JP7130396B2 - Analysis device, analysis system, program - Google Patents

Description

The present invention relates to an analysis device, an analysis system , and a program in a culture system.

In cell culture, suspension culture is sometimes performed in which cells are cultured while being suspended in a culture solution and stirred. In suspension culture, information such as the number of cells or the size of cell clusters may be required. For example, Patent Literature 1 discloses a technique related to a culture state analysis system that calculates the particle size distribution of cell aggregates in a suspension culture medium and the total number of particles of cell aggregates in the entire suspension culture vessel. In this system, a specific area in a suspension culture medium containing a plurality of cell aggregates with different particle sizes is imaged. The state of the cell mass is estimated using the information obtained from the photographed image and known information. Based on the results, the particle size distribution and total particle number of the cell mass are calculated. In suspension culture, it is meaningful to obtain information about the state of cells.

Japanese Patent Application Laid-Open No. 2017-140006

An object of the present invention is to provide an analysis system and the like used in a culture system for performing suspension culture.

According to one aspect of the invention, an analysis system comprises a camera and an analysis device having a processor. The camera is arranged on the side of a spinner flask in which cells are cultured in suspension in a culture medium, and is configured to repeatedly photograph the spinner flask to obtain a group of images. A processor extracts from the images a first image taken when the blades of the spinner flask are along the optical axis of the camera, and based on the obtained first image , configured to analyze at least one of the morphology and size of the cells .

ADVANTAGE OF THE INVENTION According to this invention, the analysis system etc. which are used for the culture system for performing floating culture can be provided.

FIG. 1 is a diagram showing an outline of a configuration example of a culture system. FIG. 2 is a block diagram showing an outline of a configuration example of a camera and a controller in the culture system according to the first embodiment; FIG. 3 is a flowchart outlining an example of the operation of the controller according to the first embodiment; FIG. 4A is a diagram for explaining the positional relationship between the blades of the spinner flask and the camera when acquiring the first image. FIG. 4B is a diagram for explaining the positional relationship between the blades of the spinner flask and the camera when acquiring the second image. FIG. 5 is a diagram for explaining the positional relationship between the blades of the spinner flask and the camera according to the second modification. FIG. 6 is a diagram for explaining the outline of the shape of the blades of the spinner flask according to the third modification. FIG. 7 is a flowchart outlining an example of the operation of the culture system according to the second embodiment. FIG. 8 is a flowchart outlining an example of the operation of the controller according to the second embodiment. FIG. 9 is a diagram schematically showing an example of a menu screen displayed on a controller according to the second embodiment; FIG. 10 is a flowchart outlining an example of camera setting processing performed by the controller according to the second embodiment. FIG. 11 is a flowchart showing an outline of an example of measurement processing performed by the controller according to the second embodiment. 12A is a diagram schematically showing an example of a measurement screen displayed on a controller according to the second embodiment; FIG. FIG. 12B is a diagram showing an example of a three-dimensional bar graph representing the relationship between the size of cell clusters, the floating position, and the number of cell clusters. FIG. 12C is a diagram showing an example of a bubble map representing the relationship between the size of cell clusters, floating positions, and the number of cell clusters. FIG. 12D is a diagram showing an example of a heat map representing the relationship between the size of cell clusters, floating positions, and the number of cell clusters. FIG. 13 is a flowchart outlining an example of the operation of the camera according to the second embodiment. FIG. 14 is a block diagram showing an outline of a configuration example of a culture system according to the third embodiment. FIG. 15A is a flowchart outlining an example of the operation of the culture system according to the third embodiment; FIG. 15B is a flowchart outlining an example of the operation of the culture system according to the third embodiment; FIG. 15C is a flowchart outlining an example of the operation of the culture system according to the third embodiment; FIG. 16 is a diagram schematically showing an example of a menu screen displayed on a controller according to the third embodiment; FIG. 17A is a diagram for explaining changes in the amount of light over time when the camera position is appropriate. FIG. 17B is a diagram for explaining changes in the amount of light over time when the camera position is not appropriate. FIG. 18 is a diagram schematically showing an example of a control screen displayed on a controller according to the third embodiment; FIG. 19 is a flowchart outlining an example of the operation of the controller according to the third embodiment; FIG. 20 is a flowchart outlining an example of observation position setting processing performed by the controller according to the third embodiment. FIG. 21 is a flowchart showing an outline of an example of measurement setting processing performed by a controller according to the third embodiment; FIG. 22 is a flow chart showing an outline of an example of the operation of the strut according to the third embodiment. FIG. 23 is a flow chart showing an outline of an example of the operation of the stirrer according to the third embodiment.

