JP2020080249A - Circular electron emission element and cell stimulation device - Google Patents

Abstract

To provide a circular electron emission element capable of effectively securing an electron emission region of the electron emission element and improving uniformity of an electron emission amount in the electron emission region.SOLUTION: A circular electron emission element comprises: a lower electrode; a surface electrode facing the lower electrode; an intermediate layer disposed between the lower electrode and the surface electrode; and an insulation layer including an opening. The insulation layer is disposed between the lower electrode and the intermediate layer or between the intermediate layer and the surface electrode. The lower electrode, the intermediate layer and the surface electrode are provided in such a manner that a potential difference is generated between the lower electrode and the surface electrode, thereby causing a current to flow to the intermediate layer and emitting electrons from an electron emission region of the surface electrode. The electron emission region is defined by the opening of the insulation layer, and the opening is a circle or a partially missed circle.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a circular electron emitting device and a cell stimulating device.

A cell culture method is known in which electric stimulation is applied to cultured cells that respond to electric stimulation (see, for example, Patent Document 1). In this cell culture method, cell stimulation is promoted by electrical stimulation, and the cell growth rate can be increased.
On the other hand, a rectangular electron-emitting device is known in which electrons are caused to flow into the intermediate layer by applying a voltage between the lower electrode and the surface electrode to emit the electrons (see, for example, Patent Document 2).

JP-A-60-110287 JP, 2016-136485, A

In the conventional rectangular electron-emitting device, the amount of electron emission at the corner of the rectangular electron-emitting region may be lower than that at the center of the electron-emitting region, or the necessary electron-emitting region may not be sufficiently secured.
The present invention has been made in view of such circumstances, and it is possible to improve the uniformity of the amount of electron emission in the electron emission region, and by ensuring the electron emission region to the maximum, the total electron emission amount of the device is obtained. A circular electron-emitting device capable of increasing

  The present invention comprises a lower electrode, a surface electrode facing the lower electrode, an intermediate layer arranged between the lower electrode and the surface electrode, and an insulating layer having an opening, wherein the insulating layer is It is arranged between the lower electrode and the intermediate layer or between the intermediate layer and the surface electrode, and the lower electrode, the intermediate layer and the surface electrode are between the lower electrode and the surface electrode. A current flows through the intermediate layer by generating a potential difference, and the electron emission region of the surface electrode is provided so that electrons are emitted. The electron emission region is defined by the opening of the insulating layer, and the opening is formed. Provides a circular electron-emitting device having a circular shape or a partially lacking circular shape.

  In the circular electron emitting device of the present invention, the opening of the insulating layer that defines the area of the electron emitting region is circular or partially circular. Therefore, the electron emission region does not have a corner where the amount of electron emission decreases. Therefore, the uniformity of the amount of electron emission in the electron emission region can be improved.

It is a schematic plan view of a circular electron-emitting device according to an embodiment of the present invention. It is a schematic sectional drawing of the circular electron emission element in dashed line AA of FIG. (A)-(c) is a schematic plan view of the circular electron emission element of one embodiment of the present invention. It is a schematic sectional drawing of the circular electron emission element of one embodiment of the present invention. It is a schematic perspective view of the cell stimulator of one embodiment of the present invention. (A)-(d) is a schematic plan view of the cell stimulator of one Embodiment of this invention, respectively. It is a schematic sectional drawing of the cell stimulator of one embodiment of the present invention. It is a schematic plan view of the cell stimulator of one embodiment of the present invention. It is a schematic perspective view of the culture container contained in the cell stimulator of one embodiment of the present invention. It is a schematic sectional drawing of the cell stimulator of one embodiment of the present invention.

The circular electron emission device of the present invention includes a lower electrode, a surface electrode facing the lower electrode, an intermediate layer arranged between the lower electrode and the surface electrode, and an insulating layer having an opening, The insulating layer is disposed between the lower electrode and the intermediate layer or between the intermediate layer and the surface electrode, and the lower electrode, the intermediate layer and the surface electrode are the lower electrode and the surface. A current flows through the intermediate layer by generating a potential difference between the electrode and the electrode, and the electron emission region of the surface electrode is provided so that electrons are emitted, and the electron emission region is formed by the opening of the insulating layer. The opening is defined as a circle or a partially cut circle.
The lower electrode, the intermediate layer, and the surface electrode included in the circular electron-emitting device of the present invention are preferably circular, partially lacking circular or ring-shaped, and the insulating layer has a ring-shaped shape surrounding the opening. It is preferable to have. As a result, the circular electron-emitting device can have a circular shape, a partially lacking circular shape, or an annular shape.

The present invention also provides a cell stimulating device including the circular electron-emitting device of the present invention and a charge recovery electrode. The circular electron-emitting device has a circular shape, a partially lacking circular shape or an annular shape, and is provided so as to supply an electric charge to a medium for culturing cells via a gas phase, and the charge recovery electrode is , To recover the charge from the medium.
With this cell stimulator, it is possible to supply an electric charge to a wide range of the medium in the circular well, and to electrically stimulate many cells cultured in the medium. Further, since the uniformity of the electron emission amount of the circular electron-emitting device can be improved, it is possible to give highly homogeneous electric stimulation to the cells cultured in the medium.

The cell stimulator of the present invention preferably comprises a cylindrical base material, and the circular electron-emitting device is preferably removably fixed on the upper surface of the base material. As a result, the electron emission region of the circular electron emission device can face the surface of the medium in the circular well in a wide range via the gas phase. This allows electrical stimulation to be applied to a wide range of cells in the medium.
It is preferable that the circular electron-emitting device has a circular shape with a part cut off, the charge recovery electrode is a pin electrode or a rod electrode, and protrudes from the base material in a region where the circular electron-emitting device is cut off. This makes it possible to increase the size of the circular electron emitting device arranged on the upper surface of the base material and increase the size of the electron emitting region. As a result, electrons can be emitted toward a wider area of the medium, and more cells can be electrically stimulated.

