Cell culture system, cell culture device, and cell imaging system

Abstract

To provide a cell culture system corresponding to automated cell culture using piping for water supply and drainage to a culture vessel.SOLUTION: A cell culture device having a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side surface constituting a peripheral wall of the culture surface is mounted, supply piping for supplying liquid into the culture vessel, and discharge piping for discharging liquid from the culture vessel; an imaging unit for observing the culture surface; and a light irradiation unit for irradiating the side surface with illumination light are arranged in a cell culture system, and openings of the supply piping and the discharge piping are arranged close to a point inside the culture vessel that defines a width of the culture surface in a direction perpendicular to an optical axis of the illumination light.SELECTED DRAWING: Figure 2

Description

[Technical field]

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

[0002] When culturing cells, a technology for automatically observing a wide area of ​​the culture surface is an important technology for realizing automatic cell culturing. In addition, when a colony of cells is formed, it is also possible to observe the shape of the colony of cultured cells from the scattering of irradiated light. Patent Document 1 proposes a system that can observe the colony shape by irradiating light from the side of the culture vessel and observing the scattered light from the colony with a camera installed at the bottom of the culture vessel. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 20016-077226 A Summary of the Invention [Problem to be solved by the invention]

[0004] Here, the automatic cell culture device requires supplying and discharging the culture medium, and therefore piping such as tubes is installed around the culture vessel. However, the system disclosed in Patent Document 1 is not configured in consideration of such piping.

[0005] The present disclosure has been made in consideration of the above-mentioned circumstances. Specifically, one of the objectives of the present disclosure is to provide a cell culture device, a cell culture system, and a cell imaging system that are compatible with automatic cell culture using piping for supplying and discharging water to and from a culture container. [Means for solving the problem]

[0006] In order to solve the above problems, a cell culture system according to one embodiment of the present disclosure includes: a cell culture device including: a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side constituting a peripheral wall of the culture surface is mounted; a supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging the liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, The openings of the supply pipe and the discharge pipe are arranged close to a point on the inside of the culture vessel that defines the width of the culture surface in a direction perpendicular to the optical axis of the illumination light. Effect of the Invention

[0007] According to one aspect of the present disclosure, it is possible to provide a cell culture device, a cell culture system, and a cell imaging system that are compatible with automatic cell culture using piping for supplying and discharging water to a culture container. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 is a diagram showing a schematic configuration of a cell culture system according to a first embodiment of the present disclosure. [Diagram 2] FIG. 2 is a diagram showing the positional relationship between a light irradiation unit, a culture vessel, and piping in the first embodiment. [Diagram 3] FIG. 2 is a diagram showing the positional relationship between a light irradiation unit, a culture vessel, and piping in the first embodiment. [Figure 4] 4A to 4C are diagrams illustrating a light-shielding aspect of the container bottom in the first embodiment. [Diagram 5] FIG. 13 is a diagram showing the positional relationship between a light irradiation unit, a culture vessel, and piping in the second embodiment. [Figure 6] FIG. 13 is a diagram showing the positional relationship between a light irradiation unit, a culture vessel, and piping in the third embodiment. [Figure 7] FIG. 13 is a diagram showing a schematic configuration of a cell culture system according to a fourth embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. However, the dimensions, materials, shapes, and relative positions of components described in the following embodiments are arbitrary except for the specific conditions described below, and can be changed according to the configuration of the device to which the present disclosure is applied or various conditions. In addition, in this specification and drawings, components having substantially the same or functionally similar configurations are given the same reference numerals between the drawings to avoid redundant description.

[0010] In cell culture, the ratio of cells occupying the culture surface (cell occupancy rate) is an indicator of the timing of passage, etc., and it is desirable to image cells, which are phase objects distributed over a wide area, in a short time. Low-magnification lenses are usually used to observe a wide area in a short time, but the use of low-magnification lenses is not suitable for phase-contrast microscopes that use transmitted light. Therefore, when photographing cells over a wide area with a low-magnification lens, it is preferable to irradiate light from the side of the culture vessel and image the scattered light from the cells from the bottom side.

[0011] Furthermore, when performing automated cell culture, as described above, piping for supplying and discharging water and the like to the culture medium is often provided for each culture vessel (e.g., petri dish).When cells are imaged by irradiating the culture medium with parallel light with a wide irradiation area from the side of the petri dish, it is considered that part of the irradiated light reaches the piping and is reflected from the piping.

[0012] Therefore, when imaging cells by irradiating light from the side of the culture vessel, it is considered necessary to consider the light reflected from this piping (hereinafter referred to as stray light). In addition, when attempting to irradiate a wide area of ​​the culture vessel with parallel light from the side of the culture vessel, it is necessary to consider stray light caused by the shape of the culture vessel (e.g., protrusions arranged on the bottom surface) in addition to the exemplified piping. From the above, it is considered that in a cell culture device having piping, etc., a more suitable cell image can be obtained by reducing the stray light generated by the piping, etc., or by reducing the range of the stray light. In the embodiment described below, a configuration that can reduce such stray light or its range of influence will be described as a measure against this piping.