[First Embodiment]
The first embodiment relates to a system for floating culture of cells or the like in a liquid culture medium. Suspension culture generally does not require enzymatic or mechanical dissociation of cells from a culture container, which is performed in the case of adherent culture, and is easy to subculture. In addition, since the number of cells is not limited by the surface area of the container as in the case of adherent culture, and the cells can be cultured three-dimensionally in the culture medium, it is easy to scale up the culture. Therefore, suspension culture is used in many applications such as obtaining a large amount of cultured cells and obtaining a large amount of protein from a large amount of cells. Suspension culture requires continuous adequate agitation of the culture medium for sufficient gas exchange. Also, in suspension culture, it is necessary to timely count the number of cells in order to track the growth pattern. In suspension culture, cell growth can be promoted by agitating cells at an appropriate concentration. In suspension culture, information such as the number of cells or the size of cell clusters can be useful information.

The culture system of this embodiment acquires an image of a sample undergoing suspension culture using a camera. The culture system analyzes the culture conditions, which may include cell density, cell morphology, etc., based on the obtained images. It can be said that the culture system is equipped with an analysis device that acquires the state of culture. A culture system can be said to include an analysis system having an analysis device.

<System configuration>
FIG. 1 shows an outline of a configuration example of a culture system 10 according to this embodiment. The culture system 10 includes a controller 100 , a camera 200 , a stirrer 300 and a spinner flask 400 . A sample 500 containing a culture medium, cells, etc. is placed inside a spinner flask 400 .

The spinner flask 400 has a cylindrical container body 402 . Here, the cylindrical shape means an approximately cylindrical shape, and does not need to be strictly cylindrical. A lid 404 is provided on the top of the container body 402 . An upper end of a rotary shaft 412 is rotatably attached to the central portion of the lid 404 inside the container body 402 . The rotary shaft 412 is arranged to extend downward along the central axis of the cylindrical container body 402 . A stirrer bar 452 is fixed to the lower end of the rotating shaft 412 . Moreover, a plurality of blades 420 are provided radially around the rotating shaft 412 on the upper side of the stirrer bar 452 of the rotating shaft 412 . The vane 420 is provided so as to be submerged in the sample 500 placed in the container body 402 . The vane 420 preferably reflects illumination light when the sample 500 is imaged by the camera 200 . Therefore, the vanes 420 are, for example, white or mirror-finished.

During culture, the spinner flask 400 containing the sample 500 is placed on the stirrer 300, which will be described later. A stirrer bar 452 in the container body 402 is rotated by the stirrer 300 . Along with this, the blades 420 inside the container body 402 rotate around the rotation shaft 412 . This rotation causes the blades 420 to agitate the sample 500 .

The configuration of spinner flask 400 shown here is an example. Spinner flask 400 may take other forms with similar functionality.

Stirrer 300 comprises magnet plate 352 and motor 354 . Magnet plate 352 is a rotating plate with magnets for rotating the stirrer bar on stirrer 300 . A motor 354 rotates the magnet plate 352 . A rotating shaft of the motor 354 is connected to the center of the magnet plate 352 .

The stirrer 300 also includes a third input device 332 and a third display device 334 . The third input device 332 includes a switch for switching the presence/absence of rotation of the magnet plate 352, a knob for adjusting the rotation speed of the magnet plate 352, and the like. A third display device 334 displays information such as the presence or absence of rotation of the magnet plate 352 and the rotation speed. The stirrer 300 also includes a third circuit group 305 including a control circuit for controlling the operation of each part of the stirrer 300 .

