CN217651240U

CN217651240U - Flow type electroporation device

Info

Publication number: CN217651240U
Application number: CN202221833842.1U
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: 2022-10-25
Anticipated expiration: 2032-07-14

Abstract

The utility model discloses a STREAMING electroporation device, include the host computer and locate electrode subassembly on it, advance kind container and collect the container. The electrode assembly comprises an electrode for performing electroporation treatment on 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 rotating manner; the sample introduction 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 processed by the electrode and 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 carries out electroporation treatment on cell suspension; the electrode assembly rotates relative to the host machine to be converted into a second working state; in the second working 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 utility model discloses can reduce the bubble when cell processing and produce, improve cell treatment effeciency, improve the cell rate of recovery after handling.

Description

Flow type electroporation device

Technical Field

The utility model relates to the field of biotechnology, especially, relate to a STREAMING electroporation device.

Background

Electroporation (Electroporation) is a technique that uses an electric field to act on a cell membrane to make the cell membrane generate micropores through which exogenous molecules such as DNA, RNA, and protein can pass, and these micropores are generated instantaneously under the action of the electric field and can recover with the disappearance of the electric field, thereby avoiding permanent damage to the cell. The electroporation technology can introduce various exogenous molecules such as DNA, RNA, protein, saccharides, dyes, virus particles and the like into prokaryotic or eukaryotic cells, is called as the Electrotransfection technology (Electrotransfection), has wide application in various high-end biomedical technical fields such as antibody protein production, in Vitro Diagnosis (IVD) reagent raw material production, gene editing and gene modification of immune cells and the like, and is an important technology for delivering substances in cells.

There are various methods for performing electroporation treatment on cells, wherein the flow electroporation technique can perform continuous electroporation treatment on flowing cell-containing suspension (hereinafter referred to as cell suspension), is relatively efficient, and is particularly suitable for batch treatment of large-volume cell suspension. A conventional flow electroporation apparatus mainly includes a pair of planar electrodes for generating an electric field, and a chamber located between the two planar electrodes, and a cell suspension can be electrically shocked while flowing through the chamber to generate electroporation, so that exogenous molecules in the cell suspension can enter target cells. In the prior art, most of flow electroporation devices have the problems of low cell treatment efficiency, large residual amount of cell suspension and the like. On one hand, the cell suspension can generate water electrolysis phenomenon when receiving electric shock, so that a large amount of bubbles are easily generated, the problems of discontinuous or uneven liquid flow, unstable flow rate and the like are caused, the uniformity of an electric field is reduced, and the controllability of the number of times of receiving the electric shock by the cells is influenced, so that the cell transfection efficiency is reduced; on the other hand, bubbles generated in the electroporation process can form foams, and the mixture of the foams and cells easily causes cell suspension to be retained in the chamber and the pipeline and is difficult to drain, so that the final cell recovery rate of the flow electroporation device is reduced.

SUMMERY OF THE UTILITY MODEL

The utility model aims at the problem that prior art exists, provide a flow type electroporation device that cell treatment efficiency is high.

In order to achieve the above purpose, the utility model adopts the technical scheme that:

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

the electrode assembly comprises an electrode for performing electroporation treatment on 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 rotating manner;

the sample introduction 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 carries out electroporation treatment on the cell suspension; during the process of converting the flow electroporation device from the first operating state to the second operating state, the electrode assembly rotates relative to the main machine; when the flow electroporation device is in the second working state, at least part of the cell suspension in the electrode assembly can flow out of the liquid outlet under the action of gravity.

