Transwell-based electrophysiological information acquisition interface device

Abstract

This invention discloses a transwell-based electrophysiological information acquisition interface device, comprising a cell porous culture medium holder, a PCB signal transmission module, and several signal acquisition units. The cell porous culture medium holder includes a base body, transwell insertion holes, and perfusion channels, with each perfusion channel connected to at least one transwell insertion hole. The PCB signal transmission module includes a PCB board, adapter components, and gold finger electrodes. The PCB board has several mounting holes, and the adapter components are respectively disposed on the outer periphery of each mounting hole and electrically connected to the corresponding gold finger electrodes. Each signal acquisition unit is equipped with a transwell and a flexible microelectrode array. One end of the flexible microelectrode array is electrically connected to the adapter components on the outer periphery of the mounting holes, and the other end is placed inside the transwell. This invention can simultaneously acquire and analyze the electrophysiological information of cells cultured in different transwells and achieve information exchange between cells in different transwells through a common cell culture medium perfusion channel.

Description

Technical Field

[0001] This invention belongs to the field of cell electrophysiological information acquisition technology in biomedical engineering, and specifically relates to an electrophysiological information acquisition interface device based on transwell. Background Technology

[0002] Traditional electrophysiological data acquisition interfaces include patch-clamp devices or microelectrode arrays (MEAs). The patch-clamp technique involves placing a microglass tube electrode (patch electrode or patch pipette) into the cell membrane, sealing it with an impedance of gigahertz or higher. This electrically isolates a small region of the cell membrane (the patch) at the electrode tip opening from its surroundings, allowing for the fixation of a point and the monitoring and recording of ion currents in the ion channels on the patch. While the patch-clamp technique offers high resolution for recording cell membrane channel currents, it can only operate on a single channel, resulting in low throughput, and requires a high level of operator skill.

[0003] MEA plates are sensors that use microelectromechanical systems (MEMS) to deposit metal onto a glass plate, forming a passivation layer, electrodes, and leads. These plates are used to transmit and detect changes in the action potentials of individual cells and the transmission of information between multi-cell networks. MEAs offer advantages such as a high number of detection channels, non-destructive testing, and long-term monitoring capabilities. However, MEA plates can only detect the action potentials or local field potentials of isolated cells in a single environment and cannot simultaneously culture multiple cell types for detection.

[0004] Transwells are commercially available standard devices for culturing cells and conducting cell experiments. Also known as cell chambers, they are small cups that can be placed inside well plates. The bottom of each cup is a polycarbonate JIE semi-permeable membrane with pore sizes ranging from 0.1 to 12 μm. Transwells can be used for co-culture, drug delivery, tissue reconstruction, and other cell experiments. While transwells can perform various cell experiments, cells placed in different transwells within a standard well plate cannot exchange signals or have their electrical signals acquired, making it impossible to analyze the electrophysiological information generated during cell culture. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides a transwell-based electrophysiological information acquisition interface device that can simultaneously acquire and analyze the electrophysiological information of cells cultured in different transwells, and facilitate information exchange between cells in different transwells through a common cell culture medium perfusion channel.

[0006] The technical solution of the present invention is as follows:

[0007] An electrophysiological information acquisition interface device based on transwell includes a cell porous culture medium base, a PCB signal transmission module, and several signal acquisition units. The cell porous culture medium base includes a base body and several transwell insertion holes and at least one perfusion channel arranged within the base body. The perfusion channel is connected to at least one transwell insertion hole. The PCB signal transmission module includes a PCB board, several adapter components, and several gold finger electrodes arranged on the PCB board. The PCB board has mounting holes for use with the transwell insertion holes. The adapter components are respectively disposed on the outer periphery of each mounting hole and electrically connected to the corresponding gold finger electrodes. The signal acquisition units are configured with transwells and flexible microelectrode arrays. The transwells can be inserted into the mounting holes and placed within the transwell insertion holes. One end of the flexible microelectrode array is electrically connected to the adapter components on the outer periphery of the mounting holes, and the other end is placed within the transwell for acquiring electrophysiological information of cells cultured within the transwell.

