CN217173753U - Ring type transfection device utilizing six-hole plate - Google Patents

Abstract

The utility model discloses an utilize six orifice plates ring-type transfection device, including six orifice plate bodies, plate lid and electrode pairs, six orifice plate body intervals are equipped with six undercut electrotransfection cell and hold the chamber, the electrode pair sets up in pairs in each electrotransfection cell holds the intracavity and is connected with the power through the wire, and each electrode pair includes positive electrode and negative electrode, the positive electrode is the annular, the negative electrode is circular sheet structure, the negative electrode is located in the annular positive electrode, the plate lid detachable cover is in on six orifice plate bodies; this application once can carry out the electrotransfection to six or six kinds of cells simultaneously, and the electrotransfection is efficient, has solved a large amount of sample repetitive operation's problem, and the survival rate of cell is high moreover, and transfection effect is good, has fine using value.

Description

Ring type transfection device utilizing six-hole plate

Technical Field

The utility model relates to a cell transfection technical field, concretely relates to utilize six orifice plate ring type transfection devices.

Background

The cell membrane is a layer of thin film which surrounds the outer layer of the cell and is a barrier for the exchange of substances and information between the cell and the outside. Because of the cell membrane, the cell becomes an independent living individual. Cells can selectively take up nutrients through the cell membrane, excluding metabolites. The main components of the cell membrane are lipid, protein, saccharide, etc., and the total thickness is about 7-10nm, wherein the lipid is mainly phospholipid.

The research shows that if certain intensity of electric stimulation is applied to the cells for a period of time, micropores can be induced on cell membranes, so that the permeability of the cells is enhanced; the cell Electroporation (Electroporation) refers to the biophysical process of the cell forming transient micropores on the cell membrane lipid bilayer under the action of an applied pulse electric field. Electrotransfection (Electrotransfection) is a technique for introducing foreign biological macromolecules, such as DNA, RNA or proteins, into cells using electroporation. When the cell membrane is electroporated, the permeability and membrane conductance are increased instantaneously, so that molecules, such as hydrophilic molecules, DNA, proteins, virus particles, drug particles, and the like, which cannot pass through the cell membrane under normal conditions, can enter the cell through the micropores. After the electrical stimulation is removed in a short time, the micropores in the cell membrane disappear, and the cell membrane becomes a selective permeability barrier again. Compared with the traditional method (virus and chemistry), the electrotransfection technology has the advantages of no pollution, easier control, no permanent damage to cells, high efficiency and the like, and is widely applied to the fields of molecular biology, clinical medicine and the like.

In the prior art, there are mainly the following cell transfection methods:

1. the cells are placed between a pair of parallel electrodes spaced from each other by a distance of several millimeters to several centimeters, so that the cells are stimulated by electric pulses between the electric fields generated by the two electrodes, thereby realizing the transfection process. For example US patent 5389069.

2. The micro needle electrode is pricked into tissue or cell fluid to stimulate cell with electric pulse so as to reach the aim of electrotransfection. For example US patent 5389069.

3. A pair of electrodes is arranged in a cavity, so that the cell suspension is electrically stimulated while flowing in the cavity, and the purpose of electrotransfection is achieved. For example US 6773669.

At present, electrotransfection is mostly carried out in a single test tube, and the maximum volume of electrotransfection is about 1ml, so that when a large amount of samples are processed, the problem of high labor intensity is faced.

SUMMERY OF THE UTILITY MODEL

To the defects in the prior art, the present invention aims to provide a ring-type transfection device using six-hole plates, which can be used for high-efficiency transfection of various cells.

The utility model adopts the technical proposal that:

the utility model provides an utilize six orifice plates ring type transfection device, includes six orifice plates bodies, plate lid and electrode pairs, six orifice plates body intervals are equipped with six undercut's electrotransfection cell and hold the chamber, the electrode pair sets up in pairs every the electrotransfection cell holds the intracavity and is connected with the power through the wire, every electrode pair includes positive electrode and negative electrode, the positive electrode is the annular, the negative electrode is circular sheet structure, the negative electrode is located in the annular positive electrode, the cover can be dismantled and cover on six orifice plates bodies.

Further, the positive electrode is disposed along a circumferential edge of the electrotransfection cell-containing chamber.

