CN117431158A

CN117431158A - Flow electroporation device

Info

Publication number: CN117431158A
Application number: CN202210829909.2A
Authority: CN
Inventors: 戴晓兵
Original assignee: Etta Biotech Co Ltd
Current assignee: Etta Biotech Co Ltd
Priority date: 2022-07-14
Filing date: 2022-07-14
Publication date: 2024-01-23
Also published as: WO2024012261A1

Abstract

The invention discloses a flow electroporation device which comprises a host, an electrode assembly arranged on the host, a sample injection container and a collection container. The electrode assembly comprises an electrode for electroporation treatment of the cell suspension, the electrode assembly is provided with a liquid inlet and a liquid outlet, and the electrode assembly can be arranged on one side of the host in a relatively rotatable manner; the sample injection container is used for storing cell suspension to be treated by the electrode and is communicated with the liquid inlet; the collecting container is used for receiving the cell suspension treated by the electrode, and the collecting container is communicated with the liquid outlet. The flow electroporation device has a first working state and a second working state, and in the first working state, the electrode performs electroporation treatment on the cell suspension; the electrode assembly rotates relative to the host to be converted into a second working state; in the second operating state, at least part of the cell suspension in the electrode assembly can flow out of the liquid outlet under the action of gravity. The invention can reduce bubble generation during cell treatment, improve cell treatment efficiency and improve cell recovery rate after treatment.

Description

Flow electroporation device

Technical Field

The invention relates to the technical field of biology, in particular to a flow electroporation device.

Background

Electroporation (electric corporation) is a technique that uses an electric field to act on a cell membrane to make the cell membrane generate micropores through which foreign molecules such as DNA, RNA, proteins pass, and the like, which are instantaneously generated by the electric field and can be recovered as the electric field disappears, thereby avoiding permanent damage to cells. The electroporation technology can be used for introducing various exogenous molecules such as DNA, RNA, protein, sugar, dye, virus particles and the like into prokaryotic or eukaryotic cells, is called an Electrotransfection technology (Electrotransfection), has wide application in the technical fields of antibody protein production, in Vitro Diagnosis (IVD) reagent raw material production, immune cell gene editing, gene modification and the like, and is an important technology for delivering substances into cells.

There are various methods for electroporation of cells, among which, the flow electroporation technique can continuously electroporate a flowing cell-containing suspension (hereinafter referred to as a cell suspension), which is highly efficient and particularly suitable for batch processing of large-volume cell suspensions. A common flow electroporation device mainly comprises a pair of planar electrodes for generating an electric field, and a chamber between the two planar electrodes, through which a cell suspension can be electrically shocked while flowing, and electroporation is generated, so that exogenous molecules in the cell suspension can enter target cells. In the prior art, the flow electroporation device mostly has the problems of low cell treatment efficiency, large cell suspension residual quantity and the like. On one hand, when the cell suspension receives electric shock, a water electrolysis phenomenon can be generated, a large number of bubbles are easy to generate, the problems of discontinuous or uneven liquid flow, unstable flow speed and the like are caused, the uniformity of an electric field is reduced, and meanwhile, the controllability of the times of receiving the electric shock by the cells is influenced, so that the transfection efficiency of the cells is reduced; on the other hand, bubbles generated during electroporation can form foam, which can mix with cells to easily cause cell suspension to remain in the chamber and tubing, which is difficult to drain, reducing the final cell recovery rate of the flow electroporation device.

Disclosure of Invention

The invention aims to solve the problems in the prior art and provides a flow electroporation device with high cell treatment efficiency.

In order to achieve the above purpose, the invention adopts the following technical scheme:

the flow electroporation device comprises a host, an electrode assembly, a sample injection container and a collection container, wherein the electrode assembly, the sample injection container and the collection container are respectively arranged on the host,

the electrode assembly comprises an electrode for electroporation treatment of the cell suspension, the electrode assembly is provided with a liquid inlet and a liquid outlet, and the electrode assembly can be arranged on one side of the host in a relatively rotatable manner;

the sample injection container is used for storing cell suspension to be treated by the electrode, and is communicated with the liquid inlet through a liquid inlet pipeline;

the collecting container is used for receiving the cell suspension treated by the electrode, and is communicated with the liquid outlet through a liquid outlet pipeline;

the flow electroporation device has a first working state and a second working state, and when the flow electroporation device is in the first working state, the electrode performs electroporation treatment on the cell suspension; the electrode assembly rotates relative to the host during the transition of the flow electroporation device from the first operating state to the second operating state; when the flow electroporation device is in the second operating state, at least a portion of the cell suspension in the electrode assembly is capable of flowing out of the liquid outlet under the force of gravity.

