Micro-droplet control chip, micro-droplet control device, preparation method and application of micro-droplet control chip

Abstract

The invention provides a micro-droplet control chip, a micro-droplet control device, a preparation method and application thereof. The micro-droplet control device comprises a control unit and a reaction unit, and the control unit is connected with the reaction unit; the reaction unit comprises a circuit board and a micro-droplet control chip, and the circuit board is bonded with the micro-droplet control chip; the micro-droplet control chip comprises a silicon-based substrate chip and an ITO (Indium Tin Oxide) chip; the silicon-based substrate chip comprises a silicon-based substrate, an electrode layer, a dielectric layer and a first hydrophobic layer; the ITO chip comprises an ITO substrate and a second hydrophobic layer; and the silicon-based substrate chip and the ITO chip are bonded to form an internal flow channel. Compared with the prior art, the device has at least one of the following beneficial effects: the device is small in size, simple to operate, small in reagent amount required by reaction, simple in manufacturing process, low in manufacturing cost, capable of simultaneously controlling multiple groups of liquid drops, and capable of realizing high flux, accurate temperature measurement and control and real-time feedback adjustment.

Description

Technical Field

[0001] The present application relates to the field of microfluidic control, and relates to a micro-droplet manipulation chip, a device, a preparation method and uses thereof, and specifically relates to a micro-droplet manipulation device and uses thereof, as well as a micro-droplet manipulation chip and a preparation method thereof. Background Art

[0002] As microreactors, droplets have the advantages of high throughput, small size, and rapid reaction. They are widely used in biosynthesis, biochemical analysis, drug encapsulation, drug transport and release. Therefore, precise control of microdroplets is particularly important.

[0003] At present, droplet control is mainly in the form of microfluidic chips, with piezoelectric, electrowetting, dielectrophoresis, ultrasound, air pump and other driving methods. Among them, electrowetting microfluidic chips are widely used in the field of biosynthesis. For example, in DNA synthesis, compared with the traditional well plate method, the use of electrowetting chips has the advantages of high degree of automation, small size, small amount of reagents required, and low cost. However, existing electrowetting chips generally have the disadvantages of single function, complex process and low flux. For example, the electrowetting electrode can only realize the movement of droplets but not temperature control; secondly, the electrode structure design is single and the number is small, which cannot meet the needs of high-throughput droplet microreactions.

[0004] Therefore, the current micro-droplet manipulation device and micro-droplet manipulation chip still need to be improved. Summary of the invention

[0005] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a micro-droplet manipulation device and its use, as well as a micro-droplet manipulation chip and its preparation method, so as to at least alleviate or even solve at least one of the problems raised in the above background technology to a certain extent.

[0006] In the first aspect of the present invention, the present invention provides a micro-droplet manipulation device, which includes a control unit and a reaction unit, wherein the control unit is connected to the reaction unit; the reaction unit includes a circuit board and a micro-droplet manipulation chip, and the circuit board is bonded to the micro-droplet manipulation chip; the micro-droplet manipulation chip includes a silicon-based substrate chip and an ITO chip; the silicon-based substrate chip includes a silicon-based substrate, an electrode layer, a dielectric layer, and a first hydrophobic layer; the ITO chip includes an ITO substrate and a second hydrophobic layer; the silicon-based substrate chip is bonded to the ITO chip to form an internal flow channel.

[0007] Furthermore, the micro-droplet manipulation chip is bonded to the circuit board in a manner that includes at least one of wire bonding and through silicon via bonding.

[0008] Furthermore, the micro-droplet manipulation device further includes a liquid medium and reaction droplets, and the liquid medium and reaction droplets are located in the internal flow channel of the micro-droplet manipulation chip; the control unit is electrically connected to the circuit board to control the movement, fusion and separation of the reaction droplets.

[0009] Furthermore, in the micro-droplet manipulation chip, the ITO chip includes a liquid inlet, a liquid outlet and a liquid medium inlet; wherein the number of the liquid inlet, the liquid outlet and the liquid medium inlet is independently one or more.

[0010] Furthermore, in the micro-droplet manipulation chip, the electrode layer includes a plurality of independent electrodes, and the plurality of independent electrodes are configured to independently control voltage.

[0011] Furthermore, the material forming the independent electrode includes at least one of Au, Pt and Cu.

[0012] Furthermore, in the micro-droplet manipulation chip, the independent electrode includes at least one of an electrowetting electrode, a heating electrode, and a temperature sensing electrode.

[0013] Furthermore, in the micro-droplet manipulation chip, the graphic structure of the electrode layer includes at least one of a rectangular shape, a forked finger shape, a winding shape, a spiral shape, and other shapes.

[0014] Furthermore, the electrowetting electrode and the heating electrode are the same electrode.

