JPS63141575A - Electrode of cell electric stimulating apparatus - Google Patents
Electrode of cell electric stimulating apparatusInfo
- Publication number
- JPS63141575A JPS63141575A JP61289954A JP28995486A JPS63141575A JP S63141575 A JPS63141575 A JP S63141575A JP 61289954 A JP61289954 A JP 61289954A JP 28995486 A JP28995486 A JP 28995486A JP S63141575 A JPS63141575 A JP S63141575A
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- separator
- electrode plates
- thin electrode
- electrode sheets
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 230000004936 stimulating effect Effects 0.000 title 1
- 230000000638 stimulation Effects 0.000 claims description 6
- 230000004927 fusion Effects 0.000 abstract description 6
- 239000006285 cell suspension Substances 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 10
- 239000002184 metal Substances 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000000034 method Methods 0.000 description 4
- 230000007910 cell fusion Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000004925 Acrylic resin Substances 0.000 description 2
- 229920000178 Acrylic resin Polymers 0.000 description 2
- 230000005684 electric field Effects 0.000 description 2
- 229910001369 Brass Inorganic materials 0.000 description 1
- 239000010951 brass Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は細胞膜のみを有する細胞どおしを電気的に融合
させたり、細胞に遺伝子を導入させたりする細胞電気刺
激装置に使用される電極に関するものである。Detailed Description of the Invention (Industrial Application Field) The present invention relates to an electrode used in a cell electrical stimulation device that electrically fuses cells having only cell membranes or introduces genes into cells. It is related to.
（従来の技術）
細胞電気刺激装置では細胞懸濁液を収容する容器と電極
とが一体化した構造の細胞電気刺激チャンバを使用する
のが一般的である。しがし、電極と容器が一体化された
細胞電気刺激チャンバでは、細胞懸濁液の処理量に限界
がある。(Prior Art) A cell electrical stimulation device generally uses a cell electrical stimulation chamber having a structure in which a container containing a cell suspension and an electrode are integrated. However, in a cell electrical stimulation chamber in which an electrode and a container are integrated, there is a limit to the amount of cell suspension that can be processed.
そこで、大量の細胞懸濁液を処理する方法として、電極
を容器から分離し、Ｉａ胞懸濁液を収容する容器を多数
配置し、それらの容器に電極を順次挿入していく方法が
提案されている（「バイオサイエンス畷レポート（Ｂｉ
ｏｓｃｉｅｎｃｅ Ｒｅｐｏｒｔ、ｓ） Ｊ誌、第４巻
、３３５〜３４２ページ（１９８４年）参照）。Therefore, as a method for processing a large amount of cell suspension, a method has been proposed in which the electrode is separated from the container, a number of containers containing the Ia cell suspension are arranged, and the electrodes are sequentially inserted into these containers. (“Bioscience Nawate Report (Bi)
Science Report, s) J, Vol. 4, pp. 335-342 (1984)).
上記引用例で提案された電極を第４図に示す。The electrode proposed in the cited example above is shown in FIG.
電極は複数の薄板電極板２をアクリル樹脂製のセパレー
タによって互いに平行に、がっ、電極板２間の間隔が一
定になるように保持されて固定されている。この電極板
２の集合体の水平方向の平面形状の輪郭は正方形であり
、したがって細胞懸濁液を収容し、この電極を浸す容器
４は角型容器である。The electrodes are fixed by holding a plurality of thin electrode plates 2 in parallel with each other by separators made of acrylic resin so that the spacing between the electrode plates 2 is constant. The horizontal planar outline of this assembly of electrode plates 2 is square, and therefore the container 4 that accommodates the cell suspension and in which this electrode is immersed is a square container.
（発明が解決しようとする問題点）
細胞培養の領域では角型容器は入手が困難である。上記
の方法では角型の容器を用いているために普及しなかっ
た。(Problems to be Solved by the Invention) Square containers are difficult to obtain in the field of cell culture. The above method did not become popular because it used a square container.
容器の普及形状は第５図に記号６で示されるように円形
である１円形容器６に合致する電極は記号８で示される
ような同心円状のものが好ましい。The general shape of the container is circular as shown by symbol 6 in FIG. 5. The electrodes matching the circular container 6 are preferably concentric circles as shown by symbol 8.
しかし、同心円状の電極８では、電界強度にｒ １／
ｒ ２なる差異が生じるために融合や遺伝子導入の効率
を定量的に把握することができない問題がある。However, in the concentric electrode 8, the electric field strength is r 1/
There is a problem in that the efficiency of fusion and gene transfer cannot be quantitatively understood because of the r2 difference.
