Cell stimulation device

Abstract

The invention discloses a cell stimulation device, which comprises a mechanical stimulation unit, a cell clamping unit and a cell placement unit, and is characterized in that the mechanical stimulation unit and the cell clamping unit are mounted on a supporting bottom plate through a fine adjustment connecting plate, a three-way displacement fine adjustment platform and a three-way displacement coarse adjustment platform; and a piezoelectric actuator of the mechanical stimulation unit is rotationally mounted on the fine adjustment connecting plate through a clamping piece. The micro-needle tip can be accurately moved to the surface of a cell through micron-level positioning and movement, micron-level vibration stimulation on the cell is realized based on reciprocating motion of the piezoelectric actuator, and the displacement and frequency of cell pressing can be regulated and controlled by the piezoelectric driving power supply; low-frequency constant-pressure and high-frequency vibration pressing and vibration stimulation with multiple superposed frequencies can be realized; the cells can be clamped and fixed in a negative pressure suction mode of the other glass microneedle, so that the cells can be kept stable when being subjected to mechanical stimulation, and the cells are ensured not to move after being subjected to mechanical stimulation in the culture dish.

Description

Technical Field

[0001] The present invention relates to the intersectional field of medicine and engineering, and in particular to a cell stimulation device. Background Art

[0002] As the basic unit of life activities, cells contain a lot of important information related to life. The study of cells has a significant role in promoting life sciences, physical chemistry and bionics. At present, the research and work on cells are mostly focused on the structure of cells and chemical drug analysis, while there are few studies on the physical properties of cells, especially the study of the related properties of cells under mechanical stimulation. The main reason is that there are great technical difficulties in applying mechanical stimulation at the cellular level, and there are great challenges in the accuracy and stability of related equipment. Summary of the invention

[0003] Purpose of the invention: The present invention aims to provide a cell stimulation device capable of applying mechanical stimulation to the cell membrane surface while clamping and fixing cells.

[0004] Technical solution: The cell stimulation device described in the present invention includes a mechanical stimulation unit, a cell clamping unit and a cell placement unit. The mechanical stimulation unit and the cell clamping unit are rotatably mounted on a three-way displacement fine adjustment platform through a fine adjustment connecting plate. The three-way displacement fine adjustment platform is mounted on a supporting base plate through a three-way displacement coarse adjustment platform. The mechanical stimulation unit includes a piezoelectric actuator, a hexagonal stud, a thread conversion head, a microneedle clamp and a glass microneedle. The piezoelectric actuator is rotatably mounted on the fine adjustment connecting plate through a clamp.

[0005] Furthermore, the piezoelectric actuator includes a housing, a bottom fixing part, a piezoelectric ceramic stack, a preload spring and a rivet. The rivet and the piezoelectric ceramic stack are sequentially sleeved in the housing, the preload spring is sleeved on the rivet, and the bottom fixing part is rotatably fixed to the bottom of the housing. The expansion and contraction of the piezoelectric actuator drives the glass microneedle to achieve reciprocating motion at the micron level and at any frequency. The tip of the glass microneedle applies mechanical stimulation to the cell controlled by the piezoelectric actuator. The displacement of the pressed cell and the pressing frequency can be regulated by the driving power supply of the piezoelectric actuator, thereby applying accurate and stable mechanical stimulation to the cell.

[0006] Furthermore, the movable end of the rivet of the piezoelectric actuator is connected to the hexagonal stud.

[0007] Furthermore, the microneedle clamp includes a microneedle clamping connector, a microneedle fastener and a microneedle conical fixing member.

[0008] Furthermore, the cell clamping unit includes a glass microneedle, a two-way valve and a fixing box. The air hole of the two-way valve is connected to a negative pressure device, so that the tip of the glass microneedle generates negative pressure suction. The fixing box is fixed on the three-way displacement fine adjustment platform through a fine adjustment connecting plate. In order to avoid irregular movement of cells under mechanical stimulation, the cells are clamped under the glass microneedle by negative pressure suction, that is, a stable package is formed between the cell membrane and the glass microneedle through the cell clamping part, thereby effectively fixing the cells and facilitating accurate mechanical stimulation of the cells.

[0009] Furthermore, the fine-tuning connecting plate is provided with a limiting groove.

[0010] Furthermore, the limiting groove is an arc-shaped limiting groove, and the center angle corresponding to the arc is 90°.

