Methods, systems, and devices for linear electrowetting cartridges

Abstract

A method for preparing droplets on a surface of a cartridge for analysis is disclosed, the droplets including a plurality of particles. The method includes transporting the droplets along a single path on the surface of the cartridge via a plurality of electrodes of the cartridge, the single path being substantially linear, and the plurality of electrodes configured to transport the droplets along the single path on the surface of the cartridge. The method also includes analyzing the droplets at one or more locations along the single path on the surface of the cartridge.

Description

[Technical Field]

[0001] FIELD OF THE DISCLOSURE The present disclosure involves systems and methods for analyzing droplets, including multiple particles, along a defined path on a surface of a cartridge. That is, the disclosed devices and methods transport droplets along a defined path (e.g., a single, generally linear path) on a surface of a cartridge and analyze the droplets to identify parameters (e.g., unique identifying features) of the droplets and / or their components (e.g., at least one paramagnetic barcoded bead). [Background technology]

[0002] (background) Assays (including immunological assays) and other analytical evaluations (e.g., polymerase chain reaction (PCR) tests) can be performed on one or more portions of a sample using a variety of different methods, including by utilizing multiple particles (e.g., paramagnetic barcoded beads) and other components in droplets of solution containing the sample to aid in performing the assays and other analytical evaluations. Summary of the Invention [Means for solving the problem]

[0003] (summary) In some examples, a plurality of particles (e.g., paramagnetic barcoded beads) can be suspended in a solution that can be used for testing and identification of components in the solution and / or portions thereof (e.g., droplets of the solution). To increase the accuracy and speed of test results, it is desirable to ensure that the plurality of particles are properly dispersed throughout the solution prior to testing and are properly mixed and analyzed during the testing protocol (e.g., assay).

[0004] When an operator manually prepares a solution for testing, the homogeneity and number of particles throughout the prepared solution may be inconsistent and / or inaccurate. Furthermore, if an operator allows an excessive amount of time to pass between manually preparing (e.g., stirring) the solution and taking a sample therefrom, the particles may become less homogenized throughout the solution (e.g., by settling to the bottom of the container, clumping together, or both, among other potential issues), which in turn may affect the accuracy and precision of any test results for which the solution may be used. Thus, manual preparation of solutions is subject to variability between preparations and / or operators, thus degrading the accuracy and precision of any associated test results.

[0005] In one embodiment, a method is described for analyzing droplets on a surface of a cartridge, the droplets including a plurality of particles, the method including transporting the droplets along a single path on the surface of the cartridge via a plurality of electrodes of the cartridge, the single path being substantially linear, the plurality of electrodes configured to transport the droplets along the single path on the surface of the cartridge, and analyzing the droplets at one or more locations along the single path on the surface of the cartridge.

[0006] In another example, a non-transitory computer-readable medium having program instructions stored thereon is described, the program instructions, upon execution by a controller, causing the controller to perform a set of operations including transporting a droplet along a single path on a surface of the cartridge via a plurality of electrodes of the cartridge, the droplet including a plurality of particles, the single path being substantially linear, the plurality of electrodes configured to transport the droplet along the single path on the surface of the cartridge, and analyzing the droplet at one or more locations along the single path on the surface of the cartridge.

[0007] The features, functions, and advantages discussed can be achieved independently in various embodiments or may be combined in yet other embodiments. Further details of the embodiments can be found in the following description and with reference to the drawings. [Brief explanation of the drawings]

[0008] The above, as well as additional features, will be better understood through the following illustrative and non-limiting detailed description of exemplary embodiments, with reference to the accompanying drawings.

[0009] [Figure 1] FIG. 1 illustrates a simplified block diagram of an exemplary computing device, in accordance with an exemplary embodiment.

[0010] [Figure 2] FIG. 2 illustrates a cartridge according to an exemplary embodiment.

[0011] [Figure 3] FIG. 3 illustrates a cartridge according to an exemplary embodiment.

[0012] [Figure 4] FIG. 4 illustrates a cartridge according to an exemplary embodiment.

[0013] [Figure 5] FIG. 5 illustrates a method according to an example embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0014] All figures are schematic and not necessarily to scale, and generally show only those parts that are necessary to clarify the exemplary embodiments; other parts may be omitted or only suggested.

[0015] (Detailed explanation) Exemplary embodiments will now be described more fully hereinafter with reference to the accompanying drawings. What is encompassed by the claims may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided as examples. Furthermore, like numbers refer to the same or similar elements or components throughout.

[0016] In embodiments, the present disclosure is directed to devices and methods for manipulating droplets of solution containing a sample and a plurality of particles (e.g., one or more types of paramagnetic barcoded beads) containing one or more identifying features (such as a unique barcode, color, shape, alphanumeric symbol, and / or the like). These particles include one or more of the following: microbeads, microparticles, micropellets, microwafers, microparticles, paramagnetic microparticles, paramagnetic microparticles containing one or more barcodes, and / or beads containing one or more nickel barcodes, all containing one or more identifying features (such as a barcode, color, shape, alphanumeric symbol, and / or the like). The particles may also be magnetic or paramagnetic. Particles suitable for use in the present disclosure are capable of attachment to other substances, such as derivatives, linker molecules, proteins, nucleic acids, or combinations thereof. The ability of the particles to be attached to other substances can result from the particle material and from any additional surface modification or functionalization of the particles. The particles can be functionalized or capable of becoming functionalized for covalent or non-covalent attachment of proteins, nucleic acids, linker molecules, or derivatives as described herein.

[0017] For example, the surfaces of these particles (e.g., paramagnetic barcoded beads) can be modified or functionalized with amines, biotin, streptavidin, avidin, protein A, sulfhydryls, hydroxyls, and carboxyls. These particles can be spherical or other shapes, light-transmitting, and digitally encoded, e.g., with an image that provides high contrast and high signal-to-noise optical detection, to facilitate bead identification. To the extent an image exists, the image can be implemented by a physical structure having a pattern that is partially substantially transparent to light (e.g., transparent, translucent, and / or translucent) and partially substantially opaque to light (e.g., reflective and / or absorbing). The pattern of transmitted light can be determined (e.g., by scanning or imaging), and the code represented by the image on the encoded bead can be decoded. Various code patterns, such as circles, squares, or other geometric shapes, can be designed, as long as they can be recognized by an optical code reader. Examples of one or more of these types of particles can be found in US Pat. Nos. 7,745,091, 8,148,139, and 8,614,852.

[0018] Additionally or alternatively, these particles (e.g., paramagnetic barcoded beads) may comprise one or more materials, including one or more of the following: glass, polymer, polystyrene, latex, elemental metal, ceramic, metal composite, metal alloy, silicon, or other support materials such as agarose, ceramic, glass, quartz, polyacrylamide, polymethyl methacrylate, carboxylic acid-modified latex, melamine, and Sepharose, and / or hybrids of one or more thereof. Particularly useful commercially available materials include carboxylic acid-modified latex, cyanogen bromide-activated Sepharose beads, fused silica particles, isothiocyanate glass, polystyrene, and carboxylic acid monodisperse microspheres. Furthermore, these particles may also comprise one or more specific shapes, dimensions, and / or configurations and be modified for one or more specific uses. For example, the particles (e.g., paramagnetic barcoded beads) may vary in size from about 0.1 microns to about 100 microns, e.g., about 0.1, 0.5, 1.0, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 microns. In a further aspect, the particles may be surface-modified and / or functionalized with biomolecules for use in biochemical analysis.

