Methods, systems, and devices for electrowetting barcoded magnetic beads

Abstract

A method for manipulating droplets on a surface of a cartridge is disclosed, the droplets comprising at least one paramagnetic barcoded bead. The method includes transporting the droplets on the surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplets on the surface of the cartridge. The method also includes immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet being configured to immobilize the droplets on the surface of the cartridge.

Description

[Technical Field]

[0001] FIELD OF THE DISCLOSURE The present disclosure involves systems and methods for analyzing droplets that include at least one paramagnetic bead on a surface of a cartridge, i.e., the disclosed devices and methods transport and / or immobilize the droplets on the surface of the 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 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 paramagnetic beads and other components in droplets of solution containing the sample to aid in conducting the assay. Summary of the Invention [Means for solving the problem]

[0003] (summary) In some examples, a plurality of particles (including 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 solution). To increase the accuracy of assay test results, it is desirable to ensure that the paramagnetic barcoded beads are dispersed throughout the solution prior to testing and are properly manipulated (e.g., transported and / or immobilized) during mixing of the components of the assay and during reading of the resulting solution.

[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 the operator allows an excessive amount of time to pass between manually 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 assay results in 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 assay results.

[0005] In one embodiment, a method for manipulating droplets of a solution on a surface of a cartridge is described, the droplets comprising at least one paramagnetic barcoded bead, the method including transporting the droplets on the surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplets on the surface of the cartridge, and immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet being configured to immobilize the droplets 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 droplets on a surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes configured to transport the droplets on the surface of the cartridge, and immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets on the surface of the cartridge.

[0007] The features, functions, and advantages discussed can be achieved independently in various embodiments and may be combined in still 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 and additional features will be better understood through the following illustrative and non-limiting detailed description of illustrative 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 method according to an example embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0012] 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.

[0013] (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.

[0014] 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.

[0015] 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. The paramagnetic barcoded beads, particles, or both 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.

[0016] 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 may be modified for one or more specific uses. For example, one or more types of paramagnetic barcoded beads, one or more types of particles, and / or both 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. For example, one or more types of paramagnetic barcoded beads, one or more types of particles, and / or both particles may be surface-modified and / or functionalized with biomolecules for use in biochemical analysis.

[0017] The paramagnetic barcoded beads and / or 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 paramagnetic barcoded beads and / or particles and a second binding member for the analyte associated with the identifier. In another exemplary embodiment, the binding member on the paramagnetic barcoded beads and / or particles 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 the label. In exemplary embodiments, these properties may be referred to herein as a "unique identifying feature" and / or "parameter" of the paramagnetic barcoded beads and / or particles and / or of the droplets in which the paramagnetic barcoded beads and / or particles reside. Other examples are also possible. For example, the paramagnetic barcoded beads and / or particles may also be bound to a fluorescent tag or label, which may present a "unique identifying feature" and / or "parameter" of the paramagnetic barcoded beads and / or particles, to which the fluorescent tag or label may bind under fluorescence and / or ultraviolet emission.

[0018] 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 exemplary 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 paramagnetic barcoded beads and / or 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 paramagnetic barcoded beads and / or particles. The amount of label associated with the paramagnetic barcoded beads and / or 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.

[0019] 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 an anti-analyte antibody attached to paramagnetic barcoded beads and / or particles. Antigens in the sample compete with the analyte conjugated to the label for binding to the antibody bound to the paramagnetic barcoded beads and / or particles. The amount of label associated with the paramagnetic barcoded beads and / or 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.

[0020] Antibodies, antigens, and other binding members may be attached directly to the paramagnetic barcoded beads and / or particles via covalent bonds, with or without a linker, or to a label, or may be attached through distinct pairs of binding members (e.g., biotin:streptavidin, digoxigenin:antidigoxigenin) as are well known. In addition, 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.

[0021] Testing protocols, including assays, using these solutions are often performed over a series of agitation events. In practice, paramagnetic barcoded beads and / or particles 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 the paramagnetic barcoded beads and / or particles in the solution. When these paramagnetic barcoded beads and / or particles clump together, they may not be accurately identified or accounted for within the testing protocol (e.g., assay).

[0022] In other examples, after one or more binding members are attached to the paramagnetic barcoded beads and / or particles, the solution surrounding the paramagnetic barcoded beads and / or particles may be removed from the container (e.g., cartridge), and the paramagnetic barcoded beads and / or particles with the 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.

