Multilayer electrical connection for digital microfluidics on substrates

Abstract

An apparatus for forming a plurality of microdroplets from a droplet includes a substrate, a dielectric layer on the substrate and having a plurality of hydrophilic surface regions spaced apart from each other by a hydrophobic surface, and a plurality of electrodes covered by the dielectric layer. The electrodes are configured to form an electric field across the droplet in response to a voltage provided by a control circuit to move the droplet across the dielectric layer in a lateral direction while leaving a portion of the droplet on the hydrophilic surface regions to form a plurality of microdroplets on the hydrophilic surface regions.

Description

[0001] This application is a divisional application of patent application No. 201980073257.0, filed on November 8, 2019, by Shenzhen BGI Genomics Co., Ltd., entitled "Multilayer Electrical Connection of Digital Microfluidics on Substrate".

[0002] Cross-reference to related applications

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 758,071, filed November 9, 2018, which is incorporated herein by reference. Technical Field

[0004] Embodiments of the present invention generally relate to electrowetting on dielectric (EWOD) technology, and more specifically to apparatus and methods for manipulating droplets using a multilayer ceramic substrate, disposable substrates operable with the multilayer ceramic substrate, and disposable substrates with sockets for integrated circuit packaging. Background Technology

[0005] Electrowetting on dielectric (EWOD) is a liquid-driven mechanism used to change the contact angle of a water droplet between two electrodes on a hydrophobic surface. Bulk droplets as large as a few millimeters (i.e., a few microliters in volume) can be moved by an array of electrodes disposed on a substrate (e.g., an inorganic substrate, such as a silicon / glass substrate) or an organic substrate (e.g., a cyclic olefin polymer / polycarbonate substrate).

[0006] Figure 1A This is a perspective view illustrating an EWOD device 10 that can be used to explain embodiments of the present disclosure. The EWOD device includes a substrate structure 11 having a substrate 12, an insulating layer 13 on the substrate, and an electrode array 14 within or beneath the insulating layer. The electrode array 14 includes a first set of electrodes 14a arranged parallel to each other and spaced apart from each other in a first direction, and a second set of electrodes arranged parallel to each other and spaced apart from each other in a second direction substantially perpendicular to the first direction. The first and second sets of electrodes are spaced apart from each other within the insulating layer 13, which may include multiple dielectric layers having the same or different materials. The EWOD device also includes an input-output circuit 15 in the substrate, which interfaces with control circuitry that can be integrated into or external to the EWOD device to provide a control voltage having a time-varying voltage waveform to the electrode array 14.

[0007] Reference Figure 1A By turning off / off the control voltage at the electrode below the droplet and the adjacent electrode, the droplet 16 arranged on the surface of the insulating layer 13 can be moved in a specific direction.

[0008] Figure 1Byes Figure 1A The EWOD device shown is a cross-sectional view taken along line B-B'. A cross-sectional view of the second set of electrodes 14b is shown. The first set of electrodes 14a (not shown) may be disposed above, below, or in the same plane as the second set of electrodes 14b, and spaced apart from the second set of electrodes by one or more dielectric layers. Summary of the Invention

[0009] Embodiments of this disclosure provide apparatus, systems, and methods for manipulating droplets containing reagents to obtain mixed droplets, forming a large number of tiny droplets (particles, microdroplets, or samples) from the mixed droplets, reading the DNA concentration of the sample (microdroplet) by optical detection, or measuring the pH of each sample by an integrated ion-sensitive field-effect transistor (ISFET) sensor, thereby calculating the DNA concentration of the droplets. It should be noted that although the embodiments describe apparatus and processes for measuring the pH of droplets, this disclosure is not limited thereto. Those skilled in the art will understand that, based on pH changes within the tiny droplets (microdroplets), the apparatus and methods described herein can be applied to pH measurements of any aqueous and non-aqueous liquids.

[0010] In one embodiment of this disclosure, a device for manipulating droplets may include: a ceramic substrate having a first surface and a second surface; a first electrode array disposed on the first surface; a plurality of contact pads spaced apart from each other and disposed on the second surface; and one or more interconnect layers disposed between the first surface and the second surface and configured to electrically couple one or more of the plurality of contact pads to one of the first electrode array. In some embodiments, the ceramic substrate is a multilayer ceramic substrate comprising an inorganic dielectric layer (e.g., silicon dioxide, silicon nitride) on the first surface and a hydrophobic layer on the inorganic dielectric layer. In other embodiments, the ceramic substrate is a multilayer ceramic substrate comprising an organic dielectric layer (e.g., polyimide) on the first surface and a hydrophobic layer on the inorganic dielectric layer.

[0011] In some embodiments, the device may further include: a monolayer substrate having a third surface and a fourth surface and a plurality of through-holes extending from the third surface to the fourth surface; a second electrode array on the third surface of the ceramic substrate; and a plurality of conductive features coupled to the second electrode array and filling the through-holes. Each of the conductive features may include a protrusion protruding from the fourth surface of the monolayer substrate and aligned with one of the first electrode arrays. The through-holes are aligned with the first electrode array. In some embodiments, the monolayer substrate is a disposable substrate and may include glass, ceramic, organic materials, plastics, etc.

[0012] In another embodiment, a device for manipulating droplets may include: a socket for supporting an integrated circuit package, comprising: a plurality of contact pads disposed on a flat surface of the socket and configured to receive electrical signals from the integrated circuit; a monolayer substrate having a first surface, a second surface, and a plurality of through-holes extending through the monolayer substrate; and a plurality of electrodes on the first surface of the monolayer substrate, electrically coupled to the plurality of contact pads through the through-holes, and configured to manipulate the droplets disposed on the first surface of the monolayer substrate by means of the electrical signals provided by the integrated circuit. In some embodiments, the socket is a ball grid array socket. In other embodiments, the socket is a pad grid array socket. In some embodiments, the monolayer substrate is a disposable substrate and may comprise glass, ceramic, organic materials, or plastics.

[0013] In another aspect, an apparatus for manipulating droplets may include: a multilayer ceramic substrate having a first surface and a second surface, the multilayer ceramic substrate including a plurality of electrically insulating layers and a plurality of electrically conductive layers (metallic wires) between the electrically insulating layers; a plurality of driving electrodes on the first surface and configured to perform a plurality of manipulation operations on the droplets; a plurality of contact pads on the second surface and electrically coupled to one or more of the driving electrodes to provide a reference voltage; a dielectric layer covering the first surface including the driving electrodes; and a hydrophobic layer on the dielectric layer. In some embodiments, each of the driving electrodes has a size smaller than the area occupied by the droplets. In some embodiments, the plurality of manipulation operations include: conveying the droplets along a travel path on the first surface; mixing the droplets with one or more reagents to obtain mixed droplets; splitting the mixed droplets into a plurality of microdroplets for detection, and collecting the microdroplets in a waste area after the detection. In some embodiments, the first surface of the multilayer ceramic substrate includes: a first region having an inlet for receiving the droplet; a second region having one or more reservoirs for storing one or more reagents; a third region communicating with the first and second regions and configured to mix the droplet with the one or more reagents to obtain a mixed droplet; a fourth region communicating with the third region and configured to separate the mixed droplet into a plurality of equally sized microdroplets for analysis; and a fifth region communicating with the fourth region and configured to collect the microdroplets after analysis. In some embodiments, the device further includes a second substrate opposite to and forming, together with the multilayer ceramic substrate, a channel for the droplet and the microdroplets, the second substrate including a sixth region facing the fourth region, the sixth region including a plurality of sensors associated with the microdroplets. In some embodiments, each of the sensors may include: an ion-sensitive field-effect transistor including an ion-sensing membrane configured to be exposed to a solution contained in the droplets and to provide a signal relating to the concentration level of the droplets in the solution; and a reference voltage electrode configured to apply a reference voltage to the solution.

[0014] In another aspect, a method of manufacturing a device for manipulating droplets may include: providing a multilayer ceramic substrate having a first surface and a second surface, the first surface including an array of driving electrodes, the second surface opposite the first surface and including a plurality of contact pads electrically contacting one or more driving electrodes in the array of driving electrodes; coating a dielectric layer on the first surface of the multilayer ceramic substrate; and coating the hydrophobic layer on the dielectric layer. In some embodiments, the method may further include: providing a second substrate including a third surface, a fourth surface opposite the third surface, and a plurality of through-holes extending through the second substrate. The second substrate includes an array of electrode pads disposed on the third surface and a plurality of conductive features filling the through-holes. Each of the conductive features includes a protrusion protruding from the fourth surface and aligned with one of the driving electrode arrays. In some embodiments, the method may further include: forming a hydrophobic dielectric layer on the third surface of the second substrate. In some embodiments, the method may further include: attaching the second substrate to the multilayer ceramic substrate using a fastening member such that the array of electrode pads is electrically and physically in contact with the array of driving electrodes.

[0015] In another aspect, a method of manufacturing an apparatus for manipulating droplets may include: providing a socket for supporting a package containing an integrated circuit, the socket including a plurality of contact pads disposed on a flat surface of the socket and configured to receive electrical signals from the integrated circuit; providing a monolayer substrate having a first surface, a second surface and a plurality of through-holes extending through the monolayer substrate, and a plurality of electrodes on the first surface of the monolayer substrate and electrically coupled to the plurality of contact pads through the through-holes, and configured to manipulate the droplets disposed on the first surface of the monolayer substrate by means of the electrical signals provided by the integrated circuit.

[0016] In another aspect, a device for manipulating a droplet may include: a ceramic substrate having a first surface for receiving the droplet; a dielectric layer disposed on the first surface of the ceramic substrate; and a plurality of electrodes covered by the dielectric layer and configured to manipulate the droplet in response to an electrical signal provided to the plurality of electrodes. In one embodiment, the device may further include a opposing substrate having a second surface facing the first surface of the ceramic substrate and spaced apart from the ceramic substrate to form an air gap for the droplet. The opposing substrate includes a common electrode disposed on the second surface. The opposing substrate may also include a plurality of sensors configured to detect characteristics of the droplet.

[0017] This summary is provided to illustrate different embodiments of the present disclosure in a simplified form, which will be described in detail below. This summary is not intended to limit the scope of the claimed subject matter. Other features, details, utility, and advantages of the claimed subject matter will become apparent from the following detailed description.