[0019] The particles of the present disclosure may be used in a variety of homogeneous, sandwich, competitive, or non-competitive assay formats to generate a signal related to the presence or amount of an analyte in a test sample. The term "analyte," as used herein, generally refers to a substance or set of substances in a sample that is detected and / or measured, either directly or indirectly. In various aspects of the assays of the present disclosure, examples include sandwich immunoassays in which the analyte in the sample is captured between a first binding member (e.g., an antibody) attached to the particle and a second binding member for the analyte associated with a label. In another exemplary embodiment, the binding member on the particle may be an antigen (e.g., a protein) that binds an antibody of interest in a patient sample to capture the antibody on the particle. The presence of the antibody can then be detected using a label conjugated to a second binding member that is specific for the antibody. The second binding member attached to the label may be an antigen conjugated to the label, or the binding member may be an antibody (e.g., an anti-species antibody) that is itself conjugated to a label. In exemplary embodiments, these properties may be referred to herein as a "unique identifying feature" and / or "parameter" of the particle and / or of the droplet in which the particle resides. Other examples are also possible. For example, the particle may also be bound to a fluorescent tag or label, to which the fluorescent tag or label may be bound under fluorescence and / or ultraviolet emission, presenting a "unique identifying feature" and / or "parameter" of the particle.

[0020] In another exemplary embodiment, the test protocol of the present disclosure is an assay comprising a competitive immunological assay for the detection of antibodies in a sample. The competitive immunological assay may be performed in the following illustrative manner: A sample from an animal's body fluid, potentially containing an antibody of interest specific for an antigen, is contacted with the antigen attached to particles and with an anti-antigen antibody conjugated to a detectable label. The antibody of interest present in the sample competes with the antibody conjugated to a detectable label for binding to the antigen attached to the particles. The amount of label associated with the particles can then be determined after separating the unbound antibody and label. The signal obtained is inversely proportional to the amount of antibody of interest present in the sample.

[0021] In an alternative exemplary embodiment of the competitive sample, an animal's body fluid potentially containing the analyte is contacted with the analyte conjugated to a detectable label and with anti-analyte antibodies attached to particles. Antigens in the sample compete with the analyte conjugated to the label for binding to the antibodies attached to the particles. The amount of label associated with the particles can then be determined after separating the unbound antigen and label. The signal obtained is inversely proportional to the amount of analyte present in the sample.

[0022] Antibodies, antigens, and other binding members may be attached directly to the particle or to the label via covalent bonds, with or without a linker, or may be attached through distinct pairs of binding members (e.g., biotin:streptavidin, digoxigenin:antidigoxigenin) as are well known. Additionally, although the examples herein reflect use in immunological assays, the paramagnetic barcoded beads and / or particles and methods of the present disclosure may be used in other receptor binding assays, including nucleic acid hybridization assays, which rely on immobilization of one or more assay components to a solid phase.

[0023] Testing protocols, including assays, using these solutions are often performed over a series of agitation events. In practice, particles (e.g., paramagnetic barcoded beads) in solution may bind together (often referred to as "clumping"), or bind and / or settle to the bottom or sides of a container. This binding can result in inconsistent distribution of particles in solution, and if these particles clump together, they may not be accurately identified or accounted for within the testing protocol (e.g., assay).

[0024] In other examples, after one or more binding members are attached to the particles, the paramagnetic barcoded beads and / or the solution surrounding the particles may be removed from the container (e.g., cartridge), and the particles with attached binding members (collectively referred to herein as "assembled beads") may be washed in preparation for testing. In exemplary embodiments, during this wash portion, one or more components, including one or more components of the cartridge, may be used to facilitate washing, such as to fixate the assembled particles within one or more portions of the cartridge. For example, if the assembled beads have magnetic or paramagnetic properties (e.g., if they contain paramagnetic barcoded beads), a magnet may be used to fixate the assembled beads within a portion of the cartridge while the wash solution is dispersed into the cartridge, improving the results of the wash portion (e.g., by ensuring that the assembled beads remain intact and within a particular portion of the cartridge). Other improvements may also be realized.

[0025] For example, to help address these challenges, cartridges may utilize multiple electrodes that facilitate the transport of individual droplets of liquid on a surface of the cartridge. To do so, in one exemplary embodiment, the cartridge surface may comprise a dielectric material, transporting the individual droplets along one or more paths defined by multiple electrodes on a printed circuit board (PCB). Such techniques are often referred to as electrowetting on dielectrics ("EWOD"). In exemplary embodiments, the dielectric material may comprise a hydrophobic material, layer, and / or coating disposed on the surface of the PCB and / or multiple electrodes, the combination of which is referred to herein as a "dielectric cartridge surface" and / or a "path" or "paths" along the dielectric cartridge surface.

[0026] To date, such cartridges have involved a complex, interwoven series of paths along the dielectric cartridge surface. Due to several factors, including the manufacturing cost of such cartridges, there is a need to optimize cartridges that are capable of performing various steps of a test protocol but do not include components that are extraneous to the desired test protocol. Furthermore, the more complex the configuration of the cartridge and / or fluid paths on the cartridge surface, the more distance and component interactions with the fluids traveling along these paths are required. This complexity adds cost, time, and further potential errors to one or more portions of the test protocol. Therefore, in some embodiments, there is a need for a direct path for transporting droplets on the cartridge surface, including a single, generally linear path with multiple electrodes configured to transport droplets along a single path on the cartridge surface.

[0027] Disclosed herein are devices, systems, and methods for a linear path cartridge for use within EWOD systems and applications for performing one or more test protocols (e.g., assays) using one or more particles and particle types (e.g., paramagnetic barcoded beads). In exemplary embodiments, particle organization, sample analysis, and associated test protocols are improved by enabling bidirectional flow along a single, generally linear path on the surface of the cartridge.

[0028] In some embodiments, the transport of droplets on the cartridge surface can be controlled by a controller and / or other computing device to generate programmable fluidic paths that can be used in several ways (e.g., to facilitate the performance of assays and / or immunoassays). Furthermore, because the fluidic movement of droplets is controlled and programmable by a controller and / or other computing device, assay protocols and their subportions can be finely controlled to meet the needs of a desired testing protocol (e.g., assay).

[0029] In some embodiments, it is beneficial to immobilize the droplets and / or their components (e.g., paramagnetic barcoded beads) throughout one or more steps in a testing protocol (e.g., assay). In some embodiments, immobilization of the droplets on the cartridge surface can be controlled by at least one magnet. In some exemplary embodiments, the at least one magnet may be a permanent or semi-permanent magnet below or above one or more portions of the cartridge surface. In other embodiments, the at least one magnet may be an electromagnet configured, via a controller and / or other computing device, to interact with the droplets and / or their components (e.g., paramagnetic barcoded beads) and provide programmable interactions along the fluidic pathway to facilitate the assay protocol and its subportions.

[0030] In some embodiments, it is beneficial to protect or otherwise shield the droplets, their components (e.g., paramagnetic barcoded beads), and / or other materials residing on the surface of the cartridge throughout one or more steps in the assay. To do so, in some embodiments, the cartridge may be coated with one or more materials that protect the components residing on the surface of the cartridge, but still leave sufficient space on the cartridge surface for the droplets, their components (e.g., paramagnetic barcoded beads), and / or other assay components to be transported and / or immobilized on the cartridge surface. In some exemplary embodiments, this protective layer may be made from plastic and / or other materials that interact with the droplets and / or their components (e.g., paramagnetic barcoded beads), magnets, electrodes, or any other controller and / or other computing device during the assay protocol and its subportions.