[0013] First Embodiment Hereinafter, a cell culture system according to a first embodiment of the present disclosure will be described with reference to FIGS. 1 to 4. FIG. 1(a) shows a schematic configuration of a cell culture system 100 according to this embodiment, and FIG. 1(b) shows a plan view (schematic configuration in plan view) of the configuration shown in FIG. 1(a) seen from above. In the following description, for convenience, the culture vessel is held in a horizontal plane, and the upper, lower, left, right, side, etc. are defined based on this horizontal plane and the direction of light irradiation, but these are not intended to limit the contents of the disclosure. These directions can be changed as appropriate depending on the shape of the culture vessel, the direction of light irradiation, and the installation situation of the cell culture system. In addition, the directions used in the description are not strict, and include manufacturing errors, for example errors that occur when the culture vessel is placed on the placement table, parallel errors related to the parallelism of the illumination light, etc., and each includes an error that is permissible when the disclosure is executed.

[0014] (Configuration of cell culture system) As shown in the schematic diagram of FIG. 1(a), a cell culture system 100 according to this embodiment includes a mounting table 2, a light source 3, an imaging device 5, and piping 6. A culture vessel 1 for culturing cells is mounted on the mounting table 2. The culture vessel 1 is a holding vessel for holding a medium for culturing cells, and a culture surface 1a is provided at the bottom of the vessel to which cells can adhere and grow. In the culture vessel 1, the culture surface 1a may be the bottom of the vessel, or may be a separate structure from the vessel disposed at the bottom of the vessel.

[0015] The mounting table 2 is provided with a hole 2a corresponding to the imaging area so that the imaging device 5 arranged below the mounting table 2 can image the cells in the culture vessel 1 through the bottom surface of the transparent culture vessel 1. The light source 3 emits illumination light 4 from the side direction of the culture vessel 1, which is irradiated onto the cells through the side surface. The imaging device 5 images the scattered light, which is irradiated onto the cells attached to the culture surface 1a, from below the culture vessel 1, and obtains the shape of the colony, etc. The piping 6 includes a supply piping 6a for supplying samples such as water and culture medium into the culture vessel 1, and a discharge piping 6b for draining the water.

[0016] (Culture container) As described above, the culture vessel 1 holds a medium for culturing cells and cells arranged in the medium. The material of the culture vessel 1 is preferably a transparent resin or glass that is easy to process and observe. Resins such as polystyrene, polycarbonate, and acrylic can be used. Examples of the shape include a rectangular or circular bottom surface. The culture surface 1a is configured so that cells can attach and grow, and is preferably made of a transparent material that is easy to observe. In terms of cost and ease of production, it is preferable to configure a part of the culture vessel 1 with the same material as the culture vessel 1. In this case, it is preferable to perform a surface treatment such as plasma treatment only on the culture surface 1a to make it easier for cells to attach.

[0017] (Light irradiation part) The light source 3 is preferably one that can capture an image with the imaging device 5 and can emit light of a wavelength that does not adversely affect cells as the illumination light 4, and a halogen lamp, an LED light source, or the like can be used. The light source 3 in this embodiment is exemplified as one aspect of the light irradiation unit in the present disclosure, including, for example, a collimating means described below, depending on the conditions. From the viewpoint of image quality contrast, it is preferable to reduce stray light as much as possible in order to capture only scattered light from the cells. Therefore, it is preferable that the illumination light 4 is irradiated only to the cells on the culture surface 1a. In addition, it is preferable that there is no unevenness in the captured image surface, and therefore, it is preferable that the illumination light 4 has an approximately uniform intensity and wavelength range in the imaging area. From the above, it is preferable to select a light source that irradiates a uniform amount of light only to the culture surface. For example, it is preferable to use a highly directional line-type LED light source or a line light guide in which the emission end of the bundle fiber is arranged in a line.

[0018] By using such a light source 3, it is possible to efficiently illuminate only the vicinity of the culture surface 1a. Since the illumination light 4 from the light source 3 is irradiated from the side of the culture vessel 1, a difference in illumination intensity occurs between the entrance side and exit side of the irradiated light on the culture surface 1a. In order to reduce this difference, it is preferable to use light that is closer to parallel and has a reduced spread angle of the illumination light. Therefore, it is effective to use a collimating means for the light source 3 to obtain parallel light as the illumination light 4. An example of the collimating means is a cylindrical lens. In addition, preparing multiple light sources and illuminating from multiple directions is also effective in uniforming the illumination light.