[0018] definition

[0019] The terms "wafer" and "substrate" should be understood to include silicon-on-insulator (SOI) or silicon-on-sapphire (SOS) technologies, doped and undoped semiconductors, epitaxial layers of silicon supported by a substrate semiconductor base, and other semiconductor structures. Furthermore, when "wafer" or "substrate" is mentioned in the following description, prior processes may have been used to form regions or junctions within the substrate semiconductor structure or base. Additionally, the semiconductor need not be silicon-based, but can be based on silicon-germanium, germanium, or gallium arsenide.

[0020] As used herein, the term "computer-readable medium" refers to any medium that participates in providing instructions to the processor of a controller for execution. Computer-readable media can take many forms, including but not limited to non-volatile media, volatile media, and transmission media. Non-volatile media include, for example, optical discs, magnetic disks, and magneto-optical disks, such as hard disks or removable media drives. Volatile media include dynamic memory, such as main memory. Furthermore, various forms of computer-readable media may be involved when the processor of the controller executes one or more sequences of one or more instructions. For example, instructions may initially be carried on a disk of a remote computer. The remote computer may remotely load all or part of the instructions used to implement the invention into dynamic memory and send the instructions to the controller via a network.

[0021] The term "hydrophobic" refers to a material that has a contact angle with water in air greater than or equal to 90 degrees. In some embodiments, a hydrophobic surface may have a contact angle greater than 90 degrees, such as 120 degrees, 150 degrees, etc. Conversely, the term "hydrophilic" refers to a material that has a contact angle with water in air or an immiscible liquid such as oil less than 90 degrees.

[0022] The term "droplet" has its normal meaning in this technical field and refers to a droplet with a certain volume (e.g., about a few milliliters) having a boundary at least partially formed by surface tension. -3 ) to about a few microliters (10 -6A droplet is a liquid. Droplets can be water-based (aqueous) droplets, which include any organic or inorganic substance, such as biomolecules, proteins, living or dead organisms, reagents, and any combination thereof. Droplets can also be non-aqueous liquids. Droplets can be spherical or non-spherical, and their size ranges from 1 micrometer to approximately several millimeters. Droplets can be divided into multiple very small portions (small droplets) spaced apart from each other and having a substantially uniform size. The volume of a very small portion can be as small as 1 microliter (10⁻⁶). -6 L or mL) and 100 nanoliters (10 -9 Between 100 nL and 10 nL, and between 10 nL and 100 pL (10 nL or nL) -12 The volume ranges from 100 pL to between 10 pL. In some embodiments, very small fractions can have a volume of several picoliters. In this disclosure, very small fractions are alternatively referred to as microdroplets.

[0023] The term "reagent" refers to a molecule or compound of different molecules that can induce a specific reaction with substances present in a droplet. Attached Figure Description

[0024] Figure 1A This is a simplified perspective view illustrating an EWOD device that can be used to explain embodiments of this disclosure.

[0025] Figure 1B yes Figure 1A The simplified cross-sectional view of the EWOD device shown is taken along line B-B'.

[0026] Figure 2A This is a simplified cross-sectional view of a part of an EWOD device according to an embodiment of this disclosure.

[0027] Figure 2B This is a simplified cross-sectional view of a part of an EWOD device according to another embodiment of this disclosure.

[0028] Figure 2C This is a simplified cross-sectional view of a part of an EWOD device according to yet another embodiment of this disclosure.

[0029] Figure 2D This is a cross-sectional view of a portion of an EWOD device having a second substrate according to another embodiment of the present disclosure, the second substrate including a plurality of sensors.

[0030] Figure 2E This is a cross-sectional view of a portion of an EWOD device having a second substrate according to another embodiment of the present disclosure, the second substrate including a plurality of sensors and one or more reference electrodes.

[0031] Figure 2FThis is a simplified plan view of an EWOD device according to an exemplary embodiment of the present disclosure.

[0032] Figure 2G This is a simplified cross-sectional view showing the electric field generated by the electrodes according to an embodiment of the present disclosure.

[0033] Figure 2H This is a simplified cross-sectional view showing the electric field generated by the electrodes according to another embodiment of the present disclosure.

[0034] Figure 2I This is a simplified cross-sectional view showing the electric field generated by the electrodes according to yet another embodiment of the present disclosure.

[0035] Figures 3A to 3C This is a simplified top view of a droplet moving on the surface of a dielectric layer according to an embodiment of the present disclosure. Figure 3A This is a simplified top view showing the discharge of droplets on the first electrode of the electrode array according to an embodiment of the present disclosure. Figure 3B This is a simplified top view illustrating, according to an embodiment of the present disclosure, the movement of a droplet to a second (adjacent) electrode under the influence of an electric field via an EWOD device. Figure 3C This is a simplified top view illustrating, according to an embodiment of the present disclosure, the removal of a droplet from the electrode array while leaving a residue on the second electrode.

[0036] Figure 4A This is a cross-sectional view of a portion of an EWOD device according to an embodiment of this disclosure.

[0037] Figure 4B This is a cross-sectional view of a portion of an EWOD device according to another embodiment of this disclosure.

[0038] Figure 4C yes Figure 4A A perspective view of a portion of the EWOD device shown.

[0039] Figure 4D yes Figure 4B A perspective view of a portion of the EWOD device shown.

[0040] Figure 4E This is a perspective view of a portion of an EWOD device according to an embodiment of this disclosure.

[0041] Figure 5 This is a cross-sectional view of an ISFET device according to an embodiment of the present disclosure.

[0042] Figure 6A This is a simplified top view of an integrated on-chip laboratory device according to an embodiment of this disclosure.

[0043] Figure 6BThis is a simplified top view of an exemplary arrangement of electrodes in an integrated on-chip laboratory device according to an embodiment of this disclosure.

[0044] Figure 7A This is a simplified cross-sectional view of a multilayer ceramic substrate that can be used as an EWOD, according to an embodiment of the present disclosure.

[0045] Figure 7B This is a simplified cross-sectional view showing a multilayer ceramic substrate configured as an insert substrate for use as a single-layer substrate according to an embodiment of the present disclosure.

[0046] Figure 7C This is a perspective view of an exemplary single-layer substrate according to an embodiment of the present disclosure.

[0047] Figure 7D This is a simplified cross-sectional view of a multilayer ceramic substrate coupled to a single-layer substrate via a fastening member, according to an embodiment of this disclosure.

[0048] Figure 8A This is a simplified cross-sectional view of a socket for supporting an integrated circuit in accordance with an embodiment of this disclosure.

[0049] Figure 8B A ball grid array socket according to an embodiment of the present disclosure is shown, which can be used as an intermediary layer (interconnect) between test instruments and a disposable substrate.

[0050] Figure 8C A pad grid array socket, which can be used as an intermediary layer (interconnect) between a test instrument and a disposable substrate, is shown according to an embodiment of the present disclosure.

[0051] Figure 8D A simplified cross-sectional view of a single-layer substrate including multiple sensors is shown according to an embodiment of the present disclosure.

[0052] Figure 9 This is a simplified flowchart illustrating a method for manufacturing an apparatus for manipulating droplets according to an embodiment of the present disclosure.

[0053] Figure 10 This is a simplified flowchart illustrating a method for manufacturing an apparatus for manipulating droplets according to an embodiment of the present disclosure.

[0054] Figure 11 This is a simplified schematic diagram of a mobile computing device according to an embodiment of the present disclosure, which can be used to control a reference. Figure 1A , 1B The apparatus or EWOD device described in 2A-2I, 3A-C, 4A-B, 5, 6A-B, 7A-D, 8A-C, 9 and 10.

[0055] Figure 12This is a cross-sectional view illustrating the process of manufacturing a multilayer ceramic substrate structure according to an embodiment of this disclosure.

[0056] Figures 13A to 13H A cross-sectional view is shown illustrating an intermediate step in the process of manufacturing a single-layer substrate according to an embodiment of the present disclosure.

[0057] As is customary, the features and elements described are not drawn to scale, but are drawn to emphasize the features and elements relevant to this disclosure. Detailed Implementation

[0058] It should be understood that, for the sake of simplicity and clarity, the elements shown in the figures are not necessarily drawn to scale. For example, for clarity, the dimensions of some elements are enlarged relative to each other. Furthermore, reference numerals are repeated between figures where deemed appropriate to indicate corresponding elements.

[0059] In the following detailed description, reference is made to the accompanying drawings, which form a part thereof, and specific embodiments in which the invention may be implemented are illustrated by way of illustration. The orientations in the described drawings are referred to using the terms “upper,” “lower,” “vertical,” “horizontal,” “depth,” “height,” “width,” “top,” “bottom,” etc. Because components of embodiments of the invention can be positioned in many different orientations, these terms are used for illustrative purposes and not for limitation.

[0060] The terms "first," "second," etc., used do not indicate any order, but are used to distinguish one element from another. Furthermore, the use of terms "a," "an," etc., does not indicate a limitation on quantity, but rather indicates the presence of at least one of the referenced items.

[0061] As used herein, turning off an electrode means reducing the voltage of that electrode to a level below the common voltage, such as connecting the electrode to ground potential. Alternatively, turning off an electrode may also mean setting the electrode to a floating state. Conversely, turning on an actuator electrode means increasing the voltage of that actuator electrode to a level above the common voltage. The common voltage can be any voltage shared by multiple circuit elements of the EWOD device, such as ground potential.

[0062] As used herein, a droplet is an encapsulated liquid. Droplets can be spherical or non-spherical. A droplet can be divided into multiple very small portions (microdroplets) that are separated from each other and have a substantially uniform size. In this disclosure, the very small portions of a droplet may alternatively be referred to as microdroplets.

