying the particles by analyzing the captured images.

[00214] Example 60

[00215] The method of Example 59, sorting particles including activating one or more valves to redirect fluid flow from the outlet channel to another outlet channel selected from a plurality of other outlet channels.

[00216] Example 61

[00217] The method of any of Examples 41 through 60, the particles including biological cells.

[00218] VII. Miscellaneous

[00219] 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.

[00220] 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.

[00221] 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.

[00222] 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.

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

Dutch-language claims:

CLAUSES i. An apparatus, comprising: a first fluidic channel to receive a fluid containing particles; a second fluidic channel to receive a flow control fluid; a third fluidic channel to receive the flow control fluid; a junction, the first fluidic channel, the second fluidic channel, and the third fluidic channel joining together at the junction; a first outlet channel extending from the junction to convey fluid from the junction; and a flow focusing feature positioned at the junction, the flow focusing feature to cause the flow control fluid from the second fluidic channel and the third fluidic channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the first outlet channel as the particles flow along a second horizontal dimension through the outlet channel. 2. The apparatus of clause 1, the second fluidic channel and the third fluidic channel having mirror symmetry with each other.

3. The apparatus of any of clauses 1 through 2, the flow focusing feature comprising a recess positioned above the junction, 4. The apparatus of clause 3, the recess extending over a portion of the second fluidic channel. 5. The apparatus of any of clauses 3 through 4, the recess extending over a portion of the third fluidic channel.

6. The apparatus of any of clauses 3 through 5, the first fluidic channel having a width extending along the horizontal dimension, the recess having a length extending along the horizontal dimension, the length of the recess being greater than the width of the first fluidic channel. 7. The apparatus of any of clauses 3 through 6, the recess having a rectangular shape. 8. The apparatus of any of clauses 1 through 7, the first fluidic channel extending along the second horizontal dimension. 9. The apparatus of any of clauses 1 through 8, the first outlet channel extending along the second horizontal dimension. 10. The apparatus of any of clauses | through 9, the first fluidic channel and the second fluidic channel being aligned with each other along the second horizontal dimension.

11. The apparatus of any of clauses | through 10, the second fluidic channel extending along the first horizontal dimension. 12. The apparatus of any of clauses 1 through 11, the third fluidic channel extending along the first horizontal dimension. 13. The apparatus of any of clauses 1 through 12, the second fluidic channel and the third fluidic channel being aligned with each other along the first horizontal dimension.

14. The apparatus of any of clauses 1 through 13, the first fluidic channel being perpendicular to the second fluidic channel at the junction. 15. The apparatus of any of clauses 1 through 14, the first fluidic channel being

S perpendicular to the third fluidic channel at the junction. 16. The apparatus of any of clauses 1 through 15, the first outlet channel being perpendicular to the second fluidic channel at the junction. 17. The apparatus of any of clauses 1 through 16, the first outlet channel being perpendicular to the third fluidic channel at the junction. 18. The apparatus of any of clauses 1 through 17, the flow focusing feature to cause the flow control fluid from the second fluidic channel and the third fluidic channel to flow in a first direction along the vertical dimension and then in a second direction along the vertical dimension to thereby focus the position of the particles in the second direction along the vertical dimension within the first outlet channel as the particles flow along a second horizontal dimension. 19. The apparatus of any of clauses 1 through 18, the flow control fluid from the second fluidic channel to focus the position of the particles in a first direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension. 20. The apparatus of any of clauses 1 through 19, the flow control fluid from the third fluidic channel to focus the position of the particles in a second direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension. 21. The apparatus of any of clauses 1 through 20, the first fluidic channel having a first fluid flow resistance, the second fluidic channel having a second fluid flow resistance, the third fluidic channel having a third fluid flow resistance, the first outlet channel having a fourth fluid flow resistance; and the first fluid flow resistance being lower than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance.

