Chip culture dish, chip culture dish preparation method, and array culture dish

Abstract

The application relates to a chip culture dish, a chip culture dish preparation method and an array culture dish in the field of biomedical technology. The chip culture dish comprises a transparent substrate, a transparent conductive film, the transparent conductive film is laid on one side of the transparent substrate, the transparent conductive film comprises a first electrode and a second electrode, a culture cavity is arranged on the side of the transparent substrate with the transparent conductive film, a culture cavity is formed in the culture cavity, the culture cavity covers the first electrode and the second electrode, a first conductive part is connected with the first electrode and used for leading out the first electrode, and a second conductive part is connected with the second electrode and used for leading out the second electrode. The chip culture dish can realize cell electric signal observation and image observation in the field of biomedicine at the same time, and the effect of microscopic observation is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biomedical technology, in particular to a chip culture dish, a chip culture dish preparation method and an array culture dish. BACKGROUND

[0002] The current cell electrofusion technology is developing rapidly, and needs to be integrated in multiple disciplines, including physical, chemical, electrical and other fields of research. In the field of cells, one challenge is to use a microscope with a high-power lens to observe the culture dish, and also to make the culture dish have the function of detecting electrical signals at the same time. Although the electrical impedance spectrum detection has been widely used in cell research, the materials in the cell electrical impedance spectrum detection chip generally use non-light-transmitting metal materials such as gold and silver. The use of non-light-transmitting metal materials will affect the microscopic observation effect of the cells, making the observation area in the well smaller, only 20% of the area is the observation area.

[0003] Therefore, the present application is proposed. SUMMARY

[0004] In view of the problems in the background art, the present application provides a chip culture dish, a chip culture dish preparation method and an array culture dish, which can simultaneously realize cell electrical signal observation and image observation in the field of biomedicine, and improve the microscopic observation effect.

[0005] According to a first aspect of the present application, a chip culture dish is provided, comprising: a transparent substrate, a transparent conductive film laid on one side of the transparent substrate, the transparent conductive film comprising a first electrode and a second electrode; a culture cavity provided on the side of the transparent substrate with the transparent conductive film, a culture cavity being formed in the culture cavity, the culture cavity covering the first electrode and the second electrode; a first conductive member connected with the first electrode for leading out the first electrode; and a second conductive member connected with the second electrode for leading out the second electrode.

[0006] In some embodiments of the present application, two or more through holes are formed on the transparent substrate, the through holes communicating the upper and lower sides of the transparent substrate; the two or more through holes are filled with conductive material, a part of which forms the first conductive member and another part of which forms the second conductive member.

[0007] In some embodiments of the present application, the transparent conductive film further comprises a first area connected with the first electrode and a second area connected with the second electrode, the first conductive member and the corresponding through hole are located in the first area, and the second conductive member and the corresponding through hole are located in the second area.

[0008] In some embodiments of the present application, the first area and the second area are both located outside the coverage of the culture cavity.

[0009] In some embodiments of the present application, the transparent conductive film is an ITO conductive film; the thickness of the ITO conductive film is 30-110 nm.

[0010] According to a second aspect of the present application, a chip culture dish preparation method is provided for preparing the chip culture dish described above, the chip culture dish preparation method comprising the following steps:

[0011] S1: preparing a transparent substrate, partitioning the transparent substrate according to the size of the chip culture dish to be prepared, and punching a set number of through holes in each partition of the transparent substrate;

[0012] S2: preparing a transparent conductive film corresponding to each partition on one side of the transparent substrate, the transparent conductive film comprising a first electrode and a second electrode;

[0013] S3: filling a conductive material in the through holes of each partition of the transparent substrate, respectively, a part of which forms a first conductive piece connected to the first electrode, and another part of which forms a second conductive piece connected to the second electrode;

[0014] S4: dicing the transparent substrate to separate the partitions to obtain a single transparent substrate;

[0015] S5: preparing a culture cavity on the side of the single transparent substrate with the transparent conductive film by a mold, so that the culture cavity covers the corresponding first electrode and second electrode, and finally obtaining a chip culture dish separated from each other.

[0016] In some embodiments of the present application, the conductive material in step S3 is a conductive paste, and the conductive paste is filled into the through holes by a vacuum hole filling machine.

[0017] Preferably, the conductive paste in the through holes is subjected to a vacuum operation and a pressure increasing operation to remove air bubbles, and the vacuum operation and the pressure increasing operation are repeated multiple times.