It is preferable that the circular electron-emitting device has an annular shape, the charge recovery electrode is a pin electrode or a rod electrode, and the charge-collecting electrode projects from the base material at an opening at the center of the circular electron-emitting device. This makes it possible to increase the size of the circular electron emitting device arranged on the upper surface of the base material and increase the size of the electron emitting region. As a result, electrons can be emitted toward a wider area of the medium, and more cells can be electrically stimulated. Further, the charge recovery electrode can be arranged in the center of the medium, and the homogeneity of the electrical stimulation given to the cells can be improved.
The charge recovery electrode preferably has a cylindrical shape or a partially lacking cylindrical shape, and preferably projects from the base material so as to surround the circular electron-emitting device. As a result, the charge recovery electrode can be arranged along the inner wall surface of the circular well, and the homogeneity of the electrical stimulation applied to the cells can be improved.

The charge recovery electrode is preferably an electrode partially covered with an insulator layer, and the insulator layer is preferably disposed between the circular electron emission device and the charge recovery electrode. As a result, it is possible to suppress the occurrence of leak current between the charge recovery electrode and the circular electron-emitting device.
The cell stimulation device of the present invention preferably includes a power supply device. It is preferable that the power supply device is provided so that a potential difference can be generated between the lower electrode and the surface electrode, and a potential difference can be generated between the circular electron-emitting device and the charge recovery electrode. Is preferably provided in the. As a result, an electric field can be generated in the intermediate layer between the lower electrode and the surface electrode, and electrons can be emitted from the electron emission region of the circular electron emission device. Further, the electrons can charge the gas phase component to generate anions. In addition, an electric field can be generated in the gas phase between the medium connected to the charge recovery electrode and the surface electrode, and the anions generated in the gas phase can be supplied to the medium. Anions in the medium generated from the ions can provide electrical stimulation to the cells in the medium.
The cell stimulator of the present invention preferably comprises a culture vessel having a circular well for containing a medium, and the circular electron-emitting device has a surface electrode sandwiching a vapor phase with the surface of the medium contained in the well. It is preferable that they are arranged to face each other.

  Hereinafter, the present invention will be described in more detail with reference to a plurality of embodiments. The configurations shown in the drawings and the following description are examples, and the scope of the present invention is not limited to those shown in the drawings and the following description.

First Embodiment FIG. 1 is a schematic plan view of a circular electron-emitting device of this embodiment, and FIG. 2 is a schematic cross-sectional view of the circular electron-emitting device taken along the broken line AA in FIG. In addition, FIGS. 3A to 3C are schematic plan views of modified examples of the circular emitting device according to the present embodiment.
The circular electron emission device 20 of this embodiment has a lower electrode 2, a surface electrode 3 facing the lower electrode 2, an intermediate layer 4 arranged between the lower electrode 2 and the surface electrode 3, and an opening 6. The insulating layer 5 is disposed between the lower electrode 2 and the intermediate layer 4, and the lower electrode 2, the intermediate layer 4 and the surface electrode 3 are disposed between the lower electrode 2 and the surface electrode 3. A current is caused to flow in the intermediate layer 4 by generating a potential difference, and electrons are provided to be emitted from the electron emission region 7 of the surface electrode 3. The electron emission region 7 is defined by the opening 6 of the insulating layer 5. No. 6 is characterized by being a circular shape or a partially lacking circular shape.

The circular electron emission element 20 is an element that emits electrons from the electron emission region 7 into the air, the gas, the reduced pressure atmosphere, or the like. The electrons emitted from the electron emission region 7 generate anions that charge the gas to the cathode in the air, the gas, the reduced pressure atmosphere, or the like. The circular electron emission element 20 is a cell stimulator, a charging device that charges an object to be charged, an electron beam curing device that cures the object to be cured, a blower, a cooling device, and a self-luminous device that emits light from a light-emitting body. It can be used for an image display device provided with the light emitting device.
The shape of the circular electron-emitting device 20 may be a circular shape having a high circularity as shown in FIG. 1 or a partially lacking circular shape as shown in FIG. As shown in FIG. 3(b), it may have an annular shape having an opening 16 at the center, may have an elliptical shape as shown in FIG. 3(c), and may have a distorted circular shape. Good. Further, the surface electrode 3, the lower electrode 2 and the intermediate layer 4 may also have a circular shape, a partially lacking circular shape, an annular shape or an elliptical shape, depending on the shape of the circular electron emitting device 20. As a result, the electron emission region 7 can be formed in a wide area on the upper surface of the circular electron emission device 20.
The shape of the circular electron-emitting device 20 may be a flat plate shape having a thickness of 200 μm or more and 2 mm or less.

The surface electrode 3 is an electrode located on the surface of the circular electron-emitting device 20. The surface electrode 3 can have a thickness of 5 nm or more and 100 nm or less, preferably 40 nm or more and 100 nm or less. The material of the surface electrode 3 is, for example, gold or platinum. Further, the surface electrode 3 may be composed of a plurality of metal layers.
Even when the surface electrode 3 has a thickness of 40 nm or more, it may have a plurality of openings, gaps, and a portion thinned to a thickness of 10 nm or less. The electrons flowing through the intermediate layer 4 can pass or pass through the openings, the gaps, and the thinned portions, and the electrons can be emitted from the electron emission region 7 of the surface electrode 3. Such openings, gaps, and thinned portions can be formed by applying a voltage between the lower electrode 2 and the surface electrode 3 (forming process, initial voltage application).

