Electrochemical stimulation device for cell culture and cell scratch

Abstract

The utility model provides an electrochemical stimulation device for cell culture and cell scratches, and belongs to the field of cell activity performance testing. Comprising a culture box with a culture cavity, a cover plate for sealing the culture cavity, a working electrode, an auxiliary electrode, a reference electrode, a bracket and a scratching device, through holes for the working electrode, the auxiliary electrode and the reference electrode to penetrate through the cover plate to enter the culture cavity are formed in the cover plate; the reference electrode is detachably connected with the bracket; the scratching device is located in the culture cavity and detachably connected with one side of the cover plate, and the end, away from the cover plate, of the scratching device can abut against the working electrode. The cell culture device can be used for performing electrochemical potential stimulation while meeting the requirements of cell culture and cell scratching work; the cell culture device is simple in test process and simple and convenient to operate. In addition, the device can be repeatedly used, and experiment loss is reduced.

Description

Technical Field

[0001] The utility model relates to the field of cell viability performance testing, and in particular to an electrochemical stimulation device for cell culture and cell scratching. Background Art

[0002] Cell adhesion and migration play a vital role in tissue repair and wound healing. The interaction between biomaterials and cells, the direction and speed of cell adhesion and migration are of great significance in the study of tissue regeneration. Self-assembled membranes have been used to construct bionic interfaces for culturing cells. When external stimuli change, the surface properties of self-assembled membranes change dynamically, and they have the function of transformation and regulation. This "smart surface" with dynamic functional characteristics is of great significance in the modification of biomaterial surfaces. For example, when a self-assembled membrane with a metal substrate is stimulated by an electrochemical potential, the charged functional groups of the self-assembled membrane can undergo conformational changes, thereby affecting the adhesion and migration of cells cultured on the surface of the self-assembled membrane. Therefore, it is necessary to design an experimental device that can not only culture cells on the self-assembled membrane, observe the adhesion ability of cells, and perform simple tests on cell migration, but also connect an electrochemical instrument to directly electrochemically stimulate the self-assembled membrane. Utility Model Content

[0003] The purpose of the utility model is to provide an electrochemical stimulation device for cell culture and cell scratching, which can solve the technical problems in the above-mentioned background technology.

[0004] In order to solve the above technical problems, the technical solution adopted by the utility model is:

[0005] An electrochemical stimulation device for cell culture and cell scratching, comprising a culture box with a culture cavity, a cover plate for sealing the culture cavity, a working electrode, an auxiliary electrode, a reference electrode, a bracket and a scratching device;

[0006] The cover plate is provided with through holes for the working electrode, the auxiliary electrode and the reference electrode to pass through the cover plate and enter the culture chamber;

[0007] The reference electrode is detachably connected to the bracket;

[0008] The scratching device is located in the culture chamber and is detachably connected to one side of the cover plate, and one end of the scratching device away from the cover plate can abut against the working electrode.

[0009] Furthermore, the working electrode includes a first metal wire and a conductive sheet. The conductive sheet is installed at the bottom of the culture chamber. One end of the first metal wire passes through the through hole and abuts against the conductive sheet.

[0010] Furthermore, the auxiliary electrode is a second metal wire, and the length of the second metal wire in the culture chamber is smaller than the height of the culture chamber.

[0011] Furthermore, the bracket includes a base, a fixing rod and a fixing piece for fixing the reference electrode, one end of the fixing rod is connected to the base, and the fixing piece is detachably connected to the fixing rod.

[0012] Furthermore, the scratching device includes a fixing plate, a connecting rod and a dividing piece. The fixing plate and the dividing piece are respectively installed at two ends of the connecting rod. The fixing plate is detachably connected to the cover plate.

[0013] Preferably, a first groove is formed at the bottom of the dividing piece, and a second groove is formed along the height direction of the dividing piece, and the first groove and the second groove are connected.

[0014] Preferably, an electrochemical stimulation device for cell culture and cell scratching further includes a fixing seat, the fixing seat is provided with a third groove for fixing the culture box, a bracket is mounted on the fixing seat, and the bracket is a four-degree-of-freedom robotic arm.

