Electro-sorting chip, preparation method of electro-sorting chip and electro-sorting method

Abstract

The invention discloses an electric separation chip, a preparation method of the electric separation chip and an electric separation method.The electric separation chip comprises a chip body and a circuit board, the chip body comprises a sample loading structure and a separation structure, the sample loading structure is provided with a sample loading groove, an oil channel and an air channel, the sample loading groove is formed above the separation structure, and a liquid drop inlet is formed in the upper surface of the sample loading structure; one end of the air channel extends to the side wall of the loading structure, the other end of the air channel is communicated with the loading groove, the lower end of the loading groove is communicated with the sorting structure, one end of the oil channel extends to the side wall of the loading structure, and the other end of the oil channel is communicated with the sorting structure; and the circuit board is arranged on the sample loading structure and supplies power to the sorting structure. The device is convenient for sample loading and sample replacement.

Description

Technical Field

[0001] This invention relates to the field of electrical separation technology, and in particular to electrical separation chips, methods for preparing electrical separation chips, and electrical separation methods. Background Technology

[0002] The core principle of droplet electrosort is based on real-time detection and sorting of droplets using biochemical fluorescent labeling. When a droplet flows through the detection area, a photomultiplier tube (PMT) collects the data and quickly identifies the target droplet using machine learning classification or thresholding algorithms. The sorting actuator applies a controllable Coulomb force or dielectric force to the target droplet using a high-frequency electric field, causing it to deviate from the default flow path and enter the designated collection channel. This technology offers advantages such as non-contact operation, low shear force (reducing cell damage), high throughput (thousands to tens of thousands of droplets per second), and programmable control. Its throughput is 2-3 orders of magnitude higher than traditional multi-well plate methods, and it is widely used in single-cell sequencing, rare cell sorting, droplet digital PCR (ddPCR), and high-throughput drug screening.

[0003] Sample loading methods for electro-sorting chips mainly include pneumatic drive and syringe pump drive. Pneumatic drive facilitates sample loading; however, conventional centrifuge tube-based pneumatic drive methods can lead to abnormal droplet states after prolonged injection, such as large-area fusion. Syringe pump drive can maintain droplet stability to some extent; however, this method is more complex to operate when changing samples. Summary of the Invention

[0004] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes an electrical separation chip, a method for preparing the electrical separation chip, and an electrical separation method.

[0005] The solution to the technical problem of this invention is: Firstly, an electrical sorting chip is proposed, comprising: The chip body includes a sample loading structure and a sorting structure. The sample loading structure is provided with a sample loading groove, an oil channel, and an air channel. The sample loading groove is located above the sorting structure. The upper surface of the sample loading structure is provided with a droplet inlet, which is connected to the upper end of the sample loading groove. One end of the air channel extends to the side wall of the sample loading structure, and the other end is connected to the sample loading groove. The lower end of the sample loading groove is connected to the sorting structure. One end of the oil channel extends to the side wall of the sample loading structure, and the other end is connected to the sorting structure. A circuit board is disposed on the sample-upper structure and electrically connected to the sorting structure.

[0006] This invention has at least the following beneficial effects: During sample loading, the droplet suspension is added to the sample loading tank from the droplet inlet, and driving air pressure is applied to the sample loading tank through the air channel, causing the droplet suspension to be pressed down into the sorting structure located below. Because a direct downward pressure method is used for droplet loading, excellent droplet condition can be ensured. Since both the oil channel and the air channel extend to the side wall of the sample loading structure, and the oil phase is external, when replacing the electro-sorting chip, the oil pipe can be directly connected through the opening of the oil channel in the side wall of the sample loading structure. The air pressure supply structure can also be connected through the opening of the air channel in the side wall of the sample loading structure, making operation convenient.

[0007] As a further improvement to the above technical solution, the sample loading structure is a 3D printed part made of transparent material; the sorting structure is a part made of polydimethylsiloxane material; the chip body also includes a bonding layer, and the sample loading structure and the sorting structure are connected through the bonding layer.

