Methods and systems for separating and manipulating a sample comprising a plurality of components

Abstract

A system for separating and manipulating a sample includes an apparatus comprising a substrate defining a sample surface for receiving a medium The apparatus also includes a first manipulation assembly operable to apply a force to a sample to cause the sample to migrate along a first pathway, and a second manipulation assembly operable to apply a force to a sample to cause at least a part of the sample to migrate along a second pathway. The second pathway is adjacent to or intersects the first pathway. Migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components. At least one of the first and second manipulation assembly is configured so as to define an individually controllable region along a part of the respective first and / or second pathway.

Description

METHODS AND SYSTEMS FOR SEPARATING AND MANIPULATING A SAMPLE COMPRISING A PLURALITY OF COMPONENTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from U.S. Application No.63 / 548, 695, titled SYSTEMS, APPARATUS AND METHODS FOR MULTIDIMENSIONAL MOVEMENT OF AT LEAST ONE COMPONENT, filed 01 February 2024, which is hereby incorporated by reference in its entirety.FIELD OF THE DISCLOSURE

[0002] This disclosure relates to methods and systems for separating and manipulating a sample comprising a plurality of components.BACKGROUND

[0003] Separation techniques have been widely used across various scientific and industrial fields for analysing complex mixtures and isolating specific components based on their physical or chemical properties. Examples include techniques such as electrophoresis, chromatography, and membranebased filtration. While these methods have proven valuable, they often face limitations that can hinder broader adoption and effectiveness.SUMMARY

[0004] According to a first aspect of the present disclosure, a system for separating and manipulating a sample comprising a plurality of components is provided. The system comprises: a substrate defining a sample surface for receiving a medium in which the sample can be received; a first manipulation assembly operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway; and a second manipulation assembly operable to apply a force to a sample to cause the at least a part of the sample to migrate within medium provided at the sample surface along a second pathway, wherein the second manipulation assembly is arranged so that the second pathway is adjacent to or intersects the first pathway. At least one of the first manipulation assembly and the second manipulation assembly is configured so as to define at least one individually controllable region along a part of the respective first and / or second pathway in which a first local force can be selectively applied within the individually controllable region so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region. The system further comprises a control unit configured to operate: the first manipulation assembly so as to cause the sample to migrate within medium provided at the sample surface along the first pathway; and the second manipulation assembly so as to cause at least a part of the sample to migrate within medium provided at the sample surface along the second pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components. The control unit is further configured to operate at least one of the first manipulation assembly and the second manipulation assembly to generate the first local forcewithin the respective first and / or second pathway so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region.

[0005] According to a second aspect of the present disclosure, a method of separating and manipulating a sample comprising a plurality of components is provided. The method comprises providing a medium comprising a sample, the sample comprising a plurality of components; applying a force to the sample so as to cause the sample to migrate along a first pathway through the medium; and applying a force to at least a part of the sample so as to cause the part of the sample to migrate from the first pathway along a second pathway through the medium, wherein the second pathway is adjacent to or intersects the first pathway. Migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components. An individually controllable region is provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a first local force to the individually controllable region so as to influence or cause migration of at least a part of sample migrate along the respective first and / or second pathway within the individually controllable region.BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The present disclosure will now be described in more detail with reference to the accompanying drawings, which are not intended to be limiting:

[0007] FIG. 1 depicts a block diagram of a system for separating and manipulating samples, according to aspects of the present disclosure.

[0008] FIGS. 2A to 2C depict schematic top views of an apparatus for manipulating and separating samples for use in the system of FIG. 1 .

[0009] FIGS. 3 A and 3C provide graphs depicting different electric field profiles for sample manipulation, according to aspects of the present disclosure and FIG. 3B provides a graph depicting voltage applied across electrodes of a first manipulation assembly over time.

[0010] FIG. 4 depicts a block diagram of a system for separating and manipulating samples, according to aspects of the present disclosure.

[0011] FIG. 5 A depicts a schematic top view of an apparatus for manipulating and separating samples for use in the system of FIG. 4 and FIG. 5B depicts a schematic cross-sectional view of an apparatus for manipulating and separating samples for use in the system of FIG. 4.

[0012] FIG. 6 depicts a schematic top view of an apparatus for manipulating and separating samples.

[0013] FIG. 7 depicts a part of the apparatus of FIG.6 in more detail.

[0014] FIGS. 8A and 8B depict block diagrams of systems for separating and manipulating samples, according to aspects of the present disclosure.

[0015] FIG. 9 provides a schematic perspective view of system for separating and manipulating samples, according to aspects of the present disclosure.

[0016] FIG. 10A and 10B schematically depict parts of an apparatus which can be used in systems and methods according to the disclosure.

[0017] FIG. 11 illustrates a flowchart of a method according to the disclosure.DETAILED DESCRIPTION

[0018] According to a first aspect of the present disclosure, a system for separating and manipulating a sample comprising a plurality of components is provided. The system comprises: a substrate defining a sample surface for receiving a medium in which the sample can be received; a first manipulation assembly operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway; and a second manipulation assembly operable to apply a force to a sample to cause the at least a part of the sample to migrate within medium provided at the sample surface along a second pathway, wherein the second manipulation assembly is arranged so that the second pathway is adjacent to or intersects the first pathway. At least one of the first manipulation assembly and the second manipulation assembly is configured so as to define at least one individually controllable region along a part of the respective first and / or second pathway in which a first local force can be selectively applied within the individually controllable region so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region. The system further comprises a control unit configured to operate: the first manipulation assembly so as to cause the sample to migrate within medium provided at the sample surface along the first pathway; and the second manipulation assembly so as to cause at least a part of the sample to migrate within medium provided at the sample surface along the second pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components. The control unit is further configured to operate at least one of the first manipulation assembly and the second manipulation assembly to generate the first local force within the respective first and / or second pathway so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region.

[0019] According to a second aspect of the present disclosure, a method of separating and manipulating a sample comprising a plurality of components is provided. The method comprises providing a medium comprising a sample, the sample comprising a plurality of components; applying a force to the sample so as to cause the sample to migrate along a first pathway through the medium; and applying a force to at least a part of the sample so as to cause the part of the sample to migrate from the first pathway along a second pathway through the medium, wherein the second pathway is adjacent to or intersects the first pathway. Migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components. An individually controllable region is provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a first local force to the individuallycontrollable region so as to influence or cause migration of at least a part of sample migrate along the respective first and / or second pathway within the individually controllable region.

[0020] Such systems and methods provide a means for separating and manipulating complex mixtures in an improved manner. That is, the systems and method allow for multi-dimensional manipulation of samples comprising plural components and separation in at least one of these dimensions. Moreover, the systems and methods use local control within at least one individually controllable region provided along at least one pathway to provide improved resolution and control over the manipulation and separation processes compared to traditional single and multi-dimensional separation techniques.

[0021] The provision of at least one the individually controllable region in which a local force can be provided allows for the manipulation of samples or components thereof to a greater degree . This can be used to provide more complex separations and a more precise level of control.

[0022] Traditional separation systems where manipulation of the samples is limited to movement under a single force along a single pathway are limited in the resolution and separation they provide. Further, these processes can be susceptible to unexpected variations (e.g. caused by the sample, contaminants or the medium) and subsequent handling is difficult. Further, many conventional separation approaches rely heavily on manual operation (e.g. due to non-integrated steps and the need for manual interventions), provide limited control over the separation process and subsequent or prior manipulation and have limitations such as sample size, speed and cost.

[0023] This can lead to issues with reproducibility and precision. The disclosed systems and method incorporating an individually controllable region allow for individual manipulation of components of the sample before, during and / or after separation. For example, after or during separation of the sample into components, those components within the individually controllable region can be manipulated separately to the other components. This can be used to move these components to a particular region (e.g. a sensing element or for better alignment with a subsequent pathway), to collect the component at a particular point (for example, to concentrate the sample) or to create a greater degree of separation between components. One of the primary challenges associated with molecular separation techniques is achieving optimal resolution. Resolution is critical for the accurate identification and analysis of molecular components, but several factors can interfere with the separation process, leading to overlap or poor differentiation between similar molecules. For example, interactions between molecules, variations in buffer conditions, or inconsistencies in the separation medium can all contribute to suboptimal resolution. The additional control provided herein can reduce these issues.

[0024] Further, the individually controllable region allows for the generation of more complex (i.e. non-uniform) and sophisticated force profiles across the respective pathway to create a greater degree of separation between components. For example, with the individually controllable region capable of providing a different force to the remainder of the pathway. The fine control over the local force within this region may also allow for adaptive separation protocols, where force profiles are adjusted in realtime, for example based on detected sample properties or separation progress.

[0025] The individually controllable region as well as the presence of plural pathways (i.e. the presence of at least a first and second pathway) can be used for manipulation of the sample or the components thereof before and / or after separation. This can be used for e.g. alignment of the sample or component with a particular region or transfer of the separated components for subsequent analysis steps using the same system used for the separation. Molecular separation techniques typically involve upstream preparation steps and downstream analysis steps. The systems and methods disclosed herein can reduce or avoid manual intervention, reduce complexity and help to preserve sample integrity.

[0026] Accordingly, there is provided a system which comprises a device which defines or comprises a first channel or flow path (when medium is provided on or in the device) and a second channel or flow path. A sample can be caused by a first actuator of the device to migrate along the first channel under the action of a first force imparted on the sample by the first actuator. The sample can be subsequently caused to migrate along the second channel under the action of a second force imparted on the sample (or a part thereof) by the second actuator. By selection of the first and / or second actuator and the medium, the device can cause the various components of sample to be separated out along at least one of the first or second routes (e.g. channels) through the medium. Further, at least one of the first or second actuator has an independently operable local actuator configured to generate a separate localised force within a portion of one of the first or second channels, which local actuator can generate a localised force to influence sample movement in that respect portion. There may be a plurality of individually or separately independent regions along the respective channel. The individually controllable regions may be discrete regions, for example adjacent or abutting regions, and / or may be overlapping regions.

[0027] Each of the first manipulation assembly and / or the second manipulation assembly may be configured to provide an individually controllable region. Where there are additional pathways and manipulation assemblies, such as a third, these too may be configured to provide an individually controllable region or a plurality of individually controllable regions.

[0028] The control unit may be configured to operate at least one of the first manipulation assembly and / or the second manipulation assembly so as to provide different force profiles along the respective first pathway and / or second pathway using the local force. These may be applied simultaneously or at different times. Similarly, in the method, applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium may comprise generating different force profiles along the respective first pathway and / or second pathway using the plurality of individually controllable regions. Examples might include generation of forces across the length of the respective pathway so as to provide a non-linear overall force, such as stepped forces, a force gradient, or any other non-linear profile.

[0029] By “influence or cause migration” of at least a part of sample along the respective first and / or second pathway within the individually controllable region”, it is meant that the sample or a part / portion thereof can be caused to migrate under the local force, which is different to the force(s) which can beapplied along the remainder of the pathway. This may be that the first local force may be applied while no force is applied to at least a part of the remainder of the respective pathway. Or it may have a different polarity, strength or intensity to at least a part of the remainder of the respective pathway. For example, where the respective manipulation assembly generates a first force across the entire pathway to cause the migration (for example, there is a first electrode set for generating a force along the extent of the pathway), excluding the first local force, the local force may influence the migration, for example by providing an opposing force or complimentary force within the individually controllable region, or by retaining a part of the sample therein. The local force applied within the individually controllable region may influence migration in the direction along the respective pathway.

[0030] The pathway having the individually controllable region(s) may further be the pathway along which the sample is separated. Having additional control over a separation can lead to better separation profiles, as set out above.

[0031] At least one of the first manipulation assembly and / or the second manipulation assembly may be configured to provide a plurality of individually controllable regions along a part of the respective first and / or second pathway in which a respective local force can be selectively applied to at least a part of sample within the respective individually controllable regions. In other words, there may be a plurality of individually or separately actuatable regions along the respective pathway, for example defined by different combinations of electrodes configured to produce a force within the individually controllable regions. The individually controllable regions may be discrete regions, for example adjacent or abutting regions, and / or may be overlapping regions. The control unit may be configured to operate the first manipulation assembly and / or the second manipulation assembly so as to apply a local force to each of the plurality of individually controllable regions. This can provide further precision and control over the manipulation and separation process, allowing for more targeted manipulation of specific sample components. In some embodiments, each of the first manipulation assembly and / or the second manipulation assembly may be configured to provide a plurality of individually controllable regions. Where there are additional pathways and manipulation assemblies, such as a third, these too may be configured to provide a plurality of individually controllable regions.

[0032] The control unit may be configured to operate at least one of the first manipulation assembly and / or the second manipulation assembly so as to provide a different force profiles along the respective first pathway and / or second pathway using the respective local forces in the plurality of individually controllable regions. Accordingly, there may be at least a first profile across a first individually controllable region and at least a second profile across a second individually controllable region. Where these do not encompass the entire respective pathway, there may be additional force profiles across the remaining regions of the pathway. The force profiles may be a positive, negative or a zero force value. These may be applied simultaneously or at different times. This enables the creation of complex force gradients along the pathways, allowing for more sophisticated separation and manipulation of sample components.

[0033] Accordingly, the method disclosed herein may include providing a plurality of individually controllable regions along at least one of the first pathway and / or the second pathway; and wherein “applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium” comprises selectively applying a local force to each of the individually controllable regions so as to cause a part of the sample to migrate along the respective first and / or second pathway within each of the plurality of individually controllable region. Applying a force so as to cause the sample to migrate along a first pathway according to the methods disclosed herein may comprise applying a plurality of forces across the first pathway such that the force applied to the sample varies across the first pathway. The plurality of force profiles may comprise a first local force across the individually controllable region of the first pathway and a second local force across a second portion of the first pathway. The method may additionally or alternatively comprise applying a first force profile across a first portion of the second pathway and applying a second force profile across a second portion of the second pathway such that the force applied varies across the second pathway. In such a configuration, the second manipulation assembly may comprise a plurality of electrodes operable to provide the plurality of force profiles across the second pathway and wherein at least one electrode of the plurality of electrodes is located along the second pathway. For example, the second manipulation assembly may comprise a first electrode set comprising at least two electrodes of the plurality of electrodes arranged to provide a force at the first portion and a second electrode set comprising at least two electrodes of the plurality of electrodes operable to provide a force at the second portion.Separation and Manipulation assemblies

[0034] Migration along at least one of the first pathway and / or the second pathway is arranged to (i.e. is so as to) separate the sample into a plurality of components.

[0035] The first manipulation assembly and second manipulation assembly may each comprise a first electrode set arranged to define the respective first pathway and second pathway and to provide the respective force. Use of electrodes provides a precise control over the applied forces and allows for integration of the assemblies into a chip or integrated circuit for precise manufacture and miniaturisation. The methods disclosed herein may further comprise providing a first manipulation assembly operable to provide a force to the sample to cause the sample to migrate along the first pathway; and a second manipulation assembly operable to apply a force to the sample to cause at least a part of the sample to migrate along the second pathway.

[0036] For example, the systems and methods can use these electrodes to generate electric or magnetic fields that guide sample migration along predefined paths or affect a change in the environmental conditions which in turn create a force which causes movement of the along a pathway.

[0037] The first electrode set can comprise a pair of electrodes provided as an electrically connectable or connected pair of field-generating electrodes for generating a field for influencing the sample or a part thereof (e.g. an electric field and / or a magnetic field). Each of the pair of electrodes can be providedat an opposing end of the pathway and to define the entire length of the pathway. In other words, these are spaced apart and medium is provided therebetween so that the region defined between them is the pathway. The pair of electrodes are electrically connectable or connected so that the field providing the force can be defined therebetween along the length of the pathway.

[0038] In such an arrangement, the first manipulation assembly and / or second manipulation assembly may comprise at least one further electrode provided along the respective first pathway and second pathway which in part defines the individual controllable region. The control unit may be configured to operate the at least one further electrode to generate the first local force.

[0039] This may be one further electrode which is electrically connectable or connected to at least one of the pair of electrodes forming the first electrode set so that it forms a pair with at least one of these electrodes. The methods disclosed herein may accordingly comprise operating the at least one further electrode to generate the first local force. This defines an individually controllable region between the further electrode and at least one of the pair of electrodes, which electrodes together can be designated a pair of electrodes (e.g. a local electrode set). It will be noted that one of the pair of electrodes of the first electrode set (i.e. the pathway field-generating electrodes) can simultaneously be an electrode of the pair of the first electrode set and also of the local electrode set.

[0040] Migration along this particular individually controllable region may be controlled by the generation of a local force between the electrodes of the local electrode set. In this arrangement, the field can be generated across the entire pathway by the first electrode set and selectively a local field may further be generated within the individually controllable region. These can be simultaneously or separately generated.

[0041] Alternatively or additionally, there may a further pair of electrodes (i.e. there may be aplurality of further electrodes which are electrically connectable or connected to one another). That is, there may be an additional pair of electrodes (defining a local electrode set) to the pair of the first electrode set. This additional pair can be positioned so as to generate a local field therebetween across a sub -region of the respective pathway. Both electrodes may, in some cases, be located so as to form a local field wholly within the first pathway, and accordingly may be located between the pair of the first electrode set. The methods disclosed herein may accordingly comprise operating the pair of electrodes to generate the local force.

[0042] It will be appreciated that these arrangements can be combined. For example, there may be the further pair of electrodes defining the local electrode set but in addition at least one of these electrodes may be electrically connectable or connected to at least one of the pair of electrodes of the first set such that there are two individually controllable regions defined along the respective pathway: a first individually controllable region defined between the pair of electrodes of the local electrode set and a second individually controllable region between at least one of these electrodes of the local electrode set and at least one of the pair of electrodes of the first electrode set. There may be further combinations between the other electrodes of the sets. Indeed, it will be appreciated, that each of the electrodesforming part of the respective manipulation assembly may be electrically connectable or connected to each of the others and the control unit may be configured to operate these as pairs, as required.

[0043] The first electrode set can, instead, comprise a plurality of pairs of electrodes, each pair of electrodes being an electrically connectable or connected pair of field-generating electrodes. Each pair of electrodes of the first set can be provided along the pathway to define a region through the medium where there is a continuous field applied (i.e. the pathway). In other words, these is a continuous pathway defined by neighbouring and / or overlapping local fields produced by the plurality of pairs of electrodes. In this way, together the pairs of electrodes of the first electrode set define the respective pathway, but also define the individually controllable regions by providing the plural separate pairs of electrodes. The pairs of electrodes of the first electrode set can be operated together or individually, as required. It will be appreciated that, as with the embodiments discussed above, the various electrodes within the first electrode set can further electrically connectable or connected to other electrodes of the first electrode set (or indeed other electrodes), such that further forces can be provided between those additional pairings to create other regions. The control unit may be configured to operate these as pairs, as required.

