JP2004000163A - Cell used for treating cell - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To trap a cell at a specific place in a microchip by a simple means. <P>SOLUTION: A minute flow channel groove 6 used as a flow channel for a liquid sample having a width and a depth of several hundred mm at most and holes 3 and 4 for introducing and discharging the sample are formed on one surface of a glass substrate 1. A cell culture chamber 5 comprised of a circular concave part having a depth almost equal to that of the flow channel and a diameter of about 1 mm is formed on a glass substrate 2. The surface of the substrate 1 where the flow channel groove 6 is formed and the surface of the substrate 2 where the cell culture chamber 5 is formed are faced to each other and adhered to be joined in a liquidtight manner. Thus, when a solution containing a cell is poured from the introduction hole 3, the solution flows along the flow channel groove 6 and the cell is let to flow toward the discharge hole 4 to be trapped in the cell culture chamber 5. <P>COPYRIGHT: (C)2004,JPO

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
INDUSTRIAL APPLICABILITY The present invention is an effective means for handling cells, and particularly relates to a biotechnology research using a microchip.
[0002]
[Prior art]
BACKGROUND ART In the fields of drug discovery, food, crop development, genetic engineering, and the like, research dealing with cells is widely performed. Conventionally, cells have been cultured in a culture place or the like placed in a petri dish or the like and used for various experiments. In order to elucidate the function of cells, it is necessary to examine the reactions caused by many drugs and introduced substances. Therefore, there is a demand for a method in which a specific reaction can be efficiently performed, and a result can be obtained with a large number of samples at a time and with a small amount of samples.
[0003]
In recent years, in the field of analytical chemistry, μTAS (Micro Total Analysis Systems) has been actively studied, and it is expected that a microchip is used to speed up analysis, reduce samples, and save solvents. It has been shown that a reaction in a micro space on a microchip can improve the reaction efficiency as compared with a reaction using a conventional chemical operation.
[0004]
[Problems to be solved by the invention]
However, in a normal microchannel, there is a problem that the cells do not adhere to the wall of the channel due to the flow of the solution, and it is difficult to culture the cells. There is also a method that promotes cell adhesion by modifying the chemical properties of the inner surface of the flow channel.However, in this case, the entire inner wall of the flow channel is modified, so cells must be cultured only at the intended location. Is difficult or difficult to apply when manufacturing a microchip having a complicated function.
An object of the present invention is to provide a cell in which cells can be trapped at a specific place in a microchip by a simple means and cell culture or the like can be performed at this place.
[0005]
[Means for Solving the Problems]
In the channel in the microchip, the solution is always flowing due to a slight difference in the liquid level. For this reason, the cells introduced into the flow channel are distributed at various positions in the flow channel, and the cells are not activated because they cannot be properly attached to the wall surface.
The microchip to which the present invention is directed refers to a microchip in which a flow path and other parts necessary for the reaction and treatment of a solution are formed on a substrate, and the size is not particularly limited.
[0006]
The present invention provides a cell culture chamber having a step in the flow path, and a cell having a structure in which the cross-sectional area is sufficiently large with respect to the cross-sectional area of the flow path. In this method, the cells can be easily trapped in the cell culture room by suppressing them, and processing such as cell culture in a microchip can be performed.
[0007]
That is, the cell of the present invention is provided with a flow path formed inside the base and through which liquid can flow, a liquid inlet and outlet connected to both ends of the flow path, and provided in the flow path inside the base, On the other hand, the base is integrally provided with a cell culture chamber having a step and having a cross-sectional area perpendicular to the flow direction of the liquid larger than the cross-sectional area of the flow path.
[0008]
[Action]
Since the cell culture chamber has a large capacity with respect to the flow path, the flow of the solution in the cell culture chamber is slow. Therefore, cells can be trapped in the cell culture room.
In addition, since the cell culture chamber has a step that is depressed downward or upward with respect to the flow path, the cells settled at the bottom of the cell culture chamber and the vicinity of the floating cells become a quiet environment and the cells easily adhere to the wall surface .
[0009]
BEST MODE FOR CARRYING OUT THE INVENTION
In order to increase the capacity of the cell culture chamber with respect to the flow path, the depth of the step and the bottom area of the lower surface or the upper surface of the cell culture chamber may be increased. For example, the depth of the step in the cell culture chamber is preferably larger than the depth of the flow path.
[0010]
Examples of the material of the base include a glass substrate such as a synthetic quartz substrate and a Pyrex (registered trademark of Corning Glass Works (USA)) glass substrate, a silicon substrate, and a plastic having chemical resistance.
In order to form a flow channel, a cell culture chamber, and a through hole in such a substrate, a micro-machining technology based on a semiconductor manufacturing technology can be used.
[0011]
In order to capture the cells in the cell culture chamber, it is preferable that the inner surface of the cell culture chamber is subjected to a chemical modification for enhancing the adhesion of the cells. As such a chemical modification, a method conventionally applied to a culture vessel can be used, and examples thereof include a collagen coat, a poly-L-lysine coat, and a gelatin coat.
[0012]
One example of a method for manufacturing the cell of the present invention is to bond a substrate having a flow path or the like formed on a bonding surface.
Since it is possible to process both of the two substrates to be bonded, for example, a flow path is formed in a substrate serving as a cover, and a cell culture chamber is formed in a substrate serving as a base, and then these substrates are bonded. Thereby, the cell of the present invention can be manufactured. In the cell thus produced, the base consists of two substrates, the upper substrate has a channel formed on the surface as a groove having a bottom, and the lower substrate has a cell culture chamber on the surface. It is formed as a recess, and both substrates are joined so that the flow path and the cell culture chamber are on the inside, and a liquid port is provided on the upper substrate. In this embodiment, the bottom surface of the cell culture chamber is lower than the flow channel.
[0013]
In the cell of this mode, in the processing step of forming a flow channel and a concave portion for the cell culture chamber on the substrate, the flow channel is formed on one substrate, and the concave portion for the cell culture room is formed on the other substrate. Therefore, it is only necessary to form a groove or a recess having a constant depth in each substrate, and processing is easy.
[0014]
In addition, both the flow path and the cell culture chamber can be formed on one of the two substrates to be joined. The cell manufactured by forming both the flow path and the cell culture chamber on the lower substrate has the above-mentioned liquid inlet / outlet provided on the upper substrate, and the lower substrate has the same surface on the same surface as the groove having the bottom on the flow path. And the cell culture chamber is formed as a concave portion having a deeper bottom than the flow path, and both substrates are joined so that the flow path and the cell culture chamber are inside. Also in this embodiment, the bottom surface of the cell culture chamber is at a position lower than the flow channel.
[0015]
The cell manufactured by forming both the flow path and the cell culture chamber on the upper substrate has the same surface on the upper substrate, the flow path is formed as a groove having a bottom, and the cell culture chamber has a deeper bottom than the flow path. The lower substrate is a substrate with a flat surface, the flow path, the cell culture chamber, and the two substrates are joined such that the flat surface of the lower substrate is inside, and the upper substrate is Is provided with a liquid inlet / outlet. In this embodiment, the bottom surface of the cell culture chamber is located higher than the flow channel.
[0016]
In these embodiments, since the bottom or top surface of the cell culture chamber is formed at a height different from the flow path, chemical modification to capture cells only on the bottom or top surface of the cell culture room is performed. Preferably, the channel is not subjected to the chemical modification.
[0017]
The cell of the present invention can also be manufactured by bonding three substrates. In one embodiment of such a cell of the present invention, a channel is formed as a groove having a bottom on the surface of the uppermost substrate, a cell culture chamber is formed as a through hole in the intermediate substrate, and the lowermost substrate is flat. These three substrates are joined so that the intermediate substrate is interposed therebetween, and the flow path forming surface of the uppermost substrate and the flat surface of the lowermost substrate are on the inner side. The substrate is provided with a liquid inlet / outlet. In this configuration, the cell culture chamber is located lower than the flow channel.
[0018]
In another type of cell formed by laminating three substrates, a liquid inlet / outlet is provided on the uppermost substrate, a channel is formed as a groove having a bottom on the surface of the intermediate substrate, and a cell culture chamber penetrates. The lowermost substrate is formed as a hole, and the lowermost substrate is a substrate having a flat surface. These three substrates sandwich the intermediate substrate, the flow path forming surface of the intermediate substrate faces the uppermost substrate, and the lowermost substrate is Are joined so that the flat surface of the inside faces inside. Also in this mode, the cell culture chamber is located at a position lower than the flow channel.
[0019]
Still another form of a cell formed by bonding three substrates is a substrate in which the uppermost substrate has a flat surface, and the intermediate substrate has a channel formed as a groove having a bottom on the surface thereof. The chamber is formed as a through hole, and the lowermost substrate is also a substrate having a flat surface. These three substrates sandwich the intermediate substrate, and the flow path forming surface of the intermediate substrate is on the flat surface of the lowermost substrate. They are joined so that the flat surface of the uppermost substrate faces the inside so as to close the through hole of the cell culture chamber, and the intermediate substrate is provided with the liquid inlet / outlet. In this configuration, the cell culture chamber is located higher than the flow channel.
[0020]
Still another type of cell manufactured by bonding three substrates is a substrate in which the uppermost substrate has a flat surface, a cell culture chamber is formed as a through-hole in the intermediate substrate, and a cell culture chamber is formed on the surface of the lowermost substrate. Is formed as a groove having a channel with a bottom. These three substrates sandwich the intermediate substrate, the channel forming surface of the lowermost substrate is on the inside, and the flat surface of the uppermost substrate is on the inside. It is joined so as to close the through hole of the culture chamber, and a liquid port is provided in the intermediate substrate. Also in this mode, the cell culture chamber is located at a position higher than the flow channel.
[0021]
In these cells prepared by laminating three substrates, the surface opposite to the surface where the flow path is present is bonded to the intermediate substrate and the surface of the substrate which is to be the inner surface of the cell culture chamber is The chemical modification that enhances the cell adhesion as exemplified in the example has been performed, or the surface opposite to the surface where the flow path is present is bonded to the intermediate substrate on the surface opposite to the intermediate substrate as the substrate constituting the cell culture chamber. It is preferable to use a material having a high cell adhesion. As the substrate made of a material having a high cell adhesion, for example, a plastic material such as polystyrene, a glass plate having a controlled surface state, or the like can be used.
[0022]
Further, in these cells manufactured by bonding three substrates, the thickness of the substrate that is joined to the intermediate substrate on the surface opposite to the surface where the flow path exists and constitutes the cell culture chamber is optically The thickness is preferably set to a thickness suitable for microscopic observation, for example, about 0.15 mm.
[0023]
In the case of these types of cells in which the base is composed of three substrates, the depth of the step in the cell culture chamber is determined by the thickness of the intermediate substrate. The depth of the step in the cell culture chamber can be set arbitrarily by adjusting the thickness of the cell culture chamber.
[0024]
Hereinafter, the present invention will be described more specifically with reference to the drawings.
FIG. 1 shows a first embodiment of a cell realized by the present invention, in which two substrates are joined. (A1) is a plan view of a surface outside the upper substrate 1, (A2) is a plan view of a surface inside the same substrate 1, (B1) is a plan view of a surface inside the lower substrate 2, B2) is a plan view of a surface outside the substrate 2, (C1) is a plan view of a completed cell, (C2) is a cross-sectional view of the cell at the XX line position, and (C3) is a view of the cell. It is sectional drawing in the YY line position.
[0025]
In this figure, reference numerals 1 and 2 denote glass substrates, for example, synthetic quartz substrates. On one surface of the glass substrate 1, there are formed minute flow channel grooves 6 having a width and depth of several hundreds μm or less and used as liquid sample flow channels, and liquid inlet / outlet holes 3 and 4 for sample introduction and discharge. I have.
[0026]
On the other hand, a cell culture chamber 5 is formed in the glass substrate 2 and has a depth approximately the same as that of the flow channel and a circular recess having a diameter of about 1 mm.
The surface of the substrate 1 on which the flow channel 6 is formed and the surface of the substrate 2 on which the cell culture chamber 5 is formed are brought into close contact with each other and joined in a liquid-tight manner. A chip type cell having a cell culture chamber 5 is formed.
As described in a later manufacturing method, a method such as bonding with a hydrofluoric acid solution can be used for bonding the glass substrates.
[0027]
In the embodiment, the shape of the through holes 3 and 4 is tapered so as to become wider as it goes upward, so that the injection and discharge of the sample solution is facilitated. However, the shape of the through holes 3 and 4 is not limited to such a tapered shape, and may be a hole having a vertical wall surface. The same applies to the following embodiments.
[0028]
In the cell having such a configuration, the substrate 1 is arranged on the upper side and the substrate 2 is arranged on the lower side, and when a solution containing cells is poured from the introduction hole 3, the solution flows along the channel groove 6, Flows in the direction of the discharge hole 4. At this time, since the flow rate in the cell culture chamber 5 is significantly reduced, and the cell culture chamber 5 is disposed below the flow channel 6, the cells are trapped in the cell culture chamber 5. The cell culture chamber 5 exists below the bottom surface of the flow channel 6, and since the flow rate of the liquid in the cell culture chamber 5 is reduced, the cells submerged at the bottom of the cell culture chamber 5 A flowing force is not easily applied, and adhesion to the bottom surface is promoted.
[0029]
In the cell of this embodiment, for example, assuming that the flow path 6 has a width of 0.1 mm and a depth of 0.05 mm, and the cell culture chamber 5 has a circular shape with a diameter of 1 mm and a depth of 0.05 mm, Has a considerably large capacity with respect to the flow path 6, so that the flow of the solution is very slow in this portion. For this reason, it is possible to trap cells at this location. In addition, the vicinity of the cell submerged at the bottom of the cell culture chamber 5 becomes a very quiet environment, and the cell easily adheres to the wall surface. If the cell culture chamber 5 has such a size, the cell culture chamber 5 is sufficiently small as compared with the conventional system, and a micro space characteristic of a microchip can be maintained. As in this embodiment, by forming the cell culture chamber 5 having a sufficiently large capacity for the flow channel 6 and not destroying the characteristics of the micro space, the cells are trapped in the micro space and proliferated. A microchip that can be operated can be realized.
[0030]
If the adhesion of the cells in the cell culture chamber 5 is insufficient even under these conditions, the adhesion can be promoted by modifying the inner surface of the flow path 6 and the inner surface of the cell culture chamber 5 using a chemical. is there.
[0031]
Further, before the substrates 1 and 2 are joined, chemical modification of the inner surface of only the cell culture chamber 5 can be performed to adjust conditions under which cells can easily adhere. Examples of such chemical modifications are collagen coat, poly-L-lysine coat, gelatin coat and the like. When such a chemical modification is performed on the bottom surface of the cell culture chamber 5, the cells can easily adhere to the cell culture chamber 5, which is convenient for culture.
[0032]
The manufacturing method of this embodiment will be briefly described below. The cells of the other embodiments can be manufactured by the same method.
An example of a method for manufacturing the cell of the present invention will be briefly described using a case where a glass substrate is used as an example.
[0033]
In order to form a groove or a concave portion for the flow path 6 and the cell culture chamber 5 in the glass substrates 1 and 2, an etching protection film is formed on the cleaned glass substrates 1 and 2 using a thin film forming apparatus (for example, a sputtering film forming apparatus). For example, a silicon (Si) thin film is formed, and a photoresist layer for patterning the etching protection film is formed thereon.
[0034]
Next, the photoresist is exposed using a photomask, and subsequently developed to pattern the photoresist. The exposure of the photoresist can be performed using an aligner generally used for manufacturing a semiconductor device.
[0035]
Next, the etching protection film is patterned using the photoresist pattern as a mask. If the etching protection film is silicon, SF ₆ Dry etching using high-frequency plasma in a gas can be used.
[0036]
Next, using the buttered etching protection film and photoresist as a mask, the glass substrates 1 and 2 are etched to form grooves and recesses. For etching the glass substrates 1 and 2, for example, a 46% hydrofluoric acid aqueous solution can be used as an etching solution.
Thereafter, the photoresist is removed, and the etching protection film is removed by etching.
[0037]
The processing for forming the through holes 3 and 4 in the glass substrate 1 can be performed by, for example, sandblasting.
The glass substrates 1 and 2 which have been subjected to the predetermined processing and thus prepared are overlapped and joined. As an example of the joining method, for example, a method in which a 1% hydrofluoric acid aqueous solution is interposed at the interface of the glass substrate and left at room temperature for about 24 hours while applying a load of about 1 MPa as necessary can be cited.
[0038]
FIG. 2 shows a second embodiment composed of two substrates. (A1) is a plan view of a surface outside the upper substrate 1a, (A2) is a plan view of a surface inside the same substrate 1a, (B1) is a plan view of a surface inside the lower substrate 2a, (B2) is a plan view of a surface outside the substrate 2a, (C1) is a plan view of a completed cell, (C2) is a cross-sectional view of the same cell taken along line XX, and (C3) is a view of the same cell. It is sectional drawing in the YY line position.
[0039]
In this embodiment, two substrates 1a and 2a are joined similarly to the embodiment of FIG. In this embodiment, only holes 3 and 4 for sample introduction and discharge are formed in the glass substrate 1a. On the other hand, the glass substrate 2a has a cell culture chamber 5a having a channel groove 6 having a width and a depth of several hundreds μm or less, and a circular recess having a diameter of about 1 mm, which is about the same as or deeper than the channel groove 6. Are formed on the same surface.
[0040]
Then, the surface on the inside of the substrate 1a and the surface of the substrate 2a where the flow channel 6 and the cell culture chamber 5a are formed face each other and are brought into close contact with each other, and are joined in the same manner as in the first embodiment. A chip type cell having a channel groove 6 for a sample and a cell culture chamber 5a is formed.
[0041]
FIG. 3 shows a third embodiment composed of two substrates.
It is obtained by joining two substrates 1b and 2b. In this embodiment, holes 3 and 4 for sample introduction and discharge are formed as through holes in the glass substrate 1b, and a flow channel 6 having a width and a depth of several hundred μm or less, A cell culture chamber 5b is formed on the same surface as a circular recess having a diameter of about 1 mm, which is about the same or deeper. On the other hand, the glass substrate 2b is a flat substrate. The flat surface of the substrate 2b is brought into close contact with the surface of the substrate 1a where the flow channel 6 and the cell culture chamber 5a are formed, and is brought into close contact with the substrate 2b. A chip type cell having a channel 6 and a cell culture chamber 5b is formed.
In this embodiment, the cell culture chamber 5b is formed above the flow channel 6.
[0042]
Depending on the type of cells, a relatively deep cell culture chamber may be required. In such a case, as shown in the fourth embodiment in FIG. 4, the depth of the cell culture chamber can be arbitrarily set using three substrates. 4, (A1) is a plan view of a surface outside the uppermost substrate 1, (A2) is a plan view of a surface inside the same substrate 1, and (B) is a plan view of a substrate 10 interposed therebetween. , (C) is a plan view of the lowermost substrate 12, (D1) is a plan view of the completed cell, (D2) is a cross-sectional view of the same cell taken along the line XX, and (D3) is a Y-line of the same cell. It is sectional drawing in the Y-line position.
[0043]
In the first glass substrate 1, which is the uppermost layer, a flow channel 6 and through holes 3 and 4 at both ends thereof are formed as in the embodiment of FIG. A through-hole serving as the cell culture chamber 7 is formed in the second glass substrate 10 sandwiched therebetween. The third glass substrate 12 serving as the lowermost layer is a glass substrate having a flat inner bonding surface.
[0044]
There is a cell culture chamber 7 in which three substrates 1, 10, and 12 are brought into close contact with each other such that the flow channel 6 of the substrate 1 is on the inside, and the depth is defined by the thickness of the second substrate 10. A cell is formed.
[0045]
Further, in the cell of the embodiment of FIG. 4, after forming a complicated structure in which components such as microvalves are formed using the first and second substrates 1 and 10, chemical modification is performed only at a necessary place. The third substrate 12 can be joined, and a cell having a complicated structure and chemically modified only at necessary places can be easily obtained.
[0046]
FIG. 5 shows a fifth embodiment including three substrates.
(A1) is a plan view of a surface outside the uppermost substrate 1a, (A2) is a plan view of a surface inside the same substrate 1, (B) is a plan view of a substrate 10a interposed therebetween, (C) Is a plan view of the lowermost substrate 12, (D1) is a plan view of a completed cell, (D2) is a cross-sectional view of the same cell at the XX line position, and (D3) is a cross-sectional view of the same cell at the YY line position. FIG.
[0047]
Through holes 3 and 4 serving as liquid inlets and outlets are formed in the first substrate 1a serving as the uppermost layer. In the second substrate 10a to be sandwiched, a through hole serving as a cell culture chamber 7 is formed between the channel groove 6 and both ends thereof. The third substrate 12 serving as the lowermost layer is a substrate having a flat inner bonding surface. The three substrates 1a, 10a, and 12 are bonded together so that the flow channel 6 of the substrate 10a faces the substrate 1a, and a cell having a cell culture chamber 7 is formed.
[0048]
In this embodiment, the flow channel 6 and the cell culture chamber 7 are formed on the second substrate 10a. As the second substrate 10a, for example, a silicon substrate that is easily microprocessed by lithography and etching is used. Processing of a silicon substrate is established by semiconductor manufacturing technology, and at present, fine processing of a submicron (1 μm or less) region can be easily performed. Since processing such as formation of a flow path is not required for the first substrate 1a, valves and the like can be designed regardless of the design of the flow path and the cell culture chamber. For example, PDMS (polydimethylsiloxane) is used as the first substrate 1a. Further, since the third substrate 12 is only required to be a flat substrate, there is a wide range of choices in both material and thickness. For example, Pyrex (registered trademark) glass having a thickness of about 0.15 mm Use those.
[0049]
A cell having such a combination of materials is suitable for microscopic observation. Further, Pyrex (registered trademark) glass has a property that cells are more easily adhered than silicon, and the flow path is silicon, and the bottom of the cell culture chamber is Pyrex (registered trademark) glass, which is suitable for cell culture.
[0050]
Further, the first substrate 1a can be bonded later, so that the second substrate 10a and the third substrate 12 are first bonded to make it easier for the cells to adhere to the bottom surface of the cell culture chamber. After the chemical modification is performed by dropping a collagen solution or the like on the first substrate, by joining the first substrate 1a to the second substrate 10a, there is also an advantage that the chemical modification becomes easy.
[0051]
FIG. 6 shows a sixth embodiment including three substrates. In this embodiment, a second substrate 10b having a large overall thickness is used, and only the portion where the cell culture chamber is formed is reduced in thickness, thereby increasing the overall mechanical strength. .
[0052]
FIG. 7 shows a seventh embodiment including three substrates.
In the second substrate 10c, a through hole serving as a cell culture chamber 7 is formed between the flow channel 6 and both ends thereof, and through holes 3 and 4 serving as liquid ports are also formed. The first substrate 1c serving as the uppermost layer and the third substrate 12b serving as the lowermost layer are substrates having a flat bonding surface. The three substrates 1c, 10c, and 12b are adhered to each other so that the flow channel 6 of the substrate 10c faces the substrate 12b and the first substrate 1c forms the upper surface of the cell culture chamber 7, and the cell culture is performed. A cell having a chamber 7 is formed.
[0053]
FIG. 8 shows an eighth embodiment including three substrates.
In the second substrate 10d, a through hole serving as a cell culture chamber 7 and through holes 3 and 4 serving as liquid inlets and outlets are formed, and a flow channel 6 is formed in a third substrate 12c serving as a lowermost layer. . The first substrate 1c to be the uppermost layer is a substrate having a flat bonding surface. The three substrates 1c, 10d, and 12c are bonded together so that the flow channel 6 of the substrate 12c faces the substrate 10d and forms the upper surface of the cell culture chamber 7 with the first substrate 1c. A cell having a chamber 7 is formed.
[0054]
In the embodiment shown in FIGS. 7 and 8, the cell culture chamber 7 is formed at a position higher than the flow path 6. In addition, a material more suitable for cell culture can be obtained by selecting a material to which cells easily adhere as the substrate 1c or by chemically modifying the inner surface of the cell culture chamber 7 of the substrate 1c.
[0055]
【The invention's effect】
According to the cell of the present invention, it is possible to trap a cell at a specific place with a simple structure. In addition, a function of culturing or activating cells can be provided only in this place. With this configuration, it is possible to provide a chip for easily testing the reaction of a target cell.
Since the cell of the present invention has a very simple structure and can be introduced into various manufacturing steps, it can be applied to a complicated microchip incorporating a valve or the like.
By using the cell of the present invention, there are many advantages such as a small sample size, a high-speed measurement, and simultaneous measurement of multiple samples.
[Brief description of the drawings]
FIG. 1 is a view showing a cell of a first embodiment, (A1) is a plan view of a surface outside an upper substrate, (A2) is a plan view of a surface inside the same substrate, and (B1). Is a plan view of a surface inside the lower substrate, (B2) is a plan view of a surface outside the same substrate, (C1) is a plan view of a completed cell, and (C2) is an XX line of the cell. (C3) is a cross-sectional view of the same cell taken along line YY.
FIG. 2 is a diagram showing a cell of a second embodiment, (A1) is a plan view of a surface outside the upper substrate, (A2) is a plan view of a surface inside the same substrate, and (B1). Is a plan view of a surface inside the lower substrate, (B2) is a plan view of a surface outside the same substrate, (C1) is a plan view of a completed cell, and (C2) is an XX line of the cell. (C3) is a cross-sectional view of the same cell taken along line YY.
FIG. 3 is a sectional view showing a cell according to a third embodiment.
4A and 4B are diagrams showing a cell according to a fourth embodiment, in which (A1) is a plan view of a surface outside the uppermost substrate, (A2) is a plan view of a surface inside the same substrate, and (B). ) Is a plan view of the substrate interposed therebetween, (C) is a plan view of the lowermost substrate, (D1) is a plan view of the completed cell, (D2) is a cross-sectional view of the same cell taken along line XX, (D3) is a sectional view of the same cell taken along line YY.
5A and 5B are diagrams showing a cell according to a fifth embodiment, in which (A1) is a plan view of a surface outside the uppermost substrate, (A2) is a plan view of a surface inside the same substrate, and (B). ) Is a plan view of the substrate interposed therebetween, (C) is a plan view of the lowermost substrate, (D1) is a plan view of the completed cell, (D2) is a cross-sectional view of the same cell taken along line XX, (D3) is a sectional view of the same cell taken along line YY.
FIG. 6 is a sectional view showing a cell according to a sixth embodiment.
FIG. 7 is a sectional view showing a cell according to a seventh embodiment.
FIG. 8 is a sectional view showing a cell according to an eighth embodiment.
[Explanation of symbols]
1, 1a, 1c, 2, 2a, 10, 10a, 10b, 10c, 10d, 12, 12b, 12c Substrate
3, 4 Through hole for liquid inlet / outlet
5,7 Cell culture room
6 Channel groove

Claims (13)

A channel formed inside the substrate and capable of flowing a liquid,
A liquid port connected to both ends of the flow path,
A cell culture chamber, which is provided in the middle of the flow path inside the base, has a step with respect to the flow path, and has a cross-sectional area orthogonal to a flow direction of liquid larger than a cross-sectional area of the flow path; A cell characterized by being provided integrally with the cell. The cell according to claim 1, wherein the depth of the step in the cell culture chamber is larger than the depth of the flow path. The base is composed of two substrates, the flow path is formed as a groove having a bottom on the surface of the upper substrate, the cell culture chamber is formed as a recess on the surface of the lower substrate, and both substrates are formed. 3. The cell according to claim 1, wherein the flow channel and the cell culture chamber are joined to be inside, and the liquid outlet is provided in the upper substrate. 4. The base is composed of two substrates, the upper substrate is provided with the liquid inlet / outlet, the lower substrate is formed on the same surface as a groove having a bottom, and the cell culture chamber is provided with the flow channel. 3. The cell according to claim 1, wherein the cell is formed as a concave portion having a bottom deeper than a channel, and both substrates are joined so that the channel and the cell culture chamber are on the inside. 4. The base body is composed of two substrates, the upper substrate is formed on the same surface, the channel is formed as a groove having a bottom, and the cell culture chamber is formed as a recess having a bottom deeper than the flow channel, The lower substrate is a substrate having a flat surface, and the flow path, the cell culture chamber, and the two substrates are joined such that the flat surface of the lower substrate is inside, and the liquid inlet / outlet is connected to the upper substrate. The cell according to claim 1, wherein a cell is provided. The cell according to claim 3, 4 or 5, wherein the inner surface of the cell culture chamber is subjected to a chemical modification for enhancing cell adhesion. The base is composed of three substrates, the uppermost substrate has the channel formed as a groove having a bottom on its surface, the intermediate substrate has a cell culture chamber formed as a through hole, and the lowermost substrate has a flat surface. A substrate having a surface, and these three substrates are joined so that the intermediate substrate is interposed therebetween, and the flow path forming surface of the uppermost substrate and the flat surface of the lowermost substrate are inside, 3. The cell according to claim 1, wherein the liquid port is provided in the uppermost substrate. The base is composed of three substrates, the uppermost substrate is provided with the liquid inlet / outlet, the intermediate substrate is formed with a groove having a bottom on its surface, and the cell culture chamber is formed as a through hole. The lowermost substrate is a substrate having a flat surface, these three substrates sandwich the intermediate substrate, the flow path forming surface of the intermediate substrate faces the uppermost substrate, and the lowermost substrate The cell according to claim 1, wherein the cell is joined such that the flat surface is on the inside. The substrate is composed of three substrates, the uppermost substrate is a substrate having a flat surface, the intermediate substrate is formed as a groove having a bottom on the surface thereof, and the cell culture chamber is formed as a through hole. The lowermost substrate is also a substrate having a flat surface, these three substrates sandwich the intermediate substrate, the flow path forming surface of the intermediate substrate faces the flat surface of the lowermost substrate, 3. The cell according to claim 1, wherein the uppermost substrate is joined so that a flat surface of the uppermost substrate faces the inside to close a through hole of the cell culture chamber, and the liquid inlet / outlet is provided in the intermediate substrate. The substrate is composed of three substrates, the uppermost substrate is a substrate having a flat surface, the cell culture chamber is formed as a through hole in the intermediate substrate, and the flow channel is formed on the surface of the lowermost substrate. These three substrates sandwich the intermediate substrate, the flow path forming surface of the lowermost substrate is on the inner side, and the flat surface of the uppermost substrate is on the inner side, and the cell culture chamber is formed. 3. The cell according to claim 1, wherein the cell is joined so as to close the through hole, and the liquid inlet / outlet is provided in the intermediate substrate. 4. The surface opposite to the surface where the flow path is present is chemically modified to enhance cell adhesion on a surface to be an inner surface of the cell culture chamber among surfaces of the substrate bonded to the intermediate substrate. 11. The cell according to any one of 7 to 10. The substrate which is joined to the intermediate substrate on a surface opposite to a surface where the flow path exists and which constitutes the cell culture chamber is made of a material having a higher cell adhesive force than the intermediate substrate. The cell described in Crab. The thickness of a substrate which is joined to the intermediate substrate on a surface opposite to a surface where the flow path exists and which constitutes the cell culture chamber is set to a thickness suitable for optical microscope observation. 13. The cell according to any one of items 1 to 12.

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