  The lower electrode 2 is an electrode that faces the surface electrode 3 with the intermediate layer 4 interposed therebetween. The lower electrode 2 may be a metal plate, or a metal layer or a conductor layer formed on an insulating substrate. When the lower electrode 2 is made of a metal plate, this metal plate may be the substrate of the circular electron-emitting device 20. The material of the lower electrode 2 is, for example, aluminum, stainless steel, nickel or the like. The thickness of the lower electrode is, for example, 200 μm or more and 2 mm or less.

  The intermediate layer 4 is a layer in which a current flows due to an electric field formed by the potential difference between the surface electrode 3 and the lower electrode 2. The intermediate layer 4 can have semiconductivity. The intermediate layer 4 can include at least one of an insulating resin, a conductive resin, and insulating fine particles. Further, the intermediate layer 4 preferably contains conductive fine particles. The thickness of the intermediate layer 4 can be, for example, 1 μm or more and 3 μm or less. Since the electrons that have flowed through the intermediate layer 4 are emitted from the electron emission region 7 of the surface electrode 3 to the gas phase, the circular electron emission device 20 can surface-emit the electrons from the electron emission region 7. Therefore, electrons can be uniformly emitted to the gas phase on the electron emission region 7 of the surface electrode 3. This electron can charge the gas phase component, and can generate an ionic species such as an anion of oxygen in the gas phase.

An insulating layer 5 having an opening 6 is arranged between the lower electrode 2 and the intermediate layer 4. The insulating layer 5 is a layer for defining the electron emission region 7. By defining the electron emission region 7 using the insulating layer 5, it is possible to suppress the concentration of current locally flowing in the intermediate layer 4.
In the region where the insulating layer 5 is arranged between the lower electrode 2 and the surface electrode 3, no current flows in the insulating layer 5, so even if a potential difference is generated between the lower electrode 2 and the surface electrode 3, the intermediate layer No current flows through 4. Therefore, electrons are not emitted in this region, and the region does not become the electron emission region 7. On the other hand, in the region where the opening 6 is arranged, the insulating layer 5 is not arranged between the lower electrode 2 and the surface electrode 3. Therefore, when a potential difference is generated between the lower electrode 2 and the surface electrode 3, the intermediate layer 4 is formed. A current flows through the surface electrode 3, and electrons are emitted from the surface electrode 3 to the gas phase. Therefore, this area becomes the electron emission area 7.
As described above, the electron emission region 7 can be defined by the opening 6 provided in the insulating layer 5, and the position of the opening 6 and the electron emission region 7 substantially coincide with each other.

The opening 6 of the insulating layer 5 has a circular shape, a partially lacking circular shape, an elliptical shape, a distorted circular shape, or the like. Further, the insulating layer 5 may have an annular shape that surrounds the opening 6. The diameter of the opening 6 can be, for example, 0.70 times or more and 0.95 times or less the diameter of the insulating layer 5.
For example, the opening 6 can have a shape as shown by a dotted line in FIGS. 1 and 3A to 3C. As a result, the electron emitting region 7 can be formed into a circular shape, a partially lacking circular shape, an elliptical shape, a distorted circular shape, or the like, and a corner portion having an angle of 90° or less is not formed in the electron emitting area 7.
When the electron emission region 7 has an angle of 90° or less, the corners of the electron emission region 7 are located close to the two sides forming the corner, the influence of the insulating layer 5 becomes large, and the amount of electron emission is different. It is considered to be less than the part. Therefore, it is considered that non-uniformity of the amount of electron emission occurs between the corners of the electron emission region 7 and other regions.
In the present embodiment, since the corner portion having an angle of 90° or less is not formed in the electron emission region 7, it is possible to suppress the generation of a region in which the electron emission amount is reduced, and the electron emission amount in the electron emission region 7 is reduced. Uniformity can be improved.

The insulating layer 5 may be electrically insulating, and the material of the insulating layer 5 is, for example, an inorganic material such as a metal oxide or a metal nitride, or an organic material such as a silicone resin or a phenol resin. Can be used.
The contact point between the surface electrode 3 and the surface electrode connection terminal can be arranged on the insulating layer 5. As a result, it is possible to prevent the current from flowing in the intermediate layer 4 immediately below the contact point, and to prevent the surface electrode 3 from breaking at the contact point.

Second Embodiment FIG. 4 is a schematic cross-sectional view of the circular electron emission device 20 of this embodiment.
The circular electron emission device 20 of the present embodiment includes a lower electrode 2, a surface electrode 3 facing the lower electrode 2, an intermediate layer arranged between the lower electrode 2 and the surface electrode 3, and an insulation having an opening 6. The insulating layer 5 is disposed between the intermediate layer 4 and the surface electrode 3, and the lower electrode 2, the intermediate layer 4 and the surface electrode 3 have a potential difference between the lower electrode 2 and the surface electrode 3. Current is caused to flow in the intermediate layer 4 to cause electrons to be emitted from the electron emission region 7 of the surface electrode 3, and the opening 6 has a circular shape or a partially lacking circular shape. It is characterized in that the range of the emission area 7 is determined.
In the second embodiment, the insulating layer 5 having the opening 6 is arranged between the intermediate layer 4 and the surface electrode 3. The opening 6 of the insulating layer 5 can define the electron emission region 7 of the circular electron emission device 20.
Other configurations are similar to those of the first embodiment. The description of the first embodiment applies to the second embodiment unless there is a contradiction.

