CN209243073U - A kind of intermittent streaming electrotransfection device - Google Patents
A kind of intermittent streaming electrotransfection device Download PDFInfo
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- CN209243073U CN209243073U CN201821454480.9U CN201821454480U CN209243073U CN 209243073 U CN209243073 U CN 209243073U CN 201821454480 U CN201821454480 U CN 201821454480U CN 209243073 U CN209243073 U CN 209243073U
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Abstract
The utility model discloses a kind of intermittent streaming electrotransfection devices, including tube body, first electrode, second electrode, first piston, it is characterized in that, the first electrode, second electrode is placed in the inner wall of the tube body in parallel, one end of the tube body seals, the first piston is located at the opening of the tube body, the inside of the first piston and tube body has the cavity for accommodating liquid sample, at least one liquid entrance is arranged in the tube body and at least one liquid passes in and out valve, the liquid disengaging valve is located at the tube body liquid entrance, control the opening and closing of liquid entrance, the first piston can be relatively moved along the tube body.Streaming electrotransfection device disclosed by the utility model can be improved the stability of electrotransfection device, improve electrotransfection efficiency.
Description
Technical field
The utility model relates to electrotransfection fields, are related to a kind of intermittent streaming electrotransfection device, more specifically, this reality
It is related to a kind of intermittent streaming electrotransfection device for inhibiting electrotransfection to generate bubble by way of the mobile pressurization of piston with novel.
Background technique
Cell membrane is the thin film for being enclosed in pericellular, be cell with the external world carry out selective substances exchange it is penetrating
Property barrier.Cell membrane makes cell become an independent living unit, and possesses a metastable interior environment.Ambient enviroment
In some substances can be by cell membrane, other substances are then not all right.Cell can be absorbed by cell membrane from ambient enviroment
Nutriment is discharged metabolite, the transhipment of substance is made to reach equilibrium state.So the basic function of cell membrane is exactly to maintain cell
Interior microenvironment relatively stable simultaneously selectively carries out mass exchange with external environment.
The study found that if applying the electro photoluminescence of some strength to cell and continuing for some time, so that it may inducing cell
Some micropores are generated on film, enhance the permeability of cell, and so-called cell electroporation (Electroporation) just refers to cell
Under the action of adding impulse electric field outside, the biophysics process of instantaneous micropore is formed on cell membrane lipid bilayer.Electrotransfection
It (Electrotransfection) is to be imported the large biological molecule of external source, such as DNA, RNA or protein using electroporation technology
The technology of cell.When electroporation occurs for cell membrane, permeability and membrane conductance can instantaneously increase, and make hydrophilic molecules, DNA, egg
White matter, virion, drug granule etc. cannot be able to enter cell by micropore by the molecule of cell membrane under normal circumstances.?
After removing electro photoluminescence in short time, micropore disappears on cell membrane, and cell membrane becomes selective permeation barrier again.
Compared with traditional chemical transfection and virus transfection, due to electrotransfection with no chemical contamination, cell will not be made
At permanent damage, efficiency is higher the advantages that, have in fields such as biophysics, molecular biology, clinical medicine wide
Application prospect.
Although the mechanism of electrotransfection effect is not entirely clear that, cell electrotransfection herein is well known, including thin
The movement of after birth lipid bilayer causes to form temporary micropore on film, and exogenous molecule is allowed to enter cell by micropore.
In the prior art, three classes method mainly completes the process of cell electrotransfection:
Cell is placed in a pair of between several millimeters to several centimeters parallel pole.Make the electricity of cell between the electrodes
By electro photoluminescence in, to realize the purpose of electrotransfection.For example, United States Patent (USP) US5389069.
It is penetrated in tissue or cell liquid using miniature needle electrode and is shocked by electricity to cell, achieve the purpose that electrotransfection.Example
Such as, United States Patent (USP) US5389069.
One chamber is placed between a pair of of parallel pole, while so that the aaerosol solution of cell flows in the chamber
It is shocked by electricity.For example, United States Patent (USP) US6773669.