[0044] The second manipulation assembly may be further operable to cause the at least a part of the sample (e.g. a component) to migrate from the first pathway along the second pathway. In other words, the second manipulation assembly may be configured or arranged so that the second pathway overlaps with the first pathway so that the sample or a component thereof can be transferred using the second manipulation assembly from the first pathway to the second pathway. In this way, the sample or a component thereof can be efficiently transferred without direct user intervention. For example, in existing separation systems, often there is a need for a user to manually retrieve a part of the sample or, in some cases, the entire track or channel along which the sample has moved, before it can be subsequently manipulated or analysed. By avoiding this, the risk of contamination or damage can be eliminated or reduced.

[0045] At least one of the first manipulation assembly and the second manipulation assembly may be operable to generate an electric field and / or a magnetic field to provide the respective force. In the methods disclosed herein, applying a force in the method may comprise applying an electric field and / or a magnetic field to the sample or a part thereof. The use of an electric field allows for movement of charged species along the respective pathway and is an efficient and effective means by which separation or manipulation can occur. This is also advantageous in that it can be combined with the other functionality disclosed herein. For example, using the medium or a modification element, the charge of a particular component (i.e. a species) can be modified at different points along the pathway, enabling different responses of the species to the applied electric field. This can in turn provide better separation. Similarly, the presence of an individually controllable region in which an electric field and / or magnetic field can be formed further enhance this selectivity and customisability. Where a magnetic field is used, this can be applied using an electromagnetic manipulation assembly configuredto apply a magnetic field across at least a portion of the respective pathway. The use of an electric field or magnetic field to provide the force is particularly advantageous with the structures and arrangements disclosed herein. For example, the application of an electric field or magnetic field can be achieved using electrodes, further allowing integration of this into a chip or integrated circuit structure and manufacture using traditional semiconductor manufacturing processes (e.g. CMOS manufacturing processes).

[0046] Where the force is applied by a field, variation of the force may be based on the strength or intensity (e.g. as measured by the input voltage (V / m or mV / m)) of the field applied, such as the electric field. The strength of the electric field may vary between the individually controllable region and at least another part of the field. It will be appreciated that in any of the embodiments mentioned here, although the resultant force of the electric field acting on a particular component will depend on a number of factors, including the charge on the component, the magnitude of the force will be determined by the magnitude of the electric field such that a higher V / m value will lead to a greater force acting on the species in the sample.

[0047] In addition to the profiles which can be formed across one pathway, it will be appreciated that different profiles can be formed across different pathways. For example, where each pathway is used for a separation, the first pathway may have a first force applied and the second pathway may have a second, different force applied. This can provide different degrees of separation, such as a gross or coarse separation and then a further separation of one or some of the components.

[0048] The system may further comprise a third manipulation assembly operable to cause at least a part of the component to migrate within medium provided at the sample surface along a third pathway, wherein the third manipulation assembly is arranged so that the third pathway is adjacent to or intersects the second pathway, wherein the control unit is further configured to control the third manipulation assembly so as to cause a part of the component within medium provided at the sample surface along the third pathway. The provision of a third manipulation assembly and a respective third pathway enables further manipulation of the sample or components thereof. For example, this can be used to provide a further degree of separation and / or may be used to manipulate the components already separated in the preceding pathway(s). This can then be used to transfer the components from the upstream pathways to a subsequent step (e.g. analysis, storage) without requiring a manual intervention from a user, thereby reducing risk of contamination or loss of sample. The system and method in this respect will therefore allow for up to a three-dimensional separation, including a one -dimensional separation with two manipulation steps or a two-dimensional separation with at least one manipulation step, depending on the requirements.

[0049] The method may therefore cause at least a part of the sample to migrate along a third pathway through the medium, wherein the third pathway is adjacent to or intersects the second pathway. Further, applying a force to at least a part of the sample may be so as to cause at least a part of the sample to migrate from the second pathway along the third pathway. Likewise, the control unit may be configuredto cause the third manipulation to cause at least a part of the sample to migrate from the second pathway along a third pathway.

[0050] In embodiments comprising a third pathway, the methods and systems may comprise a plurality of third pathways. The method may cause at least a part of the component provided to each second pathway to migrate out of the respective second pathway along a corresponding third pathway within the medium.

[0051] The third manipulation assembly may be operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along the third pathway so as to separate the sample into a plurality of components. In other words, the third manipulation assembly is arranged to separate the sample into a plurality of components. The method may comprise causing the component to migrate along the third pathway so as to cause separation of the component into a plurality of sub-components and / or a plurality of sub -components into further analytes. In this case, there can accordingly be at least two separations across the first, second and third pathways and, in some cases at least three. This can be particularly useful for complex samples and mixtures where components of the sample may comprise multiple sub -components and / or components may be closely spaced on one of the earlier pathways such that further separation is necessary.

[0052] The third manipulation assembly may be further operable to cause the part of the component to migrate from the second pathway along the third pathway. This capability allows for the sequential transfer of sample components between different stages of the separation process.

[0053] The third pathway may extend in the same or different direction to any of the preceding pathways. In some embodiments, the third pathway may extend in a plane different to the plane in which the first and / or second pathways extend. This allows for movement of the sample or a component thereof in more than one (and can be more than two) spatial dimensions further providing a means for extracting a particular component from the sample for further analysis. For example, a particular component could be moved in a plane or direction perpendicular to the plane in which the first and second pathways are formed so that it can be isolated from the remainder of the sample, for example to remove it from the medium and / or transfer for further analysis or storage. As set out above, the pathways may, but do not necessarily need to, extend in a linear direction. At least one of the first, second, and third pathways may extend at an angle relative to the sample surface, optionally wherein the first pathway extends in a first direction through the medium in a first plane, the second pathway extends in a second direction through medium in the first plane and at an angle relative to the first direction; and at least a part of the third pathway extends in a second plane intersecting and at an angle to the first plane. This configuration allows for three-dimensional manipulation of the sample components, providing a high degree of control over their movement and separation.

[0054] The method may comprise migrating the sub-component along the third pathway to an analysis location and determining at property of the sub-component at the analysis location.

[0055] It will be appreciated that there are also numerous other ways in which forces (beyond those resulting from the direct application of electric or magnetic fields to species affected by these fields) can be applied and separation can be affected, including for example:

[0056] (i) Applying a force may comprise providing a medium in which a property of the medium varies along a pathway so as to provide a force which acts on at least a part of the sample so as to causes the sample or a part thereof to migrate through the medium . This can define the corresponding pathway. In other words, the application of force is through the creation of an environment which leads to osmotic pressure or which causes diffusion across the pathway. This may be a property of the medium or it may be imparted on the medium by a corresponding manipulation assembly (where present in the method). For example, the methods (and systems) may comprise applying a force (which acts on at least one component) by providing a concentration variation or gradient within the medium along the pathway of at least one species. This can be a property of the medium or can provided by creating these regions through manipulations of species in the medium. For example, charged species (e.g. dopants not forming a part of the sample) within a medium could be manipulated by an electric field to create a particular concentration or charge gradient).

[0057] (ii) Applying a force may comprise applying a force to cause sedimentation of a sample (i.e. the separation of at least one non-soluble component from a liquid and / or for separating a plurality of non-soluble components from one another within a liquid). This may be application of a kinetic force to cause velocity sedimentation (measuring the speed at which a component settles) or isopycnic sedimentation (based on density of the components). The force acting on the particles may be gravity (i.e. the sample is provided at a first height in the medium such that gravity can move the samples to a plurality of different heights, thereby applying the force). The force may alternatively or additionally be a centrifugal or centripetal force device, such as applied by manipulation assembly (e.g. a centrifugal or centripetal force device (such as a zonal rotor)). One implementation of (ii) is separating cells.

[0058] (iii) Causing separation by a difference in at least one of adhesion, cohesion, and surface tension. This may be separating based on the difference in these features and interaction with the medium of the substrate surface or a feature provided on the substrate (such as a coating or functionalization). In some embodiments, these forces may also be used to apply the force so as to cause migration.

[0059] (iv) Applying a force may comprise applying a centrifugal or centripetal force to cause separation through the medium. This may be based on at least one of size of the components of the sample, shape of the components, density of the components, medium viscosity and rotor speed. For example, this can be used in combination with the separation of based on sedimentation (i.e. (ii)). At least one of (or a plurality or each) manipulation assembly may comprise a device configured to apply a centrifugal or centripetal force (such as a zonal rotor).

[0060] (v) applying a force may comprise applying a force using acoustic waves. For example, the respective manipulation assembly or assemblies may be configured to apply a force using sound-waves.These can also advantageously be used to apply a directional force on the sample or a component thereof. In one embodiment, the respective manipulation assembly or assemblies comprise an ultrasonic sound-wave generator, such as an ultrasonic transducer. By ultrasonic it is meant that the device generates high-frequency sound-waves which cause vibrations in a fluid, particularly in a liquid. For example, sound-waves with a frequency of greater than 18kHz, optionally 20-400 kHz and further optionally 40-80 kHz.

[0061] Combinations of any of the means for applying a force can be combined, including any of (i) to (v) with the use of an electrical field and / or magnetic field which acts directly on the sample. For example, the combination of (i) and an electric field can be used to move a sample or a part thereof through a gradient (which itself could be generated by a manipulation assembly) under the force of an electric field. Similarly, changing pH across a gradient (e.g. (i)) can be combined with an electric field to separate based on isoelectric point.

[0062] At least one of the first, second or third manipulation assemblies (or a plurality of, or each) may be configured to modify a property of the medium so as to provide a force which causes the sample or a part thereof to migrate through the medium. This may be in addition to alternatively to a field acting on the sample or a part thereof. For example, the second or third manipulation assemblies may modify a property of the medium along the pathway to create a gradient of the property across the pathway. The property (also referred to herein as “environmental property” or “medium property”) may be of the medium across the pathway or just the environment adjacent (i.e. next to or on) one part of the manipulation assembly (e.g. an electrode). By modification of property, it is meant that it to modifies a physical (e.g. material) or chemical property of the medium, the environment or the sample, such as by applying electrical energy or an electric field directly to the sample to change the property or by providing another form of energy (e.g. thermal energy). In some embodiments, this may be a physical or chemical property of the medium. The property can include physical properties, such as thermal properties (e.g. temperature) and / or viscosity, and / or chemical properties selected from at least one of the pH, electrical conductivity, thermal conductivity and / or ionic strength of the sample or a part thereof. The property may be modified directly, e.g. by direct application of heat to change the temperature, or indirectly for example by causing a chemical change which in turn changes the property, such as hydrolysis or electrolysis due to application of electrical energy which can change the pH. For example, the manipulation elements can be configured to create an electrical field which can cause a localised change in a part of the medium.

[0063] Accordingly, at least one of (or a plurality of, or each) the first, second or third manipulation assemblies is configured to modify at least one of the concentration of at least one component of the medium

[0064] For example, certain molecules are only charged within small pH ranges and may not bind to the corresponding moiety if not charged. Modification of the pH can change the propensity of the capture species and target analyte to bind, which can provide additional information on binding andkinetics of binding. In some cases, this can lead to dissociation. Changes in pH and / or ionic strength can also be used where the sample comprises other non-analyte species, by modifying the solution to a point where e.g. non-selective binding is reduced or eliminated. Moreover, pH is a critical factor in many aspects of biological processes and, indeed, is heavily susceptible to variation in biological samples, for example due to biological processes (e.g. CO2 or lactate production by cells). The ability to control this in situ and without requiring buffers or acids / bases provides a significant improvement.

[0065] The system (or in embodiments, apparatus) may further be configured such that at least one of the manipulation assemblies or, further, a separate manipulation assembly (which can have the configurations of or properties of any of the other assemblies disclosed herein) may be operable to cause a sample moving along any of the pathways to additionally move towards the sample surface, a sensing element (where present) and / or a modification element (where present). In other words, it is operable to apply a force which causes a sample or a component thereof to move towards a desired region of the system. Often sensing of components within a system (e.g. where no force is applied) requires diffusion of the targets to a sensor. For example, where the sample has been separated along a path and there are sensing elements configured to detect these, the force causing migration may no longer be applied and so sensing may be limited by diffusion of the component to the sensing element (e.g. on the sensing surface). A local or general force may be applied to drive the component towards the surface and increase the speed of interaction. This force is different to the individually controllable region. Similarly, the method may comprise applying a field (e.g. an electric field) to the medium so as to cause movement of a part of the sample towards the sample surface, a sensing element (where present) and / or a modification element (where present). Such a method can speed up measurement time by causing movement of the species in the sample to migrate to a particular region at a movement speed which is greater than diffusion. This can reduce the time of or reduce the need for an incubation period. This may be considered to be separate to movement along the pathways, since there can be expected to be some of the part of sample already around the destination in question; rather, this seeks to speed up this process.

[0066] The system (or in embodiments, apparatus) may further be configured such that at least one of the manipulation assemblies or, further, a separate manipulation assembly (which can have the configurations of or properties of any of the other assemblies disclosed herein) may be operable to collect (e.g. concentrate) at least one component or sub -component in a region of the medium. In other words, a diffusion region of the at least one component or sub -component may be collected into a smaller region or spot. This can help to avoid issues with low concentrations of components / sub- components within a sample and to counter drift caused by diffusion of the sample. For example, this may be achieved using at least one collection electrode located on the region where this is to be collected and at least one (but optionally a plurality of electrodes) around the collection electrode and using these to generate an electric field which moves the component / sub -component to the collection electrode.

[0067] The system (or in embodiments, apparatus) may be configured such that the first manipulation assembly is operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway so as to separate the sample into a plurality of components. For example, the methods may comprise causing the sample to migrate along the first pathway so as to cause separation of the sample into a plurality of components, wherein causing a part of the sample to migrate along the second pathway comprises causing at least one component to migrate along the second pathway. Further, causing a part of the sample to migrate along the second pathway may cause separation of the component into a plurality of sub -components. This separates the sample into distinct components based on their respective properties along the first pathway which components can then be manipulated or moved into the second pathway.

[0068] For example, the second manipulation assembly may be operable to cause a plurality of components to migrate from the first pathway, each component of the plurality migrating along a separate second pathway adjacent to or intersecting the first pathway. In other words, there is a plurality of second pathways. This allows for the simultaneous manipulation of multiple components, increasing the throughput and efficiency of the separation process. In this case, the second manipulation assembly may have individually actuatable sub-assemblies for each or plural of the second pathways or there may be one which controls migration along each. Or there could be a plurality of second manipulation assemblies, each one associated with a separate second pathway. The method may comprise causing a plurality of components to migrate from the first pathway, each component of the plurality migrating along a separate second pathway adjacent to or intersecting the first pathway. Each second pathway is adjacent to or intersects a different point along the first pathway. This advantageously allows for interrogation of and interaction with a plurality of components of the sample. This can be used to determine multiple constituents and may be used to identify a signature or fingerprint of the overall sample.

[0069] It will be appreciated that, alternatively, movement along the first pathway may be used to position the sample for subsequent separation along the second pathway (and optionally further pathways). Accordingly, in such a case or in cases where there is separation along both the first and second pathways, the system (or in embodiments, apparatus) may be configured such that the second manipulation assembly is operable to apply a force to a sample to cause the sample to migrate within the medium along the second pathway so as to separate the sample into a plurality of components or separate at least one component into a plurality of sub -components.

[0070] In addition to the force, the system may be provided with a separation component provided on the sample substrate which interacts with at least a part of the sample as it is separated. This may be a component provided within the medium (e.g. particles) or provided on the sample surface (such as a coating or functionalization). Examples include antibodies provided on the sample surface, beads (which can be functionalized) or surface profiling on the sample surface.Combined use

[0071] The electrodes in the system and provided along or interacting with the pathway can further be used to interrogate the sample or the components thereof. For example, an electrode or electrodes may be used to obtain a signal relating to a property of the sample or components thereof, including electrical proprieties, such as current, voltage, resistance, capacitance, impedance or conductance.

[0072] For example, the electrode set(s) and / or the at least one further electrode may be further operable to provide a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway. Where there are plural electrodes, such as the electrodes of the local electrode set, each may be further operable to provide a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway. The control unit may be configured to address the electrode set(s) and / or the at least one further electrode to obtain the signal and may, further, be configured to determine a property of the component based on the signal. For example, the property may be an intrinsic property of the product, such as the structure, melting point, etc. In particular, this may be analysis of the structure of the product, for example. In another embodiment, the property may be an extrinsic (or relational) property. In some embodiments, the property may be a concentration of the product on a capture site defined by a capture species, or on the sample surface. For example, this may be determination of a number of the individual product molecules or entities on the capture site. The methods disclosed herein may include obtaining a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway using the at least one further electrode or the electrode set(s). The position along the first pathway is a point at which the first pathway and the second pathway intersect or overlap. This advantageously utilises the electrodes used for manipulation of the sample to further interrogate the sample, providing additional functionality without increasing the number of components which need to be provided. This can also speed up and improve the robustness of the analysis, since the data is provided in situ, rather than using a separate device and requires no transfer.

[0073] The control unit may be further configured to determine a property of the component based on the signal provided by the electrode set(s) and / or the at least one further electrode, optionally wherein the control unit is further configured to control the first manipulation assembly and / or the second manipulation assembly based on the determination of a property of the component of the sample. The methods disclosed herein may further comprise determining a property of the component based on the signal provided by the at least one electrode, optionally further comprising controlling the step of applying the force to the sample or component along the first pathway and / or the second pathway based on the determination of a property. This allows for manipulation of the sample components based on sensed data, rather than simply determining the outcome of a manipulation and separation event. This can lead to more accurate and efficient separation and analysis. For example, the forces applied or the operation of the individually controllable region(s) may be dependent on the detection of a component, such as in a particular region, or an amount. For example, the electrode set(s) and / or the at least onefurther electrode may provide a signal indicative of a property of a component of the sample at a particular position along the respective first pathway and / or second pathway. This can be useful where the sample is provided on one of these electrodes to begin the manipulation or separation or where a component or matrix reaching the other electrode may indicate that manipulation or separation is complete.Modification element

[0074] The system may further comprise at least one modification element provided along the first and / or second pathway operable to: (i) interact with at least a part of the sample migrating along the respective first and / or second pathway and / or (ii) influence the migration of the at least a part of the sample along the respective first and / or second pathway. The control unit may be configured to operate the modification element so as to cause the modification element to interact with and / or influence the migration of at least a part of the sample within (e.g. migrating along) the respective first and / or second pathway. The method may also comprise providing at least one modification element along the first and / or second pathway, wherein the at least one modification element is operable to interact with at least a part of the sample migrating along the respective first and / or second pathway and / or influence the migration of the at least a part of the sample along the respective first and / or second pathway. Influencing the migration may be caused by a direct interaction with the sample, for example by creating an attractive force or slowing which slows down migration or by retaining the sample on the modification element, or it may be achieved by modifying the environment of the medium, which in turn may change how the sample or a part thereof moves through the medium. The former may accordingly be an interaction directly with the sample and the second may be indirect and, hence, just influence the migration by affecting another part of the device. These modifications can have significant effects on the migration patterns of the sample components, allowing for more effective separation based on their physical and chemical properties. For example, it can modify how at least one component of the sample interacts with the medium or the force applied. For example, a change in pH may cause a change in charge of at least one of the components, and in turn this may modify how an electrical field interacts with the component, changing the way it migrates through the medium. Interaction may, in some cases, comprise influencing migration but in others may have no influence on migration but cause other effects. For example, additionally or alternatively, it can also modify the interactions between the various components of the systems, such as electrodes, with the samples. This can change whether or not an electrode (e.g. acting as a sensing element) interacts with the electrode, for example increasing or decreasing the response to the electrode or the interaction of a component with a capture agent provided on a sensing element.