Third Embodiment The third embodiment relates to a cell stimulator.
FIG. 5 is a schematic perspective view of the cell stimulating device of the present embodiment, and FIGS. 6A to 6D are schematic top views of the cell stimulating device of the modified example. FIG. 7 is a schematic cross-sectional view of a cell stimulator in a state of electrically stimulating cells cultured in a medium.
The cell stimulator 30 of the present embodiment includes a circular electron emission element 20 and a charge recovery electrode 8, and the circular electron emission element 20 has a circular shape, a partially lacking circular shape or an annular shape, and a cell. It is characterized in that the medium 10 for culturing 11 is provided so as to supply an electric charge via the gas phase 15, and the charge recovery electrode 8 is provided so as to recover the electric charge from the medium 10.
The cell stimulator 30 of the present embodiment includes a base material 13, a surface electrode connecting terminal 25, lower electrode connecting terminals 26a and 26b (these are collectively referred to as lower electrode connecting terminal 26), a lid member 27, an insulating layer 14, The power supply device 21a, 21b (these are collectively referred to as the power supply device 21) or the culture vessel 23 can be provided. The power supply device 21 may be one power supply device or may be composed of a plurality of power supply devices.

The cell stimulating device 30 is a device for applying electrical stimulation to the cells 11 cultured in the medium 10 for the purpose of inducing proliferation and differentiation of the cells 11.
The cells 11 cultured in the medium 10 are, for example, cells/tissues of multicellular organisms, cells into which a gene has been introduced, cells into which a gene has been recombined, cancer cells, microorganisms, and the like. The cells 11 can be cells that respond to electrical stimulation. Examples of cells that respond to electrical stimulation include muscle cells, nerve cells, osteoblasts, osteoclasts, chondrocytes, osteocytes, fibroblasts and the like.
Further, as the cells 11 to be cultured in the medium 10, cells preliminarily precultured can be used. The pre-cultured cells can be seeded in the medium 10 so that the cells 10 are contained in the medium 10 at 10 ³ to 10 ⁶ cells/cm ² .
The medium 10 provides a growth environment for the cells 11. The medium 10 can be used without particular limitation as long as it is a commonly used cell culture medium. The medium 10 may be a liquid medium or a solid medium obtained by solidifying the liquid medium with a gelling agent such as agar. Further, the medium 10 may be a combination of a liquid medium and a solid medium. The medium 10 is, for example, MEM medium (Eagle minimum essential medium), D-MEM medium (Dulbecco modified Eagle medium), α-MEM medium (Eagle minimum essential medium α modified type), GMEM (Glasgow minimum essential medium), Ham's F- 12 (nutrient mixture F-12 ham), IMDM (Iscove's modified Dulbecco's medium), RPMI-1640 medium, D-PBS (Dulbecco phosphate buffered saline), HBSS (Hank's balanced salt solution) and the like.

The medium 10 is contained in the culture container 23. The culture container 23 is, for example, a multi-well plate including a plurality of wells 24, a culture dish, or the like. The well 24 and the culture dish of the multi-well plate usually have a circular shape. Further, a cell culture device can be formed by combining the culture container 23 and the cell stimulating device 30. As the material of the culture container 23, existing materials such as glass and plastic can be used. It is preferable that the culture container 23 has a container bottom surface-treated so as to be suitable for fixing the cells 11. The surface treatment may be a generally known treatment for applying a matrix protein, collagen, fibronectin, laminin or the like to the bottom surface of the container.
The cell culture device can be placed in an incubator. As a result, the temperature of the medium 10, atmospheric gas, etc. can be controlled, and the cells 11 can be stably cultured. The culture atmosphere is, for example, temperature: 37° C., relative humidity: 95%, atmosphere gas: air containing 5% carbon dioxide gas.

  The cell stimulator 30 includes a circular electron emitting element 20. The circular electron emitting device 20 is the circular electron emitting device 20 of the first or second embodiment. The description of the first or second embodiment applies to the circular electron emission device 20 of the third embodiment. The circular electron emitting device 20 is a device having a planar shape and capable of emitting electrons from the electron emitting region 7. The circular electron emitting device 20 includes a lower electrode 2, a surface electrode 3, and an intermediate layer 4 arranged between the lower electrode 2 and the surface electrode 3. The electron emission region 7 is defined by the opening 6 of the insulating layer 5 arranged between the lower electrode 2 and the intermediate layer 4 or between the intermediate layer 4 and the surface electrode 3.

The circular electron-emitting device 20 is provided so as to supply an electric charge to the medium 10 for culturing the cells 11 via the gas phase 15. Further, the circular electron-emitting device 20 can be arranged so that the surface electrode 3 faces the surface of the medium 10 housed in the well 24 of the culture container 23 with the gas phase 15 interposed therebetween. The distance between the surface electrode 3 and the surface of the medium 10 can be 0.5 mm or more and 3 mm or less.
The power supply device 21b can apply a voltage between the surface electrode 3 and the lower electrode 2 to generate an electric field in the intermediate layer 4. Due to this electric field, a current can be passed through the intermediate layer 4, and electrons can be emitted from the electron emission region 7 of the surface electrode 3. The electrons charge the gas phase component, and charges such as oxygen anions are generated in the gas phase 15. The anion is also applied to the medium 10.

For example, the power supply device 21b, a rectangular wave AC voltage having a frequency 500~10000Hz in crest value 14V _0-p ~24V _0-p can be applied between the surface electrode 3 and the lower electrode 2. Further, the on/off duty ratio can be varied from 1 to 100%. The ion irradiation amount (electron emission amount) from the electron emitting device 20 can be set to 0.5 to 5.0 μA/cm ² .
In addition, the ion irradiation dose during driving is set to a nearly constant value (the fluctuation range is within ±10%, preferably within ±5%). Control can be performed. By setting the ion irradiation dose to a substantially constant value, it is possible to stably advance the culture.