In some embodiments, when the flow electroporation device is in the first operating state, the liquid inlet has a height that is lower than a height of the liquid outlet; when the flow electroporation device is in the second working state, the height of the liquid outlet is reduced. Under first operating condition, STREAMING electroporation device can normal operating, and the cell suspension is followed the flow direction of advancing upward play and is continuously flowed through electrode subassembly and carry out the electroporation and handle, can be full of the cavity between the electrode slice under the effect of cell suspension self gravity, keeps electric UNICOM between the electrode slice, has improved the continuity and the homogeneity of liquid stream, effectively reduces the bubble and produces, has improved the homogeneity of electric field again to the electroporation efficiency has been synthesized. When the second working state is in, the height of the liquid outlet is reduced, so that the cell suspension which is not completely 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 liquid outlet is at a lower height than the liquid inlet. The electrode assembly thus has a greater angle of inclination to facilitate more complete drainage of the cell suspension.

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

In some embodiments, the electrodes include a first electrode and a second electrode disposed at an interval, a processing channel is disposed between the first electrode and the second electrode, the processing channel extends in an up-down direction when the flow electroporation device is in the first operating state, and the cell suspension flows through the processing channel from bottom to top. Therefore, the cell suspension can not only enter and exit from the liquid inlet and the liquid outlet of the whole electrode assembly, but also keep the flowing direction of the entering and the exiting from the bottom to the top in the specific processing channel between the electrodes, and further ensure the uniformity of liquid flow and the uniformity of electric field in the cavity between the electrodes.

In some embodiments, the flow electroporation apparatus further comprises a liquid level sensor disposed between the sample introduction container and the electrode assembly, the liquid level sensor is fixedly disposed at 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 has a pipeline guide groove, which can be tightly attached to a liquid inlet pipeline, thereby facilitating the normal operation of the liquid level sensor. The pipeline guide groove extends along the vertical direction, and the liquid inlet pipeline penetrates 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 a signal to the control system, and the control system controls the working state of the electrode assembly. The liquid level sensor adopts a working mode that the liquid level sensor is vertically placed and cell suspension in the sample introduction pipeline enters from bottom to top, so that the liquid inlet pipeline in the pipeline guide groove can be full of liquid, the misjudgment of the detection result caused by the fact that the liquid in the pipeline is not full or bubbles is avoided, and the correct control of the control system on the electroporation process is influenced.

In some embodiments, 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. The peristaltic pump is used for providing power for the cell suspension to flow in the whole flow type electroporation device, the peristaltic pump is arranged between the collection container and the electrode assembly and is arranged at the rear end and below the electrode assembly, and the peristaltic pump is combined with the liquid outlet pipeline of the electrode assembly more smoothly. When the peristaltic pump is started, negative pressure is formed in the electrode assembly and a pipeline on the front side of the electrode assembly to suck and flow out cell suspension in the sample introduction container; when the whole pipeline is filled with liquid, the internal pressure of the pipeline is balanced, and the peristaltic pump stably transmits the cell suspension. Compare and install on the feed liquor pipeline and form malleation, promote the cell suspension flow in the pipeline of electrode subassembly and rear side in the peristaltic pump, the utility model discloses the velocity of flow of well cell suspension is more even stable, helps obtaining better electroporation treatment effect.

In some embodiments, the flow electroporation device further comprises a liquid level sensor and a peristaltic pump, the main machine 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 intersects with the extending direction of the second side wall, and the extending direction of the first side wall intersects with the extending direction of the third side wall; the electrode assembly is arranged on the first side wall in a relatively rotating mode, and the liquid level sensor and the peristaltic pump are arranged on the first side wall; the sample introduction container is arranged on the second side wall, and the collection 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 pipelines are more neatly arranged, the rotation of the electrode assembly is not easy to cause the crossed winding of the pipelines, and the smooth liquid flow is ensured; the sampling container and the collecting 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 fixed, the influence of the rotation of the electrode assembly is not easy to receive, the width of the host is favorably shortened, and the occupied space of the host is reduced.

In some embodiments, the flow electroporation apparatus further comprises a rotating base and a rotation driving device, the rotating base is connected with the host machine in a relatively rotating manner around a rotating 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, so that the stability and the reliability of the electrode assembly during rotation are improved, and the electrode assembly and the rotating seat move synchronously.