22. The apparatus of clause 21, the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance being approximately equal to each other. 23. The apparatus of any of clauses 1 through 22, the second fluidic channel including a

S mixing feature, and the third fluidic channel including a mixing feature. 24. The apparatus of clause 23, the mixing feature of the second fluidic channel including a set of recesses, and the mixing feature of the third fluidic channel including a set of recesses. 25. The apparatus of clause 24, the set of recesses of the second fluidic channel being disposed over an upper surface of the second fluidic channel, and the set of recesses of the third fluidic channel being disposed over an upper surface of the third fluidic channel. 26. The apparatus of any of clauses 24 through 25, the set of recesses of the second fluidic channel including chevron shaped recesses, and the set of recesses of the third fluidic channel including chevron shaped recesses. 27. The apparatus of any of clauses 23 through 26, the mixing feature of the second fluidic channel to provide randomized arrangement of particles flowing through the second fluidic channel, and the mixing feature of the third fluidic channel to provide randomized arrangement of particles flowing through the third fluidic channel. 28. The apparatus of any of clauses 1 through 27, further comprising a plurality of layers, each of the first fluidic channel, the second fluidic channel, the third fluidic channel, the junction, the first outlet channel, and the flow focusing feature extending through or along one or more of the layers of the plurality of layers. 29. The apparatus of clause 28, at least one layer of the plurality of layers comprising polydimethylsiloxane (PDMS). 30. The apparatus of clause 29, a first layer of the plurality of layers comprising PDMS having a first thickness, and a second layer of the plurality of layers comprising PDMS having a second thickness.

31. The apparatus of any of clauses 28 through 30, the plurality of layers comprising a layer of glass. 32. The apparatus of any of clauses 28 through 31, each of the first fluidic channel, the second fluidic channel, the third fluidic channel, the junction, the first outlet channel, and the flow focusing feature comprising one or more respective recesses extending along one layer of the plurality of layers. 33. The apparatus of any of clauses 1 through 32, further comprising an imaging region along the first outlet channel to image particles conveyed along the first outlet channel. 34. The apparatus of any of clauses 1 through 33, further comprising: a second outlet channel; a third outlet channel; and a sorting junction at an end of the first outlet channel, the sorting junction leading to the second outlet channel and the third outlet channel. 35. The apparatus of clause 34, further comprising a first valve and a second valve, the first valve to selectively permit or prevent flow of fluid through the second outlet channel, the second valve to selectively permit or prevent flow of fluid through the third outlet channel. 36. The apparatus of any of clauses 34 through 35, further comprising: a plurality of wells; and a plurality of additional fluidic channels, each additional fluidic channel of the plurality of additional fluidic channels leading to a respective well of the plurality of wells. 37. The apparatus of clause 36, further comprising a plurality of valves, each valve of the plurality of valves to selectively permit or prevent flow of fluid through a respective additional fluidic channel of the plurality of additional fluidic channels. 38. The apparatus of any of clauses | through 37, further comprising a sample fluid receiving region, the first fluidic channel to receive a fluid containing particles from an external source via the sample fluid receiving region.

39. The apparatus of clause 38, the sample fluid receiving region including a recess having a teardrop shape.

40. The apparatus of any of clauses 38 through 39, the sample fluid receiving region including a plurality of filtering elements.

41. A method comprising:

flowing a fluid containing particles along a first fluidic channel; flowing a flow control fluid along a second fluidic channel while simultaneously flowing the fluid containing particles along the first fluidic channel; flowing the flow control fluid along a third fluidic channel while simultaneously flowing the fluid containing particles along the first fluidic channel and while simultaneously flowing the flow control fluid along the second fluidic channel; and allowing the flow of the fluid containing particles along the first fluidic channel to join the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel at a junction, thereby forming a joined fluid, the junction including an outlet channel and a flow focusing feature; the flow focusing feature causing the flow control fluid from the second fluidic channel and the third fluidic channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the outlet channel as the particles flow in the joined fluid along a second horizontal dimension through the outlet channel.

42. The method of clause 41, the flow focusing feature comprising a recess positioned above the junction, allowing the flow of the fluid containing particles along the first fluidic channel to join the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel at the junction comprising allowing the flow of the flow control fluid along the second fluidic channel and the flow of the flow control fluid along the third fluidic channel to enter the recess positioned above the junction.