In some embodiments, when the flow electroporation device is in the first operating state, the height of the liquid inlet is lower than the height of the liquid outlet; when the flow electroporation device is in the second working state, the height of the liquid outlet is reduced. When the flow electroporation device is in the first working state, the flow electroporation device can normally run, the flow direction of the cell suspension flowing in and out from the lower part continuously flows through the electrode assembly and electroporation treatment is carried out, the cavity between the electrode plates can be filled under the action of the gravity of the cell suspension, the electric communication between the electrode plates is kept, the continuity and uniformity of liquid flow are improved, the generation of bubbles is effectively reduced, the uniformity of an electric field is improved, and thus the electroporation efficiency is comprehensively improved. When the cell suspension is in the second working state, the height of the liquid outlet is reduced, so that the cell suspension which is not discharged in the cavity can be discharged under the action of self gravity.

In some embodiments, when the flow electroporation device is in the second operating state, the height of the liquid outlet is lower than the height of the liquid inlet. So that the electrode assembly has a larger inclination angle, which contributes to more sufficient discharge of the cell suspension.

In some embodiments, when the flow electroporation device is in the first working state, the axis of the liquid inlet and the axis of the liquid outlet extend along a vertical direction. Therefore, in the first working state, the cell suspension vertically upwards flows through the electrode assembly, and the uniformity of liquid flow and the uniformity of an electric field can be better ensured.

In some embodiments, the electrode comprises a first electrode and a second electrode which are arranged at intervals, a treatment channel is arranged between the first electrode and the second electrode, the flow electroporation device is in the first working state, the treatment channel extends along the up-down direction, and the cell suspension flows through the treatment channel from bottom to top. Therefore, the cell suspension not only enters and exits from the liquid inlet and the liquid outlet of the whole electrode assembly, but also can keep the flow direction of the lower inlet and the upper outlet in the specific treatment channel between the electrodes, and further ensures the uniformity of liquid flow and electric field uniformity in the chamber between the electrodes.

In some embodiments, the flow electroporation device further comprises a liquid level sensor arranged between the sample injection container and the electrode assembly, the liquid level sensor is fixedly arranged on one side of the host, the height of the liquid level sensor is lower than that of the electrode assembly, and the liquid level sensor is provided with a pipeline guide groove which can be tightly attached to a liquid inlet pipeline, so that the liquid level sensor can work normally. The pipeline guide groove extends along the vertical direction, and the liquid inlet pipeline passes through the pipeline guide groove from bottom to top. The liquid level sensor can be used for measuring the liquid level state in the liquid inlet pipeline and transmitting signals to a control system, and the control system controls the working state of the electrode assembly. The liquid level sensor adopts a working mode of vertically placing and feeding and discharging cell suspension in the sample feeding pipeline, so that liquid is filled in the liquid feeding pipeline in the pipeline guide groove, and misjudgment of detection results caused by liquid in the pipeline or bubbles is avoided, and the control system is influenced to accurately control the electroporation process.

In some embodiments, the flow electroporation device further comprises a peristaltic pump, wherein the peristaltic pump is fixedly arranged on one side of the host, and the liquid outlet pipeline passes through the peristaltic pump from top to bottom. The peristaltic pump is used for providing the power for the cell suspension to flow in the whole flow electroporation device, is arranged between the collecting container and the electrode assembly, is arranged at the rear end and below the electrode assembly, and is combined with a liquid outlet pipeline of the electrode assembly more smoothly. When the peristaltic pump is started, negative pressure is formed in the electrode assembly and the pipeline at the front side of the electrode assembly, and cell suspension in the sample injection container is sucked and flows out; when the whole pipeline is full of liquid, the pressure in the pipeline reaches balance, and the peristaltic pump stably transmits the cell suspension. Compared with the peristaltic pump which is arranged on the liquid inlet pipeline and forms positive pressure in the electrode assembly and the pipeline at the rear side of the electrode assembly to push the cell suspension to flow, the flow velocity of the cell suspension in the invention is more uniform and stable, and better electroporation treatment effect is facilitated.

In some embodiments, the flow electroporation device further comprises a liquid level sensor and a peristaltic pump, the host has a first side wall, and a second side wall and a third side wall which are respectively arranged at two sides of the first side wall with different width directions, the extending direction of the first side wall is intersected with the extending direction of the second side wall, and the extending direction of the first side wall is intersected with the extending direction of the third side wall; the electrode assembly can be arranged on the first side wall in a relatively rotatable manner, and the liquid level sensor and the peristaltic pump are arranged on the first side wall; the sample injection container is arranged on the second side wall, and the collecting container is arranged on the third side wall.

In some embodiments, the first sidewall is perpendicular to the second sidewall, and the first sidewall is also perpendicular to the third sidewall. The spatial layout of the flow electroporation device is optimized, the liquid level sensor, the electrode assembly and the peristaltic pump are arranged on the same side wall, the pipeline arrangement is more orderly, the rotation of the electrode assembly is not easy to cause the crossed winding of the pipeline, and the smooth flow of liquid is ensured; the sample injection container and the collection container are respectively arranged on the left side wall and the right side wall (namely the second side wall and the third side wall), the positions are relatively fixed, the sample injection container and the collection container are not easy to be influenced by the rotation of the electrode assembly, the width of the host is reduced, and the occupied space of the host is reduced.