[0015] Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects:

[0016] The utility model provides an electrochemical stimulation device for cell culture and cell scratching, comprising a culture box with a culture cavity, a cover plate for sealing the culture cavity, a working electrode, an auxiliary electrode, a reference electrode, a bracket and a scratching device; the cover plate is provided with through holes for the working electrode, the auxiliary electrode and the reference electrode to pass through the cover plate and enter the culture cavity; the reference electrode is detachably connected to the bracket; the scratching device is located in the culture cavity and is detachably connected to one side of the cover plate, and the end of the scratching device away from the cover plate can abut against the working electrode.

[0017] The cell culture device can not only meet the needs of cell culture and cell scratching, but also can perform electrochemical potential stimulation; the cell culture device has a simple test process and is easy to operate. In addition, the device can be reused, reducing experimental losses. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the utility model, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the utility model and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying creative work.

[0019] Figure 1 This is a schematic diagram of the structure of an electrochemical stimulation device for cell culture and cell scratching provided by the utility model;

[0020] Figure 2 A schematic diagram of the internal structure of the scratching device provided by the utility model;

[0021] Figure 3 for Figure 1 A magnified view of the structure at center A;

[0022] Figure 4 This is a schematic diagram of the structure in which the working electrode, auxiliary electrode, reference electrode and scratching device provided by the utility model are located inside the culture chamber.

[0023] Icons: 100-culture box; 101-culture chamber; 110-cover plate; 111-through hole; 120-fixing seat; 121-third groove; 130-working electrode; 131-first metal wire; 133-conductive sheet; 150-auxiliary electrode; 170-reference electrode; 180-bracket; 181-base; 183-fixing rod; 185-fixing piece; 187-first fixing part; 189-second fixing part; 190-scratching device; 191-fixing plate; 193-connecting rod; 195-dividing piece; 196-first groove; 197-second groove. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical scheme and advantages of the embodiments of the utility model clearer, the technical scheme in the embodiments of the utility model will be clearly and completely described below in conjunction with the drawings in the embodiments of the utility model. Obviously, the described embodiments are part of the embodiments of the utility model, not all of the embodiments. Generally, the components of the embodiments of the utility model described and shown in the drawings here can be arranged and designed in various different configurations.

[0025] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the present invention to be protected, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1

[0027] Please refer to Figures 1 to 4 As shown, an electrochemical stimulation device for cell culture and cell scratching includes a culture box 100 provided with a culture chamber 101, a cover plate 110 for sealing the culture chamber 101, a working electrode 130, an auxiliary electrode 150, a reference electrode 170, a bracket 180 and a scratching device 190. The cover plate 110 is provided with a through hole 111 for the working electrode 130, the auxiliary electrode 150 and the reference electrode 170 to pass through the cover plate 110 and enter the culture chamber 101. The reference electrode 170 is detachably connected to the bracket 180. The scratching device 190 is located in the culture chamber 101 and is detachably connected to one side of the cover plate 110, and the end of the scratching device 190 away from the cover plate 110 can abut against the working electrode 130.

[0028] When the device is in use, first, the staff needs to stick one end of the working electrode 130 to the bottom of the culture chamber 101. Secondly, place the complete culture medium and cells in the culture chamber 101, and then cover the cover plate 110 on the culture chamber 101. Thirdly, pass the other end of the working electrode 130 through one of the through holes 111 so that it is located outside the culture chamber 101. Next, the ends of the auxiliary electrode 150 and the reference electrode 170 are placed in the culture chamber 101 through another through hole 111, and are located below the page of the complete culture medium, ensuring that the three electrodes do not contact each other. Subsequently, stick the bottom of the bracket 180 to the top surface of the cover plate 110, and fix the end of the reference electrode 170 away from the cover plate 110 on the bracket 180. Then, use hot melt adhesive to seal the multiple through holes 111, so that the working electrode 130, the auxiliary electrode 150 and the reference electrode 170 are all sealed and connected to the cover plate 110. At this time, the device can be placed in a carbon dioxide constant temperature incubator for cell culture.

