CN110872560B - Micro-space electroporation device with adjustable electrode space - Google Patents

Abstract

The invention relates to the technical field of biological experimental equipment, and particularly discloses a micro-spacing electroporation device with adjustable electrode spacing, which comprises a fixed electrode, an adjustable electrode, an electrode seat, a bolt seat and an adjusting bolt; the fixed electrode is arranged on one side of the electrode seat, and the bolt seat is arranged on the other side of the electrode seat; one side part of the adjustable electrode is slidably arranged in the electrode seat, and one side part of the adjustable electrode is in sliding contact with or spaced from one side part of the fixed electrode; the adjusting bolt penetrates through one threaded hole of the bolt seat to be in threaded or fixed connection with the other side portion of the adjustable electrode. The invention has simpler structure and more convenient manufacture by modular design, ensures that the electrode spacing can be adjusted randomly from micron to millimeter, has the adjustment precision exceeding 1 micron, can be integrated into automatic equipment, realizes continuous flow operation of cell electroporation, and improves the experimental efficiency.

Description

A micro-pitch electroporation device with adjustable electrode spacing

technical field

The invention relates to the technical field of biological experiment equipment, in particular to a micro-pitch electroporation device with adjustable electrode spacing.

Background technique

Cell electroporation (Electroporation), also known as electrotransfection technology, is a commonly used method in cell transfection technology. Because the cell membrane is selectively permeable to external substances, the control of cell gene experiments requires the input of specific biological DNA and RNA fragments into the cells. Applying a certain intensity of potential difference on both sides of the cell membrane and continuing for a period of time can generate micropores on the cell membrane and enhance the permeability of the cell membrane. When the cell membrane undergoes electroporation, its permeability will increase instantaneously, so that foreign DNA, protein, drug particles and other substances that cannot pass through the cell membrane under normal circumstances can enter the cell. After the potential difference is removed for a short period of time, the cell membrane can restore itself and become a selective permeability barrier again. Electroporation technology has a wide range of applications in biophysics, molecular biology, clinical medicine and other fields.

The current electroporation devices can be mainly divided into two types: one is a standard electric shock cup with an electrode spacing of 1-4 mm, which is on the millimeter scale, while the cell size is on the micron scale, so the applied voltage is large (generally from several hundred volts to Thousands of volts), and the electric field is not uniform, and the electric field environment of each cell is different, causing the cells near the electrode to die easily, and the cells located at the weaker electric field cannot be transfected by perforation, and the survival rate and transfection efficiency are relatively high. Low, and each operation can only put a small amount of cells and plasmids according to the volume of the standard electric shock cup, and the experimental efficiency is low; the other is a micro-pitch (generally about 1-100um) electroporation device (generally made ), this device requires much lower voltage than the previous device due to the small electrode spacing, so it is safer to use, but because it needs to be made by micro-nano processing technology, the processing technology is relatively complicated, and the existing micro-pitch electroporation The electrode spacing of the device is fixed, which lacks versatility, makes it difficult to adapt to different experimental requirements, and is also difficult to integrate into automated equipment.

Contents of the invention

In view of this, it is necessary to propose a micro-pitch electroporation device with adjustable electrode spacing to solve the above-mentioned shortcomings in the background technology.

To achieve the above object, the present invention takes the following technical solutions:

A micro-pitch electroporation device with adjustable electrode spacing includes a fixed electrode, an adjustable electrode, an electrode seat, a bolt seat, and an adjusting bolt;

The fixed electrode is installed on one side of the electrode seat, and the bolt seat is installed on the other side of the electrode seat; one side of the adjustable electrode is slidably installed in the electrode seat, and one side of the adjustable electrode The part is in sliding contact with or spaced from one side of the fixed electrode; the adjusting bolt passes through a threaded hole of the bolt seat and is threaded or fixedly connected to the other side of the adjustable electrode;

The fixed electrode is used to cooperate with the adjustable electrode so as to apply the voltage required for cell electroporation to the cell fluid in the device;

The adjustable electrode is used to cooperate with the fixed electrode to apply the voltage required for cell electroporation to the cell fluid in the device;

The electrode holder is used to install fixed electrodes and adjustable electrodes, so that the cell fluid forms a cell fluid flow channel in the electrode holder;

The bolt seat is used to screw the adjusting bolt, so that the adjusting bolt moves relative to the electrode seat during rotation;

The adjusting bolt is used to link the adjustable electrode.