[0008] As a further improvement to the above technical solution, the bonding layer is double-sided adhesive, the sorting structure is bonded to one side of the double-sided adhesive, and the other side of the double-sided adhesive is bonded to the sample loading structure; or the bonding layer is a glass plate, the sorting structure is bonded to one side of the glass plate, and the other side of the glass plate is bonded to the sample loading structure, the glass plate is provided with at least two through holes, and the lower end of the sample loading groove and the oil passage are respectively connected to the sorting structure through the through holes.

[0009] As a further improvement to the above technical solution, the electrical sorting chip also includes a power supply structure, which is disposed on the sample structure and connected to the circuit board. The power supply structure is used to connect to an external power supply, and the power supply structure is one of a power supply socket, a magnetic spring pin connector, or a contact conductive component.

[0010] As a further improvement to the above technical solution, the electrical sorting chip also includes a pneumatic supply structure. The pneumatic supply structure is provided with a pneumatic supply plug. The end of the air passage extending to the side wall of the sample loading structure is a pneumatic supply port. The pneumatic supply plug is directly connected to the pneumatic supply port or is connected to the pneumatic supply port through a soft plug adapter.

[0011] As a further improvement to the above technical solution, the electrical sorting chip also includes a wire magnet. A magnet groove is provided below the sample loading structure, and the wire magnet is disposed in the magnet groove and connected to the sorting structure.

[0012] As a further improvement to the above technical solution, the sample loading structure is provided with a viewing window, which extends through the upper and lower surfaces of the sample loading structure and is located above the sorting structure.

[0013] As a further improvement to the above technical solution, the chip body also includes a packaging plate, which is connected to the sample structure and covers the upper end of the window. The packaging plate is a colored glass component or a filter.

[0014] Secondly, a method for fabricating an electrical sorting chip is proposed, applicable to the electrical sorting chip described in any of the above technical solutions, wherein the method for fabricating the electrical sorting chip includes the following steps: The sample structure was fabricated using 3D printing. The sorting structure is assembled below the sample loading structure; The circuit board is mounted on the sample loading structure and soldered to the sorting structure.

[0015] Thirdly, an electrical sorting method is proposed, which uses an electrical sorting chip as described in any of the above technical solutions, and the electrical sorting method includes the following steps: The droplet suspension is added to the sample loading tank from the droplet inlet; A driving air pressure is applied to the air passage to press the droplet suspension in the sample loading tank down into the sorting structure; When a droplet enters the sorting structure, power is supplied to the sorting structure via a circuit board.

[0016] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly explained below. Obviously, the described drawings are only a part of the embodiments of the present invention, and not all of them. Those skilled in the art can obtain other design schemes and drawings based on these drawings without creative effort.

[0018] Figure 1 This is a schematic diagram of the sample loading structure of the electrical sorting chip according to an embodiment of the present invention; Figure 2 This is a top view of the sample structure of the electrical sorting chip according to an embodiment of the present invention; Figure 3 This is a schematic diagram of the state of droplets after sample loading using the electro-sorting chip of this embodiment of the invention; Figure 4 This is a flowchart of the fabrication method of the electrically sorting chip according to an embodiment of the present invention; Figure 5 This is a flowchart of the electrical sorting method according to an embodiment of the present invention.

[0019] Reference numerals: 100, Sample loading groove; 200, Oil passage; 210, First oil port; 220, Second oil port; 300, Air passage; 400, Mounting groove; 500, Circuit board groove; 510, Socket groove; 600, Viewing window. Detailed Implementation

[0020] Embodiments of the present invention are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0021] In the description of this invention, the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limiting this invention.

[0022] In the description of this invention, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0023] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0024] Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention. The various technical features of the present invention can be combined interactively without contradicting each other.

