JPS63276478A - Chamber for giving electric stimulation to cells - Google Patents

Abstract

PURPOSE:To enable cell fusion of high efficiency, by combining the first and second multi-ring electrodes of integral structures having a plurality of annular electrode plates, to arrange the electrodes alternately, as insulators are laid between the electrodes, so that the intervals and parallelism between electrode plates are made more precise. CONSTITUTION:In the vessel 1, the first multi-ring electrode 3 lathed into an integral structure (the lower electrode) is placed on the bottom. An insulator 5 is extended on the bottom surface of the electrode 3, and cell suspension 7 is fed on the insulator 5. Then, the second multi-ring electrode 4 (upper electrode) which have plate diameters different from those of the first electrode 3 and is provided with an electric terminal 6 on its cover is combined with the lower electrode 3, as an insulator 18 is laid between them. At this time, the terminal rode 14 of the lower electrode 3 is extended upward through the electrode 4 and the hole of the cover 2. A prescribed voltage is loaded between the terminals 14 and 6 to effect fusion of the cells in the suspension 7.

Description

[Detailed Description of the Invention] (Industrial Application Field) The present invention relates to a cell electrical stimulation chamber used to fuse cells or introduce genes into cells using electrical stimulation. It is.

(Prior art) In a cell electrical stimulation chamber, a pair of flat plate electrodes are generally provided parallel to each other, and cells are placed between the pair of electrodes)!
! A turbid cell suspension is placed in the cell, and a voltage is applied between the electrodes to provide electrical stimulation to the cells, causing the cells to fuse or introducing genes into the cells.

If a large amount of cell suspension is to be processed, a structure must be provided with a large number of pairs of parallel plate ffi poles. As a cell electrical stimulation chamber for mass processing, there is one in which a plurality of such parallel plate electrodes are stacked on top of each other at equal intervals with insulating spacers interposed therebetween.

(Problems to be Solved by the Invention) In a cell electrical stimulation chamber, the conditions for electrical stimulation applied to cells change depending on the spacing and inclination between the electrodes, so it is necessary to keep a plurality of flat electrodes at a constant spacing from each other. Moreover, it is not easy to assemble the electrodes while maintaining parallelism, and therefore, there is a problem in that the cost of such electrodes for mass processing is high.

An object of the present invention is to provide a cell electrical stimulation chamber for mass processing that can be manufactured at a low cost while maintaining mechanical precision such as the spacing and parallelism between electrode plates.

(Means for solving the problem) To explain with reference to FIG. 1 and FIG. 2 showing an example,
The cell electrical stimulation chamber of the present invention includes a first multi-ring electrode (lO) fabricated in one piece with a plurality of concentric electrode plates (lO).
3), and a second multi-ring electrode (3) integrally formed with a plurality of concentric electrode plates (16) having different radii.
using a multi-ring electrode (4), with the bottom (12) of the first multi-ring electrode (3) and the second multi-ring electrode (4)
An insulator (5) is interposed between the electrode plate (16) of the first multi-ring electrode (3) and the second multi-ring electrode (4) between the electrode plate (10) of the first multi-ring electrode (3). This is the combination with the plate (16) inserted.

(Example) FIG. 1 is a sectional view showing one example, and FIG. 2 is a plan view. Note that FIG. 1 is a sectional view taken along line A-A in FIG. 2.

1 is a container made of transparent resin, and for example, a Petri dish can be used. A lower electrode 3, which is a first multi-ring electrode, is placed in the container 1 with its electrode plate IO facing upward. In the lower electrode 3, a plurality of concentric electrode plates 10 are integrally provided on a bottom plate 12 in a direction perpendicular to each other, and a terminal bar 14 is integrally provided in the center. Lower electrode 3
is manufactured by lathe processing.

2 is a lid made of transparent resin, and an upper electrode 4, which is a second multi-ring electrode, is fixed to the lid 2 so that its electrode plate 16 faces downward as shown in the figure. In order to fix the upper electrode 4 to the lid 2, a screw hole is drilled in the bottom of the upper electrode 4, a hole is punched in the lid 2, and a terminal 6 is screwed into the screw hole of the upper electrode 4 from the outside of the lid 2. ing. The upper electrode 4 is also manufactured by lathe processing, and has a plurality of concentric electrode plates 1 on the bottom plate.
6 are integrally provided in the orthogonal direction.

A hole is made in the center of the upper electrode 4 and the lid 2, and the lid 2 is connected to the container 1.
When combining the upper electrode 4 and the lower electrode 3 by placing them over the upper electrode 3, the terminal rod 14 of the lower electrode 3 is passed through the hole, and the relative positioning of the lower electrode 3 and the upper electrode 4 is performed. Terminal bar 14
An insulator 18 is interposed between the upper electrode 4 and the upper electrode 4 to maintain insulation between the upper electrode 4 and the lower electrode 3.

An elastic transparent insulator 5 is laid on the inner bottom of the lower electrode 3, and when the lower electrode 3 and the upper electrode 4 are combined, the lower electrode 3
The bottom of the upper electrode 4 and the tip of the electrode plate 16 of the upper electrode 4 are insulated.

The electrode plates 10 of the lower electrode 3 are formed in concentric circles at equal intervals, and the electrode plates 16 of the upper electrode 4 are also formed in concentric circles at equal intervals.

The electrode plate IO of the lower electrode 3 and the electrode plate 16 of the upper electrode 4 are set so that the radius of one electrode plate 10 (or 16) is in the middle of the radius of the other adjacent electrode plate 16 (or IO). do. Thereby, when both the electrodes 3 and 4 are combined as shown in FIG. 1, the plurality of intervals between the electrode plates 10 and the electrode plates 16 become equal.

The bottoms of the lower electrode 3 and the upper electrode 4 are each partially provided with an opening 20, 22 as shown in FIG.
When both electrodes 3 and 4 are combined, opening 2 of both electrodes 3 and 4
0.22 overlap each other, allowing light to pass from above the lid 2 to below the container 1. Therefore, by placing the objective lens of an inverted microscope below the container l, it is possible to observe the process of cell fusion and gene transfer within this cell electrical stimulation chamber using a microscope.

Next, the operation of this embodiment will be explained.

A lower electrode 3 is placed in a container 1 in the state shown in the figure, an insulator 5 is placed on the bottom of the lower electrode 3, and a cell suspension 7 is injected onto the insulator 5.

With the upper electrode 4 attached to the lid 2 by the terminal 6 as shown in the figure, pass the terminal rod 14 through the hole in the upper electrode 4 and the lid 2 via the insulator 18, and combine the upper electrode 4 with the lower electrode 3. .

A predetermined voltage is applied to the terminal bar 14 and the terminal 6 to remove the cell suspension 7.
fuse the cells and the oshi.

The process of cell fusion can be observed under the microscope through opening 20.22.
I guess.

(Effects of the Invention) In the present invention, both electrodes to be combined can be manufactured by lathe processing, so the processing cost is lower than the conventional method of assembling flat plate electrodes while maintaining mechanical precision.

Since each electrode plate is integrally processed by lathe processing, mechanical precision such as spacing and parallelism between them is high.

In the case of electrodes made by assembling conventional flat plate electrodes, the tips of the electrodes are uneven, resulting in a gap between the electrode tips and the bottom surface.

In cell fusion, electrical stimulation is applied to the cells precipitated on the bottom surface to induce fusion, which has the disadvantage that the S1 merging efficiency is poor.

In contrast, in the present invention, since the electrical bridge is lathed and integrated, the tip surfaces of the electrode plates will not be uneven, which is effective in increasing the efficiency of cell fusion.

[Brief explanation of drawings]

FIG. 1 is a cross-sectional view showing one embodiment, and FIG. 2 is a plan view of the same embodiment, showing a cut state taken along the line A--A in FIG. 1... Container, 2... Lid, 3... Lower electrode, 4... Upper electrode, 5... Insulator, 7. ...Cell suspension, 10.16... Electrode plate.

Claims (4)

[Claims] (1) A first unit that is integrally processed with multiple concentric electrode plates
A multi-ring electrode is housed in a container, and a second multi-ring electrode, which is integrally processed with the first multi-ring electrode and has a plurality of concentric electrode plates having different radii, is fixed to the lid. An insulator is interposed between the bottom of the electrode and the tip of the electrode plate of the second multiple ring electrode, and the electrode plate of the second multiple ring electrode is inserted between the electrode plates of the first multiple ring electrode. Combined cell electrical stimulation chamber. (2) The container, the lid, and the insulator are made of a transparent material, and openings are partially provided at the bottom of the first multiple ring electrode and the bottom of the second multiple ring electrode, and both multiple ring electrodes are provided with openings. The cell electrical stimulation chamber according to claim 1, wherein the positions of these openings of the electrodes are overlapped to allow light to pass therethrough. (3) The cell electrical stimulation chamber according to claim 1, wherein the insulator is made of an elastic material. (4) A terminal bar is integrally machined in the center of the first multiple ring electrode, a hole is made in the center of the second multiple ring electrode and the center of the lid, and the terminal bar is provided with a hole in the center of the second multiple ring electrode. The second multiple ring electrode is positioned with respect to the first multiple ring electrode by passing through the second multiple ring electrode and the hole in the lid while being insulated from the second multiple ring electrode. Cell electrical stimulation chamber according to scope 1.