In some embodiments, the flow electroporation device further comprises a rotating base and a rotation driving device, wherein the rotating base can be connected with the host in a relatively rotating manner around a rotation center line, the rotation driving device is used for driving the rotating base to rotate, and the electrode assembly is detachably arranged on the rotating base and can rotate synchronously with the rotating base. The rotating seat is used as an intermediate part, which is favorable for improving the stability and reliability of the electrode assembly during rotation, and the electrode assembly and the rotating seat synchronously move.

In some embodiments, the rotation center line extends along a horizontal direction, a base line exists, the base line is a ray which is perpendicular to the rotation center line and extends vertically upwards, an end point of the base line is located on the rotation center line, the electrode assembly has a first limit position, a second limit position and a reference position in the process of rotating around the rotation center line, and the rotation direction of the electrode assembly from the reference position to the second limit position is a positive direction; when the electrode assembly is positioned at the reference position, the axial lead of the liquid outlet coincides with the base line; when the electrode assembly is in the first limit position, the included angle between the axis line of the liquid outlet and the base line is-10 degrees; when the electrode assembly is in the second limit position, an included angle between the axis line of the liquid outlet and the base line is 190 degrees.

In some embodiments, a first limiting structure is disposed between the rotating base and the host, and the first limiting structure is used for limiting the electrode assembly to rotate between the first limiting position and the second limiting position. The first limiting structure limits the rotation angle of the electrode assembly, and the electrode assembly can only rotate between preset limit positions. Generally, in a first operating state, the electrode assembly is in a reference position; in the second working state, the included angle between the axial lead of the liquid outlet and the base line is about 150 degrees, and at the moment, residual cell suspension in a pipeline near the liquid outlet of the electrode assembly can be well discharged. The first limiting structure effectively avoids the problem that the pipeline is pulled or knotted possibly caused by an overlarge rotation angle, and improves the operation reliability of the flow electroporation device.

In some embodiments, the host is provided with a supporting plate extending along the up-down direction, and the supporting plate is provided with a limiting hole; the rotating seat is provided with a connecting part, the rotating center line passes through the connecting part, the connecting part is provided with a convex part protruding outwards along the radial direction of the rotating center line, and the connecting part is inserted into the limiting hole; the first limiting structure comprises the convex part and the limiting hole, and a first boss and a second boss which are arranged at intervals are arranged on the side wall of the limiting hole along the circumferential direction of the rotation center line; when the electrode assembly is at the first limit position, the convex part is abutted against the first boss; when the electrode assembly is in the second limit position, the convex part abuts against the second boss. The first limiting structure specifically adopts a structure of matching a simple convex part with two groups of bosses, and has the advantages of simple design, no space occupation and strong reliability.

In some embodiments, the host has a first side wall, the rotating seat can rotate around the rotation center line relatively and be connected with the first side wall, the rotation center line is perpendicular to the first side wall, the rotating seat includes a rotating table, along the extending direction of the rotation center line, a gap is arranged between the rotating table and the first side wall, and the liquid inlet and the liquid outlet are both located at one side of the rotating table far away from the first side wall. The rotating table provides a supporting plane for the rotation of the electrode assembly, preferably, the rotating table and the first side wall keep a movement gap of about 0.5-0.8 mm, so that the rotating table is not excessively protruded and occupies space, friction between the rotating table and the first side wall can be reduced, and the fluency of rotation is improved.

In some embodiments, the rotating base further includes a connecting base, the connecting base passes through the first side wall along an extending direction of the rotation center line, the rotation driving device is arranged on one side of the first side wall, the rotating table is arranged on the other side of the first side wall, a second limiting structure is arranged between the host and the connecting base, and the second limiting structure is used for limiting the rotating table to move towards the first side wall. The second limiting structure can be used for keeping the gap and preventing the rotating table from moving along the axial direction in the rotating process of the rotating seat.

In some embodiments, the connection seat has an annular convex surface, and the extending direction of the convex surface is perpendicular to the rotation center line; the second limit structure comprises a plurality of limit pieces and convex surfaces, the limit pieces are arranged along the circumferential direction of the connecting seat at intervals, the limit pieces and the rotating table are respectively arranged on the two different sides of the first side wall, and each limit piece is propped against the convex surface. The second limiting structure mainly adopts a mode of pushing the connecting seat from the inner side of the first side wall outwards to prevent the rotating table from moving towards the inner side of the first side wall.

In some embodiments, the connecting seat has a first portion and a second portion connected along an extending direction of the rotation center line, an outer diameter of the first portion is larger than an outer diameter of the second portion, a side of the first portion facing the second portion is convex, each limiting member can be relatively rotatably arranged on the first side wall around a self axis, an outer peripheral surface of each limiting member abuts against an outer peripheral surface of the second portion, and the limiting member is in rolling contact with the second portion. The locating part can be in the circumference to the second portion of connecting seat spacing to the locating part adopts rolling friction's mode and second portion contact, helps reducing the friction that spacing caused, improves and rotates seat pivoted smoothness nature.

In some embodiments, the flow electroporation device further comprises a limiting member connected with the host machine in a relatively rotatable manner around a self axis, and a plurality of limiting members are arranged at intervals; the rotating seat further comprises a connecting seat, and the limiting piece is in rolling contact with the outer peripheral surface of the connecting seat. A plurality of limiting members may be used to provide support and rotational guidance for the rotatable mount.

