Systems and methods for dynamic force spectroscopy in drug discovery

Abstract

Described are systems and methods for identifying an analyte in drug discovery and development. A method can include directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; using the capture moiety to capture the analyte; applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and using the magnetic field condition to identify at least one event of the analyte, wherein the magnetic field condition comprises a time-varying force, at least one period of time, at least one motion path, or any combination thereof.

Description

SYSTEMS AND METHODS FOR DYNAMIC FORCE SPECTROSCOPYIN DRUG DISCOVERYCROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 664,816, filed June 27, 2024, of which application is incorporated herein by reference in its entirety.BACKGROUND

[0002] Determining molecular affinity can be important in the study of biological sciences to practically influence biological processes. Scientists have used molecular affinity in different applications, e.g., determining drug targets in drug discovery. Some tools used to help understand the pharmacology of drugs, e.g., immunoassays, DNA sequencing, or DNA microarrays, may rely on molecular affinity. However, some methods for understanding molecular affinity are deficient because they may not be convenient to use, may not obtain high-resolution data or even sufficient data, or may not use high-throughput methods for obtaining the data.SUMMARY

[0003] Recognized herein is a need for systems and methods that can determine molecular affinity using high-throughput methods that are easier to use and yet can provide higher-resolution, sufficient data. Such systems and methods can be applied to drug discovery and development for improving healthcare outcomes.

[0004] In an aspect, disclosed herein is a method for identifying an analyte, the method comprising: (a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; (b) using the capture moiety to capture the analyte; (c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and (d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a timevarying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii). In some embodiments, the at least one event comprises a dissociation event between the capture moiety and the analyte. In some embodiments, the method further comprises dynamically updating the magnetic field condition in real time based at least on determining a statistical quality of data for each of the events. In some embodiments, updating the magnetic field condition comprises changing at least one parameter of the time-varying force. In someembodiments, updating the magnetic field condition comprises changing at least one parameter of the period of time. In some embodiments, updating the magnetic field condition comprises changing at least one parameter of the motion path. In some embodiments, the method further comprises correlating the at least one event to at least one of (i) the time-varying force, (ii) the at least one period of time, or (iii) the at least one motion path. In some embodiments, the method further comprises selecting the magnetic field condition to have (i) a plurality of ranges of the timevarying force applied over (ii) a plurality of periods of time, (iii) a plurality of motion paths, or (iv) any combination of (i) - (iii). In some embodiments, the method further comprises: (a) applying the magnetic field having the magnetic field condition; (b) detecting a plurality of events caused by applying the magnetic field; and (c) aggregating the plurality of events to determine a statistical quality of data for each of the events, wherein each data for each of the events comprises a bond survival probability. In some embodiments, each data for each of the events has one or more types of statistical quality determined at least from a cardinality of each data. In some embodiments, the method further comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of ranges of the time-varying force at a predetermined time of the plurality of periods of time. In some embodiments, the method further comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of periods of time at a predetermined range or magnitude of the plurality of ranges of the time-varying force. In some embodiments, the method further comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of ranges of the time-varying force and (iii) the plurality of periods of time. In some embodiments, the method further comprises using the at least one motion path to generate the plurality of ranges of the time-varying force. In some embodiments, the method further comprises generating the plurality of periods of time by: (a) using the at least one motion path, wherein each period of time comprises one or more velocity profiles or one or more acceleration profiles of a movement of a magnetic source used to generate the magnetic field; or (b) using one or more excitation profiles of the magnetic source, wherein each excitation profile comprises one or more magnetic strength profiles or one or more magnetic strength duration profiles. In some embodiments, the method further comprises: (a) determining a bond survival probability between the capture moiety and the analyte based on the at least one event; (b) generating a surface plot of (i) the bond survival probability against (ii) the magnetic field condition; (c) extracting a characteristic time from the surface plot to determine (i) a diffusion relaxation time and (ii) dissociation constants; and (d) using the dissociation constants to identify the analyte as a lead candidate drug for a disease. In some embodiments, the bond survival probability comprises a dissociation chance of one or more molecular interactions between thecapture moiety and the analyte. In some embodiments, applying the magnetic field comprises acting on the magnetically-attractable particle using the time-varying force. In some embodiments, the time varying force comprises a peak force or a root mean square (rms) force of at least about 1 piconewton (pN), 10 pN, 100 pN, 1000 pN, or greater. In some embodiments, applying the magnetic field comprises acting on the magnetically-attractable particle using the at least one period of time. In some embodiments, the at least one period of time comprises at most about 1000 microseconds (ps), 100 ps, 10 ps, or less. In some embodiments, applying the magnetic field comprises acting on the magnetically-attractable particle using the at least one motion path relative to the magnetically-attractable particle. In some embodiments, the at least one motion path is (i) along a vertical axis of the magnetically-attractable particle, (ii) along a vertical axis spatially separated from the vertical axis in (i), (iii) along a horizontal plane parallel to the surface, or (iv) any combination of (i) - (iii). In some embodiments, the at least one motion path is a linear motion path or a nonlinear motion path comprising a path distance of about 0 mm to 100 mm. In some embodiments, the method further comprises applying the magnetic field to the magnetically- attractable particle by using an external or an integrated (i) magnetic source, (ii) electromagnetic source, or (iii) a combination of (i) and (ii). In some embodiments, the method further comprises applying a force to the force transmiter by using an external or an integrated (i) electrokinetic source, (ii) dielectrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v). In some embodiments, the capture moiety comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some embodiments, the analyte comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some embodiments, the force transmitter comprises at least one particle configured to couple to at least one analyte, and wherein the at least one particle comprises (i) a magnetic bead, (ii) a dielectric bead, (iii) a polystyrene bead, (iv) a charged molecule, or (v) any combination (i) - (iv). In some embodiments, the force transmitter comprises (i) a nucleic acid, (ii) a secondary antibody coupled to a nucleic acid, (iii) a secondary antibody coupled to a magnetic bead, (iv) a secondary antibody coupled to a dielectric bead, (v) a secondary antibody coupled to a polystyrene bead, (vi) a bead modified with at least one fluorescent molecule, or (vii) any combination (i) - (vi). In some embodiments, the method further comprises coupling the capture moiety to the surface using a covalent bond or a non-covalent bond. In some embodiments, the method further comprises directing the solution to at least one array portion of the surface, wherein each array portion comprises at least 1, 10, 100, or more individually programmable portions. In some embodiments, each individually programmableportion comprises at least 1, 10, 100, 1000, or more individual programmable assay areas. In some embodiments, each individually programmable assay area comprises a surface area of at least about 0.01 square millimeters (mm2), 0.1 mm2, 1.0 mm2, 10 mm2, 100 mm2, or greater In some embodiments, the method further comprises using at least one sensor configured to: (a) view the at least one event by an optical viewer; and (b) detect the at least one event by an optical sensor. In some embodiments, the least one sensor comprises (i) an optical sensor, (ii) an electromagnetic sensor, (iii) a mechanical sensor, (iv) a chemical sensor, or (v) any combination of (i) - (iv). In some embodiments, the method further comprises: (a) directing, by a flow cell, the solution having the analyte to a reservoir proximate to the surface; and (b) suspending, by a top surface operatively coupled to the reservoir, the solution in a volume of the reservoir. In some embodiments, the method further comprises coupling the capture moiety to the surface by flowing a gas phase or a liquid phase to deposit ((3 -Aminopropyl) trimethoxy silane) (ATPMS), ((3- Aminopropyl)triethoxysilane) (ATPES), or ((3-Glycidyloxypropyl) trimethoxysilane) (GOPS).

[0005] In another aspect, disclosed herein is a cartridge for identifying an analyte, the cartridge comprising: a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; a reservoir configured to suspend a solution proximate to the surface, wherein the solution comprises the analyte coupled to the magnetically-attractable particle; array portions of the surface configured to assay the analyte under application of a magnetic field, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and at least one sensor configured to detect at least one event of the analyte caused by the application of the magnetic field having the magnetic field condition, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii). In some embodiments, at least one event comprises a dissociation event between the capture moiety and the analyte. In some embodiments, the identifying comprises determining a bond survival probability between the capture moiety and the analyte based on the at least one event. In some embodiments, the bond survival probability comprises a dissociation chance of one or more molecular interactions between the capture moiety and the analyte. In some embodiments, the bond survival probability is used to identify the analyte as a lead candidate drug for a disease. In some embodiments, the magnetic field is configured to act on the magnetically-attractable particle with the time-varying force. In some embodiments, the time varying force comprises a peak force or a root mean square (rms) force of at least about 1 piconewton (pN), 10 pN, 100 pN, 1000 pN, or greater. In some embodiments, the magnetic field is configured to act on the magnetically-attractable particle overthe at least one period of time. In some embodiments, the at least one period of time comprises at most about 1000 ps (sec), 100 psec, 10 psec, or less. In some embodiments, the magnetic field is configured to act on the magnetically-attractable particle over the at one least motion path relative to the magnetically-attractable particle. In some embodiments, the at least one motion path is (i) along a vertical axis of the magnetically-attractable particle, (ii) along a vertical axis spatially separated from the vertical axis in (i), (iii) along a horizontal plane parallel to the surface, or (iv) any combination of (i) - (iii). In some embodiments, the at least one motion path is a linear motion path or a nonlinear motion path comprising a path distance of about 0 mm to 100 mm. In some embodiments, the cartridge is configured to apply the magnetic field to the magnetically-attractable particle by coupling to an external or an integrated (i) magnetic source, (ii) electromagnetic source, or (iii) a combination of (i) and (ii). In some embodiments, the cartridge is configured to apply a force to the force transmitter by coupling to an external or an integrated (i) electrokinetic source, (ii) dielectrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v). In some embodiments, the capture moiety comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some embodiments, the analyte comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some embodiments, the force transmitter comprises at least one particle configured to couple to at least one analyte, and wherein the at least one particle comprises (i) a magnetic bead, (ii) a dielectric bead, (iii) a polystyrene bead, (iv) a charged molecule, or (v) any combination (i) - (iv). In some embodiments, the force transmitter comprises (i) a nucleic acid, (ii) a secondary antibody coupled to a nucleic acid, (iii) a secondary antibody coupled to a magnetic bead, (iv) a secondary antibody coupled to a dielectric bead, (v) a secondary antibody coupled to a polystyrene bead, (vi) a bead modified with at least one fluorescent molecule, or (vii) any combination (i) - (vi). In some embodiments, the capture moiety is coupled to the surface using a covalent bond or a non-covalent bond. In some embodiments, at least one array portion of the surface comprises at least 1, 10, 100, or more individually programmable portions. In some embodiments, each individually programmable portion comprises at least 1, 10, 100, 1000, or more individual programmable assay areas. In some embodiments, each individually programmable assay area comprises a surface area of at least 0.01 square millimeters (mm2), 0.1 mm2, 1.0 mm2, 10 mm2, 100 mm2, or greater In some embodiments, the at least one sensor comprises: an optical viewer configured to optically view the at least one event; and an optical sensor configured to optically detect the at least one event. In some embodiments, the least one sensor comprises (i) an optical sensor, (ii) an electromagnetic sensor, (iii) a mechanical sensor, (iv)a chemical sensor, or (v) any combination of (i) - (iv). In some embodiments, the cartridge further comprises: a flow cell configured to direct the solution having the analyte to the reservoir proximate to the surface; and a top surface operatively coupled to the reservoir configured to suspend the solution in a volume of the reservoir; and In some embodiments, coupling the capture moiety to the surface comprises flowing a gas phase or a liquid phase to deposit ((3 -Aminopropyl) trimethoxysilane) (ATPMS), ((3-Aminopropyl)triethoxysilane) (ATPES), or ((3- Glycidyloxypropyl) trimethoxysilane) (GOPS). In some embodiments, the cartridge is configured to couple to an instrument for identifying the analyte.