本発明は、均一な電界を発生することができ、平面形状
の輪郭が円形であって円形の容器に使用することができ
、更に融合や遺伝子導入の過程を顕微鏡でｗｔ察するこ
とのできる電極を提供することを目的とするものである
。The present invention provides an electrode that can generate a uniform electric field, has a circular planar outline, can be used in a circular container, and can further observe the process of fusion and gene transfer with a microscope. The purpose is to provide
（問題点を解決するための手段）
本発明の電極では、複数の薄板電極板を上端部で１枚お
きに電気的接続を行なって束ねてそれぞれの束に端子を
設け、これらの複数の薄板電極板の下端部においては下
端から所定の高さだけ上った位置で透明の絶縁性セパレ
ータを薄板電極板間に介在させて、薄板電極板が互いに
平行に、かつ、一定間隔を保った状態に全ての薄板電極
板を束ね。(Means for Solving the Problems) In the electrode of the present invention, a plurality of thin electrode plates are electrically connected to each other at the upper end, and a terminal is provided in each bundle. At the lower end of the electrode plate, a transparent insulating separator is interposed between the thin electrode plates at a predetermined height above the lower end, so that the thin electrode plates are parallel to each other and maintained at a constant interval. Bundle all the thin electrode plates together.
この薄板電極板の集合体の平面形状の輪郭が円形になる
ように前記各薄板電極板及び前記セパレータの幅を定め
た。The widths of each of the thin electrode plates and the separator were determined so that the outline of the planar shape of the assembly of thin electrode plates was circular.
（実施例）
第１図は一実施例を表わす平面図、第２図は第１図のＡ
−Ｂ線位置での断面図、第３図は第１図のＣ−Ｄ線位置
での断面図である。(Example) Figure 1 is a plan view showing one example, and Figure 2 is A of Figure 1.
3 is a sectional view taken along line C--D in FIG. 1. FIG.
１０−１．１０−２．・・・・・・は薄板電極板であり
。10-1.10-2. ... is a thin electrode plate.
例えば厚さが０．１ｍｍの真鍮板である。各薄板電極板
１０−１，１０７２．・・・・・・は第３図に示される
ように、下部の電極板本体１２ａと上部の端子接続部１
２ｂとからなっている。端子接続部１２ｂは奇数番目の
薄板電極板ｔｏ−１，１０−３゜・・・・・・において
は第３図で中心より左側方向に突出し、偶数番目の薄板
電極板１０−２．１０−４゜・・・・・・においては第
３図で中心より右側方向に突出している。奇数番目の薄
板電極板１０−１．１０−３．・・・・・・の端子接続
部１２ｂは金属製のカラー１４を介して束ねられ、端子
接続部１２ｂに設けられた孔に端子１６と一体的に形成
された金属棒が通され、その金属棒の終端がナツトで絞
めつけられることによって固定されている。偶数番目の
薄板電極板１０−２．１０−４．・・・・・・について
も同様にして、それらの端子接続部１２ｂが金属製のカ
ラー１４を介して束ねられ、端子接続部１２ｂに設けら
れた孔に端子１７と一体的に形成された金属棒が通され
、その金属棒の終端がナツトで絞めつけられることによ
って固定されている。For example, it is a brass plate with a thickness of 0.1 mm. Each thin electrode plate 10-1, 1072. . . . as shown in FIG. 3, the lower electrode plate main body 12a and the upper terminal connection portion
It consists of 2b. The terminal connection portion 12b protrudes to the left side from the center in FIG. At 4°..., it protrudes to the right from the center in Fig. 3. Odd numbered thin plate electrode plate 10-1.10-3. The terminal connecting portions 12b are bundled together via a metal collar 14, and a metal rod integrally formed with the terminal 16 is passed through a hole provided in the terminal connecting portion 12b. The end of the rod is secured by tightening with a nut. Even-numbered thin plate electrode plates 10-2.10-4. . . . Similarly, the terminal connecting portions 12b are bundled together via the metal collar 14, and the metal terminals 17 are integrally formed in the holes provided in the terminal connecting portions 12b. A rod is passed through it, and the end of the metal rod is secured by tightening with a nut.
薄板電極板１０−１，１０−２．・・・・・・の電極板
本体１２ａにおいては、２個の孔２０．２０があけられ
、薄板電極板１０−１．１０−２．・・・・・・の間に
アクリル樹脂の如き透明樹脂製の絶縁性セパレータ１８
を介在させて各薄板電極板１０−１゜１０−２．・・・
・・・の間隔が一定となるようにし、薄板電極板１０−
１．１０−２． ・・・・・の孔２０，２０とセパレ
ータ１８，１８．・・・・・・の孔にボルト２１．２１
を通してナツト２２．２２で絞めつけることによって、
薄板電極板１０−１．１０−２゜・・・・・・が互いに
平行に、かつ、一定間隔を保った状態で固定されている
。セパレータ１８の位置は。Thin electrode plates 10-1, 10-2. In the electrode plate main body 12a of . An insulating separator 18 made of transparent resin such as acrylic resin is placed between...