43. The method of clause 42, the flow control fluid from the second fluidic channel and the flow control fluid from the third fluidic channel flowing upwardly into the recess then downwardly toward the outlet channel.

44. The method of any of clauses 41 through 43, flowing the fluid containing particles along the first fluidic channel including flowing the fluid containing particles along the second horizontal dimension.

45. The method of any of clauses 41 through 44, flowing the flow control fluid along the second fluidic channel including flowing the flow control fluid along the first horizontal dimension into the junction,

46. The method of clause 45, flowing the flow control fluid along the second fluidic channel further including flowing the flow control fluid along a serpentine path before flowing the flow control fluid along the first horizontal dimension into the junction.

47. The method of any of clauses 41 through 46, flowing the flow control fluid along the third fluidic channel including flowing the flow control fluid along the first horizontal dimension into the junction.

48. The method of clause 47, flowing the flow control fluid along the third fluidic channel further including flowing the flow control fluid along a serpentine path before flowing the flow control fluid along the first horizontal dimension into the junction.

49. The method of any of clauses 41 through 48, the flow control fluid from the second fluidic channel focusing the position of the particles in a first direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

50. The method of any of clauses 41 through 49, the flow control fluid from the third fluidic channel focusing the position of the particles in a second direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

51. The method of any of clauses 41 through 50, the fluid containing particles flowing along the first fluidic channel at a first fluid flow resistance;

the flow control fluid flowing along the second fluidic channel at a second fluid flow resistance; the flow control fluid flowing along the third fluidic channel at a third fluid flow resistance; the joined fluid flowing through the outlet channel at a fourth fluid flow resistance; and the first fluid flow resistance being lower than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance. 52. The method of clause 51, the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance being approximately equal to each other.

53. The method of any of clauses 41 through 52, flowing the flow control fluid along the second fluidic channel including flowing the flow control fluid through a mixing feature of the second fluidic channel; and flowing the flow control fluid along the third fluidic channel including flowing the flow control fluid through a mixing feature of the third fluidic channel.

54. The method of clause 53, the mixing feature of the second fluidic channel including a set of recesses, flowing the flow control fluid through the mixing feature of the second fluidic channel including flowing the flow control fluid along the set of recesses of the mixing feature of the second fluidic channel; and the mixing feature of the third fluidic channel including a set of recesses, flowing the flow control fluid through the mixing feature of the third fluidic channel including flowing the flow control fluid along the set of recesses of the mixing feature of the third fluidic channel.

55. The method of clause 54, the set of recesses of the second fluidic channel being disposed over an upper surface of the second fluidic channel, flowing the flow control fluid along the set of recesses of the mixing feature of the second fluidic channel including flowing the flow control fluid upwardly into the set of recesses of the mixing feature of the second fluidic channel; and the set of recesses of the third fluidic channel being disposed over an upper surface of the third fluidic channel, flowing the flow control fluid along the set of recesses of the mixing feature of the third fluidic channel including flowing the flow control fluid upwardly into the set of recesses of the mixing feature of the third fluidic channel.

56. The method of any of clauses 53 through 55, the mixing feature of the second fluidic channel providing randomized arrangement of particles flowing through the second fluidic channel, the mixing feature of the third fluidic channel providing randomized arrangement of particles flowing through the third fluidic channel. 57. The method of any of clauses 41 through 56, further comprising capturing images of

S particles as the particles are conveyed along the outlet channel. 58. The method of clause 57, further comprising classifying the particles by analyzing the captured images. 59. The method of clause 58, further comprising sorting particles [rom the outlet channel in response to classifying or identifying the particles by analyzing the captured images. 60. The method of clause 59, sorting particles including activating one or more valves to redirect fluid flow from the outlet channel to another outlet channel selected from a plurality of other outlet channels. 61. The method of any of clauses 41 through 60, the particles including biological cells.