[0006] In another aspect, disclosed herein is a system comprising at least one processor and instructions executable by the at least one processor to cause the at least one processor to perform operations comprising: (a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; (b) using the capture moiety to capture the analyte; (c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and (d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

[0007] In another aspect, disclosed herein is a computer-implemented method for identifying an analyte, the method comprising: (a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; (b) using the capture moiety to capture the analyte; (c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and (d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

[0008] Additional aspects and advantages of the present disclosure will become readily apparent from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obviousrespects, all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature and not as restrictive.INCORPORATION BY REFERENCE

[0009] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the present disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The novel features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the present disclosure are utilized, and the accompanying drawings of which:

[0011] FIG. 1 depicts an example system configured to perform methods herein, in accordance with some embodiments.

[0012] FIGs. 2A-2B depict example energy landscapes of molecular interaction, in accordance with some embodiments. FIG. 2A depicts an example energy landscape with a single energy barrier. FIG. 2B depicts an example energy landscape with a cascade of energy barriers.

[0013] FIGs. 3A-3F depict an example cartridge configured to perform methods herein, in accordance with some embodiments. FIG. 3A depicts an example cartridge, which can include a reservoir, an assay area, and a substrate. FIG. 3B depicts an example cartridge, which can further include a calibration layer. FIG. 3C depicts an example cartridge, which can include an assay area having individually programmable assay arrays. FIGs. 3D-3E depict an example cartridge, which the assay area thereof can be functionalized or activated using methods herein. FIG. 3F depicts a characterization graph of functionalization or activation generated by X-ray photoelectron spectroscopy.

[0014] FIGs. 4A-4C depict example plots generated by methods herein for determining bond survival probability, in accordance with some embodiments. FIG. 4A depicts an example surface plot of bond survival probability for different ranges of applied force and time. FIG. 4B depicts an example line plot of bond survival probability for different strengths of applied force and rangesof time. FIG. 4C depicts an example plot of characteristic times for different ranges of applied force.

[0015] FIGs. 5A-5G depict example methods of using a force source to apply an external force to a molecular complex, in accordance with some embodiments. FIG. 5A depicts an example of a force source positioned relative to a plurality of molecular complexes. FIG. 5B depicts an example plot of determining the applied force by position of the force source relative to a molecular complex. FIGs. 5C-5E depict example methods of generating a range of applied forces by positioning the force source relative to the plurality of molecular complexes. FIGs. 5F-5G depict example methods of generating a range of applied forces by moving the force source along a motion path relative to the plurality of molecular complexes.

[0016] FIGs. 6A-6B depict example plots of types of applied force, in accordance with some embodiments. FIG. 6A depicts an example of a continuous applied force, which can be used to increase or decrease the rate of the applied force over time. FIG. 6B depicts an example of a discrete applied force, which can be used to apply the force in a step or piece-wide manner over time.

[0017] FIG. 7 depicts an example computing device configured to perform methods herein, in accordance with some embodiments.

[0018] FIG. 8 depicts an example web or mobile application provision system configured to perform methods herein, in accordance with some embodiments.

[0019] FIG. 9 depicts an example cloud-based web or mobile application provision system configured to perform methods herein, in accordance with some embodiments.DETAILED DESCRIPTION

[0020] While various embodiments of the present disclosure have been shown and described herein, such embodiments are provided by way of example only. Numerous variations, changes, or substitutions may occur without departing from the present disclosure. It should be understood that various alternatives to the embodiments of the present disclosure described herein may be employed.Overview

[0021] Recognized herein is a need for systems and methods that can determine molecular affinity using high-throughput methods that are easier and quicker to use and yet can provide higher- resolution data compared to other methods. Systems and methods herein can be applied to drug discovery and development for improving healthcare outcomes. In some cases, systems andmethods herein can be configured to use a type of dynamic force spectroscopy (DFS) for determining molecular affinity.

[0022] As a background, biological instructions can be executed by interactions at the molecular level. For example, a typical underlying interaction can be governed by bonds of different types that form by participating molecules. In some cases, bonds can be weak bonds that can include non-covalent bonds, e.g., hydrogen bonds, coulombic bonds, hydrophobic interactions, or van der Waals interactions. In some cases, bonds can be strong bonds, e.g., covalent bonds. A practical understanding of biological processes may require a better understanding of these bonds as they occur in biology.

[0023] Further, formation and dissociation of molecular bonds, e.g., non-covalent bonds, can be non-deterministic due to their interaction with a thermal reservoir. Such interactions have been theoretically studied by Kramers. (See Kramers, “Brownian motion in a field of force and the diffusion model of chemical reactions.” Physica 7.4 (1940): 284-304, which is incorporated by reference herein in its entirety). Also, Evans theorized that, to understand molecular interactions and bond disruptions thereof, scientists may need to consider not only the applied force utilized by some methods to disrupt or detach bonds but also the rate of loading of the force that is applied to the molecular interactions over time. (See Evans, “Probing the relation between force - lifetime - and chemistry in single molecular bonds.” Annual review of biophysics and biomolecular structure 30.1 (2001): 105-128, which is incorporated by reference herein in its entirety).

[0024] Additionally, some scientists have theorized that biological processes may be determined by a spectrum of molecular bonds rather than merely formation and disruption of a single molecular bond. Thus, to better understand a type of molecular interaction, systems and methods herein can assay multiple instances of molecular interactions by interrogating the interactions over different ranges of forces, different rates force loading, and different timescales. Compared to other methods, systems and methods herein can obtain more relevant and improved multi-faceted data to determine and characterize molecular bonds conveniently and rapidly.

[0025] For example, compared to the present disclosure, surface plasmon resonance (SPR) techniques can be deficient when applied to determining drug efficacy or drug toxicity during drug discovery, e.g., efficacy or toxicity of a biologic. First, methods that utilize SPR techniques can be cumbersome and time-consuming because they may require analyzing many exhaustive series of analyte concentrations to determine the desired outputs for molecular affinity, e.g., association constants (kon) and dissociation constants (koff). Second, because methods that utilize SPR techniques may merely rely on near equilibrium dissociation, they can require monitoring for a long time, e.g., hours or days for tight binding biologies. Third, due to a low signal-to-noise ratio(SNR), methods that utilize SPR techniques may not be able to accurately measure such small signals associated with dissociation without undesirably increasing detection or measurement errors.

[0026] As an example of these deficiencies, methods that utilize SPR-based extraction of konand koff may require many, exhaustive experimental runs, each run varying analyte concentration, which can be time-consuming, costly, and prone to error. The association rate (Ron), one of the direct outputs of SPR techniques, can be a function of kon, koff, and analyte concentration. So, SPR techniques typically require varying analyte concentration numerous times to obtain kon. Such approaches to accurately determine konand koff can be very time-consuming and unacceptably error prone thereby increasing costs for determining quality drug candidates during drug discovery. The time-consuming nature of such approaches can result from typical monoclonal antibody dissociation constants that can result in nearly imperceptible dissociation behavior, e.g., koff can typically be on the order of 10'5s'1to 10'4s'1. Such long experimental assays can reduce the accuracy of extracted parameters such as konand koff.

[0027] As another example of these deficiencies illustrated in FIGs. 2A-2B, surface plasmon resonance (SPR) techniques can be deficient because they may study molecular affinity at equilibrium or near equilibrium so such techniques can only investigate observable events at or near equilibrium. For example, as illustrated in FIG. 2A, consider a molecular interaction landscape in energy space. Energy 601 can be considered in relation to molecular position 602. In some cases, the behavior of the interaction may be dominated by a single activation barrier 605. For example, energy landscape at equilibrium 603 and under force 604 may both be dominated by activation barrier 605. However, in practice, molecular bonds can involve widely distributed types of intermolecular forces ranging from, e.g., hydrogen bonds, Van der Waals forces, forces arising from solvation and Coulombic forces, and combinations of each type. Each type of intermolecular force can contribute to the energy landscape. For example, as illustrated in FIG. 2B, different types of intermolecular forces can result in multiple activation barriers 615 and 616. In some cases, the equilibrium behavior 613 can be dominated by activation barrier 616, which may mask the other activation barrier 615. However, under an applied force associated with 614, methods herein can unmask the dominance of activation barrier 616 to reveal or observe the effects of activation barrier 615. Thus, methods herein can use dynamic force spectroscopy (DFS) to provide a technical solution for better understanding the underlying behavior of molecular interactions. Such understanding can be applied to practically improving the use of molecular affinity in drug discovery or development.

[0028] In contrast, the present disclosure utilizing DFS, can improve over SPR techniques and other methods by performing a single assay to characterize the full scope of molecular interactions thereby determining molecular affinity. For example, systems and methods herein can perform a single assay that can improve upon SPR techniques by obtaining more data in less time for desired outputs, konand koff. Alternatively, systems and methods herein can perform a single assay that obtains less data that is at least as accurate for desired outputs, konand koff. In some cases, the assay can be a multiplexed assay. Also, systems and methods herein can uses dynamic force spectroscopy (DFS) to detect molecular bond behavior under different conditions of applied force, duration of the applied force, and rate of loading of the applied force. Using different conditions can be useful for making multidimensional observations described herein.Systems and methods

[0029] FIG. 1 depicts a high-level architecture of system 100, which can be configured to perform methods herein, e.g., identify analyte 270. System 100 can include cartridge 200, force source 300, sensor 400, and controller 500. In some embodiments, the cartridge 200 is configured to couple to an instrument for identifying analyte 270. In some cases, the terms “system” and “instrument” can be used interchangeably. In some cases, controller 500 can be configured as a computing system described herein to operate system 100.

[0030] FIGs. 3A-3C depict cartridge 200, which can be configured for identifying analyte 270 using methods herein. Cartridge 200 can be configured with reservoir 210, assay area 220, assay array 225, and substrate 230. In some cases, cartridge 200 can be a consumable or disposable cartridge. In some cases, the terms “cartridge” and “chip” can be used interchangeably.

[0031] FIG. 5A depicts analyte 270, which can be formed as molecular complex 800. Molecular complex 800 can include probe 260, analyte 270, and force transmitter 280. In some embodiments, the analyte 270 comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some embodiments, the capture moiety 260 comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some cases, the terms “probe” and “capture moiety” can be used interchangeably.Cartridges

[0032] System 100 can include cartridge 200 configured to implement methods herein. FIG. 3A depicts cartridge 200, which can be configured with substrate 230 for identifying analyte 270. In some embodiments, the cartridge 200 comprises a surface having a capture moiety 260 coupled thereto. In some cases, substrate 230 can include the surface having capture moiety 260 coupled thereto. In some cases, the surface can include a surface of assay area 220 or assay array 225. Insome embodiments, the method comprises directing a solution having the analyte 270 to a surface having a capture moiety 260 coupled thereto. In some embodiments, the method comprises using the capture moiety 260 to capture the analyte 270.

[0033] In some cases, substrate 230 can be functionalization or activated as described herein. In some cases, substrate 230 can be sized according to the assay being performed. In some cases, substrate 230 can be sized with a surface area, a width, or a length. In some cases, the width and length can be sized to obtain a surface area of substrate 230. In some cases, the surface area of substrate 230 can be at least about 1.0 square centimeters (cm2) 10 cm2, 100 cm2, and increments thereof, or greater. In some cases, the surface area of substrate 230 can be at most about 100 cm2, 10 cm2, 1.0 cm2, and increments thereof, or less. In some cases, substrate 230 can be fabricated from a suitable material for fluid handling, e.g., glass, quartz, silicon, and the like. In some cases, substrate 230 can be fabricated from a suitable material for biocompatibility, e.g., glass, quartz, silicon, and the like.

