Electrotransfection chip

Abstract

The utility model provides a kind of electric conversion chip and its preparation method, the electric conversion chip includes first titanium electrode sheet, plastic support piece and second titanium electrode sheet;Plastic support piece has opposite first surface and second surface, plastic support piece is equipped with through slot that penetrates first surface and second surface, first titanium electrode sheet and second titanium electrode sheet are respectively attached in the first surface and second surface of plastic support piece, to block the opposite sides of through slot and form the electric conversion space with opening and for accommodating electric conversion liquid;When first titanium electrode sheet and second titanium electrode sheet contact with electric conversion liquid in electric conversion space and carry out electric conversion of cell in electric conversion liquid, the bottom surface of electric conversion space is inclined relative to horizontal plane arrangement.The utility model improves electric conversion performance, improves the utilization of electric conversion liquid.

Description

Technical Field

[0001] This invention belongs to the field of cell processing technology, and in particular relates to an electroporation chip. Background Technology

[0002] With the rapid development of medical technology, electroporation technology has become widely used and is an important method for non-viral cell transfection. Current electroporation chips typically consist of parallel aluminum electrodes and a rectangular cup structure. After adding the electroporation solution, the solution tends to form a concave surface at the top due to capillary action. This results in lower resistance at the corners of the surface, leading to insufficient cell voltage and decreased electroporation performance. Conversely, the voltage increases below the concave surface, potentially causing cell death. Furthermore, the relatively large bottom area makes it difficult to absorb small amounts of solution, leading to waste. Additionally, when aluminum is used as the anode, it is prone to reaction, causing electrode material wear and surface detachment. Aluminum ions generated during aluminum electrolysis and fine aluminum fragments contaminate the electroporation solution, reducing cell viability and ultimately resulting in poor electroporation performance.

[0003] Existing conventional cell electroporation devices utilize short-pulse voltages applied to the electrode surfaces of two rectangular, parallel, smooth aluminum substrates to create an electric field in the region between the electrode plates, enabling efficient electroporation delivery of exogenous substances into cells. With the development of technologies such as cell gene therapy, cell electroporation technology has gradually attracted researchers' attention due to its non-viral advantage. However, because the cells used for electroporation are usually human primary cells, limited in number and extremely precious, the amount and volume of cells used for electroporation are typically small, and high transfection performance is required. Existing traditional electroporation cups use a flat-bottomed structure with a large bottom area. When a small amount of electroporation system is added, the liquid height is small, making it difficult to aspirate. More importantly, capillary effects easily lead to an uneven liquid surface, forming a concave meniscus, ultimately resulting in uneven internal voltage distribution. At the concave corner of the liquid surface, the resistance is low, resulting in insufficient cell voltage and decreased electroporation performance; while below the concave meniscus, the voltage increases, making cells more prone to death. Therefore, for new technology applications such as cell gene therapy, which involve transfection of small numbers of cells and cell volumes, it is necessary to design new electrode structures that differ from conventional rectangular electrodes, or to improve traditional rectangular planar electrodes.

[0004] Meanwhile, although aluminum plates or aluminum-based electrodes can effectively open cell membranes and achieve a certain efficiency in the delivery of exogenous substances, aluminum is chemically reactive and readily reacts with oxygen to form an aluminum oxide protective layer. This layer is easily dissolved in electroporation buffer containing Cl⁻, leading to continuous corrosion of the aluminum metal. When aluminum is used as the anode, it readily undergoes the reaction (Al → Al³⁺ + 3e⁻), resulting in electrode material wear and surface detachment. The aluminum ions (Al³⁺) generated by aluminum electrolysis and the detached fine aluminum fragments contaminate the electroporation buffer and reduce cell viability, resulting in poor final electroporation performance. Furthermore, smooth aluminum-based electrodes require a high voltage to effectively open cell membranes, placing high demands on the power supply. Furthermore, although the transfection rate can be improved by sacrificing cell viability after transfection, or vice versa, by adjusting the electrical parameters applied to the two electrodes, such as voltage, pulse width, number of pulses, and pulse interval, the electrotransfection performance will still be lower than the optimal result. Therefore, how to achieve optimal electrotransfection results through structural and material design to meet the application scenarios and needs of cell and gene indicators is a key problem to be solved in this field. Summary of the Invention

[0005] This invention addresses the technical problems of low electro-spinning performance and low utilization rate of electro-spinning fluid in existing electro-spinning chips by providing an electro-spinning chip and its preparation method.

[0006] In view of the above technical problems, this utility model provides an electroporation chip, including a first titanium electrode, a plastic support, and a second titanium electrode. The plastic support has a first surface and a second surface opposite to each other, and a through groove penetrating the first surface and the second surface is provided on the plastic support. The first titanium electrode and the second titanium electrode are respectively attached to the first surface and the second surface of the plastic support to block the opposite sides of the through groove and form an electroporation space with an opening for containing electroporation solution. When the first titanium electrode and the second titanium electrode are in contact with the electroporation solution in the electroporation space and perform electroporation on the cells in the electroporation solution, the bottom surface of the electroporation space is inclined relative to the horizontal plane.

[0007] Optionally, the first titanium electrode sheet and the second titanium electrode sheet are arranged in parallel; the through groove includes a first inner sidewall, a second inner sidewall, and a bottom surface, all perpendicular to the first titanium electrode sheet; the bottom surface is connected between the first inner sidewall and the second inner sidewall; the opening of the electro-rotation space is arranged opposite to the bottom surface; When performing electroporation on cells in the electroporation solution, the first titanium electrode is perpendicular to the horizontal plane, and the bottom surface is set at a preset tilt angle to the horizontal plane.