[0063] Figure 2A This is a simplified cross-sectional view of a portion of an EWOD device 20A according to an embodiment of this disclosure. (Refer to...) Figure 2AThe EWOD device 20A includes a first substrate 22, a dielectric layer 23 on substrate 21, a group of actuating electrodes 24 (e.g., 24a, 24b, 24c) within the dielectric layer 23, and a common electrode 27 attached to a second substrate 28 and facing the actuating electrodes 24. The common electrode 27 may be grounded or have other common voltages. The dielectric layer 23 and the common electrode 27 are spaced apart from each other by spacers 29. (Reference) Figure 2A A droplet 26 is disposed between an actuating electrode 24 and a common electrode 27, and moves laterally across the surface of the dielectric layer 23 by changing or altering the voltage level applied to the actuating electrode relative to the common electrode. In one embodiment, the EWOD device 20A may further include control circuitry (not shown) configured to provide control voltages to the common electrode and the actuating electrode. By switching the voltage applied to the actuating electrode on and off, the control circuitry can cause the droplet 26 to move laterally across the surface of the dielectric layer 23. For example, an electric field is generated by applying a first voltage to the actuating electrode 24a below the droplet 26 and a second voltage to the adjacent actuating electrode 24b, causing the droplet 26 to move toward the actuating electrode 24b. The speed of movement of the droplet 26 can be controlled by the magnitude of the voltage difference between adjacent actuating electrodes. In one embodiment, when the droplet 26 is disposed between the actuating electrode 24 and the common electrode 27, the shape of the droplet 26 can be changed by altering the voltage difference between the actuating electrode 24 and the common electrode 27. It should be understood that the number of actuators in a group of actuator electrodes can be any integer. Figure 2A In the example shown, three actuation electrodes are used in the grouped actuation electrodes. However, it should be understood that this number is arbitrarily chosen to describe the example implementation and should not be limiting.

[0064] Reference Figure 2A Two substrate structures can be formed separately. For example, a first substrate structure can be formed, which includes a substrate 22, a dielectric layer 23, and an actuation electrode 24 within the dielectric layer 23. The substrate 22 can be a thin-film transistor array substrate formed by conventional thin-film transistor (TFT) manufacturing processes. The second substrate structure can include a substrate 28 and a common electrode 27 layer on the substrate 28. Spacers 29 can be formed on either the first or second substrate structure. In some embodiments, the height of the spacers 29 is in the range of several micrometers to several millimeters. Typically, the height of the spacers 29 is smaller than the diameter of the droplet, such that the droplet disposed on the dielectric layer 23 is in physical contact with the second substrate structure. The first and second substrate structures are then combined to form an EWOD device 20A. In other words, the space or air gap between the first and second substrate structures is determined by the height or thickness of the spacers 29. This space or air gap forms a channel for the droplet.

[0065] exist Figure 2A In the illustrated embodiment, the common electrode 27 and the group of actuating electrodes 24 (e.g., 24a, 24b, 24c) are connected via... Figure 1A The input-output circuit 15 shown is connected to a voltage provided by a control circuit (not shown). In some embodiments, the common electrode may be connected to ground potential or a stable DC voltage. The control circuit applies a time-varying voltage to the group of actuating electrodes via the input-output circuit through a corresponding electronic switch (which may be, for example, a thin-film transistor or MOS circuit outside the substrate or chip) to generate an electric field across the droplet, thereby causing the droplet to move along a path. In some embodiments, the surface of the common electrode 27 is covered with an insulating layer made of a hydrophobic material. In other embodiments, the surface of the dielectric layer 23 is coated with a hydrophobic film having a submicron thickness.

[0066] Figure 2B This is a simplified cross-sectional view of a portion of an EWOD device 20B according to another embodiment of this disclosure. Reference Figure 2B The EWOD device 20B includes a substrate 22b, a dielectric layer 23b on the substrate 21b, a group of actuating electrodes 24 (24a, 24b, 24c) within the dielectric layer 23b, and a group of common electrodes 27 (only one electrode 27b is shown) covering the dielectric layer 23b. The common electrode 27b and the actuating electrodes are spaced apart from each other by a portion of the dielectric layer. Similar to... Figure 2A By applying a first voltage at an actuation electrode (e.g., 24a) below the droplet 26 and a second voltage at an adjacent actuation electrode (e.g., 24b), the droplet 26 can move along a lateral path across the surface of the dielectric layer 23b. The movement and orientation of the droplet 26 are thus controlled by a control circuit (not shown) that applies voltages to certain actuation electrodes via a group of electronic switches (MOS circuitry in substrate 22b, not shown). Unlike the EWOD 20A shown in FIG. 20A, the EWOD device 20B has a common electrode 27b close to the actuation electrode 24, and the droplet 26 is not sandwiched between the common electrode 27b and the actuation electrode 24. The EWOD device 20B also differs from the EWOD 20A in that it lacks the spacer 29.

[0067] Reference Figure 2B The actuating electrodes 24 and the common electrodes 27 can be two strip electrodes intersecting each other on different planes on the substrate. The actuating electrodes 24 and the common electrodes 27 are operable to move the droplet 26 across the surface of the dielectric layer 23b. In some embodiments, the common electrode 27b has a surface covered by an insulating layer made of a hydrophobic material. In other embodiments, the surface of the dielectric layer 23 is coated with a submicron-thick hydrophobic film.

[0068] Figure 2CThis is a cross-sectional view of a portion of an EWOD device 20C according to yet another embodiment of this disclosure. Reference Figure 2C The EWOD device 20C includes a substrate 22c, a dielectric layer 23c on the substrate 22c, a group of actuating electrodes 24 (e.g., 24a, 24b, 24c) within the dielectric layer 23c, and a group of common electrodes (e.g., a common electrode 27c shown) covering the dielectric layer 23c. The common electrode 27c and the actuating electrodes are spaced apart from each other by a portion of the dielectric layer. In some embodiments, the common electrode 27c has a surface covered by an insulating layer made of a hydrophobic material or covered by a submicron hydrophobic coating film on the surface of the dielectric layer 23. The EWOD device 20C may also include a second substrate 28c spaced apart from the substrate 21c by spacers 29c. Similar to Figure 2A The droplet 26 can move along a path within a channel formed by the space or air gap between the surface of the dielectric layer and the second substrate 28c. The movement of the droplet is controlled by a control circuit (not shown) through a voltage applied to the electrode via an electronic switch.

[0069] Figure 2D This is a cross-sectional view of a portion of an EWOD device 20D according to yet another embodiment of this disclosure. Figure 2D Similar to Figure 2C The difference is that the second substrate 28d includes multiple sensors configured to detect the characteristics of the droplets.

[0070] Figure 2E This is a cross-sectional view of a portion of an EWOD device 20E according to yet another embodiment of this disclosure. Figure 2E Similar to Figure 2D The difference is that the common electrode 27e is integrated in the second substrate 28e together with multiple sensors configured to detect the characteristics of the droplet.

[0071] Figure 2F This is a simplified plan view of an EWOD 20D according to an exemplary embodiment of this disclosure. Reference Figure 2FThe actuating electrodes are arranged in an array having routing channels for routing electrical signals from control circuitry 28 to actuating electrode 24 and common electrode 27c. The spacers 29c shown have a circular cross-section; however, the circular cross-sectional shape is not limiting, and any other cross-sectional shape is equally applicable, such as square, rectangular, oval, elliptical, and other shapes. Similarly, the actuating electrodes are shown as having a square shape; however, the square shape is not limiting, and other shapes are equally applicable, such as rectangular, circular, oval, elliptical, and other shapes. In one embodiment, the spacers 29c are spaced apart to leave sufficient space to allow droplet free movement. In other words, the size and spacing of the spacers 29c are set such that they do not impede the movement of droplets across the entire dielectric layer surface. It should be understood that although the routing channels are shown as being coplanar with the electrode array, those skilled in the art will understand that the routing channels and control circuitry can be disposed in the substrate and in different layers of the dielectric layer. It should also be understood that the actuating electrodes 24 and common electrode 27c can have their relative positions transposed, i.e., the common electrode can be disposed below the actuating electrodes.

[0072] In another embodiment, the EWOD device may have a single electrode array. In other words, the common electrode and the actuating electrode are coplanar, meaning they are arranged in the same plane within the dielectric layer. For example, multiple actuating electrodes and multiple common electrodes are arranged alternately adjacent to each other, and the control circuitry can sequentially apply DC or AC voltage and ground potential to the actuating electrodes and common electrodes to control the movement of the droplet. In yet another embodiment, each electrode in the electrode array is individually controlled by the control circuitry via a group of electronic switches, such that each electrode can be an actuating electrode in a first time period and a common electrode in a second time period.

[0073] Figure 2G This is a simplified cross-sectional view illustrating an exemplary electric field E generated by electrodes according to an embodiment of the present disclosure. Reference Figure 2G The common electrode 27 can be driven by ground potential (gnd), and the actuating electrodes 24a, 24b, and 24c can be driven sequentially by DC or AC voltage (Ve). For example, first, a DC voltage is applied to electrode 24a, and the voltage difference between actuating electrode 24a and common electrode 27 generates an electric field E, which causes droplet 26 to move along the microchannel defined by electrode 27 and dielectric layer 23. By setting a voltage at an electrode adjacent to droplet 26, droplet 26 can be moved to that electrode in the lateral direction between dielectric layer 23 and common electrode 27. This structure is similar to... Figure 2A The EWOD device shown is similar to or the same. In one embodiment, the DC voltage can be controlled by the control circuit. Figure 1A The input-output circuit 15 shown is provided. The main electric field is perpendicular to the surface of the dielectric layer 23.

[0074] Figure 2H This is a simplified cross-sectional view illustrating an exemplary electric field generated by electrodes according to another embodiment of this disclosure. Reference Figure 2H A control circuit (not shown) can apply DC or AC voltages to electrodes 24a, 24b, 24c, and 27b in a time-sequential manner to generate an electric field pattern that controls the movement of droplet 26. For example, a voltage applied to an electrode adjacent to the droplet will move the droplet onto that electrode. This structure is similar to... Figure 2B Or the EWOD device shown in 2C is the same as or similar to it.

