JPH058876Y2 - - Google Patents

Description

[Detailed Description of the Invention] (Industrial Application Field) The present invention relates to a cell fusion chamber that provides a special space for fusion of cells in a cell fusion device that electrically fuses cells.

(Prior Art) As an example of a cell fusion chamber, one shown in FIG. 2 has been proposed.

2 is a chamber body, which is formed into a dish shape. A lid 4 is adapted to be placed over the upper opening of the chamber body 2 in an airtight manner. A pair of electrodes 6 and 8 are assembled on the bottom plate 2a within the chamber body 2, facing each other, and between these pair of electrodes 6 and 8, a cell suspension 1 containing cells is suspended.
0 is now accommodated. The electrodes 6 and 8 are provided with lead wires 12 and 14, respectively, and these lead wires 12 and 14 are taken out to the outside through the side wall 2b of the chamber body.

Cell fusion operations are often performed while observing under a microscope. Therefore, light is irradiated from above the lid 4, and the cell suspension 10 and the bottom plate 2b of the chamber body are
The image is then observed with an inverted microscope through the lens system 16 below. Therefore, the chamber body 2 and the lid 4 are made of transparent materials.

Conventionally, transparent resin such as polycarbonate has been used for the chamber body 2 and the lid 4, for example.

(Problems to be Solved by the Invention) In the conventional cell fusion chamber, the chamber body 2 is made of resin, so there are the following problems.

The operation of introducing and removing the cell suspension 10 into the space between the electrodes 6 and 8 is usually performed using a glass pipette or Pasteur.
At that time, the tip of a pipette or the like may come into contact with the surface of the bottom plate 2a of the chamber body. Since resin is softer than glass, it is easily scratched. If the surface of the bottom plate 2a of the chamber body is scratched, the transparency will deteriorate, causing problems in microscopic observation. There is also the problem that cells fall into the scratched areas on the surface of the bottom plate 2a, restricting cell movement for fusion, and making fusion difficult.

Therefore, for example, it is possible to use a glass shear plate as the chamber body 2 and assemble the electrodes 6 and 8 on the bottom plate thereof, but it is also possible to assemble the electrodes 6 and 8 on the bottom plate.
In order to take out No. 4 to the outside, a hole must be made in the side wall of the shear tray. However, such hole machining is not easy.

The purpose of the present invention is to provide a cell fusion chamber that is hard to damage the bottom plate of the chamber body, allows smooth cell fusion, and allows arbitrary observation under a microscope. .

(Means for Solving the Problems) In the cell fusion chamber of the present invention, a resin side wall is bonded with adhesive to a transparent glass plate serving as a bottom plate to form a dish-shaped chamber body. The opening is covered with a transparent lid to keep it airtight, and a pair of electrodes are assembled on the bottom glass plate in the chamber body so as to face each other, and a cell suspension is placed between the pair of electrodes. The lead terminals from each of the electrodes are taken out to the outside through the resin side wall.

(Example) Figure 1 shows an embodiment of the present invention, Figure A is a plan view showing the inside with a part of the lid cut away, and Figure B is a plan view with part of the lid and chamber body cut away. FIG. 3 is a front view showing the inside.

Reference numeral 20 denotes a transparent glass plate that serves as the bottom plate of the chamber body. On the transparent glass plate 20, a ring-shaped inner electrode 22 and a ring-shaped outer electrode 24 having a larger diameter than the ring-shaped inner electrode 22 are arranged parallel to each other. Adhesive 26,
28 and assembled. The inner electrode 22 and the outer electrode 24 are concentric, and both electrodes 2
The gaps 2 and 24 serve as a space for accommodating the cell suspension.

30 is a side wall of the chamber body, which is made of resin. An adhesive 3 is applied between the side wall 30 and the glass plate 20 and between the side wall 30 and the side surface of the outer electrode 24.
2.

The adhesives 26, 28, and 32 do not change in quality even when the entire chamber is heated in an autoclave and sterilized with steam, and the adhesive does not peel off due to the difference in temperature expansion coefficient of each part when heated. It is necessary that the substance does not produce any harmful substances, and at the same time it must be harmless and non-toxic to living things. As such an adhesive, for example, Dow Corning's silicone adhesive DC SILASTIC 739 WHITE can be used.