[0019] Furthermore, as a method for limiting the illumination range of the light source 3 to the culture surface 1a, it is also effective to place a mask on the side of the culture vessel 1, which can be placed between the light source 3 and the culture surface 1a. When a mask is placed, the need for the illumination light 4 from the light source 3 to be linear is reduced. Therefore, when a mask is used, it is possible to select a light source that emits light that can illuminate a wide range, although the efficiency is reduced.

[0020] (Imaging device) The imaging device 5 is used to obtain images of cells attached to the culture surface 1a. As the imaging device 5, a so-called digital camera using a CMOS, a CCD, or the like can be used.

[0021] (Plumbing) The piping 6 is used to supply or discharge a test solution such as a culture medium into the culture vessel 1. In this embodiment, two pairs of piping, a supply piping 6a used to supply a test solution, water, etc. and a discharge piping 6b used to discharge these, are arranged for one culture vessel 1. However, the number of piping is not limited to this, and more piping may be arranged for one culture vessel. The piping 6 can be used for suction and drainage of the culture medium during culture medium replacement, but when the illumination light 4 is irradiated onto the piping 6, the reflected light becomes stray light. This stray light can be a factor in reducing the quality of the image of the cells captured by the imaging device 5. Therefore, in order to reduce such stray light, it is preferable to use a material with low reflectance for the piping 6, and it is preferable to use a transparent resin (for example, silicone, polystyrene, etc.).

[0022] However, even if a material with the lowest possible reflectance is selected, the intensity of the reflected light is often higher than the scattered light from the cells. Therefore, it is preferable to devise a position of the piping relative to the illumination light 4 so that the piping 6 is unlikely to be irradiated with strong illumination light or the reflected light from the piping 6 is unlikely to affect the imaging range. The arrangement of the piping 6 taking such a point into consideration will be described below with reference to FIG. 2 or FIG. 3.

[0023] Fig. 2 shows the positional relationship between the culture vessel 1 and the piping 6 with respect to the piping 6 provided for the culture vessel. Fig. 2(a) shows the positional relationship between the light source 3 and the culture vessel 1 when the culture vessel 1 has a rectangular bottom (square box shape), and Fig. 2(b) shows a schematic example of the positional relationship between the side surface of the culture vessel 1 and the piping 6. Fig. 2(c) shows another example of the positional relationship between the side surface of the culture vessel 1 and the piping 6.

[0024] 2(b), the two pipes 6a, 6b are arranged not to be close to the side surface 1b on the entrance side or exit side of the illumination light 4, but to be close to the side surface 1c parallel to the optical axis of the illumination light 4 other than the side surface 1b. By arranging in this manner, it is possible to position the range affected by stray light as far outside the imaging range as possible, and it is possible to obtain a more suitable wide-range cell image.

[0025] In order to reduce the influence of stray light as much as possible, it is desirable to arrange these pipes 6 in contact with the parallel side surfaces 1c. However, when considering the process of accessing the pipes 6 to the culture vessel 1, it is not realistic to actually have them in contact. For this reason, the pipes 6 are arranged close to the parallel side surfaces 1c, but the proximity described here specifies a state in which the pipes 6 are arranged close to the side surfaces 1c to the extent that no problems arise in carrying out the subsequent process.

[0026] Next, a specific arrangement of the piping 6 that can reduce the influence of stray light to a level where colony observation can be performed will be described with reference to FIG. 2(c). Here, the width of the culture surface 1a in the culture vessel 1 in the direction perpendicular to the optical axis is defined as width D, and the straight lines located in the extension direction of the inner surface of the culture vessel 1 parallel to the optical axis are defined as lines L1 and L2. In this case, the piping 6 is arranged in a position close to the inner surface of the culture vessel 1 that defines the width D, more specifically, within a range where the distance between the foot of the perpendicular line drawn from the opening center of the piping 6a and 6b to the culture surface 1a and the closer of the lines L1 and L2 is D / 10 or less. By arranging the piping 6 so as to satisfy this condition, the access of the piping 6 to the culture vessel 1 is facilitated, and a suitable wide range of cell images can be obtained.

[0027] Next, the arrangement of the piping 6 when the culture vessel 1 has a circular bottom (circular cylinder) will be described with reference to Fig. 3. The positional relationship between the light source 3 and the culture vessel 1 in this case is shown in Fig. 3(a), and the positional relationship between the side surface of the culture vessel 1 and the piping 6 is shown in Fig. 3(b) or Fig. 3(c).

[0028] When the culture vessel 1 has a circular bottom surface, the width of the culture surface 1a in the direction perpendicular to the optical axis is defined as D, and the straight lines located in the extending direction of the tangent to the inner surface of the culture vessel 1 parallel to the optical axis are defined as L1 and L2. In this case, the pipe 6 is arranged in a position close to the inner surface of the culture vessel 1 that defines the width D, more specifically, within a range where the distance between the foot of the perpendicular line dropped from the opening center of the pipes 6a and 6b to the culture surface 1a and the closer of the straight lines L1 and L2 is D / 10 or less. By arranging the pipe 6 so as to satisfy this condition, even in the culture vessel 1 having a circular bottom surface, the pipe 6 can easily access the culture vessel 1, and a suitable wide range of cell images can be obtained.