Chinese patent CN201010242144 discloses a kind of streaming electrotransfection apparatus and system, which includes: streaming
Electrotransfection device, including: substrate, and the electrode being produced on substrate, the electrode are parallel and place in pairs
, each pair of electrode includes the anode and cathode being oppositely arranged；It is placed in the channel of the limitation fluid flowing on electrode；The channel
Starting point has multiple inlet branch channels, and pools a main channel, and end has multiple outlet branch channels, described
Setting has the top cover of multiple fluid inlets and outlet above channel；Syringe pump is connected to the streaming electrotransfection by pipeline and fills
Set the entrance of middle top cover and the flow velocity of control of export fluid；Voltage source is set by electrical connector connection electrode and generates pulse electricity
Pressure.Streaming electrotransfection system realizes various suspension in the fluid passage using fluid channel and the syringe pump being connected
Continuous flowing, so that cell be enable to be continued to carry out by the process of electrotransfection, realization quickly handles a large amount of samples.
Chinese patent CN201610806987 discloses the disposable product of the electrotransfection for cell comprising: in institute
State the fluid compartment of the inside of disposable product；First fluid port is used to provide cell suspension to the fluid compartment；With
And second fluid port, be used for will include to by the fluid delivery of electrotransfection at least one of cell compound extremely
The fluid compartment；First electrode and second electrode in the fluid compartment are set；At least one outlet port, delivering
The fluid from the fluid compartment, wherein the first fluid port and the second fluid port and hybrid channel are flowed
Body connection, the hybrid channel and the fluid compartment are in fluid communication.
But streaming electrotransfection device disclosed above can generate a large amount of bubbles during practical electrotransfection, generation
Bubble, which is attached on electrode plate, influences field uniformity, unstable so as to cause electrotransfection effect.Inventor passes through retrieval discovery,
The technical issues of electrotransfection process generates bubble is solved not yet in streaming electrotransfection field.
Summary of the invention
The utility model is provided a kind of by movably living for above-mentioned existing technology
The mode of pressurization is filled in, the intermittent streaming electrotransfection device of generation bubble during electrotransfection is inhibited.The utility model it is another
Purpose is to improve the stability of electrotransfection device, improves the efficiency of electrotransfection.
In order to achieve the above object, the technical solution of intermittent streaming electrotransfection device provided by the utility model is summarized
It is as follows: a kind of intermittent streaming electrotransfection device, including tube body, first electrode, second electrode, first piston, which is characterized in that
The first electrode, second electrode are placed in the inner wall of the tube body, one end sealing of the tube body, the first piston position in parallel
In the other end opening of the tube body, the inside of the first piston and tube body has the cavity for accommodating liquid sample, described
At least one liquid entrance is arranged in tube body and at least one liquid passes in and out valve, and the liquid disengaging valve is located at the tube body
At liquid entrance, the opening and closing of liquid entrance are controlled, the first piston can be relatively moved along the tube body.
Another technical solution of offer described in the utility model is summarized as follows: a kind of intermittent streaming electrotransfection device, packet
Include tube body, first electrode, second electrode, first piston, second piston, which is characterized in that the first electrode, second electrode are flat
Row is placed in the inner wall of the tube body, and the first piston and second piston are located at the opening at the tube body both ends, described
The inside of first piston, second piston and tube body has the cavity for accommodating liquid sample, at least one liquid is arranged in the tube body
Inlet and outlet and at least one liquid pass in and out valve, and liquid disengaging valve is located at the tube body liquid entrance, control into
The opening and closing of outlet, the first piston and/or second piston can be relatively moved along the tube body.