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, the 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 direction of the electrode assembly rotating from the reference position to the second limit position is a positive direction; when the electrode assembly is at the reference position, the axis of the liquid outlet is coincident with the base line; when the electrode assembly is at the first limit position, the included angle between the axial lead of the liquid outlet and the base line is-10 degrees; when the electrode assembly is at the second limit position, the included angle between the axial lead of the liquid outlet and the base line is 190 degrees.

In some embodiments, a first limiting structure is disposed between the rotating seat and the main frame, and the first limiting structure is configured to limit the rotation of the electrode assembly between the first limit position and the second limit position. The first limit structure limits the rotation angle of the electrode assembly, and the electrode assembly can only rotate between preset limit positions. Generally, in the first operating condition, the electrode assembly is in a reference position; under 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, the cell suspension remained in the pipeline near the liquid outlet of the electrode assembly can be well drained. The first limiting structure effectively avoids the problem that pipelines possibly lead to overlarge rotation angles are pulled or knotted, and the running reliability of the flow type electroporation device is improved.

In some embodiments, the host is provided with a support plate extending in the up-down direction, and the support plate is provided with a limit hole; the rotating seat is provided with a connecting part, the rotating center line penetrates through the connecting part, the connecting part is provided with a convex part which protrudes 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 the side wall of the limiting hole is provided with a first boss and a second boss which are arranged at intervals along the circumferential direction of the rotating central line; the projection abuts the first boss when the electrode assembly is in the first extreme position; the projection abuts the second boss when the electrode assembly is in the second extreme position. The first limiting structure specifically adopts a structure that the simple convex part is matched with the two groups of bosses, and has the advantages of simple design, no occupied space and strong reliability.

In some embodiments, the host computer has a first lateral wall, the rotation seat can wind the rotation center line relatively rotate ground with the first lateral wall is connected, the rotation center line perpendicular to the first lateral wall, the rotation seat includes the revolving stage, follows the extending direction of rotation center line, the revolving stage with be equipped with the clearance between the first lateral wall, the inlet with the liquid outlet all is located the revolving stage keep away from one side of first lateral wall. The rotating table provides a supporting plane for the rotation of the electrode assembly, and 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 does not protrude excessively and occupy space, and meanwhile, the friction between the rotating table and the first side wall can be reduced, and the smoothness 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 disposed on one side of the first side wall, the rotating platform is disposed on the other side of the first side wall, a second limiting structure is disposed between the main machine and the connecting base, and the second limiting structure is used for limiting the rotating platform to move towards the first side wall. The second limiting structure can be used for keeping the gap, and the rotating table is prevented from moving along the axial direction in the rotating process of the rotating seat.

In some embodiments, the connecting seat has a ring-shaped convex surface, and the extension direction of the convex surface is perpendicular to the rotation center line; the second limiting structure comprises a plurality of limiting parts and convex surfaces, the limiting parts are arranged along the circumferential direction of the connecting seat at intervals, the limiting parts and the rotating platform are respectively arranged on two different sides of the first side wall, and each limiting part is abutted against the convex surfaces. The second limiting structure mainly prevents the rotating platform from moving towards the inner side of the first side wall in a mode of pushing the connecting seat outwards from 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, one side of the first portion facing the second portion is the convex surface, each of the position-limiting members is disposed on the first sidewall in a manner of being capable of rotating relatively around its own axis, an outer circumferential surface of each of the position-limiting members abuts against an outer circumferential surface of the second portion, and the position-limiting members are in rolling contact with the second portion. The locating part can be spacing to the second portion of connecting seat in circumference to the locating part adopts rolling friction's mode and second portion contact, helps reducing spacing friction that causes, improves and rotates seat pivoted smoothness nature.