Further, one side of the fixed electrode is in the shape of a cylinder, and the other side of the fixed electrode is installed on one side of the electrode holder; the shape of the adjustable electrode is a cylinder; the shape of the fixed electrode is One side portion and one side portion of the adjustable electrode are both embedded in the same through hole of the electrode holder.

Further, the end surface of one side of the fixed electrode is provided with a first inverted tapered concave hole; the end surface of one side of the adjustable electrode is provided with a first tapered frustum; the first inverted tapered The concave hole and the first conical truncated body cooperate with each other or are spaced from each other; the fixed electrode is provided with a first channel, and a channel opening of the first channel is opened on the bottom surface of the first inverted tapered concave hole, so The other channel opening of the first channel is used to input or output cell fluid; the electrode base is provided with a second channel, and one channel opening of the second channel is opened on the inner wall surface of a through hole of the electrode base and is located It is fixedly installed on the side of the end surface of one side of the fixed electrode of the electrode seat, and the other channel opening of the second channel is used for inputting or outputting cell fluid.

Further, a second tapered frustum is provided on the end surface of one side of the fixed electrode; a second inverted tapered concave hole is provided on the end surface of one side of the adjustable electrode; the second inverted tapered The concave hole and the second conical frustum cooperate with each other or are spaced from each other; the fixed electrode is provided with a third channel, and a channel opening of the third channel is opened on the top surface of the second conical frustum, so The other channel port of the third channel is used to input or output cell fluid; the electrode base is provided with a fourth channel, and one channel port of the fourth channel is opened on the inner wall surface of a through hole of the electrode base and is located It is fixedly installed on the side of the end surface of one side of the fixed electrode of the electrode seat, and the other channel opening of the fourth channel is used for inputting or outputting cell fluid.

Further, the taper of the tapered surface of the first inverted tapered concave hole is the same as the taper of the tapered surface of the first tapered frustum.

Further, the taper of the tapered surface of the second inverted tapered concave hole is the same as the taper of the tapered surface of the second tapered frustum.

Further, the adjustment bolt is an M3 bolt; the pitch of the adjustment bolt is 0.5mm; the taper of the conical surface of the first inverted conical concave hole is 1:5; The taper of the conical surface is 1:5.

Further, the adjustment bolt is an M3 bolt; the pitch of the adjustment bolt is 0.5mm; the taper of the conical surface of the second inverted conical concave hole is 1:5; The taper of the conical surface is 1:5.

Further, the fixed electrode is installed on one side of the electrode base through several screws; the bolt base is installed on the other side of the electrode base through several screws.

Further, a first waterproof rubber ring is provided between the cylindrical side wall of the fixed electrode and the through hole wall of the electrode holder; a ring is provided between the cylindrical side wall of the adjustable electrode and the through hole wall of the electrode holder. The second waterproof rubber ring.

The beneficial effects of the present invention are:

In the present invention, the modular design of fixed electrodes, adjustable electrodes, electrode holders, bolt holders and adjustment bolts makes the structure simpler and more convenient to manufacture, so that the electrode spacing can be adjusted arbitrarily between microns and millimeters, and the adjustment accuracy exceeds 1 micron , and can be integrated into automated equipment to realize continuous flow operation of cell electroporation and improve experimental efficiency. The fixed electrode, the adjustable electrode, and the electrode base form a flow channel cavity through which the cell liquid can flow. When the cell liquid mixed with cells and plasmids flows through the flow channel cavity, the electrode is applied to the electrode required for cell electroporation. Voltage, that is, the cell electroporation operation is realized; the present invention is applied in the automation equipment, which can control the continuous flow of the cell fluid through the cavity, and at the same time control the continuous application of voltage to the electrodes, so as to realize the continuous flow operation of the cell electroporation.