Further, the negative electrode is located at the annular center of the positive electrode.

Further, the electrotransfected cell accommodating cavities are arranged in two rows and three columns.

Further, the lead wires are arranged along the edges and the center of the six-hole plate body and are respectively connected with each positive electrode and each negative electrode.

Further, the outer contour of the plate cover is larger than that of the six-hole plate body.

Further, the positive electrode and the negative electrode are both disposed at the bottom of the electrotransfection cell containing cavity.

Furthermore, the power supply is a pulse power supply, the pulse width of the power supply is 40-500us, and the long pulse is 10-50 ms.

Further, the high voltage of the power supply is 200-1000v, and the low voltage is 30-100 v.

Further, the high voltage of the power supply is 500-800v, and the low voltage is 50-80 v.

Additional aspects and advantages of the present application will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the present application.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the technical solutions in the prior art will be briefly described below. Throughout the drawings, like elements or portions are generally identified by like reference numerals. In the drawings, elements or portions are not necessarily drawn to scale.

FIG. 1 is a schematic diagram of a six-well plate circular transfection device provided in the examples of the present application;

FIG. 2 is a schematic diagram of a closed state of a six-well plate circular transfection device provided in the embodiments of the present application;

FIG. 3 is a front view of a circular transfection device utilizing a six-well plate as provided in the examples of the present application;

FIG. 4 is a side view of a circular transfection device utilizing a six-well plate as provided in the examples of the present application.

The battery comprises a positive electrode 1, a lead 2, a negative electrode 3, a six-hole plate body 4 and a plate cover 5.

Detailed Description

Here, it is to be noted that the functions, methods, and the like related to the present invention are only conventional adaptive applications of the related art. Therefore, the present invention is an improvement of the prior art, which substantially lies in the connection relationship between hardware, not in the functions and methods themselves, that is, the present invention relates to a few functions and methods, but does not include the improvements proposed in the functions and methods themselves. The present invention is described for better illustration of the function and method for better understanding of the present invention.

Embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and therefore are only examples, and the protection scope of the present invention is not limited thereby.

It is to be noted that unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the present invention belongs.

Referring to fig. 1 to 4, the present application provides an annular transfection device using a six-hole plate, including a six-hole plate 4, a plate cover 5 and electrode pairs, where the six-hole plate 4 is provided with six downward-recessed electrotransfection cell accommodating cavities at intervals, the electrode pairs are arranged in pairs in each of the electrotransfection cell accommodating cavities and connected to a power supply through a wire 2, each electrode pair includes a positive electrode 1 and a negative electrode 3, the positive electrode 1 is annular, the negative electrode 3 is a circular sheet structure, the negative electrode 3 is located in the annular positive electrode 1, and the plate cover 5 is detachably covered on the six-hole plate 4.

This application is through adopting six hole plate bodies 4 as the electrotransfection cell container, the interval sets up six undercut's electrotransfection cell and holds the chamber on this six hole plate bodies 4, it is connected with the power to set up mated electrode pair in every electrotransfection cell holds the intracavity, the device once can carry out the electrotransfection to six or six kinds of cells simultaneously, the electrotransfection is efficient, a large amount of sample repetitive operation's problem has been solved, and the survival rate of cell is high, transfection effect is good, has fine using value.

Electrotransfection cell holds the chamber and is the setting of multirow structure, in this embodiment, electrotransfection cell holds the chamber and is two rows of three columns settings, can carry out the electrotransfection to the sample that is located the electrotransfection cell and holds the intracavity simultaneously, guarantee that every electrotransfection cell holds the sample of intracavity and all obtains comparatively even electrotransfection, carry out the electrotransfection after the cavity is full of after the liquid gets into, the bubble that produces during the electrotransfection simultaneously also can be timely along with upper portion aperture discharge, after the electrotransfection is accomplished, liquid outflow cavity, the cavity forms atmosphere balance once more, the repeated formula feed liquor.

The positive electrode 1 is circular and arranged along the circumferential edge of the electrotransfection cell containing cavity, the negative electrode 3 is a flake and is positioned at the circular center of the positive electrode, and the positive electrode 1 and the negative electrode 3 are both positioned at the bottom of the electrotransfection cell containing cavity and are positioned in the same plane. By adopting the arrangement, the containing cavity can be completely filled with no liquid, and the electrotransfection can be carried out, so that the use is more convenient.