[0031] In some embodiments, fluidic manipulation of droplets and their components on the cartridge surface can be facilitated by other magnets and / or electrode materials and / or controllers and other computing devices controlling them. In some embodiments, to improve fluidic transport and / or immobilization of droplets on the dielectric surface of the cartridge, one or more oils may be introduced onto the surface of the cartridge, which can be used in several ways (e.g., to improve fluidic transport of droplets during assays and / or immunoassays). Other examples are also possible.

[0032] In an exemplary embodiment, in addition to manipulating (e.g., transporting and / or immobilizing) droplets on the surface of the cartridge, various antibodies, antigens, and / or other components may also be controlled, mixed, transported, and / or immobilized on the surface of the cartridge. Using the programmable protocols, antibodies, antigens, and / or other components may be attached to the surface of one or more paramagnetic barcoded beads (“assembled beads”). In a further aspect, one or more analyses may be performed on assembled beads (or other particles) on the surface of the cartridge. In this regard, cartridge users may perform complex, often multi-step protocols, often distributed across several machines and devices within a single cartridge and a single instrument / device, at various stages of the multi-step protocol. In one exemplary embodiment, multiplexing of multiple analyte targets in a single reaction may be performed on droplets on the surface of the cartridge as detailed above, instead of using multiple devices (e.g., shaker plates, pipettes, vials, plates with multiple wells, plate readers, cameras, etc.). In one exemplary embodiment, multiplexing of multiple analyte targets in a single reaction may be performed on droplets on a portion of the surface of a cartridge that includes a single electrode.

[0033] In this regard, by combining cartridge, EWOD, magnetic, and paramagnetic barcoded bead technologies, the concepts described herein provide a disclosure for a compact, in-clinic instrument with multiplexing capabilities. By leveraging these technologies, in exemplary embodiments, a platform is described that can have the same convenience as other benchtop devices (e.g., SNAP readers and devices), but with an increased menu of capabilities for laboratory test and assay protocols, including multipart assays (e.g., multiplexed, Mpx laboratory tests), without the inconvenience and cost of the devices, instruments, and operators (e.g., liquid handling robots, plates, plate washers, and / or specialized plate readers) typically required for these tests and assays. Furthermore, in exemplary embodiments, because multiple tests and assays can be completed on one or more small sample sizes (e.g., one or more droplets containing organized paramagnetic barcoded beads), the present disclosure enables complex analyses based on small samples (e.g., of multiple analytes), which is beneficial in cases where sample volume is an issue.

[0034] In one example, a user may add a sample (e.g., a stool sample, a urine sample, a blood sample, etc.) into a reservoir of the cartridge, insert the cartridge into a benchtop instrument / device, allow the instrument / device to add and / or control other components (e.g., paramagnetic barcoded beads, solutions, antibodies, etc.) on the cartridge, analyze one or more components, and provide one or more results to a clinician, physician, and / or patient based thereon, all using the same sample, cartridge, and instrument / device. Importantly, once a user inserts the cartridge into a benchtop instrument device, some (or all) of the fluidic actions, the manipulation of the components (including the paramagnetic barcoded beads) within the cartridge, and the ultimate reading of these components are all automated, controlled, and fine-tuned by program instructions executing on a computing device, all of which may be accomplished without user interaction or control.

[0035] By doing so, several benefits are realized, including the user (e.g., clinician) having the same high-throughput / multiplexing capabilities as traditional bead technology, without the overhead required from controlling or coordinating every step of the process or the numerous separate devices and components required to perform a test and / or assay. The resulting time can also be improved; instead of shipping a sample to a laboratory and waiting for results over an extended period of time (sometimes days), the user can have results in a matter of minutes, all while using a single sample on a single cartridge associated with a single device. This improved resulting time also improves the ability of the treating physician and / or patient to receive results in a more timely manner (e.g., results can be shared with the patient during the visit) and make more timely decisions based thereon.

[0036] In a further aspect, by enabling bidirectional flow along a single, generally linear path on the cartridge surface, results from sample interaction with particles used in a testing protocol are also improved. In one example, particles (including paramagnetic barcoded beads) may be immobilized along one or more portions of the path on the cartridge surface, and a sample or samples may be transported along the path, potentially interacting with the particles more than once at different stages of the testing protocol. In this regard, sample analysis and associated testing protocols are improved as particle / sample interaction is increased. In an example, with increased particle / sample interaction, any associated particle organization and / or associated reading / imaging / analysis are also improved.

[0037] Referring now to the figures, Figure 1 is a simplified block diagram of an exemplary computing device 100 of a system (e.g., as illustrated in Figure 2, described in further detail below). Computing device 100 may perform various acts and / or functions, such as those described in this disclosure. Computing device 100 may include various components, such as a processor 102, a data storage unit 104, a communication interface 106, and / or a user interface 108. These components may be connected to each other (or to another device, system, or other entity) via a connection mechanism 110.

[0038] The processor 102 may include a general-purpose processor (eg, a microprocessor) and / or a special-purpose processor (eg, a digital signal processor (DSP)).

[0039] Data storage unit 104 may include one or more volatile, nonvolatile, removable, and / or non-removable storage components, such as magnetic, optical, or flash storage devices, and / or may be wholly or partially integrated with processor 102. Furthermore, data storage unit 104 may take the form of a non-transitory computer-readable storage medium having stored thereon program instructions (e.g., compiled or non-compiled program logic and / or machine code) that, when executed by processor 102, cause computing device 100 to perform one or more acts and / or functions, such as those described in this disclosure. Thus, computing device 100 may be configured to perform one or more acts and / or functions, such as those described in this disclosure. Such program instructions may define and / or be part of a discrete software application. In some instances, computing device 100 may execute program instructions in response to receiving input from communications interface 106 and / or user interface 108, etc. Data storage unit 104 may also store other types of data, such as those types described in this disclosure.

[0040] The communication interface 106 may enable the computing device 100 to connect to and / or communicate with other entities according to one or more protocols. In one embodiment, the communication interface 106 may be a wired interface, such as an Ethernet interface or a high-definition serial / digital interface (HD-SDI). In another embodiment, the communication interface 106 may be a wireless interface, such as a cellular or WI-FI interface. In the present disclosure, a connection may be a direct connection or an indirect connection, the latter being a connection that passes through and / or traverses one or more entities, such as routers, switches, or other network devices. Similarly, in the present disclosure, a transmission may be a direct transmission or an indirect transmission.

[0041] User interface 108, if applicable, may facilitate interaction between computing device 100 and a user of computing device 100. Thus, user interface 108 may include input components such as a keyboard, keypad, mouse, touch-sensitive panel, microphone, camera, and / or movement sensors, and / or output components such as a display device (e.g., that may be combined with a touch-sensitive panel), audio speakers, and / or a haptic feedback system, all of which may be used to obtain data indicative of the environment of computing device 100. More generally, user interface 108 may include hardware and / or software components that facilitate interaction between computing device 100 and a user of computing device 100.

[0042] The computing device 100 can take a variety of forms, such as a workstation terminal, a desktop computer, a laptop, a tablet, a mobile phone, or a controller.

[0043] 2 , a cartridge 200 is disclosed that includes a sample reservoir 202, a solution reservoir 204, an assay component reservoir 206, and a waste reservoir 208, all of which reside on a dielectric cartridge surface, according to an exemplary embodiment. In this exemplary embodiment, multiple electrodes and at least one magnet are disposed along various portions of the dielectric cartridge surface, including along a single, generally linear path 210 that extends between the sample reservoir 202 and the waste reservoir 208 and is connected to paths on the dielectric cartridge surface that lead to the solution reservoir 204 and the assay component reservoir 206. In a further aspect, the multiple electrodes and at least one magnet may be disposed under and / or around any of the illustrated sample reservoir 202, solution reservoir 204, assay component reservoir 206, waste reservoir 208, and / or portion 212.