It is preferred that snorkel is arranged in the tube body, gas valve, the further preferred snorkel and tube body is arranged in snorkel
Gas valve is arranged in junction, and the snorkel is between the first piston and second piston or the snorkel position
Between the first piston and the sealed end of tube body, air filter is arranged in the end of the snorkel；
It is preferred that at least two liquid entrances are arranged in the tube body, the liquid entrance either liquid-inlet,
It can be liquid outlet, corresponding at least two liquid that are arranged pass in and out valve；Liquid disengaging valve either liquid into
Sample check valve is also possible to liquid discharge sample check valve；At least one liquid-inlet and one is arranged in the further preferred tube body
Liquid outlet, at least one Liquid sample introduction check valve and a liquid discharge sample check valve；
It is preferred that the cross section of the first piston and second piston and the cross section of the tubular body are identical or close,
Further preferably the shape of the cross section of the tubular body is rectangular, and round or other shapes also can be set into；
It is preferred that sealing device is arranged in the first piston and second piston and tube body contact position, the sealing device is preferred
Seal washer, sealing joint strip or other devices with sealing performance etc.；
It is preferred that when the liquid passes in and out valve and gas valve and closes, the first piston and tube body or first piston,
The inside of second piston and tube body forms closed cavity；
It is preferred that the first piston and/or second piston are opposite when the liquid passes in and out valve and gas valve is closed
After tube body is mobile, the pressure value of the closed inside cavity is 1.0-2.5Mpa；The pressure of the further preferred inside cavity
Force value is 1.1-2.0Mpa；The pressure value of the further preferred inside cavity is 1.2-1.5Mpa；
It is preferred that described device further includes that conduit is several, it is divided into inlet tube and outlet tube, conduit and tube body sealed set；
It is preferred that described device further includes pump, for letting the liquid out or flow into cavity, or for being discharged or being filled with cavity
Interior gas, the type of pump include but is not limited to peristaltic pump, air pump, liquid pump, syringe pump etc.；
It is preferred that described device further includes that the first piston and/or second piston is enable to relatively move along the tube body
Pressurizing device, the type of the pressurizing device includes but is not limited to air bag, hydraulic stem, atmospheric pressure pole, screw press, electromagnetism
Power pressurizing device etc.；
It is preferred that the first piston and/or second piston setting operating position and off-position, the piston is from non-work
It is moved to operating position as position, pressure can be provided for inside cavity；The further preferred liquid entrance is located at described
Between first piston and second piston；The further preferred liquid entrance is located at the sealed end of the first piston and tube body
Between.The further preferred liquid entrance is located at the first piston and/or second piston is arranged operating position and non-
Between operating position.
The preparation method of device described in the utility model: tube body, selection cured plastics work after capable of heating are prepared first
For the material of tube body, tube body is prepared using the mold for preparing tube body, Liquid sample introduction mouth and liquid discharge sample are respectively set on tube body
The electrode of two panels same shape, size is arranged in parallel in the opposite sides of inboard wall of tube body for mouth.Distinguish at the both ends open of tube body
One piston parallel relative to tube body end face is set, in piston and tube body contact position using sealing joint strip sealing, piston not
The side of contact cavity is connect with pressurizing device.Or block one end of tube body, it is arranged one in the other end opening of tube body
A piston parallel relative to tube body end face is sealed using sealing joint strip in piston and tube body contact position, does not contact chamber in piston
The side of body is connect with pressurizing device.
Technical solutions of the utility model bring the utility model has the advantages that
The intermittent streaming electrotransfection device of the utility model is capable of handling a large amount of fluid samples.
The intermittent streaming electrotransfection device of the utility model, to liquid pressing, inhibits bubble to produce during electrotransfection
Raw, electrotransfection efficiency fluctuation caused by reducing because of bubble attachment improves the stability of electrotransfection device.
The intermittent streaming electrotransfection apparatus structure of the utility model is novel in design, has initiative.
Detailed description of the invention
The intermittent streaming electrotransfection device exemplary diagram of Fig. 1 the utility model；The intermittent streaming electrotransfection of Fig. 2 the utility model
Device exemplary diagram；
Attached drawing annotation: 1- first electrode, 2- second electrode, 3- tube body, 4- inlet tube, 5- outlet tube, 6- pressurizing device, 7-
First piston, 8- second piston, 9- tube body sealing end, 10- sample introduction check valve, 11- go out sample check valve, 12- snorkel, 13- ventilation
Valve.
