JP2590459B2 - Cell capture device - Google Patents
Cell capture deviceInfo
- Publication number
- JP2590459B2 JP2590459B2 JP19581886A JP19581886A JP2590459B2 JP 2590459 B2 JP2590459 B2 JP 2590459B2 JP 19581886 A JP19581886 A JP 19581886A JP 19581886 A JP19581886 A JP 19581886A JP 2590459 B2 JP2590459 B2 JP 2590459B2
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- cells
- chamber
- capillary
- cell
- 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.)
- Expired - Fee Related
Links
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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Description
【発明の詳細な説明】 （イ） 産業上の利用分野 本発明は、細胞懸濁液中の細胞を捕獲する装置に関す
る。The present invention relates to an apparatus for capturing cells in a cell suspension.
（ロ） 従来技術 細胞懸濁液中の細胞に機械的にDNAを入れたり、ある
いは細胞内の物質を抽出する際には懸濁液中の細胞を捕
獲固定する必要がある。(B) Prior art When mechanically inserting DNA into cells in a cell suspension or extracting a substance in the cells, it is necessary to capture and fix the cells in the suspension.
この必要性に対処するため従来では、細胞よりやや小
さ目なガラスキャピラリを用い、それを負圧して細胞を
捕獲固定していた。Conventionally, to address this need, glass capillaries that are slightly smaller than cells have been used, and negative pressure has been applied to capture and fix the cells.
（ハ） 発明が解決しようとする問題点 ガラスキャピラリを使用する場合、液体中に浮遊して
いる細胞を自由に傷つけずに捕獲することが大変困難で
あった。なぜなら浮遊中の細胞はガラスキャピラリを近
づけると、それによって起こる水流で逃避しまた、負圧
度を大きくして遠方位から吸着しようとするとその吸着
力が細胞をこわす原因となるからである。これは特に小
さな細胞ほど顕著であった。(C) Problems to be Solved by the Invention When a glass capillary is used, it is very difficult to capture cells floating in a liquid without damaging them freely. This is because, when the glass capillary is approached, the floating cell escapes by the water flow caused by the glass capillary, and when the negative pressure is increased to adsorb from a far direction, the adsorbing force causes the cell to be broken. This was particularly noticeable for smaller cells.
（ニ） 問題点を解決するための手段 本発明は、液体中に浮遊している細胞を細胞の大きさ
の如何にかかわらず容易にしかも傷つけずに捕獲できる
装置を提供することを目的とする。(D) Means for Solving the Problems The object of the present invention is to provide a device which can easily and without damaging cells floating in a liquid regardless of the size of the cells. .
本発明は、細胞懸濁液を収容するチャンバーと、該チ
ャンバー内に設けられたチャンバー内電極と、チャンバ
ー内で細胞捕獲を行うキャピラリーとからなり、キャピ
ラリーの吸入口と平行な方向の断面の面積がチャンバー
内電極の面積より小さく、高周波電圧の印加によりチャ
ンバー内電極との間で電界効果を生じさせる電極をキャ
ピラリーに内設したことを特徴とする。The present invention comprises a chamber for accommodating a cell suspension, an electrode in the chamber provided in the chamber, and a capillary for capturing cells in the chamber, and has an area of a cross section in a direction parallel to a suction port of the capillary. Is smaller than the area of the electrode in the chamber, and an electrode for generating an electric field effect with the electrode in the chamber by application of a high-frequency voltage is provided inside the capillary.
（ホ） 作用 本発明は、ガラスキャピラリー内の電極と細胞懸濁液
収納チャンバー内の電極との間に高周波電圧が印加され
ると、細胞が電界効果で該キャピラリーの吸入口へ泳動
され捕獲される。(E) Function In the present invention, when a high-frequency voltage is applied between an electrode in a glass capillary and an electrode in a cell suspension storage chamber, cells are migrated and trapped in an inlet of the capillary by an electric field effect. You.
（ヘ） 実施例 本発明の実施例を図面に基づいて説明する。(F) Example An example of the present invention will be described with reference to the drawings.
第１図は本発明に係る細胞捕獲装置を用いて細胞を捕
獲・固定する状態を示す概略図である。FIG. 1 is a schematic diagram showing a state in which cells are captured and fixed using the cell capturing device according to the present invention.