[0034] FIG. 3B depicts cartridge 200, which can be configured with calibration layer 240. Calibration layer 240 can be used to calibrate a position of some or all components of system 100, e.g., force source 300, with respect to a surface of assay area 220, assay array 225, or substrate 230. In some cases, determining or measuring the position during calibration can be nondestructive for any component of cartridge 200. In some cases, using prior calibration of force source 300, a force applied by force source 300 can be automatically determined, e.g., selfcalibrated. In some cases, calibration layer 240 can be fabricated by depositing a suitable conductive material on a surface of assay area 220, assay array 225, and substrate 230, e.g., a deposition of aluminum, copper, polysilicon, silicide, titanium, platinum, gold, and the like. In some cases, calibration layer 240 can be deposited with a surface area that covers a surface of substrate 230. In some cases, calibration layer 240 can be deposited with a surface area that covers a portion of the surface of substrate 230. For example, calibration layer 240 can be deposited with a surface area that convers a surface of assay area 220 or assay array 225.

[0035] In some cases, calibration enabled by calibration layer 240 can be a Z-axis or height calibration, e.g., a vertical axis along force source 300 and calibration layer 240. In some cases, calibration can be performed by electrical impedance based methods. For example, calibration layer 240 can be a conductive layer described herein. For example, force source 300 can be configured with a magnetic tip having a conductive material. During calibration, as force source 300 approaches calibration layer 240, the electrical impedance between the two can be measured or monitored to accurately determine a contact or near-contact position between force source 300 and calibration layer 240. In some cases, electrical impedance can be measured by measuringchanges in capacitance. In some cases, electrical impedance can be measured by measuring changes in resistance.

[0036] In some cases, calibration enabled by calibration layer 240 can be an x-y calibration, e.g., an x-y position of force source 300 along a plane relative to a surface of assay area 220, assay array 225, or substrate 230. For example, x-y calibration can be performed using image analysis from sensor 400. In some cases, image analysis can be performed by analyzing images of cartridge 200 obtained by sensor 400. For example, assay area 220, assay array 225, or substrate 230 can include geometric markings fabricated therewith from which x-y calibration can be performed.

[0037] FIGs. 3A-3B depict cartridge 200, which can be configured with reservoir 210. In some embodiments, the cartridge 200 comprises a reservoir 210 configured to suspend a solution proximate to the surface. In some cases, the surface can be a surface of assay area 220, assay array 225, or substrate 230, or any combination thereof. For example, assays herein can be performed in an aqueous environment (e.g., a fluid solution) contained by reservoir 210. In some embodiments, the solution comprises the analyte 270 coupled to the magnetically-attractable particle. In some cases, reservoir 210 can be configured as a flow cell. In some embodiments, the cartridge 200 comprises a flow cell configured to direct the solution having the analyte 270 to the reservoir 210 proximate to the surface. In some embodiments, the cartridge 200 comprises a top surface operatively coupled to the reservoir 210 configured to suspend the solution in a volume of the reservoir 210. In some embodiments, the method comprises directing, by a flow cell, the solution having the analyte 270 to a reservoir proximate 210 to the surface. In some embodiments, the method comprises suspending, by a top surface operatively coupled to the reservoir 210, the solution in a volume of the reservoir 210.

[0038] FIGs. 3A-3B depict reservoir 210, which can be sized according to the assay being performed for identifying analyte 270. In some cases, reservoir 210 can be sized with a volume, a width, a length, or a depth. In some cases, the width, length, and depth can be sized to obtain a volume of reservoir 210. In some cases, the volume of reservoir 210 can be at least about 0.1 milliliters (mL), 1.0 mL, 5 mL, and increments thereof, or greater. In some cases, the volume of reservoir 210 can be at most about 5 mL, 1.0 mL, 0.1 mL, and increments thereof, or less.

[0039] FIG. 3C depicts cartridge 200, which can be configured with assay area 220 for identifying analyte 270. Each assay area 220 can be individually programmable by selectively functionalizing or activating each of assay area 220 as described herein. Selectively functionalizing or activating each of assay area 220 can be used to perform multiplexed assays of analyte 270. For example, substrate 230 can include assay area 220 (or array portion). In some embodiments, at least one array portion 220 of the surface 230 comprises at least 1, 10, 100, or more individuallyprogrammable portions. In some embodiments, each array portion 220 comprises at least 1, 10, 100, or more individually programmable portions. In some cases, each array portion 220 comprises at most about 100, 10, or less individually programmable portions. In some embodiments, each individually programmable assay area 220 comprises a surface area of at least about 0.01 square millimeters (mm2), 0.1 mm2, 1.0 mm2, 10 mm2, 100 mm2, or greater. In some cases, each individually programmable assay array 220 comprises a surface area of at most about 100 square millimeters (mm2), 10 mm2, 1.0 mm2, 0.1 mm2, 0.001 mm2, or less. In some embodiments, the method comprises directing the solution to at least one array portion 220 of the surface 230.

[0040] FIG. 3C depicts assay area 220, which can be configured with assay array 225 for identifying analyte 270. Each assay array 225 can be individually programmable by selectively functionalizing or activating each of assay area 225 as described herein. Selectively functionalizing or activating each of assay array 225 can be used to perform multiplexed assays of analyte 270. For example, substrate 230 can include assay array 225. In some embodiments, each individually programmable portion 225 comprises at least 1, 10, 100, 1000, or more individual programmable assay areas. In some cases, each individually programmable portion 225 comprises at most 1000, 100, 10, or less individual programmable assay areas. In some cases, each individually programmable assay array 225 comprises a surface area of at least about 0.001 square millimeters (mm2), 0.01 mm2, 0.1 mm2, 1.0 mm2, 10 mm2, and increments therein, or greater. In some cases, each individually programmable assay array 225 comprises a surface area of at most about 10 square millimeters (mm2), 1.0 mm2, 0.1 mm2, 0.01 mm2, 0.001 mm2, and increments therein, or less. In some cases, the method comprises directing the solution to at least one assay array 225 of the surface 230.

[0041] In some cases, each of assay array 225 can be partitioned into a number of elements. For example, assay array 225 can be partitioned into a x by y array of elements. In some cases, x of the array of elements can include at least 1, 10, 1000, 10000, and increments thereof, or more individually programmable elements. In some cases, x of the array of elements can include at most 10000, 1000, 100, 10, and increments thereof, or less individually programmable elements. In some cases, y of the array of elements can include at least 1, 10, 1000, 10000, and increments thereof, or more individually programmable elements. In some cases, y of the array of elements can include at most 10000, 1000, 100, 10, and increments thereof, or less individually programmable elements. In some cases, each element of each assay array 225 can be individually programmable by selectively functionalizing or activating each element of each assay array 225 as described herein.

[0042] In some embodiments, the method comprises coupling the capture moiety 260 to the surface 230 using a covalent bond or a non-covalent bond. In some cases, cartridge 200 can includeassay area 220, assay array 225, and substrate 230, any of which can include a surface functionalized or activated as described herein. For example, any or all of assay area 220, assay array 225, or substrate 230 can be chemically modified to strongly attach or immobilize molecules of interest using molecular bonds, e.g., covalent bonds. As illustrated in FIG. 3D, assay area 220, assay array 225, or substrate 230 can be chemically modified using epoxy silane 250. In some cases, chemical modification can be performed using amino silane. In some cases, chemical modification can be performed by flowing a gas or liquid phase of epoxy silane 250 or amino silane to contact a surface of assay area 220, assay array 225, or substrate 230. In some cases, a structure of epoxy silane 250 or amino silane can include ((3 -Aminopropyl) trimethoxysilane) (ATPMS), ((3 -Aminopropyl)tri ethoxy silane) (ATPES), or ((3-Glycidyloxypropyl) trimethoxy silane) (GOPS). In some embodiments, the method comprises coupling the capture moiety 260 to the surface 230 by flowing a gas phase or a liquid phase to deposit ((3 -Aminopropyl) trimethoxy silane) (ATPMS), ((3 -Aminopropyl)tri ethoxy silane) (ATPES), or ((3-Glycidyloxypropyl) trimethoxysilane) (GOPS).

[0043] In some cases, as illustrated in FIG. 3E, a gas phase or a liquid phase of epoxy silane 250 can deposit a self-assembling monolayer of epoxy groups 265 to functionalize or activate assay area 220, assay array 225, or substrate 230. For example, epoxy groups 265 can react with probe 260. In some embodiments, the capture moiety 260 comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii). In some cases, epoxy groups 265 may form a strong covalent bond with probe 260. FIG. 3F depicts an outcome of functionalization or activation of assay area 220, assay array 225, or substrate 230 using via X-ray photoelectron spectroscopy.

[0044] In some embodiments, the capture moiety 260 is coupled to the surface using a covalent bond or a non-covalent bond. In some cases, the surface is a surface of assay area 220, assay array 225, or substrate 230. For example, a strong covalent bond between assay area 220, assay array 225, or substrate 230 and probe 260 can be useful for interrogating and determining a molecular interaction, e.g., molecular affinity, between analyte 270 and probe 260. In some cases, analyte 270 can include a corresponding antigen for an antibody of probe 260. For example, probe 260 can be sufficiently immobilized to avoid detaching from assay area 220, assay array 225, or substrate 230 under a force applied by force source 300. Such immobilization can be useful because the molecular affinity of interest is between analyte 270 and probe 260. The force applied by force source 300 can be sufficient to decouple analyte 270 from probe 260 but leave probe 260 attached to assay area 220, assay array 225, or substrate 230.Force sources and transmitters

[0045] System 100 can include force source 300 and force transmitter 280 configured to implement methods herein. In some embodiments, the analyte 270 is coupled to a force transmitter 280 comprising a magnetically-attractable particle. In some cases, the terms “force transmitter” and “magnetically-attractable particle” can be used interchangeably. In some cases, analyte 270 can be coupled to, e.g., chemically attached, force transmitter 280. Force source 300 can be applied to force transmitter 280 to cause a force to act on or actuate force transmitter 280 and thereby transmit the applied force to analyte 270. In some cases, the applied force can be sufficient to disrupt or detach bonds between analyte 270 and probe 260 but insufficient to detach probe 260 from assay area 220, assay array 225, or substrate 230. For example, the applied force can disrupt or detach bonds by pulling force transmitter 280 and analyte 270 coupled thereto away from probe 260. In some embodiments, the force transmitter 280 comprises at least one particle configured to couple to at least one analyte 270. In some embodiments, the analyte 270 is coupled to a force transmitter 280 comprising a magnetically-attractable particle. In some cases, force transmitter 280 may not be limited to a magnetically-attractable particle. For example, force transmitter 280 can include any particle susceptible to a force applied by force source 300. In some embodiments, the at least one particle comprises (i) a magnetic bead, (ii) a dielectric bead, (iii) a polystyrene bead, (iv) a charged molecule, or (v) any combination (i) - (iv). In some cases, magnetic beads can include superparamagnetic beads such as Dynabeads® beads or Sera-Mag™ beads.

[0046] In some embodiments, the cartridge 200 is configured to apply the magnetic field to the magnetically-attractable particle by coupling to an external or an integrated (i) magnetic source, (ii) electromagnetic source, or (iii) a combination of (i) and (ii). In some cases, force source 300 may not be limited to (i) or (ii). For example, force source 300 can include any source that is configured to act on or actuate force transmitter 280. In some embodiments, cartridge 200 is configured to apply a force to the force transmitter 280 by coupling to an external or an integrated (i) electrokinetic source, (ii) di electrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v). In some embodiments, the method comprises applying a force to the force transmitter 280 by using an external or an integrated (i) electrokinetic source, (ii) di electrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v).Use of force sources and transmitters

[0047] FIGs. 5A-5G illustrate methods herein for identifying or characterizing analyte 270. In some cases, methods herein may perform dynamic force spectroscopy (DFS) using force source 300. In some cases, force source 300 can be configured as an electromagnetic (EM) source. Forexample, as illustrated in FIG. 5A, consider a cross-sectional profile view of system 100, which can include cartridge 200 and force source 300. Cartridge 200 can include substrate 230 with functionalized or activated assay area 220 or assay array 225. Assay area 220 or assay array 225 can be functionalized or activated to immobilize a plurality of molecular complexes 800. Molecular complex 800 can include probe 260 (e.g., an antibody), analyte 270 (e.g., an antigen), and force transmitter 280, each described herein. In some cases, force transmitter 280 can be a magnetic bead susceptible to application of force by the EM source. In some cases, force source 300, e.g., the EM source, can be configured to apply a magnetic field condition describe herein to force transmitter 280. The magnetic field condition can include different strengths of applied force by varying a distance of the EM source relative to molecular complexes 800.