Camera 200 photographs sample 500 in spinner flask 400 . Camera 200 is positioned on the side of spinner flask 400 . The camera 200 includes a camera body 202 including an imaging device 260 and a lens unit 250 . The lens unit 250 includes a lens group that forms a subject image on the imaging device 260 . Camera 200 includes an illumination unit 280 that emits illumination light for illuminating a subject. The camera body 202 includes a second circuit group 205 including control circuits and the like for controlling the operation of each part of the camera 200 . Camera 200 is fixed to support 290 . At this time, the orientation, height, etc. of the camera 200 are adjusted so that the lens unit 250 faces the sample 500 in the spinner flask 400 .

In order to confirm the distribution of cells floating or sedimented in the spinner flask 400, it is necessary to photograph the spinner flask 400 widely in the vertical direction. In general, the imaging element 260 of the camera 200 is often rectangular. Therefore, it is preferable that the camera 200 is arranged such that the long side of the imaging device 260 is in the vertical direction. In addition, since it is sufficient if the spinner flask 400 can be photographed widely in the vertical direction, the camera 200 may be arranged so that the diagonal diagonal of the imaging device 260 is in the vertical direction. Furthermore, a plurality of cameras may be arranged in order to photograph a wide range.

In this embodiment, the XYZ axes are defined as follows. A horizontal Z axis is defined as the orientation of the optical axis of the lens unit 250 of the camera 200 . A Y-axis is defined vertically upward, ie parallel to the axis of rotation 412 of the spinner flask 400 . Define the X axis perpendicular to the Z and Y axes.

The controller 100 is an information processing device such as a personal computer (PC) or a tablet information terminal. FIG. 1 shows a tablet information terminal. Controller 100 communicates with camera 200 and acquires images from camera 200 . The controller 100 analyzes the acquired image and calculates necessary information.

The controller 100 comprises a first circuit group 105 including circuits such as processors that perform various calculations. The controller 100 also includes a touch screen 133 that functions as a display device and an input device. The controller 100 further includes an input button 136 as an input device. The controller 100 functions as an analysis device that analyzes the state of culture.

The configuration of the controller 100 and camera 200 will be further described with reference to the block diagram shown in FIG.

Controller 100 includes a first input device 132 and a first display device 134 . The first input device 132 includes a touch panel that is part of the touch screen 133, input buttons 136, and the like. The first display device 134 includes a display device or the like that is part of the touch screen 133 . This display device may be a liquid crystal display, an organic EL display, or the like.

The controller 100 comprises a first processor 110, a first storage circuit 116, and a first communication circuit 120, which may be included in the first circuit group 105 described above. The first processor 110, the first memory circuit 116, the first communication circuit 120, the first input device 132, and the first display device 134 are connected to each other via a first bus line 119. can be connected.

The first processor 110 includes an integrated circuit such as, for example, a Central Processing Unit (CPU), an Application Specific Integrated Circuit (ASIC), a Field Programmable Gate Array (FPGA), or a Graphics Processing Unit (GPU). The first processor 110 may be configured by one integrated circuit or the like, or may be configured by combining a plurality of integrated circuits or the like. The first processor 110 operates according to a program recorded in, for example, the first memory circuit 116 or a recording area within the first processor 110 .

The first storage circuit 116 includes a read only memory (ROM) that records a boot program and the like, a random access memory (RAM) that functions as a main storage device for the CPU, a storage, and the like. For RAM, for example, Dynamic RAM (DRAM), Static RAM (SRAM), etc. can be used. For storage, for example, Hard Disk Drive (HDD), Solid State Drive (SSD), Embedded Multi Media Card (eMMC), etc. are used. Various information such as programs and parameters used by the CPU are recorded in the storage.

A first communication circuit 120 is a circuit for communicating with the camera 200 . Communication with the camera 200 may be performed by wire or wirelessly. When performed wirelessly, a relatively high-speed communication method such as Wi-Fi (registered trademark), for example, can be used to transmit and receive images. A low power consumption communication method such as Bluetooth (registered trademark) may be used for transmitting and receiving data with a small amount of information such as an operation command.