Specific embodiment
Embodiment one
Overall construction design (one)
Intermittent streaming electrotransfection device provided by the utility model is as shown in Figure 1, one end nozzle of tube body (3) is arranged
(9) are blocked at tube body, the first electrode (1) and second electrode (2) are arranged in parallel in the inner wall of tube body (3) respectively, and described the
One piston (7) is set at the other end nozzle of the tube body (3), the first piston (7) and pressurizing device (6) fixed company
It connects, first piston (7) can be moved to operating position, the preferred liquid of pressurizing device from off-position by pressurizing device (6)
Compression bar, the liquid for carrying inside cavity bear certain pressure.Snorkel (12) are arranged in the tube body (3), the snorkel
(12) gas valve (13) are set with tube body (3) junction.Apply electric pulse in first electrode (1) and second electrode (2), electricity
Pulse makes to form electric field in cavity, so that cell membrane forms micropore, so that the target substance in liquid sample be made to be able to enter carefully
In born of the same parents.After the completion of electrotransfection, first piston (7) can be moved to off-position from operating position by pressurizing device (6).It repeats
Aforesaid operations carry out intermittent streaming electrotransfection.
Embodiment two
Overall construction design (two)
Intermittent streaming electrotransfection device provided by the utility model is as shown in Fig. 2, the both ends nozzle of the tube body (3) is set
First piston (7) and second piston (8) are set, the first electrode (1) and second electrode (2) are arranged in parallel in the inner wall of tube body,
The first piston (7) and second piston (8) setting operating position and off-position, the first piston and second piston point
Be not fixedly connected with pressurizing device (6), pressurizing device (6) preferred hydraulic stem, pressurizing device (6) can by first piston (7) and
Second piston (8) is moved to operating position from off-position respectively, and the liquid of inside cavity is made to bear certain pressure.?
Applying electric pulse on one electrode (1) and second electrode (2), electric pulse makes to form electric field in cavity, so that cell membrane forms micropore,
To make the target substance in liquid sample be able to enter in cell.After the completion of electrotransfection, pressurizing device (6) can be living by first
Plug (7) and second piston (8) are moved to off-position from operating position respectively.It repeats aforesaid operations and carries out intermittent streaming electricity
Transfection.
Embodiment three
Liquid batch formula feed liquor mode and pressuring method
When feed liquor, sample introduction check valve (10) is opened, and sample check valve (11) is closed out, is opened gas valve (13), is used air pump
By snorkel (12) extraction and tube body (3) isometric gas, target liq sample enters inside cavity from inlet tube (4)；
Or without using air pump by snorkel (12) extraction and tube body (3) isometric gas, using peristaltic pump by target liq sample
This is pressed into inside cavity from inlet tube (4)；After cavity is full of, sample introduction check valve (10) and gas valve (13) are closed.By adding
Pressure device (6) applies specific pressure to first piston (7) and/or second piston (8), it is made to be moved to work from off-position
Make position, complete the pressurization to liquid, the electrotransfection of liquid is carried out in pressurized state.After the completion of electrotransfection, it is unidirectional to open out sample
Valve (11) and gas valve (13), are filled with the gas isometric with cavity, target liquid by snorkel (12) Xiang Guanti with air pump
Body sample is discharged from outlet tube (5)；Or the gas isometric with tube body is filled with to cavity by snorkel (12) without using air pump
Target liq sample is discharged from outlet tube (5) by gravity for body；After target liq sample is completely exhausted out, sample check valve out
(11) it closes, sample introduction check valve (10) is opened, and repeats the movement of the above feed liquor.
Example IV
Biological test experiment
The CHO-S cell (Chinese hamster ovary cell) for being in logarithmic growth phase is collected, 1000 revs/min of revolving speed are centrifuged 5 points
Clock abandons supernatant, and cell is resuspended with electrotransfection buffer, so that the density of cell is 1 × 107A/milliliter, addition need electrotransfection
It is transferred to the plasmid pCDNA3.1-GFP of cell, makes 20 mcg/ml of concentration of plasmid, it is soft to be uniformly mixed.
