Electrical stimulator device for primary cultured neurons

Abstract

An electrical stimulator device for primary cultured neurons, the electrical stimulator device for primary cultured neurons comprising a support (100) and platinum wire electrodes (200), wherein a lower end of the support (100) can extend into a container where neurons are planted; the support (100) is provided with two fixing members (110) arranged in parallel; and two platinum wire electrodes (200) are provided, one end of each platinum wire electrode (200) being connected to an external power supply, the other end of each platinum wire electrode (200) being fixed on a respective fixing member (110), and the parts of the two platinum wire electrodes (200) in contact with the neurons being arranged in parallel.

Description

Electrical stimulator device for primary cultured neurons

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on December 11, 2023, with application number 202323367255.5. The entire contents of the above application are incorporated by reference into this application. Technical Field

[0002] The present application relates to the technical field of biological experimental equipment, for example, to an electrical stimulator device for primary cultured neurons. Background Art

[0003] Using controlled, exogenous means to excite neurons is the most common approach in neuroscience. Common methods for stimulating neurons include extracellular potassium stimulation, electrical stimulation, and optogenetic stimulation, each with its own advantages and disadvantages.

[0004] The simplest method for neuronal stimulation is to increase the potassium ion concentration in the extracellular fluid. According to the principle of action potential generation, in the normal resting state, neurons maintain a state of high potassium inside the cells and high sodium outside the cells. The difference in sodium / potassium ion concentrations inside and outside the cells is an important factor in maintaining the resting membrane potential of neurons. During the experiment, increasing the extracellular potassium concentration can reduce the potassium ion concentration gradient inside and outside the neurons, making the neurons more susceptible to depolarization. Increasing the potassium concentration in the extracellular fluid is easy to operate and does not require additional equipment support, so it has been used in many studies. However, its disadvantage is that high potassium stimulation of neurons is often considered to be non-physiological neural excitation, and high potassium stimulation can cause changes in the neuronal transcriptome, which also restricts its widespread use.

[0005] Optogenetic stimulation involves expressing a light-sensitive protein called a channel in vivo or in cultured neurons through viral infection. Laser stimulation then triggers the channel to open, thereby exciting the neurons. Its advantages lie in its ability to operate at the somatic and cellular levels, but its limitations lie in the side effects of overexpressing the channel protein and the limited range of stimulation.

[0006] Electrical stimulation is the method that is closest to physiological stimulation. Its stimulation intensity and frequency can be flexibly adjusted according to the purpose of the experiment. For example, when a weaker 10Hz is used to stimulate neurons, the neurons can produce a more physiological electrical activity response; while at a stronger 40Hz or 80Hz, the neurons produce macroendocytosis. Therefore, the stimulation mode can be changed to study the function of neurons in different scenarios. In vitro electrical stimulation devices usually use concentric electrodes as terminal stimulators. The stimulation range of this type of electrical stimulation device is small, and due to structural limitations, it cannot stimulate neurons planted in the well plate.

[0007] Summary of the Invention

[0008] The present application discloses an electrical stimulator device for primary cultured neurons, comprising: a bracket, the lower end of which can extend into a container in which neurons are implanted, and the bracket has two parallel fixed parts; two platinum wire electrodes, one end of each of the platinum wire electrodes is connected to an external power supply, and the other end of each of the platinum wire electrodes is fixed to the fixed part, and the parts of the two platinum wire electrodes that contact the neurons are arranged in parallel.

[0009] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become obvious from the description below, or will be learned through practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] FIG1 is a schematic structural diagram of an electrical stimulator device for primary cultured neurons according to Example 1 of the present application;

[0011] FIG2 is a schematic structural diagram of an electrical stimulator device for primary cultured neurons according to Example 2 of the present application.

[0012] Reference numerals: 100 , bracket; 110 , fixing member; 111 , first transverse fixing portion; 112 , second transverse fixing portion; 113 , longitudinal fixing portion; 120 , connecting member; 200 , platinum wire electrode. DETAILED DESCRIPTION

[0013] In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. In addition, features defined as "first" or "second" may explicitly or implicitly include one or more of such features, and are used to distinguish and describe features, without distinction of order or importance. In the description of the present application, unless otherwise specified, "multiple" means two or more.

[0014] In the description of this application, it should be noted that, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "connected" should be understood in a broad sense. For example, they can refer to fixed connections, detachable connections, or integral connections; mechanical connections or electrical connections; direct connections or indirect connections through an intermediate medium; and internal connections between two components. Those skilled in the art will understand the specific meanings of the above terms in this application based on the specific circumstances.

[0015] In certain experimental scenarios, such as the preparation of primary neuronal electron microscopy samples, neurons need to be planted directly on the well plate without being removed. In subsequent experiments, neurons need to be stimulated exogenously to excite them based on different experimental objectives. High potassium stimulation (non-physiological), optogenetics (protein overexpression system, too small stimulation range), and concentric electrode stimulators (too small stimulation range and cannot extend into the well plate) are all unsuitable for this scenario due to their respective limitations.

