TOP > 外国特許検索 > Planar patch clamp device, electrodes for said device and cell ion channel current measurement method

Planar patch clamp device, electrodes for said device and cell ion channel current measurement method NEW

外国特許コード F170009186
整理番号 AF19-03WO
掲載日 2017年9月12日
出願国 アメリカ合衆国
出願番号 201214366708
公報番号 20140339102
出願日 平成24年12月5日(2012.12.5)
公報発行日 平成26年11月20日(2014.11.20)
国際出願番号 JP2012081556
国際公開番号 WO2013094418
国際出願日 平成24年12月5日(2012.12.5)
国際公開日 平成25年6月27日(2013.6.27)
優先権データ
  • 特願2011-278445 (2011.12.20) JP
  • 特願2012-181786 (2012.8.20) JP
  • 2012WO-JP81556 (2012.12.5) WO
発明の名称 (英語) Planar patch clamp device, electrodes for said device and cell ion channel current measurement method NEW
発明の概要(英語) (US20140339102)
An object of the present invention is that variations in an applied membrane potential in the planar patch clamp device are suppressed to reduce a noise current, thereby enabling accurate measurement of an ion channel current.
Disclosed is a planar patch clamp device including: an electrically insulative substrate provided with one or more fine through holes; a liquid reservoir that holds a conductive liquid provided on both surface sides of the through hole; and energizable electrode sections provided in the liquid reservoir; these electrode sections including: (a) an electrode vessel, at least part of which is made of an inorganic porous material, (b) an electrode in which a chloride NmCl layer is formed on the surface of a noble metal Nm, and (c) an electrode solution containing NmCl and an alkali metal chloride being dissolved therein at a saturated concentration.
特許請求の範囲(英語) [claim1]
1. A planar patch clamp device comprising: (1) an electrically insulative substrate provided with one or more fine through holes, each having an inner diameter through which a cell cannot pass but a liquid can pass, the fine through holes communicating between surfaces on both sides of the electrically insulative substrate; (2) a liquid reservoir that holds a conductive liquid, and an electrode section disposed energizably for a conductive liquid in the liquid reservoir, provided on a first surface side and a second surface side of the through hole; (3) the liquid reservoir on the first surface side serving as a liquid reservoir for cell placement; and (4) the following constituents (a) to (c) included in electrode sections on the first surface side and second surface side: (a) an electrode vessel, at least part of which is made of an inorganic porous material that comes into contact with a conductive liquid when introduced into the liquid reservoir,
(b) an electrode accommodated in the electrode vessel, noble metal chloride NmCl layer being formed on the surface of the noble metal Nm, and
(c) an electrode solution filled in the electrode vessel, the noble metal chloride NmCl and an alkali metal chloride being dissolved therein at a saturated concentration.
[claim2]
2. The planar patch clamp device according to claim 1, wherein the liquid reservoir on the first surface side includes a primary liquid reservoir for cell placement, made of a light non-transmissive material, a secondary liquid reservoir provided with an electrode section on the first surface side, and a narrow liquid path that communicates between the primary liquid reservoir and the secondary liquid reservoir.
[claim3]
3. The planar patch clamp device according to claim 1 or 2, wherein the liquid reservoir on the second surface side is communicated with a liquid path to introduce and discharge a conductive liquid, and an electrode section on the second surface side is disposed on this liquid path.
[claim4]
4. The planar patch clamp device according to any one of claims 1 to 3, wherein a plurality of liquid reservoirs are disposed on the first surface side, and a recess having a width larger than that of a cell body of a nerve cell and having a depth that can inhibit the movement of a cultured cell body is formed in these liquid reservoirs.
[claim5]
5. The planar patch clamp device according to any one of claims 1 to 4, including the following constitution(s) (A) and/or (B): (A) a liquid reservoir on the second surface side is communicated with a liquid suction device, thereby enabling negative pressure load on the liquid reservoir on the second surface side, and
(B) an extracellular matrix-forming substance having cell fixing capability is attached on the circumference of the opening on the first surface side of a through hole on a substrate.
[claim6]
6. An electrode section for planar patch clamp device, comprising the following constituents (a') to (c'): (a') an electrode vessel, wherein at least part of a vessel wall is made of an inorganic porous material,
(b') an electrode accommodated in the electrode vessel, noble metal chloride NmCl layer being formed on the surface of the noble metal Nm, and
(c') an electrode solution filled in the electrode vessel, the noble metal chloride NmCl and an alkali metal chloride being dissolved therein at a saturated concentration.
[claim7]
7. The electrode section for planar patch clamp device according to claim 6, wherein the inorganic porous material composing at least part of the vessel wall is porous glass or porous ceramic.
[claim8]
8. The electrode section for planar patch clamp device according to claim 6 or 7, wherein a noble metal Nm is silver Ag or platinum Pt, and an alkali metal chloride is potassium chloride KCl.
[claim9]
9. The electrode section for planar patch clamp device according to any one of claims 6 to 8, wherein an electrode is the following (c) or (D): (C) a stick electrode protruding inside an electrode vessel, a noble metal chloride NmCl layer being formed on the surface of a core made of the noble metal Nm, or
(D) a cylindrical electrode formed on the inner periphery of an electrode vessel wall, wherein the bottom layer on the vessel wall side is a deposited layer of a noble metal Nm and the surface layer in contact with an electrode solution is a deposited layer of a noble metal chloride NmCl.
[claim10]
10. A cell ion channel current measurement method, which comprises introducing a conductive liquid into a liquid reservoir on a second surface side of the planar patch clamp device according to any one of claims 1 to 5, and introducing a conductive liquid containing a cell to be measured dispersed therein into a liquid reservoir on the first surface side to conductively communicate between the liquid reservoir on first surface side and the liquid reservoir on the second surface side; disposing the cell at a prescribed position of the liquid reservoir on the first surface side; applying a voltage between electrodes of electrode sections on the first surface side and second surface side; and measuring an ion channel current of a cell to be measured.
[claim11]
11. The cell ion channel current measurement method according to claim 10, wherein a plurality of liquid reservoirs of the constitution mentioned in claim 4 are disposed on the first surface side of a substrate in the planar patch clamp device and a cell to be measured is a nerve cell, and a conductive liquid is a cell culture medium of the nerve cell.
[claim12]
12. The cell ion channel current measurement method according to claim 11, wherein a Ca probe for Ca imaging is introduced into a nerve cell in advance, and an ion channel current of the cell is measured by means at least including Ca imaging, which is observation of fluorescence that occurs at generation of cell action potential or at propagation of an action potential.
[claim13]
13. The cell ion channel current measurement method according to claim 12, wherein plural or many nerve cells into which a Ca probe introduced are disposed on the first surface side in the planar patch clamp device, and synapses of these nerve cells mutually linked are defined as a nerve cell network, and a current is injected or a voltage is applied to a single nerve cell of these cells to perform measurement by the Ca imaging in plural or many nerve cells.
  • 発明者/出願人(英語)
  • URISU TSUNEO
  • WANG ZHI-HONG
  • UNO HIDETAKA
  • OBULIRAJ SENTHIL KUMAR
  • NAGAOKA YASUTAKA
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
参考情報 (研究プロジェクト等) CREST Creation of Nanosystems with Novel Functions through Process Integration AREA
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