Each thin electrode plate 10-1°10-2. ...
... so that the intervals between the thin electrode plates 10-
1.10-2. ... holes 20, 20 and separators 18, 18. Bolt 21.21 in the hole of...
By tightening with a nut 22.22 through the
Thin electrode plates 10-1, 10-2°, . . . are fixed in parallel with each other and at constant intervals. What is the position of the separator 18?
薄板電極板１０−１．１０−２．・・・・・・の下端部
が細胞に電圧を印加できるように、薄板電極板１０−１
．１０−２．・・・・・・の下端から所定の高さだけ上
がった位置に設定されている。Thin electrode plate 10-1.10-2. Thin electrode plate 10-1 so that the lower end can apply voltage to the cells
．． 10-2. It is set at a position that is a predetermined height above the bottom edge of...
薄板電極板１０−１．１０−２．・・・・・・とセパレ
ータ１８，１８．・・・・・・の集合体としての電極の
平面形状は、第１図に示されるようにその輪郭がほぼ円
形になるように薄板電極板１０−１．ｔｏ−２゜・・・
・・・とセパレータ１８，１８．・・・・・・の幅（第
３図中のＷ）が定められている。Thin electrode plate 10-1.10-2. ...and separators 18, 18. The planar shape of the electrodes as an assembly of thin electrode plates 10-1, . to-2゜...
... and separators 18, 18. . . . width (W in FIG. 3) is determined.
セパレータ１８，１８．・・・・・・には中央に排気空
間用の孔２４が開けられている。Separators 18, 18. . . . has a hole 24 for an exhaust space in the center.
２６は細胞懸濁液を収容し、電極を浸して細胞に電気刺
激を与えて融合や遺伝子導入をさせるための容器として
のウェル又はペトリシャーレである。Reference numeral 26 designates a well or petri dish that serves as a container for containing a cell suspension and immersing an electrode therein to apply electrical stimulation to the cells to cause fusion or gene transfer.
次に、本実施例の電極を細胞融合に使用する場合の動作
について説明する。Next, the operation when using the electrode of this example for cell fusion will be explained.
ウェル２６の底に細胞懸濁液２８を所定量ずつ収容して
おく。ウェル２６に電極を入れると、セパレータ１８の
下面と細胞懸濁液２８の間にある空気が孔２４と、セパ
レータ１８とウェル２６の器壁との間の空間を通って上
方へ逃げる。このことで細胞懸濁液の液面の揺れが防止
され、細胞が破壊されるのが抑えられる。A predetermined amount of the cell suspension 28 is stored at the bottom of the well 26. When the electrode is placed in the well 26, air between the lower surface of the separator 18 and the cell suspension 28 escapes upward through the hole 24 and the space between the separator 18 and the wall of the well 26. This prevents the liquid level of the cell suspension from shaking and suppresses cell destruction.
端子１６と端子１７の間に電圧を印加し、細胞を電気的
に融合させる。このとき、光源からの光が孔２４を通る
ようにして細胞融合状態を顕微鏡でｌ！察すると、セパ
レータ１８直下から鮮明な顕微１Ｉｔａｌ）を行なうこ
とができる。A voltage is applied between terminals 16 and 17 to electrically fuse the cells. At this time, light from the light source passes through the hole 24 and the state of cell fusion is observed using a microscope. As you can see, clear microscopy can be performed from directly below the separator 18.
所定の融合操作が終了すると、次のウェル２６にこの電
極を入れて同様に細胞融合操作を繰り返していく。When the predetermined fusion operation is completed, this electrode is placed in the next well 26 and the cell fusion operation is repeated in the same manner.
なお、細胞懸濁液２８は予めウェル２６に入れておいて
もよく、また、電極をウェル２６に入れた後に細胞懸濁
液２８を注入してもよい。Note that the cell suspension 28 may be placed in the well 26 in advance, or may be injected after the electrode is placed in the well 26.
上記の実施例では、セパレータ１８に孔２４を設けてい
るので、薄板電極板１０−１．１０−２゜・・・・・・
とセパレータ１８，１８．・・・・・・で囲まれた細胞
懸濁液２８の液面をあまり揺らすことなく電極をウェル
２６から出入れでき、壊れやすい細胞を液の揺れで壊れ
るのを防止することができる。特に、細胞懸濁液２８の
液面がセパレータ１８の下面に接する場合に有効である
。しかし、本発明は、セパレータ１８に孔２４が設けら
れていない場合も含むものである。In the above embodiment, since the holes 24 are provided in the separator 18, the thin electrode plates 10-1, 10-2 degrees...
and separators 18, 18. The electrode can be taken in and out of the well 26 without shaking the liquid surface of the cell suspension 28 surrounded by . This is particularly effective when the liquid surface of the cell suspension 28 comes into contact with the lower surface of the separator 18. However, the present invention also includes a case where the separator 18 is not provided with the holes 24.