In some embodiments, the host has a first side wall, the rotating seat can be connected with the first side wall in a relatively rotating manner around the rotation center line, the limiting piece is connected with the first side wall, and the connecting seat passes through the first side wall along the extending direction of the rotation center line. The rotating seat and the limiting piece are connected with the first side wall, and the supporting and guiding functions of the limiting piece are more stable and reliable.

Due to the application of the technical scheme, the flow electroporation device provided by the invention realizes the switching between the first working state and the second working state through the rotation of the electrode assembly relative to the host, so that the direction of the liquid outlet can be changed according to the requirement, and after the cell suspension is subjected to electroporation, the residual part or all of the cell suspension can be discharged out of the electrode assembly only by self gravity, thereby improving the cell recovery rate.

Drawings

In order to more clearly illustrate the technical solution of the present invention, the drawings used in the description of the embodiments will be briefly described.

FIG. 1 is a schematic front view of a flow electroporation device according to an embodiment of the present invention in a first operating state;

FIG. 2 is a schematic front view of the flow electroporation apparatus according to the present embodiment in a second operating state;

FIG. 3 is an exploded view of the electrode assembly, the rotating base and the rotating motor according to the present embodiment;

FIG. 4 is a schematic perspective view illustrating assembly of an electrode assembly, a rotating base and a rotating motor in the present embodiment, where the flow electroporation device is in a first working state;

FIG. 5 is a schematic perspective view illustrating assembly of the electrode assembly, the rotating base and the rotating motor in the present embodiment, wherein the flow electroporation device is in a second working state;

FIG. 6 is a schematic partial perspective view of a flow electroporation apparatus according to the present embodiment;

FIG. 7 is a schematic view showing a partial structural decomposition of the flow electroporation device according to the present embodiment;

fig. 8 is a schematic rear view of the rotating seat and the motor bracket in the present embodiment;

wherein: 100. a host; 110. a first sidewall; 111. a control panel; 120. a second sidewall; 121. a first bracket; 130. a third sidewall; 131. a second bracket; 200. an electrode assembly; 210. a liquid inlet; 220. a liquid outlet; 230. a housing; 310. a sample introduction container; 311. a first passage port; 312. a liquid inlet pipeline; 320. a collection container; 321. a second port; 322. a liquid outlet pipeline; 400. a liquid level sensor; 401. a pipeline guide groove; 500. a peristaltic pump; 600. a rotating seat; 610. a mounting groove; 620. a rotating table; 630. a connecting seat; 631. a first section; 632. a second section; 633. a limiting piece; 640. a rotating seat main body; 641. a connection part; 642. a convex portion; 710. a rotating electric machine; 711. a motor main body; 712. an output shaft; 720. a motor bracket; 721. a support plate; 722. a connecting plate; 723. a limiting hole; 723a, a first boss; 723b, a second boss; 1001. rotating the center line; 1002. a base line; 1003. the axis of the liquid outlet.

Detailed Description

Preferred embodiments of the present invention will be described in detail below with reference to the attached drawings so that the advantages and features of the present invention can be more easily understood by those skilled in the art.

Referring to FIGS. 1 to 8, a flow electroporation apparatus for electroporation of a cell suspension is provided. The flow electroporation device comprises a host 100, an electrode assembly 200, a sample introduction container 310, a collection container 320, a liquid level sensor 400, a peristaltic pump 500, a rotary seat 600, a rotary driving device and the like which are arranged on the host 100.

Referring to fig. 1 and 2, in the present embodiment, the host 100 is substantially rectangular, and the illustrated direction is taken as a reference, the host 100 has a first sidewall 110 located at the front side, and a second sidewall 120 and a third sidewall 130 respectively located at the left and right sides of the first sidewall 110, wherein the extending direction of the first sidewall 110 intersects with the extending direction of the second sidewall 120, and the extending direction of the first sidewall 110 intersects with the extending direction of the third sidewall 130. In practice, the first side wall 110, the second side wall 120, and the third side wall 130 extend along the vertical direction, the first side wall 110 is perpendicular to the second side wall 120 and the third side wall 130, the second side wall 120 is parallel to the third side wall 130, the control panel 111 is disposed at the upper portion of the first side wall 110, and a rotation driving device and a control system are disposed inside the host 100.

In this embodiment, a first bracket 121 is disposed on the second sidewall 120, and the first bracket 121 is used to suspend the sample container 310; the third sidewall 130 is provided with a second bracket 131, the second bracket 131 is used for hanging the collecting container 320, the first bracket 121 is relatively located above the second bracket 131, the cell suspension can be transferred from the sample injection container 310 to the collecting container 320, and the sample injection container 310 and the collecting container 320 are both arranged on the outer sides of the corresponding sidewalls.