[0008] Optionally, the preset tilt angle is 0-45 degrees; or / and When performing electroporation on cells in the electroporation solution, both the first inner wall and the second inner wall are perpendicular to the horizontal plane.

[0009] Optionally, the ratio between the depth and width of the electro-rotation space ranges from 1:1 to 5:1; wherein, the depth of the electro-rotation space refers to the distance between the opening and a preset point on the bottom surface, and the preset point is any point on the bottom surface; the width of the electro-rotation space refers to the distance between the first inner sidewall and the second inner sidewall when both the first titanium electrode sheet and the second titanium electrode sheet are attached to the plastic support.

[0010] Optionally, a first connecting portion is provided on the first surface around the through groove, and the first titanium electrode sheet is sealed and bonded to the first surface of the plastic support through the first connecting portion; The second surface is provided with a second connecting portion surrounding the through groove, and the second titanium electrode sheet is sealed and bonded to the second surface of the plastic support through the second connecting portion.

[0011] Optionally, the first connecting portion is a first adhesive guide groove for guiding adhesive into the first surface and the first titanium electrode sheet; the second connecting portion is a second adhesive guide groove for guiding adhesive into the second surface and the second titanium electrode sheet.

[0012] Optionally, the first titanium electrode sheet is provided with at least two first positioning holes; the plastic support is provided with at least two second positioning holes opposite to each of the first positioning holes; the second titanium electrode sheet is provided with at least two third positioning holes opposite to each of the second positioning holes; the electro-rotation chip further includes at least two first positioning posts, each of which is used to sequentially pass through the first positioning hole, the second positioning hole, and the third positioning hole to position the first titanium electrode sheet, the plastic support, and the second titanium electrode sheet.

[0013] Optionally, the first titanium electrode sheet is provided with at least two fourth positioning holes; the second titanium electrode sheet is provided with at least two fifth positioning holes; the plastic support member is provided with second positioning posts and third positioning posts on opposite sides respectively; the second positioning posts are arranged opposite to the fourth positioning holes so as to position the first titanium electrode sheet and the plastic support member when each of the second positioning posts passes through the fourth positioning holes in a corresponding manner; the third positioning posts are arranged opposite to the fifth positioning holes so as to position the second titanium electrode sheet and the plastic support member when each of the third positioning posts passes through the fifth positioning holes in a corresponding manner.

[0014] Optionally, the first titanium electrode sheet includes a first titanium substrate and a first metal coating on the first titanium substrate; the second titanium electrode sheet includes a second titanium substrate and a second metal coating on the second titanium substrate.

[0015] Optionally, the plastic support is a modified part with low surface properties; or / and The outer surface of the plastic support is covered with a hydrophobic layer; or / and The surface roughness of the first titanium electrode sheet facing the first surface ranges from Ra1.0 to Ra3.0; the surface roughness of the second titanium electrode sheet facing the second surface ranges from Ra1.0 to Ra3.0.

[0016] The electroporation chip provided by this utility model includes a first titanium electrode, a plastic support, and a second titanium electrode. The plastic support has a first surface and a second surface facing each other, and a through groove penetrating the first surface and the second surface is provided on the plastic support. The first titanium electrode and the second titanium electrode are respectively attached to the first surface and the second surface of the plastic support to block the opposite sides of the through groove and form an electroporation space with an opening for containing electroporation solution. When the first titanium electrode and the second titanium electrode are in contact with the electroporation solution in the electroporation space and perform electroporation on the cells in the electroporation solution, the bottom surface of the electroporation space is inclined relative to the horizontal plane.

[0017] In this invention, by using a first titanium electrode and a second titanium electrode to electrotransfer cells in the electrotransfer solution, the electrotransfer process is prevented from detaching and electrolyzing, thus avoiding contamination of the electrotransfer solution and improving electrotransfer performance. Simultaneously, the first and second titanium electrode plates also reduce capillary action on the electrotransfer solution, decreasing cell death during electrotransfer and further improving performance. Furthermore, the inclined bottom surface of the electrotransfer space relative to the horizontal plane facilitates the aspiration of small amounts of electrotransfer solution, improving its utilization rate. Attached Figure Description

[0018] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0019] Figure 1 This is a schematic diagram of the structure of an electro-electric converter chip provided in an embodiment of this utility model.

[0020] Figure 2 This is a schematic diagram of the structure of an electro-electric converter chip provided in another embodiment of this utility model.

[0021] Figure 3 This is a schematic diagram of the structure of an electro-electric converter chip provided in another embodiment of the present invention.

[0022] Figure 4This is a schematic diagram illustrating the experimental results of plasmid electroporation of Jurkat cells according to an embodiment of this utility model.

[0023] Figure 5 This is a schematic diagram of an electrical transfer space with different depth-to-width ratios provided in an embodiment of the present invention.

[0024] Figure 6 This is a schematic diagram showing the experimental results of transfection rate and viability of electroporation spaces with different depth-to-width ratios provided in an embodiment of this utility model.

[0025] Figure 7 This is a flowchart of a method for preparing an electro-particle chip according to an embodiment of the present invention.

[0026] Figure 8 This is a schematic diagram of the fabrication process of the electro-electric chip provided in one embodiment of the present invention.

