Cell culture system, cell culture apparatus, and cell imaging system

Abstract

Provided are a cell culture system, a cell culture apparatus, and a cell imaging system. A cell culture system includes: a cell culture apparatus including: a placement table on which a culture container having a culture surface on which cell culture can be performed and a side surface forming a peripheral wall around the culture surface is placed, the side surface being optically transparent; a supply pipe that allows the liquid to be supplied into the culture vessel; and a discharge pipe that allows the liquid to be discharged from the culture container. An imaging unit that allows observation of the culture surface; and a light irradiation unit that emits illumination light toward the side surface, in which the supply piping and the discharge piping are arranged such that an opening of the supply piping and an opening of the discharge piping are located near a point on an inner surface of the culture vessel that defines a width of the culture surface in a direction orthogonal to an optical axis of the illumination light.

Description

Technical Field

[0001] The present disclosure relates to a cell culture system, a cell culture device and a cell imaging system. Background Art

[0002] In cell culture, a technology capable of automatically observing a large area of ​​a culture surface is important for realizing automatic cell culture. In addition, when a colony is formed as a cell cluster when observing the culture surface, the shape of the colony of cultured cells can be observed based on the scattering of the irradiated light. In Japanese Patent Publication No. 2016-077226 (Patent Document 1), a system capable of observing the shape of a colony in the following manner is proposed: by irradiating light through the side of a culture container and observing the scattered light from the colony with a camera installed below the culture container.

[0003] Automatic cell culture equipment needs to supply water to the culture medium and drain water from the culture medium. Therefore, piping such as a hose is installed around the culture container. However, the system disclosed in Japanese Patent Laid-Open No. 2016-077226 is not constructed in consideration of the above piping. Summary of the invention

[0004] The present disclosure is made in view of the above circumstances. More specifically, an object of the present disclosure is to provide a cell culture apparatus, a cell culture system, and a cell imaging system that can realize automatic cell culture using piping for supplying and discharging water into and from a culture container.

[0005] In order to solve the above problems, according to one aspect of the present disclosure, a cell culture system is provided, comprising:

[0006] A cell culture system, comprising:

[0007] A cell culture apparatus comprising:

[0008] a placement table configured to allow placement of a culture container having a culture surface capable of performing cell culture and a side surface forming a peripheral wall around the culture surface, the side surface being optically transparent;

[0009] a supply pipe configured to allow liquid to be supplied into the culture container; and

[0010] a discharge pipe configured to allow liquid to be discharged from the culture container;

[0011] an imaging unit configured to allow observation of the culture surface; and

[0012] a light irradiation portion configured to emit illumination light toward the side surface,

[0013] The supply pipe and the exhaust pipe are arranged so that the opening of the supply pipe and the opening of the exhaust pipe are located near a point on the inner surface of the culture container, and the inner surface defines the width of the culture surface in a direction orthogonal to the optical axis of the illumination light.

[0014] Further features of the present invention will become apparent from the following description of exemplary embodiments with reference to the attached drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1A : is a diagram for illustrating a schematic configuration of a cell culture system according to a first embodiment of the present disclosure.

[0016] Figure 1B It is used to show Figure 1A is a diagram of a schematic configuration of a cell culture system as viewed from above.

[0017] Figure 2A It is a diagram for illustrating the positional relationship between the light irradiation section and the culture container according to the first embodiment.

[0018] Figure 2B It is used to show the Figure 2A Schematic diagram of the structure of the culture container and piping shown in FIG.

[0019] Figure 2C It is used to show the Figure 2A ] is a diagram of a schematic configuration of a culture container and piping shown in FIG. 1 with respect to the culture container in different configurations.

[0020] Figure 3A It is a diagram for illustrating the positional relationship between the light irradiation section and the culture container according to the first embodiment.

[0021] Figure 3B It is used to show the Figure 3A Schematic diagram of the structure of the culture container and piping shown in FIG.

[0022] Figure 3C It is used to show the Figure 3A ] is a diagram of a schematic configuration of a culture container and piping shown in FIG. 1 with respect to the culture container in different configurations.

[0023] Figure 4A It is a diagram for illustrating a modification example of light shielding of the bottom of the container according to the first embodiment.

[0024] Figure 4B It is a diagram for illustrating another modified example of light shielding of the bottom of the container according to the first embodiment.

[0025] Figure 5 It is a diagram for illustrating the positional relationship among the light irradiation unit, the culture container, and the pipe according to the second embodiment.

[0026] Figure 6 It is a diagram for illustrating the positional relationship among the light irradiation unit, the culture container, and the pipe according to the third embodiment.

[0027] Fig. 7A is a diagram for illustrating a schematic configuration of a cell culture system according to a fourth embodiment.