[0008] As an alternative, the infusion channel has at least two channels.

[0009] As an optional solution, the transwell insertion holes are arranged in a rectangular or circular array; the shape and size of the transwell insertion holes and mounting holes are adapted to the transwell.

[0010] As an alternative, the cell porous culture medium seat is made of a biocompatible transparent material.

[0011] As an alternative, the PCB signal transmission module is fixed to the base body of the cell porous culture medium using nano double-sided adhesive.

[0012] As an optional solution, the transwell in the signal acquisition unit has a cup body, a neck, and a retaining ring connected sequentially from bottom to top; the bottom of the cup body is a semi-permeable membrane, through which the exchange of substances between the cup body and the infusion channel is realized; the retaining ring is used in conjunction with the mounting hole to fix the transwell in the corresponding transwell insertion hole.

[0013] As an optional solution, the electrophysiological information acquisition interface device also includes a cover for use with a cell porous culture medium stand.

[0014] As an optional solution, the adapter assembly includes pad insertion holes disposed around the mounting holes, and matching dual-row through-hole females and dual-row through-hole pins; the dual-row through-hole females are inserted into the pad insertion holes and electrically connected to the corresponding gold finger electrodes through the pad insertion holes; the lower pins of the dual-row through-hole pins are inserted into the dual-row through-hole females, and the upper pins are electrically connected to one end of the flexible microelectrode array.

[0015] As an optional solution, the signal acquisition unit is also equipped with a connector; the transwell, flexible microelectrode array, and dual-row through-hole pin header are fixedly connected through the connector to form a whole.

[0016] As an alternative, the connector has a socket and a slot; the lower pins of the double-row straight pin header pass through the socket and are inserted into the double-row straight pin header nut; the connector is engaged with the transwell through the slot.

[0017] The transwell-based electrophysiological information acquisition interface device disclosed in this invention has the following beneficial effects:

[0018] (1) Compared with patch clamp technology, the electrophysiological information acquisition interface device of the present invention is equipped with multiple transwells, each transwell being an independent signal acquisition channel, which can realize the simultaneous acquisition of electrophysiological information of cells cultured in the transwell through multiple channels; compared with traditional MEA plates, the electrophysiological information acquisition interface device can realize the acquisition of cell action potentials and local field potentials through multiple channels, and can also culture multiple types of cells and acquire electrophysiological information at the same time.

[0019] (2) This invention can not only realize the information exchange between cells in different transwells through the common cell culture medium perfusion channel of multiple interconnected transwell plug holes, and construct systems such as organoid culture and multi-organ interaction, but also conduct comparative experiments on different cell culture medium perfusion channels.

[0020] (3) The present invention can also design the signal acquisition unit as a pluggable structure by means of an adapter component composed of double row straight plug female and double row straight plug male, so as to realize the quick and flexible replacement of the signal acquisition unit.

[0021] (4) The present invention can also integrate the transwell, flexible microelectrode array, and dual-row through-hole pins into a single unit using connectors configured in the signal acquisition unit, facilitating flexible replacement of the signal acquisition unit. Furthermore, the present invention can also combine the connectors with the use of a robotic arm to achieve automated replacement of the transwell. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the overall structure of the transwell-based electrophysiological information acquisition interface device.

[0023] Figure 2 This is a schematic diagram of the layered structure of a transwell-based electrophysiological information acquisition interface device.

[0024] Figure 3a This is a schematic diagram of the structure of a porous cell culture medium seat; Figure 3b This is a cross-sectional view of a porous cell culture medium stand.

[0025] Figure 4 This is a partial structural diagram of a PCB signal transmission module;

[0026] Figure 5 This is an exploded view of the transwell signal acquisition unit.