[0044] As described above, this plurality of electrodes along the illustrated paths and reservoirs on the dielectric cartridge surface facilitates transport of fluid droplets containing multiple particles (e.g., at least one paramagnetic barcoded bead) along a single, generally linear path 210 on the dielectric cartridge surface. For clarity, as illustrated in Figure 2, the term "dielectric cartridge surface" as used in Figure 2 includes the cartridge surface below the illustrated sample reservoir 202, solution reservoir 204, assay component reservoir 206, and waste reservoir 208, as well as all of the illustrated paths connecting all of these components in Figure 2.

[0045] In examples, cartridge 200 and / or any components thereof may interact with a computing device, such as computing device 100. As described above, computing device 100 can be implemented as a controller, and a user of the controller can use the controller to program and / or control cartridge 200 and / or any components thereof. Cartridge 200 and / or any components thereof may be communicatively coupled to a controller, such as computing device 100, and may communicate with the controller via a wired connection, a wireless connection, or a combination thereof. Further, as described above, the controller may be configured to utilize cartridge 200 and / or any components thereof to control various aspects of the illustrated cartridge 200 and testing protocol (e.g., assay). While various cartridge components and arrangements of these components are provided for illustrative purposes, beads, particles, and / or components of different shapes, quantities, and / or types may also be used.

[0046] In embodiments, the controller may execute a program that causes one or more components of cartridge 200 to perform a sequence of events using a non-transitory computer-readable medium having program instructions stored thereon. These program instructions may include, for example, applying a voltage and / or current to multiple electrodes near (e.g., below) the dielectric material of the dielectric cartridge surface to transport one or more droplets (or their components) along the dielectric cartridge surface. In some embodiments, multiple electrodes may be used to transport one or more droplets between the components illustrated in FIG. 2 and / or manipulate one or more droplets and / or their components at one or more portions of the dielectric cartridge surface. For example, the controller may apply a direct current to multiple electrodes in a series of on / off voltage / current bursts, which may result in the droplets alternating between an elongated morphology on the dielectric surface (when a voltage / current is applied to the electrodes near the droplets) and a non-elongated morphology on the dielectric surface (when a voltage / current is not applied to the electrodes near the droplets). In embodiments, this oscillation of on / off voltage / current bursts and the associated morphology the droplet assumes during each event can be beneficial for mixing components within the droplet. Certain voltage / current amplitudes and patterns and electrode placement around the surface of the dielectric cartridge surface can agitate droplets more effectively than other methods, producing accurate and consistent particle mixing and associated test results.

[0047] For example, if a direct current is applied to an electrode adjacent to a droplet containing paramagnetic barcoded beads, the beads may align with the direct current and remain steady on the surface of the dielectric cartridge surface due to the paramagnetic nature of the beads. Alternatively, if an alternating current is applied to an electrode adjacent to a droplet containing paramagnetic barcoded beads, the beads may align with the direct current and alternate between two or more positions on the surface of the dielectric cartridge surface based on the alternating current due to the paramagnetic nature of the beads. It should be understood that other implementations are possible, and that complex and novel fluidic functions may be implemented due to processor-controlled fluidic actions.

[0048] For example, the controller program instructions can include moving various fluids around the surface of the cartridge to perform various aspects of a testing protocol (e.g., an assay), all on the surface of the cartridge, in an entirely automated (or primarily automated) procedure.

[0049] In this regard, in an exemplary embodiment, multiple electrodes may transport droplets containing paramagnetic barcoded beads along a single, generally linear path 210 on the dielectric cartridge surface. In examples, the paramagnetic barcoded beads may be introduced into the droplets either in liquid suspension or dried onto the surface of cartridge 200 and rehydrated. In one example, the paramagnetic barcoded beads may be suspended in a buffer solution containing sucrose, removed from suspension, and dried before being stored on portion 212 of the single, generally linear path 210 on the dielectric cartridge surface. In examples, the dried paramagnetic barcoded beads may be rehydrated with one or more solutions containing one or more components (e.g., reagents, sample, or both, among other possibilities) before being used in one or more aspects of a testing protocol (e.g., an assay). In an exemplary embodiment, once the paramagnetic barcoded beads are rehydrated and / or introduced into a fluid droplet, the droplet containing the paramagnetic barcoded beads may be transported from portion 212 to sample reservoir 202 for mixing with a sample (e.g., a stool sample, a urine sample, a blood sample, etc.) residing in the sample reservoir.

[0050] In other exemplary embodiments, dried paramagnetic barcoded beads may be stored in sample reservoir 202 and rehydrated using, among other possibilities, a solution from solution reservoir 204, a liquid accompanying the sample upon introduction into sample reservoir 202 (e.g., urine), or both.

[0051] In a further aspect, in exemplary embodiments, multiple electrodes may transport droplets of assay components (e.g., containing antibodies, antigens, labels, reagents, and / or other binding members) from assay component reservoirs 206 on the dielectric cartridge surface. In examples, these assay components may be introduced into the droplets either in liquid suspension (e.g., within component reservoirs 206, on a path approaching component reservoirs 206, or both, among other possibilities) or dried onto the surface of cartridge 200 and rehydrated. In either case, once the assay components are introduced into the droplets, the droplets containing the assay components may be transported to sample reservoir 202 for mixing with the sample and / or paramagnetic barcoded beads resident in the sample reservoir.

[0052] In other exemplary embodiments, one or more specific assay components (e.g., antibodies) may be dried and stored in sample reservoir 202, perhaps along with dried paramagnetic barcoded beads, and rehydrated in the sample reservoir using a solution from solution reservoir 204, a liquid accompanying the sample upon introduction to sample reservoir 202 (e.g., urine), or both, among other possibilities. In a further aspect, in this exemplary embodiment, one or more additional assay components (e.g., reagents, fluorescent labels / tags, etc.) may be stored in assay component reservoir 206 and transported over the dielectric cartridge surface to sample reservoir 202 for mixing with one or more specific assay components, paramagnetic barcoded beads, and / or the sample.

[0053] Furthermore, although sample reservoir 202, solution reservoir 204, and assay component reservoir 208 are illustrated as single reservoirs in Figure 2, it should be apparent to one skilled in the art that any, some, or all of these reservoirs may comprise multiple separate reservoirs, each of which may contain specific particles, components, or combinations thereof (e.g., dried paramagnetic barcoded beads and / or specific antibodies, antigens, labels, and / or other binding members). Additionally or alternatively, and specifically illustrated in Figure 2, there may be multiple assay component reservoirs within cartridge 200, each with its own associated assay component and / or pathway on the dielectric cartridge surface.

[0054] In exemplary embodiments, various techniques can be used to facilitate mixing of the sample, paramagnetic barcoded beads, and assay components within sample reservoir 202. In some examples, multiple electrodes positioned near sample reservoir 202 may be employed to circulate and / or otherwise manipulate the fluidic action of components within sample reservoir 202, including droplets containing paramagnetic barcoded beads, sample (which may contain liquid), assay components, and / or combinations thereof, among other possibilities.