[0023] 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 to transport 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."

[0024] 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).

[0025] 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 the 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.

[0026] 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.

[0027] 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.

[0028] 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.

[0029] 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 multi-part 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.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] The processor 102 may include a general-purpose processor (eg, a microprocessor) and / or a special-purpose processor (eg, a digital signal processor (DSP)).

[0034] 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.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 2 , according to an exemplary embodiment, a cartridge 200 is disclosed that includes a sample reservoir 202, a solution reservoir 204, a paramagnetic barcoded bead reservoir 206, an assay component reservoir 208, a test reservoir 210, and a waste reservoir 212, all of which reside on a dielectric cartridge surface 214. In this exemplary embodiment, a plurality of electrodes and at least one magnet are disposed along various portions of the dielectric cartridge surface 214. As described above, the plurality of electrodes facilitates transport of fluidic droplets containing at least one paramagnetic barcoded bead along the dielectric cartridge surface 214, and the at least one magnet immobilizes the fluidic droplets and / or their components (e.g., at least one paramagnetic barcoded bead). For clarity, as illustrated in FIG. 2, the term "dielectric cartridge surface 214" as used in FIG. 2 includes the cartridge surface below the illustrated sample reservoir 202, solution reservoir 204, paramagnetic barcoded bead reservoir 206, assay component reservoir 208, test reservoir 210, and waste reservoir 212, as well as the illustrated pathways connecting all of these components in FIG. 2.

[0039] 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 protocols (e.g., assays). 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.

[0040] In embodiments, the controller may execute a program, using a non-transitory computer-readable medium having program instructions stored thereon, that causes one or more components of cartridge 200 to perform a sequence of events. These program instructions may include, for example, applying a voltage and / or current to multiple electrodes near (e.g., below) the dielectric material of dielectric cartridge surface 214 to manipulate one or more droplets (or their components) along dielectric cartridge surface 214. 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 dielectric cartridge surface 214. 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 configuration on the dielectric surface (when a voltage / current is applied to the electrodes near the droplets) and a non-elongated configuration on the dielectric surface (when a voltage / current is not applied to the electrodes near the droplets). 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 214 can agitate the droplets more effectively than other methods, producing accurate and consistent mixing of paramagnetic barcoded beads and particles and associated assay results. For example, if a direct current is applied to electrodes adjacent to droplets containing paramagnetic barcoded beads, the beads may align with the direct current and remain steady on the surface of the dielectric cartridge surface 214 due to the paramagnetic properties of the beads. Alternatively, if an alternating current is applied to electrodes adjacent to droplets 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 214 based on the alternating current due to the paramagnetic properties of the beads. It should be understood that other implementations are possible, and that complex and novel fluidic functions can be implemented due to processor-controlled fluidic actions.

[0041] 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.

[0042] In this regard, in an exemplary embodiment, a plurality of electrodes may transport droplets containing paramagnetic barcoded beads on the dielectric cartridge surface 214. In examples, the paramagnetic barcoded beads may be introduced into the droplets either in liquid suspension or dried onto the surface of the cartridge 200 and rehydrated. In one example, the paramagnetic barcoded beads may be suspended in a buffer solution containing sucrose and dried before being removed from suspension and stored in the paramagnetic barcoded bead reservoir 206. 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 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 the paramagnetic barcoded bead reservoir 206 to the 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.

[0043] 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.

[0044] In a further aspect, in an exemplary embodiment, a plurality of electrodes may transport droplets of assay components (e.g., containing antibodies, antigens, labels, and / or other binding members) on the dielectric cartridge surface 214. In examples, these assay components may be introduced into the droplets either in liquid suspension or dried onto the surface of cartridge 200 and rehydrated. In either case, once the paramagnetic barcoded beads are introduced into the droplets, the droplets containing the paramagnetic barcoded beads may be transported from paramagnetic assay component reservoir 208 to sample reservoir 202 for mixing with the sample residing in the sample reservoir.

[0045] 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 into 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 208 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.

[0046] Further, although assay component reservoir 208 is illustrated as a single reservoir in Figure 2, it should be apparent to one skilled in the art that assay component reservoir 208 may comprise multiple separate reservoirs, each of which may contain a particular assay component or combination thereof (e.g., a particular antibody, antigen, label, and / or other binding member). 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 dielectric cartridge surface 214.

[0047] 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.

[0048] 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.

[0049] 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 along dielectric cartridge surface 214 to waste reservoir 212 (e.g., using multiple electrodes).