[0027] Figure captions: 1-Porous cell culture medium holder, 11-Transwell insertion hole, 12-Cell culture medium perfusion channel, 13-Base body; 2-PCB signal transmission module, 21-PCB board, 22-Dual row straight-through female connector, 23-Dual row straight-through male connector, 24-Gold finger electrode, 25-Mounting hole, 26-Pad insertion hole; 3-Transwell signal acquisition unit, 31-Transwell, 311-Cup body, 312-Neck, 313 Snap ring, 32-Snap fastener, 321-Insert port, 322-Slot, 33-Flexible microelectrode array, 331-Connection hole, 332-Electrode array point; 4-Top cover. Detailed Implementation

[0028] The technical solution of the present invention will now be clearly and completely described with reference to specific embodiments and accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0029] In the description of this invention, the use of terms such as "upper," "lower," "inner," and "outer" to indicate orientation or positional relationship is based on the orientation or positional relationship shown in the accompanying drawings and is only for the convenience of describing the invention and simplifying the description. It does not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion, but rather may include other units not explicitly listed or inherent to these products or devices.

[0030] Combination Figures 1 to 5 As shown in the embodiment, an electrophysiological information acquisition interface device based on transwell (hereinafter referred to as "the acquisition interface device") is disclosed. The acquisition interface device mainly consists of four parts: a cell porous culture medium seat 1, a PCB signal transmission module 2, a transwell signal acquisition unit 3, and a top cover 4.

[0031] like Figures 3a to 3b As shown, the porous cell culture medium holder 1 mainly includes transwell insertion holes 11, cell culture medium perfusion channels 12, and a base body 13. Multiple transwell insertion holes 11 are present, preferably with shapes and sizes adapted to the outer contour and dimensions of the selected transwell design. In this embodiment, the acquisition interface device has nine circular transwell insertion holes 11, i.e., a nine-hole design. However, in actual use, the number and arrangement of the transwell insertion holes 11 can be set according to application requirements and are not limited to this. The cell culture medium perfusion channel 12 is located at the bottom of the transwell insertion holes 11 and communicates with at least one transwell insertion hole 11. Specifically, it can be configured as one, two, or more cell culture medium perfusion channels as needed. Multiple transwell insertion holes 11 connected to the same cell culture medium perfusion channel 12 can be used to construct organoid culture, multi-organ interaction, and other systems. Different cell culture medium perfusion channels 12 can be used for control experiments. In this embodiment, each transwell can form an organoid model. Three transwell insertion wells 11 in each row are connected by a cell culture medium perfusion channel 12, thus forming a multi-organ interaction model. Different cells can be cultured in the interconnected transwell insertion wells 11, and they can exchange information and substances through the shared cell culture medium perfusion channel 12. Alternatively, each column of transwell insertion wells 11 can be designed to be connected by a single cell culture medium perfusion channel 12. In this embodiment, the cell porous culture plate 1 has a rectangular structure, and the transwell insertion wells 11 are arranged in a rectangular array. In other embodiments, the transwell insertion wells 11 can also be arranged in a circular array or other ways. The cell culture medium perfusion channel 12 can also be flexibly designed according to the arrangement of the transwell insertion wells 11 and the transwell insertion wells 11 that need to be connected. Correspondingly, the cell porous culture plate 1 can also be designed as a circle or other structures. In this embodiment, the porous cell culture medium 1 can be customized using 3D printing of transparent resin material. In other embodiments, the porous cell culture medium 1 can also be made of biocompatible transparent materials such as PS (polystyrene) and PMMA (polymethyl methacrylate).

[0032] like Figure 4 As shown, the PCB signal transmission module 2 specifically includes components such as a PCB board 21, dual-row through-hole females 22, dual-row through-hole pins 23, and gold finger electrodes 24. The PCB board 21 can be customized according to the cell porous culture medium holder 1. Multiple mounting holes 25 on the PCB board 1 have shapes, sizes, numbers, and arrangements that correspond to the transwell insertion holes 31 in the cell porous culture medium holder 1. Each mounting hole 25 on the PCB board 1 has a pre-drilled pad insertion hole 26 next to it, and the dual-row through-hole females 22 are inserted into the pad insertion holes 26. The dual-row through-hole females 22 and dual-row through-hole pins 23 are compatible components and can use directly purchased standard parts. In this embodiment, the mounting holes 25 are circular through holes. Considering the PCB board area limitation, 12-pin dual-row through-hole females 22a are used next to the three mounting holes 25 in the middle row, and 20-pin dual-row through-hole females 22b are used next to the six mounting holes 25 on both sides. Gold finger electrodes 24 are disposed on both sides of the PCB board 21 and have pins that correspond to the corresponding dual-row through-hole females 22. Therefore, any pin of the dual-row through-hole female 22 can be connected to the corresponding pin of the gold finger electrode 24 through the pad insertion hole 26. The gold finger electrodes 24 can be used to connect external electrophysiological instruments. A signal transmission channel is formed by the sequentially connected dual-row through-hole pins 23, dual-row through-hole females 22 and gold finger electrodes 24. Therefore, this embodiment has two signal transmission channels, one with 12 channels and the other with 20 channels. After the dual-row through-hole females 22 are inserted into the pad insertion hole 26, they can be soldered to ensure that the gold finger electrodes 24 corresponding to both sides of the PCB board 21 in each dual-row through-hole female 22 are conductive and do not cause crosstalk. The PCB signal transmission module 2 can be fixed to the base body 13 of the cell porous culture medium pedestal 1 by nano double-sided adhesive, or it can be fixed to the cell porous culture medium pedestal 1 by other biocompatible methods.

[0033] like Figure 5As shown, each transwell signal acquisition unit 3 mainly consists of three components: a transwell 31, a clip 32, and a flexible microelectrode array 33. The transwell 31 can be purchased directly as a standard transwell component or customized as needed. The transwell 31 typically has a cup body 311, a neck 312, and a retaining ring 313 connected sequentially from bottom to top. The cup body 311 is primarily used for cell culture; it is usually a cylindrical structure with polypropylene sidewalls and a polycarbonate semi-permeable membrane at the bottom. The pore size of the semi-permeable membrane is approximately 0.4 μm, allowing for material exchange between the cup body and the cell culture medium perfusion channel. The retaining ring 313 has an outer contour larger than the cup body 311 and is mainly used to cooperate with the mounting hole 25 of the PCB signal transmission module 2 to fix the transwell 31 within the transwell insertion hole 11 in the porous cell culture medium holder 1. The snap-fit ​​32 can be customized using 3D printing with transparent resin or other photosensitive materials. It has a rectangular socket 321 and a slot 322. The socket 321 is sized to match the dimensions of the corresponding dual-row straight-through pin header 23, allowing the lower pins of the dual-row straight-through pin header 23 to pass through smoothly. The slot 322 is sized to match the retaining ring 313 on the upper part of the transwell 31, allowing the retaining ring 313 on the upper part of the transwell 31 to fit snugly into the slot 322, forming a snap-fit ​​and thus fixing the snap-fit ​​32 to the transwell 31. After the snap-fit ​​32 is installed, the lower pins of the dual-row straight-through pin header 23 pass through the socket 321 of the snap-fit ​​32 and are inserted into the corresponding dual-row straight-through pin header 22 to achieve electrical connection. After connecting the dual-row straight-through pin header 23 and the transwell 31 together using the snap-fit ​​32, glue can be further applied at the connection between the slot 322 and the retaining ring 313 of the transwell 31 for further fixation. The flexible microelectrode array 33 has a connection hole 331 at one end and an electrode array point 332 at the other end, where each electrode array point 332 represents a signal acquisition channel. The number of signal acquisition channels in the flexible microelectrode array 33 can be designed according to actual needs, as long as it does not exceed the number of signal transmission channels formed by the double-row through-hole pin 22, the double-row through-hole pin 22 and the gold finger electrode 24. In this embodiment, the flexible microelectrode array 33 can be uniformly designed with 10 signal acquisition channels, which is obviously less than the number of the above two types of signal transmission channels. One end of the flexible microelectrode array 33 is inserted into the upper row of pins of the double-row through-hole pin 32 through the connection hole 331 and connected to the signal transmission channel; the other end containing the electrode array point 332 is placed in the cup body 311 of the transwell 31 for collecting electrophysiological information.

[0034] The top cover 4 is mainly used to protect cells from external environmental contamination. It can also be custom-made using 3D printing of transparent resin material and used in conjunction with the porous cell culture medium 1. In other embodiments, the top cover 4 may be omitted depending on the application scenario. In embodiments with the top cover 4, the top cover 4 and the porous cell culture medium 1 are usually made of the same material, such as transparent resin material, PS (polystyrene), PMMA (polymethyl methacrylate), or other biocompatible transparent materials.

[0035] It should be noted that, in this embodiment, the snap-fit ​​32 in the transwell signal acquisition unit 3 serves as a connector. Its main function is, on the one hand, to physically connect and fix the transwell 31 and the double-row through-hole pin 23, so that the double-row through-hole pin 23, the transwell 31, and the flexible microelectrode array 33 in the transwell signal acquisition unit 3 are interconnected to form an integral structure; on the other hand, it can also realize the positioning of the double-row through-hole pin 23, ensuring that the double-row through-hole pin 23 can form a reliable electrical connection with the double-row through-hole pin 22. Therefore, in other embodiments, other forms or structures of connectors can also be used, as long as the above functions can be achieved. Of course, since it does not affect the electrophysiological information acquisition and transmission function, this connector can also be omitted.

[0036] It should also be noted that in this embodiment, the dual-row through-hole female connector 22 and dual-row through-hole male connector 23 are used together, forming an adapter assembly with the pad insertion holes. This assembly electrically connects the signal acquisition channel and the signal transmission channel, allowing the electrophysiological information acquired by the flexible microelectrode array 33 in the transwell signal acquisition unit 3 to be transmitted to an external electrophysiological instrument via the PCB signal transmission module 2. Because of the pluggable nature of the dual-row through-hole female connector 22 and dual-row through-hole male connector 23, the transwell signal acquisition unit 3 and the PCB signal transmission module 2 can be plugged and unplugged, thus allowing for flexible replacement of the transwell signal acquisition unit 3 as needed. In other embodiments, other types of adapter assemblies can also be used, as long as they can achieve the basic function of electrically connecting the signal acquisition channel and the signal transmission channel.

[0037] Based on the above description, it can be seen that the acquisition interface device is based on transwell and chip microchannel technology to perform dynamic tissue culture and organ model construction. It also integrates a PCB board and a flexible microelectrode array, which can not only perform common cell detection, such as cell liveness and death labeling and staining, protein detection, etc., but also use the electrophysiological information collected during cell or tissue culture for analysis and detection.

[0038] The main steps for using the transwell-based pluggable electrophysiological information acquisition interface device disclosed in the embodiments are described below:

[0039] Step 1, device assembly: Assemble individual transwells 31 with snap-fit ​​32, flexible microelectrode array 33 and double-row straight-insertion pins 23 to form a whole; then insert transwells 31 into the transwell insertion holes 11 of the porous culture medium seat 1; then insert the double-row straight-insertion pins 23 into the corresponding double-row straight-insertion pins 22 on the outer periphery of the transwell insertion holes 11.

[0040] Step 2, chip preprocessing: The entire acquisition interface device can be sterilized by ultraviolet light irradiation or by ethylene oxide sterilization.

[0041] Step 3, cell seeding and culture: Seed the cells to be tested in transwell 31, such as cardiomyocytes, nerve cells, etc.; then place the entire collection interface device in a 37℃ CO2 incubator and let it stand for 12-24 hours to wait for cell growth; then add culture medium to the cell porous culture medium seat 1, which can be pumped into the cell culture medium perfusion channel 12 through an external device. After the injection is completed, cover the top cover 4 for dynamic culture.

[0042] Step 4, Signal Acquisition: Connect the gold finger electrode 24 to the electrophysiological instrument, acquire the electrophysiological information generated by the cells in each transwell 31 through the signal acquisition channel corresponding to each transwell 31, and transmit the electrophysiological information to the electrophysiological instrument through the corresponding signal transmission channel. In this way, the electrophysiological information such as the action potential or local field potential of the cultured cells in the transwell 31 can be observed in real time.