[0075] The modification element may be used to locally modify a property. That is, it may be configured to modify a property (also referred to herein as “environmental property” or “medium property”) of the medium or environment adjacent (i.e. next to or on) the modification element or onlymodify the sample within a region local to the modification element. By “property” and modification of property, the same definitions and possibilities are present as for the first, second or third manipulation assemblies. This can be used to create localised regions having a different property. Examples include the modification unit being configured to create an electrical field which can cause a localised change in a part of the medium. Another example is a temperature increase which can decrease incubation time with a sensing element (or a capture species provided thereon) by increasing the rate of binding (Kon). In the case of some biomolecules, such as antibodies and a corresponding antigen, without wishing to be bound by theory, the rate of binding at or close to 37 °C is higher than that at room temperature (e.g. 21 °C), even if limited to the region adjacent the sensing surface only (i.e. without increasing the bulk temperature). Ionic strength and pH (which are linked) also modify the binding parameters. For example, certain molecules are only charged within small pH ranges and may not bind to the corresponding moiety if not charged. Modification of the pH or ionic strength can change the propensity of the capture species and target analyte to bind, which can provide additional information on binding and kinetics of binding. In some cases, this can lead to dissociation. Changes in pH and / or ionic strength can also be used where the sample comprises other non-analyte species, by modifying the solution to a point where e.g. non-selective binding is reduced or eliminated. Moreover, pH is a critical factor in many aspects of biological processes and, indeed, is heavily susceptible to variation in biological samples, for example due to biological processes (e.g. CO2 or lactate production by cells). The ability to control this in situ and without requiring buffers or acids / bases provides a significant improvement.

[0076] The modification element may comprise or be a thermal device, such as a heater or a cooler. This can be used for the purposes set out above or may be used to generally increase the temperature of the environment and / or medium. This can be locally (i.e. configured or operable to modify the temperature of the medium and / or a part of the sample adjacent the thermal device) or globally across the entire sample surface / within the medium. For example, by raising the temperature (e.g. by at least 1 °C, or at least 5 °C), the entire process may be sped up, by increasing mobility, reaction times and / or binding. For example, the modification unit can be used to provide heating for e.g. PCR, isothermal amplification, association and dissociation of components with the modification element or a capture site (such as components / sub-components with a corresponding capture probe), modifying components (such as denaturing, annealing and elongation). The control unit may be configured to operate the thermal device to heat or cool the medium and / or a part of the sample. The methods disclosed herein may comprise operating the thermal device to heat or cool the medium and / or a part of the sample. The modification element may be provided with a functional component on the surface, such as a capture agent or species (discussed below) or another functional element (such as a component which causes a chemical or biological reaction in conjunction with the modification - examples include an enzyme, such as for digesting proteins into peptides).

[0077] The modification unit or modification element used in the system and methods may comprise an electrode (e.g. a manipulation electrode) or a plurality of electrodes operable to interact with at least a part of the sample. An electrode can be used to provide a chemical, thermal, or mechanical manipulation, depending on the nature of the sample and the specific requirements of the separation process. For example, it can be used to can alter properties such as pH, electrical conductivity, or temperature of the medium, thereby affecting the behaviour of the components and facilitating their separation and analysis. For example, these property modifications can have significant effects on the migration of the components, allowing for more effective separation based on their physical and chemical properties. Where there are a plurality of manipulation electrodes provided along a pathway, these may be operable to act together. It will be appreciated that the electrodes forming a part of the manipulation assemblies (e.g. the at least one further electrode or local pair of electrodes) may act as the modification element under the control of the control unit (i.e. where the control unit is configured to operate them as such). As such, the features set out herein above, in respect of the functionality of the modification element may equally apply to the electrodes forming a part of the manipulation assemblies, if used in this manner (e.g. if controlled by a property modification unit and / or the control unit).

[0078] The electrode may be provided with a surface comprising a capture agent configured to bind to at least one component of the sample such that the component can be retained on the electrode. This feature allows for the isolation and subsequent analysis of a components at a point along the pathway. The method may further comprise operating the modification element so as to retain at least one component of the sample thereon.

[0079] The modification element may alternatively or additionally be configured to influence migration through the use of a trapping field configured to trap a sample or component thereof in a particular region, for example a region along the pathway. This can be an electrical, magnetic or optical trapping field.

[0080] The modification element may be provided on the substrate and located along a pathway (e.g. the first pathway and / or second pathway). This further allows for integration of this into a chip or integrated circuit. Further, providing this on or adjacent the sensing surface enables the modification of at least a part of the sample in proximity to the sensing surface. Specifically, the environmental property / properties can be modified in situ and in the part of the sample (localised region) where the measurement is occurring. This can speed up measurement time, for example by avoiding separate pretreatment and modifying the bulk sample, and enables the internal modification of particular properties without needing additional reagents (e.g. by creating property gradients within the sample). Moreover, this enables the real-time monitoring of any impact resulting from the modification, thereby providing additional information regarding the nature of the sample and the target analyte. For example, response to the change of environmental property, for example rates of change in measurement signal, can be monitored.

[0081] The system may comprise a property modification unit comprising the modification element, where the property modification unit is configured to modify a property of medium provided at the sample surface using the modification element so as to influence the migration of at least a part of the sample migrating along the respective first and / or second pathway. The property modification unit may comprise a control unit or controller (which may be the control unit of the system or a separate control unit or controller) configured to operate the property modification unit, for example. The method may further involve modifying a property of the medium so as to impart variation in the medium property along at least a part of the first pathway and / or the second pathway. As such, the property modification unit may be configured to modify (and the method comprising modifying) at least one of pH, electrical conductivity, a thermal property (e.g. temperature of), and / or ionic strength of the medium (e.g. by operating the modification element).

[0082] The property modification unit may be configured to form a first property region across a first portion of one of the first pathway and / or second pathway and form a second property region across a second portion of the respective pathway such that the property varies across the respective pathway. Similarly, the method may comprise modifying a property may include forming a first property region across a first portion of one of the first pathway and / or second pathway, and forming a second property region across a second portion of the respective pathway such that the property applied varies across the respective pathway. The method may also comprise providing the property modification unit set out herein to provide the property regions. This gradient of properties can be used to create zones of different conditions along the pathways, which can be used to separate components based on their responses to these conditions. This can be achieved, for example, by having a first modification element in or adjacent the first portion and a second modification element in or adjacent the second portion and operating them accordingly. The property modification unit may comprise a first modification element set (e.g. comprising at least one modification element) operable to modify a property of the medium in the first portion and a second modification element set (e.g. comprising at least one modification element) operable to modify a property of the medium in the second portion. These may be electrode sets comprising at least one modification element in the form of an electrode.

[0083] The modification element may be operable (or operated) to cause a transformation or conversion of the sample or a component thereof. In the methods and systems, a sample or a part thereof can be transformed or converted into at least one species or product. By “conversion”, this refers broadly to any process that transforms a component or group of components from one form to another, including but not limited to: (i) a chemical reaction wherein one compound is converted into another, for example via synthesis, rearrangement, polymerization, isomerization, or substitution; (ii) a destructive transformation wherein a species is broken down or separated into component parts, for instance by decomposition, fragmentation, or pyrolysis; or (iii) any other process facilitating a change in composition or structure. This may include single -step or multi-step conversions (or “transformations”). “Conversion” may therefore encompass a variety of mechanisms, for exampleinstigated by the ’’stimulus” applied by a modification element, which is described in more detail, below. A stimulus (such as heat or cooling, voltage, current) may therefore be applied to the sample or a component thereof to cause the transformation. This could be used to release components from the sample (for example, to cause cell lysis where the sample is a cell), break components down into subcomponents (for example, causing molecules or other components to degrade into by-products) or similar. Examples include converting proteins to peptides, nucleic acids to bases and polymers to monomers. This adds an additional means of interrogating the sample and may provide further selectivity or confirmation of the identity of a component where it breaks into identifiable by-products. This is readily achievable using structures of systems disclosed herein since a transmembrane breakdown potential for a cell is typically around IV such that an electrical field strength of ~0.3 x 105V / m can be used for electrical lysis of cell with a 20pm diameter.Sensing element

[0084] The system may further comprise a sensing element operable to provide a signal indicative of a property of a component of the sample at a position along at least one of the first pathway and the second pathway. The sensing element can accordingly be used to monitor the separation and manipulation properties of the sample. This can also further integrate analysis which may otherwise be carried out separately into the process, speeding up the overall measurement(s) and analysis. The sensing element may be an optical sensing element, thermal sensing element, pH sensing element, conductivity sensing element, fluorescence sensing element or a sensing element configured to detect a chemical property. Electrochemical sensors such as amperometric, potentiometric or conductometric sensors may also be employed. The sensing element may be located along the pathway, for example on or in the substrate. Of course, where there are further pathways, at least one sensing element may further be operable to provide a signal indicative of a property of a component of the sample at a position along the further pathway (s).

[0085] The control unit may be further configured to determine a property of the component based on the signal provided by the sensing element. The control unit may be further configured to control the first manipulation assembly and / or the second manipulation assembly based on the determination of a property of the component of the sample. This feedback can be used to optimise the manipulation and separation of the sample.

[0086] The sensing element (or “sensing device”) provides the measurement signal and therefore may be addressable to provide the measurement signal. For example, the sensing element may comprise an electrode, which electrode may be addressable to provide the measurement signal. In some embodiments, there may be plural electrodes. Where there are plural electrodes, each may be individually addressable. A sensing layer (e.g. a dielectric layer) may also be provided on or adjacent the electrode to enhance or enable the response of the electrode to a particular component or analyte. In other embodiments, the sensing element may comprise a layer or structure through which current is passed (for example, where the sensing element comprises is a resistive layer or structure locatedbetween first and second electrodes). In some embodiments, a portion of the substrate defining the sample surface and / or a portion of the medium may define the sensing layer or structure between electrodes and the sensing element may comprise a pair of electrodes arranged to interrogate the portion of substrate and / or medium such that sample or a component in this particular region can be interrogated.

[0087] Where the sensing element is or comprises an electrode, the electrode can be a separate component to the various electrodes disclosed herein (e.g. the electrodes of the sets forming the manipulation assemblies, including the further electrode(s)) or the sensing element can be one of these electrodes. Accordingly, it will be appreciated that the electrodes forming a part of the manipulation assemblies (e.g. the at least one further electrode) may act as the sensing element. As such, the features set out herein above, in respect of the functionality of the sensing element may equally apply to the electrodes forming a part of the manipulation assemblies, if used in this manner (e.g. if controlled by a property modification unit and / or the control unit).

[0088] Where the sensing element is or comprises an electrode, the electrode may comprise a capture agent provided thereon configured to retain a component of the sample on the electrode. A capture agent may be any composition or element configured to hold and retain a component of the sample. For example, immobilization of a target species (e.g. the component) can occur through e.g. covalent-like interactions (e.g., chemisorption of anchor species onto the surface through chemical bond formation) and non-covalent-like interactions (e.g., physisorption of capture species onto the surface through weaker, often van der Waals, interactions) depending on the identity of the capture agent and the component. This capture agent may be selective to a particular component or type of component. It may be a capture species (i.e. a molecular species or functional group) configured to selectively bind with a component (e.g. a target species) which can be present in the sample . The capture species may comprise a protein (e.g. an enzyme), a peptide, a carbohydrate, a nucleic acid, an aptamer or a combination thereof. This allows for various uses. For example, the protein may be or comprise an enzyme or an antibody. Some examples include a single or double stranded DNA chain. An aptamer may be defined as an oligonucleotide or peptide configured to bind the analyte. Such an aptamer may, for example, be configured to interact with, for example bind, various analyte types, such as small molecules, for example amino acids or amines, proteins, metal ions, and microorganisms. Other capture agents may include e.g. molecular sieves (such as gels) for physical capture. Other examples of immobilization of capture agents include surface functionalization techniques such as thiol-gold or silanization (GOPS, APTES) or Poly-L-Lysine (Electrostatic). Capture species may be plural, such that in some cases, there may be a plurality of capture species (i.e. plural of the same capture species) on each sensing element.

[0089] The method may further comprise obtaining a signal indicative of a property of a component of the sample at a position along at least one of the first pathway and the second pathway. The method may further comprise determining a property of the component based on the signal obtained, optionallyfurther still the method may comprise controlling the step of applying the force to the sample or component along the first pathway and / or the second pathway based on the determination of a property.

[0090] The methods may comprise determining a concentration and / or a copy number of a component. The system may be configured to determine a concentration and / or a copy number of a component. This may be achieved by providing a plurality of different concentrations (e.g. dilutions) of at least a part of the sample, with each concentration being provided to a separate pathway (for example from other pathways); moving or causing the components to each move along their respective pathway to a sensing point (e.g. a sensing element) and determining the concentration and / or copy number.Substrate and Medium

[0091] The system and method comprise a substrate defining the sample surface. The substrate may be a planar surface or a non-planar surface. For example, the substrate may comprise multiple substrate portions, where the substrate portions are in different planes. For example, these may be angled with respect to each other or in opposing relationship (such that medium is provided therebetween). Each substrate portion may be planar. Where present, the substrate portions may abut one another or may be separated by joined by a liquid or gel medium.

[0092] The substrate may be formed from or by a layer or, where there are plural substrate portions, may be formed of or by a plurality of layers or sections forming the surface, for example.

[0093] Medium may be provided onto or over the sample surface but may further extend into the substrate, in some embodiments, and / or beyond the sample surface. The substrate may be formed of or comprise any suitable material. For example, it may be a polymer layer, a glass layer, a glass-ceramic layer, a ceramic layer, a metal oxide layer, a metal nitride layer, a silicon -containing layer (e.g. silicon, silicon dioxide, or silicon nitride), a gallium-containing layer (e.g. gallium nitride or gallium arsenide) or combinations thereof.

[0094] The substrate may comprise at least one opening provided at the sample surface, wherein at least one of the pathways (or a part thereof) extends into the opening. As such, the method may further comprise providing a substrate defining the sample surface, the substrate comprising an opening provided therein at the sample surface, wherein at least one of the second pathway and the third pathway is arranged at an angle relative to the sample surface adjacent or at the opening (e.g. the portion surrounding the opening) and extends into the opening so that migration of the component or the part of the component, respectively, causes movement of the component or the part of the component through the opening. Where there is a third pathway, one of the second pathway or the third pathway may extend into the opening. That is, the pathway extending into the opening may be arranged at an angle relative to the sample surface and extend into the opening so that migration of the component or the part of the component, respectively, causes migration through the opening. A well, channel or through hole may accordingly form a part of the pathway or define a part of the pathway and define the opening. Where a closed recess is formed (e.g. a well or channel), the nadir or base of the recess maydefine the end of the pathway. There may be a plurality of openings, with a plurality of pathways extending into a respective opening. The openings may be openings of channels, wells or through holes formed in the substrate and optionally further layers provided below or adjacent the substrate. Accordingly, the substrate may comprise at least one channel, well or through hole. The presence of openings in the substrate or a portion thereof provides significant additional functionality. The openings enable transfer of components of the sample into an enclosed or separate region, where further analysis or storage can occur. For example, components or specific analytes of the sample could be collected in wells or channels formed in the substrate so as to concentrate them. This can be useful where there are numerous samples or aliquots of samples from which the same component can be extracted from each and then recombined in a well or device positioned below a through hole.

[0095] Openings through a substrate, such as a well, channel or through hole, have been found to be particularly useful for sensing. Specifically, the system may further comprise sensing a property of a component of the sample within the well, channel or through hole. For example, a sensing element (as set out above) may be formed next to, around or within the well, channel or through hole. For example, there may be an electrode at least partially surrounding this or a pair of electrodes positions either side of the well, channel or through hole. Sensing elements using electrodes in this way have been found to provide a particularly useful tool for determining a property of a component, since the relative position of the analyte within or around the structures can have a significant effect on the properties and, hence, the signal from the sensing element. The measurement signal can be indicative of an impedimetric property (e.g. dielectric property (e.g. permittivity), resistance, capacitance, impedance, conductance, or a combination thereof) of the substrate and a component received between or within the well, channel or through hole modifies the impedimetric property. That is, it measures an impedimetric property, which in one embodiment is based on the permittivity of the sensing layer. This can function by the sensing layer having a first impedimetric property (e.g. permittivity) when in the presence of no sample (defined by the sensing layer and any fluid (e.g. a buffer or air) between the structures), and then receipt of the analyte between or within the structures can have significant impact on the impedimetric property (e.g. permittivity). Through holes are advantageous as these do not have a truncated structure and therefore the signal can be monitored as it passes through the through hole. Moreover, this can advantageously be used to provide an outlet for removal of components therethrough. Control of the interaction of samples or components thereof with the openings can lead to generation of a sample or component-specific signal. Each sample or component will interact differently with the surface and / or opening such that continuous measurement during the interaction and / or differential measurement can lead to a fingerprint signal specific to that sample or component.

[0096] The openings may have a maximum diameter of from 1 nm to 500 pm. For sensing individual species (such as molecules), this may be from Inm to 2 pm such as from 1 nm to 1000 nm, or 10 nm to 100 nm. A through hole, well or channel defining the opening may also have a maximum diameter of from 1 nm to 2 pm, such as from 1 nm to 1000 nm, or 10 nm to 100 nm. The diameter of the openingmay be the same as the corresponding through hole, channel or well. In other cases (including sensing) the dimensions may be larger, such as from 0.5 pm to 500 pm, or 5 pm to 200 pm, for example where the sample comprises cells.