[0048] For example, molecular complex 1, being closer to the EM source than complexes 2 and 3, may be susceptible to a stronger applied force than complexes 2 and 3. Molecular complex 2, being closer to the EM source than molecular complex 3, may be susceptible to a stronger applied force than molecular complex 3. In some cases, molecular complex 800 (e.g., complex 1) that is positioned purely along a vertical axis of EM source may be susceptible to a different applied force than molecular complex 800 (e.g., complex 2 and 3) that is offset from the vertical axis of the EM source. For example, molecular complexes 2 and 3 may be susceptible to a different tangential applied force (e.g., along an axis normal to the vertical axis of the EM source.

[0049] For example, FIG. 5B illustrates spatial variation of the applied force. The x-axis of FIG. 5B is the tangential or lateral distance away from the vertical axis of the EM source in units of micrometers (pm); the y-axis is the estimated force applied (e.g., in nanonewtons, nN) by the EM source to force transmitter 280, e.g., a 5.5 pm diameter magnetic bead. Each series 811.1 and 811.2 can represent, for example, the case where the EM source is about 10 pm above the surface (811.1) of substrate 230 and 20 pm above the surface (811.2) of substrate 230. Characterizing the applied force along the lateral distance from the EM source can be useful for simultaneously applying a range of forces across assay area 220 thereby simplifying the process of generating data by methods herein.

[0050] FIGs. 5C-5G illustrate the behavior of molecular complexes 800 as force source 300 is applied at different positions (FIGs. 5C-5E) or along different motion paths (FIGs. 5F-5G) relative to molecular complexes 800. Applying force source 300 in either manner can be useful for assaying molecular interactions across assay area 220 or assay array 225. For example, FIGs. 5C- 5E illustrate the behavior of molecular complexes 800 as force source 300 is applied at different positions along a vertical axis relative to molecular complexes 800. In some cases, force source 300 can be configured to move along a range of positions along the vertical axis. In some cases,force source 300 can move about 0 millimeters (mm) to 10 mm, 10 mm - 20 mm, 20 mm - 30 mm, and increments therein, or greater.

[0051] For example, FIG. 5C illustrates force source 300 positioned far from molecular complexes 800 such that the applied force may not be sufficient for bond disruption, e.g., analyte 270 may not detach from probe 260. FIG. 5D illustrates that, as force source 300 is positioned closer to molecular complexes 800, the increasing force applied to molecular complexes 800 will be sufficient for some bond disruption, e.g., some of analyte 270 may detach from probe 260. In some cases, bond disruption may occur near the vertical axis of force source 300 to create an area devoid 831 of molecular complexes 800. FIG. 5E illustrates that, as force source 300 is positioned in contact (or in near contact) with molecular complexes 800, the increasing force applied to molecular complexes 800 will be sufficient for even more bond disruption, e.g., more of analyte 270 may detach from probe 260. In some cases, force source 300 can create a larger area devoid 841 of molecular complex 800.

[0052] In some cases, an excitation of force source 300 can be kept low but use a closer proximity to molecular complexes 800 to still result in an applied force sufficient to cause bond disruption of molecular complexes 800. In some cases, an excitation of force source 300 can be high but use a further proximity from molecular complexes 800 to still result in an applied force sufficient to cause bond disruption of molecular complexes 800.

[0053] In some embodiments, the method comprises generating the plurality of periods of time by using the at least one motion path. In some embodiments, each period of time comprises one or more velocity profiles or one or more acceleration profiles of a movement of a magnetic source 300 used to generate the magnetic field. In some cases, force source 300 can be configured to move along a range of positions of an x-y plane relative to assay area 220, assay array 225, or substrate 230. In some cases, force source 300 can move along the x axis of the x-y plane of about 0 millimeters (mm) to 50 mm, 50 mm - 100 mm, 100 mm - 150 mm, and increments therein, or greater. In some cases, force source 300 can move along the y axis of the x-y plane of about 0 millimeters (mm) to 50 mm, 50 mm - 100 mm, 100 mm - 150 mm, and increments therein, or greater.

[0054] For example, FIGs. 5F-5G illustrate the behavior of molecular complexes 800 as force source 300 is applied along different motion paths 851 (e.g., spatial movement) relative to molecular complexes 800. In some cases, motion paths 851 are along a surface of a plane (e.g., x- y plane) relative to assay array 225. FIG. 5F illustrates molecular complexes 800 immobilized to assay array 225 but without application of force source 300. In some cases, molecular complexes 800 may be randomly distributed or immobilized across assay array 225. In some cases, molecularcomplexes 800 may be distributed or immobilized across assay array 225 at predetermined positions. FIG. 5G illustrates the behavior of molecular complexes 800 as force source 300 is applied along different motion paths 851 relative to molecular complexes 800. For example, as force source 300 is moved along motion path 851 relative molecular complexes 800, the force applied to molecular complexes 800 along motion path 851 may be sufficient for some bond disruption, e.g., some of analyte 270 may detach from probe 260.

[0055] In some cases, a combination of positions and motion paths of force source 300 can be used simultaneously to identify or characterize analyte 270. In some cases, an excitation of force source 300 in combination with positions and motion paths can be used to identify or characterize analyte 270. Such combinations can be useful for generating higher-resolution characteristics of molecular interactions in a single assay.

[0056] In some cases, force source 300 can be configured with a condition. In some cases, the condition can cause a force to act on or actuate force transmitter 280. In some cases, the condition can be a magnetic field condition that causes a magnetic force to act on or actuate force transmitter 280. In some cases, the magnetic force acting on force transmitter 280 can be sufficient to cause disruption or detachment of analyte 270 from probe 260. In some embodiments, the magnetic field has a magnetic field condition that is sufficient to direct the analyte 270 and the magnetically- attractable particle coupled thereto away from the capture moiety 260.

[0057] In some embodiments, the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii). In some embodiments, the method comprises applying a magnetic field to the magnetically- attractable particle. In some cases, the force applied by force source 300 can be changed by adjusting a position of or moving force source 300 relative to force transmitter 280.

[0058] In some embodiments, the magnetic field is configured to act on the magnetically- attractable particle with the time-varying force. In some embodiments, applying the magnetic field comprises acting on the magnetically-attractable particle using the time-varying force. For example, force source 300 can cause a force to act on or actuate force transmitter 280 and analyte 270 coupled thereto. The force may be sufficient to cause the event, e.g., disruption or detachment between analyte 270 and probe 260. In some embodiments, the time varying force comprises a peak force or a root mean square (rms) force of at least about 1 piconewton (pN), 10 pN, 100 pN, 1000 pN, or greater. In some cases, the time varying force comprises a peak force or a root mean square (rms) force of at most about 1000 pN, 100 pN, 10 pN, 1 pN, or less. In some cases, the peak force or the root mean square (rms) force can include increments therein of 1 pN, 10 pN, 100 pN, 1000 pN. In some embodiments, the method comprises using the magnetic field condition in toidentify at least one event of the analyte 270. In some embodiments, the method comprises correlating the at least one event to time-varying force. For example, controller 500 in combination with sensor 400 can record parameters of force source 300, e.g., state (position such as x), movement (velocity such as dx / dt), and amount of excitation (such as electrical current). Based on the parameters, controller 500, for any given output of the sensor 400 (e.g., microscope image) can determine applied force, duration of the applied force, and loading of the applied force.

[0059] In some embodiments, the magnetic field is configured to act on the magnetically- attractable particle 280 over the at least one period of time. In some embodiments, applying the magnetic field comprises acting on the magnetically-attractable 280 particle using the at least one period of time. For example, force source 300 can cause a force to act over a period of time (e.g., duration of force) on force transmitter 280 and analyte 270 coupled thereto. The duration of the force may be sufficient to cause the event, e.g., disruption or detachment between analyte 270 and probe 260. In some embodiments, the at least one period of time comprises at most about 1000 microseconds (psec), 100 psec, 10 psec, 1 psec, or less. In some cases, the at least one period of time comprises at least about 1 ps, 10 psec, 100 psec, 1000 psec, or more. In some cases, the period of time can include increments therein of 1 ps sec, 10 psec, 100 psec, 1000 psec. In some embodiments, the method comprises using the magnetic field condition to identify at least one event of the analyte 270. In some embodiments, the method comprises correlating the at least one event to at least one period of time. For example, controller 500 in combination with sensor 400 can record parameters of force source 300, e.g., state (position such as x), movement (velocity such as dx / dt), and amount of excitation (such as electrical current). Based on the parameters, controller 500, for any given output of the sensor 400 (e.g., microscope image) can determine applied force, duration of the applied force, and loading of the applied force.

[0060] In some embodiments, the magnetic field is configured to act on the magnetically- attractable particle 280 over the at one least motion path relative to the magnetically-attractable particle. In some embodiments, applying the magnetic field comprises acting on the magnetically- attractable particle using the at least one motion path relative to the magnetically-attractable particle. In some embodiments, the method comprises using the at least one motion path to generate the plurality of ranges of the time-varying force. For example, as illustrated in FIGs. 5F-5G, force source 300 can cause a force to act over a motion path relative to force transmitter 280 and analyte 270 coupled thereto. The motion path of the force may be sufficient to cause the event, e.g., disruption or detachment between analyte 270 and probe 260. In some embodiments, the at least one motion path is (i) along a vertical axis of the magnetically-attractable particle, (ii) along a vertical axis spatially separated from the vertical axis in (i), (iii) along a horizontal plane parallelto the surface, or (iv) any combination of (i) - (iii). In some cases, the surface can be a surface of substrate 230, assay area 220, assay array 225, or any combination thereof. In some embodiments, the at least one motion path is a linear motion path or a nonlinear motion path comprising a path distance of about 0 mm to 100 mm. In some cases, the path distance can be at least about 1 mm, 10 mm, 100 mm, and increments therein, or greater. In some cases, the path distance can be at most about 100 mm, 10 mm, 1 mm, and increments therein, or less. In some embodiments, the method comprises using the magnetic field condition in to identify at least one event of the analyte 270. In some embodiments, the method comprises correlating the at least one event to at least the at least one motion path. For example, controller 500 in combination with sensor 400 can record parameters of force source 300, e.g., state (position such as x), movement (velocity such as dx / dt), and amount of excitation (such as electrical current). Based on the parameters, controller 500, for any given output of the sensor 400 (e.g., microscope image) can determine applied force, duration of the applied force, and loading of the applied force.

[0061] In some cases, force source 300 can be configured with a means of excitation. The excitation of force source 300 can allow force source 300 to operate as an adjustable force source to change, e.g., increase or decrease, the force applied to force transmitter 280. In some cases, an excitation of force source 300 can be configured to generate (i) the time-varying force or (ii) the at least one period of time. For example, force source 300 can be configured as a magnetic source or an electromagnetic (EM) source excited by an electrical current or voltage. In some cases, the method comprises generating the plurality of time-varying forces by using one or more excitation profiles of the magnetic source. In some embodiments, the method comprises generating the plurality of periods of time by using one or more excitation profiles of the magnetic source. In some embodiments, each excitation profile comprises one or more magnetic strength profiles or one or more magnetic strength duration profiles.