In some embodiments, the flow electroporation apparatus further comprises a limiting member connected to the main frame in a manner of being capable of rotating around its axis, and a plurality of the limiting members are disposed at intervals; the rotating seat further comprises a connecting seat, and the limiting part is in rolling contact with the outer peripheral surface of the connecting seat. The limiting parts can be used for providing support and rotation guide for the rotating seat.

In some embodiments, the main body has a first side wall, the rotating base is connected to the first side wall in a manner of relatively rotating around the rotation center line, the limiting member is connected to the first side wall, and the connecting base passes through the first side wall along the extending direction of the rotation center line. The rotating base and the limiting part are connected with the first side wall, and the supporting and guiding functions of the limiting part are more stable and reliable.

Because above-mentioned technical scheme's application, the utility model provides a stream type electroporation device realizes the switching between first operating condition and the second operating condition through the rotation of electrode subassembly for the host computer to can change the orientation of liquid outlet as required, after cell suspension was handled by the electroporation, its remaining part or whole cell suspension can only lean on self gravity discharge electrode subassembly, improve the cell rate of recovery.

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 below.

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

FIG. 2 is a schematic front view of the flow electroporation device of this embodiment in a second operating state;

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

fig. 4 is an assembled perspective view of the electrode assembly, the rotary base and the rotary motor in the present embodiment, in which the flow electroporation apparatus is in a first working state;

FIG. 5 is a perspective view of the electrode assembly, the rotary base and the rotary motor in this embodiment, in which the flow electroporation apparatus is in a second operating state;

FIG. 6 is a schematic perspective view of a portion of a flow electroporation apparatus according to an embodiment;

FIG. 7 is a partially exploded view of the flow electroporation apparatus according to this embodiment;

fig. 8 is a rear view schematically illustrating the rotary base and the motor bracket according to the present embodiment;

wherein: 100. a host; 110. a first side wall; 111. a control panel; 120. a second side wall; 121. a first bracket; 130. a third side wall; 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 access 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. mounting grooves; 620. a rotating table; 630. a connecting seat; 631. a first part; 632. a second section; 633. a limiting member; 640. a rotating seat main body; 641. a connecting portion; 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. a center line of rotation; 1002. a baseline; 1003. the axial lead of the liquid outlet.

Detailed Description

The following detailed description of the preferred embodiments of the invention, taken in conjunction with the accompanying drawings, enables the advantages and features of the invention to be more readily understood by those skilled in the art.

Referring to fig. 1-8, a flow electroporation apparatus for electroporating a cell suspension is shown. The flow electroporation device comprises a main machine 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, wherein the electrode assembly 200, the sample introduction container 310, the collection container 320, the liquid level sensor 400, the peristaltic pump 500, the rotary seat 600 and the rotary driving device are arranged on the main machine 100.

Referring to fig. 1 and 2, in the present embodiment, the main unit 100 is substantially rectangular, and with reference to the direction shown in the figure, the main unit 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, 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. Actually, the first sidewall 110, the second sidewall 120, and the third sidewall 130 respectively extend along a vertical direction, the first sidewall 110 is perpendicular to the second sidewall 120 and the third sidewall 130 respectively, the second sidewall 120 is parallel to the third sidewall 130, the upper portion of the first sidewall 110 is a control panel 111, and a rotation driving device and a control system are disposed inside the host 100.

In this embodiment, the second sidewall 120 is provided with a first bracket 121, and the first bracket 121 is used for suspending the sample container 310; the third sidewall 130 is provided with a second bracket 131, the second bracket 131 is used for suspending the collection container 320, the first bracket 121 is relatively located above the second bracket 131, the cell suspension can be transmitted from the sample injection container 310 to the collection container 320, and the sample injection container 310 and the collection container 320 are both arranged outside the corresponding sidewalls.