[0027] Figure 9 This is a schematic diagram of an electro-rotating chip with different preset tilt angles provided in an embodiment of the present invention.

[0028] The reference numerals in the accompanying drawings are as follows: 100, First titanium electrode sheet; 110, First positioning hole; 200, Plastic support; 210, Through groove; 211, First inner sidewall; 212, Second inner sidewall; 213, Bottom surface; 220, First connecting part; 230, Second connecting part; 240, Second positioning hole; 300, Second titanium electrode sheet; 310, Third positioning hole; 400, Electro-rotation space; 410, Opening; 500, First positioning post. Detailed Implementation

[0029] To make the technical problems solved, technical solutions, and beneficial effects of this utility model clearer, the present utility model will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain this utility model and are not intended to limit this utility model.

[0030] like Figures 1 to 3 As shown, one embodiment of this utility model provides an electro-spinning chip, including a first titanium electrode 100, a plastic support 200, and a second titanium electrode 300; the plastic support 200 has opposing first and second surfaces, and a through groove 210 penetrating the first and second surfaces is provided on the plastic support 200; the first titanium electrode 100 and the second titanium electrode 300 are respectively attached to the first and second surfaces of the plastic support 200 to block the opposing sides of the through groove 210 and form an electro-spinning space 400 with an opening 410 for containing electro-spinning fluid; as shown Figure 3As shown, when the first titanium electrode 100 and the second titanium electrode 300 are in contact with the electrotransfer solution in the electrotransfer space 400 and perform electrotransfer staining on the cells in the electrotransfer solution, the bottom surface 213 of the electrotransfer space 400 is inclined relative to the horizontal plane.

[0031] Understandably, the side of the first titanium electrode 100 facing the first surface or the side of the second titanium electrode 300 facing the second surface has a certain roughness. This roughness can be set according to actual conditions to increase the contact area between the first titanium electrode 100 or the second titanium electrode 300 and the electroplating fluid, while simultaneously improving the hydrophobicity of the first titanium electrode 100 or the second titanium electrode 300 and reducing the capillary action on the electroplating fluid, thereby forming a nearly horizontal liquid surface. The thickness of the plastic support 200, i.e., the distance between the first titanium electrode 100 and the second titanium electrode 300, can be set according to actual conditions, for example, 2mm, 4mm, or 6mm. The opening 410 of the electroplating space 400 can be set at different positions in the electroplating space 400 according to actual conditions (set by adjusting the position of the slot opening 210), as long as the electroplating space 400 can accommodate the required electroplating fluid volume.

[0032] This invention provides a strategy for adjusting the design of the electrotransfer chamber structure while maintaining optimal electrotransfer results. By adjusting the depth-to-width ratio of the electrotransfer chamber, a higher transfection rate or cell viability can be obtained while maintaining optimal cell electrotransfer results. This allows for achieving higher transfection rates or cell viability after cell electrotransfer, based on actual design requirements. Specifically, the ratio between the depth and width of the electrotransfer space 400 can be set according to actual conditions. The depth of the electrotransfer space 400 refers to the distance between the opening 410 and a preset point on the bottom surface 213, where the preset point is any point on the bottom surface 213. The width of the electrotransfer space 400 refers to the distance between the first inner wall 211 and the second inner wall 212 when both the first titanium electrode 100 and the second titanium electrode 300 are attached to the plastic support 200. Specifically, when the opening 410 is parallel to the horizontal plane, the depth of the electroporation space 400 refers to the distance between the top surface of the opening 410 and a preset point on the bottom surface 213. When the opening 410 is not parallel to the horizontal plane, the depth of the electroporation space 400 refers to the distance between the lowest point of the opening 410 and a preset point on the bottom surface 213. Since the bottom surface 213 is inclined relative to the horizontal plane, the depth of the electroporation space 400 changes depending on the position of the preset point on the bottom surface 213, but the range of depth variation of the electroporation space 400 remains within a predetermined range. For example, the ratio of the depth to width of the electroporation space 400 corresponding to different preset points is always within the range of 1:1 to 5:1, thereby achieving a higher transfection rate or higher cell viability while maintaining optimal cell electroporation results.

[0033] The first titanium electrode 100 or the second titanium electrode 300 can have its conductivity and biocompatibility improved by coating. The coating can be one or more of titanium plating with gold, platinum, copper, or silver. The plastic support 200 can be made of one or more of polymethyl methacrylate, polycarbonate, polypropylene, polystyrene, polyphenylene sulfide, polyethylene, polyethylene terephthalate, or cyclic olefin copolymers. In this embodiment, a first metal coating and a second metal coating are respectively applied to the first titanium substrate of the first titanium electrode 100 and the second titanium substrate of the second titanium electrode 300, which can increase the electroporation contact area and reduce contact resistance. The tilt angle of the bottom surface 213 of the electroporation space 400 relative to the horizontal plane can be set according to actual conditions, thereby facilitating the extraction of small amounts of sample after electroporation, reducing cell loss during experimental operations, and improving cell recovery rate.