[0028] Figure 7B It is used to show the illumination light Fig. 7A FIG. 4 is a diagram of a cell culture system being irradiated. DETAILED DESCRIPTION

[0029] Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. However, for example, the size, material, shape, and relative position of the elements described in the following embodiments may be freely selected in addition to the specific conditions described below, and may be changed according to the configuration or various conditions of the device to which the present disclosure is applied. In addition, elements having substantially the same or functionally similar configurations in this specification and the accompanying drawings are indicated by the same reference numerals throughout the accompanying drawings, and their repeated descriptions are omitted.

[0030] In cell culture, the ratio of cells in the culture surface (cell occupancy rate) is used as an indicator of the timing of passage. Therefore, it is desirable to photograph cells as phase objects distributed over a large area in a short period of time. In order to observe a large area in a short period of time, a low-magnification lens is generally used. However, the use of a low-magnification lens is not suitable for a phase contrast microscope using transmitted light. Therefore, when photographing cells over a large area using a low-magnification lens, it is preferred to irradiate light through the side of the culture container and photograph the scattered light from the cells from the bottom side.

[0031] Furthermore, when automatic cell culture is performed, as described above, piping for supplying, for example, water to and from the culture medium is often provided to each culture container (e.g., a culture dish). It is thought that when parallel light capable of illuminating a large area is irradiated to the culture medium through the side of the culture dish in order to image the cells, a portion of the irradiated light may reach the piping and be reflected by the piping.

[0032] Therefore, when light is irradiated through the side of the culture container in order to image the cells, it is considered that the light reflected from the piping (hereinafter referred to as "stray light") needs to be considered. In addition, when parallel light is to pass through the side of the culture container to illuminate a large area of ​​the culture container, it is necessary to consider not only the stray light from the piping as exemplified above, but also the stray light generated due to, for example, the shape of the culture container (for example, a protrusion arranged on the bottom surface). As described above, in a cell culture device including, for example, piping, it is considered that a more appropriate cell image can be obtained by reducing the stray light generated by the piping or reducing the area that the stray light can reach. In order to cope with the problems caused by the presence of the piping, a structure that can reduce the stray light or reduce the area that may be affected by the stray light is described in the following embodiment.

[0033] <First Embodiment>

[0034] Here, refer to Figures 1A to 4B A cell culture system according to a first embodiment of the present disclosure is described. Figure 1A is a diagram for illustrating a cell culture system 100 according to the present embodiment. Figure 1B yes Figure 1A A plan view of the structure shown in when viewed from above (schematic structure in plan view). For ease of description, the culture container is maintained in a horizontal plane, and the top side, bottom side, right side, left side and side are defined based on the horizontal plane and the light irradiation direction in the following description. However, these definitions are not intended to limit the content of the present disclosure. Depending on, for example, the shape of the culture container, the light irradiation direction and the situation in which the cell culture system is installed, the above directions can be appropriately changed. In addition, the directions used for description are not completely accurate, but may include manufacturing errors (such as errors occurring when the culture container is placed on a placement table and errors in parallel associated with the parallelism of the illumination light) and errors allowed when performing the present disclosure.

[0035] (Construction of cell culture system)

[0036] like Figure 1A As shown in the schematic diagram of FIG. 1 , the cell culture system 100 according to the present embodiment includes a placement table 2, a light source 3, a camera 5, and a pipe 6. A culture container 1 for cells to be cultured is placed on the placement table 2. The culture container 1 is a storage container for storing a culture medium for culturing cells. A culture surface 1a that allows cells to attach and proliferate is provided on the bottom of the container. In the culture container 1, the culture surface 1a may be the bottom surface of the container or a component that is provided on the bottom surface of the container and is different from the container.

[0037] The placement table 2 has a hole 2a corresponding to the imaging area. The hole 2a allows the imaging device 5 arranged below the placement table 2 to image the cells in the culture container 1 through the transparent bottom surface of the culture container 1. The light source 3 emits an illumination light 4 from one side of the culture container 1, and the illumination light 4 will illuminate the cells through the side of the culture container 1. The imaging device 5 is located below the culture container 1 and takes an image of scattered light from below to obtain the shape of the colony, and the scattered light is generated by the scattering of light irradiated to the cells attached to the culture surface 1a. The piping 6 includes a supply piping 6a and a discharge piping 6b. The supply piping 6a allows water and samples (such as culture medium) to be supplied to the culture container 1. The discharge piping 6b allows water to be discharged from the culture container 1.

[0038] (Cultivation Container)

[0039] As described above, the culture container 1 stores the culture medium for culturing cells and the cells contained in the culture medium. Transparent resins and glasses that are easy to process and allow easy observation are suitable as materials for the culture container 1. When using resin, polystyrene, polycarbonate or acrylic acid can be used. In addition, as the shape of the culture container 1, a culture container having a rectangular shape or a circular shape on the bottom surface is exemplified. The culture surface 1a is formed to allow the attachment and proliferation of cells. A transparent material that allows easy observation is preferably used as the material of the culture surface 1a. In terms of cost and production, it is preferred that a part of the culture surface 1a is made of the same material as the culture container 1. In this case, it is preferred to perform surface treatment such as plasma treatment on the culture surface 1a so that the cells are more likely to attach only to the culture surface 1a.