[0075] Figure 2I This is a simplified cross-sectional view showing an electric field 29I generated by electrodes according to another embodiment of this disclosure. (See reference) Figure 2I A control circuit (not shown) can apply DC or AC voltage to electrodes 24a, 24b, and 24c in a time-sequential manner to generate an electric field pattern that controls the movement of droplet 26. In this embodiment, electrode patterns 24a, 24b, and 24c can alternatively serve as actuating electrodes and a common electrode. A semi-cylindrical field is formed between electrodes 24a and 24b. (Reference) Figure 2G , 2H And 2I, the electric field E and the generated electric force F are functions of the voltage difference between the electrodes and the electrode size. By changing the voltage difference between adjacent electrodes in a time-sequential manner, an electric field and a resultant force are generated, thereby causing the droplet 26 to be transported along the electric direction. In some embodiments, non-active electrodes (e.g., electrode 24c) that do not contribute to the movement (or transport) of the droplet can remain floating, i.e., not connected. Figure 2I In the example shown, droplet 26 will remain between electrodes 24a and 24b, that is, between voltage Ve and ground.

[0076] Figures 3A to 3C This is a top-sequence view of a droplet moving across the surface of the entire dielectric layer according to an embodiment of this disclosure. Reference Figure 3ADroplet 26 is disposed on an EWOD device, as described above in any of EWOD devices 20A, 20B, and 20C. The EWOD device includes a substrate with a thin-film transistor array or MOS circuitry, a dielectric layer on the substrate, and an array of actuating electrodes (and / or common electrodes) within the dielectric layer. The actuating electrodes and common electrodes are connected to control circuitry via wires in a routing channel and receive control signals from the control circuitry via thin-film transistors. Droplet 26 is disposed on the surface of the dielectric layer above a first actuating electrode 34a. By closing (or floating) the first driving electrode below the droplet and opening the next driving electrode adjacent to it, the droplet can move toward the next electrode. In one embodiment, a predetermined feature can be used to modify a portion of the surface of the dielectric layer above the actuating electrode array, which has a greater attraction to liquids (e.g., droplets) than a hydrophobic surface. This feature can have dimensions ranging from micrometers to nanometers, corresponding to microliters and nanoliters, respectively. Feature 35 can be precisely fabricated on the dielectric layer thousands or millions of times using currently available submicron semiconductor manufacturing processes.

[0077] As used herein, turning off an actuating electrode means reducing the voltage of the actuating electrode to the same level as the common voltage applied to the common electrode. Conversely, turning on an actuating electrode means increasing the voltage of the actuating electrode to a level higher than the common voltage. The EWOD device can operate at DC (DC electrowetting) or AC (AC electrowetting) voltages, provided that the potential between the electrodes is at a DC voltage level to create an electric field for moving the droplet. In some embodiments, when an adjacent electrode is fully or partially turned on, a droplet positioned adjacent to it will be moved to the turned-on electrode and wet a feature placed on the turned-on electrode. As used herein, the term "feature" refers to an area or structure in which a liquid material (e.g., a droplet) is deposited or formed. By moving the droplet to the next turned-on electrode using a time-varying voltage waveform provided by control circuitry, the droplet will move between the electrodes, leaving residual microdroplets (very small or minute drops or microparticles)26a in or on the feature. The volume of residual microdroplets depends entirely on the characteristic size and the contact angle of the liquid droplets on the surface of the environment (e.g., air or oil). Figure 3B This is a top view showing a droplet 26 moving from a first electrode 34a to a second electrode 34b having nine features 35 according to an embodiment of the present disclosure. Figure 3C This is a top view showing the residual droplet 26b moving from the second electrode to the third electrode 34c according to an embodiment of the present disclosure, thereby leaving residual microdroplets (microdroplets) 26a within or on the feature. To prevent the microdroplets from evaporating in air, they can be surrounded by other immiscible liquids (such as silicone oil). It should be understood that the number of features on the electrodes can be any integer. Figures 3A to 3CAs shown, nine features are used in the second electrode. However, it should be understood that this number is arbitrarily chosen to describe the example embodiment and should not be limiting. It should also be understood that each electrode (e.g., the first, second, and third electrodes) may have the same number of features, or they may have different numbers of features. Reference Figures 3A to 3C The feature is shown as having a square shape; however, it should be understood that the shape shown is not limiting, and any other shape is equally applicable, such as circles, rectangles, ovals, ellipses, polygons, and other shapes.

[0078] Note that the electrodes according to embodiments of this disclosure can be arranged in various configurations, and the electrodes can have many shapes. For example, the electrodes can have polygonal shapes (e.g., squares, rectangles, triangles, etc.), circular shapes, oval shapes, etc. The configuration can be a checkerboard configuration or other geometric configurations.

[0079] Patterned features can be implemented in different ways. In one embodiment, the surface of the dielectric layer is selectively divided into hydrophobic and hydrophilic regions. Within a certain range of the surface area ratio between the hydrophobic and hydrophilic regions, and within a certain range of the interfacial tension between the droplet and the environment (oil or air), the droplet can be removed from the hydrophobic regions, leaving residual droplets (microdroplets) in the hydrophilic regions. The volume of the residual droplets (microdroplets) is defined by the size of the hydrophilic feature and the contact angle of the droplet on the surface of the environment (air or oil). Figure 4A This is a cross-sectional view of a portion of the EWOD device 40A according to an embodiment of this disclosure. (Refer to...) Figure 4A The EWOD device 40A includes a substrate structure having a first substrate (not shown) and a second substrate (not shown), a first dielectric layer 43a disposed on the first substrate, and a second dielectric layer 47a disposed on the second substrate (not shown). A droplet 26 is disposed between the first dielectric layer 43a having a first surface 44 and the second dielectric layer 47a having a second surface facing the first surface. In one embodiment, the first surface of the first dielectric layer includes a plurality of hydrophilic regions 48a protruding above the surface of the first dielectric layer 43a. The surface of the first dielectric layer 43a is coated with a hydrophobic film, i.e., the protruding hydrophilic regions 48a are surrounded by interstitial hydrophobic surface regions. The protruding hydrophilic regions 48a can be patterned such that residual droplets (microdroplets) of the droplet disposed on each hydrophilic region have a desired volume. Each protruding hydrophilic region can have a polygonal shape (e.g., square, rectangular), oval, circular, elliptical, and other shapes. In this disclosure, the term "protruding hydrophilic region" may also be referred to as a "spot" or "island". In some embodiments, the second dielectric layer 47a is made of glass coated with a hydrophobic film. In some embodiments, the first and second dielectric layers are formed separately, and such as in... Figure 2A , 2C Spacers, such as those shown and described in Figures 2D and 2E, are formed on a first dielectric layer or a second dielectric layer. The first and second dielectric layers are then bonded together to form... Figure 4A The structure shown has a space between the first and second dielectric layers that serves as a channel for droplets 26. In some embodiments, the second dielectric layer 47a is coated with a conductive layer of a metallic material serving as a common electrode (e.g., a ground electrode) and a hydrophobic film on the conductive layer. In some embodiments, the second dielectric layer may include, for example, Figure 2D and 2E The multiple sensors shown and described. Protruding hydrophilic regions can be formed on the surface of the first dielectric layer 43a using conventional semiconductor manufacturing techniques. Some of these semiconductor manufacturing techniques will be described in detail in the Examples section below. EWOD 40A also includes an array of electrodes (not shown) embedded within the first dielectric layer, the second dielectric layer, or both the first and second dielectric layers, which generate a moving electric field in response to a time-varying voltage provided by control circuitry. The droplet 26 moves across the entire surface of the first dielectric layer by the moving electric field, leaving residual small portions (microdroplets) 26' of the droplet 26 on the protruding hydrophilic region 48a.

[0080] Figure 4B This is a cross-sectional view of a portion of an EWOD device 40B according to another embodiment of this disclosure. (Refer to...) Figure 4B EWOD device 40B includes and Figure 4A The substrate structure shown is similar to that of a substrate structure, except that the first dielectric layer 43b includes a plurality of grooves (micropores) 49 of a certain depth instead of protruding areas. In one embodiment, the second surface of the second dielectric layer is hydrophobic, and the first dielectric layer includes a plurality of grooves (micropores) 49 of a certain depth. The terms “groove,” “recess,” and “micropore” are used interchangeably herein. Each groove also has an opening defined by its length and width. In some embodiments, the length and width are less than 1 micrometer. By carefully selecting the shape of the micropores and / or changing the inner surfaces (sidewalls and / or bottom) of the micropores from hydrophobic to hydrophilic through chemical (i.e., by surface treatment) or electrical (i.e., by electrowetting) means, a portion of the water-containing droplets spontaneously infiltrates into the micropores and tends to remain in the micropores even after the electric field is removed. Within a certain range, based on the area ratio between the flat hydrophobic surface of the first dielectric layer and the micropore, and the interfacial tension between the droplet and the environment (oil or air), the droplet can be removed from the micropore, leaving (depositing) residual small droplets (microdroplets) within the micropore. The volume of the residual small droplets (microdroplets) is determined by the predetermined micropore size and the contact angle of the droplet at the micropore opening. Combined with the above... Figure 4ASimilar in structure, the second dielectric layer 47b may be coated with a conductive layer of metallic material serving as a common electrode (e.g., a ground electrode) and a hydrophobic film on the conductive layer. Recessed hydrophilic regions can be formed in the first dielectric layer 43b using conventional semiconductor manufacturing techniques. The EWOD 40B also includes an array of electrodes (not shown) embedded within or in both the first and second dielectric layers, which generate a moving electric field in response to a time-varying voltage provided by control circuitry (not shown). The droplet 26 moves across the entire surface of the first dielectric layer by the moving electric field, leaving small residual portions (microdroplets) 26' of the droplet 26 on the surface of the micropores. In some embodiments, the micropore array can be obtained using photolithographic patterning and etching processes. Some of these semiconductor manufacturing techniques will be described in detail in the Examples section below.

[0081] Figure 4C yes Figure 4A A perspective view of a portion of the EWOD device 40A shown. (Reference) Figure 4C A group of circular cylindrical spots (protruding hydrophilic regions) 48a are arranged in an array. As used herein, the terms "spot," "island," and "protruding region" are used interchangeably. The spots 48a are spaced apart and isolated from each other by interstitial hydrophobic surfaces 44. Note that although the spots 48a are shown as having a circular cylindrical shape in the example shown, those skilled in the art will understand that the spots 48a may also have other shapes, such as rectangular, square, or oval cylindrical shapes.