[0097] In some embodiments, the depth of the well, channel or through hole may be less than or equal to 10 pm, for example less than or equal to 5 pm, for example less than or equal to 2pm, less than or equal to 800 nm, or less than or equal to 500 nm. In one embodiment, the wells, channels or through holes have a depth of from 10 nm to 10 pm. This may be 10 nm to 5 pm, such as 10 nm to 2 pm, 10 nm to 1000, 10 nm to 500 nm, 50 nm to 1000 nm, 50 nm to 800, 50 nm to 500 nm, 100 nm to 1000 nm, 100 nm to 800, or 100 nm to 500 nm. Exemplary combinations include a maximum diameter of 1 nm to 2 pm and a depth of 10 nm to 10 pm, such as 100 nm to 1000 nm maximum diameter and 10 nm to 2 pm depth. In one embodiment, the depth of the recesses or through holes is from 0.15 pm to 1 pm. The through holes or opening may be provided with or comprise a mesh or nanoporous structure, which can act as a further interrogator of the sample and / or a fdter where the through hole can be used to further separate the sample.

[0098] The system and method may further comprise a medium provided at (e.g. on) the sample surface. The medium may be or comprise any matter through which a sample can move. This may be a fluid medium, such as a gas (e.g. air), liquid or gel medium. Example gels include silica, alumina, cellulose, agarose or polyacrylamide gels. For example, an immoniline drystrip gel (“IPG gel”), an isoelectric focus gel (“IEF gel”) and a polyacrylamide gel (polyacrylamide electrophoresis (“PAGE”)). Other examples of media include size exclusion gels, affinity (modified) matrices, activated charcoal, native gels, and sample -dependent media. A liquid medium may be a liquid buffers or solvents. The medium may contain additives in some cases to enhance separation, such as detergents, denaturants, or specific binding molecules. In certain aspects, the medium may have a property gradient, such as a pH or ionic strength gradient, to facilitate separation based on properties such as isoelectric points or charge. It will be appreciated that the medium may be provided together with the sample.

[0099] The medium may advantageously comprise different regions. For example, the medium may comprise a first medium portion having a first medium property and through which at least a portion of the first pathway extends; and a second medium portion having a second medium property and through which at least a portion of the second pathway extends, wherein the first and second medium properties are different. By different it is meant that the medium regions have a different properties which have different interactions with a component or sample. For example, the medium may have regions with different physical states (e.g. at least one gel region and at least one liquid region or regions having different viscosities (whether gel or liquid)) or different compositions (base composition of the medium and / or different concentrations of specific functional components). This configuration allows for the creation of zones with different properties along the pathways, which can be used to separate components based on their responses to these properties. The medium regions may correspond to various other components disclosed herein. For example, the second medium region may correspond toan individually controllable regions. Alternatively or additionally, a different medium region may be provided above a sensing element and / or one of the openings in the substrate, where present. In another example, this may include particles within the medium, such as beads, which influence migration and enhance separation. In another example, there may be nano or microstructured surfaces (e.g. this may be the sample surface) to interact with the sample and influence migration.

[0100] The medium may comprise additives and / or the substrate or surface of the medium may be functionalised, for example to provide further functionality. For example, the medium and / or surface can be provided with a target specific species, such as a species configured to bind to a particular target. This can be a label, such as a fluorescent label. Examples include SYBR Green dye and ethidium bromide). The medium and the sample may be immiscible. Where this is the case, the sample (and components thereof) may be manipulated using an electrowetting technique. Where the sample is aqueous or provided within an aqueous matrix, the substrate may be hydrophobic such that the aqueous part can be manipulated through the medium on the surface. Such methods and systems may further use a modification element to control the immiscible sample, such as modifying temperature and / or pH.Electrodes

[0101] Numerous embodiments of the systems and method disclosed herein employ electrodes of various types, including for generating fields and as modification elements. These electrodes may each be formed from or comprise gold, silver, copper, platinum, nickel, titanium, titanium nitride, ruthenium, a conducting polymer, a conductive carbon -based material (e.g. graphene) or combinations thereof. It will be appreciated that where an electrode is used, this may be a part of any suitable electrical circuit or system. For example, the electrode may be a working electrode and there may be a counter electrode provided in electrical connection with the working element, and there may further be a reference electrode (or pseudo reference electrode) provided as part of this electrical system. Where there are pairs of electrodes, these can act as working and counter electrodes.System Architecture and Control Unit

[0102] The systems comprised herein each comprise at least a substrate, a first manipulation and a second manipulation assembly. These components may be provided separately or may be provided as a single apparatus. Accordingly, there may be an apparatus comprising the substrate, the first manipulation assembly and the second manipulation assembly. Where there are other components such as a third manipulation assembly, a manipulation element, a property modification unit, and similar, these too may be a part of the apparatus. Provision of these components as disclosed herein as an apparatus advantageously lends itself to semiconductor manufacturing techniques and miniaturisation.

[0103] The system comprises a processing unit or control unit. The control unit may be or comprise a processor or controller. The control unit may be implemented in any suitable manner, with software and / or hardware, to perform the various functions required. One or all of the units may, for example, employ one or more microprocessors programmed using software (for example, microcode) to performthe required functions. It will be understood that the control unit may be or run on a single controller or processor or may be distributed over several computers and locations (e.g. connected via internet), such as a cloud -based computing infrastructure. Examples of processor components that may be employed in various embodiments include, but are not limited to, conventional microprocessors (e.g. a central processing unit (CPU), a digital signal processor (DSP)), application specific integrated circuits (ASICs), and field-programmable gate arrays (FPGAs). The control unit may include, but is not limited to, PCs, workstations, laptops, PDAs, palm devices, servers, storages, and the like. Generally, in terms of hardware architecture, the control unit may include one or more processors, memory and one or more I / O devices that are communicatively coupled via an interface. In various implementations, the control unit may be associated with one or more non-transitory storage media such as volatile and non-volatile computer memory including any one or combination of volatile memory elements (e.g., random access memory (RAM), such as dynamic random access memory (DRAM), static random access memory (SRAM), etc.) and non-volatile memory elements (e.g., ROM, erasable programmable read only memory (EPROM), electronically erasable programmable read only memory (EEPROM), programmable read only memory (PROM), tape, compact disc read only memory (CD-ROM), disk, diskette, cartridge, cassette or the like, etc.). The memory can have a distributed architecture, where various components are situated remote from one another, but can be accessed by the control unit. The non-transitory storage media may be encoded with one or more programs that, when executed on one or more processors and / or controllers, perform the required functions. Various storage media may be fixed within a processor or controller or may be transportable, such that the one or more programs stored thereon can be loaded into the control unit.

[0104] In some non-limiting examples, the system includes a user interface, such as a display. Alternatively or additionally, the system may include a communications interface device, such as a wireless transmitter, configured to transmit data, such as the property determined by the property determination unit, to an external device, such as a personal computer, tablet, smartphone, remote server, etc.

[0105] Data between parts of the system (such as processors) or with external components may be via communications interfaces. The interfaces may communicate via a physical connection / transceiver.

[0106] Additionally, or alternatively, the communication interfaces may each comprise a wireless transmitter. Put another way, the communication interface and host controller may communicate via a non-physical connection. For example, this may be via a radio, RFID, NFC, Bluetooth, or Wi-Fi connection. Additionally, the transfer of data may be bi-directional, in that the wireless transmitted may also receive data from the host controller (e.g., the instruction as described above). Data transmitted between the system component and / or with external devices may be encrypted. The control unit may be configured to process the signals or data to generate an encrypted signal. The communication interface may then be configured to transmit the encrypted signal.

[0107] The control unit may be configured to analyse the signals and / or data using a machine learning algorithm trained to receive this data and output processed data based on the received data. For example, it may reive a measurement signal or the data derived therefrom. That is, the control unit provides the signal and / or data to a machine learning algorithm. The machine learning algorithm may be trained to process the signal to generate a value indicative of the property of the component being analysed.

[0108] The analysis of the signals and / or data may include a pattern recognition module configured to determine whether the signal and / or data (which may be processed data) corresponds to a predetermined or library signal or data. Accordingly, a signal and / or data can be matched with a known signal and / or data to identify a particular sample, component and / or sub-component.

[0109] Methods of training a machine -learning algorithm are well known. Typically, such methods comprise obtaining a training dataset, comprising training input data entries and corresponding training output data entries. An initialized machine -learning algorithm is applied to each input data entry to generate predicted output data entries. An error between the predicted output data entries and corresponding training output data entries is used to modify the machine -learning algorithm. This process can be repeated until the error converges, and the predicted output data entries are sufficiently similar (e.g. ±1%) to the training output data entries. This is commonly known as a supervised learning technique. Accordingly, the machine learning algorithm may be trained using a set of measurement signals and / or data. This training data may be specific to the type of component such that the machine learning algorithm is tuned to produce component data based on measurement signals and data for a specific sample, component or sub-component. Alternatively, the machine learning algorithm may be trained using training data for a variety of different samples, components or sub-component, such that the machine learning algorithm is capable of producing data for a variety of components. In one aspect, there is provided a computer program comprising computer program code which is configured, when said computer program is run on one or more physical computing devices, to cause said one or more physical computing devices to implement the methods disclosed herein.

[0110] In one aspect, there is provided one or more non -transitory computer readable media having a computer program stored thereon, the computer program comprising computer program code which is configured, when said computer program is run on one or more physical computing devices, to cause said one or more physical computing devices to implement the methods disclosed herein.

[0111] In one embodiment, the system may further comprise a signal processing unit configured to process signals received relating to measurements, for example from the sensing elements and electrodes set out herein. A property determination unit may receive processed signals and determined the property based on the processed signals. The property determination unit may, in certain embodiments, be configured to determine the property based on (at least) the absolute change in measurement signal and / or the rate of change of the signals. The control unit may incorporate the property determination unit and / or the signal processing unit or may be in addition to one or both ofthese. The property unit and / or signal processing unit may each have the form of a processor or controller as set out above for the control unit.Method and System

[0112] The system may be configured to perform any of the method steps disclosed herein. Moreover, any of the embodiments set out herein with respect to the method apply equally to the system, and any of the embodiments set out herein with respect to the system apply equally to the method.Pathways and individually controllable regions

[0113] A pathway refers to a defined route or channel through medium when provided at the sample surface of the system. It may be delimited by the manipulation assembly, for example by the extent of the force generated by the manipulation assembly within the medium (e.g. where provided by a field, by the field within the medium). This may be limited by the physical structure of the manipulation assembly, for example where there are electrodes for providing the force, the respective pathway may be delimited by the outermost electrodes of the manipulation assembly. Thus, the pathway may be determinable without medium being present, but is nevertheless formed within the medium when provided on the channel. The pathway is continuous, such that a force can be generated by the respective manipulation assembly across the length. The pathway may also be defined or delimited by structures within a medium, such as a separate medium region (e.g. of a different composition) or by channels or barriers within the medium. Accordingly, the pathway serves as conduit for the movement or separation of components of a sample within the systems and method. The pathways may be, but need not be, a linear pathway extending in a single direction. For example, the pathway may extend in plural directions. In some embodiments, the pathways may be physical channels. These may have a linear shape or may be non-linear, such as curved (e.g. serpentine).

[0114] The individually controllable region is a separately and independently actuatable region within and a part of the respective pathway. Each region extends only along a part of the respective pathway. The respective pathway may therefore comprise the individually controllable region. Sample may be moved through along the respective pathway, including the controllable region, using the respective manipulation assembly. The force, which is local to the individually controllable region, can be varied compared to the remainder of the respective pathway. This provides fine control over movement of the sample (or a portion thereof) along the respective pathway, leading to greater control over the manipulation and, where present, the separation process.

[0115] The second pathway is adjacent to or intersects the first pathway. Where a third pathway is present, it is adjacent to or intersects the second pathway. By “adjacent to” it is meant that the pathway may be next to and either spaced apart from or abut the other pathway. Accordingly, a component or a part thereof may move directly from one pathway to the other or there may be a spacing between these parts. The space may be through a portion of the medium such that the component(s) do not need to be removed from the medium. By “intersects”, it is meant that the pathway in question joins the otherpathway such that they are connected and may overlap. Thus, movement along the pathway in question may in turn result in removal from the earlier pathway.Sample

[0116] “Sample” refers to any material introduced into the system or used by the method for the purpose of separating it into its components. The sample can be in the form of a solid or a solution, for example, and is expected to comprise multiple components. This may be a matrix plus at least one other component (e.g. molecules or other substances of interest) or it may be plural components. For example, it may comprise a carrier (such as a liquid) and an analyte or component. It will be appreciated that a sample may not contain the anticipated or target components or analytes. As used herein, a sample can be separated into at least one component, where present. By “component”, it is meant one or a plurality of species within the sample. A component may be a single species, such as a single molecule, cell or other species, or it may be a mixture of these. Where a component is a mixture of plural species or analytes, a component may be further separated into sub -components, which also may be single species, such as a single molecule, cell or other species, or it may be a mixture of these. The component or, where present, sub-components may each be a target analyte of the system and methods. As referred to herein, a “portion” or “part” of a sample may be a portion of the whole sample (i.e. representative of the whole sample) or may be an isolated component or sub-component of the sample.

[0117] The analyte may, for example, be selected from a molecular species, a metal ion, a virus, and a microorganism. Biomolecule analytes are particularly useful and may, for instance, be a hormone selected from an eicosanoid, a steroid, an amino acid, amine, peptide, protein, a nucleic acid, single or double stranded DNA, peptide nucleic acid.Specific Uses

[0118] The systems and methods disclosed herein can be utilised in a number of different ways and fields. Some examples are provided below.

[0119] The systems and methods disclosed herein can be used to separate components from a mixture . The techniques disclosed herein advantageously allow for removal of unwanted components to allow for accurate analysis. In biological mixtures, there are often numerous components which can prevent or interfere with analysis. For example, high abundance components, such as molecules or proteins, to allow for better resolution for the remaining sample analysis.

[0120] In one embodiment, the systems and methods may be configured such that the separation step is an electrophoresis separation. That is, the system is configured to separate a sample containing charged species (e.g. particles, molecules, ions, etc.) based on their size and / or electrical charge. This can be used, for example, to separate DNA, RNA, carbohydrates or proteins. Accordingly, where this technique is used , and electric field can provide the force applied and movement of the sample through the medium causes separation. In further implementations, there may be a further separation, for example, using a different separation technique. For example, a first separation can take place usingelectrophoresis (e.g. along the first or second pathway) and a further separation of the components of the sample separated in the electrophoresis step can take place (e.g. along the second or a further pathway, where present) (“two dimensional” or “2D” separation, compared to a single (i.e. “one dimensional” or “ID” separation)). The second separation can be by isoelectric point, for example, where a pH gradient is formed in the matrix. This can be particularly useful for separation of proteins. The separation(s) can be further improved by the use of the individually controllable regions. For example, for the electrophoresis separation, complex force profiles can be created to provide greater separation between components and to create higher resolution. Using the modification element, further pH changes can be made along the pathway (s) creating opportunities for further manipulations based on isoelectric point. Traditional electrophoresis can suffer from overlap of spots and lack of resolution. Where there are two separations, such as using electrophoresis and isoelectric point, there may be different medium regions. A first in which there is a pH gradient, for example, (e.g. an IPG gel) and a second for isoelectric separation (e.g. an IEF gel). These may be continuous with the first pathway extending through the first region and the second extending through the first and into the second to permit transfer from the first pathway along the second pathway. There may be a plurality of second pathways extending perpendicular to the first corresponding to different components separated along the electrophoresis pathway. An alternative system and method may comprise a separation based on isoelectric point as a first separation and a subsequent electrophoresis separation (e.g. PAGE).

[0121] The systems and methods may be used for separating the products of a particular chemical reaction. For example, the systems and methods may be combined with techniques to subsequently separate out the products and, in some cases, concentrate the products. For example, this can be a nucleic acid amplification technique, for example polymerase chain reaction (PCR) amplification or an isothermal amplification technique (for example, loop mediated isothermal amplification (LAMP). The systems and methods of the present disclose are particularly suited for performing this type of reaction and separation because of the structure of the disclosed system and the ease at which thermal devices, such as heaters, can be integrated into the structures and because of the nature of the products produced (e.g. charged species). In some embodiments, the system may further comprise a reactor unit (this may be the modification element, or comprise the modification element) on which a reaction can occur, optionally wherein the reactor unit comprises a thermal device. The first manipulation assembly may be configured to move the sample from the reactor unit along the first pathway. The components can be separated (e.g. other components within the sample, unreacted reagents (e.g. primers)). The components can be further manipulated and, in some cases, concentrated at a particular point. They can also be analysed in situ using these. For example, the sensing elements set out herein advantageously can detect the resultant amplification product and, advantageously, can do so without necessarily requiring a tag, such as a fluor-labelled prove or intercalating dye. This can also be a quantitative measure, in that the electrical signal will be proportional to the amount of amplification product. Other examples where these systems and methods are advantageous include proteomic analysis, peptidomicanalysis, single cell studies, biomarker identification between disease and normal state, quality control for biological products, investigation of protein / DNA / RNA interactions, and analysis of carbohydrates (for example, separation and identification). For example, this can be used as part of an N glycosylation analysis where glycans are separated based on size and charge using the disclosed systems and methods.

[0122] In some embodiments, there may be multiple separate reactor units or sites. In the context of a nucleic acid amplification technique, different primers can be provided on each of the separate reactor units or sites to provide a multiplex assay. The resultant products can then be combined and manipulated using the disclosed systems or may each be transferred along separate (e.g. parallel) pathways.

[0123] The above systems could be combined with a third manipulation assembly and a third pathway. For example, so as to permit transfer of the separated components.

[0124] Another exemplary use is in cell manipulation and analysis. This can be for, example, used in cell lysis and / or flow cell analysis. A sample comprising cells (and, in some cases, other biological material) can be separated out with the cells individually analysed. This can be achieved based on e.g. the physical properties of the cell (density or size), on cell affinity (magnetic or binding properties) or on phenotypical properties (including e.g. the -omics at the cell surface). The cells can then be caused to migrate along a pathway and manipulated using a modification element. For example, for analysis, they can be held in a position along the pathway using a pair of electrodes (i.e. a modification element) on top of a sensing element (e.g. an electrode). Measurement of a property (e.g. impedance) can then take place using the sensing element. Alternatively or additionally, a manipulation can take place using the modification element or an additional modification element (which can be the same sensing element), for example the application of a voltage or current to cause lysis of the cell or electroporation to deliver material into the cell. Examples of the use of such systems and methods include in isolation of cells from blood (e.g. circulating tumour cells (“CTCs”)) and tests with LNCaP cells where both can be separated based on size from their respective samples. For example, these cells can be isolated and electrical lysis of their cell membranes can take place.

[0125] As noted above, the systems and methods can use electrowetting to manipulate a sample or a part thereof. One example of the use of such a technique would be in respect of PCR, where electrowetting would facilitate a sample droplet moving to mix with a PCR master mix (nucleotides, proteins, salts) and then subsequently moved to a region containing primers for the desired target.