[0029] To perform potential stimulation, the working electrode 130, the auxiliary electrode 150 and the reference electrode 170 are all connected to an external electrochemical workstation, and then the three electrodes are energized to perform potential stimulation. In this embodiment, the working electrode 130 and the auxiliary electrode 150 are connected to the external electrochemical workstation using a U-shaped terminal to an alligator clip wire, and the working electrode 130 or the auxiliary electrode 150 can be quickly connected through the alligator clip.

[0030] To perform cell scratching, complete culture medium is added to the cell and a fixed potential is applied to culture the cell for 24 hours, and then the cell is washed twice with PBS solution. Next, the scratching device 190 is taken out of the culture chamber 101, and the complete culture medium in the culture chamber 101 is replaced with a culture medium containing <2% FBS or a serum-free culture medium, and finally photographed for observation.

[0031] According to the experimental conditions, the staff can select a culture box 100 with one culture cavity 101 or multiple culture cavities 101, and a cover plate 110 covering one culture cavity 101 or multiple culture cavities 101 at the same time. Both the culture box 100 and the cover plate 110 can be reused.

[0032] Specifically, the working electrode 130 includes a first metal wire 131 and a conductive sheet 133. The conductive sheet 133 can be installed at the bottom of the culture chamber 101 through a patch, and the conductive sheet 133 is a gold-plated silicon sheet. The first metal wire 131 is made of Au material, and its diameter is 0.3 mm. The diameter of the through hole 111 through which the first metal wire 131 passes is slightly larger than the diameter of the first metal wire 131, so as to facilitate the sealing connection between the first metal wire 131 and the cover plate 110. One end of the first metal wire 131 abuts against the surface of the conductive sheet 133. The staff can decide whether to decorate the self-assembly mold on the surface of the conductive sheet 133 according to the experimental needs.

[0033] Furthermore, the auxiliary electrode 150 is a second metal wire made of platinum, and the diameter of the second metal wire is also 0.3 mm. The diameter of the through hole 111 through which the second metal wire passes is also slightly larger than the diameter of the second metal wire, so as to facilitate the sealing connection between the second metal wire and the cover plate 110 .

[0034] It should be noted that since the conductive sheet 133 is located at the bottom of the culture chamber 101 and the three electrodes cannot contact each other, the length of the second metal wire in the culture chamber 101 is less than the height of the culture chamber 101 to avoid contact between the second metal wire and the conductive sheet 133.

[0035] Please refer again Figure 1 and Figure 3 As shown, the bracket 180 includes a base 181, a fixing rod 183 and a fixing member 185 for fixing the reference electrode 170. One end of the fixing rod 183 is connected to the base 181, and the base 181 can enhance the stability of the bracket 180. The fixing member 185 is detachably connected to the fixing rod 183, and the fixing member 185 includes a first fixing portion 187 and a second fixing portion 189, the first fixing portion 187 and the second fixing portion 189 are hinged, and the first fixing portion 187 and the second fixing portion 189 are both composed of two Ω-shaped clips, and the ends of the two adjacent Ω-shaped clips away from the hinge bolt are connected by bolts. The first fixing portion 187 is sleeved on the fixing rod 183, and by changing the distance between the two adjacent Ω-shaped clips, the first fixing portion 187 can be clamped on the fixing rod 183, or can be moved along the height direction of the fixing rod 183, thereby changing the position of the fixing member 185. The second fixing portion 189 is sleeved on the reference electrode 170, and the reference electrode 170 can be fixed or separated from the reference electrode 170 by changing the distance between two adjacent Ω-shaped clips. In addition, by changing the angle between the first fixing portion 187 and the second fixing portion 189, the angle of the reference electrode 170 can also be changed, so that the reference electrode 170 can pass through the through hole 111.

[0036] It should be noted that the reference electrode 170 is an existing product, and its structure will not be described in detail here. The reference electrode 170 is an Ag / AgCl electrode, and the internal reference solution is a KCL solution.