The camera 200 includes a second input device 232 and a second display device 234 in addition to the lens unit 250 , image sensor 260 and illumination unit 280 described above. The second input device 232 is various devices for the user to input instructions to the camera 200 . The second input device 232 may include button switches, dials, touch panels, or the like. The second display device 234 is a display device for displaying the state of the camera 200, an image captured by the camera 200, or the like. The second display device 234 may include a liquid crystal display, pilot lamp, or the like.

The camera 200 includes a second processor 210, an image processing circuit 214, a second memory circuit 216, and a second communication circuit 220 included in the second circuit group 205 described above.

The second processor 210 includes an integrated circuit such as, for example, a CPU, ASIC, or FPGA. The second processor 210 may be configured by one integrated circuit or the like, or may be configured by combining a plurality of integrated circuits or the like. The second processor 210 operates according to a program recorded in, for example, the second storage circuit 216 or a recording area within the second processor 210 .

Image processing circuitry 214 may include a dedicated integrated circuit for image processing, such as an ASIC or GPU. The image processing circuit 214 may not be provided separately from the second processor 210 and the functions of the image processing circuit 214 may be performed by the second processor 210 .

The second storage circuit 216 includes necessary storage media such as ROM, RAM, and flash memory. Camera 200 may be configured such that a storage medium removable from camera 200 is connected.

A second communication circuit 220 is a circuit for communicating with the controller 100 . Communication between the controller 100 and the camera 200 may be performed by wire or wirelessly as described above. When performed wirelessly, a relatively high-speed communication method such as Wi-Fi can be used for transmitting and receiving images.

Second processor 210 , image processing circuit 214 , second memory circuit 216 , second communication circuit 220 , second input device 232 , second display device 234 , lens unit 250 , imaging device 260 , lighting unit 280 etc. are connected to each other via a second bus line 219 .

<System operation>
In the culture system 10 according to this embodiment, the stirrer 300 rotates the blades 420 of the spinner flask 400 at a preset rotation speed. Camera 200 continuously acquires images of sample 500 in spinner flask 400 . Camera 200 transmits the captured image to controller 100 . The controller 100 acquires continuous images captured by the camera 200 and performs various analyzes based on these images. The analysis performed by the controller 100 may include information such as the morphology and size of cells or cell clusters in the sample 500, the number or density of cells or cell clusters in the sample 500, and the pH of the culture solution.

The operation of culture system 10 will be described with reference to FIG. In step S<b>101 , the controller 100 acquires continuous images from the camera 200 . The sequential images are images of the sample 500 in the spinner flask 400 continuously captured by the camera 200 . A continuous image may be a moving image or a set of still images. In either case the sequence of images forms an image group comprising a plurality of images. In step S102, the controller 100 identifies the number of revolutions per unit time of the blades 420 of the spinner flask 400 based on the periodic changes in the obtained image.

In step S103, the controller 100 extracts the first image from the acquired continuous images. The first image is an image in which cells can be observed in a monolayer. Information on the number of revolutions of the identified blade 420 may also be used to extract the first image. At step S104, the controller 100 extracts a second image from the acquired continuous images. The second image is an image in which multilayered cells can be observed. Information on the number of revolutions of the identified blade 420 may also be used to extract the second image.

The first image and the second image will be described with reference to FIGS. 4A and 4B. 4A and 4B are diagrams schematically showing horizontal cross sections of the spinner flask 400 passing through the optical axis of the camera 200. FIG. In camera 200 , lens unit 250 attached to camera body 202 faces toward the center of spinner flask 400 . The spinner flask 400 has four blades, a first blade 421 , a second blade 422 , a third blade 423 and a fourth blade 424 fixed around a rotating shaft 412 . A sample 500 includes a culture medium 520 and cells 510 floating in the culture medium 520 . Here, the cell 510 can float in the culture solution 520 as a single cell or as a cell cluster in which a plurality of cells are gathered. FIG. 4A schematically illustrates the position of vane 420 relative to camera 200 when the first image is acquired. FIG. 4B schematically illustrates the position of vane 420 relative to camera 200 when the second image is acquired.