[0034] like Figure 4As shown, in the plasmid electroporation experiment on Jurkat cells, chip A is the electroporation chip of this invention, using a first titanium electrode 100, a plastic support 200, and a second titanium electrode 300; while chip B uses electrodes made of smooth aluminum. The plasmid is pre-modified with enhanced green fluorescent protein (EGFP). Specifically, the cultured cells are replaced with the same electroporation buffer, and the cell density is adjusted to 1 x 10⁻⁶ cells / year. 7 or / and mL -1 Plasmids were added to the electroporation buffer to achieve a final concentration of 20 μg / mL, forming the cell sample for electroporation. Equal volumes of cell samples were extracted and added to different chips (A and B), and electroporation parameters of 320V, 2ms pulse width, and one pulse were applied. After electroporation, the cell samples were quickly transferred to fresh whole culture medium and incubated statically in a cell culture incubator (37℃, constant 5% CO2 concentration). After 48 hours, cells successfully transfected with the plasmid emitted green fluorescence at 507nm after excitation with 494nm light. Cell viability was measured for different cell samples, and flow cytometry was used to quantify the transfected cells. Figure 4 The experimental results shown indicate that using chip A, with its first titanium electrode 100, plastic support 200, and second titanium electrode 300 for cell transfection, the cell transfection rate, viability, and recovery rate are all higher than those of chip B, which uses smooth aluminum electrode sheets.

[0035] In the above embodiments of this invention, by setting a first titanium electrode 100 and a second titanium electrode 300 to perform electrotransfection of cells in the electrotransfection solution, the electrotransfection of cells is prevented from occurring due to electrode detachment and electrolysis during the electrotransfection process, thus avoiding contamination of the electrotransfection solution and improving electrotransfection performance. Simultaneously, the first titanium electrode 100 and the second titanium electrode 300 can also reduce capillary action on the electrotransfection solution, reducing cell death during electrotransfection and further improving electrotransfection performance. Furthermore, the inclined arrangement of the bottom surface 213 of the electrotransfection space 400 relative to the horizontal plane facilitates the aspiration of small amounts of electrotransfection solution, improving the utilization rate of the electrotransfection solution.

[0036] like Figures 1 to 3As shown, in one embodiment, the first titanium electrode 100 and the second titanium electrode 300 are arranged in parallel; the through groove 210 includes a first inner sidewall 211, a second inner sidewall 212, and a bottom surface 213, all perpendicular to the first titanium electrode 100; the bottom surface 213 is connected between the first inner sidewall 211 and the second inner sidewall 212; the opening 410 of the electroporation space 400 is arranged opposite to the bottom surface 213; when performing electroporation on cells in the electroporation solution, the first titanium electrode 100 is perpendicular to the horizontal plane, and the bottom surface 213 is set at a preset tilt angle to the horizontal plane. Understandably, the first titanium electrode 100 and the second titanium electrode 300 are arranged in parallel, and the first inner sidewall 211 and the second inner sidewall 212 are both arranged vertically between the first titanium electrode 100 and the second titanium electrode 300. This allows the cells in the electrotransfer solution between the first titanium electrode 100 and the second titanium electrode 300 to receive equal electrotransfection voltage, thereby increasing the cell porosity and allowing more exogenous substances to be delivered into the cells for transfection, thus improving the transfection rate.

[0037] In one embodiment, the preset tilt angle is 0-45 degrees. Understandably, within the electrotransfer space 400 where the bottom surface 213 is tilted, more cells experience a uniform electric field, resulting in a higher cell porosity and the delivery of more exogenous substances into the cells for electrotransfection, leading to a lower cell viability. Conversely, in the electrotransfer space 400 where the bottom surface 213 is not tilted, capillary action causes more cells to be located in the low partial pressure region of the upper solution, meaning the cells experience a lower electric field strength, resulting in a lower transfection rate and higher viability in this portion of the cells. Meanwhile, the high partial pressure region of the upper solution has a smaller solution volume compared to the low partial pressure region, therefore the overall transfection rate of the sample is lower, and the viability is higher. Figure 2 , Figure 3 and Figure 9 ( Figure 9 As shown in the figure, the bottom tilt angle is the preset tilt angle. In one embodiment, the preset tilt angle includes, but is not limited to, 10 degrees, 20 degrees or 30 degrees.

[0038] like Figure 3As shown, in one embodiment, during electrotransfection of cells in the electrotransfer solution, both the first inner sidewall 211 and the second inner sidewall 212 are perpendicular to the horizontal plane. Understandably, since the first inner sidewall 211 and the second inner sidewall 212 are also perpendicular to the first titanium electrode 100, which is perpendicular to the horizontal plane, it can be known that the first inner sidewall 211 and the second inner sidewall 212 are also arranged parallel to each other, forming a rectangular space between the first titanium electrode 100, the first inner sidewall 211, the second titanium electrode 300, and the second inner sidewall 212. At this time, the first inner sidewall 211 and the second inner sidewall 212 are inclined to the bottom surface 213. Furthermore, the cells in the electrotransfer solution within the rectangular space receive a uniform electrotransfection voltage, resulting in a higher cell porosity, allowing more exogenous substances to be delivered into the cells for transfection, thus improving the transfection rate.

[0039] In one embodiment, the ratio of the depth to the width of the electroporation space 400 ranges from 1:1 to 5:1; wherein, the depth of the electroporation space 400 refers to the distance between the opening 410 and a preset point on the bottom surface 213, the preset point being any point on the bottom surface 213; the width of the electroporation space 400 refers to the distance between the first inner sidewall 211 and the second inner sidewall 212 when both the first titanium electrode 100 and the second titanium electrode 300 are attached to the plastic support 200. It can be understood that by setting the ratio of the depth to the width of the electroporation space 400 within the range of 1:1 to 5:1, a higher transfection rate or higher cell viability can be obtained while maintaining optimal cell electroporation results.