[0011] Furthermore, the minimum displacement distance of the three-way coarse adjustment platform in the X / Y / Z axes is 10 microns, and the minimum displacement distance of the three-way fine adjustment platform in the X / Y / Z axes is 2 microns. The microneedle tip can be accurately moved to the cell surface through coarse and fine adjustment of the movement.

[0012] Furthermore, the length of the hexagonal stud is not less than 50 mm.

[0013] Furthermore, the piezoelectric ceramic stack is a rectangular parallelepiped structure formed by stacking multiple layers of piezoelectric ceramics.

[0014] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: 1. The present invention can accurately move the tip of the microneedle to the cell surface through micron-level positioning and movement; 2. The present invention realizes micron-level vibration stimulation of cells based on the reciprocating motion of the piezoelectric actuator, and the displacement and frequency of the cell pressing can be regulated by the piezoelectric driving power supply, which can realize low-frequency constant pressure, high-frequency vibration pressing and vibration stimulation with multiple frequencies superimposed; 3. The present invention can clamp and fix the cells by negative pressure attraction of another glass microneedle, so that the cells can remain stable when mechanically stimulated, ensuring that the cells do not move after being mechanically stimulated in the culture dish. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It is a structural schematic diagram of the present invention;

[0016] Figure 2 Schematic diagram of applying mechanical stimulation to cells and clamping cells with negative pressure;

[0017] Figure 3 is a schematic diagram of the structure of a piezoelectric actuator;

[0018] Figure 4 It is a schematic diagram of the structure of the mechanical stimulation unit;

[0019] Figure 5 The figure is a schematic diagram of the installation of the piezoelectric actuator, the clamping member and the fine-tuning connecting plate;

[0020] Figure 6 It is a schematic diagram of the structure of a piezoelectric ceramic stack;

[0021] Figure 7 Schematic diagram of the structure of the cell clamp unit DETAILED DESCRIPTION

[0022] The present invention will be further described below in conjunction with the accompanying drawings.

[0023] The cell stimulation device of the present invention comprises a mechanical stimulation unit, a cell clamping unit and a cell placement unit. The mechanical stimulation unit is installed on a first supporting base plate 3 through a first X / Y / Z axial three-dimensional displacement coarse adjustment platform 4, a first X / Y / Z axial three-dimensional displacement fine adjustment platform 6 and a fine adjustment connecting plate 5. The tip position of a first glass microneedle 13 of the mechanical stimulation unit is adjusted by adjusting the first X / Y / Z axial three-dimensional displacement coarse adjustment platform 4, the first X / Y / Z axial three-dimensional displacement fine adjustment platform 6 and the fine adjustment connecting plate 5. After the tip of the first glass microneedle 13 is positioned at the cell, the piezoelectric actuator 8 of the mechanical stimulation unit is driven by the principle of the inverse piezoelectric effect to realize the reciprocating motion of the tip of the first glass microneedle 13. The specific displacement and frequency of the reciprocating motion can be regulated by a piezoelectric ceramic driving power supply.

[0024] The mechanical stimulation unit includes a piezoelectric actuator 8, a hexagonal stud 9, a thread conversion head 10, a microneedle clamp 11 and a first glass microneedle 13. The piezoelectric actuator 8 is rotatably mounted on the fine-tuning connecting plate 5 through a clamp 7. The fine-tuning connecting plate 5 is provided with an arc-shaped limit groove, and the arc corresponds to a center angle of 90°. The clamp 7 can rotate around a fixed guide rail along the center of rotation in the fine-tuning connecting plate 5 to adjust the angle of the front end of the piezoelectric actuator 8, thereby adjusting the horizontal deflection angle of the first glass microneedle 13. The designed adjustment angle range is 0 degrees to 90 degrees. The reciprocating displacement generation formula of the first glass microneedle 13 is realized based on the structural design and connection of the piezoelectric actuator. The piezoelectric actuator 8 includes a housing 19, a bottom fixing member 20, a piezoelectric ceramic stack 21, a preload spring 22 and a rivet 23. The rivet 23 and the piezoelectric ceramic stack 21 are sequentially sleeved in the housing 19, the preload spring 22 is sleeved on the rivet 23, and the bottom fixing member 20 is rotatably fixed to the bottom of the housing. The front end of the rivet 23 extends through the housing 19 as a motion transmission part, and the piezoelectric ceramic stack 21 contacts the bottom of the rivet 23 and is preloaded through the bottom fixing member 20. The preload spring 22 is passed through the rivet 23, and the preload spring 22 contacts the housing 19 and the bottom of the rivet 23. There is a threaded connection between the bottom fixing piece 20 and the housing 19. The process of tightening the bottom fixing piece 20 pushes the piezoelectric ceramic stack 21, thereby pushing the rivet 23 so that the preload spring 22 is preloaded. When the piezoelectric ceramic stack generates reciprocating motion, due to the preload of the preload spring 22, the rivet 23 generates the same reciprocating motion, thereby transmitting the reciprocating motion displacement.