When the first image is acquired, one of the vanes 420 , the first vane 421 in the example of FIG. 4A, is positioned along the optical axis of the lens unit 250 . At this time, a relatively thin layer of culture solution 520 is formed between the end face 431 of the first blade 421 and the container body 402 . This layer has a thickness of, for example, about 5 mm. Therefore, the depth of culture medium 520 is limited to about 5 mm when viewed from camera 200 . Therefore, the cells in the culture solution 520 are restricted in their positions in the Z direction (optical axis direction of the lens unit 250). As a result, the camera 200 can image the cells under substantially uniform conditions, and can eliminate the error that cells appear small because they are far away. Therefore, the accuracy of analysis of the size of cells or cell clusters based on captured images is enhanced. Also, in this layer, one cell or cell cluster may exist in the optical axis direction of the lens unit 250 . In other words, in the optical axis direction of the lens unit 250, a state can occur in which cell clusters or the like exist in a single layer without overlapping. Cells 510 in such a state are referred to as monolayer cells 511 in this embodiment. In the state shown in FIG. 4A, camera 200 can acquire an image of monolayer cell 511 as the first image.

When the second image is acquired, the optical axis of the lens unit 250 is positioned between two of the vanes 420, in the example of FIG. 4B between the first vane 421 and the fourth vane 424. is doing. In other words, the first blade 421 and the fourth blade 424 sandwich the optical axis of the lens unit 250 . That is, none of the vanes 420 are aligned with the optical axis of the lens unit 250 . At this time, the camera 200 can see a relatively thick layer of the culture medium 520 surrounded by the main surface 432 of the first blade 421 and the main surface 432 of the fourth blade 424 and the container body 402 . In this thick culture medium layer, a plurality of cells or cell clusters may exist in the optical axis direction of the lens unit 250 . In other words, camera 200 can photograph a state in which many cells 510 are suspended in culture solution 520 . Such a sample 500 as viewed by camera 200 will be referred to as suspended sample 512 in this embodiment. When in a state such as that shown in FIG. 4B, an image of suspended sample 512 can be acquired as a second image.

In step S105, the controller 100 performs, for example, the following analysis based on the extracted first image. Controller 100 can analyze the morphology of monolayer cells 511 such as cell clusters based on the first image. Controller 100 can analyze the size of the cell mass based on the first image. Controller 100 can count the number of monolayer cells 511 per unit area and calculate the density of cells 510 based on the first image.

In step S106, the controller 100 performs, for example, the following analysis based on the extracted second image. Controller 100 can calculate the turbidity of sample 500 based on the second image and the density of cells 510 in suspended sample 512 based on the turbidity. Once the density of cells 510 is determined, the number of cells in sample 500 is also determined. The controller 100 can identify the color of the culture fluid 520 based on the second image, and calculate the pH value of the culture fluid 520 associated with this color based on the identified color. For example, if phenol red is added to the culture medium 520, if the pH value is appropriate, the culture medium 520 will appear red, and if the cells 510 increase and the culture medium 520 will become acidic, the culture medium 520 will turn yellow. Change. Controller 100 can analyze the state of the culture solution based on the color data of the second image. The controller 100 can calculate the pH value as the state of the culture solution. When calculating the pH value, a highly accurate value can be calculated by simple processing such as calculating an integrated value or an average value for a plurality of times for the color data of the second image and using the calculated value. Further, since the calculation of the pH value is based on the second image, when calculating the pH value, the culture vessel is not limited to a spinner flask that stirs the culture solution using blades, can be used.

In step S107, the controller 100 presents the analysis results and the like to the user using the touch screen 133 and the like. The presented analysis results and the like include the number of rotations per unit time of the blades 420 of the spinner flask 400, the first image, the second image, the feature amount of the cell mass, the density of the cells 510, and the cells based on the density of the cells. , the distribution of the number of particles for each characteristic value of the cell mass, the pH value of the culture solution 520, or the like. Also, the controller 100 records this information in the first memory circuit 116 . For example, the characteristic amount of a cell aggregate includes the form of the cell aggregate or the size of the cell aggregate, and examples of the form or size of the cell aggregate include the area, volume, average diameter, and maximum diameter of the cell aggregate.