Description of drawings

Fig. 1 is a three-dimensional schematic diagram of a micro-pitch electroporation device with adjustable electrode spacing according to the present invention;

2 is a front sectional view of a micro-pitch electroporation device with adjustable electrode spacing according to Embodiment 1 of the present invention;

3 is a partial cross-sectional view of a micro-pitch electroporation device with adjustable electrode spacing according to Embodiment 1 of the present invention;

4 is a front sectional view of a micro-pitch electroporation device with adjustable electrode spacing according to Embodiment 5 of the present invention;

5 is a partial cross-sectional view of a micro-pitch electroporation device with adjustable electrode spacing according to Embodiment 5 of the present invention;

Explanation of reference signs:

Fixed electrode——1; Adjustable electrode——2; Electrode seat——3; Bolt seat——4; Adjusting bolt——5; 21; the first channel - 12; the second channel - 31; the second tapered platform - 13; the second inverted tapered concave hole - 22; the third channel - 14; the fourth channel - 32 ; The first waterproof apron--103; the second waterproof apron--203.

Detailed ways

In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be further clearly and completely described below in conjunction with the embodiments of the present invention. It should be noted that the described embodiments are only some of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

It should be understood that the orientation or positional relationship indicated by the terms "upper", "lower", "front", "rear", "left", "right" etc. are based on the orientation or positional relationship shown in the drawings, and are only In order to facilitate the description of the present invention and simplify the description, it does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention.

Terms such as "first", "second", "third", "fourth" are used for descriptive purposes only, and should not be understood as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, references to "first", "second", "third" and "fourth" features may expressly or implicitly include one or more of these features.

Example 1

As shown in Figure 1, Figure 2 and Figure 3, a micro-pitch electroporation device with adjustable electrode spacing includes a fixed electrode 1, an adjustable electrode 2, an electrode seat 3, a bolt seat 4, and an adjustment bolt 5;

The fixed electrode 1 is installed on one side of the electrode holder 3, and the bolt holder 4 is installed on the other side of the electrode holder 3; one side of the adjustable electrode 2 is slidably installed in the electrode holder 3, and the One side of the adjustable electrode 2 is in sliding contact with or spaced from one side of the fixed electrode 1; the adjusting bolt 5 is threaded through a threaded hole of the bolt seat 4 and the other side of the adjustable electrode 2 or fixed connection;

The fixed electrode 1 is used to cooperate with the adjustable electrode 2 to apply the voltage required for cell electroporation to the cell fluid in the device;

The adjustable electrode 2 is used to cooperate with the fixed electrode 1 to apply the voltage required for cell electroporation to the cell fluid in the device;

The electrode holder 3 is used to install the fixed electrode 1 and the adjustable electrode 2, so that the cell fluid forms a cell fluid flow channel in the electrode holder 3;

The bolt seat 4 is used to screw the adjusting bolt 5, so that the adjusting bolt 5 is displaced relative to the electrode holder 3 during the rotation process;

The adjusting bolt 5 is used to link the adjustable electrode 2;

The shape of one side of the fixed electrode 1 is a cylinder, and the other side of the fixed electrode 1 is installed on one side of the electrode holder 3; the shape of the adjustable electrode 2 is a cylinder; the fixed electrode One side of the electrode 1 and one side of the adjustable electrode 2 are both embedded in the same through hole of the electrode holder 3;

The end surface of one side of the fixed electrode 1 is provided with a first inverted tapered concave hole 11; the end surface of one side of the adjustable electrode 2 is provided with a first tapered frustum 21; the first inverted cone Shaped concave hole 11 and the first conical truncated body 21 cooperate with each other or are spaced from each other; the fixed electrode 1 is provided with a first channel 12, and a channel opening of the first channel 12 is opened in the first inverted conical concave The hole bottom surface of the hole 11, the other channel port of the first channel 12 is used for inputting or outputting the cell fluid; the electrode base 3 is provided with a second channel 31, and a channel port of the second channel 31 is opened on the electrode The inner wall surface of a through hole of the seat 3 is located on the side of the end face of one side of the fixed electrode 1 fixedly installed on the electrode seat 3, and the other channel port of the second channel 31 is used for inputting or outputting the cell fluid .