（発明の効果）
（１）本発明の電極では、薄板電極板を互いに平行に、
かつ、一定間隔を保った状態に維持するセパレータとし
て透明なセパレータを使用したので、倒立型顕微鏡で融
合状態や遺伝子導入状態を常時ａ察することができる。(Effects of the invention) (1) In the electrode of the present invention, the thin electrode plates are arranged parallel to each other.
In addition, since a transparent separator was used to maintain a constant spacing, the fusion state and gene transfer state can be observed at all times using an inverted microscope.
（２）平面形状の輪郭を円形にしたので、入手の容易な
円形の容器を使用することができる。(2) Since the outline of the planar shape is circular, easily available circular containers can be used.
（３）ｇ板電極板を使用しているので、容量を大きくす
ることができ、細胞懸濁液の処理量を大きくすることが
できる。(3) Since the G-plate electrode plate is used, the capacity can be increased, and the amount of cell suspension that can be processed can be increased.
（４）小型に作ることができるので、顕微鏡下の狭いス
ペースに設置することができる。(4) Since it can be made small, it can be installed in a narrow space under a microscope.
第１図は一実施例を容器とともに示す平面図、第２図は
第１図のＡ−Ｂ１位置での断面図、第３図は第１図のＣ
−Ｄ線位置での断面図、第４図は従来の電極と容器を示
す概略斜視図、第５図は他の従来の電極と容器を示す概
略斜視図である。
ｔｏ−ｔ、ｔｏ−２，・・・・・・；薄板電極板。
１６．１７；端子、
１８；セパレータ。Figure 1 is a plan view showing one embodiment together with a container, Figure 2 is a sectional view taken along line A-B1 in Figure 1, and Figure 3 is C in Figure 1.
4 is a schematic perspective view showing a conventional electrode and container, and FIG. 5 is a schematic perspective view showing another conventional electrode and container. to-t, to-2,...; thin electrode plate. 16.17; Terminal; 18; Separator.
Claims (1)
接続を行なって束ねてそれぞれの束に端子を設け、これ
らの複数の薄板電極板の下端部においては下端から所定
の高さだけ上った位置で透明の絶縁性セパレータを薄板
電極板間に介在させて、薄板電極板が互いに平行に、か
つ、一定間隔を保った状態に全ての薄板電極板を束ね、
この薄板電極板の集合体の平面形状の輪郭が円形になる
ように前記各薄板電極板及び前記セパレータの幅を定め
た細胞電気刺激装置の電極。(1) A plurality of thin electrode plates are electrically connected every other sheet at the upper end, and a terminal is provided in each bundle, and the lower end of the plurality of thin electrode plates is set at a predetermined height from the lower end. At the upper position, a transparent insulating separator is interposed between the thin electrode plates, and all the thin electrode plates are bundled so that the thin electrode plates are parallel to each other and are kept at a constant interval.
An electrode for a cell electrical stimulation device, wherein the width of each of the thin electrode plates and the separator is determined so that the outline of the planar shape of the aggregate of the thin electrode plates is circular.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61289954A JPH0695926B2 (en) | 1986-12-04 | 1986-12-04 | Electrodes of cell electrical stimulator |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61289954A JPH0695926B2 (en) | 1986-12-04 | 1986-12-04 | Electrodes of cell electrical stimulator |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63141575A true JPS63141575A (en) | 1988-06-14 |
| JPH0695926B2 JPH0695926B2 (en) | 1994-11-30 |
Family
ID=17749883
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61289954A Expired - Fee Related JPH0695926B2 (en) | 1986-12-04 | 1986-12-04 | Electrodes of cell electrical stimulator |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0695926B2 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH03143381A (en) * | 1989-10-27 | 1991-06-18 | Kurabo Ind Ltd | Electrode for electrical cell operation |
| JP2009043661A (en) * | 2007-08-10 | 2009-02-26 | Furukawa Electric Co Ltd:The | Multipolar connector and its assembling method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS58164500U (en) * | 1982-04-30 | 1983-11-01 | 植田 茂樹 | Stamp stand for replica of bacterial colony |
-
1986
- 1986-12-04 JP JP61289954A patent/JPH0695926B2/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS58164500U (en) * | 1982-04-30 | 1983-11-01 | 植田 茂樹 | Stamp stand for replica of bacterial colony |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH03143381A (en) * | 1989-10-27 | 1991-06-18 | Kurabo Ind Ltd | Electrode for electrical cell operation |
| JP2009043661A (en) * | 2007-08-10 | 2009-02-26 | Furukawa Electric Co Ltd:The | Multipolar connector and its assembling method |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0695926B2 (en) | 1994-11-30 |
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