A lead terminal 34 for the inner electrode 22 is attached to the side wall 30 of the chamber body in an airtight manner with sheet packings 38a, 38b, and a lead terminal 36 for the outer electrode 24 is attached in an airtight manner with sheet packings 40a, 40b. There is. A washer 42 is attached to the chamber of the inner electrode lead terminal 34 with a nut, a washer 44 is attached to the inner electrode 22 with a bolt, and the washers 42 and 44 are connected by an adhesive terminal 46.
Similarly, a washer 48 is attached to the chamber of the lead terminal 36 for the outer electrode with a nut.
Further, a washer 50 is attached to the outer electrode 24 with a bolt, and the washers 48 and 50 are connected by a connecting terminal 52.

The upper part of the chamber body consisting of the glass plate 20 and the side wall 30 is open, and this upper opening is covered with a lid 54 made of transparent resin. The lid 54 and the upper end surface of the side wall 30 of the chamber main body are in close contact with each other to maintain airtightness. In this way, the airtightness between the chamber body and the lid is maintained, and
The lead terminal 3 is connected to the side wall 30 of the chamber body.
4 and 36 are attached in an airtight manner with sheet packing, the inside of the chamber main body can be kept in a sealed state, and outside air can be shut off to ensure a sterile state.

The inner circular portion of the ring-shaped inner electrode 22 serves as a liquid reservoir, and by placing water therein, it is possible to prevent the cell suspension from evaporating. The airtight structure between the chamber body and the lid 54 and the airtight structure of the lead electrodes can prevent water introduced into the inner electrode 22 from evaporating.

To perform cell fusion using the cell fusion chamber of the present invention, a cell suspension is poured into the gap between the inner electrode 22 and the outer electrode 24, water is poured into the inner part of the inner electrode 22, and the lid 54 is closed. In that state, lead terminal 3
A voltage is applied to the inner electrode 22 and the outer electrode 24 from 4 and 36, and cell fusion is caused by a conventional method.
The process of cell fusion can be observed by irradiating light from above the lid 54 and passing through the cell suspension using an inverted microscope below the glass plate 20.

Although the embodiment shown in FIG. 1 is a circular chamber, parallel plate electrodes may be used instead of the pair of ring-shaped electrodes. Even in that case, the same effect can be obtained by using a glass plate as the bottom plate, a resin as the side wall, and bonding them together with an adhesive.

(Effects of the invention) In the cell fusion chamber of the invention, the side walls of the chamber body are made of resin and the bottom plate is a transparent glass plate.
Since the two are bonded with adhesive, the bottom plate will not be damaged even during long-term use of this cell fusion chamber, making it possible to observe cells with a microscope for a long period of time.

In addition, since the bottom plate had a smooth glass surface, the movement of cells during cell fusion was improved, resulting in a high fusion rate.

The cell fusion chamber of the present invention can be manufactured at a lower cost than when the bottom plate and side walls of the chamber body of a conventional cell fusion chamber are made of resin, and the resin is machined to make the bottom plate flat. be able to.

[Brief explanation of drawings]

Fig. 1 is a diagram showing an embodiment of the present invention, and Fig. 1A is a plan view showing the inside with a part of the lid cut away;
FIG. 2B is a front view showing the interior of the cell fusion chamber with a portion of the lid and side wall of the chamber main body cut away, and FIG. 2 is a sectional view showing the conventional cell fusion chamber. 20...Bottom glass plate, 22...Inner electrode, 24
...Outer electrode, 30...Chamber main body side wall, 34,
36...Lead terminal, 54...Lid.

Claims (1)

[Claim for Utility Model Registration] A resin side wall is glued to a transparent glass plate serving as the bottom plate to form a dish-shaped chamber body, and the upper opening of the chamber body is provided with a transparent lid to maintain airtightness. A pair of electrodes are assembled to face each other on the bottom glass plate within the chamber main body, and a cell suspension is accommodated between the pair of electrodes, and a cell suspension is accommodated between the pair of electrodes. A cell fusion chamber in which lead terminals from each electrode are taken out to the outside through the resin side wall.