Claims

CONCLUSIONS

1. An apparatus comprising: a first fluid conduit for receiving fluid containing particles; a second fluid conduit for receiving a flow control fluid; a third fluid conduit for receiving the flow control fluid; a connection, the first fluid conduit, the second fluid conduit and the third fluid conduit converging at the connection; a first outlet conduit extending from the connection for conveying fluid from the connection; and a flow focusing means positioned at the connection, the flow focusing means causing the flow control fluid from the second fluid conduit and the third fluid conduit to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the first outlet conduit as the particles flow along a second horizontal dimension through the outlet conduit.

2. The apparatus of claim 1, wherein the second fluid channel and the third fluid channel have mirror symmetry with each other.

3. Apparatus according to any one of claims | to 2, wherein the current focusing means comprises a recess positioned above the connection.

4. The apparatus of claim 3, wherein the recess extends over a portion of the second fluid channel.

5. Apparatus according to any one of claims 3 to 4, wherein the recess extends over a portion of the third fluid channel.

6. The apparatus of any one of claims 3 to 5, wherein the first fluid channel has a width extending along the horizontal dimension, the recess has a length extending along the horizontal dimension, the length of the recess being greater than the width of the first fluid channel.

7. Device according to any one of claims 3 to 6, wherein the recess has a rectangular shape.

8. The apparatus of any one of claims 1 to 7, wherein the first fluid channel extends S along the second horizontal dimension.

9. The apparatus of any one of claims 1 to 8, wherein the first outlet channel extends along the second horizontal dimension.

10. The apparatus of any one of claims 1 to 9, wherein the first fluid channel and the second fluid channel are aligned with each other along the second horizontal dimension.

11. The apparatus of any one of claims 1 to 10, wherein the second fluid channel extends along the first horizontal dimension.

12. The apparatus of any one of claims 1 to 11, wherein the third fluid channel extends along the first horizontal dimension.

13. The apparatus of any one of claims | to 12, wherein the second fluid channel and the third fluid channel are aligned with each other along the first horizontal dimension.

14. The apparatus of any one of claims 1 to 13, wherein the first fluid channel is perpendicular to the second fluid channel at the connection.

15. The apparatus of any one of claims 1 to 14, wherein the first fluid channel is perpendicular to the third fluid channel at the connection.

16. Apparatus according to any one of claims | to 15, wherein the first outlet channel is perpendicular to the second fluid channel at the connection.

17. The apparatus of any one of claims 1 to 16, wherein the first outlet channel is perpendicular to the third fluid channel at the connection.

18. The apparatus of any one of claims 1 to 17, wherein the flow focusing means causes the flow control fluid from the second fluid channel and the third fluid channel to flow in a first direction along the vertical dimension and then in a second direction along the vertical dimension to thereby focus the position of the particles in the second direction along the vertical dimension within the first outlet channel as the particles flow along a second horizontal dimension.

19. The apparatus of any one of claims 1 to 18, wherein the flow control fluid from the second fluid channel focuses the position of the particles in a first direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension.

20. The apparatus of any one of claims 1 to 19, wherein the flow control fluid from the third fluid channel focuses the position of the particles in a second direction along the first horizontal dimension within the first outlet channel as the particles flow along the second horizontal dimension.

21. The apparatus of any one of claims 1 to 20, wherein the first fluid channel has a first fluid flow resistance, the second fluid channel has a second fluid flow resistance, the third fluid channel has a third fluid flow resistance, and the first outlet channel has a fourth fluid flow resistance; and wherein the first fluid flow resistance is lower than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance.

22. The apparatus of claim 21, wherein the second fluid flow resistance, the third fluid flow resistance and the fourth fluid flow resistance are approximately equal to each other.

23. The apparatus of any one of claims 1 to 22, wherein the second fluid channel comprises a mixing means, and the third fluid channel comprises a mixing means.

24. The apparatus of claim 23, wherein the mixing means of the second fluid channel comprises a set of recesses, and the mixing means of the third fluid channel comprises a set of recesses.

25. The apparatus of claim 24, wherein the set of recesses of the second fluid channel are disposed over an upper surface of the second fluid channel, and the set of recesses of the third fluid channel are disposed over an upper surface of the third fluid channel.