[0029] Furthermore, even when the piping 6 is arranged as follows, the influence of stray light can be reduced to a level where colony observation can be performed. Specifically, when the culture vessel 1 has a circular bottom, the area outside the sides extending perpendicular to the optical axis of the square inscribed in the circle in a plan view is defined as an area 1d where the piping 6 cannot be arranged. In this case, by arranging the piping 6 inside the culture vessel 1, outside the sides extending parallel to the optical axis, it is possible to position the area affected by stray light as far outside the imaging range as possible. Therefore, it is possible to obtain a more suitable wide range of cell images.

[0030] In the first embodiment, the cell culture system 100 is described in which cell culture is performed with the light source 3, the mounting table 2, the imaging device 5, and the piping 6 fixed. However, the configuration to which the present disclosure is applied is not limited to such a cell culture system. For example, a cell culture device having a plurality of such configurations can be configured in which a corresponding mounting table 2 and piping 6 are arranged for one culture vessel 1, and the piping 6 and the culture vessel 1 are configured to satisfy the above-mentioned positional relationship. An imaging system consisting of a light source 3 that emits illumination light 4 to the piping 6 so as to satisfy the above-mentioned relationship, and a corresponding imaging device 5 can be added to such a cell culture device. In this case, it is desirable to change the relative positions of the imaging system and the cell culture device so that the imaging system can image cells and the like in a plurality of culture vessels. Then, by adding such an imaging system to the cell culture device, the present disclosure can also configure a cell imaging system.

[0031] As described above, the cell culture system 100 according to the present disclosure includes a cell culture device, an imaging unit (imaging device 5), and a light irradiation unit (light source 3). In this case, the cell culture device includes a mounting table 2 and piping 6 including a supply piping 6a and a discharge piping 6b. A culture vessel 1 having a culture surface 1a on which cells can be cultured and an optically transparent side surface constituting the peripheral wall of the culture surface 1a is mounted on the mounting table 2. The supply piping 6a is used when supplying liquid into the culture vessel, and the discharge piping 6b is used when discharging liquid from the culture vessel. The imaging device 5 is used when observing the culture surface 1a, and the light source 3 emits illumination light 4 that is irradiated onto the side surface of the culture vessel 1. In the cell culture system 100 in the first embodiment, the openings of the supply piping 6a and the discharge piping 6b are arranged so as to be close to a point inside the culture vessel 1 that defines the width D of the culture surface 1a in a direction perpendicular to the optical axis of the illumination light 4.

[0032] In addition, when obtaining an image of cells, it is preferable to satisfy the following conditions for reducing the influence of stray light from the piping on the image to a suitable level. The width of the culture surface 1a is D, and two straight lines passing through an inner point and parallel to the optical axis are line L1 and line L2. In this case, it is preferable to arrange the supply pipe 6a and the discharge pipe 6b so that the distance from the foot of a perpendicular line drawn from the center of their openings to the culture surface 1a to the closer of line L1 or line L2 is D / 10 or less.

[0033] Further, in the cell culture system 100, the culture vessel 1 may have a rectangular shape in plan view (the bottom surface, i.e., the culture surface, is rectangular). In such a case, the supply pipe 6a and the discharge pipe 6b may be arranged close to the inside of the side surface 1c of the culture vessel 1 arranged parallel to the optical axis. Also, the culture vessel 1 may have a circular shape in plan view (the bottom surface, i.e., the culture surface, is circular). In such a case, the supply pipe 6a and the discharge pipe 6b may be arranged between a parallel opposing surface in a square frame in plan view inscribed in a circle having an opposing surface parallel to the optical axis and a surface that constitutes the inside of the circle opposing the opposing surface. Furthermore, there are two ranges that satisfy this condition in a rectangular or circular culture vessel 1, and when at least one of the supply pipe 6a and the discharge pipe 6b is arranged in one of these ranges, the other pipe can be arranged in the other range. Specifically, both the supply pipe 6a and the discharge pipe 6b can be arranged together in the two ranges that satisfy the above condition. Alternatively, one of the supply pipe 6a and the discharge pipe 6b may be disposed in one of these ranges, and the other of the supply pipe 6a and the discharge pipe 6b may be disposed in the other of these ranges.

[0034] In the cell culture system 100 described above, the illumination light 4 is preferably irradiated only to the cells in order to avoid stray light caused by irradiation of areas other than the desired area of ​​the culture vessel 1, and may be a sheet-like parallel light. The present disclosure may also be applied to a cell culture device including the above-mentioned mounting base 2, the supply pipe 6a, and the discharge pipe 6b. In this case, the present disclosure may also be applied to the cell culture device as a cell imaging system by adding a light irradiation unit (light source 3) and an imaging unit (imaging device 5). In this case, the openings of the supply pipe 6a and the discharge pipe 6b may be disposed close to a point on the inside of the culture vessel 1 that defines the width of the culture surface 1a in a direction perpendicular to the central axis of the culture vessel 1 mounted on the mounting base 2 in a plan view. The light irradiation unit may move relative to the cell culture device and irradiate the illumination light to the side of the culture vessel 1 so that the optical axis coincides with the central axis. The imaging device 5 may capture an image for observing the culture surface illuminated by the illumination light in response to the illumination light.

[0035] <Modification of the first embodiment> As the culture vessel 1, for example, a commercially available petri dish or the like may be used. Many commercially available petri dishes have protrusions around the lower side to prevent the bottom of the dish from touching the ground, and when illumination light is irradiated onto these protrusions, the light spreads in many directions, which may cause stray light. This modified example aims to deal with the case where a petri dish having such protrusions is used. Note that, although FIG. 4(a) or 4(b) is referred to for the configuration included in this modified example, these figures correspond to FIG. 1(a) of the first embodiment, and the same reference numerals are used for the same configurations, and the description here is omitted.

[0036] In the first modification shown in FIG. 4(a), the culture vessel 1 has a protrusion 1e on its bottom surface. When the illumination light 4 emitted from the light source 3 is irradiated onto this protrusion 1e, the above-mentioned stray light occurs. In this modification, a mask 8 is disposed between the protrusion 1e and the light source 3 so as to block the illumination light 4 from being irradiated onto the protrusion 1e. This prevents the illumination light 4 from being irradiated onto the protrusion 1e, and prevents the stray light caused by the protrusion 1e from being captured by the imaging device 5.

[0037] In addition, the method of blocking the illumination light from being applied to the projection 1e is not limited to the specifications of the mask 8, and for example, a configuration may be added to the mounting base 2 to remove the projection 1e from the illumination light irradiation range. Such an embodiment will be described as Modification 2 with reference to FIG. 4(b). In the configuration shown in FIG. 4(b), the mounting base 2 is provided with a recess 2b having a depth corresponding to the thickness of the projection 1e (the distance from the bottom of the projection to the top in the direction perpendicular to the bottom surface of the culture vessel 1). When the culture vessel 1 is placed on such a mounting base 2, the illumination light is not applied to the projection 1e, and the illumination light is applied only to the culture surface 1a from the side. Here, an embodiment in which a recess corresponding to the entire bottom area of ​​the culture vessel 1 is provided is exemplified, but it is also possible to make the recess accommodate only the projection 1e, and the shape of the recess is not limited to the example shown here as long as it is possible to avoid irradiation with the illumination light.

[0038] By providing the cell culture device of the first embodiment with a configuration for blocking illumination light from reaching the protrusions provided on the bottom surface of the culture vessel 1 as described above, it is possible to reduce stray light caused by the protrusions, thereby making it possible to obtain a more suitable wide-area cell image.

[0039] As described above, the cell culture system 100 according to the present disclosure includes a cell culture device, an imaging unit (imaging device 5), and a light irradiation unit (light source 3). In this case, the cell culture device includes a mounting base 2 and piping 6 including a supply piping 6a and a discharge piping 6b. A culture vessel 1 having a culture surface 1a on which cells can be cultured and an optically transparent side surface constituting a peripheral wall of the culture surface 1a is mounted on the mounting base 2. The supply piping 6a is used when supplying liquid into the culture vessel, and the discharge piping 6b is used when discharging liquid from the culture vessel. The imaging device 5 is used for observing the culture surface 1a, and the light source 3 emits illumination light 4 that is irradiated onto the side surface of the culture vessel 1. In the cell culture system according to this modification, the culture vessel 1 has a protrusion 1e provided on the opposite surface of the culture surface 1a and protruding in the opposite direction to the culture surface 1a. The mounting base 2 can have a configuration that blocks the illumination light 4 from the protrusion 1e. In this configuration, for example, a mask 8 disposed between the light source 3 and the projection 1e, and a recess 2b provided on the surface of the mounting table 2 for accommodating the projection 1e when the culture vessel 1 is mounted thereon can be used.

[0040] <Second embodiment> In the above-mentioned first embodiment, the supply pipe 6a used for supplying water or the like to the culture vessel and the discharge pipe 6b used for discharging water are arranged separately. In contrast, in this embodiment, the supply pipe 6a and the discharge pipe 6b are arranged together. The cell culture system 200 according to this embodiment having such a configuration will be described with reference to FIG. 5, which shows the main configuration in the same format as FIG. 1(b).

