NANOPORE ARRAY WITH ELECTRODE CONNECTORS PROTECTED FROM ELECTROSTATIC DISCHARGE
20210300750 · 2021-09-30
Inventors
Cpc classification
B81B7/0022
PERFORMING OPERATIONS; TRANSPORTING
B01L3/5085
PERFORMING OPERATIONS; TRANSPORTING
G01N33/48721
PHYSICS
B01L2300/161
PERFORMING OPERATIONS; TRANSPORTING
International classification
B81B7/00
PERFORMING OPERATIONS; TRANSPORTING
B01L3/00
PERFORMING OPERATIONS; TRANSPORTING
Abstract
A component (8) adapted to engage with a receiver (6) has an array of contact pads (16) to removeably connect with a corresponding array of connectors (18) on the receiver (6). Each contact pad (16) of the array is electrically connected to the electrode (26) of a corresponding recess or well (28) that is part of a sensor, wherein a membrane is formable across each recess. A conductive grid (102) is configured between the contact pads (16) of the array, to inhibit an electrostatic discharge (ESD) conducting across the recesses or wells and/or direct an ESD away from the recesses or wells.
Claims
1. A component configured to removably engage with a receiver, the component having: an array of electrodes and configured to removably connect with a corresponding array of connectors on a receiver, wherein a plurality of electrodes of the array are electrically connected to a corresponding recess or well, each recess forming part of a sensor for receiving a fluid therein such that a membrane is formable across each recess that separates the fluid in a recess from a conductive fluid in a common chamber; a conductive structure configured across the array to enable an electrical connection between the array of electrodes and an array of connectors of a receiver, wherein the conductive structure is configured to inhibit an electrostatic discharge (ESD) conducting across the recess and/or direct an ESD away from the recess.
2. A component according to claim 1, further having a conductive fluid in the common chamber and fluid in the plurality of recesses, wherein a membrane is formed across a plurality of the recesses separating the fluid in the common chamber from the fluid in each of the plurality of recesses, and wherein the conductive structure is configured to inhibit an electrostatic discharge passing across a membrane via the recess and/or direct an electrostatic discharge away from a membrane.
3. A component according to claim 2, wherein the conductive structure is connected to the fluid in the common chamber to inhibit a potential difference occurring across the recess or membrane.
4. A component according to claim 2 further comprising a common electrode in contact with the fluid in the common chamber.
5. A component according to claim 4 wherein the conductive structure is electrically connected to the common electrode.
6. A component according to claim 2 wherein the membrane is an amphipathic bilayer.
7. A component according to any preceding claim, wherein the array of electrodes is arranged on a base substrate and the conductive structure is mounted on the substrate.
8. A component according to any preceding claim, wherein the conductive structure extends from a plane defined by the electrodes.
9. A component according to claim 7, wherein the array of electrodes is arranged on a base of substrate and wherein at least part of the substrate extends from the region between the electrodes to form a wall and the conductive structure is configured on top of the wall.
10. A component according to claim 7 or 8, wherein the conductive structure is formed from the deposition of a conductive material in the region between electrodes.
11. A component according to any preceding claim, wherein the conductive structure is configured as a grid and extends in a planar direction, defined by the base or substrate, between the electrodes.
12. A component according to any preceding claim, wherein the electrodes are arranged in an array having a rectilinear pattern.
13. A component according to any preceding claim, wherein the footprint of each electrode is quadrilateral.
14. A component according to any preceding claim, wherein the pitch of the electrodes of the array is between 100 um and 1500 um.
15. A component according to any preceding claim, wherein the thickness of the walls of the conductive structure, in cross-section, is between 20 um and 200 um.
16. A component according to any preceding claim, wherein the windows or apertures of the conductive structure through which the connectors extend have rounded corners.
17. A component according to any preceding claim, wherein the electrode array and conductive structure are covered, at least in part, with a removable protective film or protective layer.
18. A kit having a component according to any preceding claim and a receiver, wherein the array of connectors on the receiver are configured to extend through the conductive structure without contacting the conductive structure to form an electrical connection with the electrode on the component.
19. A kit according to claim 18, wherein the grid is connected to the conductive fluid in the sample chamber to inhibit a potential difference occurring across the recess or membrane formed between the fluid in the sample chamber and fluid in a recess, such that when a conductive fluid occupies the sample chamber an ESD is inhibited from conducting through, or in the region of, the recess and/or sensor by the conductive structure.