The circular electron-emitting device 20 has a circular shape, a partially lacking circular shape, or an annular shape. For example, the circular electron emitting device 20 can have a shape as shown in FIGS. 1 and 3A to 3C.
The wells 24 and the culture dish of the multi-well plate which is the culture container 23 usually have a circular shape. Therefore, since the circular electron-emitting device 20 has a circular shape, a partially lacking circular shape, or an annular shape, the shape of the circular electron-emitting device 20 can be matched with the shape of the well 24, and a larger electron-emitting device can be obtained. Can be placed in well 24. Further, the opening 6 of the insulating layer 5 is circular or partially circular, and defines the range of the electron emission region 7. Therefore, the shape of the electron emission region 7 can be matched with the shape of the well 24, and electrons can be emitted toward a wider range of the medium 10 in the well 24. As a result, it becomes possible to apply electrical stimulation to more cells 11 cultured in the medium 10. Further, the voltage applied between the lower electrode 2 and the surface electrode 3 can be made relatively small (the current in the device can be made relatively small), the circular electron emission device 20 generates heat, and the cell 11 It is possible to suppress the influence on the culture environment.
Further, since the opening 6 of the insulating layer 5 has a circular shape or a partially lacking circular shape, electrons can be emitted from the electron emitting region 7 with a highly uniform electron emission amount, and the cells 11 cultured in the medium 10 It is possible to give highly uniform electric stimulation to.

  The circular electron-emitting device 20 can be detachably fixed to the base material 13. By arranging the base material 13 on the culture container 23, the surface electrode 3 (electron emission region 7) of the circular electron emission element 20 faces the surface of the medium 10 in the culture container 23 via the gas phase 15. The circular electron-emitting device 20 can be arranged as described above. By arranging the circular electron-emitting device 20 in this way, the gas phase between the circular electron-emitting device 20 and the culture medium 10 is generated by the electrons emitted from the surface electrode 3 (electron-emitting region 7) of the circular electron-emitting device 20. The components of 15 can be charged, and charges such as oxygen anions can be generated in the gas phase 15. Further, the circular electron-emitting device 20 can be arranged so that the surface electrode 3 and the surface of the culture medium 10 are substantially parallel to each other.

The base material 13 has a cylindrical shape. The diameter of the columnar shape can be 0.70 times or more and 0.99 times or less the diameter of the well 24 of the culture container 23. By disposing such a base material 13 in the well 24 so that the upper surface thereof faces the culture medium 10, the upper surface of the base material 13 faces most of the surface of the culture medium 10 in the well 24 via the gas phase 15. can do. Further, the base material 13 can be arranged so as to be concentric with the well 24. Here, the surface of the base material 13 on the lid member 27 side is the lower surface, and the surface of the base material 13 on the medium 10 side is the upper surface.
The circular electron emission device 20 may be detachably fixed to the upper surface of the cylindrical base material 13. The diameter of the circular electron-emitting device 20 can be 0.65 times or more and 1 time or less than the diameter of the upper surface of the base material 13. By arranging the base material 13 to which the circular electron emitting device 20 is fixed in the well 24 so that the circular electron emitting device 20 is on the medium 10 side, the electron emitting region 7 of the circular electron emitting device 20 is well. It is possible to face the surface of the medium 10 in 24 over a wide range through the gas phase 15. This makes it possible to apply electrical stimulation to a wide range of cells 11 in the medium 10.
The base material 13 can have a shape as shown in FIG. 5, for example.
The base material 13 may have a groove on its peripheral surface. As a result, it is possible to suppress the occurrence of leak current.

The material of the base material 13 can be an insulating material. As a result, it is possible to prevent the leak current from flowing. Further, the material of the base material 13 is preferably a PEEK material or a fluorine-based material that is resistant to heat. The member facing the cell 11 is sterilized in advance by using an autoclave. Since it is a treatment with high temperature steam exceeding 150°C, it must withstand repeated thermal deformation of the material.
The base material 13 may be integrated with the lid member 27 or may be fixed to the lid member 27. In this case, the base material 13 can be provided so that the base material 13 projects from the lid member 27 toward the medium 10 side (in the well 24). Further, the circular electron-emitting device 20 can be fixed to the tip end surface of the base material 13 protruding from the lid member 27 so as to be parallel to the surface of the culture medium 10.
The material of the lid member 27 is preferably a PEEK material or a fluorine-based material that is resistant to heat.

The lid member 27 is provided so that the lid member 27 to which the base material 13 and the circular electron-emitting device 20 are fixed can be placed and combined on the culture container 23. Further, the lid member 27 can have an insulating property.
The base material 13, the circular electron-emitting device 20, and the lid member 27 can be provided so that the distance between the surface electrode 3 and the surface of the culture medium 10 is 0.5 mm or more and 3 mm or less. As a result, the anions generated in the gas phase 15 can be easily supplied to the medium 10. The distance between the surface electrode 3 and the surface of the culture medium 10 can be preferably 1 mm or more and 2 mm or less.
For example, the distance between the surface electrode 3 and the surface of the medium 10 can be adjusted by adjusting the depth of the well of the culture container 23, the depth of the medium 10, the height of the substrate 13 protruding from the lid member 27, and the like. Can be adjusted.
By making the circular electron-emitting device 20 face the surface of the culture medium 10 via the vapor phase 15 in this manner, it is possible to prevent the supply of oxygen gas from the vapor phase 15 to the culture medium 10 to be obstructed. Therefore, it becomes possible to maintain the amount of dissolved oxygen in the medium 10.
Further, by fixing the circular electron-emitting device 20 to the base material 13 in a removable manner, the circular electron-emitting device 20 can be easily replaced.