[0062] In some cases, force source 300 can be configured to use force loading to identify analyte 270. For example, FIGs. 6A-6B illustrate different types of force loading that can be used by methods herein. FIG. 6A illustrates how the applied force can vary in time in a continuous manner. For example, the rate of applied force versus time can be continuously decreased over time 901 or continuously increased over time 902. FIG. 6B illustrates how the applied force can vary in time in a discrete manner. For example, the force can be applied in a step or piece-wise manner as depicted by 910-915. Such discrete application of force can be useful for assaying molecular interactions on a pre-determined timescale.

[0063] In some embodiments, the method comprises selecting the magnetic field condition to have (i) a plurality of ranges of the time-varying force applied over (ii) a plurality of periods of time,(iii) a plurality of motion paths, or (iv) any combination of (i) - (iii). In some embodiments, the method comprises applying the magnetic field having the magnetic field condition. In some embodiments, the method comprises detecting a plurality of events caused by applying the magnetic field. For example, the plurality of events can be more than one bond disruption between analyte 270 and probe 260. For example, the plurality of events can be more than one detachment between more than probe 260 and more than one analyte 270.Sensors

[0064] System 100 can include sensor 400 configured to implement methods herein. For example, sensor 400 can be configured to detect or determine disruption or detachment of bonds between analyte 270 and probe 260, e.g., between the antibody and the antigen. In some embodiments, the cartridge 200 comprises at least one sensor 400 configured to detect at least one event of the analyte 270 caused by the application of the magnetic field having the magnetic field condition. In some embodiments, at least one event comprises a dissociation event between the capture moiety 260 and the analyte 270. For example, the dissociation event can include a bond disruption between capture moiety 260 and analyte 270. For example, the dissociation event can include a detachment of capture moiety 260 and analyte 270.

[0065] In some embodiments, the least one sensor 400 comprises (i) an optical sensor, (ii) an electromagnetic sensor, (iii) a mechanical sensor, (iv) a chemical sensor, or (v) any combination of (i) - (iv). Sensor 400 may not be limited to (i) - (iv). Sensor 400 can include any combination of sensors configured to detect detachment of analyte 270 from probe 260. For example, detachment of analyte 270 from probe 260 can be optically detected by a microscope in combination with an optical sensor, e.g., a charge-coupled device (CCD) camera. Imagery generated by the optical sensor can be processed to determine detachment of analyte 270 from probe 260. For example, detachment of analyte 270 from probe 260 can result in a detectable change in electrical charge or pH. Sensor 400 can be configured to detect this change. In some embodiments, the method comprises using at least one sensor 400 configured to: (a) view the at least one event by an optical viewer; and (b) detect the at least one event by an optical sensor.

[0066] In some cases,, sensor 400 can detect disruption or detachment of bonds directly, indirectly, or both. In some cases, sensor 400 can be configured as an optical sensor, electromagnetic sensor, mechanical sensor, chemical sensor, or any combination thereof. For example, using methods herein to disrupt or detach bonds via force source 300 acting on force transmitter 280, sensor 400 can be configured to detect when some of force transmitter 280 having analyte 270 coupled thereto detach from probe 260. Such disruption or detachment can be used to determine molecular affinity between analyte 270 and probe 260. Methods herein can determine molecular affinity bycorrelating the applied force, e.g., strength and duration of the applied force, with the disruption or detachment event.Bond survival probability

[0067] System 100 can be configured to implement methods herein for determining or generating bond survival probability. In some embodiments, the method comprises aggregating the plurality of events to determine a statistical quality of data for each of the events. In some embodiments, each data for each of the events comprises a bond survival probability. In some embodiments, the bond survival probability comprises a dissociation chance of one or more molecular interactions between the capture moiety 260 and the analyte 270. In some embodiments, the identifying comprises determining a bond survival probability between the capture moiety 260 and the analyte 270 based on the at least one event. In some embodiments, each data for each of the events has one or more types of statistical quality determined at least from a cardinality of each data.

[0068] For example, as illustrated in FIGs. 4A-4C, systems and methods herein can be used to generate an energy landscape of molecular interaction for determining molecular affinity and an associated bond survival probability. In some cases, the energy landscape can be considered a fingerprint of molecular interaction and used to uniquely identify analyte 270. In some embodiments, the bond survival probability comprises a dissociation chance of one or more molecular interactions between the capture moiety 260 and the analyte 270. Bond survival probability can be useful for identifying or characterizing analyte 270, e.g., a biologies candidate such as an antibody. In some embodiments, the bond survival probability is used to identify the analyte 270 as a lead candidate drug for a disease.

[0069] Compared to other methods such as surface plasmon resonance (SPR) techniques, bond survival probability herein can be used to (i) obtain better (e.g., better accuracy or resolution) association and dissociation constants (konand koff) in less time and with fewer experiments and (ii) provide better and more useful measures of molecular interaction characteristics. In some cases, systems and methods herein can perform adaptive sampling of molecular complexes 800, which can be useful for understanding molecular affinity even with scarce data. For example, generating more data from measurements for a given applied force and duration force can improve the quality of the data; however, improving beyond a predetermined level of data quality may not be useful. Accordingly, compared to other methods, methods herein can use adaptive sampling to generate data sufficient for determining bond survival probability in a more efficient and less timeconsuming manner without sacrificing accuracy.

[0070] In some embodiments, the method comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of ranges of the time-varying force and (iii) theplurality of periods of time. For example, FIG. 4A depicts a surface plot of bond survival probability 701 versus applied force 702 and time 703 using systems and methods herein. Bond survival probability can range from 0 to 1 where 0 is a low probability of a bond survival (e.g., analyte 270 detaching from probe 260) and 1 is a high probability of bond survival (e.g., analyte 270 remaining attached to probe 260). Compared to other methods, methods herein can generate and use FIG. 4A to uniquely identify analyte 270 under a range of applied forces, duration of the applied forces, or loading rate of the applied forces. For example, other methods may only determine an “on or off’ state (e.g., detachment or no detachment between analyte 270 and probe 260), may determine the “on or off’ state using exhaustive studies of analyte concentrations such as SPR, or may determine the “on or off’ state using a single period of time under an applied force.

[0071] In some embodiments, the method comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of ranges of the time-varying force at a predetermined time of the plurality of periods of time. In some embodiments, the method comprises generating and displaying a plot of (i) each bond survival probability against (ii) the plurality of periods of time at a predetermined range or magnitude of the plurality of ranges of the time-varying force. For example, FIG. 4B depicts a line plot of bond survival probability 701 versus time 703 for different strengths of applied force 702, e.g., 56 piconewtons (pN, 702.1), 67 pN (702.2), 78 pN (702.3), and 89 pN (702.4) over a range of 0 to 10 seconds.

[0072] In some embodiments, the method comprises determining a bond survival probability between the capture moiety 260 and the analyte 270 based on the at least one event. In some embodiments, the method comprises generating a surface plot of (i) the bond survival probability against (ii) the magnetic field condition. In some embodiments, the method comprises extracting a characteristic time from the surface plot to determine (i) a diffusion relaxation time and (ii) dissociation constants. In some embodiments, the method comprises using the dissociation constants to identify the analyte as a lead candidate drug for a disease.

[0073] For example, FIG. 4C depicts characteristic times 720, t(f), versus applied force 702. In some cases, characteristic times 720, t(f), can range from about 10'3to 101s. As shown in Equations 1-3, from characteristic time, t(f), diffusion relaxation time, fo, can be determined to subsequently determine an association constant (kon) and a dissociation constant (koff) as functions of the energy landscape and fo.

[0074] In some embodiments, the method comprises dynamically updating the magnetic field condition in real time based at least on determining a statistical quality of data for each of the events. For example, with a moving and time varying force source 300, statistical events, such as molecular dissociation, can produce data that is of unknown quality a priori. In some cases, each dissociation event may not be deterministic so various features of a graph, such as FIG. 4A, can have differing underlying data contributing to the features. Thus, more data contributing to a feature in the graph, which can represent probabilities by gathering individual dissociation events of molecular bonds, may result in a statistically higher quality and vice versa. A natural variation of statistical quality in features of a graph, such as FIG. 4A, can arise depending on the amount of data that contributed to the feature. In some embodiments, updating the magnetic field condition comprises changing at least one parameter of the time-varying force. In some embodiments, updating the magnetic field condition comprises changing at least one parameter of the period of time. In some embodiments, updating the magnetic field condition comprises changing at least one parameter of the motion path. By such updates, methods herein can dynamically improve statistical quality of data to meet predetermined criteria.EXAMPLES

[0075] While various examples of the present disclosure have been shown and described herein, such examples are provided by way of example only. Numerous variations, changes, or substitutions may occur without departing from the present disclosure. It should be understood that various alternatives to the examples described herein may be employed.Example 1Computing systems

[0076] In an aspect, disclosed herein is a system comprising at least one processor and instructions executable by the at least one processor to cause the at least one processor to perform operations comprising: (a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; (b) using the capture moiety to capture the analyte; (c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and (d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii)-

[0077] In an aspect, disclosed herein is a computer-implemented method for identifying an analyte, the method comprising: (a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle; (b) using the capture moiety to capture the analyte; (c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and (d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

[0078] Referring to FIG. 7, a block diagram is shown depicting an exemplary machine that includes a computer system 700 (e.g., a processing or computing system) within which a set of instructions can execute for causing a device to perform or execute any one or more of the aspects and / or methodologies herein. The components in FIG. 7 are examples only and do not limit the scope of use or functionality of any hardware, software, embedded logic component, or a combination of two or more such components implementing particular embodiments.

[0079] Computer system 700 may include one or more processors 701, a memory 703, and a storage 708 that communicate with each other, and with other components, via a bus 740. The bus 740 may also link a display 732, one or more input devices 733 (which may, for example, include a keypad, a keyboard, a mouse, a stylus, etc.), one or more output devices 734, one or more storage devices 735, and various tangible storage media 736. All of these elements may interface directly or via one or more interfaces or adaptors to the bus 740. For instance, the various tangible storage media 736 can interface with the bus 740 via storage medium interface 726. Computer system 700 may have any suitable physical form, including but not limited to one or more integrated circuits (ICs), printed circuit boards (PCBs), mobile handheld devices (such as mobile telephones or PDAs), laptop or notebook computers, distributed computer systems, computing grids, or servers.

[0080] Computer system 700 includes one or more processor(s) 701 (e.g., central processing units (CPUs) or general purpose graphics processing units (GPGPUs)) that carry out functions. Processor(s) 701 optionally contains a cache memory unit 702 for temporary local storage of instructions, data, or computer addresses. Processor(s) 701 are configured to assist in execution of computer readable instructions. Computer system 700 may provide functionality for the components depicted in FIG. 7 as a result of the processor(s) 701 executing non-transitory,processor-executable instructions embodied in one or more tangible computer-readable storage media, such as memory 703, storage 708, storage devices 735, and / or storage medium 736. The computer-readable media may store software that implements particular embodiments, and processor(s) 701 may execute the software. Memory 703 may read the software from one or more other computer-readable media (such as mass storage device(s) 735, 736) or from one or more other sources through a suitable interface, such as network interface 720. The software may cause processor(s) 701 to carry out one or more processes or one or more steps of one or more processes described or illustrated herein. Carrying out such processes or steps may include defining data structures stored in memory 703 and modifying the data structures as directed by the software.

[0081] The memory 703 may include various components (e.g., machine readable media) including, but not limited to, a random access memory component (e.g., RAM 704) (e.g., static RAM (SRAM), dynamic RAM (DRAM), ferroelectric random access memory (FRAM), phasechange random access memory (PRAM), etc.), a read-only memory component (e.g., ROM 705), and any combinations thereof. ROM 705 may act to communicate data and instructions unidirectionally to processor(s) 701, and RAM 704 may act to communicate data and instructions bidirectionally with processor(s) 701. ROM 705 and RAM 704 may include any suitable tangible computer-readable media described below. In one example, a basic input / output system 706 (BIOS), including basic routines that help to transfer information between elements within computer system 700, such as during start-up, may be stored in the memory 703.