Description
[0041] The invention is discussed below, by way of example only, with reference to the following figures in which:
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[0060] In
[0061] The component 8 can be disposable and, by way of example, has a disposable flow-cell located therein. The flow cell can be equivalent to that discussed in WO 2014/064443, which is hereby incorporated in its entirety by reference, wherein the component is configured to be a removable low-cost component, which can be disposed of after a single use. This is achieved by configuring more expensive components of the device 4 within the receiver 6. The low-cost component makes it feasible to perform multiple experiments with different flow-cells relatively cheaply. The base component 10 can house the electronics and cooling configuration for the overall apparatus 2. The receiver 6 can house further electronics not included in the base 10 and functions as an adaptor to receive the component 8.
[0062] Electrical connections are known from WO2009/077734, which is hereby incorporated in its entirety by reference and provides an example of the usage of the ‘solder bump’ approach that provides an electrical connection to a layer of amphiphilic molecules.
[0063] The well 28 is formed in the substrate 24 of non-conductive material and can be used to form or support a layer of amphiphilic molecules. In use, an aqueous solution can be introduced to the well 28, and region therearound, such that a layer of amphiphilic molecules is formed across the well 28 separating the aqueous solution in the well 28 from the remaining volume of aqueous solution above the recess. The arrangement of the well or recess 28, the well electrode 26 and additional circuitry (not shown) enables measurement of electrical signals across a layer of amphiphilic molecules. The well electrode can make electrical contact with the aqueous solution in the recess 28.
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[0065] The recess or well 28 forms part of a sensor, and the sensing components must communicate with a reader or microprocessor. In the present invention these wells reside in the component 8 and must electrically communicate with a reader on the receiver 6 and/or base 10.
[0066] Removable connections, or non-permanent electrical connections are known, by way of example, from WO2016/059417, which is hereby incorporated in its entirety by reference and is suitable for an array of electrical connections in such a way that the component parts can be attached and detached, and optionally reattached thereafter, without requiring extreme conditions (whether chemical or environmental) to trigger the connection or disconnection.
[0067] While removably detachable electrical connections are known the inventor has realised that conventional connections for parts such as the component 8 and receiver 6 can be improved. In particular the interface can be improved to provide an additional layer of protection to the well or recess 28 and any sensor or sensing component residing or formed in such a recess 28. The interface that is connected to the wells on the component is sensitive to mechanical and/or electrical shock.
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[0071] The invention is particularly suited to the protection of membranes formed over wells or recesses that are susceptible to damage or rupture by uncontrolled voltages, such as ESD, flowing through the well or sensor. The membrane may be a layer formed from amphiphilic molecules such as phospholipids, which have both hydrophilic and lipophilic properties. The amphiphilic layer may be a monolayer or a bilayer. The amphiphilic layer may be a co-block polymer such as disclosed in Gonzalez-Perez et al., Langmuir, 2009, 25, 10447-10450 or WO2014/064444, hereby incorporated by reference in its entirety.
[0072] The membrane may comprise an aperture formed in a solid state layer, which may be referred to as a solid state pore. The aperture may be a well, gap, channel, trench or slit provided in the solid state layer along or into which analyte may pass. Such a solid-state layer is not of biological origin. Solid state layers can be formed from both organic and inorganic materials including, but not limited to, insulating materials such as Si3N4, Al203, and SiO, organic and inorganic polymers such as polyamide, plastics such as Teflon® or elastomers such as two-component addition-cure silicone rubber, and glasses. The solid state layer may be formed from graphene. Suitable graphene layers are disclosed in WO-2009/035647, WO-2011/046706 or WO-2012/138357. Suitable methods to prepare an array of solid state pores is disclosed in WO-2016/187519.
[0073] A biological nanopore may be provided in one or more of the membranes providing a conduction pathway across each membrane which serves to fluidically connect fluid provided in a well of the well array with solution provided in an upper chamber. The nanopore may be a transmembrane protein pore derived from but not limited to alpha-hemolysin, anthrax toxin and leucocidins, outer membrane proteins/porins of bacteria, such as Mycobacterium smegmatis porin & lysenin. The pore may be derived from CsgG, such as disclosed in WO-2016/034591. The nanopore may be provided in the aperture of a solid state membrane. Such pores are known as hybrid pores. The nanopore may be formed from DNA origami.
[0074] The device of the invention is particularly suited for estimating the sequence of a polymer analyte. The analyte may be for example a polynucleotide, a polypeptide or a polysaccharide. Measurement of the polymer may take place during translocation of the polymer through the nanopore under a potential difference applied across the nanopore. The measurement may be a measurement of ion flow through the nanopore during translocation.