[0050] 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 212 along dielectric cartridge surface 214 (e.g., using multiple electrodes) so that the organized beads remain within sample reservoir 202 (e.g., via immobilization).

[0051] In other examples, this sequence of transporting paramagnetic barcoded beads and / or assembly components to the sample reservoir, mixing in the sample reservoir, and transporting solutions to and from the sample reservoir may be repeated several times. Additionally, or alternatively, one or more assembly components may be transported to be mixed with the paramagnetic barcoded beads and / or other assembly components after a step of the assay is completed. For example, a first assembly component (e.g., an antibody) may be mixed with the paramagnetic barcoded beads and sample, and then, after washing, a second assembly component may be transported into sample reservoir 202 (e.g., a label) to complete another portion of the assembly protocol for the assembly beads.

[0052] In a further aspect, although the mixing 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 the dielectric cartridge surface portion 216. Other embodiments are also possible.

[0053] In an example, once the organized beads are complete and ready for analysis, they may be transported to a portion of the cartridge for analysis, including test reservoir 210. In an exemplary embodiment, the organized beads may be transported via fluid transport across dielectric cartridge surface 214, via magnetic forces (i.e., moving paramagnetic beads across dielectric cartridge surface 214 based on interaction with one or more magnets), or both, among other possibilities.

[0054] In examples, the test reservoir 210 provides a predetermined location for a reader to perform assay tests on the assembled beads. In exemplary embodiments, the reader may detect an assay read signal corresponding to at least one of the assembled beads in the test reservoir 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 droplet 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, the exploded view 218 provides an exemplary view of the paramagnetic barcoded beads, and it should be understood that this analysis (e.g., reading) may also occur in other portions of the cartridge 200, including the dielectric cartridge surface portion 216.

[0055] Additionally, in some exemplary embodiments, one or more components of the shaker plate system may provide feedback to the user / operator, including graphical representations of detected parameters, test results, and / or the like, via a user interface on the controller and / or tabletop device to provide information to the user.

[0056] Exemplary Methods and Aspects Referring now to FIG. 3, an exemplary method of manipulating droplets on a surface of a cartridge is shown, where the droplets include at least one paramagnetic barcoded bead.

[0057] The method 300 shown in Figure 3 presents one example of a method that may be used in conjunction with the components shown in Figures 1-2, for example. Furthermore, a device or system may be used or configured to perform the logical functions presented in Figure 3. In other examples, the components of the device and / or system may be arranged such that they are adapted, capable, or suitable to perform a function, such as when operated in a specific manner. The method 300 may include one or more operations, functions, or actions, as illustrated by one or more of blocks 302-304. 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.

[0058] Additionally, various blocks may be combined into fewer blocks, divided into additional blocks, and / or eliminated based on the desired implementation.

[0059] In block 302, a method 300 for manipulating a droplet on a surface of a cartridge, where the droplet includes at least one paramagnetic barcoded bead, involves transporting the droplet on the surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplet on the surface of the cartridge.

[0060] In some exemplary embodiments, at least one paramagnetic barcoded bead of the droplet comprises one or more unique barcodes. In other examples, at least one paramagnetic barcoded bead of the droplet comprises at least one non-spherical paramagnetic barcoded bead. In some examples, 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 at least one paramagnetic barcoded bead of the droplet. In some examples, the droplet further comprises a Read buffer solution.

[0061] In some embodiments, the surface of the cartridge comprises a dielectric material, and transporting the droplets on the surface of the cartridge via a 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.

[0062] At block 304, the method 300 involves immobilizing the droplets on a surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets on the surface of the cartridge.

[0063] In an embodiment, immobilizing the droplet on the surface of the cartridge via at least one magnet of the cartridge comprises immobilizing at least one paramagnetic barcoded bead of the droplet.

[0064] Additionally, in some embodiments, method 300 further includes analyzing the droplets while they are transported or immobilized on the surface of the cartridge. In some embodiments, analyzing the droplets includes performing one or more assay procedures on the droplets and determining parameters of the droplets during the one or more assay procedures. In other embodiments, determining parameters of the droplets includes identifying a particular characteristic of at least one paramagnetic barcoded bead in the droplets.

