JP7415371B2 - Cell culture imaging device and cell culture imaging method - Google Patents

Description

The present invention relates to a cell culture imaging device and a cell culture imaging method, and particularly to a cell culture imaging device and a cell culture imaging method for imaging cells cultured in a well.

Conventionally, cell culture devices for culturing cells in wells have been known (for example, see Non-Patent Document 1).

The above-mentioned Non-Patent Document 1 discloses a microfluidic chip (cell culture device) that includes a substrate layer in which a plurality of wells (microwells) are formed and an uppermost layer provided so as to cover the substrate layer. In this microfluidic chip, the substrate layer and the top layer are spaced apart from each other and are arranged along the horizontal direction. A gap between the substrate layer and the top layer constitutes a channel through which the cell suspension flows. Further, in the substrate layer, the microwell is formed to be depressed vertically downward. Then, some of the cells contained in the cell suspension flowing through the channel sink into one microwell. The remaining cells that do not settle into one microwell settle into other downstream microwells.

Furthermore, in the above-mentioned Non-Patent Document 1, the substrate layer and the uppermost layer are made of PDMS (Polydimethylsiloxane). PDMS is transparent and has the property of substantially not interacting with living organisms (cells).

Although not specified in Non-Patent Document 1, generally when observing (imaging) cells cultured in microwells, images are taken using a microscope from vertically below the microwells, for example.

Yu-Chih Chen, Xia Lou, Zhixiong Zhang, Patrick Ingram & Euisik Yoon “High-Throughput Cancer Cell Sphere Formation for Char “acterizing the Efficacy of Photo Dynamic Therapy in 3D Cell Cultures” Scientific Reports, 2015, 5, 12175.

When images of cells cultured in conventional microwells as described in Non-Patent Document 1 are imaged with a microscope from vertically below the microwells, cells (cell spheroids) placed below the microwells are observed. ) can be imaged. In other words, it is possible to image the horizontal growth of cells using a microscope. However, when cells are imaged from below, there is a problem in that it is not possible to image how cells grow in the direction along the direction of gravity (vertical direction).

This invention was made to solve the above-mentioned problems, and one purpose of this invention is to provide a cell system that allows imaging of how cells grow in the vertical direction. An object of the present invention is to provide a culture imaging device and a cell culture imaging method.

In order to achieve the above object, a cell culture imaging device according to a first aspect of the present invention includes a plate-shaped cell culture section having a main surface provided with a plurality of concave wells in which cells are cultured; an imaging section that images the cells, a lid section disposed on the main surface so as to cover the plurality of wells , a cell culture section, and an imaging section, are relatively moved in a direction along the main surface. a moving part , the cell culture part is arranged so that the main surface thereof is along the vertical direction, and the cell culture part is arranged in the concave well so that the main surface of the cell culture part is arranged along the vertical direction. The imaging section is placed so as to be recessed vertically downward, and the imaging section is relatively moved by the moving section with its main surface along the vertical direction. It is configured to image the state of cells cultured in the wells from a direction orthogonal to the main surface.

A cell culture imaging method according to a second aspect of the present invention includes a plate-shaped cell culture section having a main surface on which a plurality of concave wells are provided, and a lid section on the main surface so as to cover the plurality of wells. , a step of culturing cells with the main surface arranged along the vertical direction so that the concave well is recessed vertically downward; and a step of culturing the cells so that the main surface runs along the vertical direction. The state of the cells cultured in the wells of the cell culture unit is measured relative to the main surface while the cell culture unit and the imaging unit that images the cells cultured in the wells are moved relative to each other. and capturing images from orthogonal directions that intersect with each other. Further, a cell culture imaging device according to a third aspect of the present invention includes a plate-shaped cell culture section having a main surface provided with a plurality of concave wells in which cells are cultured, and a cell culture imaging device that images cells cultured in the wells. The cell culture section includes an imaging section and a lid section disposed on the main surface so as to cover the plurality of wells, the cell culture section is arranged so that the main surface is along the vertical direction, and the concave wells are approximately cylindrical. The concave well has a shape such that the curved surface of the approximately cylindrical concave part is recessed vertically downward while the cell culture part is arranged so that the main surface of the cell culture part is along the vertical direction. The imaging unit is configured to image the state of the cells cultured in the well from a direction perpendicular to the main surface with the main surface disposed along the vertical direction. ing.

According to the present invention, as described above, the imaging section detects the state of the cells cultured in the wells of the cell culture section, the main surface of which is arranged along the vertical direction, in a manner that intersects the main surface. It is configured to capture images from orthogonal directions. This allows cells to be imaged from a direction perpendicular to the direction along which gravity is applied (vertical direction), making it possible to image how cells grow in the vertical direction. . For example, it is possible to image a state in which a plurality of cells proliferate and a cluster of cells grows upward. Furthermore, in a cell cluster, dead cells are moved to the outside of the cluster. When this situation is imaged from below, a state in which dead cells are moved to the outer periphery of the aggregate is imaged. On the other hand, by imaging from the orthogonal direction intersecting the principal plane, it can be observed that dead cells move toward the outer periphery and downward side of the cell cluster. Note that the "orthogonal direction that intersects the main surface" includes a direction that intersects the main surface at an angle of 90 degrees and a direction that intersects the main surface at an angle near 90 degrees.

FIG. 1 is a diagram showing the configuration of a cell culture device according to one embodiment. FIG. 2 is a plan view of a cell culture unit according to one embodiment. 3 is a cross-sectional view taken along line 200-200 in FIG. 2. FIG. It is a perspective view (partially enlarged view) showing a cell culture part. It is a side view which shows the cell culture part of the state supported by the support part. It is a top view which shows the cell culture part of the state supported by the support part. FIG. 2 is a flow diagram illustrating a cell culture imaging method according to one embodiment. FIG. 2 is a diagram (side view) showing grown cells. FIG. 2 is a diagram (bottom view) showing grown cells. It is a perspective view showing the composition of the cell culture part by a modification. It is a sectional view showing the composition of the cell culture part by a modification.

DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments embodying the present invention will be described based on the drawings. In the following description, "approximately" is a concept that includes the shape (the direction) itself and approximately the shape (the direction).

(Configuration of cell culture imaging device)
The configuration of the cell culture imaging device 100 according to this embodiment will be described with reference to FIGS. 1 to 9. Note that in this specification, the "vertical direction" means a direction along the direction in which gravity is applied (Z direction). Further, directions along horizontal planes that are orthogonal to each other are defined as an X direction and a Y direction.

As shown in FIG. 1, the cell culture imaging device 100 includes a cell culture section 10 for culturing cells 1. The cell culture section 10 has a substantially plate shape (substantially rectangular shape). Further, the cell culture section 10 is arranged so that a main surface 11, which will be described later, extends along the vertical direction (Z direction).

As shown in FIGS. 2 and 3, the cell culture section 10 is plate-shaped and has a main surface 11 provided with a plurality of concave wells 12 in which cells 1 are cultured. Further, the plurality of wells 12 are arranged in a matrix on the main surface 11. The distance (pitch) between the centers of adjacent wells 12 is, for example, 700 μm. Further, the plurality of wells 12 are connected to each other by a flow path 13. Furthermore, a culture solution inlet 14 is provided at one end of the flow path 13 . Furthermore, an outlet 15 for the culture solution is provided at the other end of the flow path 13 . This makes it possible to culture the cells 1 in the wells 12 while flowing the culture medium through the flow channels 13.

Further, as shown in FIG. 4, the well 12 has a substantially cylindrical shape. The well 12 has a substantially circular shape when viewed from the Y direction. For example, the diameter of the well 12 when viewed from the Y direction is 400 μm. Note that the well 12 may have a size within a range of about 100 μm to about 1000 μm. Then, with the main surface 11 of the cell culture section 10 arranged such that the main surface 11 is along the vertical direction, the curved surface 12a of the approximately cylindrical recess is depressed vertically downward. It is set up so that As a result, the cells 1 gather together on the surface 12a located on the downward side, forming one aggregate (spheroid). Furthermore, the inner wall of the well 12 is coated with a polymer that is non-adhesive to the cells 1. Further, the concave well 12 is provided so as to be depressed vertically downward in a state where the cell culture section 10 is arranged such that the main surface 11 of the cell culture section 10 is along the vertical direction.

Further, as shown in FIG. 1, the cell culture imaging device 100 includes an imaging section 20. The imaging unit 20 is configured to image (magnify and image) the cells 1 cultured in the well 12. Further, the imaging section 20 includes an objective lens section 21, a lens barrel section 22, an imaging camera section 23, and a light source section 24. That is, the imaging unit 20 is composed of a microscope.

Here, in this embodiment, the imaging unit 20 is configured to image the state of the cells 1 cultured in the well 12 from a direction perpendicular to the main surface 11 (Y direction in this embodiment). has been done. Specifically, the objective lens section 21 and the lens barrel section 22 are arranged along the Y direction. Further, the objective lens section 21 and the lens barrel section 22 are arranged on the Y1 direction side with respect to the cell culture section 10. Further, the light source section 24 is arranged so that its optical axis runs along the Y direction. Further, the light source section 24 is arranged on the Y2 direction side with respect to the cell culture section 10 and so as to face the objective lens section 21. Further, the light source section 24 is made of, for example, an LED light source. The imaging camera section 23 is configured to capture an image of the cell 1 magnified by the objective lens section 21 from the Y1 direction side. Note that the "orthogonal direction that intersects the main surface 11" refers to a direction that intersects the main surface 11 at an angle of 90 degrees, and a direction that intersects the main surface 11 at an angle of approximately 90 degrees. include.

Furthermore, in this embodiment, the imaging unit 20 is configured to image the state of the cell 1 placed on the curved surface 12a of the substantially cylindrical recess from the Y direction. That is, the cells 1 (spheroids) are arranged on the curved surface 12a of the well 12 (approximately cylindrical recess). Cell 1 (spheroid) grows horizontally and vertically over time. The imaging unit 20 can image the cells 1 (spheroids) that grow to spread in the horizontal direction (mainly in the X direction) and the cells 1 (spheroids) that grow to spread in the vertical direction.

Moreover, in this embodiment, as shown in FIG. 1, the cell culture imaging device 100 includes a lid part 30. The lid part 30 is arranged on the main surface 11 so as to cover the plurality of wells 12. Further, the lid portion 30 is placed in a pressed state (in close contact with the main surface 11). The imaging unit 20 is configured to image the state of the cells 1 cultured in the well 12 from the Y direction with the lid 30 pressed against the main surface 11. Further, the lid portion 30 has a substantially plate shape (substantially rectangular shape). Further, all of the plurality of wells 12 are covered by one lid part 30.

Furthermore, in this embodiment, as shown in FIGS. 5 and 6, the cell culture imaging device 100 includes a support section 40. The support section 40 is provided so as to sandwich the cell culture section 10 and the lid section 30, with the lid section 30 disposed on the main surface 11. The support section 40 is configured to support the cell culture section 10. Specifically, the support section 40 has a substantially plate shape that covers the cell culture section 10 and the lid section 30. The size of the support part 40 (width W1 in the X direction and width W2 in the Z direction) is larger than the size (width W11 in the X direction and width W12 in the Z direction) of the cell culture part 10 and the lid part 30. .

Moreover, a pair of support parts 40 are provided. One of the pair of support parts 40 is arranged on the Y1 direction side of the cell culture section 10, and the other of the pair of support parts 40 is arranged on the Y2 direction side of the lid part 30. Then, the pair of support parts 40 are fixed to each other by bolts 41 with the cell culture part 10 and the lid part 30 being sandwiched between the pair of support parts 40 . Thereby, the lid section 30 is pressed against the cell culture section 10.

Furthermore, in this embodiment, the cell culture section 10 and the lid section 30 are each made of transparent members. For example, the cell culture section 10 and the lid section 30 are made of resin (for example, silicone rubber). Furthermore, the support section 40 is also made of a transparent member. For example, the support portion 40 is made of polycarbonate. Note that the "transparent member" is a member that allows light to pass through to the extent that the cell 1 placed in the well 12 can be imaged by the imaging unit 20 when the cell 1 is imaged from the Y1 direction side (or Y2 direction side). means a member that can

Furthermore, in this embodiment, as shown in FIG. 1, the cell culture imaging device 100 includes a moving section 50. The moving section 50 is configured to relatively move the cell culture section 10, which is arranged so that the main surface 11 is along the vertical direction, and the imaging section 20, in a direction along the main surface 11. There is. Specifically, the moving unit 50 includes a moving stage that moves the cell culture unit 10 in the Z direction and the X direction. For example, the moving unit 50 is constructed by tilting an XY stage by 90 degrees. The imaging unit 20 is configured to image the state of the cells 1 cultured in the wells 12 of the cell culture unit 10 while being relatively moved by the moving unit 50 from the Y direction. Note that in this embodiment, the imaging section 20 is stationary, and the cell culture section 10 is moved by the moving section 50.

Further, in the present embodiment, the imaging unit 20 scans the cells 1 cultured in the plurality of wells 12 arranged in a matrix of the cell culture unit 10 while being relatively moved by the moving unit 50. However, the camera is configured to capture images from the Y direction. For example, the cell culture unit 10 is moved along the Y1 direction with respect to the imaging unit 20 by the moving unit 50, and then moved in the Z2 direction. Thereafter, the cell culture section 10 is moved along the Y2 direction with respect to the imaging section 20 again. In this way, the imaging section 20 is scanned by moving the cell culture section 10 in a zigzag manner (meandering) relative to the imaging section 20.