26. The apparatus of any one of claims 24 to 25, wherein the set of recesses of the second fluid channel comprises chevron-shaped recesses, and the set of recesses of the third fluid channel comprises chevron-shaped recesses.

27. The apparatus of any one of claims 23 to 26, wherein the mixing means of the second fluid channel provides for a random arrangement of particles flowing through the second fluid channel, and the mixing means of the third fluid channel provides for a random arrangement of particles flowing through the third fluid channel.

28. The apparatus of any of claims 1 to 27 further comprising a plurality of layers, wherein each of the first fluid channel, the second fluid channel, the third fluid channel, the connection, the first outlet channel and the flow focusing means extends through or along one or more of the layers of the plurality of layers.

29. The device of claim 28, wherein at least one layer of the plurality of layers comprises polydimethylsiloxane (PDMS).

The device of claim 29, wherein a first layer of the plurality of layers comprises PDMS having a first thickness, and a second layer of the plurality of layers comprises PDMS having a second thickness.

31. The apparatus of any one of claims 28 to 30, wherein the plurality of layers comprises a layer of glass.

32. The apparatus of any of claims 28 to 31, wherein each of the first fluid channel, the second fluid channel, the third fluid channel, the connection, the first outlet channel and the flow focusing means comprises one or more respective recesses extending along one of the plurality of layers.

33. The apparatus of any one of claims 1 to 32, further comprising an imaging region along the first outlet channel for imaging particles transported along the first outlet channel.

34. The apparatus of any one of claims 1 to 33, further comprising: a second outlet channel; a third outlet channel; and a sorting connection at an end of the first outlet channel, the sorting connection leading to the second outlet channel and the third outlet channel.

35. The apparatus of claim 34 further comprising a first valve and a second valve, the first valve selectively allowing or preventing fluid flow through the second outlet channel, and the second valve selectively allowing or preventing fluid flow through the third outlet channel.

36. The apparatus of any one of claims 34 to 35 further comprising: a plurality of wells; and a plurality of additional fluid channels, each additional fluid channel of the plurality of additional fluid channels leading to a respective well of the plurality of wells.

37. The apparatus of claim 36 further comprising a plurality of valves, each valve of the plurality of valves selectively permitting or preventing fluid flow through a respective one of the plurality of additional fluid channels.

38. The apparatus of any of claims 1 to 37, further comprising a sample fluid receiving region, wherein the first fluid channel receives fluid containing particles from an external source via the sample fluid receiving region.

39. The apparatus of claim 38, wherein the sample fluid receiving region comprises a recess having a teardrop shape.

40. The apparatus of any one of claims 38 to 39, wherein the sample fluid receiving region comprises a plurality of filter elements.

41. A method comprising:

flowing a fluid containing particles along a first fluid channel; flowing a flow control fluid along a second fluid channel while simultaneously flowing the fluid containing particles along the first fluid channel; flowing the flow control fluid along a third fluid channel while simultaneously flowing the fluid containing particles along the first fluid channel and while simultaneously flowing the flow control fluid along the second fluid channel; and allowing the flow of the fluid containing particles along the first fluid channel to join the flow of the flow control fluid along the second fluid channel and the flow of the flow control fluid along the third fluid channel at an intersection, thereby forming a conjoined fluid, the connection comprising an outlet channel and a flow focusing means; wherein the flow focusing means causes the flow control fluid from the second fluid channel and the third fluid channel to focus the position of the particles along a vertical dimension and along a first horizontal dimension within the outlet channel while the particles in the conjoined fluid flow along a second horizontal dimension through the outlet channel.

42. The method of claim 41, wherein the flow focusing means comprises a recess positioned above the connection, whereby the flow of the fluid containing particles along the first fluid channel may join the flow of the flow control fluid along the second fluid channel and the flow of the flow control fluid along the third fluid channel at the connection, including allowing the flow of the flow control fluid along the second fluid channel and the flow of the flow control fluid along the third fluid channel to enter the recess positioned above the connection.

43. The method of claim 42, wherein the flow control fluid from the second fluid channel and the flow control fluid from the third fluid channel flows upward into the recess and then downward toward the outlet channel.