[0025] Reference Figure 1 and Figure 2 Firstly, embodiments of the present invention propose an electrical sorting chip that can simultaneously take into account the convenience of sample loading and sample changing.

[0026] The electro-sorting chip includes a chip body and a circuit board. The chip body includes a sample loading structure and a sorting structure. The sample loading structure is provided with a sample loading tank 100, an oil channel 200, and an air channel 300. The sample loading tank 100 is located above the sorting structure. A droplet inlet is provided on the upper surface of the sample loading structure. The droplet inlet is connected to the upper end of the sample loading tank 100. One end of the air channel 300 extends to the side wall of the sample loading structure, and the other end is connected to the sample loading tank 100. The lower end of the sample loading tank 100 is connected to the sorting structure. One end of the oil channel 200 extends to the side wall of the sample loading structure, and the other end is connected to the sorting structure.

[0027] The circuit board is integrated on the sample mounting structure and electrically connected to the sorting structure. The sample mounting structure is provided with a circuit board groove 500 for mounting the circuit board. The sample mounting structure is also provided with wire-through holes to facilitate the electrical connection between the circuit board and the sorting structure. The wires led out from the circuit board located in the circuit board groove 500 can be soldered to the sorting structure through the wire-through holes to achieve conductive connection.

[0028] It is understandable that the sorting structure is equipped with sorting channels and electrodes, and the circuit board is electrically connected to the electrodes, enabling the electrodes to provide sorting voltage to the sorting channels.

[0029] During sample loading, the droplet suspension is added to the sample loading tank 100 through the droplet inlet, and driving air pressure is applied to the sample loading tank 100 through the air channel 300, causing the droplet suspension to be pressed down into the sorting structure below. Because a direct downward pressure method is used for droplet loading, excellent droplet condition can be ensured, such as... Figure 3 It is understandable that traditional centrifuge tubes are compressed upwards by air pressure and require sample addition via connecting tubes, resulting in a long injection time for the droplets, which can lead to abnormal droplet states. In this embodiment, however, the droplets are directly compressed downwards by full air pressure, which can shorten the droplet injection time, avoid abnormal droplet injection due to prolonged injection, and also take into account the convenience of air pressure sample loading and changing.

[0030] Understandably, since both the oil passage 200 and the air passage 300 extend to the side wall of the sample structure and the oil phase is external, when replacing the electric separation chip, the oil pipe can be directly connected through the opening of the oil passage 200 in the side wall of the sample structure. The air pressure supply structure can also be connected through the opening of the air passage 300 in the side wall of the sample structure.

[0031] In some embodiments, the oil channel 200 is provided with two oil channels, which extend to the side wall of the sample structure and form a first oil port 210 and a second oil port 220 on the side wall of the sample structure, respectively. The oil pipe can be directly connected to the electric sorting chip through the first oil port 210 and the second oil port 220.

[0032] In some embodiments, the sample-mounting structure is 3D printed using a transparent material, while the sorting structure is made of polydimethylsiloxane (PDMS). The chip body also includes a bonding layer, through which the sample-mounting structure and the sorting structure are connected. 3D printing technology allows for the fabrication of complex flow channels in the sample-mounting structure, such as the sample-mounting groove 100, oil channel 200, and air channel 300, which cannot be fabricated using conventional injection molding or machining. Furthermore, by printing the sample-mounting structure and then installing the sorting structure below it, the replacement of the sorting structure can be easily facilitated.

[0033] In some embodiments, the bonding layer is double-sided adhesive. The sorting structure made of PDMS is bonded to one side of the double-sided adhesive, while the other side of the double-sided adhesive is bonded to the sample loading structure made of 3D printing. Directly connecting the sample loading structure and the sorting structure with double-sided adhesive is simple to operate and inexpensive.

[0034] In other embodiments, the bonding layer is a glass plate. The sorting structure made of PDMS is bonded to one side of the glass plate, while the other side of the glass plate is bonded to the sample loading structure. Specifically, the sample loading structure can be achieved by double-sided / UV bonding the glass plate. Using a glass plate as the bonding layer reduces the risk of leakage after the sample loading structure and the sorting structure are connected, thus improving safety.