[0065] In some embodiments, analyzing the droplet includes generating an image of the droplet on the surface of the cartridge, the image including an image of at least one paramagnetic barcoded bead in the droplet, and determining parameters of the droplet based on the generated image. In some embodiments, determining parameters of the droplet includes comparing the generated image of the droplet with a previously generated image of the droplet. In some embodiments, analyzing the droplet further includes applying ultraviolet light to the droplet while generating the image of the droplet on the surface of the cartridge. In some embodiments, analyzing the droplet includes generating a composite image of the droplet on the surface of the cartridge, the composite image including a plurality of images of the at least one paramagnetic barcoded bead in the droplet, and determining parameters of the droplet based on the generated composite image. In some embodiments, method 300 includes transmitting instructions that cause a graphical user interface to display a graphical representation of the determined parameters of the droplet.

[0066] In one aspect, a non-transitory computer-readable medium is disclosed having program instructions stored thereon, the program instructions, when executed by a controller, causing the controller to perform a set of operations including transporting droplets on a surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplets on the surface of the cartridge, and immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet being configured to immobilize the droplets on the surface of the cartridge.

[0067] 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.

[0068] 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 manipulating droplets on a surface of a cartridge, said droplets comprising at least one paramagnetic barcoded bead, said method comprising: transporting the droplet on the surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplet on the surface of the cartridge; and immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets on the surface of the cartridge; A method comprising:

2. 10. The method of claim 1, wherein the at least one paramagnetic barcoded bead of the droplet comprises one or more unique barcodes.

3. 10. The method of claim 1, wherein the at least one paramagnetic barcoded bead of the droplet comprises at least one non-spherical paramagnetic barcoded bead.

4. 10. The method of claim 1, wherein the at least one paramagnetic barcoded bead of the droplet is about 0.1 to 100 microns in size.

5. 2. The method of claim 1, wherein 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.

6. The method of claim 5 , wherein the current comprises a direct current.

7. The method of claim 5 , wherein the current comprises an alternating current.

8. 2. The method of claim 1, wherein immobilizing the droplet 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.

9. The method of claim 1 , further comprising analyzing the droplets while they are transported or immobilized on the surface of the cartridge.

10. analyzing the droplets performing one or more assay procedures on said droplets; determining parameters of the droplets during the one or more assay procedures; 10. The method of claim 9, comprising:

11. 11. The method of claim 10, wherein determining a parameter of the droplet comprises identifying a particular characteristic of the at least one paramagnetic barcoded bead of the droplet.

12. analyzing the droplets generating an image of the droplet on the surface of the cartridge, the image including an image of the at least one paramagnetic barcoded bead in the droplet; determining parameters of the droplet based on the generated image; 10. The method of claim 9, comprising:

13. The method of claim 12 , wherein determining parameters of the droplet comprises comparing the generated image of the droplet with a previously generated image of the droplet.

14. The method of claim 12 , wherein analyzing the droplet further comprises applying ultraviolet light to the droplet while generating an image of the droplet on the surface of the cartridge.

15. analyzing the droplets performing a plurality of assay procedures on said droplets; determining the presence of one or more analytes attached to the at least one paramagnetic barcoded bead of the droplet during the one or more assay procedures; and 10. The method of claim 9, comprising:

16. The method of claim 12 , 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.

17. analyzing the droplets generating a composite image of the droplet on the surface of the cartridge, the composite image including a plurality of images of the at least one paramagnetic barcoded bead of the droplet; determining parameters of the droplet based on the generated composite image; 10. The method of claim 9, comprising:

18. The method of claim 1 , wherein the droplet further comprises a solution for washing the at least one paramagnetic barcoded bead of the droplet.

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

20. 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 on a surface of the cartridge via a plurality of electrodes of the cartridge, the plurality of electrodes being configured to transport the droplet on the surface of the cartridge; immobilizing the droplets on the surface of the cartridge via at least one magnet of the cartridge, the at least one magnet configured to immobilize the droplets 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:

21. A cartridge, A plurality of electrodes; at least one magnet; a first surface for transporting a droplet on the first surface of the cartridge via the plurality of electrodes, the droplet comprising at least one paramagnetic barcoded bead, and the plurality of electrodes configured to transport the droplet on the first surface of the cartridge; a second surface for immobilizing the droplets on the second surface via the at least one magnet, the at least one magnet configured to immobilize the droplets on the second surface of the cartridge; and A cartridge comprising:

22. 22. The cartridge of claim 21, wherein the first surface of the cartridge and the second surface of the cartridge are the same surface.

23. 22. The cartridge of claim 21, wherein the first surface of the cartridge and the second surface of the cartridge are different surfaces.

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