[0037] Please refer to Figure 2 and Figure 4 The scratching device 190 includes a fixing plate 191, a connecting rod 193 and a dividing piece 195. The fixing plate 191 and the middle part of the dividing piece 195 are respectively installed at the two ends of the connecting rod 193. The fixing plate 191 is snap-fitted or bonded to the cover plate 110. When the scratching device 190 is placed in the culture chamber 101, the bottom of the dividing piece 195 abuts against the upper surface of the conductive sheet 133, and the cells cannot enter the area where the dividing piece 195 contacts the conductive sheet 133, which provides necessary conditions for the subsequent cell observation work.

[0038] Preferably, a first groove 196 is provided at the bottom of the partition 195, and a second groove 197 is provided along the height direction of the partition 195, and the first groove 196 and the second groove 197 are connected. The lower end of the first metal wire 131 enters the first groove 196 through the second groove 197. By providing the first groove 196 and the second groove 197, the first metal wire 131 can be limited, ensuring that the first metal wire 131 can contact the conductive sheet 133, and the contact area between the first metal wire 131 and the conductive sheet 133 can be increased.

[0039] Example 2

[0040] The difference between the electrochemical stimulation device for cell culture and cell scratching provided in this embodiment and the electrochemical stimulation device for cell culture and cell scratching provided in the above embodiment is that it also includes a fixing seat 120, and a third groove 121 is provided on the fixing seat 120. The groove wall of the third groove 121 is gap-connected with the box wall of the culture box 100, and the third groove 121 can fix the culture box 100. In addition, the bracket 180 is fixedly installed on the fixing seat 120 (not shown in the figure), and the bracket 180 is an existing damped four-degree-of-freedom mechanical arm, which makes the reference electrode 170 more flexible when used.

[0041] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may be subject to various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An electrochemical stimulation device for cell culture and cell scratching, characterized in that: It comprises a culture box provided with a culture cavity, a cover plate for sealing the culture cavity, a working electrode, an auxiliary electrode, a reference electrode, a bracket and a scratching device; The cover plate is provided with through holes for the working electrode, the auxiliary electrode and the reference electrode to pass through the cover plate and enter the culture chamber; The reference electrode is detachably connected to the bracket; The scratching device is located in the culture chamber and is detachably connected to one side of the cover plate. One end of the scratching device away from the cover plate can abut against the working electrode.

2. The electrochemical stimulation device for cell culture and cell scratching according to claim 1, characterized in that: The working electrode comprises a first metal wire and a conductive sheet. The conductive sheet is installed at the bottom of the culture chamber. One end of the first metal wire passes through the through hole and abuts against the conductive sheet.

3. The electrochemical stimulation device for cell culture and cell scratching according to claim 1, characterized in that: The auxiliary electrode is a second metal wire, and the length of the second metal wire in the culture chamber is smaller than the height of the culture chamber.

4. The electrochemical stimulation device for cell culture and cell scratching according to claim 1, characterized in that: The bracket comprises a base, a fixing rod and a fixing piece for fixing the reference electrode, one end of the fixing rod is connected to the base, and the fixing piece is detachably connected to the fixing rod.

5. The electrochemical stimulation device for cell culture and cell scratching according to claim 1, characterized in that: The scratching device comprises a fixing plate, a connecting rod and a dividing piece, wherein the fixing plate and the dividing piece are respectively mounted at two ends of the connecting rod, and the fixing plate is detachably connected to the cover plate.

6. The electrochemical stimulation device for cell culture and cell scratching according to claim 5, characterized in that: A first groove is formed at the bottom of the partition, and a second groove is formed along the height direction of the partition. The first groove is connected to the second groove.

7. The electrochemical stimulation device for cell culture and cell scratching according to claim 1, characterized in that: It also includes a fixing seat, on which a third groove for fixing the culture box is provided, and the bracket is installed on the fixing seat, and the bracket is a four-degree-of-freedom mechanical arm.

Images