[0015] In another aspect of the present invention, the present invention provides a micro-droplet manipulation chip, and the micro-droplet manipulation chip is as described above.

[0016] In another aspect of the present invention, the present invention provides a method for preparing the above-mentioned micro-droplet manipulation chip, the method comprising: preparing a silicon-based substrate chip; preparing an ITO chip; bonding the silicon-based substrate chip and the ITO chip together, and utilizing the bonds and interfaces between the silicon-based substrate chip and the ITO chip to form an internal flow channel that is closed on all sides.

[0017] Furthermore, preparing the ITO chip includes coating a second hydrophobic layer material on an ITO substrate, and providing a liquid inlet, a liquid outlet, and a liquid medium inlet.

[0018] Furthermore, preparing the silicon-based substrate chip includes: sequentially forming a silicon oxide layer, an electrode layer, a dielectric layer and a first hydrophobic layer on the silicon-based substrate.

[0019] According to another aspect of the present invention, the present invention provides a method for controlling microdroplets using the above-mentioned microdroplet manipulation device, the method comprising injecting a liquid medium into the internal flow channel of the microdroplet manipulation chip; injecting reaction droplets into the internal flow channel of the microdroplet manipulation chip; controlling the electrodes in the microdroplet manipulation chip by a control unit to control the movement, fusion, and separation of the reaction droplets to form product droplets; wherein the liquid medium and the reaction droplets are immiscible with each other.

[0020] Furthermore, the method further includes providing a driving signal through the control unit to control the product droplets to move to the liquid outlet and extract the product droplets.

[0021] Furthermore, the liquid medium includes at least one of oil or glycerol.

[0022] Furthermore, the driving signal includes at least one of a pulse signal and a direct current signal.

[0023] Furthermore, the reaction droplets contain at least one of DNA molecules, PCR products, PCR reaction reagents, and magnetic beads.

[0024] The present invention has at least one of the following beneficial effects:

[0025] 1. The micro-droplet manipulation device of the present invention performs micro-droplet manipulation on a micro-droplet manipulation chip, has a small size and is easy to operate; requires a small amount of reagents for biosynthesis reactions, and has low cost;

[0026] 2. The micro-droplet manipulation device of the present invention integrates a heating electrode and a temperature sensing electrode on the micro-droplet manipulation chip, which can provide the required temperature for the micro-droplet reaction and provide real-time feedback adjustment, simplifying the operation and reducing the use of equipment;

[0027] 3. The electrowetting electrode and the heating electrode on the micro-droplet manipulation chip of the present invention can be the same electrode, which simplifies the process steps and saves chip manufacturing costs;

[0028] 4. The micro-droplet manipulation chip of the present invention can be used to manufacture multiple groups of array electrodes. Through silicon via packaging technology, multiple groups of droplets can be manipulated simultaneously, which has the advantage of high throughput.

[0029] 5. The electrodes of the micro-droplet manipulation chip of the present invention adopt structures such as forked fingers and windings, so that the electrowetting electrodes, heating electrodes, and temperature sensing electrodes are arranged on the same layer structure, which can accurately measure and control temperature changes and has the advantages of simple process, space saving, and low cost.

[0030] The above summary is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments and features described above, further aspects, embodiments and features of the present application will be readily apparent by reference to the accompanying drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In the accompanying drawings, unless otherwise specified, the same reference numerals throughout the multiple drawings represent the same or similar parts or elements. These drawings are not necessarily drawn to scale. It should be understood that these drawings only depict some embodiments disclosed in the present application and should not be regarded as limiting the scope of the present application.

[0032] Figure 1 A schematic diagram of the structure of a micro-droplet manipulation device according to an embodiment of the present invention;

[0033] Figure 2 This is a schematic diagram of the structure of a micro-droplet manipulation chip according to an embodiment of the present invention;

[0034] Figure 3 A schematic diagram of the structure of a micro-droplet manipulation device according to an embodiment of the present invention;

[0035] Figure 4 This is a schematic diagram of the structure of a micro-droplet manipulation chip according to an embodiment of the present invention;

[0036] Figure 5 A schematic diagram of an interdigitated electrode structure according to an embodiment of the present invention;

[0037] Figure 6 A schematic diagram of a wire-wound electrode structure according to an embodiment of the present invention;

[0038] Figure 7 is a schematic diagram of an interdigitated electrode structure array according to an embodiment of the present invention;

[0039] Figure 8 A flow chart of a method for preparing a micro-droplet manipulation chip according to an embodiment of the present invention;

[0040] Fig. 9 This is a process flow chart for preparing a micro-droplet manipulation chip in Example 1 of the present invention;

[0041] Fig.10 This is a process flow chart for preparing a micro-droplet manipulation chip in Example 2 of the present invention.

