Isolated cell culture electrical stimulation and recording device

Abstract

The invention relates to the technical field of culture, and discloses an in-vitro cell culture electrical stimulation and recording device which comprises four quarter frames, the four quarter frames are mutually combined to form an integral frame, and a connection adjusting assembly capable of adjusting the distance between every two quarter frames is arranged between every two quarter frames. The recording electrode array comprises a 1 / 4 frame, the side surfaces of the two sides of the 1 / 4 frame are each provided with a first inlaying groove, a stimulating electrode is detachably inlaid in each first inlaying groove, and a first fixing assembly used for fixing the stimulating electrode is arranged in each first inlaying groove. A standard plug-in type experiment unit is formed, a user only needs to put the whole device into a standard culture dish, complex electrode arrangement can be completed, and the experiment preparation process is greatly simplified.

Description

Technical Field

[0001] This invention belongs to the field of cell culture technology, and more specifically, relates to an in vitro cell culture electrical stimulation and recording device. Background Technology

[0002] In neuroscience, cardiomyocyte research, and other fields, applying controlled electrical stimulation to isolated cells and simultaneously recording their electrophysiological responses is an important method for studying cell function, network connectivity, and pathological mechanisms.

[0003] Traditional experimental setups typically separate the stimulation electrode from the recording electrode, which leads to problems such as uneven electric field distribution, inaccurate spatial positioning, complex setup, and difficulty in achieving stable and reproducible experiments in standard culture dishes. In addition, the electric field generated by the stimulation electrode may cause large-area interference to the signal of the recording electrode, affecting the quality of the recorded data. To address these issues, we propose an in vitro cell culture electrostimulation and recording device. Summary of the Invention

[0004] To solve the above-mentioned technical problems, the basic concept of the technical solution adopted by the present invention is as follows:

[0005] An in vitro cell culture electrical stimulation and recording device includes four quarter-frames, which are combined to form an integral frame. An adjustable connection assembly is provided between two quarter-frames to adjust the distance between them. Each quarter-frame has a mounting groove on one side, in which a stimulation electrode is detachably mounted. A fixing component for fixing the stimulation electrode is also provided in the mounting groove. A second mounting groove is provided at the bottom of each quarter-frame, in which a microelectrode array (MEA) is detachably mounted. The MEA includes a recording electrode and a reference electrode. A fixing component for fixing the MEA is also provided in the mounting groove.

[0006] In a preferred embodiment of the present invention, the fixing component includes a screw, the inner wall of the inlay groove is provided with a mounting groove, the stimulation electrode is inserted into the mounting groove, the screw is threadedly connected to a threaded hole on one side of the quarter frame, and the stimulation electrode is fixed in the mounting groove by the screw. By setting the screw, it is convenient for personnel to fix the stimulation electrode in the mounting groove.

[0007] In a preferred embodiment of the present invention, the fixing component two includes two screws, and the inner wall of the inlay groove two is provided with a mounting groove two. The microelectrode array MEA is inserted into the mounting groove two. The screw two is threadedly connected to a threaded hole at the bottom of the inner wall of the quarter frame. The microelectrode array MEA is fixed inside the mounting groove two by the screw two. By setting the screw two, it is convenient for personnel to fix the microelectrode array MEA in the mounting groove two.

[0008] In a preferred embodiment of the present invention, positioning grooves are provided on one side of the inner wall of the first mounting groove and at the bottom of the inner wall of the second mounting groove. The positions of the two positioning grooves correspond to the first screw and the second screw, respectively. By setting the positioning grooves, the first screw and the second screw can be inserted into the positioning grooves during use, thereby improving the fixing firmness of the first screw and the second screw.

[0009] In a preferred embodiment of the present invention, the reference electrode is disposed around the recording electrode and is placed as close as possible to the edge wall of the culture dish.

[0010] In a preferred embodiment of the present invention, the connecting adjustment assembly includes an adjustment plate, a T-shaped sliding plate is installed at the bottom of the adjustment plate, a T-shaped sliding groove is provided at the top of the quarter frame, the T-shaped sliding plate is slidably connected to the T-shaped sliding groove, a plurality of positioning plates are installed at the bottom of the inner wall of the T-shaped sliding groove, a fixing groove is provided at the bottom of the T-shaped sliding plate, and the positioning plates are engaged in the fixing groove. By setting the positioning plates, the positioning plates can be engaged in the fixing groove, thereby limiting and fixing the quarter frame and the adjustment plate.