[0055] For example, one or more magnets positioned near sample reservoir 202 may be employed to immobilize droplets containing paramagnetic barcoded beads, while multiple electrodes may be used in conjunction with the magnets to otherwise manipulate the fluidic behavior of other components within sample reservoir 202, including droplets containing paramagnetic barcoded beads, the sample (which may contain liquid), assay components, and / or combinations thereof, among other possibilities. For example, one or more mixing beads may also reside within sample reservoir 202, which may be controlled by magnets, electrodes, or both, and further facilitate mixing of components within sample reservoir 202 at various mixing rates, patterns, etc., all of which may be controlled by a controller executing program instructions that control the components of cartridge 200.

[0056] In an exemplary embodiment, once the droplets containing the paramagnetic barcoded beads, sample, and / or other assay components are thoroughly mixed, all of these components may be incubated (e.g., to allow attachment of one or more assay components and / or components of the sample to attach to the paramagnetic barcoded beads) in sample reservoir 202. In an exemplary embodiment, once incubation is complete, the paramagnetic barcoded beads and attached sample and / or assay components (collectively, "assembled beads") may be immobilized in sample reservoir 202 (e.g., using a magnet), while the fluid in sample reservoir 202 may be transported (e.g., using multiple electrodes) along a single, generally linear path 210 on the dielectric cartridge surface to waste reservoir 208.

[0057] After fluid is removed from sample reservoir 202, a solution (e.g., a wash solution) may be transported from solution reservoir 204 to sample reservoir 202 to wash excess residue and / or other components from the organized beads contained within sample reservoir 202. In exemplary embodiments, the solution may interact with the organized beads based on fluid action controlled by multiple electrodes that transport the solution fluid across the immobilized organized beads or through a mixing protocol with the organized beads (such as the mixing step described above). Once excess residue and / or other components have been washed from the organized beads, the solution (and any other excess fluid) may be transported from sample reservoir 202 to waste reservoir 208 along the dielectric cartridge surface (e.g., using multiple electrodes) so that the organized beads remain within sample reservoir 202 (e.g., via immobilization).

[0058] In other examples, this sequence of transporting paramagnetic barcoded beads and / or assembly components to the sample reservoir, mixing within the sample reservoir, and transporting solutions to and from the sample reservoir may be repeated several times and / or in different orders depending on the requirements of the testing protocol.

[0059] For example, a first assembly component (e.g., a detection antibody) may be stored in sample reservoir 202 and mixed with the paramagnetic barcoded beads and the sample, all of which may be allowed to incubate in sample reservoir 202. Then, after washing the first assembly component, paramagnetic barcoded beads, and sample mixture (e.g., with solution from solution reservoir 204), a second assembly component (e.g., a streptavidin reagent) stored in assay component reservoir 206 may be transported into sample reservoir 202 to complete another portion of the assembly protocol for the assembled beads before the assembled beads are analyzed.

[0060] In an example, once the assembled beads are complete and ready for analysis, they may be transported along a single, generally linear path 210 on the dielectric cartridge surface to a portion 212 of the single, generally linear path 210 for analysis. In an exemplary embodiment, the assembled beads may be transported via fluid transport from the sample reservoir 202 along the single, generally linear path 210 on the dielectric cartridge surface using multiple electrodes, via at least one magnet (i.e., moving the paramagnetic beads across the dielectric cartridge surface based on interaction with one or more magnets), or both, among other possibilities.

[0061] In examples, portion 212 of single, generally linear path 210 on the dielectric cartridge surface provides a predetermined location for a reader to perform an examination (e.g., an assay) on the assembled beads. In exemplary embodiments, the reader may detect an assay read signal corresponding to at least one of the assembled beads residing on portion 212 (potentially immobilized thereon by one or more magnets) immediately after assembly is complete. In some exemplary embodiments, this detection may occur within a predetermined time period after assembly is complete; by initiating assay read immediately after assembly, the assay provides more accurate results. In exemplary embodiments, an optical system reader may be employed to decode the individual barcodes of the paramagnetic barcoded beads. In other examples, multiple electrodes and / or one or more magnets may be used to manipulate the paramagnetic barcoded beads while reading other parameters of the droplets containing the assembled beads and / or the assembled beads themselves (e.g., by applying ultraviolet light and reading the fluorescent emission of the assembled beads via a fluorophore detector). As illustrated in FIG. 2, exploded view 214 provides an illustrative view of paramagnetic barcoded beads being read in portion 212, although it should be understood that this analysis (e.g., reading) may also occur in other portions of cartridge 200, including within sample reservoir 202.

[0062] In a further aspect, although the mixing and analysis protocols are discussed in connection with the sample reservoir 202, it should be understood that these mixing protocols may also occur in other portions of the illustrated cartridge 200, including on the portion 212 of the single, generally linear path 210 of the dielectric cartridge surface. Other implementations are also possible.

[0063] For example, a first assembly component (e.g., a detection antibody) may be stored in sample reservoir 202 and mixed with the sample in sample reservoir 202. Then, after mixing the first assembly component and the sample, the droplet of the first assembly component / sample mixture may be washed and transported along single substantially linear path 210 on the dielectric cartridge surface to portion 212 of single substantially linear path 210 and mixed with paramagnetic barcoded beads stored on portion 212. In this example, the paramagnetic barcoded beads stored on portion 212 may be mixed with the first assembly component / sample mixture (e.g., using the mixing protocol described above), and the paramagnetic barcoded beads / first assembly component / sample mixture may be allowed to incubate on portion 212. Then, after washing the paramagnetic barcoded beads / first assembly component / sample mixture (e.g., with solution from solution reservoir 204), a second assembly component (e.g., streptavidin reagent) stored in assay component reservoir 206 may be transported to portion 212 and mixed with the paramagnetic barcoded beads / first assembly component / sample mixture to complete another portion of the assembly protocol for the assembled beads before the assembled beads are analyzed. In a further aspect, in this embodiment, because mixing, incubation, washing, and / or other protocols occur in portion 212 of single generally linear path 210, specific transport of the assembled beads to this portion 212 is not required because the assembled beads are already resident in this portion 212. In an embodiment, immobilization and / or other manipulations imparted to droplets containing assembled beads on portion 212, the assembled beads themselves, and / or other components during analysis may be accomplished using multiple electrodes, at least one magnet, or both, among other possibilities.

[0064] In a further aspect, by enabling bidirectional flow along a single, generally linear path on the surface of the cartridge, as detailed in FIG. 2, sample analysis and associated testing protocols are improved based on at least the flow of components and / or sample over the beads along a single, generally linear path on the surface of the cartridge, improving time to analysis and multiple interactions between the sample and beads during assembly.

[0065] Additionally, in some exemplary embodiments, one or more of the cartridge illustrated in Figure 2, the controller illustrated in Figures 1 and 2, and / or other components of the illustrated systems may provide feedback to a user / operator via a user interface of the controller and / or benchtop device, including graphical representations of detected parameters, test results, and / or the like, to provide information to the user. Other examples are also possible.

[0066] 3 , for example, a cartridge 300 is disclosed that includes a sample reservoir 302, multiple component reservoirs 304, and a waste reservoir 306, all of which reside on a dielectric cartridge surface, according to an exemplary embodiment. In this exemplary embodiment, multiple electrodes and at least one magnet are disposed along various portions of the dielectric cartridge surface, including along a single, generally linear path 308 that extends between the sample reservoir 302 and the waste reservoir 306 and is connected to a path on the dielectric cartridge surface that leads to the component reservoir 304. In a further aspect, the multiple electrodes and at least one magnet may be disposed under and / or around any of the sample reservoir 302, component reservoir 304, waste reservoir 306, and / or portion 310 shown.