[0126] The systems and method disclosed herein are advantageous in separation of complex mixtures, such as bodily fluids or created during cell lysis. Such mixtures often require several rounds of sample preparation, whereas the current systems and methods may be used to achieve separation with minimal operator interaction. For example, bodily fluids (such as blood, urine, saliva) contain large numbers of proteins which may be of limited use. The present systems can separate these out along a pathway or plural pathways, if required, leaving only the desired targets. The desired targets may be further separated as a clean sample, retained or held on a particular part of the device and / or moved separately for further analysis.Specific Implementations

[0127] Referring to FIGS. 1 and 2A to 2C, a system 100 for separating and manipulating samples comprises a separation apparatus 105 and a control unit 150 for operating the sensing chip. The separation apparatus 105 is used to receive and process samples containing a plurality components.

[0128] FIGS. 2A to 2C provide a schematic top view of the separation apparatus 105 chip in more detail. As show, the apparatus 105 comprises a substrate 108 defining a sample surface for receiving a medium and a sample within the medium, and through which medium the sample can be moved.

[0129] The apparatus 105 comprises a first manipulation assembly 113 comprising a set of electrodes comprised of five field electrodes 112A-E provided on the sample surface and arranged across the width of the apparatus 105. All of the field electrodes 112A-E have a separate contact control point and can be connected in various combinations to operate with another one of the field electrodes 112A-E, such that each can be actuated individually by the control unit 150 so that an electric field can be formed between any combination of the field electrodes 112A-E. This will cause any charged species within a sample provided on the sample surface to interact with the generated electric fields. The outermost field electrodes 112A, 112E on either side of the apparatus 105 accordingly define the first pathway 110 therebetween (see arrow A of Fig. 2B). An electric field formed between the outermost field electrodes 112A, 112E will cause charged species within the sample to move along the first pathway 110 (e.g. arrow A of Fig. 2B) between these outermost field electrodes 112A, 112E.

[0130] The intermediate field electrodes 112B-D located between outermost field electrodes 112A, 112E provide the first pathway 110 with multiple individually controllable regions. Specifically, each of these intermediate field electrodes 112B-D can either form an electric field with one of the outermost field electrodes 112A, 112E or with another of the intermediate field electrodes 112B-D thereby creating a local electric field which only extends across a part of the first pathway 110. These individually controllable regions accordingly allow for selective application of local electric field to influence or cause migration of sample portions within specific areas of the first pathway 110. For example, different combinations of the field electrodes 112A-E leads to fields at different points.

[0131] The separation apparatus 105 further includes a second manipulation assembly 123 defining four separate parallel second pathways 120A-D, with each of the second pathways 120A-D starting at a point along the first pathway 110 and extending in a direction perpendicular to the first pathway 110. In particular, the second manipulation assembly 123 comprises with four separate sets of electrodes arranged in parallel to one another each set comprising a first electrode 122A, 122C, 122E, 122G located at the top of the apparatus 105 and arranged side-by-side along the length of the first pathway 110 and a second electrode 122B, 122D, 122F, 122H located at the bottom of the apparatus 105 arranged side- by-side along the width of the apparatus 105 so as to oppose the corresponding first electrode 122A, 122C, 122 E, 122G of the respective pair. Each set of electrodes (i.e. first electrode 122A and second electrode 122B, first electrode 122C and second electrode 122D, etc.) is connected via a trace 124 and each pair is individually actuatable by the control unit 150 to generate an electric field therebetween atthe sample surface such that a charged species located in medium provided on the sample surface between each set is caused migrate along the respective second pathway 120A-D. As noted above, the sets of electrodes are arranged with the first pathway 110 extending between each pair of electrodes 122A-H so that a sample along the first pathway 110 can be moved from the first pathway 110 to and along the second pathway 120 by generation of an electric field therebetween. Operation of the second manipulation assembly 123 can be across all of the sets of electrodes so as to apply a (separate) force to cause at least a part of the sample to migrate along each of the second pathways 120A-D.

[0132] The arrangement of the second pathways 120A-D in relation to the first pathway 110 enables multi-dimensional separation and manipulation of components of a sample. For example, components separated along the first pathway 110 based on one property (such as size or charge) may be further separated or manipulated by directing them into different second pathways 120A-D. Each second pathway 120A-D may provide distinct separation conditions, allowing for enhanced resolution of complex mixtures.

[0133] The separation apparatus 105 also comprises include a plurality of sensing elements arranged in rows and columns for providing sensing capabilities the first pathway 110 and along each of the second pathways 120A-D.

[0134] In particular, the apparatus 105 comprises sixteen sensing elements located in the substrate 108 and exposed to the sample surface. The electrodes are arranged as four rows of four individual electrodes with a first row of electrodes 130A-D extending along the first pathway 110 with one electrode arranged between each neighbouring field electrode 112A-E of the first manipulation assembly 113. There are three further rows (a second row of electrodes 131A-D, a third row of electrodes 132A-D and a fourth row of electrodes 133 A-D) aligned with the first so as to define, together with the first row of electrodes 130A-D, four columns of electrodes with each column extending along a respective second pathway 120. In other words, there is a first column of electrodes 131A-133A (corresponding to one second pathway 120A); a second column of electrodes 131B-133B (corresponding to one second pathway 120B); a third column of electrodes 131C-133C (corresponding to one second pathway 120C); and a fourth column of electrodes 131D-133D (corresponding to one second pathway 120D). It will be appreciated that the overlap of the first pathway 110 with each of the second pathways 120A-D means that the first row of electrodes 130A-D are present in both the first pathway 110 and one of the second pathways 120A-D.

[0135] Although not depicted, each of the electrodes 130A-D, 131A-D, 132A-D, 133A-D can be configured with an appropriate set up for determining a property of a particular analyte or for measuring a particular property (or properties) of a sample. As such, each of the electrodes 130A-D, 131A-D, 132A-D, 133A-D may have a different configuration on the surface, such as a different specialized surface coatings or structures designed to selectively interact with specific sample components, such as an ion-selective membrane or capture species located thereon.

[0136] As set out above, the apparatus 105 also includes the intermediate field electrodes 112B-D located along the first pathway 110 to define individually controllable regions. These intermediate field electrodes 112B-D are positioned along the first pathway 110 and are configured to generate local forces within the localised areas between the field electrodes 112A-E when operated by the control unit 150. For example, the control unit 150 may selectively apply voltages to pairs of these intermediate field electrodes 112B-D together with one of the other field electrodes 112A-E to create localized electric fields that influence sample migration along the first pathway 110.

[0137] Accordingly, in use, a sample 101 may be positioned in the first pathway 110, as shown in FIG 2A. The first manipulation assembly 113 is used to apply an electric field so as to apply a force to cause charged species within sample 101 to migrate with the electric field. The control unit 150 may generate varying force profiles along the first pathway 110 using the local forces in the individually controllable regions. This may allow for precise control over sample migration. For instance, the control unit 150 may apply different voltage combinations to create varying electric field strengths or patterns along different portions of the first pathway 110.

[0138] As an example, the outermost field electrodes 112A, 112E of the first manipulation assembly 113 can be actuated by applying a voltage across the two outermost field electrodes 112A, 112E so as to generate a single electric field across the length of the first pathway 110 which electric field causes charged species in the sample to move in the direction of arrow A of Fig. 2B along the length of the first pathway 110. Furthermore, in this example, the medium and field are selected so that the charged species of the sample 101 will separate out along the length of the first pathway 110, for example on the basis of size and charge (for example, this can be using electrophoresis).

[0139] Further, the control unit 150 is configured to additionally provide local forces to the sample or components thereof by individually actuating pairs of the field electrodes 112A-E, either simultaneously or in addition to the overall force applied across the first pathway 110 as a separate force (e.g. after the force across the entire first pathway 110 has been removed). This capability allows for fine control over sample migration within specific regions of the first pathway 110, for example, by either reversing the field to move a particular component towards another part of the first pathway 110. This could be to align the sample with a particular sensing element (i.e. one of electrodes 130A-D) and / or with one of the second pathways 120A-D. This could also be used to amply the separation between certain components. For example, in the situation where two components are partially separated in the region on the second from right field electrode 112D but not sufficiently to avoid contamination, the second from right field electrode 112D could be actuated together with the middle field electrode 112C to drive one of the components back towards the start of the first pathway 110 (the opposite direction to Arrow A) and / or the second from right field electrode 112D could be actuated together with the outermost right field electrode 112E to cause the other component to move towards the end of the first pathway 110 (in the direction of Arrow A). Alternatively or additionally, the local forces can be used to slow down separation or speed up separation within a particular region. Forexample, either with the field between the outermost field electrodes 112A, 112E applied or not, a local force can be applied between the second from right field electrode 112D could be actuated together with the middle field electrode 112C. Depending on the polarity, this could either speed up or slow down the progression of sample (or components thereof) through the region between these two particular field electrodes 112D, 112E.

[0140] Alternatively or additionally, and dependent on the specific sample and charged species contained in the sample 101, the first manipulation assembly 113 can be actuated by applying a voltage across the various combinations of the field electrodes 112A-E so as to generate a plurality of local, different electric fields across the length of the first pathway 110. The local electric fields may each be different to have different effects on the sample 101 - and may be optimised based on ideal separations identified in calibration for a particular sample type - but the presence of continuous fields across the length of the pathway cause charged species in the sample 101 to move in the direction of arrow A of Fig. 2B along the length of the first pathway 110. In this example, because of the array of different local forces being applied across the length of the first pathway 110, the separation of the sample 101 will have a different profile to that of the example in which the outermost field electrodes 112A, 112E provide a single field applying a single force across the length of the first pathway 110. Accordingly, different voltage combinations applied to the field electrodes 112A-E can generate localized fields at various points along the first pathway 110. This enables more sophisticated manipulation of the sample. It will be appreciated that this may be combined with additional, separate local manipulation using individual pairs of the field electrodes 112A-E, as discussed above.

[0141] Regardless of the particular profile applied, the 101 and its components can be analysed using the row of electrodes 130A-D provided along the length of the first pathway 110. This can be for monitoring the separation and migration of the sample 101 along the length of the first pathway 110 and / or it can be to analyse the components after the separation is complete. For example, properties such as concentration, charge, size, or other characteristics. In addition, this information can be used by the control unit 150 to control operation of the apparatus 105. As an example, the control unit 150 may be configured to coordinate the application of voltages by the first manipulation assembly 113 and the second manipulation assembly 123 based on the readout from the electrodes 130A-D. This may allow for adaptive control of the separation process based on real-time measurements of sample properties.

[0142] For example, use of the row of electrodes 130A-D can be used during the operation of the apparatus 105 to provide signals indicative of properties of sample components along the pathways the respective first or second pathway 110. The signal from each electrode 130A-D can be provided to the control unit 150, which can then determine a property (such as the presence of a component at a particular point) and dynamically adjust the electric field applied to the sample (either across the first pathway 110 as a whole or within the individually controllable region, parameters or determine when specific components have reached desired locations along the pathway. In some implementations, the signals from electrodes 130A-D may be used to determine when to actuate the second manipulationassembly 123 to transfer separated components to the second pathways 120A-D. For instance, detection of a component at electrode 130C may trigger application of the second manipulation assembly 123 to provide a voltage across the field electrodes 122E, 122F provided either side of electrode 130C to move the identified component along the second pathway 120C. Or if the control unit 150 detects that a particular component of interest has reached a certain position along the first pathway 110, it may adjust the voltages applied by the first manipulation assembly to optimize the separation. Alternatively or additionally, the control unit 150 may use the determined property or properties to create feedback loops for continuous optimization of the separation and manipulation processes for future separations. This ability to monitor separation in real-time and selectively manipulate specific components accordingly allows for precise control over the separation process.

[0143] Alternatively or additionally, the control unit 150 may be configured to receive and process signals from the electrodes 130A-D to determine properties of the components located on the electrodes 130A-D prior to any further migration (e.g. along the second pathway 120). Based on this information, the control unit 150 may dynamically adjust the operation of the first manipulation assembly 113 and second manipulation assembly 123.

[0144] Regardless of the trigger for the actuation of the second manipulation assembly 123, use of the apparatus 105 can then comprise actuating the four separate pairs of electrodes to cause migration of components of the second pathways 120A-D. This can be simultaneously or separately or any combination, as required.

[0145] In particular, the control unit 150 causes each pair of electrodes 122A-H defining each of the second pathways 120A-D to generate a field therebetween so that electric fields are generated along each of the second pathways 120A-D. Any charged component(s) arranged along the respective part of the first pathway 110 can be moved from the region of the first pathway 110 overlapping the respective second pathway 120A-D along the second pathway 120 in the direction of the arrows shown in Fig. 2C. Depending on the desired outcome and the sample 101, the same or different electric fields can be applied to each of the second pathways 120. Further, depending on the specific use case, the electric field and the medium in these regions may be selected to cause a further separation of the components into sub-components (where present) along the second pathway 120.

[0146] In a similar manner to the sensing elements along the first pathway 110 (i.e. electrodes 130A- D), the electrodes 130A-D, 131A-D, 132A-D, 133A-D along each of the second pathways 120A-D can be used to monitor the progression of the manipulations / separations and / or to analyse the components or sub-components located along the respective second pathways 120A-D. The signals generated can be used in a similar manner to those generated along the first pathway 110.

[0147] Accordingly, it will be appreciated that this arrangement of field electrodes and can enable complex multidimensional separation and manipulation of samples. The first pathway 110 may provide movement and separation in one dimension, while the second pathways 120A-D may provide additional dimensions for further manipulation or analysis of separated components. The grid-like arrangement offield electrodes and sensing elements may provide flexibility in creating individually controllable regions along the first and second pathways 110, 120A-D. By selectively applying voltages to different combinations of electrodes, the control unit 150 may generate localized electric fields to influence sample migration within specific areas of the apparatus 105.

[0148] It will be further appreciated that modifications to this system and the method of using this system can be made.

[0149] For example, the configuration and location of the field electrodes 112A-E along the first pathway 110 is such that the control unit 150 can operate these to perform additional functions. For example, the field electrodes 112A-E may also serve as sensing elements in some implementations and / or have a manipulation function and thus serve as modification elements. For instance, the field electrodes 112A-E can be used to interrogate the sample 101 or a portion thereof located between any of the two field electrodes 112A-E. For example, it may be that the sample 101 will change the permittivity of the medium located therebetween and the field electrodes 112A-E are calibrated or arranged so that the change in permittivity can be determined using two of the field electrodes 112A-E. Similarly, it will be appreciated that although the electrodes defining the sensing elements have been discussed in the context of sensing elements that these could further be operated to act as a modification element, depending on the specific configuration of the control unit 150. For example, how these are used (e.g. voltage applied) and when these are used can determine how these interact with the various components. In some cases, the surfaces of these electrodes, etc. or the medium in which the sample is applied can be modified to allow these to act as modification elements as well as or instead of sensing elements.

[0150] Further, although not depicted, it may be that the medium provided on the sample surface of the substrate 108 is homogenous across the sample surface. Alternatively, there may be different medium regions. For example, the medium region in which the first pathway 110 is defined may have a first medium composition (and, for example, be configured for a first type of separation) and the medium region through which the (remainder of) the second pathways 120 extend may have a second medium composition (configured for a second type of separation). There may be further differences within each of these regions. For example, the region across the first pathway 110 may have a property gradient across the surface (for example, a pH gradient).

[0151] It will be appreciated based that the electrodes of system 100 could be used in other ways than those set out above to apply the forces. For example, these could be used to carry out application of a force according to any of the other types (set out as (i) to (v) above, or combinations thereof). For example, the electrodes 112A-112E could be used together with the medium to create concentration or pH gradients along the first pathway 110. This could then be used to separate charged species as a force is applied to a sample introduced to the medium.

[0152] Although not depicted, it will be appreciated that there may be additional electrodes along the second pathways 120. For example, there may be electrodes intersecting each of the electrodes 130A-133A and further along each of the second pathways 120. These may not be connected by the specific trace 124 arrangement depicted in Figs. 2A-C; instead, each may have a separate contact control point and can be connected in various combinations to operate with another one of the other intersecting electrodes, in a similar manner to electrodes 112A-112E.

[0153] Referring to FIGS. 3A-3C, various voltage profiles that may be applied by the control unit 150 and first manipulation assembly 113 in the system 100 have been exemplified. FIG. 3A depicts a graph showing the applied force on the first pathway 110 by plotting electric field strength (V / m) against position along the first pathway 110 (“D”). The positions of the field electrodes 112A-E of the first manipulation assembly 113 are labelled on the x-axis. The graph of FIG. 3 A depicts an arrangement in which the outermost field electrodes 112A, 112E of the first manipulation assembly are actuated by applying a voltage across the two outermost field electrodes 112A, 112E so as to generate a single electric field across the length of the first pathway 110. This is represented by the first field strength labelled as Vi. Further, in this example, an individual pair of field electrodes 112A-E is actuated. Specifically, the second from right field electrode 112D is actuated together with the middle field electrode 112C to provide an additional force applied in this particular region (depicted as V2), which has a greater field strength than the force across the entire first pathway 110. These can be applied either simultaneously or in addition to the overall force applied across the first pathway 110 by the first manipulation assembly 113.

[0154] FIG. 3B depicts a graph showing how voltage profiles V could be applied to field electrodes 112A-E over time (T) . In this example and in a similar manner to the example of Fig . 3 A, a first voltage V3 is applied across the two outermost field electrodes 112A, 112E so as to generate a single electric field across the length of the first pathway 110. This is applied from To to Ti . After the elapsed time T 1 , the voltage is stopped and a voltage in a particular individually controllable region (V4) is applied from time Ti to T2. The voltage V4 is negative and therefore represents an example in which species may be moved in a different direction during this second time period as compared to the first time period.

[0155] FIG. 3C depicts a graph showing a different applied force on the first pathway 110 to that of FIG. 3A. The graph of FIG. 3C also plots electric field strength (V / m) against position along the first pathway 110 (“D”). The positions of the field electrodes 112A-E of the first manipulation assembly 113 are labelled on the x-axis. In this graph, an arrangement is shown in which there is a stepped field strength increase provided across the length of the first pathway 110 by forming a plurality of local forces between each of the field electrodes 112A-E. Here a first electric field (strength V5) is generated by applying a voltage across the first two (from left to right) field electrodes 112A, 112B so as to generate an electric field across the first quarter of the first pathway 110. A second electric field (strength Ve which is greater than V5) is generated by applying a voltage across the second from left and middle field electrodes 112B, 112C so as to generate an electric field across the second quarter of the first pathway 110. Similarly, a third electric field (strength V7 which is greater than Ve) is generated by applying a voltage across the middle and second from right field electrodes 112C, 112D so as togenerate an electric field across the third quarter of the first pathway 110. Finally, a fourth electric field (strength V? which is greater than Ve) is generated by applying a voltage across the second from right and right most field electrodes 112D, 112E so as to generate an electric field across the fourth quarter of the first pathway 110. These can be applied in a stepwise manner, for example, as the sample migrates across the first pathway 110.