[0040] In this embodiment, schematic diagrams of electrical transfer spaces 400 with different depth-to-width ratios are shown below. Figure 5 As shown. The experimental results for its transfection rate and viability are as follows. Figure 6As shown, increasing the depth-to-width ratio from 1:2 to 1:5 resulted in a decrease in cell transfection rate from 76.35% to 60.47%, while viability increased from 61.29% to 76.23%, with the product of transfection rate and viability remaining between 45% and 48%. Therefore, if a higher transfection rate is desired after electrotransfection, an electrotransfection chamber with a lower depth-to-width ratio (e.g., 400) can be chosen; conversely, if a higher viability is desired, an electrotransfection chamber with a higher depth-to-width ratio (e.g., 400) can be chosen. However, the design of a high depth-to-width ratio needs to consider the operability and convenience of the experiment; a ratio between 1:1 and 5:1 is generally suitable. An excessively high ratio may prevent effective collection of samples from the bottom of the chamber, reducing sample recovery rate. In this embodiment, an improved strategy for the electroporation space of a traditional rectangular plane is provided. By optimizing the depth-to-width ratio of the electroporation space 400, users can meet specific application requirements, such as the need for a high cell transfection rate or cell viability, while maintaining the best electroporation results.

[0041] like Figure 2 and Figure 3 As shown, in one embodiment, a first connecting portion 220 is provided on the first surface surrounding the through groove 210, and the first titanium electrode 100 is sealed and bonded to the first surface of the plastic support 200 through the first connecting portion 220; a second connecting portion 230 is provided on the second surface surrounding the through groove 210, and the second titanium electrode 300 is sealed and bonded to the second surface of the plastic support 200 through the second connecting portion 230. It can be understood that both the first connecting portion 220 and the second connecting portion 230 are provided around the through groove 210, thereby facilitating the sealing of the area around the through groove 210, thus forming an electro-spreading space 400 that is sealed except for the opening 410, preventing leakage of the electro-spreading fluid and causing pollution or waste. The structure of the first connecting portion 220 or the second connecting portion 230 can be configured according to actual conditions, such as a guide groove or a protruding structure, as long as it can ensure that the area around the through groove 210 is sealed, thereby forming an electro-spreading space 400 that is sealed except for the opening 410.

[0042] like Figure 2 and Figure 3As shown, in one embodiment, the first connecting portion 220 is a first adhesive guide groove for guiding adhesive into the first surface and the first titanium electrode sheet 100; the second connecting portion 230 is a second adhesive guide groove for guiding adhesive into the second surface and the second titanium electrode sheet 300. After the adhesive is introduced through the first and second adhesive guide grooves, it is convenient to firmly bond the first surface and the first titanium electrode sheet 100, as well as the second surface and the second titanium electrode sheet 300, thereby forming an electro-rotation space 400 that is sealed except for the opening 410.

[0043] like Figure 8 As shown, in one embodiment, the first titanium electrode sheet 100 is provided with at least two first positioning holes 110; the plastic support 200 is provided with at least two second positioning holes 240 opposite to each of the first positioning holes 110; the second titanium electrode sheet 300 is provided with at least two third positioning holes 310 opposite to each of the second positioning holes 240; the electro-rotating chip further includes at least two first positioning posts 500, each of the first positioning posts 500 being used to sequentially pass through the first positioning hole 110, the second positioning hole 240 and the third positioning hole 310 to position the first titanium electrode sheet 100, the plastic support 200 and the second titanium electrode sheet 300. Understandably, during the fabrication of the electro-particle chip, the first positioning post 500 can be first inserted between the first positioning hole 110 and the second positioning hole 240 to position the first titanium electrode 100 and the plastic support 200, thereby facilitating the bonding of the first titanium electrode 100 and the plastic support 200. Then, the third positioning hole 310 can be inserted into the end of the first positioning post 500 opposite to the first positioning hole 110 to position the second titanium electrode 300 and the plastic support 200, thereby facilitating the bonding of the second titanium electrode 300 and the plastic support 200. In another embodiment, the first positioning post 500 can also be directly passed through the first positioning hole 110, the second positioning hole 240, and the third positioning hole 310 in one go, thereby completing the positioning and bonding of the first titanium electrode 100, the plastic support 200, and the second titanium electrode 300 in one operation. In this embodiment, after the positioning and bonding between the first titanium electrode sheet 100, the plastic support 200 and the second titanium electrode sheet 300 are completed, all the first positioning posts 500 can be removed, thereby making the overall weight of the electro-spinning chip lighter and easier to operate. In addition, the first positioning posts 500 can be reused in electro-spinning chips of the same specification, reducing costs.

[0044] In one embodiment, the first titanium electrode sheet 100 is provided with at least two fourth positioning holes; the second titanium electrode sheet 300 is provided with at least two fifth positioning holes; the plastic support 200 is provided with second positioning posts and third positioning posts on opposite sides respectively; the second positioning posts are arranged opposite to the fourth positioning holes so as to position the first titanium electrode sheet 100 and the plastic support 200 when each of the second positioning posts passes through the fourth positioning holes in a corresponding manner; the third positioning posts are arranged opposite to the fifth positioning holes so as to position the second titanium electrode sheet 300 and the plastic support 200 when each of the third positioning posts passes through the fifth positioning holes in a corresponding manner. Understandably, during the fabrication of the electro-particle chip, the second positioning post can be first inserted into the fourth positioning hole to position the first titanium electrode 100 and the plastic support 200, thereby facilitating the bonding between them. Then, the third positioning post 500 can be inserted into the fifth positioning hole to position the second titanium electrode 300 and the plastic support 200, further facilitating the bonding between them. The order of these steps can also be adjusted, i.e., the third positioning post 500 can be inserted into the fifth positioning hole first, followed by the second positioning post into the fourth positioning hole. In this embodiment, the plastic support 200, the second positioning post, and the third positioning post can be an integrally formed structure, thus simplifying the fabrication process.