  The cell stimulator 30 may have at least one lower electrode connection terminal 26. The lower electrode connection terminal 26 may be fixed to the base material 13. Further, the lower electrode connection terminal 26 may be a spring pin connector. The spring pin connector has a tube, a spring disposed in the tube, and a pin partially disposed in the tube to push an end of the spring. In the spring pin connector (lower electrode connection terminal 26), the tip of the pin is pressed against the lower electrode 2 by the restoring force of the spring, and the spring pin connector is electrically connected to the lower electrode 2. The lower electrode connection terminals 26a and 26b can be connected to the lower electrode 2 as shown in FIG. 7, for example. The reason for using a plurality of lower electrode connection terminals 26 is that the stimulus to the cells 11 is used under the environment of temperature: 37° C. and relative humidity: 95%, so that the lower electrode 2 has a non-conductive oxide film. This is because the power supply to the lower electrode connection terminal 26 may occur easily, and this risk is reduced.

The cell stimulator 30 can have at least one surface electrode connection terminal 25. The surface electrode connection terminal 25 may be detachably fixed to the base material 13. Further, the surface electrode connection terminal 25 can be provided directly above the insulating layer 5 so as to be in contact with the surface electrode 3. This can prevent the surface electrode 3 from being destroyed. Further, the surface electrode connection terminal 25 may function as a member for fixing the circular electron emission device 20 to the base material 13. The surface electrode connection terminal 25 can be connected to the surface electrode 3 as shown in FIGS. 5 and 7, for example.
The power supply device 21b can apply a voltage between the surface electrode 3 and the lower electrode 2 via the lower electrode connection terminal 26 and the surface electrode connection terminal 25.

The cell stimulator 30 has a charge recovery electrode 8. Further, the cell stimulator 30 may have a plurality of charge recovery electrodes 8.
The charge recovery electrode 8 is provided so that it can contact the medium 10 for culturing the cells 11. Therefore, electrons can flow from the medium 10 to the charge recovery electrode 8, and the potentials of the charge recovery electrode 8 and the medium 10 can be made equal or almost equal. For example, if the charge recovery electrode 8 is grounded, the medium 10 can be prevented from being charged.
In the cell stimulating device 30 of the present embodiment, the charges generated by the circular electron-emitting device 20 are supplied to the medium 10 via the gas phase 15, so that the only electrode contacting the medium 10 is the charge recovery electrode 8. Therefore, electrolysis of water contained in the medium 10 can be suppressed. Further, since the charge recovery electrode 8 has a positive polarity, the mineral components (K ⁺ , Ca ²⁺ , Na ⁺ , Mg ²⁺ etc.) in the medium 10 do not deposit on the charge recovery electrode 8 and the medium 10 It is possible to suppress changes in the component composition.

The charge recovery electrode 8 can be fixed to the base material 13. The charge recovery electrode 8 can be provided so that the charge recovery electrode 8 contacts the culture medium 10 when the lid member 27 is combined with the culture container 23.
The charge recovery electrode 8 may be a pin electrode or a rod electrode. In this case, the charge recovery electrode 8 can be provided so as to project from the base material 13 to the medium 10 side. Further, the charge recovery electrode 8 may be a spring pin connector. In this case, the charge recovery electrode 8 can be provided so that the tip of the charge recovery electrode 8 is pressed against the bottom of the well 24 containing the medium 10.
When the charge recovery electrode 8 is a pin electrode or a rod electrode, the tip of the charge recovery electrode 8 (the portion that contacts the culture medium 10) can have a diameter of 0.1 mm or more and 2 mm or less.

The charge recovery electrode 8 can be provided so as to project from the base material 13 between the end of the upper surface of the base material 13 and the circular electron emission element 20. For example, the charge recovery electrode 8 can be provided as shown in FIGS.
The substrate 13 may have a groove 28 between the charge recovery electrode 8 and the circular electron emission device 20. As a result, it is possible to suppress the occurrence of leak current.
As shown in FIG. 3A, when a part of the circular electron-emitting device 20 has a round shape, the charge collecting electrode 8 projects from the base material 13 in the region where the circle is missing. An electrode 8 can be provided. For example, the charge recovery electrode 8 can be provided as shown in FIG. As a result, the circular electron emitting device 20 arranged on the upper surface of the base material 13 can be enlarged, and the electron emitting region 7 can be enlarged. As a result, electrons can be emitted toward a wider area of the medium 10, and more cells 11 can be electrically stimulated.

  As shown in FIG. 3B, when the circular electron emission device 20 has an annular shape and has the opening 16 at the center, the charge recovery electrode 8 projects from the base material 13 in the opening 16 to recover the charge. An electrode 8 can be provided. For example, the charge recovery electrode 8 can be provided as shown in FIG. As a result, the circular electron emitting device 20 arranged on the upper surface of the base material 13 can be enlarged, and the electron emitting region 7 can be enlarged. As a result, electrons can be emitted toward a wider area of the medium 10, and more cells 11 can be electrically stimulated. Further, the charge recovery electrode 8 can be arranged at the center of the medium 10, and the homogeneity of the electrical stimulation applied to the cells 11 can be improved.

The charge recovery electrode 8 may have a cylindrical shape. In this case, the charge recovery electrode 8 may project from the base material 13 to the medium 10 side so as to surround the circular electron emission element 20. For example, the charge recovery electrode 8 can be provided as shown in FIG. As a result, the charge recovery electrode can be arranged along the inner wall surface of the circular well, and the homogeneity of the electrical stimulation applied to the cells can be improved.
The charge recovery electrode 8 may have a cylindrical shape with a part missing. In this case, the charge recovery electrode 8 may project from the base material 13 to the medium 10 side so as to surround the circular electron emission element 20. For example, the charge recovery electrode 8 can be provided as shown in FIG. As a result, the charge recovery electrode 8 can be arranged along the inner wall surface of the circular well 24, and the homogeneity of the electrical stimulation applied to the cell 11 can be improved. Further, as shown in FIG. 6D, the cell stimulating device 30 may have a plurality of charge recovery electrodes 8.
When the charge recovery electrode 8 has a cylindrical shape or a partially lacking cylindrical shape, the charge recovery electrode 8 can have a thickness of, for example, 0.1 mm or more and 0.5 mm or less.