[0082] Fixed storage 708 is connected bidirectionally to processor(s) 701, optionally through storage control unit 707. Fixed storage 708 provides additional data storage capacity and may also include any suitable tangible computer-readable media described herein. Storage 708 may be used to store operating system 709, executable(s) 710, data 711, applications 712 (application programs), and the like. Storage 708 can also include an optical disk drive, a solid-state memory device (e.g., flash-based systems), or a combination of any of the above. Information in storage 708 may, in appropriate cases, be incorporated as virtual memory in memory 703.

[0083] In one example, storage device(s) 735 may be removably interfaced with computer system 700 (e.g., via an external port connector (not shown)) via a storage device interface 725. Particularly, storage device(s) 735 and an associated machine-readable medium may provide nonvolatile and / or volatile storage of machine-readable instructions, data structures, program modules, and / or other data for the computer system 700. In one example, software may reside, completely or partially, within a machine-readable medium on storage device(s) 735. In another example, software may reside, completely or partially, within processor(s) 701.

[0084] Bus 740 connects a wide variety of subsystems. Herein, reference to a bus may encompass one or more digital signal lines serving a common function, where appropriate. Bus 740 may be any of several types of bus structures including, but not limited to, a memory bus, a memory controller, a peripheral bus, a local bus, and any combinations thereof, using any of a variety of bus architectures. As an example and not by way of limitation, such architectures include an Industry Standard Architecture (ISA) bus, an Enhanced ISA (EISA) bus, a Micro Channel Architecture (MCA) bus, a Video Electronics Standards Association local bus (VLB), a Peripheral Component Interconnect (PCI) bus, a PCI-Express (PCI-X) bus, an Accelerated Graphics Port (AGP) bus, HyperTransport (HTX) bus, serial advanced technology attachment (SATA) bus, and any combinations thereof.

[0085] Computer system 700 may also include an input device 733. In one example, a user of computer system 700 may enter commands and / or other information into computer system 700 via input device(s) 733. Examples of an input device(s) 733 include, but are not limited to, an alphanumeric input device (e.g., a keyboard), a pointing device (e.g., a mouse or touchpad), a touchpad, a touch screen, a multi-touch screen, a joystick, a stylus, a gamepad, an audio input device (e.g., a microphone, a voice response system, etc.), an optical scanner, a video or still image capture device (e.g., a camera), and any combinations thereof. In some embodiments, the input device is a Kinect®, Leap Motion®, or the like. Input device(s) 733 may be interfaced to bus 740 via any of a variety of input interfaces 723 (e.g., input interface 723) including, but not limited to, serial, parallel, game port, USB, FIREWIRE, THUNDERBOLT, or any combination of the above.

[0086] In particular embodiments, when computer system 700 is connected to network 730, computer system 700 may communicate with other devices, specifically mobile devices and enterprise systems, distributed computing systems, cloud storage systems, cloud computing systems, and the like, connected to network 730. Communications to and from computer system 700 may be sent through network interface 720. For example, network interface 720 may receive incoming communications (such as requests or responses from other devices) in the form of one or more packets (such as Internet Protocol (IP) packets) from network 730, and computer system 700 may store the incoming communications in memory 703 for processing. Computer system 700 may similarly store outgoing communications (such as requests or responses to other devices) in the form of one or more packets in memory 703 and communicated to network 730 from network interface 720. Processor(s) 701 may access these communication packets stored in memory 703 for processing.

[0087] Examples of the network interface 720 include, but are not limited to, a network interface card, a modem, and any combination thereof. Examples of a network 730 or network segment 730include, but are not limited to, a distributed computing system, a cloud computing system, a wide area network (WAN) (e.g., the Internet, an enterprise network), a local area network (LAN) (e.g., a network associated with an office, a building, a campus or other relatively small geographic space), a telephone network, a direct connection between two computing devices, a peer-to-peer network, and any combinations thereof. A network, such as network 730, may employ a wired and / or a wireless mode of communication. In general, any network topology may be used.

[0088] Information and data can be displayed through a display 732. Examples of a display 732 include, but are not limited to, a cathode ray tube (CRT), a liquid crystal display (LCD), a thin film transistor liquid crystal display (TFT-LCD), an organic liquid crystal display (OLED) such as a passive-matrix OLED (PMOLED) or active-matrix OLED (AMOLED) display, a plasma display, and any combinations thereof. The display 732 can interface to the processor(s) 701, memory 703, and fixed storage 708, as well as other devices, such as input device(s) 733, via the bus 740. The display 732 is linked to the bus 740 via a video interface 722, and transport of data between the display 732 and the bus 740 can be controlled via the graphics control 721. In some embodiments, the display is a video projector. In some embodiments, the display is a head-mounted display (HMD) such as a VR headset. In further embodiments, suitable VR headsets include, by way of non-limiting examples, HTC Vive®, Oculus Rift®, Samsung Gear VR®, Microsoft HoloLens®, Razer OSVR®, FOVE VR®, Zeiss VR One®, Avegant Glyph®, Freefly VR® headset, and the like. In still further embodiments, the display is a combination of devices such as those disclosed herein.

[0089] In addition to a display 732, computer system 700 may include one or more other peripheral output devices 734 including, but not limited to, an audio speaker, a printer, a storage device, and any combinations thereof. Such peripheral output devices may be connected to the bus 740 via an output interface 724. Examples of an output interface 724 include, but are not limited to, a serial port, a parallel connection, a USB port, a FIREWIRE port, a THUNDERBOLT port, and any combinations thereof.

[0090] In addition or as an alternative, computer system 700 may provide functionality as a result of logic hardwired or otherwise embodied in a circuit, which may operate in place of or together with software to execute one or more processes or one or more steps of one or more processes described or illustrated herein. Reference to software in this disclosure may encompass logic, and reference to logic may encompass software. Moreover, reference to a computer-readable medium may encompass a circuit (such as an IC) storing software for execution, a circuit embodying logic for execution, or both, where appropriate. The present disclosure encompasses any suitable combination of hardware, software, or both.

[0091] Various illustrative logical blocks, modules, circuits, and algorithm steps described in connection with the embodiments disclosed herein may be implemented as electronic hardware, computer software, or combinations of both. To clearly illustrate this interchangeability of hardware and software, various illustrative components, blocks, modules, circuits, and steps have been described above generally in terms of their functionality.

[0092] The various illustrative logical blocks, modules, and circuits described in connection with the embodiments disclosed herein may be implemented or performed with a general purpose processor, a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic device, discrete gate or transistor logic, discrete hardware components, or any combination thereof designed to perform the functions described herein. A general purpose processor may be a microprocessor, but in the alternative, the processor may be any conventional processor, controller, microcontroller, or state machine. A processor may also be implemented as a combination of computing devices, e.g., a combination of a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors in conjunction with a DSP core, or any other such configuration.

[0093] The steps of a method or algorithm described in connection with the embodiments disclosed herein may be embodied directly in hardware, in a software module executed by one or more processor(s), or in a combination of the two. A software module may reside in RAM memory, flash memory, ROM memory, EPROM memory, EEPROM memory, registers, hard disk, a removable disk, a CD-ROM, or any other form of storage medium. An exemplary storage medium is coupled to the processor such the processor can read information from, and write information to, the storage medium. In the alternative, the storage medium may be integral to the processor. The processor and the storage medium may reside in an ASIC. The ASIC may reside in a user terminal. In the alternative, the processor and the storage medium may reside as discrete components in a user terminal.

[0094] In accordance with the description herein, suitable computing devices include, by way of non-limiting examples, server computers, desktop computers, laptop computers, notebook computers, sub-notebook computers, netbook computers, netpad computers, set-top computers, media streaming devices, handheld computers, Internet appliances, mobile smartphones, tablet computers, personal digital assistants, video game consoles, and vehicles. Select televisions, video players, and digital music players with optional computer network connectivity are suitable for use in the system described herein. Suitable tablet computers, in various embodiments, include those with booklet, slate, and convertible configurations.

[0095] In some embodiments, the computing device includes an operating system configured to perform executable instructions. The operating system is, for example, software, including programs and data, which manages the device’s hardware and provides services for execution of applications. Suitable server operating systems include, by way of non-limiting examples, FreeBSD®, OpenBSD®, NetBSD®, Linux®, Apple® Mac OS X Server®, Oracle Solaris®, Windows Server®, and Novell NetWare®. Suitable personal computer operating systems include, by way of non-limiting examples, Microsoft Windows®, Apple Mac® OS X, UNIX®, and UNIX-like operating systems such as GNU / Linux®. In some embodiments, the operating system is provided by cloud computing. Suitable mobile smartphone operating systems include, by way of nonlimiting examples, Nokia Symbian® OS, Apple® iOS, Research In Motion BlackBerry® OS, Google® Android®, Microsoft® Windows Phone® OS, Microsoft® Windows Mobile OS, Linux®, and Palm® WebOS. Suitable media streaming device operating systems include, by way of nonlimiting examples, Apple TV®, Roku®, Boxee®, Google TV®, Google Chromecast®, Amazon Fire®, and Samsung® HomeSync®. Suitable video game console operating systems include, by way of non-limiting examples, Sony® PS3®, Sony® PS4®, Microsoft® Xbox 360®, Microsoft Xbox One®, Nintendo Wii®, Nintendo Wii U®, and Ouya®. Suitable virtual reality headset systems include, by way of non-limiting example, Meta Oculus®.Non-transitory computer readable storage mediums

[0096] In some embodiments, the platforms, systems, media, and methods disclosed herein include one or more non-transitory computer readable storage media encoded with a program including instructions executable by the operating system of an optionally networked computing device. In further embodiments, a computer readable storage medium is a tangible component of a computing device. In still further embodiments, a computer readable storage medium is optionally removable from a computing device. In some embodiments, a computer readable storage medium includes, by way of non-limiting examples, CD-ROMs, DVDs, flash memory devices, solid state memory, magnetic disk drives, magnetic tape drives, optical disk drives, distributed computing systems including cloud computing systems and services, and the like. In some cases, the program and instructions are permanently, substantially permanently, semi -permanently, or non-transitorily encoded on the media.Computer programs

[0097] In some embodiments, the platforms, systems, media, and methods disclosed herein include at least one computer program, or use of the same. A computer program includes a sequence of instructions, executable by one or more processor(s) of the computing device’s CPU, written to perform a specified task. Computer readable instructions may be implemented as programmodules, such as functions, objects, Application Programming Interfaces (APIs), computing data structures, and the like, that perform particular tasks or implement particular abstract data types. In light of the disclosure provided herein, a computer program may be written in various versions of various languages.