[0042] Explanation of the accompanying drawings: 1000-micro-droplet manipulation chip, 2000-circuit board, 3000-control unit, 3100-power supply, 3200-control component, 100-silicon-based substrate chip, 110-silicon-based substrate, 120-silicon oxide layer, 130-electrode layer, 140-dielectric layer, 150-first hydrophobic layer, 200-ITO chip, 210-ITO substrate, 220-second hydrophobic layer, 230-liquid inlet, 240-liquid outlet, 250-liquid medium inlet, 10-internal flow channel, 20-bonding glue, 30-metal copper. DETAILED DESCRIPTION

[0043] In order to more clearly understand the technical features, purposes and beneficial effects of the present invention, the technical scheme of the present invention is now further described in detail. Hereinafter, only some exemplary embodiments are simply described. As those skilled in the art will appreciate, the described embodiments may be modified in various ways without departing from the spirit or scope of the present application. Therefore, the drawings and descriptions are considered to be exemplary and non-restrictive in nature.

[0044] In one aspect of the present invention, the present invention provides a micro-droplet manipulation device, referring to Figure 1 The device includes a control unit 3000 and a reaction unit connected to each other, the reaction unit includes a circuit board 2000 and a micro-droplet manipulation chip 1000, and the circuit board 2000 in the reaction unit is bonded to the micro-droplet manipulation chip 1000. The micro-droplet manipulation chip 1000 includes reaction droplets and a liquid medium, and the control unit 3000 is electrically connected to the circuit board 2000 of the reaction unit to control the movement, fusion or separation of the reaction droplets on the micro-droplet manipulation chip 1000. The control unit 3000 includes a power supply 3100 and a control component 3200, and the control component 3200 includes an electronic device, such as a computer, a mobile phone, etc.; refer to Figure 2 The micro-droplet manipulation chip 1000 includes a silicon-based substrate chip 100 and an ITO chip 200. The silicon-based substrate chip 100 includes a silicon-based substrate 110, a silicon oxide layer 120, an electrode layer 130, a dielectric layer 140, and a first hydrophobic layer 150. The ITO chip 200 includes an ITO substrate 210 and a second hydrophobic layer 220, and the silicon-based substrate chip 100 and the ITO chip 200 are bonded to form an internal flow channel 10. The micro-droplet manipulation device has at least one of the following beneficial effects: small size, simple operation, small amount of reagents required for reaction, simple manufacturing process, low manufacturing cost, can manipulate multiple groups of droplets at the same time, can achieve high throughput, accurately measure and control temperature and real-time feedback adjustment.

[0045] For ease of understanding, the following first briefly describes the principle by which the micro-droplet manipulation device can achieve the above beneficial effects:

[0046] The micro-droplet manipulation device of the present invention adopts a micro-droplet manipulation chip to control the reaction droplets, and the micro-droplet manipulation chip of the present invention adopts electrode layer structures such as interdigitated fingers and windings, and arranges the electrowetting electrode and electrodes with multiple functions such as heating electrodes and temperature sensing electrodes on the same layer structure, wherein the electrowetting electrode and the heating electrode can be reused to realize the multifunctionality of the electrode, save space, simplify the process steps, and reduce the manufacturing cost; and the electrode layer integrated on the micro-droplet manipulation chip of the present invention can have multiple functions, for example, it can provide the required temperature for the micro-droplet reaction and provide real-time feedback regulation; in addition, multiple groups of array electrodes can be made on the micro-droplet manipulation chip of the present invention, and multiple groups of droplets can be manipulated simultaneously to achieve high throughput.

[0047] According to an embodiment of the present invention, the connection method between the reaction unit and the control unit 3000 in the micro-droplet manipulation device of the present invention can be as follows: Figure 3 As shown, the control component 3100 in the control unit 3000 is a computer. In addition, the bonding method of the micro-droplet manipulation chip 1000 and the circuit board 2000 is not particularly limited. For example, in a specific embodiment, the bonding method includes but is not limited to wire bonding, through silicon via bonding or a combination thereof.

[0048] According to an embodiment of the present invention, the micro-droplet manipulation device includes a liquid medium and reaction droplets for reaction, wherein the liquid medium and the reaction droplets are both located in the internal flow channel 10 of the micro-droplet manipulation chip 1000; the control unit 3000 is electrically connected to the circuit board 2000 to control the movement, fusion and separation of the reaction droplets in the micro-droplet manipulation chip 1000.

