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

外国特許コード F170009193
整理番号 AF19-03WO
掲載日 2017年9月12日
出願国 大韓民国
出願番号 20147016942
公報番号 20140098161
公報番号 101680154
出願日 平成24年12月5日(2012.12.5)
公報発行日 平成26年8月7日(2014.8.7)
公報発行日 平成28年11月28日(2016.11.28)
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
  • 2012JP81556 (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
発明の概要(英語) An object of the present invention is directed to reducing a noise current by suppressing variations in an applied membrane potential in the planar patch clamp device, thereby enabling accurate measurement of an ion channel current. A planar patch clamp device comprising: an electrically insulative substrate having at least one fine through holes; a liquid reservoir and an electrode unit disposed on each of surfaces of the through-hole are configured so that the liquid reservoir retains a conductive liquid and the electrode unit is electrically communicate with the liquid reservoir, wherein the electrode comprises (a) an electrode vessel, wherein at least part of a vessel wall 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 at a saturated concentration, is disclosed.
従来技術、競合技術の概要(英語) BACKGROUND ART
Living entity on the surface of the cells that constitute the various membrane protein to be placed, a chemical substance to the particular sites in the cell surface (such as a ligand of signal transduction material) the binding of the stimulation electrical or light (the gate trigger) membrane protein by radially outward of the channel is opened or closed so between the outside and inside of the cell membrane between a metal ion or a chemical substance in which the transport of the control device and the semiconductor device. Performing this control ion channel is the biological system regarding the delivery signal and an important membrane protein, measurement and its function related to the function of the protein in the channel in the development of drug products that electrical changes, i.e. ion channel current of metrology operations are required.
With respect to this demand, there has heretofore been a patch clamp pipette planar patch clamp techniques such as but has been developed. Pipette patch clamp includes a multipoint measured result in high throughput cannot be applied to screening a weak point of a present invention. With respect to this sample, planar patch clamp includes a silicon chip on a solid substrate such as a plurality of patch clamp apparatus configured and then to be able to make the multipoint measurement, each patch clamp apparatus cell placement sites may include for each ion channel measures the current minute through holes is formed therein.
For example in the following patent document 1 includes a plurality of patch clamp cell patch clamp recording was performed in the following a plurality of electrodes for a planar patch clamp electrode with respect to the arrangement have been described. In addition, the following patent document 2 further includes a silicon substrate to form an electrode and to the surface of the upper limit of, in between the two electrodes in electrical communication with the hole for to penetrate and, to an inlet of the through holes of the two cells for measuring changes in a planar type high voltage electrical patch clamp device is disclosed.
特許請求の範囲(英語) [claim1]
1. (1) the surface of the electrically insulating substrate and the opposite sides thereof are connected to each other, passing through the liquid but not cells capable of fine through holes or less than 1 to form the through holes 1, (2) 2 of the first and the second surface side of each conductive surface side of the liquid the liquid reservoir portion for holding, the conductive liquid and the liquid reservoir electricity to the electrode portion is arranged to form a first liquid reservoir 1, (3) the surface side of the liquid reservoir for the cell arrangement and, wherein the first side and a second surface 2 1 also (4) surface side (a) an electrode portion of the (c) up ∼ element with which said planar patch clamp apparatus. (a) In the liquid reservoir when the conductive liquid is introduced into the conductive liquid and the container wall to abut the at least a portion of the inorganic porous material such as electrode receptacle. (b) the electrode which is contained in the container, (Nm) noble metal on the surface area of the layer is formed of a noble metal (NmCl) chloride electrode. (c) the electrode and is placed in the container, wherein the noble metal chloride (NmCl) and alkali metal salts of saturated solution dissolved with a concentration thereof electrode.

[claim2]
2. Method according to claim 1, wherein the liquid reservoir in the material of the surface side of the light impermeable 1 is constructed by disposing the liquid reservoir portion and a second cell 1, 1 is disposed on the first surface side of the first electrode portion 2 the liquid reservoir portion, the main liquid reservoir and a reservoir communicating these narrow by-pass route mat consisting of a planar patch clamp said device.

[claim3]
3. Method according to claim 1 or 2, wherein the first liquid reservoir 2 the surface side of the conductive liquid path for discharging the introduced and passed through and communicating with, the first electrode portion 2 is also in the liquid pathway is disposed on the surface side of the planar patch clamp apparatus characterized.

[claim4]
4. Method according to claim 1 or 2, wherein the first surface side and the liquid reservoir 1 form a plurality of, each of these nerve cells are arranged in the liquid reservoir to the planar patch clamp apparatus.

[claim5]
5. Method according to claim 4, each of the plurality of concave portions goes down, and the concave portion and a concave portion and the connecting groove, wherein the recess is disposed in a nerve cell, its movement can be blocked with a depth and size, the formation of the grooves of the nerve cells to the network of the planar patch clamp apparatus.

[claim6]
6. Method according to claim 1 or 2, wherein the planar patch clamp apparatus and/or the following (A) (B) is provided with the configuration of the planar patch clamp apparatus. (A) the surface side of the first liquid suction device 2 connected to the liquid reservoir and, the suction device 2 by the liquid in the liquid reservoir of the negative pressure surface side of the load to be un. (B) Through hole 1 of the substrate in the second surface side of the rim of the opening of the holding force on the cells having adhered to the extracellular matrix forming material present.

[claim7]
7. AMEND STATUS: Delete

[claim8]
8. AMEND STATUS: delete

[claim9]
9. AMEND STATUS: delete

[claim10]
10. AMEND STATUS: delete

[claim11]
11. The first 2 or 1 based on the first surface of the planar patch clamp apparatus 2 on the side of the first introducing a liquid in the liquid reservoir 1 and the conductive surface side of the cells to be measured goes dispersed in the conductive liquid is introduced into the liquid reservoir portion 1 of the first surface side of the first passage to the liquid reservoir 2 and the surface side of the conductive, wherein the cells with a second liquid reservoir 1 arranged in a position of a predetermined surface side in a state of the surface side and a second surface side of the 1 2 by applying a voltage between the electrodes of the electrode portion to be measured and measures the current cell of the ion channel of the ion channel current cell measurement method.

[claim12]
12. Method according to claim 11, wherein the planar patch clamp apparatus 1 includes a plurality of first surface side of the substrate of the liquid reservoir is formed, to be measured is a neural cell and the cell, the cell culture conductive liquid is a neuronal cell ion channel current measurement method.

[claim13]
13. Method according to claim 12, wherein said neural cells include for imaging is introduced Ca Ca in advance and the probe, at least an action potential in the cell current ion channel activity upon the occurrence of the potential of the cell or generated during the propagation of the fluorescence imaging means including Ca observed by said cell measuring method for measuring ion channel current.

[claim14]
14. Method according to claim 13, wherein the probe is introduced into the Ca planar patch clamp apparatus nerve cells of the first plurality 1 to be arranged on a surface side thereof a plurality of nerve cells are also concatenated network synaptic nerve cells and the piece of, single one of these current injection or voltage nerve cells by applying a plurality of nerve cells to a plurality of said Ca by imaging the measurement was carried out in a cell current measurement method of the ion channel.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • URISU TSUNEO
  • WANG ZHI HONG
  • UNO HIDETAKA
  • OBULIRAJ SENTHIL KUMAR
  • NAGAOKA YASUTAKA
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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