[0045] In one embodiment, the first titanium electrode 100 includes a first titanium substrate and a first metal plating layer coated on the first titanium substrate; the second titanium electrode 300 includes a second titanium substrate and a second metal plating layer coated on the second titanium substrate. Understandably, the first metal plating layer is used to improve the conductivity and biocompatibility of the first titanium electrode 100, and the second metal plating layer is used to improve the conductivity and biocompatibility of the second titanium electrode 300. In this embodiment, the first titanium substrate of the first titanium electrode 100 and the second titanium substrate of the second titanium electrode 300 are respectively covered with the first metal plating layer and the second metal plating layer, which can increase the electro-transfer contact area and reduce the contact resistance. The first metal plating layer can be made of one or more of gold, platinum, copper, or silver. For example, when the first metal plating layer is made of gold, the first titanium electrode refers to a titanium electrode with a gold layer plated on the first titanium substrate; that is, the first titanium electrode is a titanium-plated gold electrode. Similarly, the second metal plating layer can be made of one or more of gold, platinum, copper, or silver. For example, if the material of the second metal plating layer is silver, then the second titanium electrode sheet refers to a silver layer plated on the second titanium substrate, that is, the second titanium electrode sheet is a titanium-plated silver electrode sheet.

[0046] In one embodiment, the plastic support 200 is a modified component with low surface properties. Understandably, the plastic support 200 can be made of one or more of polymethyl methacrylate, polycarbonate, polypropylene, polystyrene, polyphenylene sulfide, polyethylene, polyethylene terephthalate, or cyclic olefin copolymers. The plastic support 200 can have its surface properties reduced through surface modification, thereby increasing its hydrophobicity, reducing the capillary effect of the plastic support 200 on the electroplating fluid, and increasing the effective electroplating area in the electroplating space 400.

[0047] Specifically, in the narrow electroporation chamber of a traditional electroporation cup, due to the surface capillary action of the cell suspension in the electroporation solution, the combined effects of adsorption (adhesion) between liquid and solid molecules and cohesion (cohesion) between liquid molecules cause the liquid to rise along the electrode wall at the contact point, forming a concave meniscus. This results in the actual liquid level in the central region of the electroporation chamber being lower than the liquid level near the electrode at the edge. In the above embodiments of this invention, surface modification of the plastic support reduces the electric field inhomogeneity caused by the capillary effect, maintaining a larger effective electroporation area. Simultaneously, it reduces the power requirements of the external power supply for electroporation and decreases cell damage under the same electrical parameters.

[0048] In one embodiment, the outer surface of the plastic support 200 is coated with a hydrophobic layer. In this embodiment, the surface hydrophobicity of the plastic support 200 is improved by coating it with a hydrophobic layer, thereby reducing the capillary effect of the plastic support 200 on the electro-spreading fluid and increasing the effective electro-spreading area in the electro-spreading space 400.

[0049] Furthermore, in this invention, the plastic support 200 can be surface modified by the following method to obtain a modified part with low surface properties: 1. Surface Coating / Immersion Coating: The plastic support 200 is immersed in a solution containing hydrophobic components (such as fluorinated siloxanes, long-chain alkyl silanes, or other hydrophobic polymers), and then cured by heating or ultraviolet light. Subsequently, the hydrophobic molecules adhere to the surface of the plastic support 200 through physical adsorption or chemical bonding, forming a low surface energy film, ultimately obtaining a modified part with low surface properties.

[0050] 2. Surface grafting: First, active free radicals are generated on the surface of the plastic support 200 by means of plasma or ultraviolet light. Then, the plastic support 200 is exposed to hydrophobic monomer (such as fluorinated acrylate) vapor or solution, so that these monomers polymerize on the surface of the plastic support 200 to "grow" hydrophobic polymer brushes. By precisely controlling the grafting density and chain length, the degree of hydrophobicity is customized to obtain a very uniform and stable modified surface, that is, a modified part with low surface properties is obtained.

[0051] In one embodiment, the surface roughness of the first titanium electrode 100 facing the first surface ranges from Ra1.0 to Ra3.0; the surface roughness of the second titanium electrode 300 facing the second surface ranges from Ra1.0 to Ra3.0. In this embodiment, the first titanium electrode 100 and the second titanium electrode 300 having surface roughnesses within the above-mentioned ranges can increase the contact area with the electroplating fluid while reducing the impedance of the first titanium electrode 100 and the second titanium electrode 300.