[0049] According to an embodiment of the present invention, in the micro-droplet manipulation chip 1000, the ITO chip 200 has a liquid inlet 230, a liquid outlet 240 and a liquid medium inlet 250. Among them, the liquid inlet 230 is used to inject reaction droplets, and the number of liquid inlets 230 is not particularly limited. Specifically, it can be set according to the type of reaction droplets, for example, it can be one or more; its position is not particularly limited, and it can be located around or in the middle of the ITO chip 200. The liquid outlet 240 is used to output and collect reaction products, and the number of liquid outlets 240 is not particularly limited, and it can be one or more; its position is not particularly limited, and it can be located around or in the middle of the ITO chip 200, but in a specific embodiment, it is preferred that multiple liquid outlets 240 are set on the same side to facilitate the output and collection of product droplets. The liquid medium inlet 250 is used to inject liquid medium, and the number of liquid medium inlets 250 is not particularly limited, and it can be one or more, preferably one; its position is not particularly limited, and it can be located around or in the middle of the ITO chip 200. In a specific embodiment, refer to Figure 4, shows a schematic structural diagram of a micro-droplet manipulation chip, which includes two liquid inlets 230 , two liquid outlets 240 and a liquid medium inlet 250 .

[0050] According to an embodiment of the present invention, the electrode layer 130 of the chip of the present invention may include a plurality of independent electrodes, each of which includes two interfaces; wherein the independent electrode is made of an electrode material, and the type of the electrode material is not particularly limited, for example, including but not limited to Au, Pt, Cu or a combination thereof. In addition, the type and function of the independent electrode are also not particularly limited, for example, including but not limited to an electrowetting electrode, a heating electrode, a temperature sensing electrode or a combination thereof. In a specific embodiment, the electrode layer 130 of the chip of the present invention may simultaneously include at least one of an electrowetting electrode, a heating electrode, and a temperature sensing electrode, and the electrowetting electrode and the heating electrode may be the same independent electrode. For example, in a specific embodiment, the ITO chip may be grounded by utilizing the conductive function of the ITO substrate in the ITO chip, and only one interface of the independent electrode may be energized to realize the electrowetting function; or by energizing the two interfaces of the independent electrode, applying current to the independent electrode, and using the resistance of the independent electrode to generate Joule heat to realize the temperature heating function; in addition, since temperature changes will cause changes in the resistance of the metal, and the resistance-temperature is linearly related, by measuring the resistance at both ends of the independent electrode, the temperature can be calculated and then the temperature can be monitored to realize the temperature sensing function.

[0051] According to the embodiment of the present invention, the graphic structure of the electrode in the electrode layer 130 is not particularly limited. For example, the graphic structure may include but is not limited to a rectangular shape, a forked shape, a winding shape, a spiral shape, and other shapes, or a combination thereof. For example, it may include but is not limited to Figure 5 The interdigitated electrode structure shown, and Figure 6 In addition, the arrangement of the electrode layer 130 is not particularly limited, and may include, for example, an array, a single electrode, or other arrangements. Figure 7 A schematic diagram of an array of interdigitated electrode structures is shown. And Figure 5 , Figure 6 and Figure 7 Each independent electrode shown in can be independently selected from an electrowetting electrode, a heating electrode or a temperature sensing electrode. In addition, the size of the chip of the present invention is not particularly limited and can be determined according to the reaction type and the amount of the desired reaction droplet product.

[0052] In another aspect of the present invention, the present invention provides a micro-droplet manipulation chip, which is the same as the micro-droplet manipulation chip included in the above-mentioned micro-droplet manipulation device, specifically, as Figure 2As shown, the micro-droplet manipulation chip 1000 is composed of a silicon-based substrate chip 100 and an ITO chip 200. The silicon-based substrate chip 100 is composed of a silicon-based substrate 110, a silicon oxide layer 120, an electrode layer 130, a dielectric layer 140, and a first hydrophobic layer 150 connected in sequence, and the ITO chip 200 is composed of an ITO base 210 and a second hydrophobic layer 220, and the silicon-based substrate chip 100 and the ITO chip 200 are bonded together by bonding glue, and an internal cavity enclosed on all sides is formed at the bonding position, which is the internal flow channel 10.

[0053] In another aspect of the present invention, the present invention provides a method for preparing the above-mentioned micro-droplet manipulation chip. Figure 8 The method includes separately preparing a silicon-based substrate chip 100 and an ITO chip 200, and bonding the silicon-based substrate chip 100 and the ITO chip 200 together using a bonding glue 20, wherein the order of preparing the silicon-based substrate chip 100 and the ITO chip 200 is not subject to any special restrictions. In addition, the bond and interface between the silicon-based substrate chip 100 and the ITO chip 200 form a cavity enclosed on all sides, namely, an internal flow channel 10, and the size of the internal flow channel 10 is not subject to any special restrictions, and can be specifically determined according to the size of the chip. The method has at least one of the following beneficial effects: simple process steps, low manufacturing cost, space saving, small size of the prepared chip, and the ability to manipulate multiple groups of droplets at the same time.