[0041] As shown in FIG. 5, in this embodiment, the supply pipe 6a and the discharge pipe 6b are provided in contact with each other, and are configured to access the culture vessel 1 as a pipe 6 in an integrated manner. In this embodiment, the same effect as in the first embodiment can be obtained by applying the conditions for the arrangement of one of the pipes 6a and 6b described in the first embodiment to the integrated pipe 6. In addition, by integrating the two pipes, the source of stray light is reduced to one, and the effect of further reducing the influence of stray light compared to the first embodiment can be obtained. Note that, although FIG. 5 illustrates an example in which the culture vessel 1 has a rectangular bottom surface, the configuration or conditions described with reference to FIG. 3 can also be applied to the case in which the culture vessel 1 has a circular bottom surface as illustrated in FIG. 3.

[0042] <Third embodiment> In the first and second embodiments, a configuration was described in which stray light is reduced during imaging of cultured cells in a configuration in which a pipe 6 is arranged to supply or discharge water or the like from an opening above the culture vessel 1. In these configurations, a culture vessel 1 that is normally commercially available can be used. In contrast, it is also possible to process the culture vessel to make it possible to reduce stray light. In this embodiment, two holes (openings) for supplying and discharging water or the like are provided on the side of the culture vessel, and pipes are connected to each of these. In other words, the pipe 6 is directly connected to the opening on the side of the culture vessel. The cell culture system 300 according to this embodiment having such a configuration will be described with reference to FIG. 6, which shows the main configuration in the same format as FIG. 1(b).

[0043] In this embodiment, the culture vessel 301 has a hole 301f corresponding to the supply pipe 6a and a hole 301g corresponding to the discharge pipe 6b. When the culture vessel 301 illustrated in FIG. 6 has a rectangular bottom surface, these holes 301f and 301g are provided on the side surface arranged parallel to the optical axis direction of the illumination light 4. By providing holes in such an arrangement and connecting the pipes through the holes, it is possible to almost completely eliminate the presence of the pipes 6 within the irradiation range of the illumination light 4. That is, in this embodiment, the pipes 6 are not arranged in the irradiation area of ​​the illumination light 4 when imaging the cultured cells. This makes it possible to reduce stray light caused by the illumination light 4 being irradiated onto the pipes 6.

[0044] In FIG. 6, the case where the culture vessel 301 has a rectangular bottom surface is described. In the case where the culture vessel 301 has a circular bottom surface, for example, holes 301a and 301b may be provided on the inner surface of the culture vessel 301 that defines the width D in FIG. 3(b). Alternatively, the holes 301a and 301b may be provided on the inner surface of the culture vessel 301 in a range of D / 10 in the direction of the width D from the inner surface of the culture vessel 301 that defines the width D. This makes it possible to reduce stray light caused by the illumination light 4 being irradiated onto the piping 6. In addition, here, the case where both the supply piping 6a and the discharge piping 6b are connected from holes on the side surface of the culture vessel 301 is described. However, this embodiment is not limited to the exemplified form, and one of them may be arranged above the culture vessel 1 as in the first embodiment. Alternatively, the holes 301a and 301b may be provided on the side surface of the culture vessel 301 so as to be aligned side by side.

[0045] <Fourth embodiment> In the first and second embodiments, a pipe 6 provided for the culture vessel 1 does not move from the position during cell culture even when the cultured cells are photographed, and a countermeasure is described for the case where stray light occurs as a result. In contrast, in this embodiment, the pipe 6 is moved away from the irradiation area of ​​the illumination light 4 when the cultured cells are photographed, thereby reducing the possibility of stray light occurring. Hereinafter, the configuration of a cell culture system 400 according to this embodiment is shown in FIG. 7 in a format similar to that of FIG. 1(a).

[0046] Fig. 7(a) shows the cell culture system 400 during cell culture. The light source 3 does not emit illumination light, and the piping 6 is placed in a position where water and the like can be supplied to and discharged from the cultured cells. Fig. 7(b) shows the cell culture system 400 during cell photography. Illumination light 4 is emitted from the light source 3, and the piping 6 is retracted upward from the irradiation area by a piping drive unit (not shown). With this configuration, stray light caused by the presence of the piping 6 is eliminated, making it possible to obtain a suitable wide-range cell image.

[0047] As described above, in the cell culture system 400 according to this embodiment, when the culture surface 1a is observed by the imaging device 5, the supply pipe 6a and the discharge pipe 6b are driven to move away from the culture container 1. As a result, the openings of the supply pipe 6a and the discharge pipe 6b are positioned outside the irradiation range of the illumination light, and these pipes themselves are also positioned outside the irradiation range. This prevents stray light caused by the presence of the pipe 6, making it possible to obtain a suitable wide-range cell image.