  The cell stimulator 30 may include an insulator layer 14 that partially covers the charge recovery electrode 8. This insulator layer 14 can be arranged between the circular electron-emitting device 20 and the charge recovery electrode 8. By providing such an insulator layer 14, it is possible to suppress the generation of a leak current between the circular electron emission element 20 and the charge recovery electrode 8. For example, when the charge recovery electrode 8 is a pin electrode or a rod electrode, the insulator layer 14 can be an insulating tube that partially covers the charge recovery electrode 8.

By generating a potential difference between the circular electron emission element 20 and the charge recovery electrode 8 by the power supply device 21a, an electric field is generated in the gas phase 15 between the surface electrode 3 of the circular electron emission element 20 and the surface of the medium 10. Can be made The anions generated by the circular electron emission device 20 emitting electrons along the lines of electric force (gradient of electric field strength) in the electric field of the gas phase 15 can be transported to the surface of the medium 10. It is possible to generate anions (OH ⁻ , Cl ⁻ , O ₂ ^−, etc.) in the medium 10 (current can be passed through the gas phase 15). This anion can give an electrical stimulation to the cells 11 cultured in the medium 10. Further, since the anions move in the medium 10 and flow to the charge recovery electrode 8 (current flows in the medium 10), the medium 10 can be prevented from being charged.

  For example, the charge recovery electrode 8 can be grounded, and a DC voltage of −1000V to −50V can be applied between the circular electron emission element 20 and the ground by the power supply device 21a. The distance between the surface electrode 3 of the circular electron emission device 20 and the surface of the medium 10 can be 0.5 mm to 3 mm (preferably 1 mm or more and 2 mm or less). Since the charge recovery electrode 8 is grounded, the potential of the medium 10 becomes almost 0V, and the potential of the surface electrode 3 becomes -1000V to -50V. Further, since the distance between the medium 10 and the surface electrode 3 is 0.5 mm to 3 mm, it is possible to generate an electric field having a strong electric field intensity in the gas phase 15 between the medium 10 and the surface electrode 3. By using this electric field, the anions generated by the circular electron emission device 20 emitting electrons can be supplied to the medium 10.

The power supply devices 21a and 21b provide a current generated in the gas phase 15 by supplying electric charges from the circular electron-emitting device 20 to the medium 10 and a current generated in the medium 10 by the charge recovery electrode 8 recovering the charges from the medium 10. Can be electrically connected to the circular electron-emitting device 20 and the charge recovery electrode 8 so that the current becomes a loop current.
For example, as shown in FIG. 7, an electric field generated by applying a voltage V _e between the surface electrode 3 and the charge recovery electrode 8 using the power supply device 21 a causes the gas phase 15 to move to the ion A ⁻ (ion (A ⁻ is generated in the gas phase 15 by the electrons emitted from the surface electrode 3) moves to generate a current in the gas phase 15, and the ions A ⁻ reaching the medium 10 are dissolved in the medium 10 and the The medium 10 is stimulated by stimulating the signal transduction system involved in the formation of various elements, and finally moved to the charge recovery electrode 8 as ion B ⁻ (ion B ⁻ is ion A ⁻ itself or another ion species derived from ion A ⁻ ). An electric current is generated. Therefore, a loop current can be passed through a circuit including the power supply device 21a, the surface electrode 3, the gas phase 15, and the medium 10.

  The circular electron-emitting device 20 can surface-emit electrons from the circular electron-emitting region 7 of the surface electrode 3, so that anions are uniformly distributed in the gas phase 15 between the surface of the medium 10 and the electron-emitting region 7. Can be generated. Since this anion can be supplied to the medium 10 by the electric field, the anion can be uniformly generated in the medium 10 located below the electron emitting region 7 of the surface electrode 3 and the electron emitting region 7 Electric stimulation can be uniformly applied to the cells 11 located in the lower part. Therefore, it is possible to uniformly apply electrical stimulation to cells 11 to which electrical stimulation is desired. Further, since the electron emission region 7 has a circular shape, the uniformity of the amount of electron emission in the electron emission region 7 can be improved, and uniform electrical stimulation can be given to the cells 11 located below the electron emission region 7. You can

The electrical stimulation (ion irradiation) by the cell stimulator 30 can be started, for example, at the timing when the seeded cells start cell division.
Ion irradiation of the medium 10 on which the cells 11 are fixed can be performed by setting the irradiation amount, irradiation interval, and irradiation timing suitable for the cell type.
The time for emitting electrons from the circular electron-emitting device 20 (the time for applying voltage by the power supply devices 21a and 21b) can be set to, for example, 5 seconds or more and 1 minute or less, but the purpose of cell type, proliferation, induction of differentiation, etc. Can be changed arbitrarily.
Further, the description of the circular electron emitting device 20 of the first or second embodiment applies to the circular electron emitting device 20 included in the cell stimulating device 30 of the third embodiment unless there is a contradiction.