[0018] Preferably, the vacuum operation is performed until the air pressure is less than 5000 pa, and the pressure increasing operation is performed until the air pressure is one standard atmosphere.

[0019] In some embodiments of the present application, the transparent conductive film in step S2 is prepared by a magnetron sputtering process.

[0020] Preferably, before the transparent conductive film is prepared by sputtering, SiO2 is first plated on the transparent substrate.

[0021] In some embodiments of the present application, the through holes in step S1 are formed by a laser punching process.

[0022] According to a third aspect of the present application, there is provided an array culture dish, comprising: a plurality of the above-mentioned chip culture dishes, the plurality of chip culture dishes being arranged in an array; and a circuit board adapter, the circuit board adapter being arranged at the bottom of the plurality of chip culture dishes arranged in an array, the first conductive member and the second conductive member of each chip culture dish being connected to the circuit board adapter respectively.

[0023] The present application provides a chip culture dish, a chip culture dish preparation method and an array culture dish. The present application uses laser drilling process, magnetron sputtering process, vacuum hole plugging process, cutting process and vacuum replication process to realize batch production of the chip culture dish based on the structure of transparent substrate, transparent conductive film, microelectrode array and culture cavity. The chip culture dish can be expanded to 12-well, 24-well, 48-well and 96-well array culture dishes by using a circuit board adapter. The chip culture dish can realize comprehensive observation of cells in the culture cavity by using a confocal device and real-time detection and analysis of the electrical impedance spectrum of cells by using an electrical signal detection device, so as to realize cell electrical signal observation and high-quality image observation in the biomedical field at the same time, and allow multiple electrodes to be placed at the same time instead of being placed separately. The chip culture dish can set up a control in the same experimental setup (using one electrode as a control and other electrodes as experiments), is suitable for culturing cells and microscope observation, and can be well applied to drug safety evaluation and drug high-throughput screening. BRIEF DESCRIPTION OF DRAWINGS

[0024] Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The accompanying drawings are included to provide a description of the preferred embodiments and are not intended to limit the scope of the application. Furthermore, the same reference numerals are used throughout the several views of the drawings to denote the same or similar parts. In the drawings:

[0025] Figure 1 is a front view of the chip culture dish of the present application;

[0026] Figure 2 is a front view of the chip culture dish of the present application; Figure 1 is a sectional view in the direction of A-A in

[0027] Figure 3 is a top view of the chip culture dish of the present application;

[0028] Figure 4 is a schematic view of the transparent substrate and the structure thereon of the present application;

[0029] Figure 5 is a preparation process diagram of the chip culture dish of the present application;

[0030] Figure 6 is a schematic view of an array culture dish of the present application;

[0031] Figure 7 is a picture of the transparent substrate prepared in Example 1;

[0032] Figure 8 is a picture of the finished chip culture dish prepared in Example 1.

[0033] In the drawings: 1, transparent substrate; 11, through hole; 2, transparent conductive film; 21, first electrode; 22, second electrode; 23, first piece area; 24, second piece area; 3, culture cavity; 31, culture cavity; 4, first conductive piece; 5, second conductive piece; 6, well plate; 7, transparent base; 8, first circuit board; 9, second circuit board; 10, plug-in connection structure. DETAILED DESCRIPTION

[0034] It should be clear that the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application.

[0035] The following description refers to the accompanying drawings. In the drawings, like reference numbers indicate identical or similar elements, unless otherwise indicated. The following description of exemplary embodiments is not meant to represent all embodiments in accord with the present application. Rather, they are presented as examples of devices and methods consistent with some aspects of the present application as detailed in the appended claims.

[0036] In the description of the present application, it should be understood that the terms "first", "second", etc. are used only for the purpose of description, and cannot be understood as indicating or implying relative importance. For those of ordinary skill in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances. In addition, in the description of the present application, "multiple" means two or more, unless otherwise specified. "And / or", which describes the relationship between the associated objects, means that there can be three relationships, for example, A and / or B can mean that there are three cases of A alone, A and B together, and B alone. The character " / " generally represents an "or" relationship between the associated objects.

[0037] The present application discloses a chip culture dish, as shown in Figures 1-4 The chip culture dish includes a transparent substrate 1, a transparent conductive film 2, a culture cavity 3, a first conductive piece 4, and a second conductive piece 5.

[0038] The transparent conductive film 2 is laid on one side of the transparent substrate 1, and the transparent conductive film 2 includes a first electrode 21 and a second electrode 22.