In this embodiment, the electrode assembly 200, the liquid level sensor 400, and the peristaltic pump 500 are disposed outside the first sidewall 110. The electrode assembly 200 includes a housing 230 having an inner cavity, an electrode (not shown) for performing electroporation treatment on the cell suspension is disposed in the inner cavity, the electrode assembly 200 further has a liquid inlet 210 and a liquid outlet 220 respectively disposed on the housing 230, and the liquid inlet 210 and the liquid outlet 220 are respectively disposed on two 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 is perpendicular to the first sidewall 110 and extends in a horizontal direction. Further, the electrodes comprise a first electrode and a second electrode which are arranged at intervals, a processing 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 the cell suspension can be subjected to electroporation processing when flowing through the processing channel.

In this embodiment, the sample container 310 is used for storing a cell suspension to be processed by an electrode, the lower portion of the sample container 310 has a first channel port 311, and the first channel port 311 is communicated with the liquid inlet 210 of the electrode assembly 200 through a liquid inlet pipeline 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 opening 321, and the second opening 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 container 310 and the electrode assembly 200, the liquid level sensor 400 has a pipeline guide slot 401, the pipeline guide slot 401 extends along a vertical direction, and the liquid inlet pipeline 312 passes through the pipeline guide slot 401 from bottom to top. The 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 line 312 and feed back a signal to the control system, which controls the operation state of the electrode assembly 200. Peristaltic pump 500 is disposed between collection container 320 and electrode assembly 200, with outlet line 322 passing through peristaltic pump 500 from top to bottom.

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

The flow electroporation device has a first working state and a second working state under the drive of the rotary drive device. Referring to fig. 1 and 4, when the flow electroporation apparatus is in the first operating state, the electrodes in the electrode assembly 200 can perform electroporation on the cell suspension, at this time, the height of the liquid inlet 210 is lower than the height of the liquid outlet 220, the axial line of the liquid inlet 210 and the axial line of the liquid outlet 220 both extend in the vertical direction, the two are located in the same vertical plane perpendicular to the first sidewall 110, at this time, the processing channel between the electrodes also extends in the up-down direction, and the cell suspension can flow through the processing channel from bottom to top and receive electric shock. So can ensure that whole processing channel is full of by cell suspension, and cell suspension velocity of flow is even, and the bubble that produces in the electroporation process simultaneously can discharge through the liquid outlet 220 at casing 230 top fast, can not be in the inside detention of processing channel and influence the electroporation effect.

Referring to fig. 2 and 5, when the flow electroporation apparatus is in the second operating 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, in this embodiment, the height of the liquid outlet 220 in the second operating state is lower than the height of the liquid inlet 210. Specifically, compared to the first operating state, the liquid outlet 220 rotates about 150 ° clockwise around the rotation center line 1001 and then is switched to the second operating state. Because the cross-sectional area of the processing channel is larger than that of the liquid outlet pipeline 322, a part of cell suspension remained in the processing channel can not be discharged due to the small surface tension of the cell suspension and the dead weight of the cell suspension after the electroporation processing, and the cell loss is caused. At this time, after the rotation seat 600 and the electrode assembly 200 are rotated by a certain angle by the rotation driving device, the liquid outlet 220 is lowered, so that the cell suspension retained in the processing channel is sufficiently discharged under the combined action of gravity and the suction force of the peristaltic pump 500, and is collected in the collection container 320.

It should be noted that, in the present embodiment, when the flow electroporation apparatus is in the first operating state and the second operating state, the peristaltic pump 500 is always operated, so that the cell suspension can be continuously and sufficiently sucked out from the electrode assembly 200 in cooperation with the gravity of the cell suspension itself. In other embodiments, the 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 the electrode assembly 200 by gravity alone.