[0035] It is understandable that at least two through holes need to be provided on the glass plate so that the lower end of the sample loading tank 100 and the oil channel 200 can be connected to the sorting channel of the sorting structure through the through holes. It is also understandable that glass has strong high temperature resistance, and the through holes can be made by laser drilling.

[0036] In some embodiments, the lower end face of the sample structure is provided with a mounting groove 400 for mounting a sorting structure. The lower end of the sample groove 100 and the oil passage 200 are respectively connected to the mounting groove 400. During installation, the bonding layer is connected in the mounting groove 400.

[0037] In some embodiments, the electrical sorting chip further includes a power supply structure disposed on the sample structure and connected to the circuit board. It is understood that the power supply structure is used to connect to the power supply of a peripheral device, thereby supplying power to the circuit board.

[0038] The peripheral power supply can provide power to the circuit board of the electrical sorting chip through methods such as plug-in type, magnetic spring pin type, and direct contact.

[0039] In some embodiments, the power supply structure is a power supply socket. Specifically, a socket groove 510 for installing the power supply socket is provided on the upper sample structure. The socket groove 510 is located on the side of the circuit board groove 500. The power supply socket is integrated into the upper sample structure through the socket groove 510. The power plug can be directly inserted into the power supply socket and electrically connected to the circuit board located in the circuit board groove 500 to realize plug-in power supply.

[0040] In other embodiments, the power supply structure is a magnetic spring-loaded pin connector, integrated into the side wall of the sample structure. The magnetic spring-loaded pin connector consists of a spring pin, a magnet, and other components. When the power supply terminal approaches the connector, the magnetic attraction causes the spring pin to extend or retract, ensuring a tight fit between the pin and the contact point, forming a stable electrical connection and achieving magnetic spring-loaded pin-type power supply. By using this connector, not only is good electrical conduction efficiency between the power supply and the circuit board guaranteed, but the stability and durability of the connection are also improved.

[0041] In other embodiments, the power supply structure is a contact-type conductive element, which is integrated on the sample structure and connected to the circuit board. The power supply terminal can be directly conductively connected to the contact-type conductive element to achieve direct contact power supply.

[0042] In some embodiments, the electrical sorting chip further includes a pneumatic supply structure. The pneumatic supply structure is equipped with a supply plug, and one end of the air passage 300 extending to the side wall of the structure serves as the supply port. The supply plug and the supply port can be directly connected to achieve a direct-insertion pneumatic supply, or connected via a soft-plug adapter to achieve a soft-plug adapter-type pneumatic supply. Specifically, in the soft-plug adapter-type pneumatic supply scheme, the supply plug of the pneumatic supply structure is a rigid tube, and the supply port is a rigid hole. A soft-plug adapter with a through hole is provided inside the supply port, and the rigid tube is inserted into the through hole of the soft-plug adapter to achieve a sealed connection between the rigid tube and the rigid hole.

[0043] Understandably, using a soft-plug adapter for pneumatic supply can make the chip body more aesthetically pleasing.

[0044] In some embodiments, an annular plate is provided at the upper end of the sample tank 100. The annular plate gradually tilts downward from the upper wall of the sample tank 100 to the central axis of the sample tank 100. When the driving airflow enters the sample tank 100 along the air passage 300, it can be guided by the annular plate. The driving airflow applies downward pressure to the liquid in the sample tank 100, thereby driving the droplets to be pressed down into the sorting structure.

[0045] It is understandable that, in the case of a group of magnetic beads inside the droplet, in some embodiments, the electro-sorting chip also includes a wire magnet. A magnetic groove is provided below the sample loading structure, and the wire magnet is placed in the magnetic groove and connected to the sorting structure. This enables the wire magnet to be directly attached to the sorting structure, so as to realize the wire pulling of the magnetic beads inside the droplet. That is, under the action of the wire magnet, the group of magnetic beads inside the droplet is pulled into a wire for signal enrichment and fluorescence detection.