[0040] (Light irradiation part)

[0041] Preferably, the light source 3 is capable of emitting light having a wavelength that allows the imaging device 5 to acquire images without adversely affecting the cells as the illumination light 4. A halogen lamp or an LED light source can be used as the light source 3. The light source 3 according to the present embodiment is exemplified as an example of a light irradiation portion in the present disclosure, which includes, for example, a collimation unit as described below, depending on the conditions. In view of the contrast of image quality, it is preferred to reduce stray light as much as possible to acquire only scattered light from the cells. Therefore, it is preferred that the illumination light 4 irradiates only the cells on the culture surface 1a. In addition, it is preferred that the imaging surface is free of unevenness. Therefore, it is preferred that the illumination light 4 has a substantially uniform intensity and wavelength range in the imaging area. As described above, it is preferred to select a light source that irradiates only a uniform amount of light to the culture surface. For example, it is preferred to use a linear LED light source with high directivity or a linear light guide including a bundle of optical fibers with a linear configuration at the output end.

[0042] Use of the above-mentioned light source 3 enables efficient illumination of only the culture surface 1a and its vicinity. The illumination light 4 emitted from the light source 3 irradiates the side of the culture container 1. Therefore, the illumination light 4 has a difference in illumination intensity between the incident side and the exit side of the culture container 1a. In order to reduce the difference, it is preferred to use an illumination light with a small expansion angle that is closer to parallel light. Therefore, it is effective to use a collimating unit for the light source 3 to obtain parallel light as the illumination light 4. For example, a cylindrical lens is used as a collimating unit. In addition, providing a plurality of light sources to realize multi-directional illumination is also effective for obtaining more uniform illumination light.

[0043] Furthermore, as a method of limiting the illumination area generated by the light source 3 to the culture surface 1a, it is also effective to provide a mask on the side of the culture container 1. The mask can be provided between the light source 3 and the culture surface 1a. When the mask is provided, it is not necessary to use the linear light emitted from the light source 3 as the illumination light 4. Therefore, when the mask is used, although the efficiency is reduced, a light source for emitting light capable of illuminating a large area can be selected.

[0044] (Camera equipment)

[0045] The imaging device 5 is used to acquire an image of cells attached to the culture surface 1 a. For example, a so-called digital camera using CMOS and CCD can be used as the imaging device 5 .

[0046] (Piping)

[0047] The piping 6 is used to supply a test solution for a culture medium into the culture container 1 or to discharge it from the culture container 1. In the present embodiment, a pair of piping 6, i.e., a supply piping 6a for supplying a test solution or water and a discharge piping 6b for discharging the test solution or water, is provided for one culture container 1. However, the number of piping is not limited to two, and a greater number of pipings may be provided for one culture container. When replacing the culture medium, the piping 6 may be used, for example, to supply water to the culture medium and to discharge water from the culture medium. When the illumination light 4 is incident on the piping 6, the light reflected from the piping 6 becomes stray light. The stray light may be a factor that reduces the quality of the cell image taken by the imaging device 5. Therefore, preferably, a material with low reflectivity is used for the piping 6 to reduce the above-mentioned stray light. It is preferred to use a transparent resin (e.g., silicone or polystyrene).

[0048] However, even when a material with the lowest possible reflectivity is selected, the intensity of the reflected light is often higher than the intensity of the scattered light from the cells. Therefore, it is preferred that the position of the pipe relative to the illumination light 4 be determined so that the illumination light with high intensity is not easily incident on the pipe 6 or the reflected light from the pipe 6 is not easily affected by the imaging area. The configuration of the pipe 6 in view of the above problem is described below with reference to Figures 2A to 3C describe.

[0049] FIG. 2A to FIG. 2C1 is a diagram for illustrating the positional relationship between the light source 3 and the culture container 1 and the positional relationship between the culture container 1 and the pipe 6 supplied to the culture container 1. Figure 2A Schematically shows the positional relationship between the light source 3 and the culture container 1 having a rectangular bottom surface (having a rectangular parallelepiped box shape). Figure 2B Schematically shows an example of the positional relationship between the side of the culture container 1 and the pipe 6. Figure 2C Another example of the positional relationship between the side surface of the culture container 1 and the pipe 6 is shown in FIG.

[0050] like Figure 2B As shown, the two pipes 6a and 6b are not arranged near the side 1b (which are the sides for the incident side and the exit side of the illumination light 4), but are respectively arranged near the side 1c other than the side 1b and parallel to the optical axis of the illumination light 4. The above arrangement of the pipes 6a and 6b allows the area that may be affected by stray light to be positioned outside the imaging area and as far away from the imaging area as possible, and thus a more appropriate cell image of a larger area can be obtained.