[0048] The above disclosure includes the following configurations. (Configuration 1) a cell culture device including: a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side constituting a peripheral wall of the culture surface is mounted; a supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging the liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, A cell culture system, wherein openings of the supply pipe and the discharge pipe are positioned adjacent to a point inside the culture vessel that defines the width of the culture surface in a direction perpendicular to the optical axis of the illumination light. (Configuration 2) When the width of the culture surface is D, and two straight lines passing through the inner point and parallel to the optical axis are line L1 and line L2, The cell culture system of configuration 1, wherein the supply pipe and the discharge pipe are arranged such that the distance from the foot of a perpendicular line drawn from the center of the opening of the supply pipe and the discharge pipe to the culture surface, respectively, to the closer of the straight line L1 and the straight line L2, is D / 10 or less. (Configuration 3) The cell culture system of configuration 1 or 2, wherein when the culture vessel has a rectangular shape in a planar view, the supply pipe and the discharge pipe are arranged adjacent to the inside of a side surface arranged parallel to the optical axis. (Configuration 4) A cell culture system as described in configuration 1, wherein when the culture vessel has a rectangular shape in a planar view, the supply pipe and the discharge pipe are connected to the culture vessel so that each opens on the inside of a side surface arranged parallel to the optical axis. (Configuration 5) The cell culture system of configuration 1 or 2, wherein, when the culture vessel has a circular shape in a planar view, the supply pipe and the discharge pipe are both arranged between one of the parallel opposing surfaces in a square frame in a planar view that has opposing surfaces parallel to the optical axis and is inscribed in the circle, and a surface that constitutes the inside of the circle that faces the one of the opposing surfaces; or, when one of the supply pipe and the discharge pipe is arranged between one of the parallel opposing surfaces and a surface that constitutes the inside of the circle that faces the one of the opposing surfaces, the other is arranged between the other of the opposing surfaces and a surface that constitutes the inside of the circle that faces the other of the opposing surfaces. (Configuration 6) The cell culture system of configuration 1, wherein when the culture vessel has a circular shape in a planar view, the supply pipe and the discharge pipe are connected to the culture vessel such that, when the culture vessel has opposing surfaces parallel to the optical axis and is inscribed in the circle, both of them open into a surface that constitutes the inside of the circle facing one of the parallel opposing surfaces in a square frame in a planar view inscribed in the circle, or when one of the supply pipe and the discharge pipe opens into a surface that constitutes the inside of the circle facing one of the parallel opposing surfaces, the other opens into a surface that constitutes the inside of the circle facing the other of the opposing surfaces. (Configuration 7) a cell culture device including: a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side constituting a peripheral wall of the culture surface is mounted; a supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging the liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, A cell culture system, wherein when the culture surface is observed by the imaging unit, an opening of the supply pipe and an opening of the discharge pipe are located outside the irradiation range of the illumination light. (Configuration 8) 8. The cell culture system according to claim 7, wherein the opening of the supply pipe and the opening of the discharge pipe are provided on a side surface of the culture vessel. (Configuration 9) The cell culture system of configuration 7, wherein when the culture surface is observed by the imaging unit, the supply pipe and the discharge pipe are driven away from the culture vessel, so that the opening of the supply pipe and the opening of the discharge pipe are positioned outside the irradiation range of the illumination light. (Configuration 10) 10. The cell culture system according to any one of configurations 1 to 9, wherein the illumination light is a sheet-shaped parallel light. (Configuration 11) the culture vessel has a protrusion provided on a surface opposite to the culture surface and protruding in a direction opposite to the culture surface, 11. The cell culture system according to any one of configurations 1 to 10, further comprising a configuration for blocking the illumination light from the protrusion. (Configuration 12) A mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side surface constituting a peripheral wall of the culture surface is mounted; A supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging liquid from the culture vessel, A cell culture device, wherein openings of the supply pipe and the discharge pipe are positioned close to a point inside the culture vessel that defines the width of the culture surface in a direction perpendicular to the central axis of the culture vessel placed on the mounting table in a planar view. (Configuration 13) A cell culture device according to configuration 12, A cell imaging system comprising: a light irradiation unit that moves relative to the cell culture device and can irradiate illumination light onto the side surface so that the optical axis coincides with the central axis; and an imaging unit that observes the culture surface illuminated by the illumination light in response to the illumination light. (Configuration 14) a cell culture device including: a culture vessel having a culture surface on which cells can be cultured, the side surface constituting the peripheral wall of the culture surface being optically transparent, the culture vessel being placed on a mounting base having a protrusion provided on the opposite side of the culture surface and protruding in the opposite direction from the culture surface; a supply pipe for supplying liquid into the culture vessel; and a discharge pipe for discharging liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, The cell culture system, wherein the mounting table has a configuration that blocks the illumination light from reaching the protrusion.