[0046] In some embodiments, the sample loading structure is provided with a viewing window 600, which extends through the upper and lower surfaces of the sample loading structure and is located above the sorting structure, allowing operators to easily observe the state of the droplets entering the sorting structure. Especially for sample loading structures made of non-transparent materials, the viewing window 600 allows operators to observe and monitor the droplet sorting effect.

[0047] In some embodiments, the chip body also includes a packaging board, which is connected to the sample structure and covers the upper end of the viewing window 600. The packaging board is a colored glass component or a filter, which can reduce the interference of ambient light on the electrical sorting while making it easy for operators to observe.

[0048] The overall size of the electric sorting chip is determined by the sorting structure. The length, width and height of the electric sorting chip range from 2-20 cm, 0.5-10 cm and 0.1-5 cm, respectively.

[0049] It is understood that using the electro-sorting chip of this embodiment to electro-sort droplets enhances ease of use, eliminating the need for refueling each time or setting up an injection pump; only the operation of adding droplets is required.

[0050] Secondly, embodiments of the present invention provide a method for fabricating an electrical sorting chip, which is applied to the electrical sorting chip proposed in any embodiment of the first aspect. The method for fabricating the electrical sorting chip includes steps S101, S102, and S103, as described above. Figure 4 .

[0051] Step S101: The sample loading structure is fabricated using 3D printing. In some embodiments, the sample loading structure is made of a transparent material to facilitate subsequent observation of droplet loading. It is understood that 3D printing technology can be used to fabricate complex flow channels in the sample loading structure, such as the sample loading groove 100, oil channel 200, and gas channel 300.

[0052] Step S102: Assemble the sorting structure below the sample structure.

[0053] It is understandable that the sample loading structure is installed above the sorting structure, and the lower end of the sample loading tank 100 is connected to the sorting channel of the sorting structure. The droplets entering from the sample loading tank 100 can be directly pressed down into the sorting structure under the action of air pressure, which shortens the injection time and avoids abnormal droplet states.

[0054] In some embodiments, the sorting structure is bonded to one side of a double-sided adhesive tape, while the other side of the double-sided adhesive tape is bonded to a 3D-printed sample structure. In other embodiments, the sorting structure is bonded to one side of a glass plate, while the other side of the glass plate is bonded to the sample structure; specifically, the sample structure can be made by double-sided / UV adhesive bonding of the glass plate.

[0055] In some embodiments, the sorting structure is made of PDMS material.

[0056] Step S103 involves mounting the circuit board onto the sample loading structure and soldering it to the sorting structure. This step integrates the circuit board, sorting structure, and sample loading structure, making it more convenient for users.

[0057] Thirdly, the embodiments of the present invention also propose an electrical sorting method. By applying the electrical sorting chip proposed in any of the embodiments of the first aspect to electrically sort droplets, the abnormal state of the droplets due to prolonged injection can be avoided, and the entire electrical sorting operation is simple. The electrical sorting method includes steps S201, S202, and S203, as described above. Figure 5 .

[0058] Step S201: Add the droplet suspension into the sample loading tank 100 from the droplet inlet.

[0059] In step S202, driving air pressure is applied to the air channel 300 to press the droplet suspension in the sample loading tank 100 down into the sorting structure. By applying driving air pressure to the sample loading tank 100 through the air channel 300, the droplet suspension is pressed down into the sorting structure located below. Because a direct downward pressure method is used for droplet loading, excellent droplet condition can be ensured.

[0060] Understandably, after sample loading, the droplet inlet needs to be covered, and driving air pressure is applied into the sample loading tank 100 to force the droplet suspension downwards into the sorting structure. Understandably, the droplet inlet can be covered with a cap or sealed with tape.