Example 2

Embodiment 2 is the further optimization of embodiment 1;

As shown in Fig. 1, Fig. 2 and Fig. 3, the taper of the conical surface of the first inverted tapered concave hole 11 is the same as the taper of the conical surface of the first conical platform body 21;

The adjusting bolt 5 is an M3 bolt; the pitch of the adjusting bolt 5 is 0.5mm; the taper of the conical surface of the first inverted conical concave hole 11 is 1:5; the first conical platform 21 The taper of the conical surface is 1:5.

Example 3

Embodiment 3 is the further optimization of embodiment 1;

As shown in Fig. 1, Fig. 2 and Fig. 3, the fixed electrode 1 is installed on one side of the electrode holder 3 by several screws; the bolt seat 4 is installed on the other side of the electrode holder 3 by several screws.

Example 4

Embodiment 4 is the further optimization of embodiment 1;

As shown in Figure 1, Figure 2 and Figure 3, a first waterproof rubber ring 103 is arranged between the cylindrical side wall of the fixed electrode 1 and the through hole wall of the electrode seat 3; the cylindrical side of the adjustable electrode 2 A second waterproof rubber ring 203 is arranged between the wall and the through hole wall of the electrode holder 3 .

Example 5

As shown in Figure 1, Figure 4, and Figure 5, a micro-pitch electroporation device with adjustable electrode spacing includes a fixed electrode 1, an adjustable electrode 2, an electrode seat 3, a bolt seat 4, and an adjustment bolt 5;

The fixed electrode 1 is installed on one side of the electrode holder 3, and the bolt holder 4 is installed on the other side of the electrode holder 3; one side of the adjustable electrode 2 is slidably installed in the electrode holder 3, and the One side of the adjustable electrode 2 is in sliding contact with or spaced from one side of the fixed electrode 1; the adjusting bolt 5 is threaded through a threaded hole of the bolt seat 4 and the other side of the adjustable electrode 2 or fixed connection;

The fixed electrode 1 is used to cooperate with the adjustable electrode 2 to apply the voltage required for cell electroporation to the cell fluid in the device;

The adjustable electrode 2 is used to cooperate with the fixed electrode 1 to apply the voltage required for cell electroporation to the cell fluid in the device;

The electrode holder 3 is used to install the fixed electrode 1 and the adjustable electrode 2, so that the cell fluid forms a cell fluid flow channel in the electrode holder 3;

The bolt seat 4 is used to screw the adjusting bolt 5, so that the adjusting bolt 5 is displaced relative to the electrode holder 3 during the rotation process;

The adjusting bolt 5 is used to link the adjustable electrode 2;

The shape of one side of the fixed electrode 1 is a cylinder, and the other side of the fixed electrode 1 is installed on one side of the electrode holder 3; the shape of the adjustable electrode 2 is a cylinder; the fixed electrode One side of the electrode 1 and one side of the adjustable electrode 2 are both embedded in the same through hole of the electrode holder 3;

The end surface of one side of the fixed electrode 1 is provided with a second tapered frustum 13; the end surface of one side of the adjustable electrode 2 is provided with a second inverted tapered concave hole 22; the second inverted cone Shaped concave hole 22 and the second conical frustum 13 are mutually matched or mutually spaced; the fixed electrode 1 is provided with a third passage 14, and a channel opening of the third passage 14 is opened in the second conical frustum 13, another channel port of the third channel 14 is used to input or output cell fluid; the electrode base 3 is provided with a fourth channel 32, and a channel port of the fourth channel 32 is opened on the electrode The inner wall surface of a through hole of the seat 3 is located on the side of the end face of one side of the fixed electrode 1 fixedly installed on the electrode seat 3, and the other channel port of the fourth channel 32 is used for inputting or outputting the cell fluid .