The electrotransfection device being prepared using the utility model embodiment one is tested, and the wriggling of complete set is passed through
Pump plus conduit device is blended with cell, the suspension of plasmid is transported in the cavity of electrotransfection device, utilizes hydraulic stem control the
One piston is mobile in operating position and off-position, and cooperation electrotransfection instrument uses, and carries out electro photoluminescence using following condition: electricity
180 volts, 6 milliseconds of pulse width, pulse number 2 times, the pulse spacing 1 second of pressure.
Comparative test is not apply pressure to liquid in cavity, and first piston is maintained at off-position, is filled out in liquid
After cavity, electrotransfection is directly carried out；
The test of technical solutions of the utility model is to apply pressure to liquid in cavity, after body is had one's bosom filled in liquid filling, is led to
It crosses hydraulic stem and first piston is moved to operating position from off-position, pressurize to liquid, carry out electrotransfection.
After electrotransfection, the cell suspension after transfection is placed in centrifuge tube, 1000 revs/min, is centrifuged 5 minutes.It abandons
Supernatant is added CD-OptiCHO culture medium and cell is resuspended, is inoculated in triangular pyramidal culture in glassware, culture density 2 × 106A/
Then triangular pyramidal bottle is placed on shaking table and cultivates by ml, 125 revs/min of shaking speed, condition of culture: temperature is 37 DEG C, dioxy
Change concentration of carbon 5%.After 24 hours fluorescence microscopy under the microscope, and with flow cytomery electrotransfection efficiency and cell survival
Rate.
By the observation to electrotransfection process, comparative test can generate a small amount of gas during electrotransfection near electrode
Bubble, influences electrotransfection effect；And the test of technical solutions of the utility model is during electrotransfection and bubble-free generates.
By the way that electrotransfection Comparative result, the electrotransfection efficiency of comparative test is 60%-63%, and cell survival rate is
72%-79%；The electrotransfection efficiency of technical solutions of the utility model test is 86%-89%, cell survival rate 80%-
82%.The experimental results showed that technical solutions of the utility model test provide electrotransfection device have higher electrotransfection efficiency and
Cell survival rate.
Although the utility model has been described in detail, the modification in the spirit and scope of the utility model is for ability
Field technique personnel will be apparent.It should be understood that sheet described above and/or in the appended claims is practical
The part of novel aspect and various embodiments and various features can be combined or completely or partially be interchangeable.Such as this
Field technical staff will be realized that, in the description of aforementioned various embodiments, refers to those of another embodiment and implements
Scheme can suitably with other combination of embodiment.In addition, it will be recognized by those of ordinary skill in the art that foregoing description is only to make
To illustrate, and it is not intended to limit the utility model.
Claims (18)
1. a kind of intermittent streaming electrotransfection device, including tube body, first electrode, second electrode, first piston, feature exist
In the first electrode, second electrode are placed in the inner wall of the tube body in parallel, and one end sealing of the tube body, described first is living
Plug is located at the other end opening of the tube body, and the inside of the first piston and tube body has the cavity for accommodating liquid sample,
At least one liquid entrance is arranged in the tube body and at least one liquid passes in and out valve, and the liquid disengaging valve is located at described
At tube body liquid entrance, the opening and closing of liquid entrance are controlled, the first piston can be along the opposite shifting of the tube body
It is dynamic.
2. intermittent streaming electrotransfection device according to claim 1, which is characterized in that the tube body setting at least two
Liquid entrance.
3. intermittent streaming electrotransfection device according to claim 1, which is characterized in that the first piston and/or
Operating position and off-position is arranged in two pistons.
4. intermittent streaming electrotransfection device according to claim 1, which is characterized in that snorkel is arranged in the tube body,
Gas valve is arranged in snorkel.