In this embodiment, the electrode assembly 200, the liquid level sensor 400 and the peristaltic pump 500 are all disposed outside the first sidewall 110. The electrode assembly 200 includes a housing 230 having an inner cavity, in which electrodes (not shown in the figure) for electroporation treatment of the cell suspension are disposed, and the electrode assembly 200 further includes a liquid inlet 210 and a liquid outlet 220 respectively disposed on the housing 230, wherein the liquid inlet 210 and the liquid outlet 220 are disposed on different sides of the housing 230. The electrode assembly 200 is relatively rotatably disposed at one side of the main body 100, and more particularly, is relatively rotatably disposed on the first sidewall 110 about a rotation center line 1001, and the rotation center line 1001 extends perpendicular to the first sidewall 110 and in a horizontal direction. Further, the electrode comprises a first electrode and a second electrode which are arranged at intervals, a treatment channel for flowing the cell suspension is arranged between the first electrode and the second electrode, and a strong electric field is formed between the first electrode and the second electrode, so that electroporation treatment can be carried out when the cell suspension flows through the treatment channel.

In this embodiment, the sample container 310 is used for storing a cell suspension to be treated by an electrode, and the lower portion of the sample container 310 has a first passage port 311, and the first passage port 311 is in communication with the liquid inlet 210 of the electrode assembly 200 through a liquid inlet pipe 312. The collection container 320 is used for receiving the cell suspension processed by the electrode, and the upper part of the collection container 320 is provided with a second channel port 321, and the second channel port 321 is communicated with the liquid outlet 220 of the electrode assembly 200 through a liquid outlet pipeline 322.

In this embodiment, the liquid level sensor 400 and the peristaltic pump 500 are both fixed on the first sidewall 110, and the heights of the liquid level sensor 400 and the peristaltic pump 500 are both lower than the height of the electrode assembly 200. Wherein, the liquid level sensor 400 is disposed between the sample introduction container 310 and the electrode assembly 200, the liquid level sensor 400 has a pipe guide groove 401, the pipe guide groove 401 extends in a vertical direction, and the liquid inlet pipe 312 passes through the pipe guide groove 401 from bottom to top. The liquid level sensor 400 is further electrically connected to a control system in the host 100, and can measure the liquid level state in the liquid inlet pipe 312 and feed back the signal to the control system, and the control system controls the working state of the electrode assembly 200. Peristaltic pump 500 is disposed between collection container 320 and electrode assembly 200, and fluid line 322 passes through peristaltic pump 500 from top to bottom.

Referring to fig. 3 to 5, the rotary holder 600 is relatively rotatably provided at one side of the main body 100 about a rotation center line 1001, and a rotation driving device is used to drive the rotary holder 600 to rotate, and the electrode assembly 200 is detachably provided on the rotary holder 600 and can be rotated in synchronization with the rotary holder 600.

The flow electroporation device has a first operating state and a second operating state driven by the rotary driving device. Referring to fig. 1 and 4, when the flow electroporation device is in the first working state, the electrodes in the electrode assembly 200 can perform electroporation treatment on the cell suspension, the height of the liquid inlet 210 is lower than that of the liquid outlet 220, the axis of the liquid inlet 210 and the axis of the liquid outlet 220 both extend along the vertical direction, the axes are located in the same vertical plane perpendicular to the first sidewall 110, the treatment channel between the electrodes also extends along the vertical direction, and the cell suspension can flow through the treatment channel from bottom to top and receive electric shock. Thus, the whole treatment channel is ensured to be filled with the cell suspension, the flow velocity of the cell suspension is uniform, and air bubbles generated in the electroporation process can be rapidly discharged through the liquid outlet 220 at the top of the shell 230, so that the electroporation effect is not influenced due to retention in the treatment channel.

Referring to fig. 2 and 5, when the flow electroporation device is in the second working state, the electric shock in the electrode assembly 200 is stopped, and the height of the liquid outlet 220 is reduced, so that the cell suspension in the electrode assembly 200 can flow out from the liquid outlet 220 under the action of gravity, and in this embodiment, the height of the liquid outlet 220 is lower than the height of the liquid inlet 210 in the second working state. Specifically, the liquid outlet 220 is turned clockwise by about 150 ° around the rotation center line 1001 and is then switched to the second operation state as compared with the first operation state. Because the cross-sectional area of the treatment channel is larger than that of the liquid outlet pipeline 322, after electroporation treatment, a part of cell suspension can not be discharged due to the small surface tension of the cell suspension and the dead weight of the cell suspension, so that the cell is lost. At this time, after the rotation of the rotating base 600 and the electrode assembly 200 by the rotation driving device is performed by a certain angle, the liquid outlet 220 is lowered, so that the cell suspension retained in the processing channel is fully discharged and recovered to the collection container 320 under the combined action of the gravity and the suction force of the peristaltic pump 500.

It should be noted that, in this embodiment, when the flow electroporation device is in the first working state and the second working state, the peristaltic pump 500 is always kept working, so that the cell suspension can be continuously and sufficiently sucked out of the electrode assembly 200 in cooperation with the gravity of the cell suspension itself. In other embodiments, peristaltic pump 500 may also be suspended when the flow electroporation device is in the second operating state, wherein the cell suspension is expelled from electrode assembly 200 by gravity alone.