[0011] In a preferred embodiment of the present invention, the positioning plate is arched in shape and is made of elastic steel sheet.

[0012] In a preferred embodiment of the present invention, a plurality of magnetic plates I are installed on both sides of the inner wall of the T-shaped slide, and magnetic plates II are installed on both sides of the T-shaped slide. The magnetic plates I and II are magnetically connected. By setting magnetic plates I and II, magnetic plates I and II can attract each other, thereby improving the stability of the connection and fixation between the quarter frame and the adjustment plate.

[0013] In a preferred embodiment of the present invention, the top of the adjustment plate is provided with a scale, which ensures the accuracy of personnel adjusting the frame specifications.

[0014] Compared with the prior art, the present invention has the following advantages:

[0015] This invention integrates the stimulating electrode and recording electrode array onto a single frame, forming a standardized "plug-in" experimental unit. Users only need to place the entire device into a standard culture dish to complete complex electrode arrangements, greatly simplifying the experimental preparation process. Furthermore, by vertically embedding the four stimulating electrodes on the side, a three-dimensional stimulation environment is created. Compared to traditional bottom-planar electrodes, this design guides current more concentratedly through the horizontal plane where the cell layer is located, which is beneficial for forming a dual electric field with a specific phase difference in space for temporal interference stimulation, thereby stimulating cells more precisely and efficiently. The recording electrode (MEA) is located at the bottom and directly contacts the cell, while the reference electrode is deliberately placed in the cell-free area at the edge. This physical spatial isolation, combined with the lateral vertical arrangement of the stimulating electrodes, structurally minimizes the pathway of the stimulation current directly flowing through the recording electrode.

[0016] This invention sets the frame as four separate structures, which makes it easy for personnel to adjust the overall frame specifications for installing electrodes. This allows it to be used with culture dishes of different sizes, ensuring the accuracy of cell research data, reducing the limitations of the device's use, and the entire device, as a robust physical frame, can withstand repeated sterilization treatments, significantly reducing the consumable costs of long-term experiments.

[0017] The specific embodiments of the present invention will now be described in further detail with reference to the accompanying drawings. Attached Figure Description

[0018] In the attached diagram:

[0019] Figure 1 This is a schematic diagram of the overall structure of the present invention;

[0020] Figure 2 This is a front cross-sectional view of the present invention;

[0021] Figure 3 For the present invention Figure 2 Enlarged structural diagram of section A in the middle;

[0022] Figure 4 For the present invention Figure 2 Enlarged structural diagram of section B in the middle;

[0023] Figure 5 This is a schematic cross-sectional view of the T-shaped groove structure of the present invention.

[0024] Figure 6 For the present invention Figure 5 Enlarged structural diagram of section C;

[0025] Figure 7 For the present invention Figure 5 Enlarged structural diagram of section D in the middle;

[0026] Figure 8 This is a schematic diagram of the T-shaped sliding plate structure of the present invention.

[0027] In the diagram: 1. Quarter frame; 2. Mounting groove one; 3. Stimulation electrode; 4. Mounting groove one; 5. Screw one; 6. Positioning groove; 7. Mounting groove two; 8. Reference electrode; 9. Recording electrode; 10. Screw two; 11. Mounting groove two; 12. T-shaped slide; 13. Adjustment plate; 14. T-shaped slide plate; 15. Fixing groove; 16. Positioning plate; 17. Magnetic plate one; 18. Magnetic plate two; 19. Scale. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below with reference to the accompanying drawings. The following embodiments are used to illustrate the present invention.

[0029] like Figures 1 to 8 As shown, the present invention provides a technical solution: an in vitro cell culture electrical stimulation and recording device, comprising four quarter-frames 1, which are combined to form an integral frame. A connecting adjustment component is provided between two quarter-frames 1 to adjust the distance between them. An inlay groove 2 is provided on both sides of the quarter-frame 1, in which a stimulation electrode 3 is detachably inlaid, and a fixing component 1 for fixing the stimulation electrode 3 is provided. An inlay groove 2 11 is provided at the bottom of the quarter-frame 1, in which a microelectrode array MEA is detachably inlaid. The microelectrode array MEA includes a recording electrode 9 and a reference electrode 8. A fixing component 2 for fixing the microelectrode array MEA is provided in the inlay groove 2 11.

[0030] Specifically, the four stimulation electrodes 3 are symmetrically distributed on the side wall of the frame, and their axes are perpendicular to the bottom surface of the culture dish. This vertical arrangement perpendicular to the bottom surface allows a relatively uniform and directional temporal interference electric field to be formed in the cell growth area of ​​the culture dish after energization, which can effectively act on the adherent cells.

[0031] One-quarter frame 1 is made of insulating material.

[0032] Furthermore, the fixing component includes a screw 5, and the inner wall of the inlay groove 2 is provided with a mounting groove 4. The stimulation electrode 3 is inserted into the mounting groove 4. The screw 5 is threadedly connected to a threaded hole on one side of the quarter frame 1. The stimulation electrode 3 is fixed inside the mounting groove 4 by the screw 5.

[0033] The screw 5 facilitates the fixing of the stimulation electrode 3 in the mounting groove 4.

[0034] Furthermore, the fixing component two includes screw two 10, the inner wall of the inlay groove two 11 is provided with mounting groove two 7, the microelectrode array MEA is inserted into mounting groove two 7, screw two 10 is threadedly connected to the threaded hole opened at the bottom of the inner wall of the quarter frame 1, and the microelectrode array MEA is fixed inside the mounting groove two 7 by screw two 10.

[0035] The screw 10 facilitates the fixing of the microelectrode array (MEA) in the mounting groove 7.

[0036] Furthermore, positioning grooves 6 are provided on one side of the inner wall of mounting groove 1 4 and the bottom of the inner wall of mounting groove 2 7, and the positions of the two positioning grooves 6 correspond to screw 1 5 and screw 2 10 respectively.

[0037] By setting the positioning groove 6, screw 5 and screw 10 can be inserted into the positioning groove 6 during use, thereby improving the fixing firmness of screw 5 and screw 10.

[0038] Furthermore, the reference electrode 8 is positioned around the recording electrode 9, and is placed as close as possible to the edge of the culture dish. It is recommended to avoid cell coverage in the area where the reference electrode 8 is located during use to ensure that it can stably provide a clean reference potential and effectively reduce the impact of stimulation artifacts and common-mode noise on the recording signal. The recording electrode 9 is arranged in a high-density matrix, which can simultaneously monitor the electrophysiological activity of multiple sites within the culture area.

[0039] Furthermore, the connecting adjustment assembly includes an adjustment plate 13, a T-shaped sliding plate 14 is installed at the bottom of the adjustment plate 13, a T-shaped sliding groove 12 is opened at the top of the quarter frame 1, the T-shaped sliding plate 14 is slidably connected to the T-shaped sliding groove 12, a plurality of positioning plates 16 are installed at the bottom of the inner wall of the T-shaped sliding groove 12, a fixing groove 15 is opened at the bottom of the T-shaped sliding plate 14, and the positioning plates 16 are snapped into the fixing groove 15;

[0040] The positioning plate 16 can be inserted into the fixing groove 15 by setting the positioning plate 16, thereby limiting and fixing the quarter frame 1 and the adjustment plate 13.

[0041] Furthermore, the positioning plate 16 is arched in shape and is made of elastic steel sheet.

[0042] Furthermore, multiple magnetic plates 17 are installed on both sides of the inner wall of the T-shaped slide 12, and magnetic plates 18 are installed on both sides of the T-shaped slide plate 14. The magnetic plates 17 and 18 are magnetically connected.

[0043] By setting up magnetic plate 17 and magnetic plate 18, magnetic plate 17 and magnetic plate 18 can be attracted to each other, thereby improving the stability of the connection and fixation between the quarter frame 1 and the adjustment plate 13.

[0044] Furthermore, the top of the adjustment plate 13 is provided with a scale 19;

[0045] Among them, setting a scale of 19 ensures the accuracy of personnel adjusting the frame specifications.

[0046] The operating principle of an in vitro cell culture electrical stimulation and recording device is as follows: In use, cells are first seeded in a culture dish containing the device of this invention. The cells mainly adhere to the wall in the microelectrode array region at the bottom. The stimulation electrodes 3 on the sides are connected to two or more independent external electrical stimulation signal generators via wires. The recording electrodes 9 and reference electrodes 8 at the bottom are connected to an external high-impedance amplifier and data acquisition system via a flexible circuit board or probe card. External devices apply alternating current signals with different phases to the side electrodes to generate a time-domain interference electric field. Simultaneously, the local field potential signals generated by the cells are recorded through the microelectrode array at the bottom.