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

Referring to fig. 6 and 7, in the embodiment, the rotation driving device specifically employs a rotation motor 710, a motor bracket 720 is fixedly disposed on the main body 100, the motor bracket 720 is specifically and fixedly disposed on the inner side of the first sidewall 110, the motor bracket 720 includes a supporting plate 721 extending along the vertical direction and a connecting plate 722 extending along the horizontal direction, the upper portion of the supporting plate 721 is fixedly disposed with the rear portion of the connecting plate 722, the front portion of the connecting plate 722 is fixedly disposed on the first sidewall 110, and a limiting hole 723 is disposed on the supporting plate 721. The rotating electric machine 710 includes a motor main body 711 and an output shaft 712, and the output shaft 712 is provided on the motor main body 711 so as to be relatively rotatable around a rotation center line 1001. The motor main body 711 is fixedly disposed at one side of the supporting plate 721, the rotating base 600 is relatively rotatably disposed at the other side of the supporting plate 721, specifically, the rotating base 600 is disposed at one side close to the first sidewall 110, the motor main body 711 is disposed at one side far away from the first sidewall 110, the output shaft 712 penetrates through the limiting hole 723 and is connected to the connecting portion 641 of the rotating base 600, and the output shaft 712 can drive the entire rotating base 600 to rotate.

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

Referring to fig. 3, in order to ensure that the cell suspension can only flow into the electrode assembly 200 in the predetermined direction and ensure the cell processing effect, a third limiting structure is disposed between the electrode assembly 200 and the rotating base 600 for limiting the electrode assembly 200 to be mounted on the rotating base 600 only in the predetermined direction.

Referring to fig. 4 and 5, in order to improve the reliability of the device, the connection holder 630 and the electrode assembly 200 in this embodiment do not rotate 360 ° around the rotation center line 1001, but have a first limit position and a second limit position of rotation. Specifically, first, the position of the electrode assembly 200 in the first operation state of the flow electroporation apparatus is defined as a reference position, a virtual base line 1002 exists, the base line 1002 is a ray extending vertically upward and perpendicular to the rotation center line 1001, and the 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 end position is defined as a positive direction (clockwise rotation is shown in the figure as a positive direction). When the electrode assembly 200 is at the reference position, the axis 1003 of the liquid outlet is coincident with the baseline 1002; when the electrode assembly 200 is in the first extreme position, the included angle between the axis 1003 of the liquid outlet and the base line 1002 is-10 degrees, namely, the liquid outlet 220 is slightly inclined to the upper left; when electrode assembly 200 is in the second extreme position, the angle between port axis 1003 and base 1002 is 190 °, i.e. port 220 is slightly tilted to the lower left.

