CN210151124U - Pressurized flow type electrotransfection device - Google Patents

Abstract

The utility model discloses a can pressor STREAMING electrotransfection device provides one kind and realizes automatic feed liquor and pressurization through mobilizable scalable chock plug, restraines the production of bubble in the electrotransfection in-process cell liquid to effectively clear away the electrotransfection device that accumulates pastes such as dead cell on attached to electrode and body inner wall.

Description

Pressurized flow type electrotransfection device

Technical Field

The utility model relates to a can pressor STREAMING electrotransfection device, more specifically, the utility model relates to a carry out pressor mode through portable piston and restrain the STREAMING electrotransfection device that the bubble produced and clear away the paste that accumulates on the electrode among the electrotransfection process.

Background

The cell membrane is a thin membrane surrounding the cell periphery and is a permeable barrier for selective exchange of substances between the cell and the outside. The cell membrane makes the cell an independent life unit and has a relatively stable internal environment. Some substances in the surrounding environment may pass through the cell membrane, others do not. Cells can take up nutrients from the surrounding environment through the cell membrane, excrete metabolites, and allow the transport of substances to reach an equilibrium state. Therefore, the basic function of the cell membrane is to maintain a relatively stable intracellular microenvironment and selectively exchange substances with the external environment.

It is found that if a certain intensity of electric stimulation is applied to cells for a certain period of time, micropores can be induced on cell membranes, so that the permeability of the cells is enhanced, and the cell Electroporation (Electroporation) refers to a biophysical process of the cells under the action of an applied pulse electric field, wherein transient micropores are formed on cell membrane lipid bilayers. Electrotransfection (Electrotransfection) is a technique for introducing foreign biological macromolecules, such as DNA, RNA or proteins, into cells using electroporation. When the cell membrane is subjected to electroporation, the permeability and the membrane conductance of the cell membrane are increased instantaneously, so that molecules, such as hydrophilic molecules, DNA, proteins, virus particles, drug particles and the like, which cannot pass through the cell membrane under normal conditions can enter the cell through micropores. After the electrical stimulation is removed in a short time, the micropores in the cell membrane disappear, and the cell membrane becomes a selective permeability barrier again.

Compared with the traditional chemical transfection and virus transfection, the electrotransfection has the advantages of no chemical pollution, no permanent damage to cells, high efficiency and the like, and has wide application prospect in the fields of biophysics, molecular biology, clinical medicine and the like.

Although the mechanism of electrotransfection is not completely understood, it is well known in this context that cell electrotransfection involves the movement of the lipid bilayer of the cell membrane, resulting in the formation of transient micropores in the membrane, allowing exogenous molecules to enter the cell through the micropores.

In the prior art, there are three main types of methods for completing the process of electrotransfection of cells: the cells are placed between a pair of parallel electrodes spaced a few millimeters to a few centimeters apart. The cells are electrically stimulated in an electric field between the electrodes for the purpose of electrotransfection. For example, US patent No. 5389069.

The micro needle electrode is pricked into tissue or cell fluid to shock the cell electrically to reach the aim of electrotransfection. For example, US patent No. 5389069.

A chamber is placed between a pair of parallel electrodes so that the cell suspension is shocked while flowing in the chamber. For example, US patent US 6773669.

Chinese patent CN201010242144 discloses a flow electrotransfection device and system, the system includes: a flow electrotransfection device comprising: the electrode is arranged in parallel and in pairs, and each pair of electrodes comprises an anode and a cathode which are oppositely arranged; a channel disposed over the electrode that restricts fluid flow; the starting end of the channel is provided with a plurality of inlet branch channels which are converged into a main channel, the ending end of the channel is provided with a plurality of outlet branch channels, and a top cover with a plurality of fluid inlets and outlets is arranged above the channel; the injection pump is connected to the inlet and the outlet of the top cover in the flow type electrotransfection device through pipelines to control the flow rate of the fluid; and the voltage source is connected with the electrode set by the electric connector and generates pulse voltage. The flow-type electrotransfection system utilizes the fluid channel and the connected injection pump to realize the continuous flow of various cell suspensions in the fluid channel, thereby enabling the process that the cells are electrotransfected to be continuously carried out and realizing the rapid processing of a large number of samples.

Chinese patent CN201610806987 discloses a disposable article for the electrotransfection of cells comprising: a fluid compartment inside the disposable; a first fluid port for providing a cell suspension to the fluid compartment; and a second fluid port for delivering a fluid comprising at least one compound to be electrotransfected into the cell to the fluid compartment; a first electrode and a second electrode disposed in the fluid compartment; at least one outlet port delivering the fluid from the fluid compartment, wherein the first and second fluid ports are in fluid communication with a mixing channel in fluid communication with the fluid compartment.