[0156] FIG. 4 provides a schematic depiction of a system 200 for separating and manipulating a sample 201. The system 200 comprises an apparatus 205 (e.g. a chip) and a control unit 250. As with the apparatus 105, the apparatus 205 is for separating and analysing samples containing multiple components. The control unit 250 is operatively connected to the apparatus 205 and is configured to operate the apparatus 205, including controlling sample movement and manipulation, the separation processes and data acquisition. The apparatus 205 comprises a substrate 208 on which the various components are arranged and which in part defines a sample surface 208A on which medium 202 is provided.

[0157] The apparatus 205 also comprises a first manipulation assembly 213 having a similar form to the first manipulation assembly 113 of the apparatus 105 of FIGS. 1 to 2C. Specifically, the first manipulation assembly 213 comprises a set of electrodes comprised of a plurality of field electrodes 212A-D located across the length of the substrate 208. A first field electrode 212A is located on one side of the substrate 208 defining one end of a first pathway 210 and the fourth field electrode 212D is located on the other side of the substrate 208 and defining the other end of the first pathway 210. Each of the field electrodes 212A-D have a separate contact control point and can be connected in various combinations to operate with another one of the field electrodes 212A-D. For example, the first and fourth field electrodes 212A, 212D can be used to generate an electric field across the first pathway 210 so that a sample 201 can be migrated across the length of the first pathway 210 and split into its components 201 A.

[0158] The intermediate (i.e. second and third) field electrodes 212B, 212C located between the first and fourth field electrodes 212A, 212D provide the first pathway 210 with multiple individually controllable regions. Each of the second and third field electrodes 212B, 212C can either form an electric field with one of the first or fourth field electrodes 212A, 212E or with the other intermediate (i.e. second or third) field electrode 212B, 212C thereby creating a smaller electric field which only extends across apart ofthe first pathway 210. These individually controllable regions accordingly allow for selective application of local electric field to influence or cause migration of sample portions within specific areas ofthe first pathway 210.

[0159] The apparatus 205 also includes four sensing elements 230A-D arranged in a row along the length ofthe first pathway 210. The sensing elements 230A-D are each comprised of a pair of opposing electrodes, with each electrode of the pair provided on either side of first pathway 210. Each sensing element 230A-D is individually addressable so that it can provide its own measurement signal. In this way, properties of the sample 201 or its components 201 A can be detected as it moves through themedium 202 along the first pathway 210. The measurement signals generated by the sensing elements 230A-D are provided to the control unit 250 which can determine a property of the sample 201 or its components 201 A. As set out above, this data can be used to monitor the progression of the separations / manipulations or to analyse components 201 A of the sample 201.

[0160] One example arrangement comprises the sensing elements 230A-D extending out of the sample surface 208A so that the medium 202 is located therebetween. Sample 201 and / or a component 201 A moving along the first pathway 210 will accordingly pass directly between the pair of electrodes (some or all of these, depending on progression along the first pathway 210). The measurement signal can accordingly be an indication of impedance, conductivity or resistance as the sample 201 or a component 201 A moves between the pair of electrodes of each sensing element 230A-D.

[0161] The apparatus 205 is further provided with a first recess 235A and a second recess 235B provided in the substrate 208 along the first pathway 210 and each extend from an opening in the sample surface 208A to abase. The first recess 235A is arranged along the first pathway 210 at a position where a particular component 201 A of interest is expected to separate out of the sample 201. The second recess 235B is arranged further along the first pathway 210 at a position where another particular component 201 A of interest is expected to separate out of the sample 201. Depending on the specific implementation, medium 202 may extend into the recesses 235A, 235B (this can be the same type of medium 202 as that provided on the sample surface 208A or there may be a different type, forming a second medium region) or there may be no medium in the recesses 235A, 235B. The use of the first and second recesses 235A, 235B is set out in more detail, below.

[0162] The apparatus 205 also includes a second manipulation assembly 223 defining two separate, parallel second pathways 220A, 220B (visible in FIG. 5B). In a similar manner to the second pathways 120A-D of the apparatus 105 of FIGS. 2A and 2B, the second pathways 220A, 220B of this apparatus 205 extend perpendicularly to the first pathway 210 and extend from various points along the length of the first pathway 210 where the first and second pathways 210, 220A, 220B intersect; however, one significant difference here is that the second pathways 220A, 220B extend in the z-axis (relative to the plane defined by the sample surface) - i.e. perpendicular to the sample surface. In particular, the second manipulation assembly 223 comprises a set of electrodes comprising four field electrodes 222A-D forming two pairs of electrodes. The first pair comprises a first field electrode 222A located above the medium 202 and directly above the first recess 235A and a second field electrode 222B electrically connected to the first field electrode 222A via a wire 214 and located below the substrate 208 directly below the first recess 235A. This defines one second pathway 220A and generation of an electric field between the first and second field electrodes 222A, 222B of the second manipulation assembly 223 causes sample 201 or a component 201 A thereof to migrate in the z-axis between the first and second field electrodes 222A, 222B. Given the location of the first and second field electrodes 222A, 222B and the first recess 235A, this can be used to drive a sample 201 or a component 201A thereof (as shown by the arrow in FIG. 5B) along this particular second pathway 220A from part of the medium 202located above the first recess 235A into the first recess 235A. The second pair of electrodes of the second manipulation assembly 223 comprises a third field electrode 222C located above the medium 202 and directly above the second recess 235B and a fourth field electrode 222D electrically connected to the third field electrode 222D via a wire 214 and located below the substrate 208 directly below the second recess 235B. This defines the other second pathway 220A and generation of an electric field between the third and fourth field electrodes 222C, 222D of the second manipulation assembly 223 causes sample 201 or a component 201 A thereof to migrate in the z-axis along between the third and fourth field electrodes 222C, 222D. This can be used to migrate a sample 201 or a component 201 A thereof (as shown by the arrow in FIG. 5B) along this particular second pathway 220B from part of the medium 202 located above the second recess 235B into the second recess 235B. Accordingly, the arrangement of the second pathways 120A-D in relation to the first pathway 210 enables multidimensional separation and manipulation of components of a sample 201. Components 201 A separated along the first pathway 210 may be further separated or manipulated by directing them into the different second pathways 220A, 220B.

[0163] The use of the first and second recesses 235A, 235B provide for improved handling and analysis. For example, the first and second recesses 235A, 235B provide a region in which the components 201 A can be collected. This can provide for improved direct analysis, since there is less likely to be interference when performing measurements in the first and second recesses 235A, 235B from other components 201 A, the medium 202 or the environment around the apparatus 205. Components 201 A collected in the first and second recesses 235A, 235B may be extracted from the system for further processing or characterization, such as sequencing. The components 201 A can, for example, be concentrated by collection in the base of the first and second recesses 235A, 235B. In such cases, the substrate 208 may be configured as a substrate 208 which can be used for further analysis directly. For example, components 201 A collected in the first and second recesses 235A, 235B may be analysed on the substrate 208 using techniques such as mass spectrometry, Raman spectroscopy, or optical methods. The first and second recesses 235A, 235B may also serve as interfaces for coupling to external instruments or microfluidic components.

[0164] The apparatus 205 also comprises further sensing elements in the form of a first electrode 236A located in the substrate 208 and which surrounds the base of the first recess 235A and a second electrode 236B also located in the substrate 208 and which surrounds the base of the second recess 235B. Corresponding counter electrodes (not shown) and reference electrodes (not shown) may be present in the substrate 208. The first and second electrodes 236A, 236B can interrogate components 201 A received in the corresponding first and second recesses 235A, 235B. It will be appreciated that other sensing elements or systems could be used.

[0165] Although in the above embodiment, the pairs of electrodes forming the sensing elements 230A-D are used for sensing, it will be appreciated that these could be further used by the control unit 250 for other purposes, for example, as modification elements.

[0166] A further system 300 is depicted in the top view of FIG. 6. The system 300 depicted therein comprises a control unit (not shown) and an apparatus 205 which is operated by the control unit. The system 300 has similar functionality to the systems 100, 200 of FIGS. 1 to 5B, but with additional functionality for performing reactions (such as DNA amplification) and with additional modes of separation.

[0167] The apparatus 305 comprises a substrate 308 on which a medium is provided. The substrate 308 in part defines a sample surface along which the samples analysed using system 300 can be migrated.

[0168] The apparatus comprises four first manipulation assemblies 313A-B arranged as four parallel rows from the top (as depicted in Fig. 6) to the bottom of the substrate 308. Each first manipulation assembly 313A-D has a similar structure to the first manipulation assemblies 113, 213 of the systems 200, 300 of FIGS. 1 to 5B in that it each first manipulation assembly 313A-D comprises a set of electrically connected electrodes comprised of a plurality of field electrodes 312A, 312B extending across the width of the substrate 308. For each first manipulation assembly 313 A-D a first field electrode 312A, 312C, 312E, 312G provided on the outermost left hand side of the substrate 308 as depicted in Fig. 6 delimits a first pathway 310 and a second field electrode 312B, 312D, 312F, 312H provided on the outermost right hand side of the substrate 308 delimits the opposite end of the corresponding first pathway 310. An electric field formed between the first and second field electrodes 312A-H of each first manipulation assembly 313A-D will cause charged species within the sample to move along the corresponding first pathway 320 between the corresponding first and second field electrodes 312A-H (i.e. from right to left or left to right, as depicted in FIG. 6).

[0169] Between the first and second field electrode 312A-H of each first manipulation assembly 313A-D are further electrodes defining individually controllable regions and the second pathways 320. In particular, the apparatus 305 comprises a grid of electrode arrays 322A-D which in this embodiment is a 4x4 grid comprises of four rows of electrode arrays 322A-D and four columns of electrode arrays 322A-D. Along each first pathway 310 is provided one row of four electrode arrays 322A-D (only those of the first row are labelled in FIG. 6, for the sake of clarity), with the four electrode arrays 322A-D of each of the first pathways 310 aligned so as to further form four columns of four electrode arrays 322A- D moving down the substrate 308 across the first pathways 310, as depicted in FIG. 6.

[0170] Each electrode array 322A-D itself comprises four electrodes arranged so that one of the four electrodes defines the edge of a square. Fig. 7 shows an expanded view of a part of the apparatus in which the first electrode array 322A arranged at the top left of the apparatus 305 as it is shown in Fig. 6 (i.e. the first electrode array 322A in the 4 x 4 grid) is more clearly visible. Here it can be seen that the first electrode array 322A comprises four elongated electrodes 322A1-A4 arranged in a square shape. This provides two sets of opposing pairs of electrodes. Specifically, there is a first pair comprising a first electrode 322A1 arranged to extend across the uppermost first pathway 310. The first electrode 322A1 opposes and is spaced apart from a third electrode 322A3 (i.e. defining the oppositeside of the square), which is also arranged to extend across the uppermost first pathway 310. The first electrode 322A1 and third electrode 322A3 of the first electrode array 322A are electrically connected to one another so that an electric field can be formed therebetween. There is also a second pair comprising a second electrode 322A2 arranged to extend along the top edge of the uppermost first pathway 310 (in this arrangement, between the top ends of the first electrode 322A1 and third electrode 322A3). This opposes and is spaced apart from a fourth electrode 322A4 (i.e. defining the opposite side of the square), which is also arranged to extend across the bottom edge of the uppermost first pathway 310. The second electrode 322A2 and fourth electrode 322A4 of the first electrode array 322A are electrically connected to one another so that an electric field can be formed therebetween. Each of the electrode arrays 322A-D of the apparatus 305 has a corresponding structure with a pair formed of a first and third electrode 322A1, 322A3 extending across one of the first pathways 310 and a pair formed a second and fourth electrode 322A2, 322A4 extending along the edge of one of the first pathways 310.

[0171] It will be appreciated that the electrode arrays 322A-D ofthe apparatus 305 can serve multiple purposes and, accordingly, the control unit may be configured to operate these for a number of different reasons and operations during the use of the apparatus 305 and system 300.

[0172] The arrangement of the electrode arrays 322A-D along each first pathways 310 provides four separate pairs of first and third electrodes 322A1, 322A3 which extend across each first pathway 310. Each of these pairs of first and third electrodes 322A1, 322A3 can be actuated to provide a local electric field across a portion of the respective first pathway 310 to thereby influence or cause migration of a sample along the respective first pathway 310 within an individually controllable region.

[0173] Furthermore, in some embodiments, each of the electrode arrays 322A-D may be further electrically connected to at least one of the first and second field electrodes 312A-F delimiting the corresponding first pathway 310 along which the electrode array 322A-D in question is located. This may be the first electrode 322A1, the third electrode 322A3 or both the first and third electrode 322A1, 322A3. This creates additional individually controllable regions between the various combinations of electrodes located along and defining each first pathway 310.

[0174] Moreover, each of the first and third electrodes 322A1, 322A3 of each electrode array 322A- D can be further electrically connected to the first and third electrodes 322A1, 322A3 along the row forming a part of the same first pathway 310 and, further, the each of the first and third electrodes 322A1, 322A3 of each electrode array 322A-D align within the respective column defined by the electrode arrays 322A-D.

[0175] The apparatus 305 also comprises four second manipulation assemblies 323A-D defining four parallel second pathways 320. In particular, the alignment of four electrode arrays 322A-D in a line perpendicular to the first pathways 310 (i.e. in the columns across the surface of the substrate 308 as per the depiction of FIG. 6) and with the electrode arrays 322A-D either abutting or overlapping with their immediate neighbours creates four second manipulation assemblies 323A-D and, accordingly, four separate second pathways 320 which extend from the second electrode 332A2 of each electrode array322A-D along the top of the substrate 308 to the fourth electrode of each of the electrode arrays along the bottom of the substrate 308. Each of these pairs of second and fourth electrodes 322A2, 322A4 can be actuated to provide a local electric field across a portion of the substrate 308 and together each column of neighbouring electrode arrays 322A-D defines a respective second pathway 320 to thereby influence or cause migration of a sample across the length along the respective second pathway 320. For example, the four leftmost electrode arrays starting from the top left electrode array 322A as depicted in FIG. 6 and finishing with the bottom leftmost array (which forms part of the bottommost first manipulation assembly 313D) form one second pathway 323 A and a sample may be drawn continuously along the entire length of this second pathway 323A using the electric fields generated between each of the second and fourth electrodes of these electrode arrays. Moreover, as a result of this structure each individual electrode array along each the second pathways 310 also provides its own an individually controllable region.

[0176] The apparatus 305 in this embodiment further comprises a plurality of sensing elements 335A- 335D (only the top four are labelled in FIG. 6 for the sake of clarity) in the form of electrodes provided in the substrate 308, in a similar manner to the system 100 of FIGS. 2A to 2C but having a 4 x 4 grid structure corresponding to the electrode arrays 322A-D. In particular, each of the first pathways 310 is provided with four sensing elements 335A-335D along the length, with one sensing element 335A- 335D associated with and located at the centre of each electrode array 322A-332D along the length of the corresponding first pathway 310. Due to the alignment of the electrode arrays 322A-D forming each second pathway 320, it follows that each second pathway 320 also comprises four sensing elements 335A-335D provided along its length. Each sensing element 322A-D provides a means for determining a property provided along each of the first and second pathways 310, 320. As set out above, this can be used for the purpose of analysis of the components of a sample and / or for real-time changes to the separation or manipulation processes.

[0177] The apparatus 305 may further comprise reactors 307A-D for use in a reaction or, more generally, a conversion of one entity to another. For example, the reactors 307A-D may be heaters on which a reaction mixture can be received and which can be used to instigate a reaction. One example is the reagents for PCR.

[0178] The apparatus 305 includes four reactors 307A-D with one reactor 307A-D arranged at one end of each of the first pathways 310. In this way, the result of a reaction performed using the reactors 307A-D can be used as the sample(s) for the subsequent separation and manipulations performed by the system 300 and the corresponding first manipulation assembly 312 can be used to draw each sample along the first pathway 310 away from the respective reactor 307A-D. Depending on the specific implementation, medium may extend over the reactors 307A-D or there may be an additional medium region.

[0179] The arrangement of these reactors 307A-D along parallel and interconnected (via the second pathways 320) first pathways 310 allows for various manipulations. For example, similar separationscan occur along each of the first pathways 310 from the reaction products from reactors 307A-D. Depending on the conditions and the samples, these could be separated so that like components are aligned along the first pathway 310 and in line with a respective second pathway 320. For example, a component A in each sample could be aligned with the first column defined by the lefthand second pathway 320. The second manipulation assembly 323A can be actuated (using the electrode arrays 322 A) to cause each component A in each first pathway 310 to accumulate at one particular point, thereby collecting and concentrating the component A. This can be useful where analysis is improved with greater recovery of component A. This actuation can be informed by the sensing elements 335 A- D and / or the sensing elements may be used to provide information on the properties of the sample.

[0180] Alternatively, and although not depicted, collection sites, such as recesses or regions at the end of one of the first or second pathways 310, 320, could be provided for collection of a particular component after separation.

[0181] It will be further be appreciated that the grid structure of the electrode arrays 322A-D and the field electrodes 312A-H is such that a sample could be manipulated and separated in more than two dimensions - i.e. along more than two pathways. For example, it is possible for a sample to be manipulated or separated along a first pathway 310, followed by movement or separation along a second pathway 320. At this point, the control unit may be further configured to cause one of the other first manipulation assemblies to cause the component to be moved along another first pathway 310 (providing a third pathway). Accordingly, the system 300 provides a multi-dimensional manipulation system with a vast degree of flexibility to perform complex separations and manipulations.

[0182] As an alternative to, or in addition to the above, the apparatus 305 may be modified so as to replace or supplement the sensing elements 335A-D with recesses. In particular, one modification of the apparatus 305 would be one in which it comprises a plurality of recesses in place of or in addition to the sensing elements 335A-335D. The recesses can be formed in the substrate 308, in a similar manner to the system 200 of FIG. 5A and 5B but having a 4 x 4 grid structure corresponding to the electrode arrays 322A-D. In particular, each of the first pathways 310 in this modification is provided with four recesses along the length, with one recess associated with and located at the centre of each electrode array 322A-332D along the length of the corresponding first pathway 310 (i.e. the recess would each correspond to where each sensing element 335 A-D is depicted in FIGS. 6 and 7). Due to the alignment of the electrode arrays 322A-D forming each second pathway 320, it follows that each second pathway 320 also comprises four recesses provided along its length. Each recess extends from an opening in the sample surface to a base (not depicted), in a similar manner to that of FIGS. 5A and 5B. Depending on the specific implementation, medium may extend into the recesses (this can be the same type of medium as that provided on the sample surface or there may be a different type, forming a second medium region) or there may be no medium in the recesses.