[0098] The functionality of the computer readable instructions may be combined or distributed as desired in various environments. In some embodiments, a computer program comprises one sequence of instructions. In some embodiments, a computer program comprises a plurality of sequences of instructions. In some embodiments, a computer program is provided from one location. In other embodiments, a computer program is provided from a plurality of locations. In various embodiments, a computer program includes one or more software modules. In various embodiments, a computer program includes, in part or in whole, one or more web applications, one or more mobile applications, one or more standalone applications, one or more web browser plug-ins, extensions, add-ins, or add-ons, or combinations thereof.Web applications

[0099] In some embodiments, a computer program includes a web application. In light of the disclosure provided herein, a web application, in various embodiments, utilizes one or more software frameworks and one or more database systems. In some embodiments, a web application is created upon a software framework such as Microsoft® .NET or Ruby on Rails® (RoR). In some embodiments, a web application utilizes one or more database systems including, by way of nonlimiting examples, relational, non-relational, object oriented, associative, and XML database systems. In further embodiments, suitable relational database systems include, by way of nonlimiting examples, Microsoft® structured query language (SQL) Server, mySQL™, and Oracle®. A web application, in various embodiments, is written in one or more versions of one or more languages. A web application may be written in one or more markup languages, presentation definition languages, client-side scripting languages, server-side coding languages, database query languages, or combinations thereof. In some embodiments, a web application is written to some extent in a markup language such as Hypertext Markup Language (HTML), Extensible Hypertext Markup Language (XHTML), or extensible Markup Language (XML). In some embodiments, a web application is written to some extent in a presentation definition language such as Cascading Style Sheets (CSS). In some embodiments, a web application is written to some extent in a clientside scripting language such as Asynchronous Javascript and XML® (AJAX), Flash Actionscript, Javascript®, or Silverlight®. In some embodiments, a web application is written to some extent in a server-side coding language such as Active Server Pages® (ASP), ColdFusion®, Perl®, Java®, JavaServer Pages® (JSP), Hypertext Preprocessor® (PHP), Python®, Ruby®, Tel®, Smalltalk®,WebDNA®, or Groovy®. In some embodiments, a web application is written to some extent in a database query language such as Structured Query Language (SQL). In some embodiments, a web application integrates enterprise server products such as IBM Lotus Domino®. In some embodiments, a web application includes a media player element. In various further embodiments, a media player element utilizes one or more of many suitable multimedia technologies including, by way of non-limiting examples, Adobe® Flash®, HTML 5, Apple® QuickTime®, Microsoft Silverlight®, Java®, and Unity®.

[0100] Referring to FIG. 8, in a particular embodiment, an application provision system comprises one or more databases 800 accessed by a relational database management system (RDBMS) 810. Suitable RDBMSs include Firebird®, MySQL®, PostgreSQL®, SQLite®, Oracle Database®, Microsoft SQL Server®, IBM DB2®, IBM Informix®, SAP Sybase®, SAP Sybase®, Teradata®, PostGIS®, time-series databases, graph databases, and the like. In this embodiment, the application provision system further comprises one or more application severs 820 (such as Java® servers, .NET® servers, PHP® servers, and the like) and one or more web servers 830 (such as Apache®, IIS®, GWS® and the like). The web server(s) optionally expose one or more web services via app application programming interfaces (APIs) 840. Via a network, such as the Internet, the system provides browser-based and / or mobile native user interfaces.

[0101] Referring to FIG. 9, in a particular embodiment, an application provision system alternatively has a distributed, cloud-based architecture 900 and comprises elastically load balanced, auto-scaling web server resources 910 and application server resources 920 as well synchronously replicated databases 930.Mobile applications

[0102] In some embodiments, a computer program includes a mobile application provided to a mobile computing device. In some embodiments, the mobile application is provided to a mobile computing device at the time it is manufactured. In other embodiments, the mobile application is provided to a mobile computing device via the computer network described herein.

[0103] In view of the disclosure provided herein, a mobile application is created by techniques using hardware, languages, and development environments. Mobile applications are written in several languages. Suitable programming languages include, by way of non-limiting examples, C, C++, C#, Objective-C, Java®, Javascript®, Pascal®, Object Pascal®, Python™, Ruby®, VB.NET®, WML®, and XHTML / HTML with or without CSS, or combinations thereof.

[0104] Suitable mobile application development environments are available from several sources. Commercially available development environments include, by way of non-limiting examples, AirplaySDK®, alcheMo®, Appcelerator®, Celsius®, Bedrock®, Flash Lite®, .NET CompactFramework®, Rhomobile®, and WorkLight Mobile Platform®. Other development environments are available without cost including, by way of non-limiting examples, Lazarus®, MobiFlex®, MoSync®, and Phonegap®. Also, mobile device manufacturers distribute software developer kits including, by way of non-limiting examples, iPhone® and iPad® (iOS) SDK, Android® SDK, BlackBerry® SDK, BREW SDK, Palm® OS SDK, Symbian® SDK, webOS® SDK, and Windows® Mobile SDK.

[0105] Several commercial forums are available for distribution of mobile applications including, by way of non-limiting examples, Apple® App Store, Google® Play, Chrome® WebStore, BlackBerry® App World, App Store® for Palm devices, App Catalog® for webOS, Windows® Marketplace for Mobile, Ovi Store for Nokia® devices, Samsung® Apps, and Nintendo® DSi Shop. Standalone applications

[0106] In some embodiments, a computer program includes a standalone application, which is a program that is run as an independent computer process, not an add-on to an existing process, e.g., not a plug-in. Standalone applications are often compiled. A compiler is a computer program(s) that transforms source code written in a programming language into binary object code such as assembly language or machine code. Suitable compiled programming languages include, by way of non-limiting examples, C, C++, Objective-C®, COBOL®, Delphi®, Eiffel®, Java®, Lisp®, Python®, Visual Basic®, and VB .NET®, or combinations thereof. Compilation is often performed, at least in part, to create an executable program. In some embodiments, a computer program includes one or more executable compiled applications. Additionally, microservices related to Python® and JavaScript® may be used.Web browser plug-ins

[0107] In some embodiments, the computer program includes a web browser plug-in (e.g., web extension, etc.). In computing, a plug-in is one or more software components that add specific functionality to a larger software application. Makers of software applications support plug-ins to enable third-party developers to create abilities which extend an application, to support easily adding new features, and to reduce the size of an application. When supported, plug-ins enable customizing the functionality of a software application. For example, plug-ins are commonly used in web browsers to play video, generate interactivity, scan for viruses, and display particular file types. Several web browser plug-ins may include Adobe Flash Player®, Microsoft Silverlight®, and Apple QuickTime®. In some embodiments, the toolbar comprises one or more web browser extensions, add-ins, or add-ons. In some embodiments, the toolbar comprises one or more explorer bars, tool bands, or desk bands.

[0108] In view of the disclosure provided herein, several plug-in frameworks are available that enable development of plug-ins in various programming languages, including, by way of nonlimiting examples, C++, Delphi®, Java®, PHP®, Python®, and VB .NET®, or combinations thereof.

[0109] Web browsers (also called Internet browsers) are software applications, designed for use with network-connected computing devices, for retrieving, presenting, and traversing information resources on the World Wide Web. Suitable web browsers include, by way of non-limiting examples, Microsoft Internet Explorer®, Mozilla Firefox®, Google Chrome®, Apple Safari®, Opera Software Opera®, and KDE Konqueror®. In some embodiments, the web browser is a mobile web browser. Mobile web browsers (also called microbrowsers, mini-browsers, and wireless browsers) are designed for use on mobile computing devices including, by way of non-limiting examples, handheld computers, tablet computers, netbook computers, subnotebook computers, smartphones, music players, personal digital assistants (PDAs), and handheld video game systems. Suitable mobile web browsers include, by way of non-limiting examples, Google Android® browser, RIM BlackBerry® Browser, Apple Safari®, Palm Blazer®, Palm WebOS® Browser, Mozilla Firefox® for mobile, Microsoft Internet Explorer Mobile®, Amazon Kindle Basic Web®, Nokia Browser®, Opera Software Opera Mobile®, and Sony PSP® browser.Software modules

[0110] In some embodiments, the platforms, systems, media, and methods disclosed herein include software, server, and / or database modules, or use of the same. In view of the disclosure provided herein, software modules are created by techniques using machines, software, and languages. The software modules disclosed herein are implemented in a multitude of ways. In various embodiments, a software module comprises a file, a section of code, a programming object, a programming structure, or combinations thereof. In further various embodiments, a software module comprises a plurality of files, a plurality of sections of code, a plurality of programming objects, a plurality of programming structures, or combinations thereof. In various embodiments, the one or more software modules comprise, by way of non-limiting examples, a web application, a mobile application, and a standalone application. In some embodiments, software modules are in one computer program or application. In other embodiments, software modules are in more than one computer program or application. In some embodiments, software modules are hosted on one machine. In other embodiments, software modules are hosted on more than one machine. In further embodiments, software modules are hosted on a distributed computing platform such as a cloud computing platform. In some embodiments, software modules are hosted on one or more machines in one location. In other embodiments, software modules are hosted on one or more machines in more than one location.Databases[OHl] In some embodiments, the platforms, systems, media, and methods disclosed herein include one or more databases (DB), or use of the same. In view of the disclosure provided herein, many databases are suitable for storage and retrieval data. In various embodiments, suitable databases include, by way of non -limiting examples, relational databases, non-relational databases, object oriented databases, object databases, entity-relationship model databases, associative databases, XML databases, time-series databases, graph databases, and the like. Further non-limiting examples include SQL, PostgreSQL®, MySQL®, Oracle®, DB2®, and Sybase. In some embodiments, a database is internet-based. In further embodiments, a database is web-based. In still further embodiments, a database is cloud computing-based. In a particular embodiment, a database is a distributed database. In other embodiments, a database is based on one or more local computer storage devices.Terms and Definitions

[0112] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this present disclosure belongs.

[0113] As used herein, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Any reference to “or” herein is intended to encompass “and / or” unless otherwise stated.

[0114] As used herein, the term “about” in some cases refers to an amount that is approximately the stated amount.

[0115] As used herein, the term “about” refers to an amount that is near the stated amount by 10%, 5%, or 1%, including increments therein.

[0116] As used herein, the term “about” in reference to a percentage refers to an amount that is greater or less the stated percentage by 10%, 5%, or 1%, including increments therein.

[0117] As used herein, the phrases “at least one,” “one or more,” and “and / or” are open-ended expressions that are both conjunctive and disjunctive in operation. For example, each of the expressions “at least one of A, B and C,” “at least one of A, B, or C,” “one or more of A, B, and C”, “one or more of A, B, or C” and “A, B, and / or C” means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B and C together.

[0118] While preferred embodiments of the present disclosure have been shown and described herein, such embodiments are provided by way of example only. It is not intended that the present disclosure be limited by the specific examples provided within the specification. While the present disclosure has been described with reference to the aforementioned specification, the descriptions and illustrations of the embodiments herein are not meant to be construed in a limiting sense.Numerous variations, changes, and substitutions may occur without departing from the present disclosure. Furthermore, it shall be understood that all aspects of the present disclosure are not limited to the specific depictions, configurations, or relative proportions set forth herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the present disclosure described herein may be employed in practicing the present disclosure. It is therefore contemplated that the present disclosure shall also cover any such alternatives, modifications, variations, or equivalents. It is intended that the following claims define the scope of the present disclosure and that systems, methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method for identifying an analyte, the method comprising:(a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically- attractable particle;(b) using the capture moiety to capture the analyte;(c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and(d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

2. The method of claim 1, wherein the at least one event comprises a dissociation event between the capture moiety and the analyte.

3. The method of claim 1, further comprising dynamically updating the magnetic field condition in real time based at least on determining a statistical quality of data for each of the events.

4. The method of claim 3, wherein updating the magnetic field condition comprises changing at least one parameter of the time-varying force.

5. The method of claim 3, wherein updating the magnetic field condition comprises changing at least one parameter of the at least one period of time.

6. The method of claim 3, wherein updating the magnetic field condition comprises changing at least one parameter of the at least one motion path.

7. The method of claim 1, further comprising correlating the at least one event to at least one of (i) the time-varying force, (ii) the at least one period of time, or (iii) the at least one motion path.

8. The method of claim 1, further comprising selecting the magnetic field condition to have (i) a plurality of ranges of the time-varying force applied over (ii) a plurality of periods of time, (iii) a plurality of motion paths, or (iv) any combination of (i) - (iii).

9. The method of claim 8, further comprising:(a) applying the magnetic field having the magnetic field condition;(b) detecting a plurality of events caused by applying the magnetic field; and(c) aggregating the plurality of events to determine a statistical quality of data for each of the events, wherein each data for each of the events comprises a bond survival probability.