Further, in the present embodiment, the imaging unit 20 images the cells 1 cultured in the well 12 from the Y direction at predetermined time intervals longer than the time required for one scan by the imaging unit 20. It is configured. For example, the imaging unit 20 images the cells 1 cultured in the wells 12 by scanning the plurality of wells 12 arranged in a matrix. After that, no imaging is performed for a certain period of time. After that, the imaging unit 20 again images the cells 1 cultured in the wells 12 by scanning the plurality of wells 12 arranged in a matrix. Then, the above operation is repeated. As a result, a time-lapse image of the cell 1 is captured. Note that while the time-lapse images of the cells 1 are being captured, the cell culture section 10 remains arranged along the vertical direction. That is, while time-lapse imaging of the cells 1 is being performed, the posture of the cell culture unit 10 does not change.

Furthermore, in this embodiment, the imaging unit 20 is configured to be able to switch between a mode in which images are taken for each well 12 and a mode in which images are taken for each of a plurality of wells 12. Specifically, when the imaging unit 20 images each well 12, the objective lens unit 21 is arranged at a position relatively close to the well 12. Further, when the imaging unit 20 images each of the plurality of wells 12, the objective lens unit 21 is arranged at a position relatively far from the wells 12. In this way, the imaging section 20 is configured such that the position of the objective lens section 21 with respect to the well 12 can be changed.

Further, in this embodiment, the cell culture section 10 and the imaging section 20 are housed inside a constant temperature bath 60. Note that the moving unit 50 is also housed inside the constant temperature bath 60. That is, imaging of the cells 1 (time-lapse image imaging) is performed while the temperature around the cell culture section 10 is maintained constant.

Next, a cell culture imaging method will be described with reference to FIG.

As shown in FIG. 7, in step 101, cells 1 are cultured. Specifically, in a plate-shaped cell culture section 10 having a main surface 11 on which a plurality of concave wells 12 are provided, the main surface 11 is vertically disposed so that the concave wells 12 are recessed vertically downward. Cells 1 are seeded and cultured while being arranged along the direction. In addition, the culture solution flows in from the inlet 14 of the channel 13, and the culture solution flows out from the outlet 15. The inflow and outflow of the culture solution is performed continuously while the cells 1 are being cultured.

Next, in step 102, the state of the cells 1 cultured in the wells 12 is imaged (enlarged) from a direction perpendicular to the main surface 11 (in the present embodiment, the Y direction). Further, by scanning by the imaging unit 20, the cells 1 cultured in the plurality of wells 12 arranged in a matrix are imaged for each well 12 (or for each plurality of wells 12).

Next, in step 103, it is determined whether a predetermined time has elapsed since the previous imaging of the cell 1 (at the end of one scan by the imaging unit 20). In the case of no in step 103, the determination in step 103 is repeated until a predetermined time has elapsed. If YES in step 103, the process returns to step 102, and the imaging unit 20 images the cell 1 again. Note that the posture of the cell culture section 10 is not changed between step 102 and step 103. In other words, the cell culture section 10 remains arranged along the vertical direction.

Furthermore, steps 102 and 103 are repeated until the imaging of the cell 1 (time-lapse image imaging) is completed. Note that even while the operations in steps 102 and 103 are being performed, the cells 1 are continuously cultured.

Next, the state of the cell 1 imaged by the imaging unit 20 will be explained.

For example, as shown in FIG. 8, the cell culture imaging device 100 can image a state in which a plurality of cells 1 proliferate and a cluster of cells 1 grows upward (Z1 direction). Furthermore, in the cluster of cells 1, dead cells 1a are moved to the outside of the cluster. When this situation is imaged from below, as shown in FIG. 9, a state in which the dead cells 1a are moved to the outer periphery of the agglomerate is imaged. On the other hand, by imaging from the Y direction (lateral direction) like the cell culture imaging device 100, it is possible to image that the dead cells 1a are moved to the outer circumferential side of the aggregate of cells 1 and to the downward side.

(Effects of this embodiment)
In this embodiment, the following effects can be obtained.

In the present embodiment, as described above, the imaging section 20 detects the state of the cells 1 cultured in the wells 12 of the cell culture section 10 arranged so that the main surface 11 is along the vertical direction. The camera is configured to take images from an orthogonal direction intersecting with the object. As a result, the cell 1 is imaged from a direction perpendicular to the direction along the direction of gravity (vertical direction), so it is possible to image how the cell 1 grows in the vertical direction. I can do it.