[0054] According to an embodiment of the present invention, the step of preparing the ITO chip 200 includes coating the second hydrophobic layer 220 on the ITO substrate 210, and arranging thereon one or more liquid inlets 230, liquid outlets 240, and liquid medium inlets 250. The second hydrophobic layer 220 is not particularly limited as long as it has a hydrophobic effect. For example, in a specific embodiment, the material of the second hydrophobic layer 220 may include but is not limited to Teflon.

[0055] According to an embodiment of the present invention, the step of preparing a silicon-based substrate chip 100 includes sequentially forming a silicon oxide layer 120, an electrode layer 130, a dielectric layer 140, and a first hydrophobic layer 150 on a silicon-based substrate 110. Specifically, in the process of forming the silicon oxide layer, the silicon oxide layer 120 can be deposited on the silicon-based substrate 110 to form the silicon oxide layer 120 on the silicon-based substrate 110, wherein the deposition method includes but is not limited to chemical vapor deposition; or, in the process of forming the silicon oxide layer 120, the silicon oxide layer 120 can also be formed by performing a through silicon via process on the silicon-based substrate 110, firstly etching the silicon-based substrate 110 into a through hole structure, then growing a silicon oxide insulating layer on the surface of the silicon-based substrate 110 and the sidewall of the through hole through thermal oxidation treatment, then filling the through hole through an electroplating process, and finally chemical mechanical planarization to obtain the silicon-based substrate 110 that has completed the through silicon via process. In the process of forming the electrode layer, the electrode layer 130 can be formed on the surface of the silicon oxide layer 120 by ion beam etching, stripping process or a combination thereof. The independent electrode types, graphic structures, electrode materials, etc. of the electrode layer 130 have been described in detail above and will not be repeated here. In the process of forming the dielectric layer, the dielectric layer 140 material can be deposited on the electrode layer 130. The deposition method includes but is not limited to chemical vapor deposition, and the dielectric layer 140 material is not particularly limited, for example, but not limited to silicon oxide. In the process of forming the first hydrophobic layer, the material of the first hydrophobic layer 150 can be coated on the surface of the dielectric layer 140 to form the first hydrophobic layer 150, and a silicon-based substrate chip 100 is obtained, wherein the material of the first hydrophobic layer 150 is not particularly limited, for example, including but not limited to Teflon.

[0056] According to another aspect of the present invention, the present invention provides a method for controlling microdroplets using the above-mentioned microdroplet manipulation device, the method comprising injecting a liquid medium from a liquid medium inlet 250 into the internal flow channel 10 of a microdroplet manipulation chip 1000; then injecting a reaction droplet from a liquid inlet 230 into the internal flow channel 10 of the microdroplet manipulation chip; and then providing a driving signal to a circuit board 2000 connected to its electrical signal through a control unit 3000, and transmitting the signal to the microdroplet manipulation chip 1000 to control the movement, fusion, and separation of the reaction droplets in the microdroplet manipulation chip 1000, and finally forming product droplets. Among them, the liquid medium and the reaction droplet are insoluble in each other, and the injection method of the reaction droplet is not particularly limited, for example, it can be injected using a pipette, a syringe, a rubber-tipped dropper, etc., or the electrode layer 130 can be powered to use the hydrophilicity of the electrode to suck the droplet into the internal flow channel 10. In addition, the method further includes providing a driving signal to the circuit board 2000 through the control unit 3000, controlling the product droplets to move to the liquid outlet 240, and extracting the product droplets at the liquid outlet 240, wherein the product droplets can be large droplets formed by fusion or small droplets formed by separation.

[0057] According to an embodiment of the present invention, the number of times the driving signal is provided by the control unit 3000 is not particularly limited, and can be one or more times. In addition, the intensity and duration of the driving signal are not particularly limited, and can be determined according to the type of reaction to be generated. In addition, when multiple driving signals are provided, the intensity and duration of each driving signal can be the same or different. In a specific embodiment, the driving signal includes but is not limited to a pulse signal, a DC signal, or a combination thereof.

[0058] According to an embodiment of the present invention, the injection method of the reaction droplets is not particularly limited, and the reaction droplets can be directly injected into the internal flow channel 10 using a pipette, a syringe, a rubber-tipped dropper, etc., or the reaction droplets can be provided at the liquid inlet through the control unit 3000 to provide a driving signal, power up specific electrodes on the silicon-based substrate chip 100, change the wettability of the silicon-based substrate 110, and make the reaction droplets be sucked into the internal flow channel 10 by the hydrophilicity of the electrode for reaction.