10. The method of claim 9, wherein each data for each of the events has one or more types of statistical quality determined at least from a cardinality of each data.

11. The method of claim 9, further comprising generating and displaying a plot of (i) each bond survival probability from each data against (ii) the plurality of ranges of the time-varying force at a predetermined time of the plurality of periods of time.

12. The method of claim 9, further comprising generating and displaying a plot of (i) each bond survival probability from each data against (ii) the plurality of periods of time at a predetermined range or magnitude of the plurality of ranges of the time-varying force.

13. The method of claim 9, further comprising generating and displaying a plot of (i) each bond survival probability from each data against (ii) the plurality of ranges of the time-varying force and (iii) the plurality of periods of time.

14. The method of claim 8, further comprising using the at least one motion path to generate the plurality of ranges of the time-varying force.

15. The method of claim 8, further comprising generating the plurality of periods of time by:(a) using the at least one motion path, wherein each period of time comprises one or more velocity profiles or one or more acceleration profiles of a movement of a magnetic source used to generate the magnetic field; or(b) using one or more excitation profiles of the magnetic source, wherein each excitation profile comprises one or more magnetic strength profiles or one or more magnetic strength duration profiles.

16. The method of claim 1, further comprising:(a) determining a bond survival probability between the capture moiety and the analyte based on the at least one event;(b) generating a surface plot of (i) the bond survival probability against (ii) the magnetic field condition;(c) extracting a characteristic time from the surface plot to determine (i) a diffusion relaxation time and (ii) dissociation constants; and(d) using the dissociation constants to identify the analyte as a lead candidate drug for a disease.

17. The method of claim 16, wherein the bond survival probability comprises a dissociation chance of one or more molecular interactions between the capture moiety and the analyte.

18. The method of claim 1, wherein applying the magnetic field comprises acting on the magnetically-attractable particle using the time-varying force.

19. The method of claim 18, wherein the time varying force comprises a peak force or a root mean square (rms) force of at least about 1 piconewton (pN), 10 pN, 100 pN, 1000 pN, or greater.

20. The method of claim 1, wherein applying the magnetic field comprises acting on the magnetically-attractable particle using the at least one period of time.

21. The method of claim 20, wherein the at least one period of time comprises at most about 1000 microseconds (ps), 100 ps, 10 ps, or less.

22. The method of claim 1, wherein applying the magnetic field comprises acting on the magnetically-attractable particle using the at least one motion path relative to the magnetically- attractable particle.

23. The method of claim 22, wherein the at least one motion path is (i) along a vertical axis of the magnetically-attractable particle, (ii) along a vertical axis spatially separated from the vertical axis in (i), (iii) along a horizontal plane parallel to the surface, or (iv) any combination of (i) - (iii).

24. The method of claim 22, wherein the at least one motion path is a linear motion path or a nonlinear motion path comprising a path distance of about 0 mm to 100 mm.

25. The method of claim 1, further comprising applying the magnetic field to the magnetically- attractable particle by using an external or an integrated (i) magnetic source, (ii) electromagnetic source, or (iii) a combination of (i) and (ii).

26. The method of claim 1, further comprising applying a force to the force transmitter by using an external or an integrated (i) electrokinetic source, (ii) dielectrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v).

27. The method of claim 1, wherein the capture moiety comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii).

28. The method of claim 1, wherein the analyte comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii).

29. The method of claim 1, wherein the force transmitter comprises at least one particle configured to couple to at least one analyte, and wherein the at least one particle comprises (i) a magnetic bead, (ii) a dielectric bead, (iii) a polystyrene bead, (iv) a charged molecule, or (v) any combination (i) - (iv).

30. The method of claim 1, wherein the force transmitter comprises (i) a nucleic acid, (ii) a secondary antibody coupled to a nucleic acid, (iii) a secondary antibody coupled to a magnetic bead, (iv) a secondary antibody coupled to a dielectric bead, (v) a secondary antibody coupled to a polystyrene bead, (vi) a bead modified with at least one fluorescent molecule, or (vii) any combination (i) - (vi).

31. The method of claim 1, further comprising coupling the capture moiety to the surface using a covalent bond or a non-covalent bond.

32. The method of claim 1, further comprising directing the solution to at least one array portion of the surface, wherein each array portion comprises at least 1, 10, 100, or more individually programmable portions.

33. The method of claim 32, wherein each individually programmable portion comprises at least 1, 10, 100, 1000, or more individual programmable assay areas.

34. The method of claim 33, wherein each individually programmable assay area comprises a surface area of at least about 0.01 square millimeters (mm2), 0.1 mm2, 1.0 mm2, 10 mm2, 100 mm , or greater.

35. The method of claim 1, further comprising using at least one sensor configured to:(a) view the at least one event by an optical viewer; and(b) detect the at least one event by an optical sensor.

36. The method of claim 35, wherein the at least one sensor comprises (i) the optical sensor, (ii) an electromagnetic sensor, (iii) a mechanical sensor, (iv) a chemical sensor, or (v) any combination of (i) - (iv).

37. The method of claim 1, further comprising:(a) directing, by a flow cell, the solution having the analyte to a reservoir proximate to the surface; and(b) suspending, by a top surface operatively coupled to the reservoir, the solution in a volume of the reservoir.

38. The method of claim 1, further comprising coupling the capture moiety to the surface by flowing a gas phase or a liquid phase to deposit ((3 -Aminopropyl) trimethoxy silane) (ATPMS), ((3 -Aminopropyl)tri ethoxy silane) (ATPES), or ((3-Glycidyloxypropyl) trimethoxy silane) (GOPS).

39. A cartridge for identifying an analyte, the cartridge comprising: a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically-attractable particle;a reservoir configured to suspend a solution proximate to the surface, wherein the solution comprises the analyte coupled to the magnetically-attractable particle; array portions of the surface configured to assay the analyte under application of a magnetic field, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and at least one sensor configured to detect at least one event of the analyte caused by the application of the magnetic field having the magnetic field condition, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

40. The cartridge of claim 39, wherein at least one event comprises a dissociation event between the capture moiety and the analyte.

41. The cartridge of claim 39, wherein the identifying comprises determining a bond survival probability between the capture moiety and the analyte based on the at least one event.

42. The cartridge of claim 41, wherein the bond survival probability comprises a dissociation chance of one or more molecular interactions between the capture moiety and the analyte.

43. The cartridge of claim 41, wherein the bond survival probability is used to identify the analyte as a lead candidate drug for a disease.

44. The cartridge of claim 39, wherein the magnetic field is configured to act on the magnetically-attractable particle with the time-varying force.

45. The cartridge of claim 44, wherein the time varying force comprises a peak force or a root mean square (rms) force of at least about 1 piconewton (pN), 10 pN, 100 pN, 1000 pN, or greater.

46. The cartridge of claim 39, wherein the magnetic field is configured to act on the magnetically-attractable particle over the at least one period of time.

47. The cartridge of claim 46, wherein the at least one period of time comprises at most about 1000 microseconds (ps), 100 psec, 10 psec, 1 psec, or less.

48. The cartridge of claim 39, wherein the magnetic field is configured to act on the magnetically-attractable particle over the at one least motion path relative to the magnetically- attractable particle.

49. The cartridge of claim 48 wherein the at least one motion path is (i) along a vertical axis of the magnetically-attractable particle, (ii) along a vertical axis spatially separated from the vertical axis in (i), (iii) along a horizontal plane parallel to the surface, or (iv) any combination of (i) - (iii).

50. The cartridge of claim 48, wherein the at least one motion path is a linear motion path or a nonlinear motion path comprising a path distance of about 0 mm to 100 mm.

51. The cartridge of claim 39, wherein the cartridge is configured to apply the magnetic field to the magnetically-attractable particle by coupling to an external or an integrated (i) magnetic source, (ii) electromagnetic source, or (iii) a combination of (i) and (ii).

52. The cartridge of claim 39, wherein the cartridge is configured to apply a force to the force transmitter by coupling to an external or an integrated (i) electrokinetic source, (ii) di electrophoretic source, (iii) optical source, (iv) acoustic source, (v) mechanical source, or (vi) any combination of (i) - (v).

53. The cartridge of claim 39, wherein the capture moiety comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii).

54. The cartridge of claim 39, wherein the analyte comprises (i) nucleic acids, (ii) polypeptides, (iii) proteins, (iv) antibodies, (v) viruses, (vi) toxins, (vii) cells, (viii) bacteria, or (ix) any combination of (i) - (viii).

55. The cartridge of claim 39, wherein the force transmitter comprises at least one particle configured to couple to at least one analyte, and wherein the at least one particle comprises (i) a magnetic bead, (ii) a dielectric bead, (iii) a polystyrene bead, (iv) a charged molecule, or (v) any combination (i) - (iv).

56. The cartridge of claim 39, wherein the force transmitter comprises (i) a nucleic acid, (ii) a secondary antibody coupled to a nucleic acid, (iii) a secondary antibody coupled to a magnetic bead, (iv) a secondary antibody coupled to a dielectric bead, (v) a secondary antibody coupled to a polystyrene bead, (vi) a bead modified with at least one fluorescent molecule, or (vii) any combination (i) - (vi).

57. The cartridge of claim 39, wherein the capture moiety is coupled to the surface using a covalent bond or a non-covalent bond.

58. The cartridge of claim 39, wherein at least one array portion of the surface comprises at least 1, 10, 100, or more individually programmable portions.

59. The cartridge of claim 58, wherein each individually programmable portion comprises at least 1, 10, 100, 1000, or more individual programmable assay areas.

60. The cartridge of claim 59, wherein each individually programmable assay area comprises a surface area of at least 0.01 square millimeters (mm2), 0.1 mm2, 1.0 mm2, 10 mm2, 100 mm2, or greater.

61. The cartridge of claim 39, wherein the at least one sensor comprises:an optical viewer configured to optically view the at least one event; and an optical sensor configured to optically detect the at least one event.

62. The cartridge of claim 39, wherein the at least one sensor comprises (i) the optical sensor, (ii) an electromagnetic sensor, (iii) a mechanical sensor, (iv) a chemical sensor, or (v) any combination of (i) - (iv).

63. The cartridge of claim 39, further comprising: a flow cell configured to direct the solution having the analyte to the reservoir proximate to the surface; and a top surface operatively coupled to the reservoir configured to suspend the solution in a volume of the reservoir; and64. The cartridge of claim 39, wherein coupling the capture moiety to the surface comprises flowing a gas phase or a liquid phase to deposit ((3 -Aminopropyl) trimethoxy silane) (ATPMS), ((3 -Aminopropyl)tri ethoxy silane) (ATPES), or ((3-Glycidyloxypropyl) trimethoxy silane) (GOPS).

65. The cartridge as in any of claims 39 to 64, wherein the cartridge is configured to couple to an instrument for identifying the analyte.

66. A system comprising at least one processor and instructions executable by the at least one processor to cause the at least one processor to perform operations comprising:(a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically- attractable particle;(b) using the capture moiety to capture the analyte;(c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and(d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

67. A computer-implemented method for identifying an analyte, the method comprising:(a) directing a solution having the analyte to a surface having a capture moiety coupled thereto, wherein the analyte is coupled to a force transmitter comprising a magnetically- attractable particle;(b) using the capture moiety to capture the analyte;(c) subsequent to (b), applying a magnetic field to the magnetically-attractable particle, wherein the magnetic field has a magnetic field condition that is sufficient to direct the analyte and the magnetically-attractable particle coupled thereto away from the capture moiety; and(d) using the magnetic field condition in (c) to identify at least one event of the analyte, wherein the magnetic field condition comprises (i) a time-varying force, (ii) at least one period of time, (iii) at least one motion path, or (iv) any combination of (i) - (iii).

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