Referring to fig. 7 and 8, a first limiting structure is disposed between the rotary base 600 and the main body 100, and the first limiting structure is used to limit the electrode assembly 200 to only rotate between the first limiting position and the second limiting position, that is, only to rotate from the first limiting position to the second limiting position in the forward direction, or to rotate from the second limiting position to the first limiting position in the reverse direction. It should be noted that the normal operation of the flow electroporation apparatus mainly involves the aforementioned first operation state and second operation state, the electrode assembly 200 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 position-limiting structure includes a protrusion 642 and a position-limiting hole 723. The tail of the rotating base main body 640 has the connecting portion 641, the rotation center line 1001 passes through the connecting portion 641, the protruding portion 642 is fixed on the connecting portion 641 and protrudes outward along the radial direction of the rotation center line 1001, and the connecting portion 641 and the protruding portion 642 are inserted into the limiting hole 723. Along the circumferential direction of the rotation center line 1001, the sidewall of the limiting hole 723 has a first boss 723a and a second boss 723b arranged at an interval, and the central angle included angle between the first boss 723a and the second boss 723b is about 200 °. When electrode assembly 200 is in the first limit position, boss 642 abuts against first boss 723 a; when the electrode assembly 200 is in the second limit 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 platform 620 is disposed at an interval with the first sidewall 110, and the liquid inlet 210 and the liquid outlet 220 are both exposed outside the mounting groove 610 and located on a side of the rotation platform 620 away from the first sidewall 110. A gap of about 0.5-0.8 mm can be kept between the rotating table 620 and the first side wall 110, and the direct contact friction between the rotating table and the first side wall and the influence on the rotation are avoided. In order to maintain the above gap, a second limiting structure is disposed between the main body 100 and the connecting seat 630, and the second limiting structure is used for limiting the movement of the rotating table 620 towards the first sidewall 110. Specifically, the connecting seat 630 has a first portion 631 and a second portion 632 connected along an extending direction of the rotation center line 1001, an outer diameter of the first portion 631 is larger than an outer diameter of the second portion 632, so that one side of the first portion 631 facing the second portion 632 has an annular convex surface, and an extending direction of the convex surface is perpendicular to the rotation center line 1001. The second limiting structure includes a plurality of limiting members 633, the limiting members 633 are disposed at intervals along the circumferential direction of the second portion 632, the limiting members 633 are specifically rollers, each limiting member 633 can be disposed on the first sidewall 110 around its axis line in a relatively rotating manner, the axis line of the limiting member 633 is parallel to the rotation center line 1001, and the limiting members 633 and the rotation driving device are located on the same side of the first sidewall 110, that is, located inside the first sidewall 110. One axial side surface of each of the stoppers 633 abuts against the annular convex surface, the outer circumferential surface of each of the stoppers 633 abuts against the outer circumferential surface of the second portion 632, and the stoppers 633 are in rolling contact with the second portion 632. In this embodiment, the limiting members 633 are six and are uniformly spaced along the circumferential direction of the second portion 632, so as to not only limit the axial movement of the rotating table 620, but also provide rotational guidance for the limiting members 633, and assist in supporting the rotating base 600, thereby reducing the load of the rotating electrical machine 710 to a certain extent.

In this embodiment, in order to avoid the poor rotation or noise caused by insufficient torque of the rotating electric machine 710, the device is designed to be light, and the load of the rotating electric machine 710 should not exceed 1kg. Specifically, the rotating base 600 is made of a light material, and has a weight of about 0.4kg, and since the limiting member 633 bears a part of the weight of the rotating base 600, the rotating electric machine 710 actually bears a lower load, thereby being capable of stably operating.

In conclusion, the flow electroporation device provided by the embodiment can significantly improve the uniformity and continuity of cell suspension flow in the electroporation process and improve the electroporation efficiency, can fully discharge the cell suspension after treatment, and improves the 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 the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and the protection scope of the present invention can not be limited thereby, and all equivalent changes or modifications made according to the spirit of the present invention should be covered by the protection scope of the present invention.

Claims

1. A flow electroporation device, comprising: the flow electroporation device comprises a host machine, an electrode assembly, a sample introduction container and a collection container, wherein the electrode assembly, the sample introduction container and the collection container are respectively arranged on the host machine,

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

3. A flow electroporation device as claimed in 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. A flow electroporation device as claimed in 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 both extend in the vertical direction.

5. A flow electroporation device as claimed in claim 1, wherein: the flow-type electroporation device is in the first working state, the processing channel extends along the up-down direction, and the cell suspension flows through the processing channel from bottom to top.

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

7. A flow electroporation device as claimed in claim 1, wherein: the flow type electroporation device further comprises a peristaltic pump, the peristaltic pump is fixedly arranged on one side of the host machine, and the liquid outlet pipeline penetrates through the peristaltic pump from top to bottom.

8. A flow electroporation device as claimed in claim 1, wherein: the flow electroporation device further comprises a liquid level sensor and a peristaltic pump, the host machine 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 in 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 is connected with the first side wall in a relatively rotatable mode, and the liquid level sensor and the peristaltic pump are arranged on the first side wall; the sample introduction container is arranged on the second side wall, and the collection container is arranged on the third side wall.

9. A flow electroporation device as claimed in claim 1, wherein: the flow type electroporation device further comprises a rotating seat and a rotating driving device, the rotating seat can be connected with the host machine in a relatively rotating mode around a rotating center line, the rotating driving device is used for driving the rotating seat to rotate, and the electrode assembly is detachably arranged on the rotating seat and can rotate synchronously with the rotating seat.