Further, in this embodiment, as described above, the concave well 12 has a substantially cylindrical shape. In the concave well 12, the curved surface 12a of the approximately cylindrical concave portion is directed vertically downward in a state in which the cell culture section 10 is arranged such that the main surface 11 of the cell culture section 10 is along the vertical direction. It is set up in a recessed manner. The imaging unit 20 is configured to image the state of the cell 1 placed on the curved surface 12a of the substantially cylindrical recess from an orthogonal direction. This makes it easier for a plurality of cells 1 to gather on the curved surface 12a of the substantially cylindrical recess, so that agglomerates (spheroids) of cells 1 are more likely to be generated. Therefore, by imaging the state of the cells 1 arranged on the curved surface 12a of the substantially cylindrical recess, the state of the aggregates (spheroids) of the cells 1 can be easily imaged.

Furthermore, in this embodiment, as described above, the cell culture imaging device 100 further includes the lid part 30 arranged on the main surface 11 so as to cover the plurality of wells 12. The imaging unit 20 is configured to image the state of the cells 1 cultured in the well 12 from an orthogonal direction with the lid 30 pressed against the main surface 11. As a result, the opening portion of the well 12 is covered while being pressed by the main surface 11, so even if the cell culture section 10 is arranged so that the main surface 11 is along the vertical direction, the cell culture section 10 from the well 12 It is possible to suppress the cells 1 and the culture solution from flowing out (leakage) to the outside of the cell.

Further, in this embodiment, as described above, the cell culture imaging device 100 is provided with the lid 30 disposed on the main surface 11 so as to sandwich the cell culture section 10 and the lid 30, and A support section 40 that supports the culture section 10 is further provided. The imaging unit 20 is configured to image the state of the cells 1 cultured in the wells 12 of the cell culture unit 10 supported by the support unit 40 from an orthogonal direction. As a result, even when the cell culture section 10 and the lid section 30 are made of a material that is relatively easily deformed (for example, silicone rubber), the cell culture section 10 and the lid section 30 are supported by the support section 40. Therefore, it is possible to suppress bending and deformation of the cell culture section 10 and the lid section 30.

Moreover, in this embodiment, the support part 40 has a substantially plate shape that covers the cell culture part 10 and the lid part 30, as described above. As a result, even if the cell culture section 10 and the lid section 30 are made of relatively soft materials, the cell culture section 10 and the lid section 30 are covered by the support section 40, so the cell culture section 10 and the lid section 30 are covered with the support section 40. The portion 30 can be easily arranged along the vertical direction (without being deformed). In addition, since the lid part 30 is reliably pressed against the wells 12 of the cell culture section 10 and the channels 13 that connect the wells 12, leakage of the culture solution etc. from the wells 12 and channels 13 is suppressed. can do.

Furthermore, in the present embodiment, as described above, the cell culture imaging device 100 connects the cell culture section 10, which is arranged so that the main surface 11 is along the vertical direction, and the imaging section 20 along the main surface 11. The apparatus further includes a moving part 50 that moves the moving part 50 relatively in the opposite direction. The imaging unit 20 is configured to image the state of the cells 1 cultured in the wells 12 of the cell culture unit 10 while being relatively moved by the moving unit 50 from an orthogonal direction. As a result, even when the plurality of wells 12 are arranged in a relatively wide range (wider than the field of view of the imaging unit 20), the cell culture unit 10 can be moved relative to the imaging unit 20 by the moving unit 50. Therefore, it is possible to image the cells 1 cultured in a plurality of wells 12 arranged in a relatively wide range.

Further, in this embodiment, as described above, the plurality of wells 12 are arranged in a matrix, and the imaging section 20 captures the cell culture section 10 that is relatively moved by the moving section 50. It is configured to scan and image cells 1 cultured in a plurality of wells 12 arranged in a matrix from orthogonal directions. As a result, the plurality of wells 12 arranged in a matrix are scanned by the imaging unit 20, so that all cells 1 cultured in the plurality of wells 12 arranged in a matrix are imaged (without omission). be able to.

Further, in the present embodiment, as described above, the imaging unit 20 collects the cells 1 cultured in the well 12 in the orthogonal direction at predetermined time intervals longer than the time required for one scan by the imaging unit 20. It is configured to take images from. Thereby, time-lapse imaging in which the cell 1 is imaged at regular time intervals can be easily performed.