（１）が本発明に係る細胞捕獲装置で、ガラスキャピ
ラリー（７）には電極（６）が埋設されると共にその吸
入口（11）と反対側には負圧発生器たとえば超小型真空
ポンプ（４）が接続されている。このガラスキャピラリ
ー（７）の先すなわち吸入口は細胞の大きさに合わせて
数μｍ〜数10μｍぐらいが適当である。他方、（２）は
細胞懸濁液収納チャンバーで周囲に電極（５）が配設さ
れている。この細胞懸濁液収納チャンバーの材質は、顕
微鏡（10）で細胞懸濁液（９）内の細胞（８）を見るこ
とができるようガラスのような透明体である。(1) A cell capture device according to the present invention, in which an electrode (6) is buried in a glass capillary (7) and a negative pressure generator such as a micro vacuum pump ( 4) is connected. The tip of the glass capillary (7), that is, the inlet is suitably several μm to several tens μm in accordance with the size of the cell. On the other hand, (2) is a cell suspension storage chamber around which an electrode (5) is arranged. The material of the cell suspension storage chamber is a transparent material such as glass so that the cells (8) in the cell suspension (9) can be seen with a microscope (10).
なお電極（６）と電極（５）は高周波電源（３）に接
続されており、また電界強度に差が生じるように電極
（５）は電極（６）より電極断面積が大きい。ここで、
電極（６）の断面の方向は、ガラスキャピラリー（７）
の吸入口（11）と平行の方向であり、この方向の断面積
が電極（５）の縦方向断面積より小さい。電極（５）
（６）の断面積は、電極（５）（６）間で電界強度の勾
配が生じるものならば何でもよく、その大きさはチャン
バーの大きさ等により適宜選択できる。The electrode (6) and the electrode (5) are connected to the high-frequency power supply (3), and the electrode (5) has a larger electrode cross-sectional area than the electrode (6) so as to cause a difference in electric field strength. here,
The direction of the cross section of the electrode (6) is the glass capillary (7).
And the cross-sectional area in this direction is smaller than the vertical cross-sectional area of the electrode (5). Electrode (5)
The cross-sectional area of (6) may be any as long as a gradient of the electric field strength occurs between the electrodes (5) and (6), and the size can be appropriately selected depending on the size of the chamber and the like.
本発明で細胞捕獲を行う条件は、電極（６）を用いる
以外は周知の細胞融合の条件を採用できる。例えば、タ
バコ葉肉プロトプラストを捕獲する場合は市販のシャー
レ（直径8cm、高さ0.2cm）にタバコ葉肉プロトプラスト
を含む懸濁液（2.5mM CaCl2、0.2M ソルビトール、0.
2M マンニトール）を加え、電極（５）（６）間に100V
/cmの高周波電圧を印加する。電極（５）（６）の断面
積は両者間で電界強度に差が生じる大きさならばいくら
でもよいが、シャーレの大きさ、ガラスキャピラリー
（７）の吸入口（11）の径より適宜選択できる。The conditions for performing cell capture in the present invention may employ well-known conditions for cell fusion except for using the electrode (6). For example, when capturing tobacco leaf protoplasts, a suspension (2.5 mM CaCl 2 , 0.2 M sorbitol, 0.2 M sorbitol, containing tobacco leaf protoplasts in a commercially available petri dish (diameter 8 cm, height 0.2 cm)).
2M mannitol) and add 100V between electrodes (5) and (6).
/ cm high frequency voltage is applied. The cross-sectional area of the electrodes (5) and (6) may be any size as long as a difference in electric field strength between the two occurs, but can be appropriately selected from the size of the petri dish and the diameter of the inlet (11) of the glass capillary (7). .
以上の構成において電極（５）（６）に高周波電圧が
印加されると、細胞（８）に分極が生じ電界強度の差に
よって第２図に示すとおりガラスキャピラリー（７）の
吸入口（11）近傍に細胞（８）が泳動する。なお、細胞
の泳動の原理は、誘電電気泳動法による（H.A.Poul:Die
lectrophoresis,Cambridge University Press.,Cambrid
ge（1978）．）。In the above configuration, when a high-frequency voltage is applied to the electrodes (5) and (6), the cells (8) are polarized and the difference in electric field strength causes the inlet (11) of the glass capillary (7) as shown in FIG. The cell (8) migrates in the vicinity. The principle of cell migration is based on dielectrophoresis (HAPoul: Die
electrophoresis, Cambridge University Press., Cambrid
ge (1978). ).
泳動した細胞（８）が吸入口（11）に吸引されると、
負圧発生器（４）を作動させて吸入口（11）に吸着させ
捕獲固定させる。When the electrophoresed cells (8) are sucked into the inlet (11),
The negative pressure generator (4) is actuated to adsorb and capture and fix to the suction port (11).