[0082] Figure 4D yes Figure 4B A perspective view of a portion of the EWOD device 40B shown. (Reference) Figure 4D A group of circular cylindrical grooves (micropores) 49 are arranged in an array. The micropores 49 are filled with a hydrophobic material and spaced apart from each other by interstitial hydrophobic surfaces 46. In some embodiments, the hydrophobic material filling the micropores 49 has an upper surface flush with the upper surface of the interstitial hydrophobic surfaces 46. It should be noted that although the micropores 49 are shown as circular cylindrical pores, depending on the application, the micropores 49 may also be rectangular, square, or oval cylindrical pores.

[0083] Figure 4E This is a perspective view of a portion of an EWOD device according to an embodiment of this disclosure. (Refer to...) Figure 4E The group of hydrophilic surfaces 491 are surrounded (side-connected) by the hydrophobic gap region 461 on the surface of the dielectric layer 43, that is, the hydrophilic surface 491 is flush with the surface of the hydrophobic gap region 461 on the surface of the dielectric layer 43.

[0084] According to this disclosure, a large number of microdroplets of uniform size can be used for droplet digital PCR on a microfluidic chip. Due to the small volume of each sample and the DNA concentration below a certain level required to satisfy a Poisson distribution, each sample (microdroplet) will have one DNA molecule or no DNA molecule. Individual DNA molecules within a target region can be amplified on each sample in an environment (e.g., oil) by thermally cycling the samples (microdroplets) using conventional PCR or incubating them at a specific temperature using isothermal PCR. After reading the DNA concentration of the final droplet by optical detection or pH measurement using an integrated on-chip ion-sensitive field-effect transistor (ISFET) sensor, the absolute amount of target DNA in the sample (microdroplet) array can be quantified, and then the DNA concentration in the host droplet can be calculated using absolute DNA quantification. The terms “sample,” “residual small droplet,” “small portion of droplet,” and “microdroplet” are used interchangeably herein and refer to a small droplet formed from a host droplet according to an embodiment of this disclosure.

[0085] According to this disclosure, droplets containing multiple different DNA targets can be dispensed onto a region of a single microfluidic chip, and then the droplets can be moved to a next region by electrowetting, where a large number of samples (copies of the DNA targets) are generated from the droplets for sample detection or measurement. In some embodiments, the next region to which the droplets are moved may include multiple hydrophilic regions spaced apart by interstitial hydrophobic surfaces. The multiple hydrophilic regions may be combined Figure 4A and 4B Hydrophilic regions are shown and described. Each hydrophilic region may include an ion-sensitive field-effect transistor (ISFET) sensor configured to measure the pH of a sample disposed thereon. That is, a single microfluidic chip may include an array of ISFET sensors, each ISFET sensor associated with one of the samples in the droplet. This arrangement of a single microfluidic chip enables the formation of multiple samples (microdroplets) from the droplet by electrowetting and the measurement of the samples by an array of ISFET devices integrated on the chip. According to this disclosure, the ISFET sensor array embedded in the hydrophilic regions facilitates the simultaneous measurement of different targets from the droplet with high sensitivity and accuracy.

[0086] Figure 5 This is a cross-sectional view of an ISFET device 50 according to an embodiment of this disclosure. Reference Figure 5The ISFET device 50 is a metal-oxide-semiconductor (MOS) transistor, which can be a p-channel MOS field-effect transistor (MOSFET) or an n-channel MOSFET transistor fabricated using standard CMOS manufacturing processes. In the following description, according to an exemplary embodiment of this disclosure, an n-channel MOS transistor (NMOS) is used. However, it should be noted that the choice of NMOS or PMOS depends solely on the chosen process or substrate and is not limiting. Reference Figure 5 The ISFET device 50 has a substrate 52, a source region S and a drain region D formed in the substrate, a dielectric layer 53 on the substrate, and a floating gate G formed within or on the dielectric layer 53. The ISFET device also includes a sensing membrane 55 on the floating gate G and below a droplet 56, and a reference electrode 57 that is wholly or partially immersed in the droplet 56 and spaced apart from the sensing membrane 55. The sensing membrane can include any material that provides sensitivity to hydrogen ion concentration (pH), such as silicon nitride, silicon oxynitride, etc. Other sensing membranes sensitive to other ions can be used, as is known to those skilled in the art.

[0087] Still referencing Figure 5 The ISFET device 50 also includes a voltage source Vgs configured to provide a voltage Vgs between the sample and the source region S, and a voltage source Vds configured to provide a voltage Vds between the source and drain regions. When the voltage Vgs is greater than the threshold voltage Vth of the ISFET device, current will conduct through the channel between the source and drain regions. The amount of current Ids flowing between the source and drain regions represents the concentration or pH value of the sample 56. In one embodiment, the source region and the substrate have the same potential, such as ground potential.

[0088] exist Figure 5 In the example shown, one ISFET device is used to measure the ion concentration of droplet 56. However, it should be understood that this number is chosen only to describe exemplary embodiments and should not be limiting. In some embodiments, more than one ISFET device may be used to measure the ion concentration of droplet 56. In other words, each spot or microwell may have multiple ISFET devices.

[0089] In some implementation schemes, in the corresponding appendix Figure 4C and 4DThe diagram illustrates that each spot in a spot array or each micro-hole in a micro-hole array can have more than one ISFET device to improve measurement sensitivity and accuracy. The number of ISFET devices available per spot or micro-hole depends on conventional CMOS fabrication processes and application requirements. The power supply voltage provided to the ISFET devices via electrical connections can be implemented using conventional CMOS fabrication processes and is not described here for simplicity. In some embodiments, the ISFET devices can be integrated as sensors onto the second substrates 28d and 28e, as shown below. Figure 2D and 2E As shown.

[0090] Figure 6A This is a simplified top view of an integrated on-chip laboratory device 60A according to an embodiment of this disclosure. Reference Figure 6A The integrated on-chip laboratory device 60A includes a substrate structure having: a droplet receiving region 61 configured to receive one or more droplets 26; a reagent receiving region 62 configured to receive one or more reagents 63; a mixing region 64 configured to mix the droplets 26 with one or more reagents 63 to obtain mixed droplets 263; and an EWOD device array configured to split the droplets (mixed or unmixed) into multiple microdroplets and amplify the microdroplets. In one embodiment, the EWOD device array may have a first heating element configured to heat the microdroplets to a first temperature for amplifying the microdroplets and a second heating element for annealing the amplified microdroplets. In one embodiment, the on-chip laboratory device 60 may also include a sensor array, each sensor associated with a sample and configured to measure the concentration or pH of the microdroplets. In one embodiment, the droplet receiving region 61 may have Figure 1A and 1B The device structure is shown. In one embodiment, the reagent receiving area 62 may have... Figure 1A and 1B The device structure is shown. In other words, the integrated on-chip laboratory device 60 can be operable to move one or more droplets and one or more reagents to a mixing region 64, and control the mixing of droplets and reagents according to a user-provided software program. In one embodiment, the EWOD device array may include multiple EWOD devices arranged in a regular pattern, each EWOD device may have... Figures 2A to 2C The devices shown have similar or identical structures. In some implementations, each EWOD device may include multiple ISFETs. Examples of ISFET devices are shown in... Figure 5 As shown in the diagram. The integrated on-chip laboratory device 60A may also include a waste (collection) area 66 for collecting residual portions of droplets after they have formed in the EWOD device array and / or for collecting droplets after they have been processed and measured. Figure 6AIn the example shown, two electrodes are used in the droplet receiving region 61, eight electrodes are used in the upper part of the reagent receiving region 62, eight electrodes are used in the lower part of the reagent receiving region 62, eight electrodes are used in the mixing region 64, and an array of four EWOD devices is used. However, it should be understood that these numbers are arbitrarily chosen for describing exemplary embodiments and should not be limiting.

[0091] In some embodiments, the integrated on-chip laboratory device 60A may further include control circuitry 67 configured to provide control signals to droplet receiving region 61, reagent receiving region 62, mixing region 64, EWOD device array, and waste region 66 for moving droplets 61, reagents 63, mixed droplets 263, fragmented droplets (i.e., microdroplets), and residual portions of droplets after passing through the EWOD device array. In one embodiment, the integrated on-chip laboratory device 60A may include input / output (IO) ports 68 configured to interface with a host computer. In one embodiment, the host computer may be a separate or external processor configured to provide control signals to the integrated on-chip laboratory device 60. In another embodiment, the host computer may be integrated into the same package as the integrated on-chip laboratory device 60. Many variations, modifications, and alternatives will be recognized by those skilled in the art. Still refer to Figure 6A The control circuit 67 can be located remotely from the integrated on-chip laboratory device 60A and can communicate with the integrated on-chip laboratory device 60A via an input-output port or a serial interface port. In one embodiment, the integrated on-chip laboratory device 60A may include a sample inlet for receiving liquid samples and transferring the samples to a droplet receiving region 61. The integrated on-chip laboratory device 60A may include a reagent inlet for receiving one or more reagents and one or more reservoirs in the reagent receiving region 62 for storing the received reagents. In one embodiment, the integrated on-chip laboratory device 60A may also include a first heating block "heater 1" formed below the surface of the mixing region 64 within the substrate structure for maintaining and / or changing the incubation temperature of the mixed droplets 263. In one embodiment, the integrated on-chip laboratory device 60A may also include a second heating block "heater 2" formed below the surface of the EWOD device array within the substrate structure for maintaining and / or changing the incubation temperature of the microdroplets. The first heating block and the second heating block are formed of metal or polysilicon wires, metal or polysilicon layers, and polysilicon layers that can convert electrical energy from signals received from control circuit 67 into heat energy. In this disclosure, all operations of receiving droplets, mixing droplets with reagents, heating droplets, splitting droplets into multiple microdroplets, testing microdroplets, and discarding microdroplets after testing are collectively referred to as manipulating droplets.