[0047] After the experiment, the entire apparatus can be removed from the petri dish for thorough cleaning and sterilization in preparation for the next use.

[0048] When the frame is used in culture dishes of different sizes, the four individual quarter-frames 1 can be directly adjusted. Pulling a quarter-frame 1 causes multiple positioning plates 16 to move, disengaging the positioning plate 16 at the current position of the T-shaped slide plate 14 from the fixing groove 15, and causing the positioning plate 16 at the designated position to re-enter the fixing groove 15. At this time, the four individual quarter-frames 1 separate from each other, creating a certain distance, thereby adjusting the size of the overall frame. This allows it to be used with culture dishes of different sizes. Through the above structure, the frame is set as a four-part structure, which facilitates the adjustment of the size of the overall frame for installing electrodes. This makes it suitable for culture dishes of different sizes, ensuring the accuracy of research cell data and reducing the limitations of the device's use.

Claims

1. An ex vivo cell culture electrical stimulation and recording device comprising four quarter frames (1), characterized in that, Four quarter frames (1) are combined to form a whole frame, and a connection adjusting assembly is arranged between two of the quarter frames (1) to adjust the distance between the two quarter frames (1). A side surface of each of the quarter frames (1) is provided with an inlaid groove (2), and a stimulating electrode (3) is detachably inlaid in the inlaid groove (2). A fixing assembly (1) is arranged in the inlaid groove (2) to fix the stimulating electrode (3). An inlaid groove (11) is arranged at the bottom end of the quarter frame (1), and a micro electrode array (MEA) is detachably inlaid in the inlaid groove (11). The micro electrode array (MEA) comprises a recording electrode (9) and a reference electrode (8). A fixing assembly (2) is arranged in the inlaid groove (11) to fix the micro electrode array (MEA).

2. The device for extracorporeal cell culture, electric stimulation and recording according to claim 1, characterized in that, The fixing assembly (1) comprises a screw (5), and an installation groove (4) is arranged in the inner wall of the inlaid groove (2). The stimulating electrode (3) is clamped into the installation groove (4), and the screw (5) is threadedly connected with a threaded hole arranged on one side of the quarter frame (1). The stimulating electrode (3) is fixed in the installation groove (4) by the screw (5).

3. The device for extracorporeal cell culture, electric stimulation and recording according to claim 1, characterized in that, The fixing assembly (2) comprises a screw (10), and an installation groove (7) is arranged in the inner wall of the inlaid groove (11). The micro electrode array (MEA) is clamped into the installation groove (7), and the screw (10) is threadedly connected with a threaded hole arranged at the bottom end of the inner wall of the quarter frame (1). The micro electrode array (MEA) is fixed in the installation groove (7) by the screw (10).

4. The device for extracorporeal cell culture electric stimulation and recording according to claim 3, characterized in that, A positioning groove (6) is arranged in one side of the inner wall of the installation groove (4) and at the bottom end of the inner wall of the installation groove (7).

5. The device for extracorporeal cell culture, electric stimulation and recording according to claim 1, characterized in that, The reference electrode (8) is arranged at the periphery of the recording electrode (9).

6. The device for extracorporeal cell culture electric stimulation and recording according to claim 1, characterized in that, The connection adjusting assembly comprises an adjusting plate (13), and a T-shaped sliding plate (14) is arranged at the bottom end of the adjusting plate (13). A T-shaped sliding groove (12) is arranged at the top end of the quarter frame (1), and the T-shaped sliding plate (14) is slidably connected with the T-shaped sliding groove (12). A plurality of positioning plates (16) are arranged at the bottom end of the inner wall of the T-shaped sliding groove (12). A fixing groove (15) is arranged at the bottom end of the T-shaped sliding plate (14), and the positioning plates (16) are clamped in the fixing groove (15).

7. The device for extracorporeal cell culture electrical stimulation and recording according to claim 6, characterized in that, The positioning plates (16) are in the shape of an arch, and are made of elastic steel sheets.

8. The device for extracorporeal cell culture electric stimulation and recording according to claim 6, characterized in that, A plurality of magnetic plates (17) are arranged at both sides of the inner wall of the T-shaped sliding groove (12), and a magnetic plate (18) is arranged at both sides of the T-shaped sliding plate (14). The magnetic plates (17) and the magnetic plate (18) are magnetically connected.

9. The device for extracorporeal cell culture electric stimulation and recording according to claim 1, characterized in that, A scale (19) is arranged at the top end of the adjusting plate (13).

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