[0183] In this modified embodiment, the apparatus also includes a third manipulation assembly which defines a plurality of third pathways extending in the z-axis (relative to the plane defined by the samplesurface) - i.e. perpendicular to the sample surface. The third manipulation assembly comprises a first plate electrode located above the medium and directly above the recesses and a second plate electrode electrically connected to the first plate electrode and located below the substrate directly below the recesses. Generation of an electric field between the first and second plate electrodes of the third manipulation assembly causes sample or a component thereof to migrate in the z-axis between the first and second plate electrodes. Given the relative positioning of the first and second plate electrodes and the recesses, this can be used to drive a sample or a component thereof along a third second pathway from the medium on the top of the sample surface into a recess. The first and second plate electrodes, the medium and each recess therefore defines its own third pathway. Although not depicted, the apparatus 305 may further comprise sensing elements in or around each recess for in

[0184] Returning to the example in which a component A in each sample generated by the reactors 307A-D could be aligned with the first column defined by the lefthand second pathway 320, the second manipulation assembly 323A can be actuated (using the electrode arrays 322A) to cause each component A in each first pathway 310 to accumulate at one particular point, thereby collecting and concentrating the component A. Once in this position, component A could be passed through the openings in the sample surface into a respective recess.

[0185] This modified embodiment may be combined with the sensing elements 335A-D discussed above. For example, the sensing elements 335A-D may surround the openings in the sample surface. This can also be useful for informing actuation of the third manipulation assembly. Alternatively or additionally, a sensing element may be provided along each recess. As an example, an electrode or electrode pair may surround the base of each recess. These can be used to interrogate components received in the recesses.

[0186] One specific implementation using a system such as the system 300 depicted in FIGS. 6 and 7 is for an amplification process, such as PCR. Either a reaction mixture including PCR primers and a sample could be provided to each site, or a different reaction mixture including different primers (but in some cases the same sample) could be provided to each reactor site. The amplification can be driven by the reactors 307A-D. Given the structure of the apparatus 305, it lends itself to traditional semiconductor manufacturing techniques and enables the formation of the apparatus 305 as an integrated circuit. This can give close control over the reactor conditions, enabling fast and accurate heating cycles for PCR. Different cycles could be performed at different reactors 307A-D, if required. Once the amplification is complete, each first manipulation assembly 313 A-D is configured to move the sample from the respective reactor 307A-D along the respective first pathway 310. The components can be separated (e.g. other components within the sample, unreacted reagents (e.g. primers)) and the target component(s) (i.e. the amplification product(s)) can be isolated. In embodiments with sensing elements 335 A-D in the pathways, the target component(s) can be detected using the sensing elements 335A-D. For example, a sensing element 335A-D may be able to detect a respond corresponding first to unreacted primers, then to partially reacted molecules and then the target component(s). This caninform how a reaction has proceeded and how the separation is occurring. The target component(s) can also be isolated along the respective second pathways 320 and / or concentrated by combining the same target component(s) in a single region on the substrate 308.

[0187] FIGS 8A and 8B provide exemplary systems 400, 400’ which can utilise plural apparatus 405 A-C (for example, which can have the structure of any of the apparatus 105, 205, 305 disclosed herein) for manipulating and separating samples. FIG. 8A provides a system 400 comprising a fluidly interconnected arrangement of apparatuses 405. In this particular structure, there is a first apparatus 405A which itself is fluidly connected to at least one second apparatuses 405B (in this embodiment, two second apparatuses 405B) via at least one fluid conduit 406. In turn, each of the second apparatuses 405B is connected to at least one third apparatus 405C (in this embodiment, two separate third apparatuses 405 C, each) via at least one fluid conduit 406. The fluid conduits 406 may serve as an input or output conduit for fluid samples. For example, the fluid conduits 406 may be provided with the output of a separation or manipulation process and provide this to the second or third apparatuses 405B, 405C for further separation and manipulation.

[0188] A control unit 450 is operably connected to each to control the various components within each. The control unit 450 accordingly may coordinate and manage the operation of the first, second and third apparatuses 405A-C. This may include controlling fluid flow, processing parameters, and data acquisition. For example, the control unit 450 may be programmed to adjust reaction conditions locally at specific first, second and third apparatuses 405A-C based on real-time data received from sensing elements integrated into the first, second and third apparatuses 405A-C.

[0189] In such a systems 400, 400’, each apparatus 405A, 405B, 405C may be responsible for specific tasks such as sample separation, reaction staging, or monitoring of biochemical reactions. The arrangement allows for parallel processing or sequential operations on fluid samples, enhancing throughput and efficiency. Moreover, the hierarchical or tree-like structure of FIG. 8A can facilitate complex fluid sample processing tasks, including complex synthesis pathways. This may therefore employ apparatuses such as that depicted in FIG. 6 provided with reactors.

[0190] Exemplary uses include sample processing. A sample may be separated into separate components and then the components moved to a separate second or third apparatuses 405B, 405C for a reaction. Alternatively or additionally, a sample may be separated a first voltage at the first apparatus 405A and then at a second voltage at one of the second or third apparatuses 405B, 405C. Different components can be provided to different second and third apparatuses 405B, 405C.

[0191] A further system 400’ having a similar structure is shown. In this system, there are four apparatuses first apparatuses 405A which are fluidly connected to one another via at least one fluid conduit 406. The first apparatuses 405A in this system 400’ are connected in a chain, providing a further configuration for specific types of sample separation and preparation.

[0192] Fig. 9 depicts a further system 500 for separating and manipulating samples. The system 500 comprises a substrate, which in this embodiment can be formed of a plurality of stacked layers 551,554, 555, 557 arranged as an integrated chip. The structure and function of these stacked layers 551, 554, 555, 557 can be selected based on the application in mind, as set out in more detail below.

[0193] On the sample surface defined by the upper surface of the substrate is provided a medium comprised of three separate portions or regions: a first medium region 502A extending across the depth of the substrate (from front to back as depicted in FIG. 9) and approximately one third across the width of the substrate; a second medium region 502B is also provided on the sample surface and covers the remainder of the upper sample surface with a height equal to that of the first medium region 502A; and a third medium region 502C is provided on top of the second medium region 502B and comprises the same dimensions as the second medium region 502B so as to cover the second medium region 502B. These provide three different environments in which migration and separation can occur.

[0194] A first manipulation assembly 513 is provided at the first medium region 502A in the form of a set of electrodes (not visible) provided in the substrate at either end and along the length of the first medium region 502A (i .e . from the front surface of the first medium region 502A to the back as depicted in Fig. 9). The electrodes of the first manipulation assembly 513 accordingly define a first pathway 510 extending through the first medium region 502 A and can cause migration of a sample (and separation, where required) along the length of the first medium region 502A. The electrodes positioned along the length of the first medium region 502 A provide individually controllable regions along the length of the first pathway 510.

[0195] A second manipulation assembly 523 is provided at the first and second medium regions 502A, 502B and defines a second pathway 520 extending perpendicular to the first pathway 510 across the width of the substrate as shown in Fig. 9. The second manipulation assembly 523 comprises a first field electrode (not visible) provided in the substrate at the first medium region 502A on the outer side of the first pathway 510 and the second field electrode (not visible) is provided in the substrate at the opposite side of the second medium region 502B. In this way, components 501 A separated out from the sample along the first pathway 510 can be transferred from the first pathway 510 and the first medium region 502A to the second medium region 502B along the second pathway 520, and subsequently through the second medium region 502B where a second separation of the components 501 A into sub -components 50 IB can take place.

[0196] A third manipulation assembly 543 is provided to form a third pathway 540 extending perpendicular to both the first and second pathways 510, 520 (the z-axis, where the first pathway 510 extends in the x-axis and the second pathway 520 extends in the y-axis). The third manipulation assembly 543 comprises a first field electrode 542B formed as a layer beneath the substrate and a second field electrode provided above the third medium region 502C on the opposite side of the device to the first field electrode 542B. The first field electrode 542A and second field electrode 542B are both formed as planar electrodes and the first field electrode 542A can be formed as an integral part of the integral circuit forming the substrate, if required. Sub-components formed along the second pathway 520 can be caused to migrate by upwards from the second medium region 502B into the third mediumregion 502C by forming an electric field between the first field electrode 542A and the second field electrode 542B.

[0197] A control unit (not depicted) can control operation of each of the first, second and third manipulation assemblies 513, 523, 543.

[0198] Such a system can be used for a number of different processes. For example, one such use could comprise a three-dimensional sample preparation method. In this example, the first medium region 502A comprises an IEF gel such that separation along the first pathway 510 is by isoelectric point. The second medium region 502B uses PAGE to separate the components 501A into subcomponents 50 IB using their electrophoretic mobility. The third medium region 502C can be a transfer membrane into which the sub-components 50 IB are transferred so that they can be removed from the system 500 with no direct handling from a user, for example to be analysed by another device.

[0199] In certain implementations of the system 500, the substrate may have a particular layer structure which may provide additional functionality.

[0200] In some specific implementation, the lowermost layer of the substrate (above the first field electrode 542A) can be an ASIC layer 554 which can communication (or in some embodiments, comprise) the control unit or a part thereof. Functional control and / or measure components 558 can be provided within the ASIC layer 554. Above the ASIC layer 554, is a via layer 551 providing interconnections between the ASIC layer 554 and the other layers of the substrate. Above the via layer 551 is a functional layer 555, which can be a microelectromechanical system (MEMS)Zintegrated passive device (IPD) layer. The functional layer 555 can comprise functional components 553 such as a heater and / or thermometer 553. The uppermost layer, which is above the functional layer 555 and which defines the sample surface, can be a flow cell layer 557. This provides a surface which can be removed and replaced so that the rest of the integrated circuit can be re-used. Components can be formed in the flow cell layer 557, for example a virtual thermal well 552 could extend from the sample surface to the thermometers 553, where present. These integrated electronic components may allow for thermal control and sensing within the system 500. For example, the virtual thermal well 552 and heater / thermometer 553 may be used to create and maintain specific temperature profiles along the pathways, which may influence sample migration or reaction kinetics. These can accordingly act as modification elements. The MEMS / IPD layer 557 may, in some cases, provide additional functionality such as micro-scale actuation or integrated photodetection for sample analysis. The ASIC layer 554 may provide control unit functionality, for example, by processing signals from the various sensing elements and relaying signals to control operation of the first, second and third manipulation assemblies 513, 523, 543 and other components.

[0201] FIGS. 10A-10B depict various specific parts of apparatuses which can be used in the systems and methods disclosed herein. FIG. 10A depicts a part of an apparatus in which cells 601, which have been separated out (e.g. using a first separation method, such as movement along a first pathway) are caused to migrate along a pathway 610 (under the influence of a force). The apparatus also comprisesa modification element 632A comprising a pair of electrodes which are operable to apply an electric field which retains the cells 601 in a particular position as they pass. The pair of electrodes are shown as providing confinement in the z-direction (i .e . perpendicular to the pathway 610 out of the substrate) . Associated with each modification element 632A is a sensing element 632B in the form of a planar electrode on the surface the cells 601 pass over. Cells 601 are held by the modification element 632A on sensing element 632B and can be interrogated. Compared to existing cell interrogation systems, such as those relying on hydrodynamic focusing, this provides continuous measurement, which can provide more reliable information about the cells, provide additional information using electrochemical measurement, is faster and puts less stress on cells. Of course, these components could be operated in a different manner under the control of the control unit, with the pair of electrodes operating as a sensing element rather than modification element 632A. Similarly, a manipulation can take place using the modification element 632A or the sensing element 632B, for example the application of a voltage or current to cause lysis of the cell

[0202] FIG. 10B depicts an arrangement for trapping ions 601 A from a sample in a particular position which can utilise electrodes 632C, 632D having a structure (e.g. as endcaps) such as that ofthe electrode arrays depicted FIGS. 6 and 7. The electrodes 632C can each generate a field which retains a positively charged particle surrounded by a cloud of similarly charged particles such that a particle 601 A received between four electrodes 632C, 632D can be trapped (i.e. retained). These accordingly can act as a modification element. The electric fields E which cause the trapping may be generated by a quadrupole of electrodes 632C (positive) and a ring electrode 632D. Fig 10B left shows a first state during an AC cycle and Fig. 10B right shows a second state during an AC cycle.

[0203] FIG. 11 provides a flowchart depicting a method 180 of separating and manipulating a sample comprising a plurality of components, the method comprising: providing 182 the sample comprising a plurality of components to a medium; applying a force 184 to the sample so as to cause the sample to migrate along a first pathway through the medium; and applying a force 186 to at least a part of the sample so as to cause the part of the sample to migrate from the first pathway along a second pathway through the medium, wherein the second pathway is adjacent to or intersects the first pathway; and wherein an individually controllable region is provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a first local force to the individually controllable region so as to influence or cause migration of at least a part of sample migrate along the respective first and / or second pathway within the individually controllable region. Variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the disclosed systems and methods, from a study of the drawings, the disclosure, and the appended claims. In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite article "a" or "an" does not exclude a plurality. A single processor or other unit may fulfil the functions of several items recited in the claims. The merefact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. If a computer program is discussed above, it may be stored / distributed on a suitable medium, such as an optical storage medium or a solid-state medium supplied together with or as part of other hardware, but may also be distributed in other forms, such as via the Internet or other wired or wireless telecommunication systems. If the term "adapted to" is used in the claims or description, it is noted the term "adapted to" is intended to be equivalent to the term "configured to". Any reference signs in the claims should not be construed as limiting the scope.

[0204] The flow diagrams and block diagrams in the Figures illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various embodiments of the disclosed systems and methods. In this regard, each block in the flow diagrams orblock diagrams may represent a module, segment, or portion of instructions, which comprises one or more executable instructions for implementing the specified logical function(s). In some alternative implementations, the functions noted in the block may occur out of the order noted in the figures. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality involved. It will also be noted that each block of the block diagrams and / or flow diagrams and combinations of blocks in the block diagrams and / or flow diagrams, can be implemented by special purpose hardware-based systems that perform the specified functions or acts or carry out combinations of special purpose hardware and computer instructions.

[0205] Further examples of the methods and systems disclosed herein are provided as clauses

[0206] Clause 1. A system for separating and manipulating a sample comprising a plurality of components, the system comprising: a substrate defining a sample surface for receiving a medium in which the sample can be received; a first manipulation assembly operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway; and a second manipulation assembly operable to apply a force to a sample to cause the at least a part of the sample to migrate within medium provided at the sample surface along a second pathway, wherein the second manipulation assembly is arranged so that the second pathway is adjacent to or intersects the first pathway, wherein at least one of the first manipulation assembly and the second manipulation assembly is configured so as to define at least one individually controllable region along a part of the respective first and / or second pathway in which a first local force can be selectively applied within the individually controllable region so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region; and wherein the system further comprises a control unit configured to operate: the first manipulation assembly so as to cause the sample to migrate within medium provided at the sample surface along the first pathway; and the second manipulation assembly so as to cause at least a part of the sample to migrate within medium provided at the sample surface along the second pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into aplurality of components; and wherein the control unit is further configured to operate at least one of the first manipulation assembly and the second manipulation assembly to generate the first local force within the respective first and / or second pathway so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region.

[0207] Clause 2. The system of clause 1, wherein at least one of the first manipulation assembly and / or the second manipulation assembly is configured to provide a plurality of individually controllable regions along a part of the respective first and / or second pathway in which a respective local force can be selectively applied to at least a part of sample within the respective individually controllable regions.

[0208] Clause 3. The system of clause 2, wherein the control unit is configured to operate at least one of the first manipulation assembly and / or the second manipulation assembly so as to provide different force profiles along the respective first pathway and / or second pathway using the respective local forces in the plurality of individually controllable regions.

[0209] Clause 4. The system of any preceding clause, wherein the system is configured such that the first manipulation assembly is operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway so as to separate the sample into a plurality of components.

[0210] Clause 5. The system of any preceding clause, wherein the first manipulation assembly and second manipulation assembly each comprises a first electrode set arranged to define the respective first pathway and second pathway and to provide the respective force.

[0211] Clause 6. The system of clause 5, wherein at least one of the first manipulation assembly and / or second manipulation assembly further comprises at least one further electrode provided along the respective first pathway and second pathway which in part defines the individual controllable region; and wherein the control unit is configured to operate the at least one further electrode to generate the first local force.

[0212] Clause 7. The system of clause 6, wherein the at least one further electrode is further operable to provide a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway.

[0213] Clause 8. The system of clause 7, wherein the control unit is further configured to determine a property of the component based on the signal provided by the at least one further electrode, optionally wherein the control unit is further configured to control the first manipulation assembly and / or the second manipulation assembly based on the determination of a property of the component of the sample.

[0214] Clause 9. The system of any preceding clause, further comprising at least one modification element provided along the first and / or second pathway operable to: interact with at least a part of the sample migrating along the respective first and / or second pathway and / or influence the migration of the at least a part of the sample along the respective first and / or second pathway; and wherein the control unit is configured to operate the modification element so as to cause the modification element to interactwith and / or influence the migration of at least a part of the sample within the respective first and / or second pathway.

[0215] Clause 10. The system of the clause 9, wherein the modification element is provided on the substrate and located along the first pathway and / or second pathway.

[0216] Clause 11. The system of clause 10, wherein the modification element comprises an electrode operable to interact with at least a part of the sample.

[0217] Clause 12. The system of clause 11, wherein the electrode is provided with a surface comprising a capture agent configured to bind to at least one component of the sample such that the component can be retained on the electrode; and / or wherein the electrode is provided with a capture species on a surface thereof and configured to selectively bind to at least one component of the sample such that the component can be selectively retained on the electrode.

[0218] Clause 13. The system of any of clauses 9 to 12, wherein the control unit is configured to operate the modification element so as to retain at least one component of the sample thereon.

[0219] Clause 14. The system of any of clauses 9 to 13, wherein the system comprises a property modification unit comprising the modification element, the property modification unit is configured to modify a property of medium provided at the sample surface using the modification element so as to influence the migration of at least a part of the sample within the respective first and / or second pathway.

[0220] Clause 15. The system of clause 14, wherein the property modification unit is configured to modify at least one of pH, electrical conductivity, a thermal property or ionic strength of the medium.

[0221] Clause 16. The system of clause 14 or clause 15, wherein the property modification unit is configured to form a first property region across a first portion of one of the first pathway and / or second pathway and form a second property region across a second portion of the respective pathway such that the property varies across the respective pathway.

[0222] Clause 17. The system of any of clauses 9 to 16, wherein the property modification unit comprises a first electrode set operable to modify a property of the medium in the first property region and a second electrode set operable to modify a property of the medium in the second property region, wherein each of the first electrode set and the second electrode set comprises at least one modification element in the form of an electrode.

[0223] Clause 18. The system of any of clauses 9 to 17, wherein the modification element is a thermal device.