Furthermore, in this embodiment, as described above, the imaging unit 20 is configured to be able to switch between a mode in which images are taken for each well 12 and a mode in which images are taken for each of a plurality of wells 12. As a result, if you want to image cells 1 with relatively high resolution (if you want to observe individual cells 1), you can image cells 1 for each well 12, and cells 1 cultured in multiple wells 12. When it is desired to compare the states of cells 1 in one image, it is possible to image the cells 1 for each of a plurality of wells 12. As a result, it is possible to capture an image according to the user's purpose.

Furthermore, in this embodiment, the cell culture section 10 and the imaging section 20 are housed inside the thermostatic bath 60, as described above. Thereby, the state of the cells 1 can be imaged while culturing the cells 1 inside the thermostatic bath 60 for a relatively long time. Furthermore, since there is no need to take out the cell culture unit 10 from the thermostatic bath 60 every time an image is taken, it is possible to suppress adverse effects on the cells 1 due to movement of the cell culture unit 10.

Furthermore, in this embodiment, as described above, the cell culture section 10 and the lid section 30 are each made of transparent members. Thereby, in a state where the cell culture section 10 is covered by the lid section 30, the light source section 24 is arranged on one side of the cell culture section 10 side and the lid section 30 side, and the imaging camera section 23 is arranged on the other side. With this, the cell 1 can be clearly imaged.

In addition, in the present embodiment, as described above, in the cell culture imaging method, in the plate-shaped cell culture section 10 having the main surface 11 in which a plurality of concave wells 12 are provided, the concave wells 12 are located on the vertically downward side. Step 101 of culturing the cells 1 in a state where the main surface 11 is disposed along the vertical direction so as to be recessed in the well 12, and the state of the cells 1 cultured in the well 12 relative to the main surface 11. and a step 102 of enlarging and capturing an image from orthogonal directions intersecting with each other. As a result, the cell 1 is imaged from a direction that intersects the direction along the direction of gravity (vertical direction), so it is possible to image how the cell 1 grows in the vertical direction. A possible cell culture imaging method can be provided.

(Modified example)
Note that the embodiments disclosed this time should be considered to be illustrative in all respects and not restrictive. The scope of the present invention is indicated by the claims rather than the description of the embodiments described above, and further includes all changes (modifications) within the meaning and range equivalent to the claims.

For example, in the embodiment described above, an example is shown in which the cell culture section 10 has a substantially plate shape, but the present invention is not limited to this. Like the cell culture section 110 according to the modified example shown in FIGS. 10 and 11, the cell culture section 110 may be composed of a microarray tape 110a that is flexible and can be wound in a circumferential shape. Specifically, the microarray tape 110a is formed into a band shape. Further, a plurality of wells 112 are provided in the microarray tape 110a. Moreover, when culturing the cells 1, the microarray tape 110a is wound.

In the cell culture unit 110 according to the modified example, when imaging the cells 1, the microarray tape 110a is at least partially unwound (flattened), and the unwound portion is imaged. be done. Note that during imaging, the microarray tape 110a is arranged so that the central axis C of the winding is along the vertical direction. Furthermore, in order to prevent the culture medium and cells 1 from leaking from the wells 112 even when the microarray tape 110a is unwound, a sheet-like sheet is placed on the main surface 111 of the microarray tape 110a where the wells 112 are formed. A lid portion 130 is provided. Further, the microarray tape 110a and the lid portion 130 are made of a flexible and transparent member (such as silicone rubber).

Further, in the above embodiment, an example was shown in which the well 12 is constituted by a substantially cylindrical recess (circular shape when viewed from the Y direction), but the present invention is not limited to this. For example, it is possible to apply the present invention to a well configured with a concave portion having a shape other than a circular shape (such as a substantially U-shape) when viewed from the Y direction.

Further, in the above embodiment, an example is shown in which the support section 40 that supports the cell culture section 10 is provided, but the present invention is not limited to this. For example, if the cell culture section and the lid are made of a relatively hard material (such as glass), the support section should not be provided (i.e., unless the cell culture section and the lid are bent and deformed). It is also possible.

Further, in the above embodiment, an example was shown in which the support section 40 has a substantially plate shape that covers the cell culture section 10 and the lid section 30, but the present invention is not limited to this. For example, if the cell culture section and the lid are made of a relatively hard material (such as glass), the support section may have a frame shape (that is, a shape that covers the outer edges of the cell culture section and the lid without covering the center part). ).

Further, in the embodiment described above, an example was shown in which the moving section 50 that moves the cell culture section 10 relative to the imaging section 20 is provided, but the present invention is not limited to this. For example, if the number of wells to be imaged is relatively small and all of the wells can be imaged without moving the imaging section, there is no need to provide a moving section.