As described above, in the culture system 10 according to this embodiment, the camera 200 continuously acquires images of the sample 500 inside the spinner flask 400 in which the blades 420 rotate. The controller 100 extracts two types of images, a first image and a second image, from the acquired consecutive images. The controller 100 performs image extraction based on the image obtained from the camera 200 without using the rotational speed of the stirrer 300 or the photographing conditions of the camera 200 . By photographing from the side of the spinner flask 400, analysis targeting floating cells can be performed. The controller 100 can acquire information such as the morphology of the cell mass, the size of the cell mass, the density of the cells 510, the number of cells, or the pH value of the culture medium 520 based on the extracted image.

According to this embodiment, the culture system 10 can monitor the condition of the sample 500 during culture. By using the culture system 10, the user can obtain information necessary for culture work, such as the timing of medium replacement and passage. The user can obtain information on the state of the sample 500 being analyzed, such as cell density, cell clump morphology, or cell clump size. These pieces of information are meaningful information for users conducting various experiments or cell cultures.

For example, in quality control in the production of stem cells such as iPS cells or ES cells, information such as the characteristic amount of cell aggregates and cell density plays an important role. The culture system 10 of this embodiment can easily acquire such information.

In addition, in the present embodiment, the case where the information regarding the shooting of the camera 200 is not acquired has been described as an example. On the other hand, the controller 100 can also acquire information regarding shooting, such as shooting conditions and shooting timing, from the camera 200 . The controller 100 may also use the shooting information acquired from the camera 200 to extract and analyze images. Moreover, in this embodiment, the case where both the first image and the second image are extracted and both images are used for analysis has been described. On the other hand, one of the first image and the second image may be extracted and the analysis may be performed using only the extracted image.

[First modification]
Regarding the first modified example, differences from the first embodiment will be described. Although the controller 100 is an information processing device such as a PC or a tablet-type information terminal in the above embodiment, it is not limited to this. Some or all of the functions of the controller 100 may be performed by a server on the network. Functions such as image analysis may be provided in the form of web services.

[Second modification]
Regarding the second modification, differences from the first embodiment will be described. In the above-described embodiment, as described with reference to FIG. 4A, the first image shows an example obtained when the optical axis of lens unit 250 of camera 200 and blade 420 are aligned in a straight line. rice field. In this case, the camera 200 captures a subject image of the monolayer cell 511 incident from the front of the imaging device 260 .

On the other hand, in this modified example, as shown in FIG. 5, the camera 200 is configured to capture a subject image that is obliquely incident on the optical axis of the lens unit 250 . More specifically, the camera 200 is oriented such that the optical axis of the lens unit 250 is offset from the rotation axis 412 of the spinner flask 400 . The illumination unit 280 irradiates illumination light from a direction oblique to the optical axis of the lens unit 250 so that the image of the monolayer cell 511 is obliquely incident on the lens unit 250 .

As in this example, the subject image is obliquely incident on the optical axis of the lens unit 250, so that the camera 200 acquires an image such as a differential interference contrast image that facilitates capturing the three-dimensional shape of the monolayer cells 511. can. It is significant that the first image showing the morphology of cell clusters and the like is acquired as an image such as a pseudo differential interference contrast image that makes it easy to capture the three-dimensional shape.

[Third Modification]
Regarding the third modification, differences from the first embodiment will be described. In the above-described embodiment, the first image is taken as an image of monolayer cells 511 at the end face 431 of the vanes 420 of the spinner flask 400, as described with reference to FIG. 4A. In this modified example, a flat plate 435 is provided at the end of the blade 420 of the spinner flask 400 so that such an image of the monolayer cell 511 can be easily acquired. It is preferable that the surface of the flat plate 435 facing the camera 200 has a characteristic of easily reflecting illumination light.