[0067] In examples, cartridge 300 and / or any components thereof may interact with a computing device, such as computing device 100, which may be implemented as a controller, which a user of the controller can use to program and / or control cartridge 300 and / or any components thereof. Additionally, as described above, the controller may be configured to control various aspects of the illustrated cartridge 300 and testing protocols (e.g., assays) utilizing cartridge 300 and / or any components thereof. While various cartridge components and arrangements of these components are provided for illustrative purposes, different shapes, quantities, and / or types of beads, particles, and / or components may also be used.

[0068] In an embodiment, the controller may execute a program that causes one or more components of cartridge 300 to perform a sequence of events using a non-transitory computer-readable medium storing program instructions, including at least those described in more detail in connection with FIG. 2 above.

[0069] In an exemplary embodiment, each, some, or all of the component reservoirs 304 may contain a solution (e.g., a buffer and / or a wash solution) that may be transported via multiple electrodes to a single, generally linear path 308 on the dielectric cartridge surface. In an example, paramagnetic barcoded beads may be contained in one of the component reservoirs 304 and introduced into a droplet, either in liquid suspension, or dried onto the surface of the cartridge 300 and rehydrated.

[0070] In one example, the paramagnetic barcoded beads may be suspended in a buffer solution containing sucrose, removed from suspension, and dried before being stored on portion 310 of single, generally linear path 308 on the dielectric cartridge surface or in a path or paths that approach one or more of component reservoirs 304. In an example, the dried paramagnetic barcoded beads may be rehydrated with one or more solutions containing one or more components (e.g., reagents, sample, or both, among other possibilities) before being used in one or more aspects of a testing protocol (e.g., an assay). In an exemplary embodiment, once the paramagnetic barcoded beads are rehydrated and / or introduced into fluidic droplets, the droplets containing the paramagnetic barcoded beads may be transported from portion 310 to sample reservoir 302 and mixed with a sample (e.g., a stool sample, a urine sample, a blood sample, etc.) residing in the sample reservoir. In other exemplary embodiments, once the paramagnetic barcoded beads are rehydrated and / or introduced into the fluid droplet, the containing droplet may be combined with one or more components stored in each, some, or all of the component reservoirs 304 and / or in a pathway approaching each, some, or all of the component reservoirs 304, and mixed in portion 310 with the paramagnetic barcoded beads resident on portion 310.

[0071] To do so, in exemplary embodiments, multiple electrodes may transport droplets of assay components (e.g., containing antibodies, antigens, labels, reagents, and / or other binding members) from one or more assay components in one or more component reservoirs 304 on the dielectric cartridge surface. In examples, these assay components may be introduced into the droplets along a single, generally linear path 308 on the dielectric cartridge surface after being rehydrated (e.g., in one or more component reservoirs 304, on one or more paths approaching the component reservoirs 304, or both, among other possibilities). In either case, once the assay components are introduced into the droplets, the droplets containing the assay components may be transported to sample reservoir 302, portion 310, and / or other portions of the single, generally linear path 308 on the dielectric cartridge surface and mixed with the sample and / or paramagnetic barcoded beads.

[0072] Additionally, while sample reservoir 302 and component reservoir 304 are illustrated as single reservoirs in Figure 3, it should be apparent to one skilled in the art that any, some, or all of these reservoirs may comprise multiple separate reservoirs, each of which may contain specific particles, components, or solutions (e.g., dried paramagnetic barcoded beads and / or specific antibodies, antigens, labels, and / or other binding members, solutions, etc.). In other exemplary embodiments, the illustrated waste reservoir 306, although illustrated in Figure 4 as being disposed on a dielectric surface of the cartridge, may be disposed on a portion of the cartridge without dielectric properties (as no fluid movement may be desired after the by-product liquid is transported to the waste reservoir).

[0073] Indeed, in the exemplary embodiment, each of the six illustrated component reservoirs 304 may contain a specific particle, component, or solution (e.g., one component reservoir contains only a wash solution, one contains only a buffer solution, one contains a first antibody, one contains a second antibody, one contains a specific reagent, and one contains a specific label). In a further aspect, any, some, or all of the specific particles, components, or solutions may be transported from the component reservoirs 304 to a single, generally linear pathway 308 on the dielectric cartridge surface via multiple electrodes, potentially after being rehydrated to hydrate on the surface. Additionally or alternatively, and particularly as illustrated in FIG. 3 , there may be more or fewer component reservoirs within cartridge 300, each with its own associated assay component and / or pathway on the dielectric cartridge surface.

[0074] As in Figure 2, in the exemplary embodiment in Figure 3, various techniques can be used to facilitate mixing of the sample, paramagnetic barcoded beads, and components in and around the various portions and reservoirs of cartridge 300, including sample reservoir 302 and portion 310. It should be understood that mixing protocols can occur in various portions of the illustrated cartridge 300 as well as various testing protocols (including assays), as discussed in detail in the context of Figure 2. As illustrated in Figure 3, exploded view 312 provides an exemplary view of paramagnetic barcoded beads being read in portion 310, and it should be understood that this analysis (e.g., reading) can also occur in other portions of cartridge 300, including in sample reservoir 302.

[0075] Additionally, in some exemplary embodiments, one or more of the cartridge illustrated in Figure 3, the controller illustrated in Figures 1 and 2, and / or other components of the illustrated systems may provide feedback to a user / operator via a user interface of the controller and / or benchtop device, including graphical representations of detected parameters, test results, and / or the like, to provide information to the user. Other examples are also possible.

[0076] For example, referring now to FIG. 4 , a cartridge 400 is disclosed that includes a sample reservoir 402, multiple component reservoirs 404, and a waste reservoir 406, all of which reside on the dielectric cartridge surface, according to an exemplary embodiment. In this exemplary embodiment, multiple electrodes and at least one magnet are disposed along various portions of the dielectric cartridge surface, including along a single, generally linear path 408 that extends between the sample reservoir 402 and a portion 410 of the single, generally linear path 408 and connects to paths on the dielectric cartridge surface that lead to the component reservoirs 404, all of which terminate in the waste reservoir 406. Unlike FIGS. 2 and 3 , in FIG. 4 , the waste reservoir 406 does not reside at the end of the single, generally linear path 408. Thus, when fluids and / or other components are transported to the waste reservoir 406, they are not transported exclusively via the single, generally linear path 408. Instead, these waste products may be transported to a waste reservoir via any one of the illustrated paths of the plurality of illustrated component reservoirs 404. In a further aspect, a plurality of electrodes and at least one magnet may be disposed under and / or around any of the illustrated sample reservoirs 402, component reservoirs 404, waste reservoirs 406, and / or portions 410.

[0077] In embodiments, cartridge 400 and / or any of its components may interact with a computing device, which may be implemented as a controller, which a user of the controller can use to program and / or control cartridge 400 and / or any of its components. Additionally, as described above, the controller may be configured to control various aspects of the illustrated cartridge 400 and testing protocols (e.g., assays) utilizing cartridge 400 and / or any of its components. While various cartridge components and arrangements of these components are provided for illustrative purposes, different shapes, quantities, and / or types of beads, particles, and / or components may also be used.

[0078] In an embodiment, the controller may execute a program that causes one or more components of cartridge 400 to perform a sequence of events using a non-transitory computer-readable medium storing program instructions, including at least those described in further detail in connection with Figures 2 and 3 above.

[0079] In an exemplary embodiment, each, some, or all of the component reservoirs 404 may contain a solution (e.g., a buffer and / or a wash solution) that may be transported via multiple electrodes to a single, generally linear path 408 on the dielectric cartridge surface. In an example, paramagnetic barcoded beads may be contained in one of the component reservoirs 404 and introduced into a droplet, either in liquid suspension, or dried onto the surface of the cartridge 400 and rehydrated.