[0051] In order to reduce the influence of stray light as much as possible, it is desirable that the pipe 6 is arranged in contact with the side surfaces 1c that are parallel to each other. However, considering the process of bringing the pipe 6 closer to the culture container 1, it is actually impractical to arrange the pipe 6 in contact with the culture container 1. Therefore, the pipe 6 is arranged near the side surfaces 1c that are parallel to each other. Here, the term "near" defines a state in which the pipe 6 is arranged near the side surface 1c without causing a problem when performing the above process.

[0052] Next, refer to Figure 2C A specific configuration of the piping 6 is described, which can reduce the influence of stray light to a level at which the colony can be observed. Here, the width of the culture surface 1a of the culture container 1 in the direction perpendicular to the optical axis is represented by D, and the straight lines extending in the extension direction of the inner surface of the culture container 1 parallel to the optical axis are represented by L1 and L2, respectively. At this time, the piping 6 is arranged at a position close to the inner surface of the culture container 1 that defines the width D. More specifically, the piping 6 is arranged so that in a plan view, the distance between the foot of the perpendicular line extending from the center of the opening of the piping 6a to the culture surface 1a and the one of the straight lines L1 and L2 that is closer to the piping 6a is equal to D / 10 or less, and similarly, the distance between the foot of the perpendicular line extending from the center of the opening of the piping 6b to the culture surface 1a and the one of the straight lines L1 and L2 that is closer to the piping 6b is equal to D / 10 or less. When the piping 6 is arranged to satisfy the above conditions, the piping 6 can be easily brought close to the culture container 1, and a suitable cell image of a large area can be obtained.

[0053] Next, refer to FIG. 3A to FIG. 3CThe arrangement of the pipe 6 is described when the culture container 1 has a circular bottom surface (a cylindrical shape with a circular cross section). Figure 3A FIG. 2 schematically shows the positional relationship between the light source 3 and the culture container 1 in the above case. Figure 3B and Figure 3C Schematically showing the positional relationship between the side surface of the culture container 1 and the pipe 6.

[0054] When the culture container 1 has a circular bottom surface, the diameter of the culture surface 1a in the direction perpendicular to the optical axis is represented by D, and the straight lines parallel to the optical axis extending in the extending direction of the tangent line of the inner surface of the culture container 1 are represented by L1 and L2, respectively. At this time, the pipe 6 is arranged at a position near the inner surface of the culture container 1 defining the width D. More specifically, the pipe 6a is arranged so that the distance between the foot of the perpendicular line extending from the center of the opening of the pipe 6a to the culture surface 1a and the one of the straight lines L1 and L2 closer to the pipe 6a in a plan view is equal to D / 10 or less, and similarly, the pipe 6b is arranged so that the distance between the foot of the perpendicular line extending from the center of the opening of the pipe 6b to the culture surface 1a and the one of the straight lines L1 and L2 closer to the pipe 6b in a plan view is equal to D / 10 or less. When the pipe 6 is arranged to satisfy the above conditions, even in the culture container 1 having a circular bottom surface, the pipe 6 can be easily brought close to the culture container 1, and a suitable cell image of a large area can be obtained.

[0055] Furthermore, when the piping 6 is arranged as follows, the influence of stray light can be reduced to a level at which the colony can be observed. More specifically, when the culture container 1 has a circular bottom surface, the area between the circle and the side of the square inscribed in the circle extending perpendicular to the optical axis in a plan view is defined as an area 1d where the piping 6 is not allowed to be arranged. In this case, when the piping 6 is arranged between the side extending parallel to the optical axis and the inner surface of the culture container 1, the area that may be affected by stray light can be positioned outside the imaging area and as far away from the imaging area as possible. Therefore, a more appropriate cell image of a larger area can be obtained.

[0056] In the first embodiment, a cell culture system 100 has been described in which cell culture is performed while the light source 3, the placement table 2, the imaging device 5, and the piping 6 are fixed. However, the configuration applicable to the present disclosure is not limited to the above-mentioned cell culture system. For example, a placement table 2 and a piping 6 corresponding to one culture container 1 are provided to the culture container 1. The piping 6 and the culture container 1 are configured to satisfy the above-mentioned positional relationship, and a cell culture device having a plurality of the above-mentioned configurations can also be formed. An imaging system including a light source 3 and an imaging device 5 corresponding thereto can be added to the above-mentioned cell culture device. The light source 3 is configured to emit illumination light 4 to satisfy the above-mentioned relationship relative to the piping 6. In this case, it is desirable that the imaging system and the cell culture device can change their positions relative to each other so that the imaging system can capture images of cells in, for example, a plurality of culture containers. When the above-mentioned imaging system is additionally provided to the cell culture device, a cell imaging system according to the present disclosure can be constructed.

[0057] As described above, the cell culture system 100 according to the present disclosure includes a cell culture device, a camera unit (camera 5) and a light irradiation unit (light source 3). In this case, the cell culture device includes a placement table 2 and a piping 6, and the piping 6 includes a supply piping 6a and a discharge piping 6b. The culture container 1 is placed on the placement table 2. The culture container 1 has a culture surface 1a on which cell culture can be carried out and a side surface forming a peripheral wall around the culture surface 1a. The side surface is optically transparent. The supply piping 6a is used when liquid is supplied to the culture container, and the discharge piping 6b is used when liquid is discharged from the culture container. The camera 5 is used to observe the culture surface 1a. The illumination light 4 irradiated toward the side of the culture container 1 is emitted from the light source 3. In the cell culture system 100 according to the first embodiment, the supply piping 6a and the exhaust piping 6b are configured so that the opening of the supply piping 6a and the opening of the exhaust piping 6b are located near a point on the inner surface of the culture container 1, which defines the width D of the culture surface 1a in a direction orthogonal to the optical axis of the illumination light 4.

[0058] When an image of cells is to be obtained, the following conditions are preferably satisfied as conditions for reducing the influence of stray light from the piping on the image to an appropriate level. The width of the culture surface 1a is represented by D, and two straight lines passing through a point on the inner surface of the culture container 1 and parallel to the optical axis are represented by L1 and L2. In this case, it is preferred that the supply piping 6a and the exhaust piping 6b are arranged so that the distance between the foot of a perpendicular line extending from the center of the opening of one of the pipings 6a and 6b to the culture surface 1a and the one of the straight lines L1 and L2 that is closer to the corresponding piping is equal to D / 10 or less, and the distance between the foot of a perpendicular line extending from the center of the opening of the other of the pipings 6a, 6b to the culture surface 1a and the one of the straight lines L1 and L2 that is closer to the corresponding piping is equal to D / 10 or less.

[0059] In addition, in the cell culture system 100, the culture container 1 may have a rectangular shape (rectangular bottom surface, i.e., rectangular culture surface) in a plan view. In this case, the supply piping 6a and the exhaust piping 6b are preferably arranged near the inner side of the side 1c of the culture container 1 extending parallel to the optical axis. In addition, the culture container 1 may have a circular shape (circular bottom surface, i.e., circular culture surface) in a plan view. In this case, the supply piping 6a and the exhaust piping 6b are preferably arranged between the two opposite surfaces of the square frame inscribed in the circle parallel to the optical axis and the inner surface of the circle opposite to the two opposite surfaces in a plan view. The culture container 1 having a rectangular or circular shape includes two areas that meet the above conditions. When at least one of the supply piping 6a or the exhaust piping 6b is arranged in any one of these areas, the other piping may be arranged in another area of ​​these areas. More specifically, both the supply piping 6a and the exhaust piping 6b may be arranged in one of the two areas that meet the above conditions. Alternatively, one of the supply piping 6 a and the discharge piping 6 b may be arranged in any one of the above-mentioned areas, and the other of the supply piping 6 a and the discharge piping 6 b may be arranged in the other of the above-mentioned areas.

[0060] In the above-mentioned cell culture system 100, it is desirable to illuminate only the cells with the illumination light 4, for example, to prevent stray light from being generated due to irradiating an area other than the desired area of ​​the culture container 1. Therefore, the illumination light 4 is preferably a sheet of parallel light. In addition, the present disclosure can also be applied to a cell culture device including the above-mentioned placement table 2, the supply piping 6a and the exhaust piping 6b. In this case, the present disclosure can be applied to a cell imaging system including a cell culture device, a light irradiation unit (light source 3) and an imaging unit (imaging device 5). In this case, it is preferred that the supply piping 6a and the exhaust piping 6b are configured so that their openings are located near a point on the inner surface of the culture container 1, which defines the width of the culture surface 1a in a direction orthogonal to the central axis of the culture container 1 placed on the placement table 2 in a plan view. The light irradiation unit only needs to move relative to the cell culture device and can illuminate the side of the culture container 1 with illumination light in a manner that matches the optical axis with the above-mentioned central axis. The imaging device 5 only needs to be able to capture an image in response to the illumination light to observe the culture surface irradiated by the illumination light.

[0061] <Modification of the first embodiment>

[0062] For example, it is conceivable to use a commercially available culture dish as the culture container 1. Commercially available culture dishes generally have protrusions on the circumference of their lower side to prevent the bottom surface of the culture dish from contacting the ground. When illumination light is incident on the protrusions, the light propagates in multiple directions, which may generate stray light. The purpose of the modification is to cope with the problem that occurs when a culture dish having the above-mentioned protrusions is used. Figure 4A and Figure 4B Configurations included in modified examples are described. Figure 4A and Figure 4B Corresponding to the first embodiment Figure 1A Therefore, the same constructions are denoted by the same reference numerals, and descriptions thereof are omitted here.

[0063] exist Figure 4A In the first modification shown, the culture container 1 has a protrusion 1e on its bottom surface. When the illumination light 4 emitted from the light source 3 is incident on the protrusion 1e, the above-mentioned stray light is generated. In this modification, a mask 8 is provided between the protrusion 1e and the light source 3 to shield the protrusion 1e from being irradiated by the illumination light 4. In this way, the protrusion 1e is not irradiated by the illumination light 4, so that the stray light generated by the protrusion 1e can be prevented from being captured as an image by the camera system 5.

[0064] In addition, the method of shielding the protrusion 1e from being irradiated with the illumination light is not limited to using the mask 8. For example, a structure may be added in which the protrusion 1e is set outside the irradiation area with the illumination light on the placement stage 2. Figure 4B This configuration is described as a second modification. Figure 4B In the structure shown, the placement table 2 has a recess 2b, the depth of which corresponds to the thickness of the protrusion 1e (the distance from the bottom to the top of the protrusion in a direction perpendicular to the bottom surface of the culture container 1). When the culture container 1 is placed on the above-mentioned placement table 2, the protrusion 1e is not illuminated by the illumination light, and only the culture surface 1a is illuminated by the illumination light emitted from one side. An example is given of a recess formed to receive the entire circumferential area of ​​the bottom of the culture container 1. However, the recess can be formed to receive only the protrusion 1e. The shape of the recess is not limited to the example described here, as long as it can avoid being illuminated by the illumination light.

[0065] When the cell culture apparatus according to the first embodiment is provided with a configuration in which the protrusions formed on the bottom surface of the culture container 1 are shielded from illumination light as described above, stray light due to the protrusions can be reduced. Therefore, a more appropriate cell image of a larger area can be obtained.

[0066] As described above, the cell culture system 100 according to the present disclosure includes a cell culture device, a camera unit (camera 5) and a light irradiation unit (light source 3). In the cell culture system, the cell culture device includes a placement table 2 and a piping 6, and the piping 6 includes a supply piping 6a and a discharge piping 6b. The culture container 1 is placed on the placement table 2. The culture container 1 has a culture surface 1a on which cell culture can be performed and a side surface forming a peripheral wall near the culture surface 1a. The side surface is optically transparent. The supply piping 6a is used when liquid is supplied to the culture container, and the discharge piping 6b is used when the liquid is discharged from the culture container. The camera 5 is used to observe the culture surface 1a. The illumination light 4 irradiated toward the side of the culture container 1 is emitted from the light source 3. In the cell culture system according to the modified example, the culture container 1 has a protrusion 1e, which is formed on the surface opposite to the culture surface 1a and protrudes in the direction opposite to the direction toward the culture surface 1a. The placement table 2 may have a structure that shields the protrusion 1e from the illumination light 4. As the configuration, for example, a mask 8 or a recess 2b can be used. The mask 8 is provided between the light source 3 and the protrusion 1e. The recess 2b is formed in the surface of the placement table 2 and receives the protrusion 1e when the culture container 1 is placed on the placement table 2.

[0067] <Second Embodiment>

[0068] In the first embodiment described above, for example, the supply pipe 6a used when supplying water to the culture container and the discharge pipe 6b used when discharging water from the culture container are arranged to be separated from each other. Meanwhile, in this embodiment, the supply pipe 6a and the discharge pipe 6b are arranged together as a set. Figure 5 The cell culture system 200 having the above-mentioned configuration according to the present embodiment is described. Figure 5 In Figure 1B The main structure is shown in the same way as in FIG.

[0069] like Figure 5 As shown, in the present embodiment, the supply piping 6a and the exhaust piping 6b are arranged to be in contact with each other. The supply piping 6a and the exhaust piping 6b are brought close to the culture container 1 as a group of piping 6. In the present embodiment, when the conditions for the configuration of one of the piping 6a and 6b described in the first embodiment are applied to the piping 6 as a group of piping 6 provided integrally, the same effect as that obtained in the first embodiment is obtained. When two pipings are provided integrally as a group, the number of sources of stray light can be reduced to one. Therefore, compared with the first embodiment, the effect of further reducing the influence of stray light can be obtained. Figure 5 In the embodiment, a culture container 1 having a rectangular bottom surface is shown by way of example. FIG. 3A to FIG. 3C The described configuration or condition may also be applicable to Figures 3A to 3CThe case of a culture container 1 with a circular bottom is shown by way of example in FIG.

[0070] <Third Embodiment>

[0071] The first and second embodiments have described configurations for reducing stray light when taking images of cultured cells, which are achieved by configuring the piping 6 with respect to the culture container 1 so that water is supplied or discharged, for example, through an opening located above the culture container 1. In the above configuration, a commercially available culture container can generally be used as the culture container 1. At the same time, it is also conceivable to process the culture container to reduce stray light. In the present embodiment, two holes (openings) for supplying water and discharging water are formed in the side of the culture container, and the piping is connected to the openings, respectively. Specifically, the piping 6 is directly connected to the opening formed in the side of the culture container. Refer to Figure 6 The cell culture system 300 having the above-mentioned configuration according to the present embodiment is described. Figure 6 In Figure 1B The main structure is shown in the same way as in FIG.