[0075] Ionic solution may be provided in the wells in contact with each respective membrane and the electrode provided in each of the respective wells of the array.
[0076] The component 8 can be supplied with a polar medium forming a meniscus or membrane over the well 28, having a nanopore located in the membrane of each well 28. The meniscus cooperates with the well to form part of the sensor. Alternatively, the component can be supplied ‘dry’ and without a polar medium, which is added to the component before installation to the device 6 and prior to testing or analysis of a sample. Further still, the component can be configured with a solid-state membrane and/or solid-state.
[0077] The meniscus and/or sensors of the well are sensitive to uncontrolled voltages, such as ESD. The wells, therefore, must be protected by inhibiting a voltage discharge passing across or through the well. In use, the pads 100 of the array of electrodes 16 are exposed in preparation for engagement with the array of connectors 18 on the device. Although the component 8 can be connected to a device 6 without the pads being touched, they remain susceptible to accidental touch by, for example, a finger tip. In the field, which is the antithesis of controlled laboratory conditions, a user would typically remove a component from its packaging and manually place it in the receiver. The invention mitigates any detrimental effects of a user contacting the array of electrodes 16 with, for example, their finger tip.
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[0079] Additionally or alternatively the grid can be connected to the polar medium 32, or the chamber in which the polar medium resides—this connection can be by means of at least one of: a dedicated via; a wired or bonded connection to a portion of a substrate; an electrical connection through the body of the component 8; or an electrical connection to the structure forming the chamber holding the polar medium 32, which surrounds the wells 28 of the array 16.
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[0081] The grid can also be connected to the ground plane of the component, and the device, once the component is installed in the device 6. In other words, if the substrate is conductive and coated in a dielectric or insulator, the grid can dissipate charge into this capacitance, whose field affects both common and well electrodes.
[0082] The dimension of the pads and the surrounding grid is such that either (i) objects, such as a finger, are inhibited from contacting the array 16 because the grid functions as a barrier or (ii) if the array 16 is contacted by an object it first contacts the grid. Therefore, any charge accumulated on a user's hand, or tool they are holding, is inhibited from passing through the well 28 or well electrode 26 region if it approaches or contacts the component 8 in the region of the array of electrodes 16 because it is directed through or via the grid 102 to the common electrode 36.
[0083] Because the grid 102 and polar medium 32 are electrically connected, energy transferred from an ESD to the grid and to the polar medium requires negligible work done such that the voltage across a well 28 is negligible. In use, the layer of polar medium 32, which can contain a sample to be analysed, is electrically connected to the common electrode 36 via the conductive diffusion layer 40. The grid 102, common pad 104, common electrode 36, conductive diffusion layer 40 and polar medium 32 or sample have, therefore, negligible difference in potential between them if an ESD was applied to the grid. This is because the charge is distributed across these components, which inhibits any charge passing between an electrode pad 100 and the well 28 to the sample of polar solution when the grid is contacted or both the grid and a pad are contacted. Sensors and/or membranes formed or forming part of the well 28 are protected from an ESD to the array 16. The grid 102 acts as a shield—mechanically and/or electrically—that is connectable to a substantially large volume of fluid i.e. the polar medium 32. These elements of the component 8 are significantly larger, by at least 2 orders of magnitude, than the size of the wells 28 or the volume of fluid held in the wells. As described above, the ratio of the volume of fluid in a well 28 to the volume of the fluid in the layer above that forms the polar medium 32 can be between about 1:100 about 1:10000. In the example, the common cell chamber, or sample chamber for holding a polar medium 32 and associated common mediator chamber, has a volume of about 135 ul. In the example, which has an array of 130 electrodes, 126 are occupied by fluid for forming a membrane between each well and the sample chamber above and the total volume of fluid in the wells is about 0.9 ul. The ratio in the example is about 1:150. In this way, the grid and/or the volume of polar medium (which is significantly larger than all of the wells together) function as a buffer or insulator protecting the sensing elements of the wells and sensor elements therein from an ESD or similar uncontrolled charge. In other words, any uncontrolled charge or ESD is inhibited from conducting from a pad 100 to a well 28 because the grid 102 inhibits contact with the pad and/or inhibits contact with the pad without also contacting the grid such that either (i) a charge, such as an ESD, passes to the polar medium or sample to distribute the charge with such low energy consumption that there is negligible potential difference between the pads 100 and the well 28 or (ii) if the array 16 is contacted an ESD charge is inhibited from flowing across the well 28 because the grid 102 has been contacted such that the well 28 region and the grid have the same potential difference—the charge is already balanced.