5. intermittent streaming electrotransfection device according to claim 4, which is characterized in that the snorkel is connect with tube body
Place's setting gas valve.
6. intermittent streaming electrotransfection device according to claim 1, which is characterized in that the first piston and second is lived
Sealing device is arranged with tube body contact position in plug.
7. intermittent streaming electrotransfection device according to claim 1, which is characterized in that when the liquid disengaging valve and
When gas valve is closed, the inside of the first piston and tube body or first piston, second piston and tube body forms closed chamber
Body.
8. intermittent streaming electrotransfection device according to claim 7, which is characterized in that when the liquid disengaging valve and
When gas valve is closed, the first piston and/or second piston are after tube body relative movement, in the closed cavity
The pressure value in portion is 1.0-2.50Mpa.
9. intermittent streaming electrotransfection device according to claim 1, which is characterized in that described device further include make it is described
The pressurizing device that first piston and/or second piston can be relatively moved along the tube body.
10. a kind of intermittent streaming electrotransfection device, including tube body, first electrode, second electrode, first piston, second piston,
It is characterized in that, the first electrode, second electrode are placed in the inner wall of the tube body, the first piston and second piston in parallel
It is located at the opening at the tube body both ends, the inside of the first piston, second piston and tube body, which has, accommodates liquid-like
This cavity, at least one liquid entrance is arranged in the tube body and at least one liquid passes in and out valve, and the liquid passes in and out valve
Door is located at the tube body liquid entrance, controls the opening and closing of inlet and outlet, the first piston and/or second piston energy
It is enough to be relatively moved along the tube body.
11. intermittent streaming electrotransfection device according to claim 10, which is characterized in that the tube body setting at least two
A liquid entrance.
12. intermittent streaming electrotransfection device according to claim 10, which is characterized in that the first piston and/or
Operating position and off-position is arranged in second piston.
13. intermittent streaming electrotransfection device according to claim 10, which is characterized in that the tube body setting ventilation
Gas valve is arranged in pipe, snorkel.
14. intermittent streaming electrotransfection device according to claim 13, which is characterized in that the snorkel and tube body connect
Connect place's setting gas valve.
15. intermittent streaming electrotransfection device according to claim 10, which is characterized in that the first piston and second
Sealing device is arranged in piston and tube body contact position.
16. intermittent streaming electrotransfection device according to claim 10, which is characterized in that when the liquid passes in and out valve
When closing with gas valve, the inside of the first piston and tube body or first piston, second piston and tube body forms closed
Cavity.
17. intermittent streaming electrotransfection device according to claim 16, which is characterized in that when the liquid passes in and out valve
When closing with gas valve, the first piston and/or second piston are after tube body relative movement, the closed cavity
Internal pressure value is 1.0-2.50Mpa.
18. intermittent streaming electrotransfection device according to claim 10, which is characterized in that described device further includes making institute
It states first piston and/or second piston can be along the pressurizing device of tube body relative movement.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201821454480.9U CN209243073U (en) | 2018-09-11 | 2018-09-11 | A kind of intermittent streaming electrotransfection device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201821454480.9U CN209243073U (en) | 2018-09-11 | 2018-09-11 | A kind of intermittent streaming electrotransfection device |
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| CN201821454480.9U Active CN209243073U (en) | 2018-09-11 | 2018-09-11 | A kind of intermittent streaming electrotransfection device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USD965170S1 (en) | 2020-10-23 | 2022-09-27 | Life Technologies Corporation | Electroporation device |
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2018
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USD965170S1 (en) | 2020-10-23 | 2022-09-27 | Life Technologies Corporation | Electroporation device |
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Address after: No.2 501, No.200, Shengpu Xingpu Road, Suzhou Industrial Park, Jiangsu Province Patentee after: SUZHOU ETTA BIOTECH CO., LTD. Address before: 215123 Ruoshui Road 388 E1504, Suzhou Industrial Park, Jiangsu Province Patentee before: SUZHOU ETTA BIOTECH CO., LTD. |
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