When the treatment of one batch of cell suspension is completed, a new batch of cell suspension treatment may be performed. The electrode assembly 200 is rotated counterclockwise about the rotation center line 1001 by about 150 ° by the rotation driving means and then is changed from the second operation state to the first operation state, so that a new round of electroporation of the cell suspension can be started.

Referring to fig. 6 and 7, in the present embodiment, a rotary motor 710 is specifically adopted in the rotary driving device, a motor bracket 720 is fixedly disposed on the host 100, the motor bracket 720 is specifically fixedly disposed on the inner side of the first side wall 110, the motor bracket 720 includes a supporting plate 721 extending in the vertical direction and a connecting plate 722 extending in the horizontal direction, the upper portion of the supporting plate 721 is fixedly disposed on the rear portion of the connecting plate 722, the front portion of the connecting plate 722 is fixedly disposed on the first side wall 110, and a limiting hole 723 is formed in the supporting plate 721. The rotary electric machine 710 includes a motor main body 711 and an output shaft 712, and the output shaft 712 is provided to the motor main body 711 so as to be rotatable relative to each other about a center line 1001. The motor body 711 is fixedly disposed on one side of the support plate 721, the rotating seat 600 is relatively rotatably disposed on the other side of the support plate 721, specifically, the rotating seat 600 is disposed on a side close to the first side wall 110, the motor body 711 is disposed on a side far from the first side wall 110, the output shaft 712 is disposed in the limiting hole 723 and connected to the connection portion 641 of the rotating seat 600, and the output shaft 712 can drive the entire rotating seat 600 to rotate.

The rotating base 600 mainly includes a rotating base 620, a connecting base 630 and a rotating base main body 640, which are sequentially arranged along the extending direction of a rotating center line 1001, and different parts may be integrally formed or separately manufactured and then fixed together. The rotating base main body 640 is in a cuboid shape, the rotating base 620 and the connecting base 630 are both in a disc shape, the outer diameter of the rotating base 620 is larger than that of the connecting base 630, the connecting base 630 penetrates through the first side wall 110 along the extending direction of the rotating center line 1001, the rotating base 620 is located on the outer side of the first side wall 110, and the rotating base main body 640 is located on the inner side of the first side wall 110. The rotating base 600 is provided with a mounting groove 610, an opening of the mounting groove 610 is positioned on the rotating base 620, and the case 230 of the electrode assembly 200 can be inserted into the mounting groove 610 in a form-fitting manner.

Referring to fig. 3, in order to ensure that the cell suspension can only flow into the electrode assembly 200 in a predetermined direction and to ensure the cell processing effect, the present embodiment provides a third limiting structure between the electrode assembly 200 and the rotating base 600 for limiting the mounting of the electrode assembly 200 to the rotating base 600 in only the predetermined direction.

Referring to fig. 4 and 5, in order to improve the reliability of the device, the connection seat 630 and the electrode assembly 200 in this embodiment cannot rotate 360 ° around the rotation center line 1001, but have a first limit position and a second limit position of rotation. Specifically, the position of the electrode assembly 200 when the flow electroporation device is in the first working state is defined as a reference position, there is a virtual base line 1002, the base line 1002 is a ray extending vertically and upwards perpendicular to the rotation center line 1001, and an end point of the base line 1002 is located on the rotation center line 1001, and the direction in which the electrode assembly 200 rotates from the reference position to the second extreme position is defined as a positive direction (clockwise rotation is shown as a positive direction in the figure). When the electrode assembly 200 is at the reference position, the axis 1003 of the liquid outlet coincides with the base line 1002; when the electrode assembly 200 is at the first limit position, the included angle between the axis 1003 of the liquid outlet and the base line 1002 is-10 degrees, that is, the liquid outlet 220 is inclined slightly to the upper left; when the electrode assembly 200 is in the second extreme position, the angle between the axis 1003 of the liquid outlet and the base line 1002 is 190 °, i.e. the liquid outlet 220 is inclined slightly to the left and the lower.

Referring to fig. 7 and 8, a first limiting structure is disposed between the rotating base 600 and the host 100, and the first limiting structure is used for limiting the electrode assembly 200 to rotate only between the first limit position and the second limit position, i.e. to rotate only forward from the first limit position to the second limit position or rotate reversely from the second limit position to the first limit position. It should be noted that, during the normal operation of the flow electroporation device, the first working state and the second working state are mainly related to the foregoing, the electrode assembly 200 generally does not need to be rotated to the first limit position or the second limit position, and the specific inclination angle of the electrode assembly 200 within the rotatable range is also adjustable.

In this embodiment, the first limiting structure includes a protrusion 642 and a limiting hole 723. The rear part of the rotary seat body 640 has the connection part 641, the rotary center line 1001 passes through the connection part 641, the protrusion 642 is fixedly arranged on the connection part 641 and protrudes outwards along the radial direction of the rotary center line 1001, and the connection part 641 and the protrusion 642 are inserted into the limiting hole 723. The side wall of the limiting hole 723 has a first boss 723a and a second boss 723b arranged at intervals along the circumferential direction of the rotation center line 1001, and a central angle between the first boss 723a and the second boss 723b is about 200 °. When the electrode assembly 200 is in the first limit position, the projection 642 abuts on the first boss 723 a; when the electrode assembly 200 is in the second extreme position, the projection 642 abuts against the second boss 723 b.

