Planar Patch Clamp Device, Electrodes For Said Device And Cell Ion Channel Current Measurement Method
|発明の名称 （英語）||Planar Patch Clamp Device, Electrodes For Said Device And Cell Ion Channel Current Measurement Method|
In order to reduce a noise current by suppressing variations in an applied membrane potential in a planar patch clamp device, enabling an ion channel current to be accurately measured, this planar patch clamp device is provided with a liquid reservoir that has at least one fine through-hole disposed on an electrically insulative substrate, and holds a conductive liquid on surfaces on both sides of the through-hole; and electrodes, which are capable of being energized, are disposed in said liquid reservoir.
The electrodes are provided with: (a) an electrode vessel, wherein part of the vessel wall comprises an inorganic porous material; (b) an electrode, wherein a noble metal chloride (NmCl) layer is formed on the surface of the noble metal (Nm); and (c) an electrode solution, wherein the noble metal chloride and an alkali metal chloride are dissolved at a saturated concentration.
1. A planar patch clamp device, characterized in,
1 electrically insulating(1) substrates more fine through holes, the through hole communication with the lateral surface of the electrically insulating substrate while not capable by the cells passing through the liquid, (2) with the through hole of the side surface 2nd 1st are provided: a conductive liquid for maintaining ; and configured to be a conductive liquid on the electrode part, (3) the 1st surface is configured as the cells , the surface 2nd of the 1st surface side and further(4) comprising a side electrode (a)-(c) elements:
(A) with the electrode reservoir, with the conductive liquid to the conductive liquid into contact at least a portion of the vessel wall made of an inorganic porous material;
(B) electrode, which is contained in the electrode reservoir within the surface layer portion formed of noble metal Nm noble metal chloride NmCl layer; and
(C) electrode solution, which is filled in the electrode holder and to the saturated concentration of dissolved chloride NmCl noble metal and alkali metal chloride.
2. Planar patch clamp device according to claim 1, characterized in,
1st the surface of the sump 1st, 2nd and the reservoir and to pass these defining narrow communication, from the reservoir 1st for configuring cells and the light non-transmitting material, the electrode portion 1st disposed side reservoir 2nd.
3. Planar patch clamp device according to claim 1 or 2, characterized in,
2nd the surface side of the conductive liquid and for introducing and exhausting of flow passage in communication with, and in the flow channel 2nd disposed on the surface side electrode portion.
4. 1 according to claim-planarly a patch clamp apparatus any one of claims 3, characterized in,
1st in a plurality of surface side , in these neural cells are formed is larger than the width and the depth of the cell culture state to prevent movement of the recess.
5. 1 according to claim-plane in a patch clamp apparatus according to any of claims 4, characterized in,
A planar patch clamp device includes (A) and a/or (B) consisting of:
(A) 2nd surface side of the liquid suction means is connected, through the liquid suction device capable of applying a negative pressure side 2nd;
(B) has the fixing force of the extracellular matrix-forming material attached to the substrate through hole 1st of the side opening perimeter.
6. A planar patch electrode means, characterized in,
Further comprising a step of (a ')-(c') of the element:
(A ') of the vessel wall is composed of inorganic porous material of at least one electrode reservoir;
(B') is housed in the electrode holder and a surface layer portion formed in the noble metal Nm NmCl layer of noble metal chloride electrode; and
(C') dissolving a saturating concentration for the electrode reservoir is filled inside with an alkali metal chloride noble metal chloride NmCl electrode solution.
7. Planar patch electrode device according to claim 6, characterized in,
At least a portion of the container wall is formed of an inorganic porous material is a porous glass or ceramic.
8. Planar patch clamp device electrode 6 according to claim or 7, characterized in,
A noble metal or platinum Pt Ag Nm of silver,
Alkali metal chloride is potassium chloride KCl.
9. 6 according to claim-plane clamp device electrode any one of claims 8, characterized in,
A electrode (C) or (D):
(C) a stick electrode is the electrode inside the container, a noble metal Nm of the core layer is formed of noble metal chloride NmCl surface,
(D) is formed in the wall of the cylindrical inner peripheral surface of the electrode reservoir electrode, the container wall portion side of the layer is a noble metal deposited Nm, in contact with the electrode solution of the deposition layer is a noble metal chloride NmCl.
10. A cell ion channel current measurement method, characterized in,
Claim 1-5 to, the planar patch clamp device according to any one of the conductive liquid into 2nd surface side, and the introduction of the 1st surface to be measured dispersed conductive liquid, the surface side of 1st to 2nd of the surface side of the conductive communication, the cells arranged in a predetermined position of the 1st surface , on the basis of the surface side and side surface of the electrode 1st 2nd applying a voltage between the electrodes, whereby the cell current as a measurement object measuring channel.
11. Cell ion channel current measurement method according to claim 10, characterized in,
1st the substrate of the planar patch clamp device having a surface provided with a plurality of according to claim 4,
The cells are nerve cells to be measured,
The conductive liquid is a neural cell culture.
12. Cell ion channel current measurement method according to claim 11, characterized in,
A lead to the nerve cells Ca Ca probe for imaging,
Image forming means including at least measured Ca of ion channel current cells, which cells Ca action potential propagation time of imaging is observed for the occurrence of the fluorescence generated action potentials.
13. Cell ion channel current measurement method according to claim 12, characterized in,
1st the clamp device is disposed in the planar surface into which a plurality of the probe even more Ca neural cells, neural cells and form the neural network synapses are connected, to the nerve cells of a cell of a single current injection or voltage application, thereby to more even more neural cells in the measurement based on the imaged Ca.
|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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