The distances between the positive electrodes 1 in the six electrotransfection cell containing cavities are the same, and the distances between the negative electrodes 3 are kept the same, so that the electrotransfection consistency of each containing cavity is ensured.

The lead wires 2 are disposed along the edges and the center of the six-hole plate body 4, and are connected to each of the positive electrodes 1 and the negative electrodes 3, respectively.

Specifically, an inward-recessed groove is formed along the edge and the center of the six-hole plate body 4, and the lead 2 is arranged along the groove and connected with the positive electrode 1 and the negative electrode 3 which are arranged in the corresponding electrotransfection cell accommodating cavities. A control switch is mounted on the lead 2.

The plate cover 5 is in a shape matched with the six-hole plate body 4, and the outer contour of the plate cover 5 is larger than that of the six-hole plate body 4, so that the plate cover 5 can be better covered on the six-hole plate body 4.

The power supply is a pulse power supply, the pulse width of the power supply is 40-500us, and the long pulse is 10-50 ms. The high voltage of the power supply is 200-1000v, and the optimal range of the high voltage is 500-800 v; the low voltage is 30-100v, and the optimal range of the low voltage is 50-80 v.

When electrotransfection is carried out, two modes can be adopted, namely high-voltage low-pulse-width tapping and low-voltage long-pulse inlet tapping, and the two modes are combined for use, so that the transfection efficiency can be obviously improved.

The cell transfection procedure using the device of the present application was as follows:

1. the cells and substances to be transfected are added to the electrotransfected cell receiving chamber in the six-well plate 4. The cells to be transfected are plant cells or animal cells, and can be stem cells, macrophages, lymphocytes and DC cells; the density of the cells to be transfected was 4X 10 ⁵ ～4×10 ⁷ one/mL. The substances to be transfected are siRNA, microRNA, DNA, proteins and polypeptides.

2. And starting a control switch to perform electrotransfection.

The electrotransfection device can perform large-volume transfection, and provides suitable application equipment for research and development of antibody drugs, immunotherapy, clinical diagnosis and the like.

In this application, unless expressly stated or limited otherwise, the terms "connected," "secured," and the like are to be construed broadly and can include, for example, fixed connections, removable connections, or integral combinations thereof; may be an electrical connection; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.

In the description of the present invention, a number of specific details are described. It is understood, however, that embodiments of the invention may be practiced without these specific details. In some instances, well-known methods, systems, and techniques have not been shown in detail in order not to obscure an understanding of this description.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, system, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, systems, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not substantially depart from the scope of the embodiments of the present invention, and are intended to be covered by the claims and the specification.

Claims (10)

1. The utility model provides an utilize six orifice plates ring type transfection device which characterized in that, includes six orifice plate bodies, plate lid and electrode pair, six orifice plate body intervals are equipped with six undercut's electrotransfection cell and hold the chamber, the electrode pair sets up in pairs every the electrotransfection cell holds the intracavity and is connected with the power through the wire, every electrode pair includes positive electrode and negative electrode, the positive electrode is the annular, the negative electrode is circular sheet structure, the negative electrode is located in the annular positive electrode, the cover can be dismantled and cover on six orifice plate bodies.

2. The device of claim 1, wherein the positive electrode is disposed along a circumferential edge of the electrotransfection cell-containing chamber.

3. The transfection device of claim 2, wherein the negative electrode is located in the center of the annulus of the positive electrode.

4. The device of claim 1, wherein the housing for the transfected cells is arranged in two rows and three columns.

5. The transfection device of claim 4, wherein the wires are disposed along the edges and center of the six-well plate body and are connected to each of the positive and negative electrodes, respectively.

6. The device of claim 1, wherein the outer profile of the plate cover is larger than the outer profile of the six-well plate body.

7. The device of claim 1, wherein the positive and negative electrodes are disposed at the bottom of the chamber.

8. The device of claim 1, wherein the power source is a pulsed power source with a pulse width of 40-500us and a long pulse of 10-50 ms.

9. The transfection device of claim 1, wherein the power supply has a high voltage of 200-1000v and a low voltage of 30-100 v.

10. The transfection device of claim 9, wherein the power supply has a high voltage of 500-800v and a low voltage of 50-80 v.

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