[0025] The reciprocating displacement can be transmitted to the first glass microneedle 13 by the rivet 23, and the displacement of the piezoelectric actuator 8 can transmit the reciprocating motion to the first glass microneedle 13 through a series of connections. The housing 19 is clamped and fixed by the clamp 7, the bottom fixing member 20 is fixed to the housing 19 by threaded connection and the piezoelectric actuator 8 is pre-tightened, one end of the rivet 23 is extended by 50 mm through the hexagonal stud 9, and the hexagonal stud 9 is threadedly connected with the microneedle clamping connector 24 through the thread conversion head 10, and the first glass microneedle 13 is passed through the microneedle fastener 25 and the microneedle conical fixing member 26. The microneedle fastener 25 is threadedly connected to the microneedle clamping connector 24. When the two threads are tightened, the microneedle conical fixing member 25 is pushed to clamp the first glass microneedle 13, thereby achieving the fastening of the first glass microneedle 13. After the first glass microneedle 13 is fixed, the reciprocating motion of the rivet 23 can be transmitted to the tip of the first glass microneedle 13 , and the tip can apply a reciprocating mechanical stimulation to the cell 28 .

[0026] Since the displacement generated by a single ceramic piezoelectric body is small, only 1 to 2 microns, it is necessary to use a piezoelectric stacking method to construct a piezoelectric ceramic stack sheet 21 formed by stacking multiple layers of piezoelectric ceramics to generate a larger displacement. Multiple piezoelectric ceramics are stacked to form a piezoelectric stack actuator. The height of the stacked rectangular piezoelectric stack is 25 mm, and the length and width are both 5 mm.

[0027] The cell clamping unit is installed on the second supporting base plate 1 through the second X / Y / Z axial three-dimensional displacement coarse adjustment platform 18, the second X / Y / Z axial three-dimensional displacement fine adjustment platform 17 and the fine adjustment connecting plate 5. The tip position of the second glass microneedle 14 of the cell clamping unit is adjusted by adjusting the second X / Y / Z axial three-dimensional displacement coarse adjustment platform 18, the second X / Y / Z axial three-dimensional displacement fine adjustment platform 17 and the fine adjustment connecting plate 5.

[0028] The cell clamping unit includes a second glass microneedle 14, a second channel 15 and a fixing box 16. The air hole of the second channel 15 is connected to a negative pressure device, so that the tip of the second glass microneedle 14 generates negative pressure attraction. The fixing box 16 is fixed on the three-dimensional displacement fine adjustment platform through the fine adjustment connecting plate 5. The tip position of the second glass microneedle 14 is adjusted by adjusting the first X / Y / Z axial three-dimensional displacement coarse adjustment platform 4, the first X / Y / Z axial three-dimensional displacement fine adjustment platform 5 and the fine adjustment connecting plate 5. After positioning the cell, air is sucked through the through hole of the second channel 15, the air pressure is adjusted, and the cell membrane surface is attracted by negative pressure. The second glass microneedle 14 can be sealed with the cell, and the cell membrane is tightly sealed with the tip of the glass microneedle to achieve a clamping and fixing effect, ensuring that the cell can remain stable under the mechanical stimulation of the first glass microneedle 13.

[0029] Since the cell scale is small, all at the micron level, the tip of the microneedle needs to be positioned and moved to the cell surface, and then mechanical stimulation is applied to the cell through the movement of the piezoelectric actuator. In order to precisely move the microneedle, the coarse adjustment XYZ moving platform and the fine adjustment XYZ moving platform are used to move the microneedle and position it on the cell surface. The minimum displacement distance of the three-way coarse adjustment platform in the X / Y / Z axis is 10 microns, and the minimum displacement distance of the three-way fine adjustment platform in the X / Y / Z axis is 2 microns. The tip of the microneedle can be accurately moved to the cell surface through coarse and fine adjustment of the movement.