[0059] According to an embodiment of the present invention, the type of liquid medium is not particularly limited, and can be specifically determined according to the reaction to be carried out, and can be incompatible with the reaction droplets. For example, in a specific embodiment, the liquid medium includes inorganic or organic substances, such as but not limited to oil, glycerol or a combination thereof. In addition, the reaction droplets are also not particularly limited, including but not limited to polymer materials. Specifically, the reaction droplets can contain DNA molecules, PCR products, PCR reaction reagents, magnetic beads or a combination thereof. It should be particularly noted here that in this application, the term "magnetic beads" should be understood in a broad sense: magnetic beads refer to "beads" that can be used for biological / chemical analysis and have multiple functions, such as magnetic beads that can be used for nucleic acid extraction, cell separation, protein separation and molecular detection, and the magnetic beads can be magnetic beads or non-magnetic beads.

[0060] Example

[0061] The method proposed in the present invention is described in detail below through specific examples. The methods used in the following examples are conventional methods unless otherwise specified, and the reagents used are commercially available reagents unless otherwise specified.

[0062] Example 1

[0063] This embodiment uses the method of the present invention to prepare a micro-droplet manipulation chip, and uses a micro-droplet manipulation device including the obtained micro-droplet manipulation chip to control micro-droplets.

[0064] 1. Preparation of microdroplet manipulation chip

[0065] ①Preparing a silicon-based substrate chip 100:

[0066] like Fig. 9 As shown in A, first, the silicon-based substrate 110 is etched into a through-hole structure, and then a silicon oxide insulating layer is grown on the surface of the silicon-based substrate 110 and the sidewall of the through-hole through thermal oxidation treatment, and then the through-hole is filled with metal copper 30 through electroplating, and finally chemical mechanical planarization is performed to obtain a silicon-based substrate that has completed the silicon through-hole process; an electrode layer 130 is formed on the surface of the silicon oxide layer 120 through ion beam etching or stripping process; a silicon oxide dielectric layer 140 is deposited on the electrode layer 130 through chemical vapor deposition; and a first hydrophobic layer 150 is spin-coated on the surface of the dielectric layer 140 to form a silicon-based substrate chip 100. In this embodiment, the electrowetting electrode, the heating electrode, and the temperature sensing electrode are all interdigitated electrodes, and the electrowetting electrode and the heating electrode share an electrode structure.

[0067] ②Preparation of ITO chip 200:

[0068] like Fig. 9 As shown in B, the second hydrophobic layer 220 is spin-coated on the ITO chip 200 substrate, and two liquid inlets 230 (a first liquid inlet and a second liquid inlet), two liquid outlets 240 (a first liquid outlet and a second liquid outlet) and a liquid medium inlet 250 are arranged thereon.

[0069] ③ Bonding the silicon-based substrate chip 100 and the ITO chip 200:

[0070] like Fig. 9 As shown in C in FIG. 1 , the silicon-based substrate chip 100 and the ITO chip 200 are bonded together by bonding glue 20 , and the bond and the interface form an internal flow channel 10 that is closed on all sides, thereby forming a micro-droplet manipulation chip 1000 .

[0071] 2. Micro-droplet manipulation device

[0072] The micro-droplet manipulation chip obtained in this embodiment is bonded to the circuit board 2000 to form a reaction unit, and the reaction unit is connected to the control unit 3000 including the control component 3200 and the power supply 3100. The circuit board 2000 is connected to the control unit 3000 circuit, and the obtained micro-droplet manipulation chip is connected to the circuit board 2000 through a metal lead circuit to form a micro-droplet manipulation device.

[0073] 3. Using the micro-droplet manipulation device in this embodiment to control the micro-droplets

[0074] The liquid medium used in this embodiment is glycerol, and the reaction droplets include a salt solution (first reaction droplet) containing PCA products and a salt solution (second reaction droplet) containing PCR reaction reagents. The liquid medium is allowed to fill the internal flow channel 10 of the chip through the liquid medium inlet 250, the first reaction droplet is allowed to enter the internal flow channel 10 through the first liquid inlet 230, and the second reaction droplet is allowed to enter the internal flow channel 10 through the second liquid inlet 230. Then, the control component 3200 in the control unit 3000 is used to apply a pulse signal to the micro-droplet manipulation chip 1000 on the circuit board 2000 to energize the electrowetting electrode to realize the electrowetting function, and control the first reaction droplet and the second reaction droplet to move to a specific electrode for fusion. The control component 3200 in the control unit 3000 is used to apply a pulse signal to the micro-droplet manipulation chip 1000 on the circuit board 2000 again to energize the heating electrode to realize the heating function of the electrode where the reaction droplet is located. Then, a pulse signal is applied to the micro-droplet manipulation chip 1000 on the circuit board 2000 again to energize the temperature sensing electrode to realize the temperature sensing of the electrode where the reaction droplet is located, so as to adjust the temperature required for the droplet micro-reaction through the control unit 3000. After the reaction is completed, the control component 3200 in the control unit 3000 is used to apply a pulse signal to the micro-droplet manipulation chip 1000 on the circuit board 2000 to realize the electrowetting function, control the droplets to separate and move to the first liquid outlet 240 and the second liquid outlet 240 to extract the product droplets.