[0072] In this embodiment, the culture container 301 has a hole 301f corresponding to the supply pipe 6a and a hole 301g corresponding to the discharge pipe 6b. Figure 6 When the culture container 301 exemplarily shown in FIG. 1 has a rectangular bottom surface, holes 301f and 301g are formed in the side surface extending in the direction parallel to the optical axis of the illumination light 4. The arrangement of the holes and the connection of the pipes through the holes can substantially prevent the pipes 6 from appearing in the irradiation area of ​​the illumination light 4. Specifically, in the present embodiment, when an image of the cultured cells is captured, the pipes 6 are prevented from being arranged in the irradiation area of ​​the illumination light 4. Therefore, stray light generated by the illumination light 4 being incident on the pipes 6 can be reduced.

[0073] Reference Figure 6 A culture container 301 having a rectangular bottom surface is described. When the culture container 301 has a circular bottom surface, preferably, for example, the culture container 301 defines Figure 3BHoles 301a and 301b are formed on the inner surface of the culture container 301 with a width D in the direction of the width D. Alternatively, the holes 301a and 301b are preferably formed in the inner surface of the culture container 301 in a manner that falls within a range of D / 10 from the inner surface of the culture container 301 that defines the width D. As a result, stray light generated by the illumination light 4 being incident on the piping 6 can be reduced. The connection of both the supply piping 6a and the exhaust piping 6b through the hole formed in the side of the culture container 301 has been described. However, the present embodiment is not limited to the illustrated configuration. One of the supply piping 6a and the exhaust piping 6b can be arranged above the culture container 301 as in the first embodiment and other embodiments. Alternatively, the holes 301a and 301b can be formed in the side of the culture container 301 in a side-by-side arrangement.

[0074] <Fourth Embodiment>

[0075] In the first and second embodiments, when photographing cultured cells, the pipes 6 provided to the culture container 1 are not moved from their positions during cell culture, and the configuration for coping with stray light generated in this case has been described. Meanwhile, in the present embodiment, by withdrawing the pipes 6 from the irradiation area of ​​the illumination light 4 when photographing cultured cells, the possibility of generating stray light is reduced. Here, Fig. 7A and Figure 7B China and Israel Figure 1A The configuration of the cell culture system 400 according to the present embodiment is shown in the same manner as in FIG.

[0076] exist Fig. 7A , a cell culture system 400 is shown during cell culture. The light source 3 does not emit illumination light, and the pipe 6 is arranged at a position that allows water to be supplied to and discharged from the cultured cells. Figure 7B , a cell culture system 400 is shown when photographing cells. Illumination light 4 is emitted from a light source 3, and a pipe 6 is withdrawn upward from the irradiation area by a pipe drive unit (not shown). The above configuration prevents stray light from being generated due to the presence of the pipe 6, and a suitable cell image of a larger area can be obtained.

[0077] As described above, in the cell culture system 400 according to the present embodiment, when the culture surface 1a is observed by the imaging device 5, the supply pipe 6a and the exhaust pipe 6b are driven to move away from the culture container 1. As a result, the opening of the supply pipe 6a and the opening of the exhaust pipe 6b are located outside the irradiation area of ​​the illumination light, and the pipes themselves are also located outside the irradiation area. Therefore, stray light due to the presence of the pipe 6 is not generated, and a suitable cell image of a large area can be obtained.

[0078] The present disclosure is described above with reference to the embodiments. However, the present disclosure is not limited to the above embodiments. The present disclosure also includes inventions modified within the scope of the present disclosure, and inventions equivalent to the present disclosure. In addition, within the scope of the gist of the present invention, the above embodiments can be appropriately combined with each other.

[0079] According to one aspect of the present disclosure, a cell culture apparatus, a cell culture system, and a cell imaging system capable of realizing automatic cell culture using piping for supplying and draining water into and from a culture container can be provided.

[0080] While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments.The scope of the following claims is to be accorded the broadest interpretation so as to encompass all such modifications and equivalent structures and functions.

Claims

1. A cell culture system, comprising: A cell culture apparatus comprising: a placement table configured to allow placement of a culture container having a culture surface capable of performing cell culture and a side surface forming a peripheral wall around the culture surface, the side surface being optically transparent; a supply pipe configured to allow liquid to be supplied into the culture container; and a discharge pipe configured to allow liquid to be discharged from the culture container; an imaging unit configured to allow observation of the culture surface; and a light irradiation portion configured to emit illumination light toward the side surface, The supply pipe and the exhaust pipe are arranged so that the opening of the supply pipe and the opening of the exhaust pipe are located near a point on the inner surface of the culture container, and the inner surface defines the width of the culture surface in a direction orthogonal to the optical axis of the illumination light.