[0030] The cell placement unit is composed of a third supporting base plate 2 and a cell culture dish 12 , in which cells 27 with good activity exist.

[0031] The process of using the cell stimulation device of the present invention is as follows:

[0032] Step 1: Use the knobs of the second X / Y / Z axial three-dimensional displacement coarse adjustment platform 18 and the second X / Y / Z axial three-dimensional displacement fine adjustment platform 17 to move the tip of the second glass microneedle 14 to the cell surface.

[0033] Step 2: Suction air from the through hole of the second port 15 to provide negative pressure to slowly absorb, clamp and fix the cells.

[0034] Step 3: Use the knobs of the first X / Y / Z axial three-dimensional displacement coarse adjustment platform 4 and the second X / Y / Z axial three-dimensional displacement fine adjustment platform 6 to move the tip of the first glass microneedle 13 to the cell surface.

[0035] Step 4: Apply an electric field to the piezoelectric ceramic stack 21 through an external piezoelectric ceramic driving power supply, so that the tip of the first glass microneedle 13 produces reciprocating motion, and the displacement and frequency of the reciprocating motion are regulated by the applied voltage amplitude and frequency.

[0036] The present invention can perform micrometer-level mechanical stimulation operations on single cells and can provide a new technical solution for related research on cell response to mechanical stimulation. On the one hand, it can promote the development of related basic fields of life sciences as a new technological invention. On the other hand, it can also promote the intersection of mechanical physics and cell life research and lay the foundation for the research and development of medical and engineering cross-disciplinary equipment.

Claims

1. A cell stimulation device, characterized in that: The invention comprises a mechanical stimulation unit, a cell clamping unit and a cell placement unit. The mechanical stimulation unit and the cell clamping unit are rotatably mounted on a three-dimensional displacement fine adjustment platform via a fine adjustment connecting plate (5). The three-dimensional displacement fine adjustment platform is mounted on a supporting base plate via a three-dimensional displacement coarse adjustment platform. The mechanical stimulation unit comprises a piezoelectric actuator (8), a hexagonal stud (9), a thread conversion head (10), a microneedle clamp (11) and a glass microneedle (13). The piezoelectric actuator (8) is rotatably mounted on the fine adjustment connecting plate (5) via a clamping member (7).

2. The cell stimulation device according to claim 1, characterized in that: The piezoelectric actuator (8) comprises a housing (19), a bottom fixing member (20), a piezoelectric ceramic stack (21), a preload spring (22) and a rivet (23); the rivet (23) and the piezoelectric ceramic stack (21) are sequentially sleeved in the housing (19); the preload spring (22) is sleeved on the rivet (23); and the bottom fixing member (20) is rotatably fixed to the bottom of the housing.

3. The cell stimulation device according to claim 2, characterized in that: The movable end of the rivet of the piezoelectric actuator (8) is connected to the hexagonal stud (9).

4. The cell stimulation device according to claim 1, characterized in that: The microneedle clamp (11) comprises a microneedle clamping connector (24), a microneedle fastener (25) and a microneedle conical fixing member (30).

5. The cell stimulation device according to claim 1, characterized in that: The cell clamping unit comprises a glass microneedle, a two-way port (15) and a fixing box (16); the air hole of the two-way port (15) is connected to a negative pressure device so that the tip of the glass microneedle generates negative pressure suction; the fixing box (16) is fixed on a three-dimensional displacement fine adjustment platform via a fine adjustment connecting plate (5).

6. The cell stimulation device according to claim 1, characterized in that: The fine-tuning connecting plate (5) is provided with a limiting groove.

7. The cell stimulation device according to claim 6, characterized in that: The limiting groove is an arc-shaped limiting groove, and the center angle corresponding to the arc is 90°.

8. The cell stimulation device according to claim 1, characterized in that: The minimum displacement distance of the three-way coarse adjustment platform in the X / Y / Z axis is 10 microns, and the minimum displacement distance of the three-way fine adjustment platform in the X / Y / Z axis is 2 microns.

9. The cell stimulation device according to claim 1, characterized in that: The length of the hexagonal stud (9) is not less than 50 mm.

10. The cell stimulation device according to claim 2, characterized in that: The piezoelectric ceramic stack (21) is a rectangular parallelepiped structure formed by stacking multiple layers of piezoelectric ceramics.

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