[0061] Understandably, the driving air pressure can be supplied via a direct-plug supply or a soft-plug transfer.

[0062] In step S203, when the droplets enter the sorting structure, power is supplied to the sorting structure via the circuit board. The circuit board supplies power to the sorting structure, providing a sorting voltage for the droplets on the sorting structure, thereby achieving electrical sorting of the droplets.

[0063] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. An electrical sorting chip, characterized in that, include: The chip body includes a sample loading structure and a sorting structure. The sample loading structure is provided with a sample loading groove, an oil channel, and an air channel. The sample loading groove is located above the sorting structure. The upper surface of the sample loading structure is provided with a droplet inlet, which is connected to the upper end of the sample loading groove. One end of the air channel extends to the side wall of the sample loading structure, and the other end is connected to the sample loading groove. The lower end of the sample loading groove is connected to the sorting structure. One end of the oil channel extends to the side wall of the sample loading structure, and the other end is connected to the sorting structure. A circuit board is disposed on the sample-upper structure and electrically connected to the sorting structure.

2. The electrical sorting chip according to claim 1, characterized in that, The sample loading structure is a 3D printed part made of transparent material; the sorting structure is a part made of polydimethylsiloxane material; the chip body also includes a bonding layer, and the sample loading structure and the sorting structure are connected through the bonding layer.

3. The electrical sorting chip according to claim 2, characterized in that, The bonding layer is double-sided adhesive, the sorting structure is bonded to one side of the double-sided adhesive, and the other side of the double-sided adhesive is bonded to the sample loading structure; or the bonding layer is a glass plate, the sorting structure is bonded to one side of the glass plate, and the other side of the glass plate is bonded to the sample loading structure, the glass plate is provided with at least two through holes, and the lower end of the sample loading groove and the oil passage are respectively connected to the sorting structure through the through holes.

4. The electrical sorting chip according to claim 1, characterized in that, The electrical sorting chip also includes a power supply structure, which is disposed on the sample structure and connected to the circuit board. The power supply structure is used to connect to an external power supply, and the power supply structure is one of a power supply socket, a magnetic spring pin connector, or a contact conductive component.

5. The electrical sorting chip according to claim 1, characterized in that, The electrical sorting chip also includes a pneumatic supply structure, which is equipped with a pneumatic supply plug. The end of the air passage extending to the side wall of the sample loading structure is a pneumatic supply port. The pneumatic supply plug is directly connected to the pneumatic supply port or is connected to the pneumatic supply port through a soft plug adapter.

6. The electrical sorting chip according to claim 1, characterized in that, The electrical sorting chip also includes a wire magnet. A magnet groove is provided below the sample loading structure. The wire magnet is located in the magnet groove and connected to the sorting structure.

7. The electrical sorting chip according to claim 1, characterized in that, The sample loading structure is provided with a viewing window that extends through the upper and lower surfaces of the sample loading structure and is located above the sorting structure.

8. The electrical sorting chip according to claim 7, characterized in that, The chip body also includes a packaging plate, which is connected to the sample mounting structure and covers the upper end of the window. The packaging plate is a colored glass component or a filter.

9. A method for fabricating an electrical sorting chip, characterized in that, Applied to the electrical sorting chip as described in any one of claims 1 to 8, the method for preparing the electrical sorting chip includes the following steps: The sample structure was fabricated using 3D printing. The sorting structure is assembled below the sample loading structure; The circuit board is mounted on the sample loading structure and soldered to the sorting structure.

10. An electrical separation method, characterized in that, The electrical sorting method, using the electrical sorting chip as described in any one of claims 1 to 8, comprises the following steps: The droplet suspension is added to the sample loading tank from the droplet inlet; A driving air pressure is applied to the air passage to press the droplet suspension in the sample loading tank down into the sorting structure; When a droplet enters the sorting structure, power is supplied to the sorting structure via a circuit board.

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