Fourth Embodiment The fourth embodiment relates to a cell stimulator.
FIG. 8 is a schematic top view of the cell stimulator 30 of this embodiment, and FIG. 9 is a schematic perspective view of the culture container 23. FIG. 10 is a schematic cross-sectional view taken along the broken line BB of the cell culture container 30 in which the cell culture container 30 shown in FIG. 8 and the culture container 23 shown in FIG. 9 are combined so that the broken lines BB coincide with each other.
In the present embodiment, a plurality of electron emitting portions 29a to 29f (these are collectively referred to as electron emitting portions 29) are attached to the lid member 27, and each of the electron emitting portions 29a to 29f includes a charge recovery electrode 8 and a lower portion. The electrode connection terminals 26 (26a to 26f), the surface electrode connection terminals 25 (25a to 25f), the circular electron emission devices 20 (20a to 20f), and the base material 13 (13a to 13f) are provided. The electron emission unit 29 is the cell stimulating device 30 as shown in FIGS. 5 and 6A to 6D. The description of the circular electron emitting device 20 of the first and second embodiments applies to the circular electron emitting device 20 included in the electron emitting portions 29a to 29f unless there is a contradiction. Further, the description of the cell stimulator 30 of the third embodiment applies to the electron emission unit 29 or the cell stimulator 30 of the fourth embodiment unless there is a contradiction.

  The culture container 23 has a plurality of wells 24a to 24f (collectively referred to as wells 24) that accommodate the medium 10 for culturing the cells 11. The cells 11 can be cultured in the medium 10 in each well 24a to 24f. The number of wells 24 included in the culture container 23 is, for example, 6, 12, 18, 24, 32, 48, 96. The well 24 is a circular recess. For example, the culture container 23 shown in FIG. 9 has six wells 24a to 24f.

The lid member 27 has a shape that can be combined with the culture container 23 to cover the well 24. Further, the electron emitting portion 29 is positioned so that the electron emitting portion 29 is located in the well 24 when the lid member 27 is combined with the culture container 23, and the electron emitting region 7 of the surface electrode 3 faces the surface of the culture medium 10. It is fixed to the lid member 27. Further, the electron emitting portion 29 corresponding to each well 24 of the culture container 23 may be attached to the lid member 27. Further, each electron emitting portion 29 may be fixed to the lid member 27 so as to be concentric with the corresponding well 24 when the lid member 27 and the culture vessel 23 are combined.
For example, the cell stimulator 30 shown in FIG. 8 has six electron emitting portions 29a to 29f corresponding to the six wells 24a to 24f of the culture container 23. Further, when the lid member 27 is combined with the culture vessel 23, each electron emitting portion 29a to 29f is located in the well 24 and each circular electron emitting element 20a to 20f is arranged like the cell stimulating device 30 shown in FIG. , Are arranged so as to face the surface of the medium 10 in each well 24.

  2: Lower electrode 3: Surface electrode 4: Intermediate layer 5: Insulating layer 6: Opening 7: Electron emission region 8, 8a to 8f: Charge recovery electrode 10: Medium 11: Cell 13, 13a to 13f: Base material 14: Insulation Body layer 15: Gas phase 16: Opening 20, 20a to 20f: Circular electron-emitting device 21, 21a, 21b: Power supply device 23: Culture vessel 24, 24a to 24f: Well 25, 25a to 25f: Surface electrode connection terminal 26, 26a-26f: Lower electrode connection terminal 27: Lid member 28: Groove 29, 29a-29f: Electron emission part 30: Cell stimulator

Claims (10)

A lower electrode, a surface electrode facing the lower electrode, an intermediate layer arranged between the lower electrode and the surface electrode, and an insulating layer having an opening,
The insulating layer is disposed between the lower electrode and the intermediate layer, or between the intermediate layer and the surface electrode,
The lower electrode, the intermediate layer, and the surface electrode generate a potential difference between the lower electrode and the surface electrode, so that a current flows in the intermediate layer, and electrons are emitted from an electron emission region of the surface electrode. Is provided,
The electron emission region is defined by the opening in the insulating layer,
The circular electron-emitting device is characterized in that the opening has a circular shape or a partially lacking circular shape. The lower electrode, the intermediate layer and the surface electrode are circular, partially lacking circular or annular shape,
The circular electron emission device according to claim 1, wherein the insulating layer has an annular shape surrounding the opening. A circular electron emission device according to claim 1 or 2, and a charge recovery electrode,
The circular electron-emitting device has a circular shape, a partially lacking circular shape or an annular shape, and is provided so as to supply an electric charge to a medium for culturing cells via a gas phase,
The said charge recovery electrode is a cell stimulator provided so that a charge may be collect|recovered from the said culture medium. Further provided with a cylindrical base material,
The cell stimulator according to claim 3, wherein the circular electron-emitting device is detachably fixed on the upper surface of the base material. The circular electron-emitting device has a circular shape with a part cut off,
The cell stimulator according to claim 4, wherein the charge recovery electrode is a pin electrode or a rod electrode and protrudes from the base material in a region where the circular electron emission element is lacking. The circular electron-emitting device has an annular shape,
The cell stimulator according to claim 4, wherein the charge recovery electrode is a pin electrode or a rod electrode, and protrudes from the base material at an opening at the center of the circular electron emission element.   The cell stimulator according to claim 4, wherein the charge recovery electrode has a cylindrical shape or a cylindrical shape with a part thereof cut off, and protrudes from the base material so as to surround the circular electron-emitting device. The charge recovery electrode is an electrode partially covered by an insulating layer,
The cell stimulator according to any one of claims 3 to 7, wherein the insulator layer is arranged between the circular electron-emitting device and the charge recovery electrode. Further equipped with a power supply,
The power supply device is provided so as to generate a potential difference between the lower electrode and the surface electrode, and may generate a potential difference between the circular electron-emitting device and the charge recovery electrode. The cell stimulating device according to any one of claims 3 to 8, which is provided so as to be capable. Further comprising a culture vessel having a circular well for containing the medium,
10. The cell stimulating device according to claim 3, wherein the circular electron-emitting device is arranged so that the surface electrode faces the surface of the medium contained in the well with a gas phase interposed therebetween. ..