この状態で他のキャピラリーを用いて細胞（８）内に
DNAを注入したり細胞（８）内から物質を抽出したりす
ることもできる。In this state, use another capillary to enter the cell (8).
It is also possible to inject DNA or extract substances from inside the cells (8).
電極の内設の仕方としてはキャピラリー内面に貼付る
形にしてもよく、また電極をキャピラリー内軸芯方向に
出入れ可能にしてその先端位置を適宜最適位置に調節で
きるような機構を付設してもよい。キャピラリーはガラ
スのみに限定されない。The electrode may be attached to the inner surface of the capillary as a method of installing the electrode, and a mechanism is provided so that the electrode can be inserted and removed in the axial direction of the capillary inside and the tip position can be adjusted to the optimum position as appropriate. Is also good. Capillaries are not limited to glass only.
（ト） 効果 本発明は電界効果で細胞を吸引するため、キャピラリ
ーの動きによる水流の乱れによる細胞の逃避現象は解消
し容易に細胞を捕獲（捕捉）できる。特に小さな細胞の
ときその効果は顕著である。(G) Effect Since the present invention sucks cells by the electric field effect, the escape phenomenon of the cells due to the disturbance of the water flow due to the movement of the capillary is eliminated and the cells can be easily captured (captured). The effect is remarkable especially for small cells.
また、電気の力で軽くキャピラリーの先端に吸着した
後負圧にして固定するもので細胞に大きな力が作用せ
ず、細胞をきずつけず、特に弱い細胞捕獲に有効であ
る。In addition, since it is lightly adsorbed on the tip of the capillary by the force of electricity and then fixed at a negative pressure, a large force does not act on the cells, the cells are not damaged, and it is particularly effective for capturing weak cells.
第１図が本発明に係る細胞捕獲固定装置の概略図、第２
図が細胞が泳動する状態を示す図である。 （１）……細胞捕獲装置、（２）……細胞懸濁液収納チ
ャンバー （３）……高周波電源、（４）……負圧発生器 （５）……チャンバー内電極、（６）……電極 （７）……キャピラリー、（８）……細胞FIG. 1 is a schematic view of a cell capturing and fixing apparatus according to the present invention, and FIG.
The figure shows the state in which cells migrate. (1) ... cell capture device, (2) ... cell suspension storage chamber (3) ... high frequency power supply, (4) ... negative pressure generator (5) ... electrode in chamber, (6) ... … Electrode (7) …… capillary, (8) …… cell
Claims (1)
ャンバー内に設けられたチャンバー内電極と、チャンバ
ー内で細胞捕獲を行うキャピラリーとからなり、キャピ
ラリーの吸入口と平行な方向の断面の面積がチャンバー
内電極の面積より小さく、高周波電圧の印加によりチャ
ンバー内電極との間で電界効果を生じさせる電極をキャ
ピラリーに内設したことを特徴とする細胞捕獲装置。1. A chamber for accommodating a cell suspension, an electrode in the chamber provided in the chamber, and a capillary for capturing cells in the chamber, having a cross section in a direction parallel to a suction port of the capillary. A cell capture device having an area smaller than an area of an electrode in a chamber, and an electrode for generating an electric field effect with the electrode in the chamber when a high-frequency voltage is applied, is provided inside the capillary.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19581886A JP2590459B2 (en) | 1986-08-21 | 1986-08-21 | Cell capture device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19581886A JP2590459B2 (en) | 1986-08-21 | 1986-08-21 | Cell capture device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6352871A JPS6352871A (en) | 1988-03-07 |
| JP2590459B2 true JP2590459B2 (en) | 1997-03-12 |
Family
ID=16347503
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP19581886A Expired - Fee Related JP2590459B2 (en) | 1986-08-21 | 1986-08-21 | Cell capture device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2590459B2 (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3067347B2 (en) * | 1991-10-30 | 2000-07-17 | 株式会社島津製作所 | Gel-like bead sorting equipment |
| US5272926A (en) * | 1991-11-13 | 1993-12-28 | Wilkins Judd R | Microbial retrieval and sampling pipette with a removable cover |
| DE19841337C1 (en) * | 1998-05-27 | 1999-09-23 | Micronas Intermetall Gmbh | Intracellular manipulation of biological cell contents, assisting injection or removal of substances or cell components |
| CN110229752B (en) * | 2019-06-24 | 2024-03-19 | 上海长海医院 | Micro negative pressure cell culture device |
-
1986
- 1986-08-21 JP JP19581886A patent/JP2590459B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6352871A (en) | 1988-03-07 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |