JP4692959B2 - Observation substrate and droplet supply device - Google Patents

Description

  The present invention relates to an observation substrate used when observing cells in a liquid in fields related to the life science field, regenerative medicine field, bio-related field, and the like, and a droplet supply device having the observation substrate. .

In recent years, in bio-related research, cells in a solution such as a culture solution are observed in the solution to observe the protein present at the interface between the cells and the solution. One of the important research subjects is to observe changes in proteins when the environment in the liquid is changed instantaneously by exchanging or mixing solutions.
Various methods are known as a method for changing the environment of the solution. As one of the general methods, a method of changing the environment in the liquid by mixing a chemical substance or the like into the solution. It has been known. Specifically, cells are held in a container filled with a solution, and a solution supply pump is connected to the container via a solution supply pipe. And the environment in a liquid is changed by mixing a chemical substance etc. with the syringe etc. from the middle of the solution supply pipe | tube.

As another method, a method is also known in which a syringe or the like is positioned in the vicinity of a cell held in a solution, and the environment around the cell is changed by injecting another solution with the syringe. ing.
In addition, there is also known a method of observing cells in liquid by dropping a droplet-like solution onto cells held on a substrate using a syringe. The environment is changed by dripping.
Then, the environment surrounding the cell is changed by the above-described method, and the change of the protein at this time is scanned with, for example, an optical microscope or a microprobe to obtain an image, information, etc. ( (SPM) and so on.

Here, at present, a technique called liquid dielectrophoresis in which a droplet-like solution is moved along a patterned electrode is known (see, for example, Patent Document 1). This is because when a predetermined high-frequency voltage is applied to a pair of electrode patterns arranged in parallel with a predetermined gap (gap) between them, a droplet-like solution disposed on the electrode pattern is caused to gap by a dielectrophoresis phenomenon. It moves along the electrode pattern in a state where is used as a flow path.
In the future, this technology will be used to supply droplet-like solutions to objects to be supplied, or to easily mix droplet-like solutions together, and apply them to devices in various fields. It has been. For example, application to changing the solution around the cells described above instantaneously is considered.
Japanese translation of PCT publication No. 2002-503047

By the way, the following problems remain in the conventional method.
That is, in the method of injecting the chemical substance and the method of injecting the solution with a syringe, the solution diffuses and it is difficult to instantaneously change the environment of the solution around the cell. In addition, in the method using a syringe, it is necessary to position the syringe itself in the vicinity of the cell that is the observation region regardless of whether injection or dripping is performed. Met. In addition, it is necessary to accurately position the syringe in the vicinity of the cell, but this alignment is difficult and time-consuming. Therefore, it was not possible to perform observation efficiently. Furthermore, in the case of injecting the solution with a syringe, the solution diffuses at the time of injection, so that it is necessary to inject the solution in an amount more than necessary, resulting in high costs.

  The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an observation substrate capable of instantaneously changing a liquid environment around a cell without affecting the observation visual field and the observation. It is providing the droplet supply apparatus provided with the substrate for operation.

In order to achieve the above object, the present invention provides the following means.
The observation substrate of the present invention is provided in a rail shape along the upper surface of the substrate body, with the substrate body having a holding region capable of holding cells at a predetermined position on the upper surface, and one end side positioned in the holding region. An electrode pattern that movably supports the droplet-like solution, and an electrical connection portion that is electrically connected to the electrode pattern and is electrically connectable to the outside. When an AC voltage having a predetermined frequency is applied between the first electrode and the second electrode from the outside via the electrical connection portion, the first electrode and the second electrode arranged in parallel. In addition, the droplet-like solution is moved in any direction along both electrodes by the dielectrophoretic force , and the electrode pattern has a holding portion for temporarily holding the droplet-like solution on the other end side. The two electrodes are each formed in a semicircular shape at the other end. And which, the holder is characterized in that the circular shaped area which the other end of each of the electrodes is combined.

  In the observation substrate according to the present invention, one end side of the rail-like electrode pattern that supports the droplet-like solution, that is, one end side of the first electrode and the second electrode is held in the holding region for holding the cells. positioned. Here, when an alternating voltage of a predetermined frequency is applied between the first electrode and the second electrode from the outside via the electrical connection portion, the liquid droplet solution causes a gap between the two electrodes due to the dielectrophoretic force. It is drawn into the part and moves along both electrodes. Thereby, a droplet-like solution can be supplied to the cells held in the holding area in a timely manner. As a result, the cells are encased in the droplet-like solution in the holding region and are in the liquid. By observing this state, the cells can be observed in the liquid. That is, the holding area is in an observation area state.

In this way, a droplet-like solution can be supplied to cells in a timely manner depending on the electrode pattern, so that the environment around the cell can be instantaneously changed (for example, the environment can be changed by supplying different types of solutions and mixing them). Can do. Therefore, it is possible to clearly observe the change with time of the protein due to this environmental change. At this time, since the solution is in the form of droplets, it directly acts on (influences on) the cells without diffusing as in the prior art.
Further, since the electrode pattern is arranged in a rail shape along the upper surface of the substrate body, there is no obstacle on the upper side of the holding region where the cells are held. That is, it is in a state where it is open to an externally accessible state. Therefore, when the cells in the holding region are observed with, for example, an optical microscope or an SPM probe, there is no influence such as interference with a syringe as in the conventional case. Therefore, it is possible to reliably observe without overlooking changes in the protein over time.

Further, since the solution in the form of droplets is supplied, the solution is not wasted, and a necessary minimum amount can be used, and the cost can be reduced.
In addition, long-term observation can be performed while preventing the cells from drying by supplying the same kind of solution in a timely manner. At this time, long-term observation can be performed with the concentration kept constant by using a buffer solution. In addition, the electrode pattern is not always applied, but is applied only when the solution is moved, so that an increase in temperature can be prevented as much as possible. Therefore, it is difficult to affect the solution and cells with heat.
Furthermore, since the holding part is provided on the other end side of the electrode pattern, the solution can be reliably held in the form of droplets even when no AC voltage is applied to the electrode pattern. Therefore, when the alternating voltage is applied, the droplet-like solution can be quickly moved toward the cells.

  The observation substrate of the present invention is characterized in that, in the observation substrate of the present invention, a plurality of the electrode patterns are provided radially around the holding region.

In the observation substrate according to the present invention, since a plurality of electrode patterns are provided in a radial pattern, various solutions can be instantaneously and efficiently mixed into cells, and various observations can be made under various combinations. It can be carried out.
In addition, since a plurality of electrode patterns are provided, it is possible to first move the solution enclosing the cells before supplying a new solution, and then supply a new solution. That is, the solution can be exchanged instantaneously. As a result, the environment around the cell can be further changed, and the change with time of the protein can be clearly observed.

  The observation substrate of the present invention is characterized in that, in the observation substrate of the present invention, the electrode pattern is branched into a plurality on the way from the one end side to the other end side.

  In the observation substrate according to the present invention, since the electrode pattern is branched into a plurality of parts on the way, for example, after different types of solutions are reacted (mixed) once at the branch points, this reaction solution is supplied to the cells. can do. Therefore, more diversified observation can be performed.

  Further, the observation substrate according to the present invention is the observation substrate according to any one of the above-described present invention, wherein the upper surface of the substrate main body is hydrophobic to repel the droplet-like solution so as to surround the holding region. A characteristic film is formed.

  In the observation substrate according to the present invention, since the hydrophobic film is formed so as to surround the periphery of the holding region, even if a part of the solution splashes around when the solution is supplied to the cells. , Bounced off the hydrophobic membrane and returned to the holding region. Therefore, a predetermined amount of solution can be reliably supplied to the cells.

  The observation substrate according to the present invention is characterized in that, in the observation substrate according to the present invention, the hydrophobic film is formed so as to surround the electrode pattern.

  In the observation substrate according to the present invention, since the hydrophobic film is formed so as to surround the periphery of the electrode pattern, when the solution moves on the electrode pattern, a part of the solution is temporarily out of the electrode pattern. Even if splashed, it is repelled by the hydrophobic membrane. Therefore, a predetermined amount of solution can be reliably supplied to the cells.

  An observation substrate according to the present invention is the observation substrate according to any one of the present invention described above, wherein the electrode pattern is coated with a thin protective film.

  In the observation substrate according to the present invention, since the electrode pattern is coated (coated) with a protective film, direct contact with the outside can be prevented. Therefore, the electrode pattern is protected, and durability and reliability can be improved.

  In addition, a droplet supply device of the present invention is a voltage application unit that is electrically connected to the observation substrate according to any one of the present invention and the electrical connection portion, and applies an alternating voltage of a predetermined frequency to the electrode pattern. And control means for controlling the operation of the voltage application means.

  In the droplet supply device according to the present invention, the control means operates the voltage application means to apply an alternating voltage of a predetermined frequency to the electrode pattern, thereby moving the solution in a direction toward the cell or away from the cell. The environment can be changed instantaneously by supplying or exchanging the solution. The solution may be manually dropped on the electrode pattern, for example.

  Further, the droplet supply device of the present invention is the above-described droplet supply device of the present invention, wherein the droplet supply solution is dropped on the electrode pattern, and the solution supply means whose operation is controlled by the control means is provided. It is characterized by having.

  In the droplet supply device according to the present invention, since the solution supply means is provided, the solution can be dropped on the electrode pattern when necessary without manpower, and the control means drops the solution and moves the solution. It is easy to use and is very convenient.

According to the observation substrate according to the present invention, since the droplet-like solution can be supplied to the cells in a timely manner by the electrode pattern, the environment around the cells can be instantaneously changed. Therefore, it is possible to clearly observe changes in the protein over time. In addition, since there are no obstacles around the holding area, the cells in the holding area can be reliably observed without any influence by an optical microscope, an SPM probe, or the like. Therefore, it is possible to reliably observe without overlooking changes in protein over time.
Further, since the solution in the form of droplets is supplied, the required amount can be reduced without wasting the solution, and the cost can be reduced.

  Further, according to the droplet supply device of the present invention, the solution can be moved in a direction toward the cell or in a direction away from the cell, and the environment around the cell can be instantaneously changed by supplying or exchanging the solution. Can be changed.

Hereinafter, an embodiment of an observation substrate and a droplet supply device according to the present invention will be described with reference to FIGS.
As shown in FIG. 1, the droplet supply device 1 of the present embodiment is electrically connected to an observation substrate 2 on which cells A are placed and an electrical connection portion 12 described later of the observation substrate 2, which will be described later. A driving power supply (voltage applying means) 3 for applying an AC voltage of a predetermined frequency (for example, a frequency of 300 kHz, an AC voltage of 500 Vrms) to the electrode pattern 11, and a controller (control means) 4 for controlling the operation of the driving power supply 3. It has.

  In addition, the observation substrate 2 of this embodiment is placed on the stage 6 of the optical microscope 5. The optical microscope 5 includes an objective lens 7 disposed above the stage 6, an eyepiece 8 for observing the cell A placed on the observation substrate 2 through the objective lens 7 in the liquid, and the stage 6. It is an upright microscope provided with an illumination system 9 that irradiates illumination light toward the observation substrate 2 from below.

  As shown in FIGS. 2 to 4, the observation substrate 2 has a glass substrate (substrate body) 10 having a holding region S capable of holding cells A at a predetermined position on the upper surface, and one end side in the holding region S. In this state, the electrode pattern 11 is provided in a rail shape along the upper surface of the glass substrate 10 and movably supports the droplet-like solution W, and is electrically connected to the electrode pattern 11 and can be electrically connected to the outside. The electrical connection part 12 is provided.

Four (plural) of the electrode patterns 11 are provided radially with the holding region S as the center. That is, the electrode pattern 11 is formed in a straight line so as to be orthogonal to each side of the glass substrate 10 at an angle of 90 degrees with the holding region S as the center. That is, when the glass substrate 10 is viewed from above, the four electrode patterns 11 are combined to form a cross shape.
In the present embodiment, a liquid reservoir (holding portion) 11 a that temporarily holds the droplet-like solution W is formed on the other end side of the electrode pattern 11.
Each electrode pattern 11 includes a first electrode 20 and a second electrode 21 that are arranged in parallel with a predetermined interval therebetween, and has a predetermined frequency from the driving power supply 3 via the electrical connection portion 12. When an AC voltage is applied, the droplet-like solution W can be moved in any direction along the electrodes 20 and 21 by the dielectrophoretic force.
The other electrodes 20 and 21 are formed in a semicircular shape at the other end, and a combination of these electrodes forms a circular droplet forming region. That is, this region is the liquid reservoir 11a, and the solution W can be held in the form of droplets using the surface tension.
The electrode pattern 11 is coated with a thin protective film (for example, an insulating film of SiO ₂ ) 13 so that it is not exposed to the outside.

Next, a method for manufacturing the observation substrate 2 will be described below.
First, as shown in FIG. 5, a metal layer 15 is formed by metal-coating a metal material to be the electrode pattern 11 on the entire upper surface of the glass substrate 10 by, for example, vapor deposition or sputtering. Next, as shown in FIG. 6, a photoresist film 16 is patterned on the upper surface of the metal layer 15 by a lithography technique so as to be the electrode pattern 11 shown in FIG.
Next, as shown in FIG. 7, the metal layer 15 is etched using the photoresist film 16 as a mask to produce the electrode pattern 11. After the etching process, as shown in FIG. 8, by removing the photoresist film 16 used as a mask, the electrode pattern 11 according to the patterning of the photoresist film 16 can be formed on the upper surface of the glass substrate 10. The photoresist film 16 may be a positive type or a negative type.
Finally, as shown in FIG. 9, a protective film 13 is formed in a thin film on the entire upper surface of the glass substrate 10 by, for example, sputtering or CVD (chemical vapor deposition). Thereby, the observation substrate 2 in which the electrode pattern 11 is coated with the protective film 13 can be manufactured.

The case where the time-dependent change of the cell A is observed with the optical microscope 5 using the observation substrate 2 and the droplet supply device 1 configured as described above will be described below.
First, the cells A are held in the holding region S of the observation substrate 2, and a desired solution W is dropped into the liquid reservoir 11a with a syringe or the like, for example. At this time, since the liquid reservoir 11a has a circular shape, the dropped solution W is reliably held in a droplet state by its own surface tension.
Next, the observation substrate 2 is placed on the stage 6 of the optical microscope 5, and the position of the stage 6 is adjusted so that the holding area S is within the observation area of the objective lens 7.

  Next, the driving power source 3 is operated by the controller 4 to apply an AC voltage having a predetermined frequency between the first electrode 20 and the second electrode 21 of the electrode pattern 11 on which the solution W is placed. By this voltage application, the solution W held in the liquid reservoir 11a is drawn into the gap between the first electrode 20 and the second electrode 21 by the dielectrophoretic force, and as shown in FIG. Move up towards cell A. And the solution W which moved to the one end side of the electrode pattern 11 contacts with the cell A currently hold | maintained at the holding | maintenance area | region S, as shown in FIG. As a result, as shown in FIG. 12, the cell A is stabilized in the encapsulated state. That is, the cell A is in a state of being held in the holding region S in a state of being present in the liquid. Therefore, the cell A can be observed in the liquid through the objective lens 7.

Here, observation of changes over time when the environment surrounding the cell A is changed by mixing the solution W with another type of solution W ′ (for example, solutions having different concentrations, ions, and temperatures). Do.
First, another type of solution W ′ is dropped on the liquid reservoir 11a of any one of the electrode patterns 11 with, for example, a syringe 17 or the like. Then, the driving power source 3 is operated by the controller 4 to apply an AC voltage having a predetermined frequency to the electrode pattern 11 on which the solution W ′ is placed. By this voltage application, the other solution W ′ moves on the electrode pattern 11 and is supplied to the cell A in the same manner as described above.
By this supply, different types of solutions W and W ′ can be mixed, and the environment around the cell A can be instantaneously changed. Therefore, the observer can observe the change with time of the protein around the cell A due to the environmental change through the objective lens 7 and the eyepiece lens 8. At this time, since the solutions W and W ′ are in the form of droplets, they directly act (influence) on the cells A without diffusing as in the prior art.

In addition, since the electrode pattern 11 is provided in a rail shape along the upper surface of the glass substrate 10, there is no obstacle around the holding region S where the cells A are held (upper side). Yes. That is, it is in a state where it is open to an externally accessible state. Therefore, the syringe is not affected by interference or the like as in the prior art, and the objective lens 7 can be positioned in the vicinity of the cell A in the holding region S, so that the field of view to be observed is not hindered. Therefore, it is possible to reliably observe without overlooking changes in the protein over time.
Further, since the droplet-like solutions W and W ′ are supplied, the solution W and W ′ can be used in the minimum amount without being wasted, and the cost can be reduced.

Further, by supplying the same type of solution W in a timely manner, long-term observation can be performed while preventing the cells A from drying. At this time, long-term observation can be performed with the concentration kept constant by using a buffer solution. In addition, the electrode pattern 11 is not always applied, but is applied only when the solution W is moved, so that the temperature rise of the solution W can be prevented as much as possible. Therefore, it is difficult to affect the cells A with heat.
In addition, since the electrode pattern 11 is provided with a liquid reservoir 11a, when an alternating voltage is applied, the droplet-like solution W can be quickly supplied to the cells A, and the observation is smoothly performed. It can be carried out.
Further, since the electrode pattern 11 is coated (coated) with the protective film 13, direct contact with the outside can be prevented. Therefore, the electrode pattern 11 is protected and durability and reliability can be improved.

Furthermore, since the observation substrate 2 of this embodiment is provided with a plurality of electrode patterns 11, it is also possible to perform solution exchange.
That is, before supplying the new solution W ′ to the cell A, the existing solution W enclosing the cell A is moved to the electrode pattern 11 other than the electrode pattern 11 on which the new solution W ′ is placed, Thereafter, a new solution W ′ can be supplied to the cell A.
This solution exchange will be specifically described below with reference to FIGS.

First, as shown in FIG. 13, for example, the same type of solution W is moved using the dielectrophoretic force toward the cells A held in the holding region S in a state of being wrapped in the solution W. This state is a state in which the solution W enclosing the cell A is not in contact with the electrode pattern 11.
Next, as shown in FIG. 14, the solution W that has moved to the other end side of the electrode pattern 11 comes into contact with the solution W that encloses the cells A and is drawn by surface tension. As a result, as shown in FIG. 15, the amount of the solution W enclosing the cells A increases, and the solution W comes into contact with one end side of the plurality of electrode patterns 11.

  In this state, an alternating voltage of a predetermined frequency is applied to an electrode pattern 11 different from the one in which the solution W is moved. As a result, as shown in FIG. 16, the solution W having an increased amount of liquid is drawn into the electrode pattern 11 to which the AC voltage is applied by the dielectrophoretic force, and the liquid is applied on the electrode pattern 11 as shown in FIG. Move in drops. Thereby, the solution W does not exist around the cell A.

In addition, when the above-described solution W is detached, as shown in FIG. 17, an alternating voltage having a predetermined frequency is simultaneously applied to the electrode pattern 11 so that a new solution W ′ is obtained by using the dielectrophoretic force. Move toward A. As a result, as shown in FIG. 18, a new solution W ′ can be supplied to the cell A, and the cell A can be wrapped with the new solution W ′.
By passing through each procedure mentioned above, solution exchange can be performed instantly. Therefore, the environment around the cell A can be changed more, and the change with time of the protein can be observed more clearly.

  The technical scope of the present invention is not limited to the above embodiment, and various modifications can be made without departing from the spirit of the present invention.

For example, in the above embodiment, the cell A is observed in the liquid using the upright optical microscope 5 in which the objective lens 7 is disposed above the stage 6 and the illumination system 9 is disposed below the stage 6. Observation may be performed with an inverted optical microscope in which the illumination system 9 is disposed above and the objective lens 7 is disposed below the stage 6.
Further, the observation is not limited to the optical microscope 5, and for example, observation may be performed using a scanning probe microscope such as an atomic force microscope having a probe at the tip of a cantilever. Even in this case, since the periphery of the holding region S is open, the probe can be positioned in the vicinity of the cell A without any obstacle, and the change over time of the protein can be reliably detected. Can be observed.

Moreover, in the said embodiment, although it was set as the structure which provided the four electrode patterns 11, it is not restricted to four, One may be sufficient and two or more two or more may be provided. Further, the electrode pattern 11 may be provided so as to be branched into a plurality on the way from the one end side located in the holding region S to the other end side.
By carrying out like this, after reacting (mixing) a different kind of solution once at a branch point, this reacted solution can be supplied to the cell A. Therefore, more diversified observation can be performed.

Further, a hydrophobic film that repels the solution W may be formed on the upper surface of the glass substrate 10 so as to surround the holding region S. By doing so, when supplying the solution W to the cells A, even if a part of the solution W splashes around, it is repelled by the hydrophobic film and returns to the holding region S. Therefore, a predetermined amount of the solution W can be reliably supplied to the cells A.
Further, this hydrophobic film may be formed so as to surround the electrode pattern 11. By doing so, when the solution W is moving on the electrode pattern 11, even if a part of the solution W splashes outside the electrode pattern 11, it is repelled by the hydrophobic film. Therefore, a predetermined amount of the solution W can be supplied to the cells A more reliably.

Further, a solution supply means for dropping the droplet-like solution W onto the electrode pattern 11 may be added to the droplet supply apparatus 1 of the above embodiment, and the controller 4 may be configured to control the operation of the solution supply means. I do not care.
By doing in this way, the solution W can be dripped on the electrode pattern 11 as needed, without manpower. Therefore, the dropping of the solution W and the movement of the solution W can be automatically controlled by the controller 4, and an easy-to-use and excellent convenience can be realized.

It is a block diagram which shows one Embodiment of the droplet supply apparatus which concerns on this invention. It is a top view which shows one Embodiment of the board | substrate for observation which concerns on this invention. FIG. 3 is an enlarged view around a holding region of the observation substrate shown in FIG. 2. It is a cross-sectional arrow XX figure of the board | substrate for observation shown in FIG. It is a figure which shows the manufacturing process of the board | substrate for observation shown in FIG. 2, Comprising: It is a figure which shows the state which formed the metal layer in the whole upper surface of a glass substrate. It is a figure which shows the manufacturing process of the board | substrate for observation shown in FIG. 2, Comprising: It is a figure which shows the state which patterned the photoresist film in the shape of the electrode pattern from the state shown in FIG. FIG. 7 is a diagram illustrating a manufacturing process of the observation substrate illustrated in FIG. 2, and illustrates a state in which an electrode pattern is formed by etching a metal layer using a photoresist film as a mask from the state illustrated in FIG. 6. It is a figure which shows the manufacturing process of the board | substrate for observation shown in FIG. 2, Comprising: It is a figure which shows the state which removed the photoresist film from the state shown in FIG. It is a figure which shows the manufacturing process of the board | substrate for observation shown in FIG. 2, Comprising: It is a figure which shows the state which formed the thin film-like protective film from the state shown in FIG. 8, and coat | covered the electrode pattern. It is sectional drawing of the board | substrate for observation which shows the state which has moved the droplet-form solution on an electrode pattern toward the cell currently hold | maintained on the holding | maintenance area | region by the dielectrophoretic force. It is sectional drawing of the board | substrate for observation which shows the state from which the solution contacted the cell from the state shown in FIG. 10, and was drawn into the cell by surface tension. It is sectional drawing of the board | substrate for observation which shows the state which the solution wrapped the cell from the state shown in FIG. 11, and made this cell exist in a liquid. It is a figure which shows the process at the time of exchanging the solution which wraps a cell, Comprising: The state where the same kind of solution is moved by the dielectrophoretic force along the electrode pattern toward the cell wrapped in the solution It is sectional drawing of the board | substrate for observation shown. It is a figure which shows the process at the time of replacing | exchanging the solution which wraps the cell, Comprising: The state which is attracted | sucked by the surface tension while the solution which has moved from the state shown in FIG. It is sectional drawing of the board | substrate for observation shown. It is a figure which shows the process at the time of replacing | exchanging the solution which wraps the cell, Comprising: The solution amount which moved is taken in from the state shown in FIG. It is sectional drawing of the board | substrate for observation which shows the state which has contacted the one end side of the electrode pattern. FIG. 16 is a diagram illustrating a process for exchanging a solution enclosing a cell, and is a cross-sectional view of an observation substrate illustrating a state in which the solution enclosing the cell is moving on the electrode pattern from the state illustrated in FIG. 15. It is. It is a figure which shows the process at the time of replacing | exchanging the solution which wraps the cell, Comprising: The solution which wraps the cell moves on an electrode pattern from the state shown in FIG. It is sectional drawing of the board | substrate for observation which shows the state which the solution is moving toward the cell on an electrode pattern. It is a figure which shows the process at the time of replacing | exchanging the solution which wraps the cell, Comprising: From the state shown in FIG. 17, the new solution contacts the cell and shows the state which exchange of the solution was completed after wrapping the cell. It is sectional drawing of the board | substrate for observation.

Explanation of symbols

A cell S holding area 1 droplet supply device 2 substrate for observation 3 power supply for driving (voltage applying means)
4 Controller (control means)
10 Glass substrate (substrate body)
11 Electrode pattern 11a Liquid reservoir (holding part)
12 Electrical connection part 13 Protective film 20 1st electrode 21 2nd electrode

Claims (8)

A substrate body having a holding region capable of holding cells at a predetermined position on the upper surface;
An electrode pattern that is provided in a rail shape along the upper surface of the substrate body with one end side located in the holding region, and supports the liquid solution in a movable manner,
An electrical connection portion electrically connected to the electrode pattern and electrically connectable to the outside;
The electrode pattern includes a first electrode and a second electrode arranged in parallel with a predetermined interval therebetween, and the first electrode and the second electrode are externally connected via the electrical connection portion. When an alternating voltage of a predetermined frequency is applied between them, the droplet-like solution is moved in any direction along both electrodes by dielectrophoretic force ,
The electrode pattern has a holding part for temporarily holding the liquid droplet solution on the other end side,
Each of the electrodes has a semicircular shape at the other end,
The observation substrate according to claim 1, wherein the holding portion is a circular region formed by combining the other end sides of the electrodes . The observation substrate according to claim 1,
An observation substrate, wherein a plurality of the electrode patterns are provided radially around the holding region. The observation substrate according to claim 1 or 2,
The observation substrate according to claim 1, wherein the electrode pattern is branched into a plurality on the way from the one end side to the other end side. In the observation substrate according to any one of claims 1 to 3 ,
An observation substrate, wherein a hydrophobic film that repels the solution is formed on an upper surface of the substrate body so as to surround the periphery of the holding region. The observation substrate according to claim 4 ,
The substrate for observation, wherein the hydrophobic film is formed so as to surround the electrode pattern. In the observation substrate according to any one of claims 1 to 5 ,
An observation substrate, wherein the electrode pattern is coated with a thin protective film. The observation substrate according to any one of claims 1 to 6 ,
A voltage applying means that is electrically connected to the electrical connecting portion and applies an alternating voltage of a predetermined frequency to the electrode pattern;
And a control means for controlling the operation of the voltage application means. The droplet supply device according to claim 7 , wherein
A droplet supply device comprising a solution supply unit that drops the droplet-like solution onto the electrode pattern and whose operation is controlled by the control unit.

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