[0039] The culture cavity 3 is arranged on the side of the transparent substrate 1 with the transparent conductive film 2, and a culture cavity 31 is formed in the culture cavity 3, and the culture cavity 31 covers the first electrode 21 and the second electrode 22.

[0040] The first conductive member 4 and the second conductive member 5 are connected with the first electrode 21 and the second electrode 22 respectively, the first conductive member 4 is used to lead out the first electrode 21, and the second conductive member 5 is used to lead out the second electrode 22.

[0041] By using the chip culture dish in the technical solution, the culture medium containing cells is added in the culture cavity 31 of the culture cavity 3, and then the chip culture dish is placed in a cell culture box for culture, when it is needed to observe and test the cells, the chip culture dish can be taken out from the cell culture box, the chip culture dish is connected with the electric signal detection equipment through the first conductive member 4 and the second conductive member 5, and the light transmission and conductivity of the transparent conductive film 2 are utilized, so that the cells in the culture cavity 31 can be comprehensively observed by using a confocal equipment (a microscope with a high-power lens), and the electric impedance spectrum of the cells can be detected and analyzed in real time by using the electric signal detection equipment through the first electrode 21 and the second electrode 22, so that the cell electric signal observation and high-quality image observation in the biomedical field are realized at the same time, and the effect of microscopic observation is improved.

[0042] In some embodiments of the present application, as shown in Figure 2 The culture cavity 3 can adopt a hollow tubular structure, the culture cavity 31 is formed in the internal hollow of the hollow tubular structure, and the culture cavity 31 takes the surface of the transparent substrate 1 as a bottom surface, and the top opening of the culture cavity 31 is used for adding, replacing and removing the culture medium.

[0043] It should be noted that the first electrode 21 and the second electrode 22 are located in the internal hollow range of the hollow tubular structure, that is, the size of the culture cavity 31, such as the diameter, can be designed according to the distribution range of the first electrode 21 and the second electrode 22 on the transparent substrate 1, so that the culture cavity 31 is matched with the first electrode 21 and the second electrode 22.

[0044] Further, the solid part of the hollow tubular structure can partially or entirely cover other areas of the transparent substrate 1 (the transparent conductive film 2), preferably entirely cover, that is, the outer contour of the culture cavity 3 close to the side of the transparent substrate 1 is equivalent to or slightly larger than the outer contour of the transparent substrate 1.

[0045] In addition, the outer contour of the culture cavity 3 away from the side of the transparent substrate 1 can be equivalent to or slightly smaller than the outer contour of the transparent substrate 1, preferably smaller; and the shape of the outer contour of the culture cavity 3 away from the side of the transparent substrate 1 can be the same as or different from the outer contour of the transparent substrate 1.

[0046] For example, in the present application, the transparent substrate 1 is rectangular, the culture cavity 3 is designed as a rectangular near the side of the transparent substrate 1, and the outer contour of the culture cavity 3 far from the side of the transparent substrate 1 is designed as a circle, that is, a combination of a circular tube cavity structure and a square tube cavity structure.

[0047] In some embodiments of the present application, the material of the culture cavity 3 includes, but is not limited to, medical grade PS (polystyrene) and the like, which has the advantages of non-toxicity and high safety, and is beneficial to cell culture.

[0048] In some embodiments of the present application, as shown in Figure 4 Two or more through holes 11 are formed on the transparent substrate 1, the through holes 11 communicate the upper and lower sides of the transparent substrate 1, and the through holes 11 are filled with conductive materials, part of which forms the first conductive part 4 and part of which forms the second conductive part 5.

[0049] In the present application, the side of the transparent substrate 1 with the transparent conductive film 2 is taken as the top surface, and the side without the transparent conductive film 2 is taken as the bottom surface. The conductive materials in the through holes 11 are used to connect the bottom surface and the top surface, and in the case of direct or indirect connection between the conductive materials and the first electrode 21 and the second electrode 22, the electrode electrical signal is led out from the bottom surface of the transparent substrate 1. On the one hand, the stability and damage resistance are higher, and on the other hand, the chip culture dish can be conveniently connected with the electrical signal detection equipment, improving the operation convenience and connection effect.

[0050] Further, the conductive material can be selected from conductive paste, such as silver paste, copper paste, solder, conductive resin and the like.

[0051] Further, the conductive paste can be filled into the through holes 11 by a vacuum hole plugging machine, and after curing, the first conductive part 4 and the second conductive part 5 are formed respectively, so that the transparent conductive film 2 can be used for signal measurement and led out from the back surface by the hole plugging process, realizing signal measurement on one side of the transparent substrate 1 and signal leading out on the other side.