Example 6

Embodiment 6 is the further optimization of embodiment 5;

As shown in FIG. 1 , FIG. 4 , and FIG. 5 , the taper of the tapered surface of the second inverted tapered concave hole 22 is the same as the taper of the tapered surface of the second tapered platform body 13 .

Example 7

Embodiment 7 is the further optimization of embodiment 5;

As shown in Fig. 1, Fig. 4, Fig. 5, described adjustment bolt 5 is M3 bolt; The pitch of described adjustment bolt 5 is 0.5mm; :5; the taper of the conical surface of the second conical frustum 13 is 1:5.

Example 8

Embodiment 8 is the further optimization of embodiment 5;

As shown in Fig. 1, Fig. 4 and Fig. 5, the fixed electrode 1 is installed on one side of the electrode holder 3 by several screws; the bolt seat 4 is installed on the other side of the electrode holder 3 by several screws.

Example 9

Embodiment 9 is the further optimization of embodiment 5;

As shown in Fig. 1, Fig. 4 and Fig. 5, a first waterproof rubber ring 103 is arranged between the cylindrical side wall of the fixed electrode 1 and the through hole wall of the electrode seat 3; the cylindrical side of the adjustable electrode 2 A second waterproof rubber ring 203 is arranged between the wall and the through hole wall of the electrode holder 3 .

从而实现电极间距的微细调节，通过在电极座和可调电极上标示相应刻度(类似游标卡尺)，即可实现电极间距的精细调节，调节精度可超过1μm。The working principle of the present invention is: the fixed electrode and the adjustable electrode are made into a pair of conical surfaces paired with each other, and the distance between the two conical surfaces is the actual electrode spacing in the electroporation process; the fixed electrode is fixed on the electrode seat On the other hand, the other end of the adjustable electrode is connected to the adjusting bolt through the thread, and the adjusting bolt is fixed on the bolt seat. By rotating the adjustable electrode, the left and right movement of the adjustable electrode can be realized; in the embodiment, an M3 with a pitch of 0.5mm is used. Screw, the taper of the two electrode tapered surfaces is designed to be 1:5, if the adjustable electrode rotates one circle, it will move 0.5mm in the horizontal direction, while the distance between the two tapered surfaces changes

In this way, the fine adjustment of the electrode spacing can be realized. By marking the corresponding scale (similar to a vernier caliper) on the electrode holder and the adjustable electrode, the fine adjustment of the electrode spacing can be realized, and the adjustment accuracy can exceed 1 μm.

The working process of the present invention: the cell liquid enters the cell liquid cavity of the electroporation device from the channel of the electrode base (the size of the cell liquid cavity is determined by the gap distance between the fixed electrode and the adjustable electrode), and at this time, a corresponding voltage is applied to the electrode , the cell fluid is electroporated, and the cell fluid continues to pass through the channel in the fixed electrode to reach the outlet and flow out of the electroporation device. With the continuous flow of the cell fluid, the voltage is continuously applied to the electrode, thereby realizing the continuous flow operation of cell electroporation.

The above-mentioned embodiments only express several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but should not be construed as limiting the patent scope of the present invention. It should be pointed out that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

Claims (6)

1. A micro-space electroporation device with adjustable electrode space is characterized by comprising a fixed electrode (1), an adjustable electrode (2), an electrode seat (3), a bolt seat (4) and an adjusting bolt (5);

the fixed electrode (1) is arranged on one side of the electrode seat (3), and the bolt seat (4) is arranged on the other side of the electrode seat (3); one side part of the adjustable electrode (2) is slidably arranged in the electrode seat (3), and one side part of the adjustable electrode (2) is in sliding contact with or spaced from one side part of the fixed electrode (1); the adjusting bolt (5) passes through a threaded hole of the bolt seat (4) and is in threaded or fixed connection with the other side part of the adjustable electrode (2);

the fixed electrode (1) is used for being matched with the adjustable electrode (2) so as to apply voltage required by cell electroporation to cell fluid in the device;

the adjustable electrode (2) is used for being matched with the fixed electrode (1) so as to apply voltage required by cell electroporation to cell fluid in the device;