[0092] Figure 6B This is a simplified top view of an example arrangement of electrodes 60B in an integrated on-chip laboratory device 60B according to an embodiment of this disclosure. Reference Figure 6B , Figure 6A The integrated on-chip laboratory device electrode 60B includes a first conductive strip array 611 having strips 611a, 611b, 611c, 611d, 611e, 611f, and 611g arranged parallel to each other, for example, along a first direction from a first region (droplet receiving region) toward a third region (EWOD device array). The first strip array 611 is controlled by control circuitry 67 and configured to generate a moving electric field to transport droplets along the first direction. Electrode 60B also includes a second parallel strip array 612 having strips 612a, 612b, 612c, and 612d arranged parallel to each other, for example, along a second direction perpendicular to the first direction. The second array 612 is configured to generate a moving electric field to transport droplets along the second direction toward a second region (mixing region). The first array intersects with and is spaced apart from the second array by an insulating layer; that is, the first array and the second array are arranged in different layers separated by at least one insulating layer. Figure 6B In the example shown, the second array 612 is arranged in the reagent receiving region for delivering reagents toward the mixing region. However, it should be understood that the second array 612 may also be arranged in the droplet receiving region, in the mixing region, and in the EWOD device array to generate a moving electric field and power to move droplets, mixed droplets, and microdroplets along the second direction. In some other embodiments, electrode 60B may include [missing information - likely related to bonding / contact]. Figure 1A and 1B The electrode array shown and described is similar to the electrode array shown. Those skilled in the art will recognize many variations, modifications, and alternatives.

[0093] Figure 7A This is a simplified cross-sectional view showing a multilayer ceramic substrate 70A according to an embodiment of the present disclosure. (Reference) Figure 7AThe multilayer ceramic substrate 70A is a laminated structure comprising multiple layers 71, 72, 73, 74, 75, and 76. The multilayer ceramic substrate 70A has multiple micropads 711 disposed on a first surface of a first layer (top layer) 71 and multiple contact pads 761 disposed on a second surface of a second layer (bottom layer) 76. The first and second surfaces are opposite to each other. The multilayer ceramic substrate 70A also includes multiple vias 716 passing through the first layer 71, the inner layers 72 to 75, and the second layer 76, the vias 716 being configured to connect the contact pads to one or more micropads. The multilayer ceramic substrate 70A may further include interconnect layers disposed between the inner layers. The interconnect layers may be configured to connect some of the vias together. For example, interconnect layer 734 connects vias 716a, 716b, and 716c together. In one embodiment, the micropads 711 may be configured as an array of driving electrodes and reference electrodes for moving or manipulating droplets. In one embodiment, the control pad 711 may be electrically coupled to a control board that provides electrical signals to the micropad to manipulate droplets disposed on the first layer. In one embodiment, the multilayer ceramic substrate 70A may include a glass-ceramic material.

[0094] In some embodiments, layers 71 to 76 may have the same thickness. In other embodiments, layers 71 to 76 may have different thicknesses. The layers, including interconnect layers and vias, can be fabricated simultaneously (at the same time) using the same manufacturing process, or they can be fabricated sequentially using the same or different processes. These layers are then assembled (laminated) together under pressure and high temperature to form a multilayer ceramic substrate 70A.

[0095] In one embodiment, the multilayer ceramic substrate 70A is configured as an EWOD substrate having micropads 711 on a top layer 71 serving as driving electrodes and contact pads on a bottom layer 76, the contact pads being connected to a control board to provide electrical control signals to the micropads.

[0096] In one embodiment, the micropad 711 may protrude from the upper surface of the first layer (top layer) 71. In another embodiment, the micropad 711 may be integrated into the first layer 71 and have a substantially flush (coplanar) upper surface with the upper surface of the first layer 71.

[0097] In one embodiment, the top layer 71 may be coated with an inorganic dielectric layer, such as a stack of alternating layers of silicon dioxide (SiO2), silicon nitride (SiN), or SiO2 and SiN. In another embodiment, the top layer 71 may be coated with an organic layer, such as polyimide. Silicon dioxide and / or silicon nitride can be deposited on the top layer 71 by silicon vapor deposition, atomic layer deposition, or flowable film deposition processes. The organic layer or polyimide layer can be formed by deposition, coating, or adhesive bonding processes. A hydrophobic layer is then formed on the inorganic or organic layer.

[0098] Figure 7B This is a simplified cross-sectional view showing a multilayer ceramic substrate 70B configured as an insert substrate for a single-layer substrate 71B according to an embodiment of the present disclosure. (Refer to...) Figure 7B The multilayer ceramic substrate 70B can be compared with the above references. Figure 7A The multilayer ceramic substrate 70A shown and described is the same. Figure 7A and 7B The difference lies in the fact that the multilayer ceramic substrate 70B is not directly used as an EWOD device for manipulating droplets. Instead, the multilayer ceramic substrate 70B is used as an insertion substrate for the monolayer substrate 71B.

[0099] The monolayer substrate 71B is designed for single-use (i.e., low-cost) applications and has a first surface 771, a second surface 772 opposite to the first surface, and a plurality of contact holes 773 extending from the first surface to the second surface and aligned with micropads 711 of the multilayer ceramic substrate 70B. The monolayer substrate 71B also includes an electrode array 781 disposed on the first surface 771 and a plurality of conductive features 774 filling the contact holes 773. Each conductive feature includes a protrusion 775 projecting from the second surface 772. In one embodiment, the upper surfaces 776 of the protrusions are substantially coplanar to allow for good electrical contact with the micropads 711. In one embodiment, the first surface 771 of the monolayer substrate 71B may be coated with an inorganic dielectric layer (e.g., silicon dioxide (SiO2), silicon nitride (SiN), or a stack of alternating layers of SiO2 and SiN). In one embodiment, the first surface 771 may be coated with an organic layer, such as polyimide. Silicon dioxide and / or silicon nitride can be deposited on the first surface 771 of the monolayer substrate 71B using silicon vapor deposition, atomic layer deposition, or flowable film deposition processes. An organic layer or polyimide layer can be formed by deposition, coating, or adhesive bonding processes. A hydrophobic layer is then formed on the inorganic or organic layer. Because the multilayer ceramic substrate 70B is used as an insertion substrate, it can be fixed or permanently mounted on test equipment.

[0100] In one embodiment, the disposable single-layer substrate 71B may be a single-layer substrate comprising a ceramic material. Figure 7C This is a perspective view of an exemplary single-layer ceramic substrate 70C according to an embodiment of the present disclosure. Reference Figure 7C The single-layer substrate 70C includes an insulating layer (e.g., a ceramic layer) 781C sandwiched between a first copper layer 782C and a second copper layer 783C.

[0101] In one embodiment, the disposable monolayer substrate 71B may be a monolayer substrate comprising silicon and / or glass materials. In another embodiment, the disposable monolayer substrate 71B may be a monolayer substrate comprising organic or plastic materials. When the monolayer substrate comprises silicon or glass, or a combination of silicon and glass, the manufacturing cost may be higher than when the monolayer substrate is made of ceramic, but it offers advantages for droplets, such as a smoother surface (lower surface roughness Ra) and better gap tolerances. In other words, a monolayer substrate made of silicon or glass, or both, has superior performance compared to monolayer substrates made of ceramic, organic, or plastic materials. In some embodiments, the monolayer substrate is made of a rigid material suitable for providing a uniform load distribution on the micropads 711.

[0102] Figure 7D This is a simplified cross-sectional view showing a multilayer ceramic substrate 70D connected to a single-layer substrate 71D via a fastening member 79D according to an embodiment of the present disclosure. Reference Figure 7D A multilayer ceramic substrate 70D, serving as an insert between the instrument and a single-layer substrate 71D, and a single-layer substrate 71D, serving as an EWOD substrate for manipulating droplets, are mechanically connected by a mechanical fastening member 79D. The multilayer ceramic substrate 70D may be similar to the insert substrate 70B described above, and the single-layer substrate 71D may be similar to the single-layer substrate 71B described above. The mechanical fastening member 79D may include a set of clamps, a set of screws and bolts, or other fasteners known in the art.

[0103] Figure 8A This is a simplified cross-sectional view showing a socket 80A for supporting an integrated circuit in accordance with an embodiment of the present disclosure. In one embodiment, the socket 80A can similarly be used as a connector. Figure 7B The multilayer ceramic substrate 70B shown is an insertion substrate between the instrument and the disposable substrate. (Reference) Figure 8A The socket 80A may include a socket body 80, which has a plurality of upper contact pins 81, a plurality of lower contact pins 83, and a plurality of through holes 85 connecting the upper contact pins 81 and the lower contact pins 83. The upper contact pins 81 correspond to... Figure 7B The micropad 711 shown has lower contact pins 83 configured to interface with pins of high-density electronic device packages such as ball grid arrays or pad grid arrays. The socket 80A is constructed to accommodate a high-density package containing integrated circuits (e.g., control circuitry that provides electrical signals to a disposable substrate to manipulate droplets). In other words, Figure 7B The conductive feature 774 of the disposable substrate 71B is aligned with the upper contact pin 81. In one embodiment, each upper contact pin 81 may include a ball pin 82. In one embodiment, the socket 80A can be soldered to a printed circuit board (e.g., a control board) to serve as an insert (interconnect) between the instrument and the disposable substrate.

[0104] Figure 8B An exemplary ball grid array socket according to an embodiment of the present disclosure is shown, which can be used as an insert (interconnect) between test instruments and a disposable substrate.

[0105] Figure 8C An exemplary pad grid array socket according to an embodiment of the present disclosure is shown, which can be used as an insert (interconnect) between test instruments and a disposable substrate.

[0106] Figure 8D A simplified cross-sectional view of a monolayer substrate 81D including multiple sensors according to an embodiment of the present disclosure is shown. The monolayer substrate 81D includes a substrate 810, an electrode array 881 disposed on an upper surface of the substrate, each electrode in the electrode array including a conductive feature 874 extending through the substrate and a protrusion 875 projecting from a lower surface. The protrusions have upper surfaces that are substantially coplanar with each other. The monolayer substrate 81D also includes a recess 820 configured to receive a porous dielectric layer 830 having a first surface 831 flush with the upper surface of the electrode array 881 and a second surface opposite to the first surface 831. The monolayer substrate 81D also includes multiple sensors 840 in contact with the second surface and multiple actuation electrodes 850 configured to generate an electric field to transport a portion of a droplet through the porous dielectric layer to the sensors for testing or measurement. The sensors may be references. Figure 5 The described MOS sensor or FET sensor. Electrode array 881 and actuation electrode 850 are... Figure 7B The micro pad 711 of the insertion substrate 70B or Figure 8A The chip packaging carrier 80A has electrical and physical contacts at its contact pins 81 or contact balls 82. In one embodiment, the electrode array and actuation electrodes receive electrical signals via a device packaged in a BGA or LGA supported by the carrier 80A.