[0049] Although the present disclosure has been described above with reference to the embodiments, the present disclosure is not limited to the above-mentioned embodiments. Inventions modified within the scope of the present disclosure and inventions equivalent to the present disclosure are also included in the present disclosure. In addition, the above-mentioned embodiments can be appropriately combined within the scope of the present invention. [Explanation of symbols]

[0050] 1: culture vessel, 2: placement stand, 3: light source, 4: illumination light, 5: imaging device, 6: piping, 8: mask, 100, 200, 300, 400: cell culture system

Claims

1. a cell culture device including: a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side constituting a peripheral wall of the culture surface is mounted; a supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging the liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, A cell culture system, wherein openings of the supply pipe and the discharge pipe are positioned adjacent to a point inside the culture vessel that defines the width of the culture surface in a direction perpendicular to the optical axis of the illumination light.

2. When the width of the culture surface is D, and two straight lines passing through the inner point and parallel to the optical axis are a straight line L1 and a straight line L2, The cell culture system according to claim 1, wherein the supply pipe and the discharge pipe are arranged so that the distance from the foot of a perpendicular line drawn from the center of the opening of the supply pipe and the discharge pipe to the culture surface to the closer of the line L1 and the line L2, respectively, is D / 10 or less.

3. The cell culture system according to claim 1 , wherein when the culture vessel has a rectangular shape in a planar view, the supply pipe and the discharge pipe are arranged adjacent to an inner side of a side surface arranged parallel to the optical axis.

4. The cell culture system of claim 1 , wherein when the culture vessel has a rectangular shape in a planar view, the supply pipe and the discharge pipe are connected to the culture vessel so that each opens on a side that is arranged parallel to the optical axis.

5. 2. The cell culture system of claim 1, wherein when the culture vessel has a circular shape in a planar view, the supply pipe and the discharge pipe are both arranged between one of the parallel opposing surfaces in a square frame in a planar view that has opposing surfaces parallel to the optical axis and is inscribed in the circle, and a surface that constitutes the inside of the circle that faces the one of the opposing surfaces; or when one of the supply pipe and the discharge pipe is arranged between one of the parallel opposing surfaces and a surface that constitutes the inside of the circle that faces the one of the opposing surfaces, the other is arranged between the other of the opposing surfaces and a surface that constitutes the inside of the circle that faces the other of the opposing surfaces.

6. 2. The cell culture system of claim 1, wherein when the culture vessel has a circular shape in a planar view, the supply pipe and the discharge pipe are connected to the culture vessel such that, when the culture vessel has opposing surfaces parallel to the optical axis and is inscribed in the circle, both of the supply pipe and the discharge pipe open into a surface that constitutes the inside of the circle facing one of the parallel opposing surfaces in a square frame in a planar view that has opposing surfaces parallel to the optical axis and is inscribed in the circle, or when one of the supply pipe and the discharge pipe opens into a surface that constitutes the inside of the circle facing one of the parallel opposing surfaces, the other opens into a surface that constitutes the inside of the circle facing the other of the opposing surfaces.

7. a cell culture device including: a mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side constituting a peripheral wall of the culture surface is mounted; a supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging the liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, A cell culture system, wherein when the culture surface is observed by the imaging unit, an opening of the supply pipe and an opening of the discharge pipe are located outside an irradiation range 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 discharge pipe are provided on a side surface of the culture vessel.

9. The cell culture system of claim 7, wherein when the culture surface is observed by the imaging unit, the supply pipe and the discharge pipe are driven away from the culture vessel, so that the opening of the supply pipe and the opening of the discharge pipe are positioned outside the irradiation range of the illumination light.

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

11. the culture vessel has a protrusion provided on a surface opposite to the culture surface and protruding in a direction opposite to the culture surface, The cell culture system according to claim 1 , further comprising a configuration for blocking the illumination light from the protrusion.

12. A mounting base on which a culture vessel having a culture surface on which cells can be cultured and an optically transparent side surface constituting a peripheral wall of the culture surface is mounted; A supply pipe for supplying a liquid into the culture vessel; and a discharge pipe for discharging liquid from the culture vessel, A cell culture device, wherein openings of the supply pipe and the discharge pipe are positioned close to a point inside the culture vessel that defines the width of the culture surface in a direction perpendicular to the central axis of the culture vessel placed on the mounting table in a planar view.

13. The cell culture device according to claim 12 , A cell imaging system comprising: a light irradiation unit that moves relative to the cell culture device and can irradiate illumination light onto the side surface so that the optical axis coincides with the central axis; and an imaging unit that observes the culture surface illuminated by the illumination light in response to the illumination light.

14. a cell culture device including: a culture vessel having a culture surface on which cells can be cultured, the side surface constituting the peripheral wall of the culture surface being optically transparent, the culture vessel being placed on a mounting base having a protrusion provided on the opposite side of the culture surface and protruding in the opposite direction from the culture surface; a supply pipe for supplying liquid into the culture vessel; and a discharge pipe for discharging liquid from the culture vessel; An imaging unit for observing the culture surface; a light irradiation unit that irradiates the side surface with illumination light, The cell culture system, wherein the mounting table has a configuration that blocks the illumination light from reaching the protrusion.

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