[0043] Step 5, Removable Transwell 31 Replacement: After electrophysiological information analysis is completed, if cells in transwell 31 have died or matured, a robotic arm can be used to clamp both ends of the latch 32, lift it vertically, remove the corresponding transwell signal acquisition unit 3, and then insert a new set of signal acquisition units 31 for automated replacement. Alternatively, the latch 32 and the double-row straight-insertion pin header 23 can be manually removed for manual replacement.

[0044] In summary, when cells are cultured in a transwell, this acquisition interface device can acquire signals through the flexible microelectrode array 33 of the transwell signal acquisition unit 3, and then transmit the electrophysiological signals to the electrophysiological instrument for analysis via the PCB signal transmission module 2. The transwell signal acquisition unit 3 can also be designed as a pluggable type via adapter components (dual-row straight-through pin header 23 and dual-row straight-through pin header 22), allowing for flexible replacement as needed.

[0045] Finally, it should be noted that although the embodiments of the present invention have been described above in conjunction with the accompanying drawings, the present invention is not limited to the specific embodiments and application fields described above. The specific embodiments described above are merely illustrative and instructive, and not restrictive. Those skilled in the art, guided by this specification, can make many other forms without departing from the scope of protection of the claims of the present invention, and all of these are within the scope of protection of the present invention.

Claims

1. A transwell-based electrophysiological information acquisition interface device, characterized in that, It includes a cell porous culture medium holder, a PCB signal transmission module, and several signal acquisition units; The cell porous culture medium holder includes a base body and a plurality of transwell insert holes and at least one perfusion channel arranged within the base body; the perfusion channel is connected to at least one transwell insert hole; The PCB signal transmission module includes a PCB board, several adapter components, and several gold finger electrodes arranged on the PCB board; the PCB board has mounting holes that cooperate with the transwell plug-in holes, and the adapter components are respectively arranged on the outer periphery of each mounting hole and electrically connected to the corresponding gold finger electrodes. The signal acquisition unit is equipped with a transwell and a flexible microelectrode array; the transwell can be inserted into the mounting hole and placed inside the transwell insertion hole; one end of the flexible microelectrode array is electrically connected to the adapter component on the periphery of the mounting hole, and the other end is placed inside the transwell for acquiring electrophysiological information of cells cultured inside the transwell; The adapter assembly includes pad insertion holes disposed around the mounting hole, and matching dual-row through-hole female and dual-row through-hole pins; the dual-row through-hole female is inserted into the pad insertion hole and electrically connected to the corresponding gold finger electrode through the pad insertion hole; the lower pin of the dual-row through-hole pin is inserted into the dual-row through-hole female, and the upper pin is electrically connected to one end of the flexible microelectrode array.

2. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The infusion channel has at least two channels.

3. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The transwell plug-in holes are arranged in a rectangular or circular array; the shape and size of the transwell plug-in holes and mounting holes are adapted to the transwell.

4. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The cell porous culture medium seat is made of a biocompatible transparent material.

5. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The PCB signal transmission module is fixed to the base body of the cell porous culture medium using nano double-sided adhesive.

6. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The transwell in the signal acquisition unit has a cup body, a neck, and a retaining ring connected sequentially from bottom to top; the bottom of the cup body is a semi-permeable membrane, through which the exchange of substances between the cup body and the infusion channel is realized; the retaining ring is used in conjunction with the mounting hole to fix the transwell in the corresponding transwell insertion hole.

7. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, It also includes a cover for use with a cell porous culture medium stand.

8. The electrophysiological information acquisition interface device as described in claim 1, characterized in that, The signal acquisition unit is also equipped with a connector; the transwell, flexible microelectrode array, and dual-row through-hole pin header are fixedly connected through the connector to form a whole.

9. The electrophysiological information acquisition interface device as described in claim 8, characterized in that, The connector has a socket and a slot; the lower pins of the double-row straight pin header pass through the socket and are inserted into the double-row straight pin header nut; the connector is engaged with the transwell through the slot.

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