[0075] Example 2

[0076] In this embodiment, a micro-droplet manipulation chip 1000 is prepared using the method of the present invention, and a micro-droplet manipulation device including the obtained micro-droplet manipulation chip is used to control micro-droplets.

[0077] 1. Preparation of micro-droplet manipulation chip 1000

[0078] ①Preparing a silicon-based substrate chip 100:

[0079] like Fig.10 As shown in A, a silicon oxide layer 120 is deposited on the silicon-based substrate 110 by chemical vapor deposition to form a silicon oxide layer 120. Then, an electrode layer 130 is formed on the surface of the silicon oxide layer 120 by ion beam etching or stripping process; a silicon oxide dielectric layer 140 is deposited on the electrode layer 130 by chemical vapor deposition; and a first hydrophobic layer 150 is spin-coated on the surface of the dielectric layer 140 to form a silicon-based substrate chip 100. In this embodiment, the electrowetting electrode, the heating electrode, and the temperature sensing electrode are all array interdigitated electrodes, and the electrowetting electrode and the heating electrode share an electrode structure.

[0080] ②Preparation of ITO chip 200:

[0081] like Fig.10 As shown in B, the second hydrophobic layer 220 is spin-coated on the ITO chip 200 substrate, and two liquid inlets 230 (a first liquid inlet and a second liquid inlet), two liquid outlets 240 (a first liquid outlet and a second liquid outlet) and a liquid medium inlet 250 are arranged thereon.

[0082] ③ Bonding the silicon-based substrate chip 100 and the ITO chip 200:

[0083] like Fig.10 As shown in C in FIG. 1 , the silicon-based substrate chip 100 and the ITO chip 200 are bonded together by bonding glue 20 , and the bond and the interface form an internal flow channel 10 that is closed on all sides, thereby forming a micro-droplet manipulation chip 1000 .

[0084] 2. Micro-droplet manipulation device

[0085] The micro-droplet manipulation chip obtained in this embodiment is bonded to the circuit board 2000 to form a reaction unit, and the reaction unit is connected to the control unit 3000 including the control component 3200 and the power supply 3100. The circuit board 2000 is connected to the control unit 3000 circuit, and the obtained micro-droplet manipulation chip is connected to the through-silicon via circuit of the circuit board 2000 to form a micro-droplet manipulation device.

[0086] 3. Using the micro-droplet manipulation device in this embodiment to control the micro-droplets

[0087] The liquid medium used in this embodiment is glycerol, and the reaction droplets include a salt solution containing magnetic beads (first reaction droplets) and a salt solution containing Oligo molecules (second reaction droplets). The liquid medium is allowed to fill the internal flow channel 10 of the chip through the liquid medium inlet 250, the first reaction droplet is allowed to enter the internal flow channel 10 through the first liquid inlet 230, and the second reaction droplet is allowed to enter the internal flow channel 10 through the second liquid inlet 230. Then, the control component 3200 in the control unit 3000 is used to apply a pulse signal to the micro-droplet manipulation chip 1000 on the circuit board 2000 to energize the electrowetting electrode to realize the electrowetting function, and control the first reaction droplet and the second reaction droplet to move to a specific electrode for fusion. The control component 3200 in the control unit 3000 is used to apply a pulse signal to the micro-droplet manipulation chip 1000 on the circuit board 2000 again to energize the heating electrode to realize the heating function of the electrode where the reaction droplet is located. Then, a pulse signal is applied to the micro-droplet manipulation chip 1000 on the circuit board 2000 again to energize the temperature sensing electrode to realize the temperature sensing of the electrode where the reaction droplet is located, so as to adjust the temperature required for the droplet micro-reaction through the control unit 3000. After the reaction is completed, the control component 3200 in the control unit 3000 applies a pulse signal to the chip on the circuit board 2000 to realize the electrowetting function, control the droplet separation and move to the first liquid outlet 240 and the second liquid outlet 240 to extract the product droplet.