[0052] Understandably, the electrical connection in the electroporation process can be equivalent to a series circuit of an external power supply, an electroporation chip, and an electroporation solution, i.e., U = Uc + UR. Here, U is the external input voltage, Uc is the voltage division between the first titanium electrode 100 and the second titanium electrode 300, and UR is the voltage division of the electroporation solution. To ensure that the current applied to the electroporation solution remains consistent, the overall impedance of the series circuit decreases when the impedance of the first titanium electrode 100 and the second titanium electrode 300 is reduced. This reduces the required external input voltage and consequently the voltage demand on the external power supply. Furthermore, increasing the surface roughness of the first titanium electrode 100 and the second titanium electrode 300 can effectively reduce energy loss caused by the double-layer effect. Specifically, the contact state between the first titanium electrode 100 and the second titanium electrode 300 and the cells in the electroporation solution easily forms a double-layer capacitor (C0) on the surface of the first titanium electrode 100 or the second titanium electrode 300. d1 Based on the relationship between double-layer capacitance and electrode surface area, i.e. A is the electrode surface area. It is the vacuum permittivity; It is the relative permittivity; The thickness of the electric double layer is determined by increasing the surface roughness of the first titanium electrode 100 and the second titanium electrode 300, thereby increasing the capacitance of the electric double layer. Meanwhile, U... C =1 / jwC d1 XI, j is the imaginary unit, w is the angular frequency, I is the current, then U C With a reduced voltage divider, the total voltage requirement for external power supply equipment is also reduced.

[0053] like Figure 7 and Figure 8 As shown, one embodiment of this utility model also provides a method for preparing the electro-particle chip, including the following steps S100-S200: S100: UV-curable adhesive or thermosetting adhesive is dripped along a preset adhesive guide trajectory at a preset dispensing rate between the first surface of the plastic support 200 and the first titanium electrode 100, and between the second surface and the second titanium electrode 300. Understandably, the electro-spinning chip prepared in this method is the aforementioned electro-spinning chip. The specific structure and detailed preparation process of the electro-spinning chip can be referred to in the above embodiments and will not be repeated here. The preset adhesive guide trajectory can be determined by the specific setting of adhesive guide grooves or protrusion structures, etc. That is, if a first adhesive guide groove is provided on the first surface and a second adhesive guide groove is provided on the second surface, the first adhesive guide groove can be used to guide the adhesive (UV-curable adhesive or thermosetting adhesive) into the space between the first surface and the first titanium electrode 100, and the second adhesive guide groove can be used to guide the adhesive into the space between the second surface and the second titanium electrode 300, thereby preventing adhesive leakage and contamination of the electro-spinning space 400. In this case, the preset adhesive guide trajectory is along the direction of the first and second adhesive guide grooves. UV-curable adhesive or thermosetting adhesive is applied at a preset dispensing rate along a preset adhesive guide trajectory between the first surface and the first titanium electrode 100, and between the second surface and the second titanium electrode 300. This can be done simultaneously. The preset dispensing rate can be set according to actual needs and is not limited here.

[0054] In some embodiments, the steps of dripping UV-curable adhesive or thermosetting adhesive along a preset adhesive guide trajectory at a preset dispensing rate between the first surface and the first titanium electrode 100, and between the second surface and the second titanium electrode 300, can also be performed separately. In this case, refer to Figure 8 As shown, step S100 includes the following steps S101 and S102: S101. Apply UV-curable adhesive or thermosetting adhesive at a preset dispensing rate along a preset adhesive guide trajectory between the first surface of the plastic support 200 and the first titanium electrode sheet 100. S102. Apply UV-curable adhesive or thermosetting adhesive to the second surface of the plastic support 200 and between the second titanium electrode sheet 300 at a preset dispensing rate along a preset adhesive guide trajectory.

[0055] S200: After confirming the alignment of the first titanium electrode 100, the plastic support 200, and the second titanium electrode 300, the UV-curable adhesive is cured by UV irradiation for a first preset duration, or by thermosetting the adhesive in a heating oven for a second preset duration, to obtain the electro-rotation chip. The first and second preset durations can be set according to actual conditions. For example, the first preset duration can be 5-10 seconds, as long as the UV-curable adhesive is cured. The second preset duration can be set based on the curing time of the thermosetting adhesive, and can be equal to or different from the first preset duration, as long as the UV-curable adhesive is cured. Understandably, if ultraviolet-curing adhesive or thermosetting adhesive is simultaneously applied at a preset dispensing rate along a preset adhesive guide trajectory between the first surface and the first titanium electrode sheet 100, and between the second surface and the second titanium electrode sheet 300, then the ultraviolet-curing adhesive on both sides can be simultaneously cured by irradiating the first surface and the second surface with ultraviolet light for a first preset time, or the first surface and the second surface can be simultaneously heated in a heating oven for a second preset time to synchronously cure the thermosetting adhesive on both sides of the first surface and the second surface, thereby reducing the curing time.

[0056] Understandably, in such Figure 8 In the illustrated embodiment, if ultraviolet-curable adhesive is applied at a preset dispensing rate along a preset adhesive guide trajectory between the first surface and the first titanium electrode 100, and between the second surface and the second titanium electrode 300, respectively, the ultraviolet-curable adhesive or the thermosetting adhesive can be cured by irradiating with ultraviolet light for a first preset time or by curing with a heating oven for a second preset time. Specifically, after step S101, the following step is included: step S201, after determining that the first titanium electrode 100 and the plastic support 200 are aligned, the ultraviolet-curable adhesive is cured by irradiating with ultraviolet light for a first preset time.

[0057] The step S102 above includes: step S202, after determining that the plastic support 200 and the second titanium electrode sheet 300 are directly aligned, the ultraviolet curing adhesive is cured by ultraviolet light irradiation for a first preset duration, or the thermosetting adhesive is cured for a second preset duration by heating oven to obtain the electro-rotation chip.