[Mode]
It will be appreciated by those skilled in the art that the exemplary embodiments described above are specific examples of the following aspects.

(Item 1)
a plate-shaped cell culture section (10, 110) having a main surface (11, 111) provided with a plurality of concave wells (12, 112) in which cells 1 are cultured;
comprising an imaging unit 20 that images the cells 1 cultured in the wells (12, 112),
The cell culture section (10, 110) is arranged such that the main surface (11, 111) is along the vertical direction,
The concave well (12, 112) has the cell culture section (10, 110) arranged such that the main surface (11, 111) of the cell culture section (10, 110) is along the vertical direction. It is recessed vertically downward,
The imaging unit 20 is configured to image the state of the cells 1 cultured in the well (12, 112) from a direction orthogonal to the main surface (11, 111). Imaging device 100.

(Item 2)
The concave well (12, 112) has a substantially cylindrical shape,
The concave well (12, 112) is formed in a state where the cell culture section (10, 110) is arranged such that the main surface (11, 111) of the cell culture section (10, 110) is along the vertical direction. , the curved surface of the substantially cylindrical recess is recessed vertically downward;
The cell culture imaging device 100 according to item 1, wherein the imaging unit 20 is configured to image the state of the cells 1 placed on the curved surface of the substantially cylindrical recess from the orthogonal direction.

(Item 3)
further comprising a lid part 30 disposed on the main surface (11, 111) so as to cover the plurality of wells (12, 112),
The imaging unit 20 is configured to image the state of the cells 1 cultured in the wells (12, 112) from the orthogonal direction while the lid unit 30 is pressed against the main surface (11, 111). The cell culture imaging device 100 according to item 1 or 2, configured as follows.

(Item 4)
The cell culture section (10, 110) is provided so as to sandwich the cell culture section (10, 110) and the cover section 30 with the lid section 30 disposed on the main surface (11, 111). ), further comprising a support part 40 that supports the
The imaging unit 20 is configured to image the state of the cells 1 cultured in the wells (12, 112) of the cell culture unit (10, 110) supported by the support unit 40 from the orthogonal direction. The cell culture imaging device 100 according to item 3, configured as follows.

(Item 5)
The cell culture imaging device 100 according to item 4, wherein the support section 40 has a substantially plate shape that covers the cell culture section (10, 110) and the lid section 30.

(Item 6)
The cell culture section (10, 110), in which the main surface (11, 111) is arranged along the vertical direction, and the imaging section 20 are arranged in the direction along the main surface (11, 111). , further comprising a moving unit 50 for relatively moving,
The imaging unit 20 detects the state of the cells 1 cultured in the wells (12, 112) of the cell culture unit (10, 110) while being relatively moved by the moving unit 50 in the orthogonal direction. The cell culture imaging device 100 according to any one of items 1 to 5, configured to take images from.

(Item 7)
The plurality of wells (12, 112) are arranged in a matrix,
The imaging unit 20 is configured to perform culture in the plurality of wells (12, 112) arranged in a matrix of the cell culture unit (10, 110), which are relatively moved by the moving unit 50. The cell culture imaging device 100 according to item 6, wherein the cell culture imaging device 100 is configured to image the cell 1 from the orthogonal direction while scanning it.

(Item 8)
The imaging unit 20 is configured to image the cells 1 cultured in the wells (12, 112) from the orthogonal direction at predetermined time intervals longer than the time required for one scan by the imaging unit 20. The cell culture imaging device 100 according to item 7, configured as follows.

(Item 9)
Item 1, wherein the imaging unit 20 is configured to be capable of imaging by switching between a mode of imaging for each well (12, 112) and a mode of imaging for each of a plurality of wells (12, 112). The cell culture imaging device 100 according to any one of items 1 to 8.

(Item 10)
The cell culture imaging device 100 according to any one of items 1 to 9, wherein the cell culture section (10, 110) and the imaging section 20 are housed inside a constant temperature bath 60.

(Item 11)
further comprising a lid part 30 disposed on the main surface (11, 111) so as to cover the plurality of wells (12, 112),
The cell culture imaging device 100 according to any one of items 1 to 10, wherein the cell culture section (10, 110) and the lid section 30 are both made of transparent members.

(Item 12)
In a plate-shaped cell culture section (10, 110) having a main surface (11) on which a plurality of concave wells (12, 112) are provided, the concave wells (12, 112) are depressed vertically downward. a step of culturing the cells 1 in a state where the main surfaces (11, 111) are arranged along the vertical direction so as to be provided;
A cell culture imaging method comprising the step of imaging the state of cells 1 cultured in the wells (12, 112) from a direction perpendicular to the main surface (11, 111).