However, the flow electrotransfection device disclosed above generates a large amount of bubbles during the actual electrotransfection process, and the generated bubbles adhere to the electrode plates to affect the uniformity of the electric field, thereby causing the electrotransfection effect to be unstable. The inventors have made some efforts to reduce the generation of air bubbles during electrotransfection by pressurizing the suspension of the electrotransfected cells, for example CN 20181049356 discloses an intermittent flow electrotransfection device; CN2018110783561 discloses an intermittent flow type electrotransfection device; CN2018109939121 discloses a flow-type electrotransfection device; although the above patent has achieved certain technical effects, the above patent does not affect the present invention.

Meanwhile, the inventor also finds that paste is remained on the inner wall of the tube body and the electrode in the electrotransfection process, so that the electric field intensity and the uniformity of the electric field are influenced.

Disclosure of Invention

The utility model provides a to the technical problem that above-mentioned prior art exists, provide one kind through the automatic feed liquor of mobilizable piston and pressurized mode, restrain the production of bubble in the electrotransfection in-process cell liquid to effectively detach the electrotransfection device of remaining the paste on body inner wall and electrode.

Another object of the present invention is to improve the stability of the electrotransfection device and to improve the electrotransfection efficiency.

In order to achieve the above objects, the technical scheme of the electrotransfection device provided by the present invention is summarized as follows:

an electrotransfection device comprises a tube body, a first electrode, a second electrode and a piston, wherein the first electrode and the second electrode are arranged in parallel on the inner wall of the tube body, one end of the tube body is provided with a sealing end, the other end of the tube body is provided with the piston capable of moving along the tube body relatively, the piston comprises a telescopic plug head and a connecting rod, a cavity for containing a liquid sample is arranged inside the telescopic plug head and the tube body, the tube body is provided with at least one liquid inlet and/or outlet and at least one liquid inlet and/or outlet valve, the liquid inlet and/or outlet is positioned on the sealing end of the tube body, and the liquid inlet and/or outlet valve is positioned on the liquid inlet and/or outlet of the tube body.

Preferably, the cross-sectional area of the retractable stopper is equal to the cross-sectional area of the cavity. When the telescopic plug head is contracted, the sectional area of the telescopic plug head is smaller than that of the cavity.

Preferably, the material of the retractable plug head of the piston is an electroactive polymer, and the electroactive polymer can deform under the electric stimulation. The electroactive polymer is preferably polyvinylidene fluoride, collagen fibers, ion-exchange polymeric metals, or the like. In the case of power-on, the electroactive polymer contracts, and in the case of power-off, the electroactive polymer expands.

Preferably, the connecting rod of the piston is hollow, and a lead is arranged in the connecting rod and connected with the electroactive polymer to transmit an electric signal and deform the electroactive polymer, so that the expansion effect of the telescopic plug head is achieved.

Preferably, the length of the first electrode and the second electrode is less than the length of the tube, and one end of the first electrode and the second electrode is flush with the sealed end of the tube.

Preferably, the piston is provided with a working position, and the working position is the position where the first electrode and the second electrode are far away from the other ends of the sealed ends of the tube body, namely the height of the liquid sample to be processed in the cavity is flush with the upper ends of the first electrode and the second electrode.

Preferably, the retractable stopper and the interior of the body form a closed chamber when the liquid inlet and/or outlet valve is closed.

Preferably, the device further comprises a push-pull device enabling the piston to move relatively along the tube, the push-pull device being connected with the connecting rod; the push-pull device includes, but is not limited to, a hydraulic rod, a pneumatic rod, a spiral push-pull device, an electromagnetic push-pull device, a manual push-pull device, etc.

Preferably, the device further comprises a conduit in sealed connection with the liquid inlet and/or outlet.

The utility model discloses beneficial effect that technical scheme brought:

the electrotransfection device of the utility model can continuously process a large amount of liquid samples, and can effectively remove residual paste on the inner wall of the tube body and the electrode, thereby improving the electrotransfection efficiency;

the utility model discloses an electrotransfection device is to liquid pressurization at the electrotransfection in-process, and the suppression bubble produces, reduces because of the attached electric field intensity inhomogeneity that leads to of bubble, improves electrotransfection device's stability to improve electrotransfection efficiency.

The utility model discloses an electrotransfection device structural design is novel, can automatic feed liquor, has pioneering nature.

Drawings

FIG. 1 is an illustration showing an overall structure of an electrotransfection device of the present invention;

FIG. 2 is a schematic view of the electrotransfection device of the present invention with liquid entering the chamber;

FIG. 3 is a schematic view of the electrotransfection device of the present invention, wherein the electrotransfection device is pressurized to perform electrotransfection after the liquid completely enters the cavity;

FIG. 4 is a schematic view of the electrotransfection device of the present invention removing the liquid after completion of electrotransfection;

the attached drawings are annotated: 1-valve, 2-tube sealing end, 3-wire, 4-telescopic plug, 5-connecting rod, 6-first electrode, 7-second electrode, 8-cavity and 9-tube.