[0084] In use, the component is packaged in material that inhibits the build up of charge to minimise the risk of damage to a sensor or membrane within a well 28, which can occur if a pad 100 of the array of electrodes 16 is touched. In field use, outside of a controlled environment such as a laboratory, there are rarely facilities such as earth-points or earth straps to divert uncontrolled voltages or ESD away from the pads.
[0085] To illustrate the dimensions of the invention, by way of an example illustration, the tip of a finger 106 is shown adjacent the grid 102 in
[0086] After initial manufacture of the component 8 and before population with a conductive fluid, such as a polar medium 32, the grid 102 is configured to protect any sensor that will subsequently be formed in the well 28.
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[0089] A gap is provided between the pad 100 and the grid 102. The grid has been illustrated in other Figures as an extension of the substrate, preferably grown from the substrate, upon which the pad 100 is formed, which cannot be appreciated from the plan view of
[0090] Alternatively, the grid can lie flush with the surface of the array such that finger-contact with a pad is not inhibited but finger-contact with a pad 100 without touching the grid is inhibited.
[0091] The array 16 can additionally be covered by a protective antistatic tape (not shown) that can be removed from the array 16 on the component 8 prior to insertion and connection with the receiver 6. Without the grid 102 such tape could generate a triboelectric charge when peeled from the array 16 and damage the sensing function in a well 28. The tape, however, complements the function of the grid 102 because any triboelectric charge generated from its removal will not influence the sensing in the well because the tape is connected to the grid. The conductive structure is therefore covered, at least in part, with a removable protective conductive film or protective layer.
[0092] Mechanical connections, such as spring-leaf metal contacts located on the receiver 6 for engagement with the electrodes on the component 8 can be provided.
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[0094] The height of a grid above the substrate—whether in net form, or whether in the form of walls as shown in
[0095] As described above, the shape and formation of the grid is such that the inductance between the grid 102, common pad 104 and common electrode 32 is minimised.
[0096] The array of electrodes 16 and/or grid 102 have been described as formed on a substrate 24 with conductive surfaces for the pads 100 and for connection to, for example, the common electrode 36.
[0097] The purpose of the component is to provide a low cost single use device and in light of the teaching herein various low-cost manufacturing techniques are applicable to the examples.
[0098] Alternatively, the pads 100 and grid 102 or tracks can be formed on, or by using:
[0099] silicon wafer fabrication, which can provide a grid minimum width as small as 5 um, with a gap between the pad and grid of 5 um;
[0100] glass wafer fabrication, which can provide a grid minimum width as small as 10 um, with a gap between the pad and grid of 10 um;
[0101] printed circuit boards, which can provide a grid minimum width as small as 50 um, with a gap between the pad and grid of 50 um;
[0102] high resolution printed circuit boards, which can provide a grid minimum width as small as 3 um, with a gap between the pad and grid of 5 um;
[0103] conductive ink, inkjet printed on card or polymers, which can provide a grid minimum width as small as 150 um, with a gap between the pad and grid of 50 um;
[0104] screen printing conductive ink on card or polymers, which can provide a grid minimum width as small as 20 um, with a gap between the pad and grid of 20 um; and
[0105] sputtering conductive material on card or polymers, which can provide a grid minimum width as small as 10 um, with a gap between the pad and grid of 10 um.
[0106] By way of example, an inkjet printed, roll-coated digital microfluidic device for inexpensive, miniaturized diagnostic assays is known from a paper in the name of Dixon et al [Lab Chip, 2016,16, 4560].
[0107] In
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[0109] Like numerals in the Figures represent like features. The present invention has been described above purely by way of example, and modifications can be made within the spirit and scope of the invention, which extends to equivalents of the features described and combinations of one or more features described herein. The invention also consists in any individual features described or implicit herein.
TABLE-US-00001 List of features: 2 Apparatus 4 Device 6 Receiver 8 Component 10 Base 12 Latch 14 Recess 16 Array of electrodes 18 Array of connectors 20 Solder bump 22 Via 24 Substrate 26 Well electrode 28 Well/Recess 30 Seat 32 Layer of polar medium 34 Meniscus/membrane 34a Membrane 36 Common electrode 40 Conductive diffusion layer 100 Pad 102 Grid/conductive structure 104 Common pad 106 Finger tip 108 Connectors 110 Recess 112 Track