Referring to fig. 6, in the present embodiment, along the extending direction of the rotation center line 1001, the rotation table 620 is disposed at intervals from the first side wall 110, and the liquid inlet 210 and the liquid outlet 220 are exposed out of the mounting groove 610 and are located at one side of the rotation table 620 away from the first side wall 110. The gap of about 0.5-0.8 mm can be kept between the rotating table 620 and the first side wall 110, so that direct contact friction between the rotating table and the first side wall is avoided, and rotation is prevented from being influenced. In order to maintain the above gap, a second limiting structure is disposed between the host 100 and the connection seat 630, and the second limiting structure is used to limit the movement of the rotating table 620 toward the first side wall 110. Specifically, the connection seat 630 has a first portion 631 and a second portion 632 that are connected along an extending direction of the rotation center line 1001, and an outer diameter of the first portion 631 is larger than an outer diameter of the second portion 632, so that a side of the first portion 631 facing the second portion 632 has an annular convex surface, and the extending direction of the convex surface is perpendicular to the rotation center line 1001, in this embodiment, an outer circumferential surface of the second portion 632 is specifically a cylindrical surface. The second limiting structure includes a limiting member 633, the limiting member 633 includes a plurality of limiting members 633 disposed along a circumferential direction of the second portion 632, the limiting members 633 are specifically rollers, each limiting member 633 can be relatively rotatably disposed on the first side wall 110 around an axis of the limiting member 633, the axis of the limiting member 633 is parallel to the rotation center line 1001, and the limiting member 633 and the rotation driving device are disposed on the same side of the first side wall 110, i.e. on an inner side of the first side wall 110. One axial side of each limiting member 633 abuts against the annular convex surface, and the outer circumferential surface of each limiting member 633 abuts against the outer circumferential surface of the second portion 632, and the limiting members 633 are in rolling contact with the second portion 632. In this embodiment, the limiting members 633 have six limiting members uniformly spaced along the circumferential direction of the second portion 632, which not only plays a role in limiting the axial movement of the rotating table 620, but also can provide rotational guidance for the limiting members 633, and assist in supporting the rotating seat 600, so that the load of the rotating motor 710 is reduced to some extent.

In this embodiment, in order to avoid unsmooth rotation or noise caused by insufficient torque of the rotating electric machine 710, the device is designed to be lightweight, and the load of the rotating electric machine 710 should not exceed 1kg. Specifically, the rotating base 600 is made of a lightweight material, and has a weight of about 0.4kg, and the rotating motor 710 is actually supported with a lower load due to the weight of a part of the rotating base 600 supported by the above-mentioned stopper 633, so that it is possible to stably operate.

In summary, the flow electroporation device provided in this embodiment can not only significantly improve uniformity and continuity of cell suspension flow in electroporation process and improve electroporation efficiency, but also fully discharge cell suspension after treatment and improve cell recovery rate, and the flow electroporation device is stable and reliable in operation and easy to operate.

The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and to implement the same, but are not intended to limit the scope of the present invention, and all equivalent changes or modifications made according to the spirit of the present invention should be included in the scope of the present invention.

Claims

1. A flow electroporation apparatus, characterized in that: the flow electroporation device comprises a host, an electrode assembly, a sample injection container and a collection container, wherein the electrode assembly, the sample injection container and the collection container are respectively arranged on the host,

2. The flow electroporation device of claim 1, wherein: when the flow electroporation device is in the first working state, the height of the liquid inlet is lower than the height of the liquid outlet; when the flow electroporation device is in the second working state, the height of the liquid outlet is reduced.

3. The flow electroporation device of claim 2, wherein: when the flow electroporation device is in the second working state, the height of the liquid outlet is lower than that of the liquid inlet.

4. The flow electroporation device of claim 2, wherein: when the flow electroporation device is in the first working state, the axial lead of the liquid inlet and the axial lead of the liquid outlet extend along the vertical direction.

5. The flow electroporation device of claim 1, wherein: the electrode comprises a first electrode and a second electrode which are arranged at intervals, a treatment channel is arranged between the first electrode and the second electrode, the flow electroporation device is in the first working state, the treatment channel extends along the up-down direction, and the cell suspension flows through the treatment channel from bottom to top.

6. The flow electroporation device of claim 1, wherein: the flow electroporation device further comprises a liquid level sensor arranged between the sample injection container and the electrode assembly, the liquid level sensor is fixedly arranged on one side of the host, the liquid level sensor is provided with a pipeline guide groove, the pipeline guide groove extends along the vertical direction, and the liquid inlet pipeline penetrates through the pipeline guide groove from bottom to top.

7. The flow electroporation device of claim 1, wherein: the flow electroporation device further comprises a peristaltic pump, the peristaltic pump is fixedly arranged on one side of the host, and the liquid outlet pipeline passes through the peristaltic pump from top to bottom.