2. The cell culture system according to claim 1, in, When the width of the culture surface is represented by D, and two straight lines passing through the point on the inner side of the culture container and parallel to the optical axis are represented by L1 and L2, the supply piping and the exhaust piping are configured so that, in a plan view, the distance between the foot of a perpendicular line extending from the center of the opening of the supply piping to the culture surface and one of the straight lines L1 and L2 that is closer to the supply piping, and the distance between the foot of a perpendicular line extending from the center of the opening of the exhaust piping to the culture surface and one of the straight lines L1 and L2 that is closer to the exhaust piping are both equal to D / 10 or less.

3. The cell culture system according to claim 1, wherein: When the culture container has a rectangular shape in a plan view, the supply pipe and the exhaust pipe are arranged near the inner side of a side surface of the culture container extending parallel to the optical axis.

4. The cell culture system according to claim 1, wherein: When the culture container has a rectangular shape in a plan view, the supply pipe and the exhaust pipe are connected to the culture container in a manner having openings exposed on a side surface of the culture container extending parallel to the optical axis.

5. The cell culture system according to claim 1, wherein: When the culture container has a circular shape in plan view, the supply pipe and the exhaust pipe are both arranged between one of two opposing surfaces parallel to the optical axis of a frame having a square shape in plan view inscribed in the circle and a portion of a surface opposite to the one opposing surface defining the inside of the circle, or One of the supply piping and the exhaust piping is arranged between one of the two relative surfaces parallel to the optical axis and a portion of the surface opposite to the one relative surface that defines the inner side of the circle, and the other of the supply piping and the exhaust piping is arranged between the other of the two relative surfaces and a portion of the surface opposite to the other relative surface that defines the inner side of the circle.

6. The cell culture system according to claim 1, wherein: When the culture container has a circular shape in plan view, the supply pipe and the exhaust pipe are connected to the culture container so that the supply pipe and the exhaust pipe each have an opening exposed on a portion of a surface defining an inner side of the circle, the portion being opposed to one of two opposing surfaces of a frame having a square shape in plan view inscribed in the circle and parallel to the optical axis, or One of the supply piping and the exhaust piping has an opening exposed on a portion of the surface defining the inner side of the circle, the portion being opposite to one of the two relative surfaces parallel to the optical axis, and the other of the supply piping and the exhaust piping has an opening exposed on a portion of the surface defining the inner side of the circle, the portion being opposite to the other of the two relative surfaces.

7. A cell culture system, comprising: A cell culture apparatus comprising: a placement table that allows placement of a culture container, the culture container having a culture surface capable of performing cell culture and a side surface forming a peripheral wall around the culture surface, the side surface being optically transparent; a supply pipe configured to allow liquid to be supplied into the culture container; and a discharge pipe configured to allow liquid to be discharged from the culture container; an imaging unit configured to allow observation of the culture surface; and a light irradiation portion configured to emit illumination light toward the side surface, When the culture surface is observed by the imaging unit, the opening of the supply pipe and the opening of the discharge pipe are located outside an irradiation area of ​​the illumination light. 8 . The cell culture system according to claim 7 , wherein the opening of the supply pipe and the opening of the exhaust pipe are exposed on the side surface of the culture container.

9. The cell culture system according to claim 7, wherein: When the culture surface is observed by the imaging unit, the supply pipe and the discharge pipe are driven to move away from the culture container so that the opening of the supply pipe and the opening of the discharge pipe are located outside the irradiation area of ​​the illumination light.

10. The cell culture system according to claim 1, wherein the illumination light is a sheet of parallel light.

11. The cell culture system according to claim 1, wherein the culture container has a protrusion formed on a surface opposite to the culture surface and protruding in a direction opposite to a direction toward the culture surface, and The cell culture system further comprises a structure for shielding the protrusion from the illumination light.

12. A cell culture device comprising: a placement table that allows placement of a culture container, the culture container having a culture surface capable of performing cell culture and a side surface forming a peripheral wall around the culture surface, the side surface being optically transparent; a supply pipe configured to allow liquid to be supplied into the culture container; and a discharge pipe configured to allow liquid to be discharged from the culture container, The supply piping and the exhaust piping are configured so that the opening of the supply piping and the opening of the exhaust piping are located near points on the inner surface of the culture container, and the inner surface defines the width of the culture surface in a direction orthogonal to the central axis of the culture container placed on the placement table in a plan view.

13. A cell imaging system, comprising: The cell culture device according to claim 12; a light irradiation section configured to move relative to the cell culture apparatus and capable of emitting the illumination light toward the side surface in such a manner that an optical axis of the illumination light matches the central axis; and An imaging section is configured to allow observation of the culture surface illuminated by the illumination light in response to the illumination light.

Images