Detailed Description

Example one

Overall structure design 1

The electrotransfection device provided by the utility model is shown in figure 1 and comprises a tube body (9), wherein one end of the tube body (9) is open, and the other end is provided with a tube body sealing end (2); the device further comprises a first electrode (6) and a second electrode (7), wherein the length of the first electrode (6) and the length of the second electrode (7) are smaller than that of the tube body (9), and one end of the first electrode (6) and one end of the second electrode (7) are flush with the sealed end (2) of the tube body; the sealing end (2) of the pipe body is provided with a valve (1) for controlling the liquid to flow in and out; the device further comprises a telescopic plug head (4) and a connecting rod (5), wherein a cavity (8) for containing a liquid sample is formed inside the telescopic plug head (4) and the tube body (9), and polyvinylidene fluoride is selected as a material of the telescopic plug head (4); a lead (3) is embedded in the connecting rod (5); the lead (3) is electrically connected with the telescopic plug head (4); scalable chock plug (4) can reciprocate in connecting rod (5), when wire (3) outage, scalable chock plug (4) inflation can make cavity (8) form sealed state. When the conducting wire (3) is electrified, the telescopic plug head (4) contracts.

Example two

Method for using device

The electrotransfection device provided by the utility model is used in the way as shown in figures 2-4, and firstly, a sample is injected. The valve (1) is opened, and the connecting rod (5) is pushed to enable the telescopic plug head (4) to be located at the sealing end (2) of the pipe body; adding a liquid sample to be treated above the telescopic plug head (4) through a liquid-transferring gun; the valve (1) is closed, the lead (3) is electrified, the telescopic plug head (4) is contracted, the connecting rod (5) is pulled to enable the telescopic plug head (4) to move to the working position far away from the sealing end (2) of the tube body, the air pressure in the cavity (8) is smaller than the external air pressure at the moment, and as shown in figure 2, a liquid sample to be processed added above the telescopic plug head (4) enters the cavity (8) from a gap between the telescopic plug head (4) and the tube wall under the influence of the air pressure.

When the height of the liquid sample to be processed in the cavity (8) is flush with the upper ends of the first electrode (6) and the second electrode (7), namely the working position of the piston, the conducting wire (3) is powered off, so that the telescopic plug head (4) is expanded, the section of the telescopic plug head (4) is the same as the section area of the cavity (8), and as shown in figure 3, the cavity (8) filled with liquid is pressurized by pushing the connecting rod (5).

And (3) performing electrotransfection while pressurizing, after the electrotransfection is finished, opening the valve (1) and pushing the connecting rod (5) to enable the telescopic plug head (4) to be arranged at the sealing end (2) of the tube body, and discharging the processed liquid sample out of the cavity (8) as shown in figure 4. The above operation was repeated to process the sample again.

Although the present invention has been described in detail, modifications within the spirit and scope of the invention will be apparent to those skilled in the art. It should be understood that aspects of the invention and portions of the various embodiments and various features described above and/or in the appended claims may be combined or interchanged either in whole or in part. As will be appreciated by one skilled in the art, in the foregoing description of the various embodiments, those embodiments referring to another embodiment may be combined with other embodiments as appropriate. Furthermore, those of ordinary skill in the art will appreciate that the foregoing description is by way of example only, and is not intended to limit the invention.

Claims (9)

1. An electrotransfection device comprises a tube body, a first electrode, a second electrode and a piston, wherein the first electrode and the second electrode are arranged in parallel on the inner wall of the tube body, one end of the tube body is provided with a sealing end, the other end of the tube body is provided with the piston capable of moving along the tube body relatively, the piston comprises a telescopic plug head and a connecting rod, a cavity for containing a liquid sample is arranged inside the telescopic plug head and the tube body, the tube body is provided with at least one liquid inlet and/or outlet and at least one liquid inlet and/or outlet valve, the liquid inlet and/or outlet is positioned on the sealing end of the tube body, and the liquid inlet and/or outlet valve is positioned on the liquid inlet and/or outlet of the tube body.

2. The electrotransfection device of claim 1, wherein the material of the retractable stopper is an electroactive polymer.

3. The electrotransfection device according to claim 2, wherein the electroactive polymer is polyvinylidene fluoride, collagen fibers, ion-exchange polymeric metal.

4. The electrotransfection device of claim 1, wherein the cross-sectional area of the retractable stopper is equal to the cross-sectional area of the chamber.

5. The electrotransfection device according to claim 4, wherein the retractable stopper and the interior of the tube form a closed chamber when the liquid inlet and/or outlet valve is closed.

6. The electrotransfection device of claim 1, wherein the cross-sectional area of the retractable stopper is less than the cross-sectional area of the chamber when the retractable stopper is retracted.

7. The electrotransfection device according to claim 1, wherein a wire is disposed inside the linkage.

8. The electrotransfection device according to claim 7, wherein the wire is electrically connected to an electroactive polymer.

9. The electrotransfection device of claim 1, wherein the first and second electrodes have a length less than the length of the tube.

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