[0080] In one example, the paramagnetic barcoded beads may be suspended in a buffer solution containing sucrose, removed from suspension, and dried before being stored on portion 410 of single generally linear path 408 on the dielectric cartridge surface or in a path or paths that approach one or more of component reservoirs 404. In an example, the dried paramagnetic barcoded beads may be rehydrated with one or more solutions containing one or more components (e.g., reagents, sample, or both, among other possibilities) before being used in one or more aspects of a testing protocol (e.g., an assay).

[0081] In an exemplary embodiment, once the paramagnetic barcoded beads have been rehydrated and / or introduced into the fluid droplet, the droplet containing the paramagnetic barcoded beads may be transported from portion 410 to sample reservoir 402 and mixed with a sample residing in the sample reservoir (e.g., a stool sample, a urine sample, a blood sample, etc.).

[0082] In other exemplary embodiments, once the paramagnetic barcoded beads are rehydrated and / or introduced into the fluid droplet, the containing droplet may be combined with one or more components stored in each, some, or all of the component reservoirs 404 and / or in a pathway approaching each, some, or all of the component reservoirs 404, and mixed in portion 410 with the paramagnetic barcoded beads resident on portion 410.

[0083] To do so, in exemplary embodiments, multiple electrodes may transport droplets of assay components (e.g., containing antibodies, antigens, labels, reagents, and / or other binding members) from one or more assay components stored in one or more component reservoirs 404 on the dielectric cartridge surface. In examples, these assay components may be introduced into the droplets along a single, generally linear path 408 on the dielectric cartridge surface after being rehydrated (e.g., in one or more component reservoirs 404, on one or more paths approaching the component reservoirs 404, or both, among other possibilities). In either case, once the assay components are introduced into the droplets, the droplets containing the components (e.g., assay components) or some combination thereof may be transported to sample reservoir 402, portion 410, and / or other portions of the single, generally linear path 408 on the dielectric cartridge surface and mixed with the sample, paramagnetic barcoded beads, and / or components, among other possibilities.

[0084] Additionally, while sample reservoir 402 and component reservoir 404 are depicted as single reservoirs in Figure 4, it should be apparent to one skilled in the art that any, some, or all of these reservoirs may comprise multiple separate reservoirs, each of which may contain specific particles, components, or solutions (e.g., dried paramagnetic barcoded beads and / or specific antibodies, antigens, labels, and / or other binding members, solutions, etc.). In other exemplary embodiments, although the depicted waste reservoir 406 is depicted in Figure 4 as being disposed on a dielectric surface of the cartridge, this reservoir may be located on a portion of the cartridge without dielectric properties (as no fluid movement may be desired after the by-product liquid is transported to the waste reservoir).

[0085] 2 and 3, in exemplary embodiments, various techniques can be used to facilitate mixing of sample, paramagnetic barcoded beads, and components in and around the various portions and reservoirs of cartridge 300, including sample reservoir 302 and portion 310. It should be understood that mixing protocols can occur in various portions of the illustrated cartridge 400 and various testing protocols (including assays), as discussed in detail in the context of FIGS. 2 and 3. As illustrated in FIG. 4, exploded view 412 provides an exemplary view of paramagnetic barcoded beads being read in portion 410, and it should be understood that this analysis (e.g., reading) can also occur in other portions of cartridge 400, including within sample reservoir 402.

[0086] Additionally, in some exemplary embodiments, one or more of the cartridge illustrated in Figure 4, the controller illustrated in Figures 1-3, and / or other components of the illustrated systems may provide feedback to a user / operator via a user interface of the controller and / or benchtop device, including graphical representations of detected parameters, test results, and / or the like, to provide information to the user. Other examples are also possible.

[0087] Exemplary Methods and Aspects Referring now to FIG. 5, an exemplary method for analyzing droplets on a surface of a cartridge, where the droplets include a plurality of particles, is disclosed.

[0088] The method 500 shown in Figure 5 presents one example of a method that may be used in conjunction with the components shown in Figures 1-4, for example. Furthermore, a device or system may be used or configured to perform the logical functions presented in Figure 5. In other examples, the components of the device and / or system may be arranged such that they are adapted, capable of, or suitable for performing a function, such as when operated in a specific manner. The method 500 may include one or more operations, functions, or actions, as illustrated by one or more of blocks 502-504. Although the blocks are illustrated in a sequential order, these blocks may also be performed in parallel and / or in a different order than described herein.

[0089] Additionally, various blocks may be combined into fewer blocks, divided into additional blocks, and / or eliminated based on the desired implementation.

[0090] In block 502, a method 500 for analyzing droplets on a surface of a cartridge, where the droplets include multiple particles, involves transporting the droplets along a single path on the surface of the cartridge via multiple electrodes of the cartridge, the single path being approximately linear, and the multiple electrodes being configured to transport the droplets along the single path on the surface of the cartridge.

[0091] In some exemplary embodiments, the at least one paramagnetic barcoded bead of the droplet comprises at least one paramagnetic barcoded bead. In some exemplary embodiments, the at least one paramagnetic barcoded bead of the droplet comprises one or more unique barcodes. In other examples, the at least one paramagnetic barcoded bead of the droplet comprises at least one non-spherical paramagnetic barcoded bead. In some examples, the at least one paramagnetic barcoded bead of the droplet is between about 0.1 and 100 microns in size. In some examples, the droplet further comprises a solution for washing the plurality of particles of the droplet. In some examples, the droplet further comprises a Read buffer solution.

[0092] In some embodiments, the single path on the surface of the cartridge comprises a dielectric material, and transporting the droplet along the single path on the surface of the cartridge via the plurality of electrodes of the cartridge comprises applying a current to the electrodes of the cartridge. In some embodiments, the current comprises a direct current. In some embodiments, the current comprises an alternating current.

[0093] In some embodiments, the single channel on the surface of the cartridge further comprises a sample reservoir and a waste reservoir.

[0094] At block 504, the method 500 involves analyzing the droplets at one or more locations along a single path on the surface of the cartridge.

[0095] In an embodiment, analyzing droplets at one or more locations along a single path on the surface of the cartridge includes analyzing droplets in a sample reservoir.

[0096] In some embodiments, method 500 further involves immobilizing the droplets at specific locations along a single path on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets along the single path on the surface of the cartridge. In some embodiments, immobilizing the droplets at specific locations along the single path on the surface of the cartridge via the at least one magnet of the cartridge includes immobilizing at least one paramagnetic barcoded bead of the droplets. In some embodiments, analyzing the droplets at one or more locations along the single path on the surface of the cartridge includes analyzing the droplets while they are immobilized at the specific locations along the single path on the surface of the cartridge.

[0097] Additionally, in some embodiments, analyzing the droplets at one or more locations along the single path on the surface of the cartridge includes performing one or more assay procedures on the droplets at one or more locations along the single path on the surface of the cartridge, and determining parameters of the droplets during the one or more assay procedures. In some embodiments, determining the parameters of the droplets includes identifying a particular characteristic of the plurality of particles.

[0098] In some embodiments, analyzing the droplets at one or more locations along a single path on the surface of the cartridge includes generating an image of the droplet at one or more locations along a single path on the surface of the cartridge, the image including an image of a plurality of particles, and determining parameters of the droplets based on the generated image. In some embodiments, determining parameters of the droplets includes comparing the generated image of the droplet with a previously generated image of the droplet. In some embodiments, analyzing the droplets further includes applying ultraviolet light to the droplets while generating the image of the droplets at one or more locations along the single path on the surface of the cartridge. In some embodiments, analyzing the droplets includes generating a composite image of the droplets on the surface of the cartridge, the composite image including a plurality of images of at least one paramagnetic barcoded bead in the droplet, and determining parameters of the droplets based on the generated composite image. In some embodiments, method 500 includes transmitting instructions that cause a graphical user interface to display a graphical representation of the determined parameters of the droplets.