[0058] The method for preparing an electroporation chip provided by this invention allows for electroporation of cells in the electroporation solution using a first titanium electrode 100 and a second titanium electrode 300. This avoids electrode detachment and electrolysis during the electroporation process, preventing contamination of the electroporation solution and improving electroporation performance. Simultaneously, the first and second titanium electrode 100 reduce capillary action on the electroporation solution, decreasing cell death during electroporation and further enhancing performance. Furthermore, the inclined bottom surface 213 of the electroporation space 400 relative to the horizontal plane facilitates the aspiration of small amounts of electroporation solution, improving its utilization rate.

[0059] Understandably, the method for preparing the electro-spinning chip provided by this utility model also includes methods for preparing other detailed structures in the electro-spinning chip, which can be referred to in the specific description of the electro-spinning chip, and will not be repeated here.

[0060] It should be understood that the sequence number of each step in the above embodiments does not imply the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of this utility model embodiment.

[0061] The above are merely embodiments of the electro-spinning chip and its preparation method of this utility model, and are not intended to limit this utility model. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this utility model should be included within the protection scope of this utility model.

Claims

1. An electro-electric converter chip, characterized in that, It includes a first titanium electrode sheet, a plastic support, and a second titanium electrode sheet; The plastic support has a first surface and a second surface opposite to each other. The plastic support is provided with a through groove that penetrates the first surface and the second surface. The first titanium electrode sheet and the second titanium electrode sheet are respectively attached to the first surface and the second surface of the plastic support to block the opposite sides of the through groove and form an electro-switching space with an opening for containing electro-switching fluid. When the first titanium electrode and the second titanium electrode are in contact with the electroporation solution in the electroporation space and perform electroporation on the cells in the electroporation solution, the bottom surface of the electroporation space is inclined relative to the horizontal plane.

2. The electro-electric converter chip according to claim 1, characterized in that, The first titanium electrode sheet and the second titanium electrode sheet are arranged in parallel; the through groove includes a first inner sidewall, a second inner sidewall, and a bottom surface, all perpendicular to the first titanium electrode sheet; the bottom surface is connected between the first inner sidewall and the second inner sidewall; the opening of the electro-rotation space is arranged opposite to the bottom surface; When performing electroporation on cells in the electroporation solution, the first titanium electrode is perpendicular to the horizontal plane, and the bottom surface is set at a preset tilt angle to the horizontal plane.

3. The electro-electric converter chip according to claim 2, characterized in that, The preset tilt angle is 0-45 degrees; or / and When performing electroporation on cells in the electroporation solution, both the first inner wall and the second inner wall are perpendicular to the horizontal plane.

4. The electro-electric converter chip according to claim 2, characterized in that, The ratio between the depth and width of the electro-rotation space ranges from 1:1 to 5:1; wherein, the depth of the electro-rotation space refers to the distance between the opening and a preset point on the bottom surface, and the preset point is any point on the bottom surface; the width of the electro-rotation space refers to the distance between the first inner sidewall and the second inner sidewall when both the first titanium electrode sheet and the second titanium electrode sheet are attached to the plastic support.

5. The electro-electric converter chip according to claim 1, characterized in that, The first surface is provided with a first connecting portion surrounding the through groove, and the first titanium electrode sheet is sealed and bonded to the first surface of the plastic support through the first connecting portion. The second surface is provided with a second connecting portion surrounding the through groove, and the second titanium electrode sheet is sealed and bonded to the second surface of the plastic support through the second connecting portion.

6. The electro-electric converter chip according to claim 5, characterized in that, The first connecting part is a first adhesive guide groove for guiding adhesive into the first surface and the first titanium electrode sheet; the second connecting part is a second adhesive guide groove for guiding adhesive into the second surface and the second titanium electrode sheet.

7. The electro-electric converter chip according to claim 1, characterized in that, The first titanium electrode sheet is provided with at least two first positioning holes; the plastic support is provided with at least two second positioning holes opposite to each of the first positioning holes; the second titanium electrode sheet is provided with at least two third positioning holes opposite to each of the second positioning holes; the electro-rotation chip further includes at least two first positioning posts, each of which is used to sequentially pass through the first positioning hole, the second positioning hole and the third positioning hole to position the first titanium electrode sheet, the plastic support and the second titanium electrode sheet.

8. The electro-electric converter chip according to claim 1, characterized in that, The first titanium electrode sheet has at least two fourth positioning holes; the second titanium electrode sheet has at least two fifth positioning holes; the plastic support member has second positioning posts and third positioning posts on opposite sides respectively; the second positioning posts are arranged opposite to the fourth positioning holes so as to position the first titanium electrode sheet and the plastic support member when each of the second positioning posts passes through the fourth positioning holes in a corresponding manner; the third positioning posts are arranged opposite to the fifth positioning holes so as to position the second titanium electrode sheet and the plastic support member when each of the third positioning posts passes through the fifth positioning holes in a corresponding manner.

9. The electro-electric converter chip according to claim 1, characterized in that, The first titanium electrode sheet includes a first titanium substrate and a first metal coating on the first titanium substrate; the second titanium electrode sheet includes a second titanium substrate and a second metal coating on the second titanium substrate.

10. The electro-electric converter chip according to claim 1, characterized in that, The plastic support is a modified part with low surface properties; and / or The outer surface of the plastic support is covered with a hydrophobic layer; or / and The surface roughness of the first titanium electrode sheet facing the first surface ranges from Ra1.0 to Ra3.0; the surface roughness of the second titanium electrode sheet facing the second surface ranges from Ra1.0 to Ra3.0.

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