[0088] In general, the micro-droplet manipulation device proposed in the present invention adopts a micro-droplet manipulation chip 1000 with a specific structure. The micro-droplet manipulation chip 1000 adopts electrode layer structures such as forked fingers and windings, and arranges the electrowetting electrode, the heating electrode, and the temperature sensing electrode on the same layer structure, wherein the electrowetting electrode and the heating electrode can be the same electrode, which saves space, simplifies the process steps, and reduces manufacturing costs; and the micro-droplet manipulation chip 1000 of the present invention integrates the heating electrode and the temperature sensing electrode, which can provide the required temperature for the micro-droplet reaction and provide real-time feedback and regulation; in addition, the micro-droplet manipulation chip 1000 of the present invention can be made into multiple groups of array electrodes, and multiple groups of droplets can be manipulated at the same time to achieve high throughput.

[0089] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. For those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive.

[0090] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A micro-droplet manipulation device, characterized in that: The micro-droplet manipulation device comprises a control unit and a reaction unit, wherein the control unit is connected to the reaction unit; The reaction unit includes a circuit board and a micro-droplet manipulation chip, and the circuit board is bonded to the micro-droplet manipulation chip; The micro-droplet manipulation chip comprises a silicon-based substrate chip and an ITO chip; The silicon-based substrate chip comprises a silicon-based substrate, an electrode layer, a dielectric layer, and a first hydrophobic layer; The ITO chip comprises an ITO substrate and a second hydrophobic layer; The silicon-based substrate chip and the ITO chip are bonded to form an internal flow channel.

2. The micro-droplet manipulation device according to claim 1, characterized in that: The micro-droplet manipulation chip is bonded to the circuit board in a manner that includes at least one of wire bonding and through silicon via bonding.

3. The micro-droplet manipulation device according to claim 1, characterized in that: The micro-droplet manipulation device further comprises a liquid medium and a reaction droplet, wherein the liquid medium and the reaction droplet are located in an internal flow channel of the micro-droplet manipulation chip; The control unit is electrically connected to the circuit board to control the movement, fusion and separation of the reaction droplets.

4. The micro-droplet manipulation device according to claim 1, characterized in that: In the micro-droplet manipulation chip, the ITO chip includes a liquid inlet, a liquid outlet, and a liquid medium inlet; Wherein, the number of the liquid inlet, the liquid outlet and the liquid medium inlet is independently one or more.

5. The micro-droplet manipulation device according to claim 1, characterized in that: In the micro-droplet manipulation chip, the electrode layer includes a plurality of independent electrodes, and the plurality of independent electrodes are configured to independently control voltage; The material forming the independent electrode includes at least one of Au, Pt and Cu.

6. The micro-droplet manipulation device according to claim 1, characterized in that: The micro-droplet manipulation chip meets at least one of the following conditions: The independent electrode includes at least one of an electrowetting electrode, a heating electrode, and a temperature sensing electrode; The graphic structure of the electrode layer includes at least one of a rectangular shape, an interdigitated shape, a winding shape, a spiral shape, and other shapes; Wherein, the electrowetting electrode and the heating electrode are the same electrode.

7. A micro-droplet manipulation chip, characterized in that: The micro-droplet manipulation chip is as described in any one of claims 1-6.

8. A method for preparing the micro-droplet manipulation chip according to claim 7, characterized in that: The method comprises: preparing silicon-based substrate chips; Preparing ITO chips; The silicon-based substrate chip and the ITO chip are bonded together, and an internal flow channel closed on all sides is formed by utilizing the bond and interface between the silicon-based substrate chip and the ITO chip.

9. The method according to claim 8, characterized in that The preparation of the ITO chip includes coating a second hydrophobic layer on an ITO substrate, and setting a liquid inlet, a liquid outlet and a liquid medium inlet.

10. The method according to claim 8, characterized in that The preparation of the silicon-based substrate chip includes: sequentially forming a silicon oxide layer, an electrode layer, a dielectric layer and a first hydrophobic layer on the silicon-based substrate.

11. A method for controlling micro-droplets using the micro-droplet manipulation device according to any one of claims 1 to 6, characterized in that: The method comprises: Injecting a liquid medium into the internal flow channel of the micro-droplet manipulation chip; injecting the reaction droplets into the internal flow channel of the micro-droplet manipulation chip; The control unit controls the electrodes in the micro-droplet manipulation chip to control the movement, fusion, and separation of the reaction droplets to form product droplets; Wherein, the liquid medium and the reaction droplets are immiscible with each other.

12. The method according to claim 11, characterized in that The method further includes providing a driving signal through the control unit to control the product droplets to move to the liquid outlet and extract the product droplets.

13. The method according to claim 12, characterized in that The method satisfies at least one of the following conditions: The liquid medium includes at least one of oil or glycerol; The driving signal includes at least one of a pulse signal and a DC signal; The reaction droplets contain at least one of DNA molecules, PCR products, PCR reaction reagents, and magnetic beads.

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