[0107] Figure 9 This is a simplified flowchart illustrating a method 90 for manufacturing an apparatus for manipulating droplets according to an embodiment of the present disclosure. (See reference) Figure 9Method 90 may include, at 901, providing a multilayer ceramic substrate having a first surface and a second surface opposite to the first surface, the first surface including an array of electrodes, and the second surface having a plurality of contact pads electrically contacting one or more of the driving electrodes. At 903, the method may include forming a dielectric layer on the first surface of the multilayer ceramic substrate. In one embodiment, the dielectric layer may include an inorganic material, such as silicon dioxide and / or silicon nitride. In one embodiment, the dielectric layer may include an organic material, such as polyimide. In one embodiment, forming the dielectric layer may include a spin-coating or adhesive bonding process. At 905, the method may further include forming a hydrophobic layer on the dielectric layer. In some embodiments, the multilayer ceramic substrate thus formed can be used directly to manipulate droplets. In other embodiments, the method may further include, at step 907, providing a second substrate having a third surface including a plurality of electrodes, a fourth surface opposite to the third surface, a plurality of through-holes extending from the third surface to the fourth surface, and a plurality of conductive features filling the through-holes. Each conductive feature has a protrusion above the fourth surface and aligned with one of the plurality of driving electrodes. In this configuration, a second substrate is operable to receive droplets in place of a multilayer ceramic substrate, which serves as an insert between the test instrument and the second substrate. In some embodiments, the second substrate is a disposable substrate. To maintain low cost, the second substrate is a single-layer substrate. In one embodiment, the second substrate is a single-layer ceramic substrate. In another embodiment, the second substrate includes a dielectric layer and a hydrophobic layer on the dielectric layer. The method may further include, in step 909, attaching the second substrate to the multilayer ceramic substrate using fastening members such as clamps, screws and bolts, springs, etc.

[0108] Figure 10 This is a simplified flowchart illustrating a method 100 for manufacturing an apparatus for manipulating droplets according to an embodiment of the present disclosure. (See reference) Figure 10 Method 100 may include, at 1001, providing a receptacle for receiving a package containing an integrated circuit. The receptacle may include a receptacle body having a plurality of upper contact pins disposed on a flat upper surface of the receptacle body and a plurality of lower contact pins disposed on a flat lower surface of the receptacle body. The lower surface of the receptacle body may be soldered onto a control board. At 1003, the method may further include providing a substrate having a first surface including a plurality of electrodes, a second surface opposite the first surface, a plurality of through-holes extending through the substrate, and a plurality of conductive features filling the through-holes, each of the conductive features including an upper portion projecting from the second surface and an upper surface substantially coplanar with each other. The conductive features are substantially aligned with the upper contact pins. At 1005, the method may further include attaching the substrate to the receptacle using a fastening member, such that the conductive features make electrical and physical contact with the upper contact pins of the receptacle.

[0109] Figure 11 This is a controllable reference based on the embodiments of this disclosure. Figure 1A A simplified schematic diagram of the mobile computing device 110 of the apparatus or device described in -B, 2A-2I, 3A-C, 4A-B, 5, 6A-B, 7A-D, 8A-C, 9, and 10. (Refer to...) Figure 11 The mobile computing device 110 may include a monitor 1110, a computing electronic device 1120, a user output device 1130, a user input device 1140, a communication interface 1150, etc.

[0110] The computing electronic device 1120 may include one or more processors 1160 that communicate with a plurality of peripheral devices via a bus subsystem 1190. These peripheral devices may include user output devices 1130, user input devices 1140, communication interfaces 1150, and storage subsystems such as random access memory (RAM) 1170 and disk drives 1180.

[0111] User input device 1140 may include any type of device and interface for inputting information into computer device 1120, such as keyboard, keypad, touch screen, mouse, trackball, trackpad, joystick and other types of input devices.

[0112] User output device 1130 may include any type of device for outputting information from computing electronics 1120, such as a display (e.g., monitor 1110).

[0113] Communication interface 1150 provides an interface to other communication networks and devices. Communication interface 1150 can be used as an interface for receiving data from and sending data to other systems. For example, communication interface 1150 may include a USB interface for communicating with devices used to manipulate droplets.

[0114] RAM 1170 and disk drive 1180 are examples of tangible media configured to store data, such as embodiments of this disclosure, including executable computer code, human-readable code, etc. Other types of tangible media include floppy disks, removable hard disks, optical storage media (e.g., CD-ROMs, DVDs, and barcodes), semiconductor memories (e.g., flash memory), non-transitory read-only memory (ROM), battery-supported volatile memory, network storage devices, etc. RAM 1170 and disk drive 980 can be configured to store basic programming and data constructs that provide the functionality of this invention.

[0115] The software code modules and instructions that provide the functionality of this disclosure can be stored in RAM 1170 and disk drive 1180. These software modules can be executed by processor 1160.

[0116] Still referencing Figure 11 The device for manipulating droplets may include an interface port configured to provide communication with a mobile computing device 110. In some embodiments, the mobile computing device 110 may provide command and control signals via the device's interface port to control the signal levels of electrodes in the device. In some embodiments, the device for manipulating droplets may include, for example... Figure 2A-2I The substrate structure described in 3A-3C and 4A-4E and Figure 5 One or more of the ISFET devices. The device is configured to receive droplets and provide a pH value associated with the droplets.

[0117] Example

[0118] The following embodiments are provided as examples, but do not limit the claimed invention.

[0119] Example 1

[0120] This embodiment discloses the formation of electrodes within a dielectric layer. In one embodiment, a first dielectric layer is formed on a substrate. A patterned photoresist layer is then formed on the dielectric layer. A metal layer or a doped polysilicon layer is then deposited on the first dielectric layer using the patterned photoresist layer as a mask. The patterned photoresist layer is then removed. Subsequently, a second dielectric layer is formed on the metal layer to form an electrode as described above. Figures 2A to 2C The structure shown.

[0121] In another embodiment, a first dielectric layer is formed on a substrate. A metal layer is then deposited on the first dielectric layer. Subsequently, a patterned photoresist layer is formed on the metal layer. An etching process is then performed on the metal layer using the patterned photoresist layer as a mask. The patterned photoresist layer is then removed. Thereafter, a second dielectric layer is formed on the metal layer to form a shape as shown in the image. Figures 2A to 2C The structure shown.

[0122] Example 2

[0123] This embodiment discloses a novel apparatus and method for generating large arrays of nanoliters (10⁻⁶) from droplets via electrowetting. -9 liters or nL) and pills (10 -12 Extremely small droplets (microdroplets) within a predetermined volume range (in liters or pL). This novel device can be used with... Figure 4A or Figure 4B The EWOD device shown is similar to or the same as the device. (Refer to...) Figures 3A to 3CThis illustrates a delivery mechanism for dispensed droplets. The droplets move on the surface of a dielectric layer having an array of raised hydrophilic regions (spots) or on the surface of a dielectric layer having an array of micropores, each micropore having a hydrophilic bottom and sidewalls. The raised (protruding) hydrophilic regions (spots) and / or micropores are spaced apart from each other by interstitial hydrophobic surfaces. In one embodiment, the raised hydrophilic regions and / or micropores are circular or oval. In another embodiment, the raised hydrophilic regions and / or micropores are polygonal (rectangular, square, hexagonal) in shape. In one embodiment, the height of the raised hydrophilic regions (spots) and / or micropores is between 1 nanometer and 100 micrometers, preferably between 1 micrometer and 10 micrometers, more preferably below 10 nanometers. In one embodiment, the raised hydrophilic regions and / or micropores are square in shape, with a width or length between 1 nanometer and 100 micrometers, preferably between 1 micrometer and 10 micrometers, more preferably below 10 nanometers. In one embodiment, the raised hydrophilic regions and / or micropores have a circular shape and a diameter between 1 nanometer and 100 micrometers, preferably between 1 micrometer and 10 micrometers, and more preferably below 10 nanometers.

[0124] Example 3

[0125] This embodiment discloses a novel apparatus and method for mixing droplets with one or more reagents and delivering the mixed droplets to an EWOD array to obtain multiple microdroplets. In one embodiment, the microdroplets produced by the novel apparatus and method have a uniform size. The novel apparatus may be an integrated laboratory-on-a-chip device similar to or the same as shown in FIG. 6. In some embodiments, the novel apparatus may include different regions, such as a droplet discharge region for receiving droplets, a reagent discharge region for receiving reagents, a mixing region for mixing droplets with reagents, and an array of IWOD devices for generating multiple samples of uniform size. In one embodiment, the apparatus may also include a temperature regulating element for performing conventional PCR or incubating the microdroplets at a predetermined temperature (isothermal PCR). In one embodiment, each IWOD region may have an array of raised (protruding) hydrophilic surface regions or spots (e.g., Figure 4A and 4C (as shown) or arrays of recessed hydrophilic surface regions or micropores (such as... Figure 4B and Figure 4D (As shown). In some embodiments, each spot or microwell includes one or more ISFETs for measuring the ion concentration or pH of the associated microdroplet. In one embodiment, the device may include an interface port configured to communicate with an external host that collects the measured pH of the sample and calculates the DNA concentration of the droplets.

[0126] Example 4

[0127] This embodiment discloses the formation of a multilayer ceramic substrate having a dielectric layer formed thereon and a hydrophobic layer formed on the dielectric layer. Figure 12 This is a cross-sectional view illustrating the process of manufacturing a multilayer ceramic substrate structure 1200. The multilayer ceramic substrate structure 1200 includes a first ceramic layer 12a having a plurality of first through-holes 121, a second ceramic layer 12b including a plurality of second through-holes 123 and a first patterned metal layer 124, and a third ceramic layer 12c including a plurality of third through-holes 125 and a second patterned layer 126. A method of forming the multilayer ceramic substrate may include sequentially laminating the first ceramic layer 12a and the second ceramic layer 12b, and then the method may further include laminating the third layer 12c with the laminated layers 12a and 12b. The method may further include filling the through-holes with a conductive material such as a metal (Ag, Pd, Cu, Ni, etc.). The method may continue to laminate more ceramic layers and fill the through-holes until a predetermined thickness and a usable substrate are obtained. Then, a bottom layer 12bb including contact pads and through-holes filled with conductive material and a top layer 12tt including micro-pads and through-holes filled with conductive material are subsequently attached to opposite sides of the substrate to obtain a multilayer ceramic substrate, for example... Figure 7A or Figure 7B Multilayer ceramic substrates 70A or 70B.