[0224] Clause 19. The system of any preceding clause, further comprising a sensing element operable to provide a signal indicative of a property of a component of the sample at a position along at least one of the first pathway and the second pathway.

[0225] Clause 20. The system of clause 19, wherein the control unit is further configured to determine a property of the component based on the signal provided by the sensing element.

[0226] Clause 21. The system of clause 20, wherein the control unit is further configured to control the first manipulation assembly and / or the second manipulation assembly based on the determination of a property of the component of the sample.

[0227] Clause 22. The system of any preceding clause, wherein the second manipulation assembly is further operable to cause the at least a part of the sample to migrate from the first pathway along the second pathway.

[0228] Clause 23. The system of any preceding clause, wherein at least one of the first manipulation assembly and second manipulation assembly is operable to: generate an electric field to provide the respective force; generate a magnetic field to provide the respective force; cause a property of the medium to vary along a pathway so as to provide a force which acts on at least a part of the sample so as to cause the sample or a part thereof to migrate through the medium; generate a centrifugal or centripetal force to provide the respective force; or generate acoustic waves to provide the respective force..

[0229] Clause 24. The system of any preceding clause, further comprising a third manipulation assembly operable to cause at least a part of the sample to migrate within medium provided at the sample surface along a third pathway, wherein the third manipulation assembly is arranged so that the third pathway is adjacent to or intersects the second pathway, wherein the control unit is further configured to control the third manipulation assembly so as to cause at least a part of the sample within medium provided at the sample surface along the third pathway.

[0230] Clause 25. The system of clause 24, wherein the third manipulation assembly is operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along the third pathway so as to separate the sample into a plurality of components.

[0231] Clause 26. The system of clause 24 or clause 25, wherein the third manipulation assembly is further operable to cause the part of the sample to migrate from the second pathway along the third pathway.

[0232] Clause 27. The system of any of clauses 24 to 26, wherein at least one of the first, second and third pathways extends at an angle relative to the sample surface, optionally wherein the first pathway extends in a first direction through the medium in a first plane, the second pathway extends in a second direction through medium in the first plane and at an angle relative to the first direction; and at least a part of the third pathway extends in a second plane intersecting and at an angle to the first plane.

[0233] Clause 28. The system of clause 27, wherein the substrate comprises an opening provided therein at the sample surface; and wherein at least one of the second pathway and the third pathway is arranged at an angle relative to the sample surface at the opening and extends into the opening so that migration of the at least a part of the sample causes migration through the opening.

[0234] Clause 29. The system of any preceding clause, further comprising a medium provided at the sample surface.

[0235] Clause 30. The system of clause 29, wherein the medium comprises: a first medium portion having a first medium property and through which at least a portion of the first pathway extends; and a second medium portion having a second medium property and through which at least a portion of the second pathway extends, wherein the first and second medium properties are different.

[0236] Clause 31. The system of any preceding clause, wherein migration along the first pathway is arranged to separate the sample into a plurality of components and wherein the second manipulation assembly is operable to cause a plurality of components to migrate from the first pathway, each component migrating along a separate second pathway adjacent to or intersecting the first pathway.

[0237] Clause 32. A method of separating and manipulating a sample comprising a plurality of components, the method comprising: providing a medium comprising a sample, the sample comprising a plurality of components; applying a force to the sample so as to cause the sample to migrate along a first pathway through the medium; and applying a force to at least a part of the sample so as to cause the part of the sample to migrate from the first pathway along a second pathway through the medium, wherein the second pathway is adjacent to or intersects the first pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components; and wherein an individually controllable region is provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a first local force to the individually controllable region so as to influence or cause migration of at least a part of sample migrate along the respective first and / or second pathway within the individually controllable region.

[0238] Clause 33. The method of clause 32, wherein a plurality of individually controllable regions are provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a local force to each of the individually controllable region so as to cause a part of sample to migrate along the respective first and / or second pathway within each of the plurality of individually controllable regions.

[0239] Clause 34. The method of clause 33, wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises generating different force profiles along the respective first pathway and / or second pathway using the plurality of individually controllable regions.

[0240] Clause 35. The method of any of clauses 32 to 34, wherein applying a force comprises at least one of applying an electric field to the sample or a part thereof; applying a magnetic field to the sample to a part thereof; applying a force may comprise providing a medium in which a property of the mediumvaries along a pathway so as to provide a force which acts on at least a part of the sample so as to cause the sample or a part thereof to migrate through the medium; applying a force to cause sedimentation of a sample or a part thereof; applying a centrifugal or centripetal force to the sample or a part thereof; or applying a force using acoustic waves.

[0241] Clause 36. The method of any of clauses 32 to 35, further comprising providing a first manipulation assembly operable to provide a force to the sample to cause the sample to migrate along the first pathway; and a second manipulation assembly operable to apply a force to the sample to cause the at least a part of the sample to migrate along the second pathway, wherein the first manipulation assembly and second manipulation assembly each comprises a first electrode set arranged to define the respective first pathway and second pathway and to provide the respective force.

[0242] Clause 37. The method of clause 36, wherein at least one of the first manipulation assembly and / or second manipulation assembly further comprises at least one further electrode provided along the respective first pathway and second pathway which in part defines the individual controllable region; and wherein the method comprises operating the at least one further electrode to generate the first local force.

[0243] Clause 38. The method of clause 37, further comprising obtaining a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway using the at least one further electrode.

[0244] Clause 39. The method of clause 38, further comprising determining a property of the component based on the signal provided by the at least one electrode, optionally further comprising controlling the step of applying the force to the sample or component along the first pathway and / or the second pathway based on the determination of a property.

[0245] Clause 40. The method of any of clauses 32 to 39, further comprising providing at least one modification element along the first and / or second pathway, wherein the at least one modification element is operable to interact with at least a part of the sample migrating along the respective first and / or second pathway and / or influence the migration of the at least a part of the sample along the respective first and / or second pathway; and wherein the method further comprises causing the modification element to interact with and / or influence the migration of at least a part of the sample within the respective first and / or second pathway.

[0246] Clause 41. The method of clause 40, wherein the modification element comprises an electrode operable to interact with at least a part of the sample.

[0247] Clause 42. The method of clause 41, wherein the electrode is provided with a capture species on a surface thereof and configured to selectively bind to at least one component of the sample such that the component can be selectively retained on the electrode.

[0248] Clause 43. The method of any of clauses 40 to 42, further comprising operating the modification element so as to retain at least one component of the sample thereon.

[0249] Clause 44. The method of any of clauses 40 to 43, wherein the method further comprises modifying a property of the medium so as to impart the variation in the medium property along at least a portion of the first pathway and / or the second pathway.

[0250] Clause 45. The method of clause 44, wherein modifying a property comprises forming a first property region across a first portion of one of the first pathway and / or the second pathway, and forming a second property region across a second portion of the respective pathway such that the property applied varies across the respective pathway.

[0251] Clause 46. The method of clause 45, wherein the method further comprises providing a property modification unit comprising a first electrode set operable to modify a property of the medium in the first property region and a second electrode set operable to modify a property of the medium in the second property region, and wherein modifying a property comprises actuating the first set of electrodes and second set of electrodes.

[0252] Clause 47. The method of clause 46, wherein the modification element is a thermal device and wherein the method comprises operating the thermal device to heat or cool the medium and / or a part of the sample.

[0253] Clause 48. The method of any of clauses 40 to 47, wherein the method comprises operating the modification element to apply a stimulus to the sample or a component of the sample so as to cause a transformation of the sample or the component.

[0254] Clause 49. The method of any of clauses 32 to 48, wherein causing the sample to migrate along the first pathway causes separation of the sample into a plurality of components and causing a part of the sample to migrate along the second pathway comprises causing at least one component to migrate along the second pathway, optionally further wherein causing a part of the sample to migrate along the second pathway causes separation of the component into a plurality of sub -components.

[0255] Clause 50. The method any of clauses 32 to 49, wherein applying a force so as to cause the sample to migrate along a first pathway comprises applying a plurality of forces across the first pathway such that the force applied to the sample varies across the first pathway, the plurality of force profiles comprising a first local force across the individually controllable region of the first pathway and a second force across a second portion of the first pathway.

[0256] Clause 51. The method of clause 50, further comprising providing a first electrode set comprising at least two electrodes arranged to provide the first local force at the individually controllable region and a second electrode set comprising at least two electrodes operable to provide the second force at the second portion.

[0257] Clause 52. The method of any of clauses 32 to 51, further comprising obtaining a signal indicative of a property of a component of the sample at a position along the first pathway, optionally wherein the position along the first pathway is a point at which the first pathway and the second pathway intersect.

[0258] Clause 53. The method of clause 52, further comprising determining a property of the component based on the signal provided, optionally further comprising controlling the step of applying the force to the sample or component along the first pathway and / or the second pathway based on the determination of a property.

[0259] Clause 54. The method of any of clauses 32 to 53, wherein applying a force so as to cause the sample to migrate along a second pathway comprises applying a first force profile across a first portion of the second pathway and applying a second force profile across a second portion of the second pathway such that the force applied varies across the second pathway.

[0260] Clause 55. The method of clause 54, further comprising providing a second manipulation assembly operable to provide the plurality of force profiles across the second pathway, wherein the second manipulation assembly comprises a plurality of electrodes operable to provide the plurality of force profiles across the second pathway and wherein at least one electrode of the plurality of electrodes is located along the second pathway.

[0261] Clause 56. The method of clause 55, wherein the second manipulation assembly comprises a first electrode set comprising at least two electrodes of the plurality of electrodes arranged to provide a force at the first portion and a second electrode set comprising at least two electrodes of the plurality of electrodes operable to provide a force at the second portion.

[0262] Clause 57. The method of any of clauses 32 to 56, further comprising obtaining a signal indicative of a property of a component of the sample at a position along the second pathway.

[0263] Clause 58. The method of clause 57, further comprising determining a property of the component based on the signal, optionally further comprising controlling the second manipulation assembly based on the determination of a property of the component of the sample.

[0264] Clause 59. The method of any of clauses 32 to 58, further comprising applying a force to at least a part of the sample so as to cause at least a part of the sample to migrate along a third pathway through the medium, wherein the third pathway is adjacent to or intersects the second pathway, optionally wherein applying a force to at least a part of the sample is so as to cause at least a part of the sample to migrate from the second pathway along the third pathway.

[0265] Clause 60. The method of clause 59, wherein migration along at least one of the first pathway or the second pathway is arranged to separate the sample into a plurality of components; and wherein migration along a third pathway comprises migrating at least one component from the second pathway along the third pathway, optionally wherein migration of the at least one component along the third pathway causes separation of the component into a plurality of sub -components.

[0266] Clause 61. The method of clause 59 or clause 60, wherein migration of at least a part of the sample along the third pathway comprises migrating the at least a part along the third pathway to an analysis location and determining at property of the at least a part at the analysis location.

[0267] Clause 62. The method of clause 60 or clause 61, wherein the medium is provided at the sample surface and wherein at least one of the first, second and third pathways extends at an anglerelative to the sample surface, optionally wherein the first pathway extends in a first direction through the medium in a first plane, the second pathway extends in a second direction through medium in the first plane and at an angle relative to the first direction; and at least a part of the third pathway extends in a second plane intersecting and at an angle to the first plane.

[0268] Clause 63. The method of any of clauses 60 to 62, further comprising providing a substrate defining the sample surface, the substrate comprising an opening provided therein at the sample surface; and wherein at least one of the second pathway and the third pathway is arranged at an angle relative to the sample surface at the opening and extends into the opening so that migration of at least a part of the sample causes movement of the at least a part of the sample through the opening.

[0269] Clause 64. The method of any of clauses 32 to 63, wherein the medium comprises: a first medium portion having a first medium property and through which at least a portion of the first pathway extends; and a second medium portion having a second medium property and through which at least a portion of the second pathway extends, wherein the first and second medium properties are different.

[0270] Clause 65. The method of any of clauses 32 to 64, wherein a causing a part of the sample to migrate along a second pathway through the medium comprises: causing a plurality of components to migrate from the first pathway to the second pathway, each component of the plurality migrating along a separate second pathway adjacent to or intersecting the first pathway.

[0271] Clause 66. The method of clause 65, wherein each second pathway is adjacent to or intersects a different point along the first pathway.

[0272] Clause 67. The method of any of clauses 32 to 66, further comprising providing a plurality of third pathways; and causing a part of the sample provided to each second pathway to migrate out of the respective second pathway along a corresponding third pathway within the medium, wherein each third pathway is adjacent to or intersects the respective second pathway.

Claims

CLAIMS:

1. A system for separating and manipulating a sample comprising a plurality of components, the system comprising: a substrate defining a sample surface for receiving a medium in which the sample can be received; a first manipulation assembly operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway; and a second manipulation assembly operable to apply a force to a sample to cause the at least a part of the sample to migrate within medium provided at the sample surface along a second pathway, wherein the second manipulation assembly is arranged so that the second pathway is adjacent to or intersects the first pathway, wherein at least one of the first manipulation assembly and the second manipulation assembly is configured so as to define at least one individually controllable region along a part of the respective first and / or second pathway in which a first local force can be selectively applied within the individually controllable region so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region; and wherein the system further comprises a control unit configured to operate: the first manipulation assembly so as to cause the sample to migrate within medium provided at the sample surface along the first pathway; and the second manipulation assembly so as to cause at least a part of the sample to migrate within medium provided at the sample surface along the second pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components; and wherein the control unit is further configured to operate at least one of the first manipulation assembly and the second manipulation assembly to generate the first local force within the respective first and / or second pathway so as to influence or cause migration of at least a part of sample along the respective first and / or second pathway within the individually controllable region.

2. The system of claim 1, wherein at least one of the first manipulation assembly and / or the second manipulation assembly is configured to provide a plurality of individually controllable regions along a part of the respective first and / or second pathway in which a respective local force can be selectively applied to at least a part of sample within the respective individually controllable regions; and wherein the control unit is configured to operate at least one of the first manipulation assembly and / or the second manipulation assembly so as to provide different force profiles along the respective first pathway and / or second pathway using the respective local forces in the plurality of individually controllable regions.

3. The system of claim 1 or claim 2, wherein the system is configured such that the first manipulation assembly is operable to apply a force to a sample to cause the sample to migrate within medium provided at the sample surface along a first pathway so as to separate the sample into a plurality of components.

4. The system of any preceding claim, wherein the first manipulation assembly and second manipulation assembly each comprises a first electrode set arranged to define the respective first pathway and second pathway and to provide the respective force.

5. The system of claim 4, wherein at least one of the first manipulation assembly and / or second manipulation assembly further comprises at least one further electrode provided along the respective first pathway and second pathway which in part defines the individual controllable region; and wherein the control unit is configured to operate the at least one further electrode to generate the first local force.

6. The system of claim 5, wherein the at least one further electrode is further operable to provide a signal indicative of a property of a component of the sample at a position along the respective first pathway and / or second pathway; and wherein the control unit is further configured to determine a property of the component based on the signal provided by the at least one further electrode.

7. The system of any preceding claim, further comprising at least one modification element provided along the first and / or second pathway operable to: interact with at least a part of the sample migrating along the respective first and / or second pathway and / or influence the migration of the at least a part of the sample along the respective first and / or second pathway; and wherein the control unit is configured to operate the modification element so as to cause the modification element to interact with and / or influence the migration of at least a part of the sample within the respective first and / or second pathway.

8. The system of claim 7, wherein the system comprises a property modification unit comprising the modification element, the property modification unit is configured to modify a property of medium provided at the sample surface using the modification element so as to influence the migration of at least a part of the sample within the respective first and / or second pathway.

9. The system of any preceding claim, further comprising a sensing element operable to provide a signal indicative of a property of a component of the sample at a position along at least one of the first pathway and the second pathway; and wherein the control unit is further configured to determine a property of the component based on the signal provided by the sensing element.

10. The system of any preceding claim, wherein the second manipulation assembly is further operable to cause the at least a part of the sample to migrate from the first pathway along the second pathway.

11. The system of any preceding claim, wherein the second manipulation assembly is operable to cause a plurality of components to migrate from the first pathway, each component of the plurality migrating along a separate second pathway adjacent to or intersecting the first pathway.

12. A method of separating and manipulating a sample comprising a plurality of components, the method comprising: providing a medium comprising a sample, the sample comprising a plurality of components; applying a force to the sample so as to cause the sample to migrate along a first pathway through the medium; and applying a force to at least a part of the sample so as to cause the part of the sample to migrate from the first pathway along a second pathway through the medium, wherein the second pathway is adjacent to or intersects the first pathway, wherein migration along at least one of the first pathway and the second pathway is arranged to separate the sample into a plurality of components; and wherein an individually controllable region is provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a first local force to the individually controllable region so as to influence or cause migration of at least a part of sample migrate along the respective first and / or second pathway within the individually controllable region.

13. The method of claim 12, wherein a plurality of individually controllable regions are provided along at least one of the first pathway and / or the second pathway and wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises selectively applying a local force to each of the individually controllable region so as to cause a part of sample to migrate along the respective first and / or second pathway within each of the plurality of individually controllable regions.

14. The method of claim 13, wherein applying a force to the sample so as to cause the sample to migrate along the respective first pathway and / or second pathway through the medium comprises generating different force profiles along the respective first pathway and / or second pathway using the plurality of individually controllable regions.

15. The method of any of claims 12 to 14, further comprising obtaining a signal indicative of a property of a component of the sample at a position along at least one of the first pathway and the second pathway and determining a property of the component based on the signal obtained.

16. The method of claim 15, wherein controlling the step of applying the force to the sample or component along the first pathway and / or the second pathway is based on the determination of a property.

17. The method any of claims 12 to 16, wherein applying a force so as to cause the sample to migrate along a first pathway comprises applying a plurality of forces across the first pathway such that the force applied to the sample varies across the first pathway, the plurality of forces comprising a first local force across the individually controllable region of the first pathway and a second force across a second portion of the first pathway.

18. The method any of claims 12 to 17, further comprising providing at least one modification element along the first and / or second pathway, wherein the at least one modification element is operable to interact with at least a part of the sample migrating along the respective first and / or second pathway and / or influence the migration of the at least a part of the sample along the respective first and / or second pathway; and wherein the method further comprises causing the modification element to interact with and / or influence the migration of at least a part of the sample within the respective first and / or second pathway.

19. The method any of claims 12 to 18, further comprising operating the modification element so as to retain at least one component of the sample thereon; and / or operating the modification element to apply a stimulus to the sample or a component of the sample so as to cause a transformation of the sample or the component.

20. The method of any of claims 12 to 19, wherein causing a component of the plurality of components to migrate from the first pathway along a second pathway through the medium comprises: causing a plurality of components to migrate from the first pathway, each component of the plurality migrating along a separate second pathway adjacent to or intersecting the first pathway.

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