10. A flow electroporation device as claimed in claim 9, wherein: the rotation center line extends along the horizontal direction, a base line is arranged, the base line is a ray which is perpendicular to the rotation center line and extends vertically upwards, the end point of the base line is positioned 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 direction of the electrode assembly rotating from the reference position to the second limit position is a positive direction;

when the electrode assembly is at the reference position, the axis of the liquid outlet is coincident with the base line;

when the electrode assembly is positioned at the first limit position, the included angle between the axial lead of the liquid outlet and the base line is-10 degrees;

when the electrode assembly is at the second limit position, the included angle between the axial lead of the liquid outlet and the base line is 190 degrees.

11. A flow electroporation device as claimed in claim 10, wherein: and a first limiting structure is arranged between the rotating seat and the host machine and is used for limiting the electrode assembly to rotate between the first limiting position and the second limiting position.

12. A flow electroporation device as claimed in claim 11, wherein: the host is provided with a support plate extending along the vertical direction, and the support plate is provided with a limit hole; the rotating seat is provided with a connecting part, the rotating center line penetrates through the connecting part, the connecting part is provided with a convex part which protrudes outwards along the radial direction of the rotating center line, and the connecting part is inserted in the limiting hole; the first limiting structure comprises the convex part and the limiting hole, and the side wall of the limiting hole is provided with a first boss and a second boss which are arranged at intervals along the circumferential direction of the rotating central line;

the projection abuts the first boss when the electrode assembly is in the first extreme position;

the projection abuts the second boss when the electrode assembly is in the second extreme position.

13. A flow electroporation device as claimed in claim 9, wherein: the host computer has first lateral wall, rotate the seat and can wind center of rotation line relatively rotate ground with first lateral wall is connected, center of rotation line perpendicular to first lateral wall, rotate the seat including rotating the platform, follow center of rotation line's extending direction, rotating the platform with be equipped with the clearance between the first lateral wall, the inlet with the liquid outlet all is located rotating the platform keep away from one side of first lateral wall.

14. A flow electroporation device as claimed in 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 rotary driving device is arranged on one side of the first side wall, the rotating platform is arranged on the other side of the first side wall, a second limiting structure is arranged between the host and the connecting seat, and the second limiting structure is used for limiting the rotating platform to move towards the first side wall.

15. A flow electroporation device as claimed in claim 14, wherein: the connecting seat is provided with an annular convex surface, and the extending direction of the convex surface is vertical to the rotating central line; the second limiting structure comprises a plurality of limiting parts and convex surfaces, the limiting parts are arranged along the circumferential interval of the connecting seat, the limiting parts and the rotating platform are arranged on two different sides of the first side wall, and each limiting part is abutted against the convex surface.

16. A flow electroporation device as claimed in claim 15, wherein: the connecting seat has along first portion and the second portion that center of rotation line's extending direction meets, the external diameter of first portion is greater than the external diameter of second portion, first portion orientation one side of second portion is the convex surface, every the locating part all can set up around self axial lead relative rotation on the first lateral wall, every the outer peripheral face of locating part all supports and leans on the outer peripheral face of second portion, the locating part with second portion rolling contact.

17. A flow electroporation device as claimed in claim 9, wherein: the flow type electroporation device also comprises limiting pieces which can be relatively rotatably connected with the host machine around the axis of the flow type electroporation device, and the limiting pieces are arranged at intervals; the rotating seat further comprises a connecting seat, and the limiting part is in rolling contact with the outer peripheral surface of the connecting seat.

18. A flow electroporation device as claimed in claim 17, wherein: the host computer has first lateral wall, rotate the seat can wind rotation center line relatively rotate ground with first lateral wall is connected, the locating part with first lateral wall links to each other, the connecting seat is followed rotation center line's extending direction passes first lateral wall.