8. The flow electroporation device of claim 1, wherein: the flow electroporation device further comprises a liquid level sensor and a peristaltic pump, the host is provided with a first side wall, a second side wall and a third side wall, the second side wall and the third side wall are respectively arranged on two sides of the first side wall, which are different in width direction, the extending direction of the first side wall is intersected with the extending direction of the second side wall, and the extending direction of the first side wall is intersected with the extending direction of the third side wall;

the electrode assembly is connected with the first side wall in a relatively rotatable manner, and the liquid level sensor and the peristaltic pump are arranged on the first side wall; the sample injection container is arranged on the second side wall, and the collecting container is arranged on the third side wall.

9. The flow electroporation device of claim 1, wherein: the flow electroporation device further comprises a rotating seat and a rotary driving device, wherein the rotating seat can relatively rotate around a rotating center line and is connected with the host machine, the rotary driving device is used for driving the rotating seat to rotate, and the electrode assembly is detachably arranged on the rotating seat and can synchronously rotate with the rotating seat.

10. The flow electroporation device of claim 9, wherein: the rotating center line extends along the horizontal direction, a base line is arranged, the base line is a ray which is perpendicular to the rotating center line and extends vertically upwards, an end point of the base line is positioned on the rotating center line, the electrode assembly has a first limit position, a second limit position and a reference position in the rotating process of the electrode assembly around the rotating center line, and the rotating direction of the electrode assembly from the reference position to the second limit position is a positive direction;

when the electrode assembly is positioned at the reference position, the axial lead of the liquid outlet coincides with the base line;

when the electrode assembly is in the first limit position, the included angle between the axis line of the liquid outlet and the base line is-10 degrees;

when the electrode assembly is in the second limit position, an included angle between the axis line of the liquid outlet and the base line is 190 degrees.

11. The flow electroporation device of claim 10, wherein: the rotating seat and the host are provided with a first limiting structure therebetween, and the first limiting structure is used for limiting the electrode assembly to rotate between the first limiting position and the second limiting position.

12. The flow electroporation device of claim 11, wherein: the host machine is provided with a supporting plate extending along the up-down direction, and the supporting plate is provided with a limiting hole; the rotating seat is provided with a connecting part, the rotating center line passes through the connecting part, the connecting part is provided with a convex part protruding outwards along the radial direction of the rotating center line, and the connecting part is inserted into the limiting hole; the first limiting structure comprises the convex part and the limiting hole, and a first boss and a second boss which are arranged at intervals are arranged on the side wall of the limiting hole along the circumferential direction of the rotation center line;

when the electrode assembly is at the first limit position, the convex part is abutted against the first boss;

when the electrode assembly is in the second limit position, the convex part abuts against the second boss.

13. The flow electroporation device of claim 9, wherein: the host computer has first lateral wall, rotate the seat can around rotate the central line relative rotation ground with first lateral wall is connected, rotate the central line perpendicular to first lateral wall, rotate the seat including rotating the platform, follow rotate the extending direction of central line, rotate the platform with be equipped with the clearance between the first lateral wall, the inlet with the liquid outlet all is located rotate the platform keep away from one side of first lateral wall.

14. The flow electroporation device of claim 13, wherein: the rotating seat further comprises a connecting seat, the connecting seat penetrates through the first side wall along the extending direction of the rotating center line, the rotating driving device is arranged on one side of the first side wall, the rotating table is arranged on the other side of the first side wall, a second limiting structure is arranged between the host machine and the connecting seat and used for limiting the rotating table to move towards the first side wall.

15. The flow electroporation device of claim 14, wherein: the connecting seat is provided with an annular convex surface, and the extending direction of the convex surface is perpendicular to the rotation center line; the second limit structure comprises a plurality of limit pieces and convex surfaces, the limit pieces are arranged along the circumferential direction of the connecting seat at intervals, the limit pieces and the rotating table are respectively arranged on the two different sides of the first side wall, and each limit piece is propped against the convex surface.

16. The flow electroporation device of claim 15, wherein: the connecting seat is provided with a first part and a second part which are connected along the extending direction of the rotation center line, the outer diameter of the first part is larger than that of the second part, one side of the first part, which faces the second part, is the convex surface, each limiting part can be arranged on the first side wall in a relatively rotating way around the axis of the limiting part, the outer peripheral surface of each limiting part is propped against the outer peripheral surface of the second part, and the limiting parts are in rolling contact with the second part.

17. The flow electroporation device of claim 9, wherein: the flow electroporation device further comprises a limiting piece which can be connected with the host machine in a relative rotation mode around the axis of the flow electroporation device, and a plurality of limiting pieces are arranged at intervals; the rotating seat further comprises a connecting seat, and the limiting piece is in rolling contact with the outer peripheral surface of the connecting seat.

18. The flow electroporation device of claim 17, wherein: the host machine is provided with a first side wall, the rotating seat can rotate around the rotating center line relatively to be connected with the first side wall, the limiting piece is connected with the first side wall, and the connecting seat penetrates through the first side wall along the extending direction of the rotating center line.