44. A method according to any one of claims 41 to 43, wherein the fluid containing particles flows along the first fluid channel, including flowing the fluid containing particles along the second horizontal dimension,

45. The method of any one of claims 41 to 44, wherein the flow control fluid flows along the second fluid channel, including flowing the flow control fluid along the first horizontal dimension in the connection.

46. The method of claim 45, wherein the flow control fluid flows along the second fluid channel, further comprising flowing the flow control fluid along a tortuous path before flowing the flow control fluid along the first horizontal dimension in the connection.

47. A method as claimed in any one of claims 41 to 46, wherein the flow control fluid flows along the third fluid channel, including flowing the flow control fluid along the first horizontal dimension in the connection.

48. The method of claim 47, wherein the flow control fluid flows along the third fluid channel, further comprising flowing the flow control fluid along a tortuous path prior to flowing the flow control fluid along the first horizontal dimension in the connection.

49. A method as claimed in any one of claims 41 to 48, wherein the flow control fluid from the second fluid channel focuses the position of the particles in a first direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

50. A method as claimed in any one of claims 41 to 49, wherein the flow control fluid from the third fluid channel focuses the position of the particles in a second direction along the first horizontal dimension within the outlet channel as the particles flow along the second horizontal dimension.

51. A method according to any one of claims 41 to 50, wherein the fluid contains particles flowing along the first fluid channel with a first fluid flow resistance; wherein the flow control fluid flows along the second fluid channel with a second fluid flow resistance; wherein the flow control fluid flows along the third fluid channel with a third fluid flow resistance;

wherein the combined fluid flows through the outlet channel with a fourth fluid flow resistance: and wherein the first fluid flow resistance is less than each of the second fluid flow resistance, the third fluid flow resistance, and the fourth fluid flow resistance.

52. The method of claim 51, wherein the second fluid flow resistance, the third fluid flow resistance and the fourth fluid flow resistance are approximately equal to each other.

53. The method of any one of claims 41 to 52, wherein the flow control fluid flows along the second fluid channel, including flowing the flow control fluid through a mixing means of the second fluid channel; and flowing the flow control fluid along the third fluid channel, including flowing the flow control fluid through a mixing means of the third fluid channel.

54. The method of claim 53, wherein the second fluid channel mixing means comprises a set of recesses, wherein the flow control fluid flows through the second fluid channel mixing means, including flowing the flow control fluid past the set of recesses of the second fluid channel mixing means; and wherein the third fluid channel mixing means comprises a set of recesses, wherein the flow control fluid flows through the third fluid channel mixing means, including flowing the flow control fluid past the set of recesses of the third fluid channel mixing means.

55. The method of claim 54, wherein the set of recesses of the second fluid channel are disposed over a top surface of the second fluid channel, whereby the flow control fluid flows past the set of recesses of the mixing element of the second fluid channel, including flowing the flow control fluid upward into the array of recesses of the mixing means of the second fluid channel; and wherein the array of recesses of the third fluid channel are disposed over a top surface of the third fluid channel, whereby the flow control fluid flows past the array of recesses of the mixing element of the third fluid channel, including flowing the flow control fluid upward into the array of recesses of the mixing means of the third fluid channel.

56. A method according to any one of claims 53 to 55, wherein the mixing means of the second fluid channel provides a random arrangement of particles flowing through the second fluid channel, while the mixing means of the third fluid channel provides a random arrangement of particles flowing through the third fluid channel.

57. A method according to any one of claims 41 to 56, further comprising capturing images of particles as the particles are transported along the outlet channel.

58. The method of claim 57, further comprising classifying the particles by analyzing the captured images.

59. The method of claim 58 further comprising sorting particles from the exhaust duct in response to classifying or identifying the particles by analyzing the captured images.

60. The method of claim 59, wherein sorting particles comprises activating one or more valves to divert fluid flow from the outlet channel to another outlet channel selected from a plurality of other fluid channels.

61. A method according to any one of claims 41 to 60, wherein the particles comprise biological cells.