the electrode holder (3) is used for mounting the fixed electrode (1) and the adjustable electrode (2), so that cell sap forms a cell sap flow channel in the electrode holder (3);

the bolt seat (4) is used for screwing an adjusting bolt (5), so that the adjusting bolt (5) is displaced relative to the electrode seat (3) in the rotating process;

the adjusting bolt (5) is used for linking the adjustable electrode (2);

one side part of the fixed electrode (1) is in a cylindrical shape, and the other side part of the fixed electrode (1) is arranged on one side of the electrode seat (3); the shape of the adjustable electrode (2) is a cylinder; one side part of the fixed electrode (1) and one side part of the adjustable electrode (2) are embedded into the same through hole of the electrode holder (3); a first inverted conical concave hole (11) is formed in the end face of one side portion of the fixed electrode (1); a first conical table body (21) is arranged on the end face of one side part of the adjustable electrode (2); the first inverted conical concave hole (11) and the first conical table body (21) are mutually matched or mutually spaced; the fixed electrode (1) is provided with a first channel (12), one channel opening of the first channel (12) is arranged on the bottom surface of the first inverted conical concave hole (11), and the other channel opening of the first channel (12) is used for inputting or outputting cell sap; the electrode holder (3) is provided with a second channel (31), one channel port of the second channel (31) is arranged on the inner wall surface of one through hole of the electrode holder (3) and is positioned beside the end surface of one side part of the fixed electrode (1) fixedly arranged on the electrode holder (3), and the other channel port of the second channel (31) is used for inputting or outputting cell sap;

the taper of the conical surface of the first inverted conical concave hole (11) is the same as that of the conical surface of the first conical table body (21);

the adjusting bolt (5) is an M3 bolt; the thread pitch of the adjusting bolt (5) is 0.5mm; the taper of the conical surface of the first inverted conical concave hole (11) is 1; the taper of the tapered surface of the first tapered frustum (21) is 1.

2. The fine pitch electroporation apparatus with adjustable electrode pitch according to claim 1, wherein a second tapered frustum (13) is provided on an end surface of one side portion of the fixed electrode (1); a second inverted conical concave hole (22) is formed in the end face of one side portion of the adjustable electrode (2); the second reverse tapered concave hole (22) and the second tapered platform body (13) are mutually matched or mutually spaced; the fixed electrode (1) is provided with a third channel (14), one channel opening of the third channel (14) is arranged on the table top surface of the second conical table body (13), and the other channel opening of the third channel (14) is used for inputting or outputting cell sap; the electrode holder (3) is provided with a fourth channel (32), one channel opening of the fourth channel (32) is arranged on the inner wall surface of one through hole of the electrode holder (3) and is positioned beside the end surface of one side part of the fixed electrode (1) fixedly arranged on the electrode holder (3), and the other channel opening of the fourth channel (32) is used for inputting or outputting cell sap.

3. The micro-interval electroporation device with adjustable inter-electrode distance according to claim 2, wherein the taper of the tapered surface of the second inverted conical concave hole (22) is the same as the taper of the tapered surface of the second conical frustum (13).

4. The fine pitch electroporation apparatus with adjustable electrode pitch according to claim 3, wherein the adjusting bolt (5) is an M3 bolt; the thread pitch of the adjusting bolt (5) is 0.5mm; the taper of the tapered surface of the second inverted tapered concave hole (22) is 1; the taper of the conical surface of the second conical frustum (13) is 1.

5. The fine pitch electroporation device with adjustable electrode spacing according to claim 1, wherein the fixed electrode (1) is mounted on one side of the electrode holder (3) by a plurality of screws; the bolt seat (4) is arranged on the other side of the electrode seat (3) through a plurality of screws.

6. The micro-interval electroporation device with adjustable electrode interval according to claim 1, wherein a first waterproof rubber ring (103) is arranged between the cylindrical side wall of the fixed electrode (1) and the through hole wall of the electrode holder (3); and a second waterproof rubber ring (203) is annularly arranged between the cylindrical side wall of the adjustable electrode (2) and the through hole wall of the electrode seat (3).

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