[0099] In some embodiments, analyzing the droplets at one or more locations along a single path on the surface of the cartridge includes performing multiple assay procedures on the droplets at one or more locations along the single path on the surface of the cartridge, and determining the presence of one or more analytes attached to at least one paramagnetic barcoded bead of the droplets during the one or more assay procedures.

[0100] In one aspect, a non-transitory computer-readable medium is disclosed having program instructions stored thereon, which, upon execution by a controller, cause the controller to perform a set of operations including transporting a droplet along a single path on a surface of the cartridge via a plurality of electrodes of the cartridge, the droplet including a plurality of particles, the single path being generally linear, and the plurality of electrodes configured to transport the droplet along the single path on the surface of the cartridge; and analyzing the droplet at one or more locations along the single path on the surface of the cartridge.

[0101] The singular articles "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0102] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will surely become apparent to those skilled in the art. Additionally, the various aspects and embodiments disclosed herein are provided for illustrative purposes and are not intended to be limiting, with the true scope being indicated by the following claims.

Claims

1. 1. A method for analyzing a droplet on a surface of a cartridge, the droplet including a plurality of particles, the method comprising: transporting the droplet along a single path on the surface of the cartridge via a plurality of electrodes of the cartridge, the single path being substantially linear, and the plurality of electrodes configured to transport the droplet along the single path on the surface of the cartridge; analyzing the droplets at one or more locations along the single path on the surface of the cartridge; A method comprising:

2. 10. The method of claim 1, wherein the plurality of particles comprises at least one paramagnetic barcoded bead.

3. 3. The method of claim 2, wherein the at least one paramagnetic barcoded bead comprises one or more unique barcodes.

4. 3. The method of claim 2, wherein the at least one paramagnetic barcoded bead comprises at least one non-spherical paramagnetic barcoded bead.

5. 3. The method of claim 2, wherein the at least one paramagnetic barcoded bead is about 0.1 to 100 microns in size.

6. 2. The method of claim 1, wherein a single path on the surface of the cartridge comprises a dielectric material, and transporting the droplet on the surface of the cartridge via a plurality of electrodes of the cartridge comprises applying a current to the electrodes of the cartridge.

7. The method of claim 6 , wherein the current comprises a direct current.

8. The method of claim 6 , wherein the current comprises an alternating current.

9. The method of claim 1 , wherein the single channel on the surface of the cartridge further comprises a sample reservoir and a waste reservoir.

10. 10. The method of claim 9, wherein analyzing the droplets at one or more locations along the single path on the surface of the cartridge comprises analyzing the droplets in the sample reservoir.

11. 10. The method of claim 1, further comprising immobilizing the droplets at specific locations along the single path on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets along the single path on the surface of the cartridge.

12. 12. The method of claim 11, wherein immobilizing the droplet at a specific location along the single path on the surface of the cartridge via at least one magnet of the cartridge comprises immobilizing the at least one paramagnetic barcoded bead of the droplet.

13. 12. The method of claim 11, wherein analyzing the droplets at one or more locations along the single path on the surface of the cartridge comprises analyzing the droplets while they are immobilized at specific locations along the single path on the surface of the cartridge.

14. 12. The method of claim 11 , further comprising transporting additional droplets along the single path, the additional droplets comprising at least one of a sample and a reagent, and the additional droplets interacting with the immobilized droplets.

15. 15. The method of claim 14, wherein the additional droplets are transported along the single path such that the additional droplets interact with the immobilized droplets more than once.

16. 16. The method of claim 15, wherein during analyzing the droplet at one or more locations along the single path on the surface of the cartridge, an increased analysis signal is produced due to the two or more interactions between the droplet and at least one of the sample and the reagent of the additional droplet.

17. 12. The method of claim 11, wherein immobilizing the droplet at a specific location along the single path on the surface of the cartridge via at least one magnet of the cartridge comprises immobilizing the at least one paramagnetic barcoded bead of the droplet, and the method further comprises transporting fluid contained in the droplet to a waste reservoir.

18. 20. The method of claim 17, further comprising transporting additional droplets along the single path, the additional droplets comprising at least one of a sample and a reagent, the additional droplets interacting with the immobilized at least one paramagnetic barcoded bead.

19. 20. The method of claim 18, wherein the additional droplets are transported along the single path such that the additional droplets interact with the immobilized at least one paramagnetic barcoded bead more than once.

20. 20. The method of claim 19, wherein the method further comprises analyzing the at least one paramagnetic barcoded bead, and during analyzing the at least one paramagnetic barcoded bead at one or more locations along the single path on the surface of the cartridge, an increased analysis signal is produced due to the two or more interactions between the at least one paramagnetic barcoded bead and at least one of the sample and the reagent of the additional droplet.

21. 2. The method of claim 1, wherein analyzing the droplets at one or more locations along the single path on the surface of the cartridge comprises performing one or more assay procedures on the droplets at the one or more locations along the single path on the surface of the cartridge, and determining parameters of the droplets during the one or more assay procedures.

22. The method of claim 21 , wherein determining the parameters of the droplets comprises identifying particular characteristics of the plurality of particles.

23. analyzing the droplets at one or more locations along the single path on the surface of the cartridge includes generating an image of the droplets at the one or more locations along the single path on the surface of the cartridge, the image including an image of the plurality of particles; determining parameters of the droplet based on the generated image; 23. The method of claim 22, comprising:

24. 24. The method of claim 23, wherein determining the parameters of the droplet comprises comparing the generated image of the droplet with a previously generated image of the droplet.

25. 24. The method of claim 23, wherein analyzing the droplet further comprises applying ultraviolet light to the droplet while generating an image of the droplet at the one or more locations along the single path on the surface of the cartridge.

26. 22. The method of claim 21, wherein the method further comprises transmitting instructions that cause a graphical user interface to display a graphical representation of the determined parameters of the droplet.

27. 2. The method of claim 1, wherein analyzing the droplets at one or more locations along the single path on the surface of the cartridge comprises performing a plurality of assay procedures on the droplets at the one or more locations along the single path on the surface of the cartridge, and determining the presence of one or more analytes attached to the at least one paramagnetic barcoded bead of the droplets during the plurality of assay procedures.

28. The method of claim 1 , wherein the droplet further comprises a solution for washing the plurality of particles of the droplet.

29. The method of claim 1 , wherein the droplet further comprises a Reed buffer solution.

30. A non-transitory computer-readable medium having program instructions stored thereon, the program instructions, upon execution by a controller, causing the controller to: transporting a droplet along a single path on a surface of the cartridge via a plurality of electrodes of the cartridge, the droplet including a plurality of particles, the single path being substantially linear, and the plurality of electrodes configured to transport the droplet along the single path on the surface of the cartridge; analyzing the droplets at one or more locations along the single path on the surface of the cartridge; 10. A non-transitory computer-readable medium for causing a set of operations to be performed, the set including:

31. A cartridge, A plurality of electrodes; a single path for transporting droplets on a surface of the cartridge, the single path being substantially linear, the plurality of electrodes being configured to transport droplets along the single path on the surface of the cartridge, the droplets including a plurality of particles; and one or more locations for analyzing the droplets along the single path on the surface of the cartridge; A cartridge comprising:

Images