[0016] In summary, there is an urgent need to propose a stimulator device that can be inserted into a container where neurons are implanted and can achieve large-scale stimulation.

[0017] In view of the above situation, an embodiment of the present application provides an electrical stimulator device for primary cultured neurons.

[0018] The structure of the electrical stimulator device for primary cultured neurons according to an embodiment of the present application will be described below with reference to FIG1-FIG2 .

[0019] The present application discloses an electrical stimulator device for primary cultured neurons, as shown in Figures 1 and 2. The electrical stimulator device for primary cultured neurons includes a support 100 and a platinum wire electrode 200. The lower end of the support 100 can be inserted into a container in which neurons are planted. The support 100 has two parallel fixed members 110. There are two platinum wire electrodes 200, one end of each platinum wire electrode 200 is connected to an external power source, and the other end is fixed to the fixed member 110. The portions of the two platinum wire electrodes 200 that contact the neurons are arranged in parallel. It is understood that in actual operation, after the platinum wire electrodes 200 are fixed to the two fixed members 110, the entire support 100 is inserted into the container in which neurons are planted, and then the other ends of the platinum wire electrodes 200 are connected to the external power source. The portions of the two platinum wire electrodes 200 that contact the neurons are arranged in parallel. A rectangular electric field can be generated between the two platinum wire electrodes 200, which can stimulate all neurons in the electric field area, thereby achieving the function of applying electrical stimulation to neurons in a large range.

[0020] In some embodiments, as shown in Figures 1-2, each fixing member 110 includes a first transverse fixing portion 111, a second transverse fixing portion 112, and a longitudinal fixing portion 113. The first transverse fixing portion 111 is used to fix the portion of the platinum wire electrode 200 that contacts neurons, and the second transverse fixing portion 112 is used to fix the portion of the platinum wire electrode 200 that connects to an external power source. The ends of the longitudinal fixing portion 113 are respectively connected to the first transverse fixing portion 111 and the second transverse fixing portion 112. It will be understood that in actual experiments, the end of the platinum wire electrode 200 connected to the external power source is fixed to the second transverse fixing portion 112, and the other end of the platinum wire electrode 200 is then fixed to the first transverse fixing portion 111. The presence of the longitudinal fixing portion 113 ensures that the platinum wire electrode 200 remains straight and connected to the external power source, thereby ensuring that the two platinum wire electrodes 200 can stably generate an electric field and apply electrical stimulation to neurons over a large range.

[0021] In some embodiments, the platinum wire electrode 200 is fastened to the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113 via fixing wires. It will be appreciated that securing the platinum wire electrode 200 to the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113 via fixing wires ensures the stability of the platinum wire electrode 200 on the bracket 100, thereby preventing the platinum wire electrode 200 from falling off the bracket 100 during an experiment.

[0022] In some embodiments, multiple fixing wires are provided, spaced apart along the extending direction of the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113. It will be appreciated that the multiple fixing wires provided on each of the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113 ensure the stability of the platinum wire electrode 200 on the bracket 100, thereby preventing the platinum wire electrode 200 from falling off the bracket 100 during an experiment.

[0023] In some embodiments, the platinum wire electrode 200 is bonded to the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113. It is understood that, compared to using a fixing wire to tie the platinum wire electrode 200 to the first transverse fixing portion 111, the second transverse fixing portion 112, and the longitudinal fixing portion 113, the bonding connection method is more convenient to operate and has a stronger connection reliability.

[0024] In some embodiments, as shown in FIG1 , the first transverse fixing portion 111 and the second transverse fixing portion 112 are located on either side of the longitudinal fixing portion 113 along its thickness direction. It will be appreciated that, in actual operation, since the first transverse fixing portion 111 and the second transverse fixing portion 112 are located on either side of the longitudinal fixing portion 113 along its thickness direction, after the stent 100 is inserted into the container in which the neurons are implanted, the second transverse fixing portion 112 can be located outside the container, making it easier to connect the platinum wire electrode 200 to an external power source.

[0025] In some embodiments, as shown in Figures 1 and 2, the stent 100 further includes a connector 120, the ends of which are respectively connected to the two fixing members 110. It is understood that the stent 100 connects the two fixing members 110 together through the connector 120, making it easier for the user to insert the stent 100 into the container where the neurons are implanted.

[0026] In some embodiments, as shown in Figures 1 and 2, there are multiple connectors 120, and the multiple connectors 120 are arranged at intervals. It is understood that the multiple connectors 120 can improve the connection stability of the entire bracket 100.

[0027] In some embodiments, the multiple connectors 120 and the two fixing members 110 are integrally formed. It will be appreciated that integrally forming the multiple connectors 120 and the two fixing members 110 facilitates the manufacture of the bracket 100 and improves the connection stability between the connectors 120 and the fixing members 110. In the embodiments of the present application, the multiple connectors 120 and the two fixing members 110 can be directly manufactured via 3D (3-dimensional) printing or injection molding.