1 Cells 10, 110 Cell culture section 11, 111 Main surface 12, 112 Well 12a Surface 20 Imaging section 30, 130 Lid section 40 Support section 50 Moving section 60 Constant temperature bath 100 Cell culture imaging device

Claims (14)

a plate-shaped cell culture section having a main surface provided with a plurality of concave wells in which cells are cultured;
an imaging unit that images the cells cultured in the well;
a lid disposed on the main surface so as to cover the plurality of wells;
a moving unit that relatively moves the cell culture unit and the imaging unit in a direction along the main surface,
The cell culture section is arranged such that the main surface is along a vertical direction,
The concave well is provided so as to be recessed vertically downward in a state where the cell culture section is arranged such that the main surface of the cell culture section is along the vertical direction,
The imaging unit detects the state of the cells cultured in the well of the cell culture unit in a state where the main surface is arranged along the vertical direction and is relatively moved by the moving unit . A cell culture imaging device configured to capture images from an orthogonal direction intersecting the main surface. The concave well has a substantially cylindrical shape,
The concave well is provided such that the curved surface of the substantially cylindrical concave portion is depressed vertically downward in a state in which the cell culture section is arranged such that the main surface of the cell culture section is along the vertical direction. has been
The cell culture imaging device according to claim 1, wherein the imaging unit is configured to image the state of cells arranged on the curved surface of the substantially cylindrical recess from the orthogonal direction. The imaging unit is configured to image the state of the cells cultured in the well from the orthogonal direction while the lid is pressed against the main surface. cell culture imaging device. further comprising a support part that is provided to sandwich the cell culture part and the lid part with the lid part disposed on the main surface, and supports the cell culture part;
The cell according to claim 3, wherein the imaging unit is configured to image the state of the cells cultured in the well of the cell culture unit supported by the support unit from the orthogonal direction. Culture imaging device. The cell culture imaging device according to claim 4, wherein the support section has a substantially plate shape that covers the cell culture section and the lid section. The plurality of wells are arranged in a matrix,
The imaging unit scans the cells cultured in the plurality of wells arranged in a matrix of the cell culture unit, which are relatively moved by the moving unit, from the orthogonal direction. The cell culture imaging device according to any one of claims 1 to 5 , configured to take an image. The imaging unit is configured to image the cells cultured in the well from the orthogonal direction at predetermined time intervals longer than the time required for one scan by the imaging unit. 6. The cell culture imaging device according to 6 . The imaging unit is configured to be capable of imaging by switching between a mode of imaging for each well and a mode of imaging for each of a plurality of wells. Cell culture imaging device. The cell culture imaging device according to any one of claims 1 to 8 , wherein the cell culture section and the imaging section are housed inside a constant temperature bath. The cell culture imaging device according to any one of claims 1 to 9 , wherein the cell culture section and the lid section are each made of transparent members. The cell culture imaging device according to any one of claims 1 to 10 , wherein the cell culture section has an inlet through which a culture solution flows into the well and an outlet through which a culture solution flows out from the well. The cell culture imaging device according to any one of claims 1 to 11 , wherein the inner wall of the well is coated with a polymer that is non-adhesive to the cells. In a plate-shaped cell culture section having a main surface on which a plurality of concave wells are provided, and a lid part is arranged on the main surface so as to cover the plurality of wells, the concave wells are arranged vertically downward. culturing the cells in a state where the main surface is arranged along the vertical direction so as to be recessed;
The cell culture section is moved relative to the cell culture section and an imaging section that images the cells cultured in the well, with the main surface arranged along the vertical direction. A cell culture imaging method comprising the step of imaging the state of cells cultured in the well from an orthogonal direction intersecting the main surface. a plate-shaped cell culture section having a main surface provided with a plurality of concave wells in which cells are cultured;
an imaging unit that images the cells cultured in the well;
a lid disposed on the main surface so as to cover the plurality of wells,
The cell culture section is arranged such that the main surface is along a vertical direction,
The concave well has a substantially cylindrical shape,
The concave well is configured such that the curved surface of the substantially cylindrical concave portion is depressed vertically downward in a state in which the cell culture section is arranged such that the main surface of the cell culture section is along the vertical direction. It is provided,
The imaging unit is configured to image the state of the cells cultured in the well from a direction orthogonal to the main surface, with the main surface disposed along the vertical direction. A cell culture imaging device.

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