JPH09289886A - Device for detecting cell membrane potential - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable the handling of plural cells at the same time and the quantitative determination of the cell membrane potentials. SOLUTION: A glass microelectrode 6 for thrusting a cell is protruded on the bottom of each well 4 of a part dish 2 and a reference electrode 8 is attached to a side wall of the well. One cell 12 is put into each well 4, the microelectrode 6 is thrust into the cell and the well 4 is filled with a medium 14. The electrodes 6, 8 are connected to an exterior instrument through a connector 10. The membrane potentials of the cells 12 in the individual wells are simultaneously measured based on the medium 14.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an apparatus used for detecting a cell membrane potential in fields such as cytology, biology and biotechnology.

[0002]

2. Description of the Related Art Cell membrane potential is measured as a means for measuring cell activity and the like. Cell membrane potential measurement methods are roughly classified into the following three methods. (1) An intracellular induction method in which a microelectrode is inserted into a cell. In that method, an ion selective electrode and a standard electrode are inserted into one cell, and the potential difference between both electrodes is measured. (2) An intracellular perfusion method (patch clamp method) in which a glass measuring pipette is pressed against a cell membrane to measure ion transport. (3) An optical measurement method which optically measures the degree of fluorescence associated with a membrane potential change using a dye (membrane potential sensitive fluorescent dye) whose fluorescence changes with a membrane potential change.

[0003]

Among the above three measuring methods, the methods (1) and (2) can measure only one cell at a time. Although the method (3) can handle a plurality of cells at the same time, only a qualitative change, that is, a change in fluorescence brightness is measured using a fluorescence microscope, and a quantitative measurement of cell membrane potential is difficult. In addition, all of these methods have a drawback that the measuring device becomes large in size. Therefore, it is an object of the present invention to be able to handle a plurality of cells at the same time, and further to enable quantitative measurement of cell membrane potential.

[0004]

In the cell membrane potential detecting device of the present invention, a dish having a plurality of bottomed holes is provided with a connector for electrical connection to the outside, and each of the holes has a microelectrode protruding to the bottom. And a reference electrode arranged on the side surface are attached, and the microelectrode and the reference electrode in each hole are electrically connected to the connector.

One cell is placed in each hole of the dish and the microelectrode is pierced into the cell. A medium is placed on the cells and the cells and the reference electrode are immersed in the medium. The cell membrane potential of each cell can be measured by connecting to an external device via the connector and measuring the potential difference between the microelectrode and the reference electrode for the cell in each hole.

[0006]

FIG. 1 shows an embodiment, (A) is a plan view,
(B) is a cross-sectional view of one hole, and (X) in (A)
The cross section at the -X 'line position is shown. The petri dish 2 made of transparent plastic is provided with a plurality of holes 4.
The hole 4 has a bottom, and a cell-inserting glass microelectrode 6 is attached to the bottom of each hole so as to project into the hole. A reference electrode 8 is attached to the inner side surface of the hole 4. The petri dish 2 is provided with a connector 10 for electrically connecting to an external device, and the microelectrode 6 and the reference electrode 8 in each hole are electrically connected to the connector 10, and each hole is provided via the connector 10. The electrodes 6 and 8 are electrically connected to an external device.

One cell 12 is placed in each hole 4. The cells 12 are aspirated by, for example, a micropipette and put into the holes 4, and the microelectrodes 6 are inserted into the cells by being pressed against the bottom. After the cells 12 are placed in each hole 4,
Fill all the holes 4 of the dish with medium 14.

FIG. 2 shows cells 12 in each hole 4 of the petri dish 2.
The figure shows a state in which the microelectrode 6 is inserted and the hole 4 is filled with the medium 14, and then the measurement is performed by connecting to an external device. A connector 22 connected to the A / D converter 20 is connected to the connector 10. The A / D converter 20 is connected to the computer 24.

The membrane potential of the cell 12 in each hole 4 of the petri dish 2 is the potential based on the medium 14 as an A / D converter 20.
It is taken into the computer 24 via the, and the membrane potential of the cell 12 in each hole 4 is displayed on the display device of the computer 24.

In this embodiment, since the petri dish 2 is made of transparent plastic and the microelectrodes 6 are glass electrodes, the cell morphology can be observed with a phase contrast microscope at the same time when the cell membrane potential is measured. .. The microelectrode 6 may be a metal electrode when the cells are not observed under a microscope. The reference electrode 8 may be made of any material.
In addition to the cell membrane potential measurement, the measuring device of the present invention can also be used as a device for electrically stimulating the cell 12 by applying a voltage to the cell 12 using the connector 10 as an input.

[0011]

INDUSTRIAL APPLICABILITY In the present invention, the membrane potentials of a plurality of cells can be quantitatively and simultaneously measured. Further, since the electrodes are provided in the holes of the petri dish and the connector is provided in the petri dish, the measuring device can be made compact. In order to measure live cells, the cells must be immersed in the medium,
In the present invention, the medium is filled in each hole of the petri dish, which is convenient for handling live cells.

[Brief description of drawings]

FIG. 1 is a diagram showing an embodiment, (A) is a plan view,
(B) is a sectional view taken along line XX 'of (A).

FIG. 2 is a schematic configuration diagram showing a usage state of the embodiment.

[Explanation of symbols]

 2 Petri dish 4 holes 6 Microelectrode 8 Reference electrode 10 Connector 12 Cells 14 Medium

Claims (1)

[Claims] 1. A petri dish having a plurality of bottomed holes is provided with a connector for making an electrical connection to the outside, and in each of the holes, a microelectrode protruding to the bottom and a reference electrode arranged on a side surface are attached. A cell membrane potential detection device, wherein the microelectrode and the reference electrode of each hole are electrically connected to the connector.