[0052] In other embodiments of the present application, the first conductive part 4 and the second conductive part 5 can also be selected to be arranged on the side wall around the transparent substrate 1, and further extended to the back surface of the transparent substrate 1, so as to lead out the first electrode 21 and the second electrode 22 from the periphery of the transparent substrate 1 or the back surface of the transparent substrate 1 respectively.

[0053] Specifically, silver paste can be used on the side wall around the transparent substrate 1 to form the first conductive part 4 and the second conductive part 5 at the side wall position.

[0054] In some embodiments of the present application, as shown in Figure 2 and Figure 4As shown, the transparent substrate 1 is formed with a contact of the hole disc 6 at each through hole 11 on the bottom surface thereof for transmitting the electrode electric signal, i.e. further improving the connection stability and convenience of the chip culture dish and the electric signal detection device.

[0055] In some embodiments of the present application, as shown in Figure 4 As shown, the transparent conductive film 2 further comprises a first piece area 23 connected with the first electrode 21 and a second piece area 24 connected with the second electrode 22, and it should be understood that the first conductive member 4 and the corresponding through hole 11 are located in the first piece area 23, and the second conductive member 5 and the corresponding through hole 11 are located in the second piece area 24.

[0056] By connecting the first electrode 21 with the first conductive member 4 through the first piece area 23 and connecting the second electrode 22 with the second conductive member 5 through the second piece area 24, the first electrode 21 and the second electrode 22 can be better led out.

[0057] In some embodiments of the present application, as shown in Figure 2 and Figure 4 As shown, the first piece area 23 and the second piece area 24 are both located in the area outside the coverage range of the culture cavity 31.

[0058] It should be understood that the first piece area 23 and the second piece area 24 are located in the area covered by the solid part of the culture cavity 3, the first piece area 23 is an integral structure with the first electrode 21, and the second piece area 24 is an integral structure with the second electrode 22.

[0059] By setting the first piece area 23 and the second piece area 24 in the solid part of the culture cavity 3, the effect of the culture cavity 31 area can be avoided, and the test effect can be ensured.

[0060] In some embodiments of the present application, as shown in Figure 4 As shown, the number of through holes 11 on the first piece area 23 and the second piece area 24 respectively includes but is not limited to 1, 2, 3 or 4, etc., and the through holes 11 on the first piece area 23 and the second piece area 24 are arranged in a spaced or uniform distribution manner.

[0061] In some embodiments of the present application, as shown in Figure 4 As shown, the first electrode 21 and the second electrode 22 can adopt an interdigital electrode, i.e. the first electrode 21 and the second electrode 22 are respectively a comb-shaped (finger-shaped) microelectrode array (MEA), and it should be understood that the microelectrode array and the transparent substrate 1 together form a microelectrode chip.

[0062] It should be noted that the first area 23 and the second area 24 can be regular or irregular planar patterns, the first area 23 and the second area 24 are spaced apart by a certain distance, each line of the first electrode 21 is connected to the side of the first area 23 close to the second area 24, and each line of the second electrode 22 is connected to the side of the second area 24 close to the first area 23, thereby forming the interdigital electrode.

[0063] Further, in the preparation of the transparent conductive film 2, the comb-shaped in-plane periodic pattern electrode can be formed on the transparent substrate 1 by photolithography, magnetron sputtering and stripping process to obtain a super-fine circuit as a core component for electrical signal transmission, and at the same time, the first area 23 and the second area 24 are obtained.

[0064] In other embodiments of the present application, the transparent conductive film 2 can also be prepared on the transparent substrate 1 by evaporation.

[0065] Further, the line-to-line spacing of the interdigital electrode (the spacing between adjacent lines of the first electrode 21 and the second electrode 22) can be 40-120 μm, and the line width of the interdigital electrode (the line width of the first electrode 21 and the second electrode 22) can be 40-120 μm.

[0066] It should be noted that the line-to-line spacing of the interdigital electrode and the line width of the interdigital electrode can be reasonably designed according to the needs of the person skilled in the art, for example, the line-to-line spacing of the interdigital electrode can be 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120 μm, etc., and the line width of the interdigital electrode can be 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120 μm, etc.

[0067] In some embodiments of the present application, the transparent conductive film 2 can be an ITO conductive film.

[0068] Further, the thickness of the ITO conductive film can be 30-110 nm, i.e., the thickness of the first electrode 21 and the second electrode 22 and the first area 23 and the second area 24 is 30-110 nm.

[0069] The ITO conductive film has good biocompatibility, conductivity, and high light transmittance in the visible light band, and the thickness design has less interference with medical imaging equipment, which is beneficial to the observation of the microscope and the electrical impedance spectroscopy detection analysis of the electrical signal detection equipment, thereby realizing cell electrical signal detection while effectively expanding the area of microscope image observation.

[0070] It should be noted that the thickness of the ITO conductive film can be reasonably designed by those skilled in the art as needed, and for example, the thickness of the ITO conductive film can be 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 110 nm, etc.

[0071] In other embodiments of the present application, the transparent conductive film 2 can also use other transparent conductive metal oxide films, and those skilled in the art can reasonably select the material of the transparent conductive film 2 and design the thickness of various transparent conductive films 2 as needed, preferably using an ITO conductive film.

[0072] In some embodiments of the present application, as shown in Figure 4 The transparent substrate 1 can be square, rectangular or circular in shape, and preferably square.

[0073] Further, the material of the transparent substrate 1 includes but is not limited to glass, transparent plastic, etc.

[0074] In some embodiments of the present application, the length and width of the transparent substrate 1 can be independently 10-14 mm, and the thickness of the transparent substrate 1 can be 0.4-1.3 mm. Those skilled in the art can reasonably design the size of the microelectrode chip composed of the transparent substrate 1 and the electrode as needed.

[0075] In some embodiments of the present application, as shown in Figure 2 The first embedding groove is formed on the end face of the culture cavity 3 close to one end of the transparent substrate 1, and the second embedding groove is formed at the bottom of the first embedding groove. The edges of the second embedding groove are spaced apart from the edges of the first embedding groove, so that the first embedding groove and the second embedding groove form a stepped groove structure, and the transparent substrate 1 is accommodated in the second embedding groove.

[0076] In this embodiment, by placing the transparent substrate 1 in the inner embedding groove of the stepped groove, the transparent substrate 1 can be protected, effectively avoiding damage to the transparent substrate 1 and the hole disc 6 structure, and facilitating the connection of the hole disc 6 and the external lead.

[0077] Preferably, after the transparent substrate 1 is accommodated in the second embedding groove, the surface of the transparent substrate 1 does not protrude from the bottom of the first embedding groove.

[0078] Preferably, the hole disc 6 does not protrude from the corresponding end face of the culture cavity 3.

[0079] Further, the hole disc 6 is spaced apart from the corresponding end face of the culture cavity 3.

[0080] The present application also provides a chip culture dish preparation method for preparing the above chip culture dish, as shown in Figure 5As shown, the chip culture dish preparation method comprises the following steps:

[0081] S1: preparing a transparent substrate, partitioning the transparent substrate according to the size of the prepared chip culture dish, and punching a set number of through holes in each partition of the transparent substrate.

[0082] S2: preparing a transparent conductive film corresponding to each partition on one side of the transparent substrate, the transparent conductive film comprising a first electrode and a second electrode.

[0083] S3: filling the through holes in each partition of the transparent substrate with conductive material, respectively, a part of which forms a first conductive part connected to the first electrode, and another part forms a second conductive part connected to the second electrode.

[0084] S4: dicing the transparent substrate to divide each partition to obtain a single transparent substrate.

[0085] S5: preparing a culture cavity on the side of the single transparent substrate with the transparent conductive film through a mold, so that the culture cavity of the culture cavity covers the corresponding first electrode and second electrode, and finally obtains a chip culture dish separated from each other.

[0086] By using the preparation method of the application, the rapid mass production of ex vivo electrophysiological microelectronic array (MEA) chip culture dishes can be realized, and the uniform and repeatable preparation of high-throughput chips is also realized. The application has great potential in batch production, has the advantages of relatively low production cost and high production efficiency, and provides a complete preparation method for the industrialization and batch production of MEA type culture dishes.

[0087] In some embodiments of the application, the conductive material in step S3 is conductive paste, and the conductive paste is filled into the through holes by a vacuum hole filling machine.

[0088] The application can avoid the problem of surface depression caused by insufficient filling by filling the conductive paste into the through holes by a vacuum hole filling machine, which is beneficial to reducing the fine line production and the numerical value of the characteristic impedance of the conductive paste after curing.

[0089] Preferably, the conductive paste in the through holes is removed by vacuum operation and pressure increasing operation to remove bubbles, and the vacuum operation and pressure increasing operation are repeated for 3-9 times.

[0090] Preferably, the vacuum operation is performed until the air pressure is less than 5000 pa, and the pressure increasing operation is performed until the air pressure is one standard atmosphere.

[0091] In some embodiments of the application, the through holes in step S1 are formed by a laser punching process.

[0092] The application forms the through hole by the laser drilling process, has the advantages of high drilling speed, high efficiency and good economic benefit, and the formed through hole has high quality, micron-level processing of the through hole can be realized, and the damage to the transparent substrate is reduced.

[0093] In some embodiments of the application, the transparent conductive film in step S2 is prepared by a magnetron sputtering process. The basic principle of the magnetron sputtering method is that under the action of an electric field and an alternating magnetic field, high-energy particles accelerated by the electric field bombard the surface of the target material, atoms on the surface of the target material are separated from the original lattice and escape, and then transferred to the surface of the transparent substrate to form a transparent conductive film.

[0094] Preferably, before the transparent conductive film is prepared by sputtering, SiO2 is first plated on the transparent substrate.

[0095] The application uses a direct current magnetron sputtering method to continuously plate a transparent conductive film, which has the advantages of uniform film thickness, easy control, good film repeatability, stability, large area plating, arbitrary placement of the relative position of the transparent substrate and the electrode pattern according to the layout, preparation of a dense film layer at low temperature, alloy target reaction sputtering, direct sputtering of an oxidation target, and many other advantages.

[0096] In some embodiments of the application, the culture cavity can be prepared by a vacuum replication process or an injection molding process in step S5.

[0097] The application also provides an array-type culture dish, which comprises a plurality of the above-mentioned chip culture dishes and a circuit board adapter.

[0098] The plurality of chip culture dishes are arranged in an array type, the circuit board adapter is arranged at the bottom of the plurality of chip culture dishes arranged in an array type, and the first conductive member and the second conductive member of each chip culture dish are connected with the circuit board adapter, respectively.

[0099] Through the circuit board adapter, the array-type culture dish for ex vivo electrophysiological microelectrodes with the number of 12 holes, 24 holes, 48 holes and 96 holes, etc. can be expanded, which allows multiple electrodes to be placed simultaneously instead of being placed individually, can perform multiple parameter optimization experiments in the same experimental setup, uses one electrode (culture cavity) as a control and other electrodes (culture cavities) as experiments, is suitable for culturing cells and microscope observation, can be well applied to drug safety evaluation and high-throughput drug screening, and the experimenter can define the number of groups needed, which can save the experimental cost of the user to a certain extent and reduce waste.

[0100] In some embodiments of the application, the array-type arrangement of the plurality of chip culture dishes can be achieved by split splicing or by one-piece molding.

[0101] In one embodiment of the application, as shown in Figure 6As shown, the plurality of chip culture dishes are integrally formed, and the circuit board adapter comprises a transparent base 7 fixed thereunder, the transparent base 7 is provided with a first circuit board 8 at one end and a second circuit board 9 at the other end, the first conductive member 4 of each culture cavity 31 is independently led out to and connected with the first circuit board 8, and the second conductive member 5 of each culture cavity 31 is independently led out to and connected with the second circuit board 9.

[0102] In some embodiments of the present application, the first circuit board 8 and the second circuit board 9 can adopt a flexible circuit board (FPC) or a rigid circuit board (PCB), and preferably adopt a flexible circuit board.

[0103] Further, the other end of the first circuit board 8 or the second circuit board 9 can be provided with a plug-in connection structure 10 for connecting with an external detection device.

[0104] It should be understood that the first conductive member 4 of each culture cavity 31 is connected with the corresponding plug-in connection structure 10 through the printed conductive circuit in the first circuit board 8, and the second conductive member 5 of each culture cavity 31 is connected with the corresponding plug-in connection structure 10 through the printed conductive circuit in the second circuit board 9.

[0105] Further, in some embodiments of the present application, as shown, Figure 6 The plug-in connection structure 10 comprises a pin array, an FPC plug-in connector, a flat plate type rigid circuit board and a male (female) connector, wherein the pin array is connected with the flexible circuit board, the FPC plug-in connector is arranged on one side of the flat plate type rigid circuit board and is adapted with the pin array, the male (female) connector is arranged on the other side of the flat plate type rigid circuit board and can be electrically connected with the detection device through the female (male) connector, that is, the flat plate type rigid circuit board, the FPC plug-in connector and the male (female) connector thereon serve as an intermediate structure to detachably connect the first circuit board 8 (the second circuit board 9) with the detection device.

[0106] The chip culture dish preparation method in the present application will be further described below in conjunction with specific examples.

[0107] Example 1

[0108] Step 1: The process flow of the punching process is: feeding → CCD visual positioning → laser punching → discharging → rotation / broken hole detection.

[0109] After the glass is cleaned and dried in the previous process, it is conveyed to the processing table, the glass is accelerated and separated, the glass spacing is enlarged, and time interval is left for subsequent punching. After accelerated conveying, the glass enters the feeding conveying table.

[0110] Each cleaned glass substrate, size is 200 mm*200 mm, thickness is 0.7 mm, after punching a hole in the glass substrate by laser, the aperture size is 0.6 mm, using etching solution to remove the edge collapse, cleaning, drying or air drying the solution.

[0111] The punching position of the glass on the laser workstation is captured by a CCD vision system, and the product position and attitude are judged to complete accurate positioning.

[0112] Then use etching solution to remove the edge collapse, clean, dry or air dry the solution.

[0113] Step 2: Use a magnetron sputtering process to sputter ITO target material according to the design pattern.

[0114] First, prepare 0.7mm ultra-thin glass sheets of a specified thickness and size, wash them with deionized water, and clean them with ultrasonic waves. After entering the vacuum chamber, first coat SiO2, then enter the ITO coating chamber to coat ITO film. After heating, curing and annealing, the finished product is obtained.

[0115] The ITO conductive film glass is made by depositing indium tin oxide film on the surface of the glass sheet through magnetron sputtering and heating annealing under high vacuum and dust-free conditions with Ar (argon) and O2 (oxygen) gas.

[0116] A photoresist layer is coated on the glass, baked, and then exposed and developed according to the design layout. Then the sample is cleaned using oxygen plasma, ITO is deposited on the surface of the sample by magnetron sputtering, and then acetone is used for stripping to form regular microelectrodes, a total of 255, and then cleaned and dried.

[0117] The microelectrode is an interdigital electrode, with a size of 12 mm*12 mm, a pitch of 80 μm, a line width of 100 μm, and a thickness of 50 nm.

[0118] Step 3: Use a vacuum hole plugging machine to fill the through hole with paste.

[0119] The process flow of this step is: vacuum pumping - loading screen plate and pad - alignment - equipment vacuum pumping - trial printing - inspection.

[0120] To make the screen plate on the ITO-free side, make the silk screen plate according to the drawing, and use screen printing process to fill the hole disc. The steel mesh needs to protect the electrode part from being contaminated by the hole plugging material. The hole plugging material is coated on the surface of the silk screen plate, and the hole plugging material is in a liquid state, so that the hole plugging material flows naturally under the action of gravity.

[0121] The hole plugging material is selected from copper paste.

[0122] The vacuumizing operation is performed to make the air inside the plug material leave the plug material and form bubbles on the surface of the plug material, and the pressure boosting operation is performed to break the bubbles remaining on the surface of the plug material; the glass is placed into a vacuum drying box to perform the vacuumizing operation, and the air pressure in the vacuum drying box should be less than 5000 Pa, preferably 100 Pa, 500 Pa or 1000 Pa, and specifically 500 Pa.

[0123] At this time, the plug material is observed, when bubbles are generated on the surface of the plug material and part of the bubbles are broken, the vacuumizing operation is stopped, and the external air is allowed to enter, at this time, the external air flows in a large amount, the air pressure rises to one standard atmosphere, and the remaining part of the bubbles is broken by the pressure.

[0124] The vacuumizing operation and the pressure boosting operation are repeated until no bubbles visible to the naked eye are generated; the vacuumizing operation and the pressure boosting operation are repeated 3-9 times, preferably 5 times, until no bubbles visible to the naked eye are generated.

[0125] Then, the ethanol is cleaned.

[0126] Step 4: The ITO glass is cut by using a laser or a hob, and is cut into a single piece with a size of 12 mm*12 mm.

[0127] In order to further improve the processing precision and quality, the laser wavelength 532 nm is used, the repetition frequency coverage range is wide (single pulse to 200 kHz), the beam quality (M<2) is superior, and is strictly guaranteed in all frequency ranges; the pulse width is <25 ns, that is, the nanosecond green laser is used for micro-nano processing.

[0128] When the nanosecond green laser is processed, the heat affected zone is very small, the processing speed is fast, the heat affected zone is small, the focusable spot size is small, and the like, and high processing precision and quality are easily obtained during micro-processing.