[0028] In some embodiments, the stent 100 is a plastic part. It is understood that the stent 100 being a plastic part can, on the one hand, avoid contamination of neurons, and on the other hand, has a low manufacturing cost and is lightweight, making it more convenient to use. Of course, it should be noted that in other embodiments of the present application, the stent 100 can also be made of other materials.

[0029] Example 1:

[0030] As shown in Figure 1, the electrical stimulator device for primary cultured neurons includes a bracket 100 and a platinum wire electrode 200. Each fixing member 110 includes a first transverse fixing portion 111, a second transverse fixing portion 112, and a longitudinal fixing portion 113. The first transverse fixing portion 111 is used to fix the portion of the platinum wire electrode 200 that contacts the neurons, and the second transverse fixing portion 112 is used to fix the portion of the platinum wire electrode 200 that connects to the external power supply. The two ends of the longitudinal fixing portion 113 are respectively connected to the first transverse fixing portion 111 and the second transverse fixing portion 112. The first transverse fixing portion 111 and the second transverse fixing portion 112 are located on either side of the longitudinal fixing portion 113 along its thickness direction.

[0031] Example 2:

[0032] As shown in FIG2 , the electrical stimulator device for primary cultured neurons includes a support 100 and a platinum wire electrode 200. Each fixing member 110 includes a first transverse fixing portion 111, a second transverse fixing portion 112, and a longitudinal fixing portion 113. The first transverse fixing portion 111 is used to fix the portion of the platinum wire electrode 200 that contacts the neurons, and the second transverse fixing portion 112 is used to fix the portion of the platinum wire electrode 200 that connects to the external power supply. The ends of the longitudinal fixing portion 113 are respectively connected to the first transverse fixing portion 111 and the second transverse fixing portion 112. The first transverse fixing portion 111 and the second transverse fixing portion 112 are located on the same side of the longitudinal fixing portion 113 along its thickness direction.

[0033] Throughout this specification, references to "some embodiments," "other embodiments," and the like indicate that the features, structures, materials, or characteristics described in connection with that embodiment or example are included in at least one embodiment or example of the present application. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the described features, structures, materials, or characteristics may be combined in any suitable manner in any one or more embodiments or examples.

Claims

1. An electrical stimulator device for primary cultured neurons, comprising: A bracket (100), the lower end of the bracket (100) can extend into a container in which neurons are planted, and the bracket (100) has two fixing members (110) arranged in parallel; Platinum wire electrodes (200), there are two platinum wire electrodes (200), one end of each platinum wire electrode (200) is connected to an external power supply, the other end of each platinum wire electrode (200) is fixed on the fixing member (110), and the parts of the two platinum wire electrodes (200) in contact with the neurons are arranged in parallel.

2. The electrical stimulator device for primary cultured neurons according to claim 1, wherein, Each of the fixing members (110) includes: A first horizontal fixing portion (111) for fixing the part of the platinum wire electrode (200) in contact with the neurons; A second horizontal fixing portion (112) for fixing the part of the platinum wire electrode (200) connected to the external power supply; A longitudinal fixing portion (113), and both ends of the longitudinal fixing portion (113) are respectively connected to the first horizontal fixing portion (111) and the second horizontal fixing portion (112).

3. The electrical stimulator device for primary cultured neurons according to claim 2, wherein, The platinum wire electrode (200) is tied to the first horizontal fixing portion (111), the second horizontal fixing portion (112), and the longitudinal fixing portion (113) by fixing wires.

4. The electrical stimulator device for primary cultured neurons according to claim 3, wherein, There are multiple fixing wires, and the multiple fixing wires are arranged at intervals along the extending directions of the first horizontal fixing portion (111), the second horizontal fixing portion (112), and the longitudinal fixing portion (113).

5. The electrical stimulator device for primary cultured neurons according to claim 2, wherein, The platinum wire electrode (200) is adhered to the first horizontal fixing portion (111), the second horizontal fixing portion (112), and the longitudinal fixing portion (113).

6. The electrical stimulator device for primary cultured neurons according to claim 2, wherein, The first horizontal fixing portion (111) and the second horizontal fixing portion (112) are located on both sides of the longitudinal fixing portion (113) along its thickness direction.

7. The electrical stimulator device for primary cultured neurons according to claim 1, wherein, The bracket (100) further includes a connecting member (120), and both ends of the connecting member (120) are respectively connected to the two fixing members (110).

8. The electrical stimulator device for primary cultured neurons according to claim 7, wherein, There are multiple connecting members (120), and the multiple connecting members (120) are arranged at intervals.

9. The electrical stimulator device for primary cultured neurons according to claim 7, wherein, The multiple connecting members (120) and the two fixing members (110) are integrally formed parts.

10. The electrical stimulator device for primary cultured neurons according to claim 1, wherein, The bracket (100) is a plastic part.

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