[0128] Subsequently, a dielectric layer can be deposited on the formed multilayer ceramic substrate using a conventional deposition process (e.g., chemical vapor deposition). The dielectric layer may include silicon oxide, silicon nitride, fluorinated silicate glass (FSG), or organosilicon glass (OSG). The dielectric layer is then coated with a hydrophobic material. A spin-coating process can be used to coat the hydrophobic material, in which the hydrophobic material is sprayed onto the surface of the dielectric layer. For example, the surface of the dielectric layer is exposed to a hydrophobic solution at a specific temperature for a specific time period. Alternatively, the hydrophobic material can be formed on the dielectric layer using a deposition process (e.g., chemical vapor deposition). The hydrophobic material may include organic or inorganic materials, such as octadecyltrichlorosilane, perfluorodecyltrichlorosilane, other fluorinated layers, such as tetrafluoroethylene, etc.

[0129] Example 5

[0130] This embodiment discloses the formation of a single-layer substrate having an electrode array disposed on its upper surface, a plurality of contact holes, and a plurality of conductive components filling the contact holes, such as... Figure 7B The substrate 71B shown is shown. Figures 13A to 13H A cross-sectional view illustrating an intermediate step in the process of manufacturing a single-layer substrate is shown. In one embodiment, a substrate 130 is provided. The substrate 130 may have an insulating layer 131 (silicon oxide, silicon nitride, or other insulating material) disposed or grown thereon, such as... Figure 13A As shown. (Refer to...) Figure 13B A patterned photoresist layer 132, including multiple openings 133, is formed on a substrate 130. Deep reactive ion etching (RIE) is performed on the substrate 130 using the photoresist layer 132 as a mask to form multiple trenches 134 in the substrate. The patterned photoresist layer and insulating layer (if present) are then removed. Figure 13C As shown. In Figure 13D In this process, a metal layer (tungsten, aluminum, copper, etc.) is deposited on a substrate to form an upper metal layer 136, and trenches are filled to form multiple features 137. A planarization (chemical mechanical polishing) process is performed on the upper metal layer 136 to provide a smooth surface. Figure 13E In this process, a planarization process (e.g., chemical mechanical polishing) is performed on the bottom surface of the substrate, such that the bottom surfaces of the conductive features are exposed and flush with each other (in a substantially coplanar relationship). Figure 13F In this process, plasma etching, wet etching, or a combination of both are performed to remove a portion of the bottom of the substrate 130 that exposes the conductive features 137. Figure 13G In this process, a patterned photoresist layer 138 is deposited on an upper metal layer 136, and a plasma etching or wet etching process is performed on the upper metal surface 136 using the patterned photoresist layer 138 as a mask to separate the upper metal layer 136 into a plurality of electrodes 139 spaced apart from each other. Figure 13H A cross-sectional view of the monolayer substrate formed after the above steps have been performed is shown. The monolayer substrate can be... Figure 7B The disposable substrate 71B shown may be made of ceramic, resin, glass, plastic or other insulating materials.

[0131] Although the process described herein is described with respect to a specific number of steps performed in a particular order, it is contemplated that additional steps not explicitly shown and / or described may be included. Furthermore, it is contemplated that fewer steps than shown and described may be included without departing from the scope of the described embodiment (i.e., one or some of the described steps may be optional). Additionally, it is contemplated that the steps described herein may be performed in a different order than that described.

[0132] It should be understood that the embodiments and implementations described herein are for illustrative purposes only, and various modifications or changes made thereto will be suggested by those skilled in the art and will be included within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

[0133] It should be understood that the above description is intended to illustrate rather than limit. Many embodiments will be apparent to those skilled in the art upon reading the above description. Therefore, the scope of the invention should not be determined by reference to the above description, but rather by reference to the full scope of the appended claims and their equivalents.

[0134] While the foregoing disclosure illustrates illustrative aspects of this disclosure, it should be noted that various changes and modifications may be made herein without departing from the scope of this disclosure as defined by the appended claims. The functions, steps, and / or actions of the method claims according to aspects of this disclosure described herein do not need to be performed in any particular order. Furthermore, although elements of this disclosure may be described or claimed in the singular, plural forms are conceivable unless a limitation on the singular is expressly stated.

[0135] For all the flowcharts in this article, it should be understood that many steps can be combined, executed in parallel, or executed in different orders without affecting the functionality implemented.

Claims

1. An apparatus for manipulating droplets, the apparatus comprising: A socket for supporting an integrated circuit package includes a plurality of contact pads disposed on a flat surface of the socket and configured to receive electrical signals from the integrated circuit. and A disposable monolayer substrate having a first surface, a second surface, a plurality of electrodes on the first surface, and a plurality of through-holes extending through the monolayer substrate, the monolayer substrate being configured to receive droplets, wherein the disposable monolayer substrate does not comprise a plurality of substrate layers, and wherein each electrode includes a conductive feature through a through-hole in the substrate and a protrusion protruding from the second surface of the disposable monolayer substrate, wherein the conductive feature is made of the same conductive material as the electrode; and Mechanical fastening components are used to connect the single-layer substrate to the socket. The plurality of electrodes on the first surface of the monolayer substrate are electrically coupled to the plurality of contact pads in the socket through the vias and are configured to manipulate the droplets disposed on the first surface of the monolayer substrate by means of the electrical signals provided by the integrated circuit.

2. The apparatus according to claim 1, wherein, The socket is a ball grid array socket.

3. The apparatus according to claim 1, wherein, The socket is a pad grid array socket.

4. The apparatus according to claim 1, wherein, The conductive material contains copper.

5. The apparatus according to claim 1, wherein, The single-layer substrate includes multiple sensors configured to detect the properties of the droplets.

6. The apparatus of claim 1, further comprising a counter substrate facing and spaced apart from the monolayer substrate, the counter substrate and the monolayer substrate forming a channel for the droplet.

7. The apparatus according to claim 6, wherein, The opposing substrate includes a plurality of sensors configured to detect the properties of the droplets.

8. The apparatus of claim 5, wherein the disposable single-layer substrate includes a groove configured to accommodate a porous dielectric layer in which the plurality of sensors are located.

9. The apparatus of claim 8, further comprising a plurality of actuating electrodes configured to transfer a portion of a droplet through the porous dielectric layer to the plurality of sensors.

10. A method of manufacturing an apparatus for manipulating droplets, the method comprising: A receptacle is provided for accommodating a package containing an integrated circuit, the receptacle including a receptacle body, the receptacle body including a plurality of upper contact pins disposed on a flat surface of the receptacle body and configured to receive electrical signals from the integrated circuit; A disposable substrate is provided, the substrate having a first surface including a plurality of electrodes, a second surface opposite to the first surface, a plurality of through holes extending through the disposable substrate, and a plurality of conductive features, each of the conductive features having a protrusion protruding from the second surface electrode, wherein the disposable substrate is configured to receive droplets on the first surface, wherein the disposable substrate is a disposable single-layer substrate that does not contain multiple substrate layers, and the conductive features and the electrodes are made of the same conductive material; as well as The disposable substrate is attached to the surface of the socket body using a fastening member, such that the conductive features of the disposable substrate are in electrical and physical contact with the upper contact pins of the socket, so as to provide the electrical signal to the electrode for manipulating the droplet.

11. The method according to claim 10, wherein, The socket is a ball grid array socket.

12. The method according to claim 10, wherein, The socket is a pad grid array socket.

13. The method according to claim 10, wherein, The conductive material contains copper.

14. The method of claim 10, wherein, The conductive feature is aligned with the upper contact pin.

15. The method of claim 10, wherein the disposable single-layer substrate includes a groove configured to accommodate a porous dielectric layer in which a plurality of sensors are located.

16. The method of claim 15, further comprising a plurality of actuating electrodes configured to transfer a portion of a droplet through the porous dielectric layer to the plurality of sensors.

17. An apparatus for manipulating droplets, comprising: Disposable single-layer substrate, the disposable single-layer substrate comprising: A first surface, a second surface, and a plurality of through holes extending from the first surface to the second surface; The electrode array on the first surface of the disposable single-layer substrate; and Multiple conductive features coupled to the electrode array and filling the vias, each of the multiple conductive features including a protrusion extending from a second surface of the monolayer substrate and aligned with one of the first electrode arrays, wherein the vias are aligned with the first electrode arrays; The disposable monolayer substrate is a substrate for receiving droplets on the first surface of the disposable monolayer substrate, and the electrode array is configured to receive electrical signals to electrically manipulate the droplets; The disposable single-layer substrate does not contain multiple substrate layers, and the conductive features and the electrodes use the same conductive material; and The disposable single-layer substrate is configured to be connected to another device via a fastening member to receive electrical signals.

18. The apparatus of claim 17, further comprising a opposing substrate having a first surface facing the disposable monolayer substrate and a second surface spaced apart from the disposable monolayer substrate to form an air gap for the droplet.

19. The apparatus according to claim 18, wherein, The opposing substrate includes a common electrode disposed on the second surface.

20. The apparatus according to claim 19, wherein, The opposing substrate also includes a plurality of sensors configured to detect the properties of the droplets.

21. The apparatus of claim 17, further comprising: A common electrode on a dielectric layer, wherein the dielectric layer is located on the disposable monolayer substrate; as well as The substrate has a second surface facing the first surface of the disposable monolayer substrate and spaced apart from the disposable monolayer substrate to form an air gap for the droplet.

22. The apparatus according to claim 21, wherein, The opposing substrate includes a plurality of sensors configured to detect the properties of the droplets.

23. The apparatus of claim 17, wherein the disposable single-layer substrate includes a groove configured to accommodate a porous dielectric layer in which a plurality of sensors are located.

24. The apparatus of claim 23, further comprising a plurality of actuating electrodes configured to transfer a portion of a droplet through the porous dielectric layer to the plurality of sensors.

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