[0129] The process parameters include: energy 30±0.1 W, frequency 150 KHz, cutting speed 400±20 mm / s, and spiral turns 4000±2 turns.

[0130] After the parameters are determined, a laser cutting machine is used to cut according to the design drawing, and a single piece is formed, as shown in FIG. 5. Figure 7

[0131] After the cutting is completed, alcohol is used for cleaning, and drying is performed.

[0132] Step 5: The single piece of glass is placed into a culture dish mold, a medical grade polystyrene solution is used to perform a vacuum laminating process, after 3 hours, dry ice is used for cleaning, and a chip culture dish is obtained, as shown in FIG. 6. Figure 8

[0133] ​​The above merely describes preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, any changes or replacements within the technical scope disclosed by the present application, which can be easily thought by those skilled in the art, should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A chip culture dish, characterized by, The chip culture dish comprises: a transparent substrate; a transparent conductive film laid on one side of the transparent substrate, the transparent conductive film comprising a first electrode and a second electrode; the transparent conductive film further comprising a first area connected with the first electrode and a second area connected with the second electrode; a culture cavity provided on the side of the transparent substrate with the transparent conductive film, the culture cavity being formed in the culture cavity body, and the culture cavity covering the first electrode and the second electrode; a first conductive part connected with the first electrode for leading out the first electrode; a second conductive part connected with the second electrode for leading out the second electrode; two or more through holes are formed on the transparent substrate, the through holes being communicated between the upper and lower sides of the transparent substrate, the two or more through holes being filled with conductive material, a part of which forms the first conductive part and another part of which forms the second conductive part, the first conductive part and the corresponding through hole being located in the first area, and the second conductive part and the corresponding through hole being located in the second area; a contact of a hole disc is formed on the bottom surface of the transparent substrate at each through hole, a ring-shaped first embedding groove is formed on the end surface of the culture cavity body close to the transparent substrate, and a second embedding groove is formed at the bottom of the first embedding groove, the edge of the second embedding groove being spaced apart from the edge of the first embedding groove to form a stepped groove structure, the transparent substrate is accommodated in the second embedding groove, and the hole disc does not protrude from the corresponding end surface of the culture cavity body.

2. The chip-plate according to claim 1, wherein, The first area and the second area are located in the area outside the coverage range of the culture cavity.

3. The chip-plate according to claim 1 or 2, characterized in that The transparent conductive film is an ITO conductive film. The thickness of the ITO conductive film is 30-110 nm.

4. A method for preparing a chip dish for preparing a chip dish according to any one of claims 1 to 3, characterized in that The chip culture dish preparation method comprises the following steps: S1: preparing a transparent substrate, partitioning the transparent substrate according to the size of the prepared chip culture dish, and punching a set number of through holes in each area of the transparent substrate; S2: preparing a transparent conductive film corresponding to each area on one side of the transparent substrate, the transparent conductive film comprising a first electrode and a second electrode; S3: filling conductive material in the through holes of each area of the transparent substrate, a part of which forming a first conductive part connected with the first electrode and another part of which forming a second conductive part connected with the second electrode; S4: scribing the transparent substrate to divide the areas to obtain a single transparent substrate; S5: preparing a culture cavity body on one side of the single transparent substrate with the transparent conductive film by a mold, so that the culture cavity of the culture cavity body covers the corresponding first electrode and second electrode, and finally a mutually separated chip culture dish is obtained.

5. The chip-plate preparation method of claim 4, wherein, The conductive material in step S3 is conductive paste, and the conductive paste is filled into the through holes by a vacuum hole filling machine; The conductive paste in the through holes is subjected to a vacuum operation and a pressure increasing operation to remove air bubbles, and the vacuum operation and the pressure increasing operation are repeated multiple times; The vacuum operation is performed until the air pressure is less than 5000 pa, and the pressure increasing operation is performed until the air pressure reaches one standard atmosphere.

6. The chip-plate preparation method of claim 4, wherein, The transparent conductive film in step S2 is prepared by a magnetron sputtering process; Before the transparent conductive film is prepared by sputtering, SiO2 is first plated on the transparent substrate.

7. The chip-plate preparation method of claim 4, wherein, The through hole in the step S1 is formed by a laser drilling process.

8. An arrayed culture dish, characterized by, The application relates to a chip culture dish. A plurality of chip culture dishes as claimed in any one of claims 1-3 are arranged in an array. A circuit board adapter is arranged at the bottom of the array of chip culture dishes, and the first conductive member and the second conductive member of each chip culture dish are connected to the circuit board adapter, respectively.

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