In other words, the spinner flask 400 of this modified example has a container body 402 having a cylindrical shape, blades 420 having a surface extending in the circumferential direction from the axis of the cylinder and rotating around the axis, and A flat plate 435 is provided and faces the circumferential surface of the cylindrical container body 402 . Note that the shape of the flat plate 435 is not limited to a flat plate. Any shape may be used as long as the plate is provided on the side of the circumferential surface of the blade 420 and has a surface facing the circumferential surface.

By providing a flat plate 435 wider than the end surface 431 at the end of the blade 420, the image of the monolayer cell 511 can be easily acquired in acquiring the first image. In addition, the blades are not limited to being provided with a plate, and it is sufficient if the blades are provided with surfaces at the ends thereof, for example, the blades 420 are thicker at the ends. Also, this surface may be provided obliquely to the circumferential surface instead of facing the circumferential surface.

[Second embodiment]
A second embodiment will be described. Here, the points of difference from the first embodiment will be described, and the same reference numerals will be given to the same parts, and the description thereof will be omitted. In the first embodiment, the camera 200 continuously acquires images independently of the controller 100 and transmits the acquired images to the controller 100 . The controller 100 performs various analyzes based only on the acquired images. In contrast, in this embodiment, the controller 100 controls the operation of the camera 200 .

An outline of the operation of the culture system 10 according to this embodiment will be described with reference to the flowchart shown in FIG. This flow chart shows the operation of the controller 100 and the operation of the camera 200 .

In step S201, the controller 100 requests the camera 200 for an image. In response to this request, in step S202, the camera 200 continuously takes pictures and transmits the obtained images to the controller 100. FIG.

In step S<b>203 , controller 100 acquires an image captured by camera 200 . The controller 100 analyzes the shooting based on the acquired image. More specifically, the rotation of the vanes 420 of the spinner flask 400 is analyzed to obtain information regarding when to take the first image and when to take the second image. In step S204, the controller 100 specifies imaging timings for acquiring the first image and the second image based on the analysis result. In step S<b>205 , the controller 100 transmits to the camera 200 information about the shooting conditions including the specified shooting timing.

In step S<b>206 , the camera 200 acquires information regarding the shooting conditions transmitted from the controller 100 . The camera 200 sets shooting conditions based on the acquired information. In step S207, the camera 200 performs a shooting operation based on the set shooting conditions. Thus, the camera 200 captures the first image and the second image. Camera 200 transmits the acquired first image and second image to controller 100 .

In step S<b>208 , controller 100 acquires a first image and a second image from camera 200 . The controller 100 performs various analyzes based on the obtained first and second images, as in the first embodiment. In step S<b>209 , the controller 100 presents the analysis results and the like to the user using the first display device 134 and stores them in the first storage circuit 116 .

Among the operations of the culture system 10 according to the present embodiment described above, the operation of the controller 100 will be described with reference to the flowchart shown in FIG.

In step S301, the controller 100 causes the first display device 134 to display a menu screen. FIG. 9 shows an example of a menu screen 810 displayed on the touch screen 133. As shown in FIG. A camera setting icon 811 , a measurement icon 812 , and an end icon 813 are displayed on the touch screen 133 . The user can select a desired function by touching any of these icons.

At step S302, the controller 100 determines whether or not the camera setting function has been selected. For example, when the camera setting icon 811 is touched, it is determined that the camera setting function has been selected. When the camera setting function has not been selected, the process proceeds to step S304. On the other hand, when the camera setting function is selected, the process proceeds to step S303.

In step S303, the controller 100 executes camera setting processing. The camera setting process is a process in which the controller 100 determines shooting conditions and the like for the camera 200 and transmits the shooting conditions and the like to the camera 200 . Details of the camera setting process will be described later. After the camera setting process, the process proceeds to step S304.

At step S304, the controller 100 determines whether the measurement function has been selected. For example, when the measure icon 812 is touched, it is determined that the measure function has been selected. If the measurement function has not been selected, processing proceeds to step S306. On the other hand, when the measurement function is selected, the process proceeds to step S305.

In step S305, the controller 100 executes measurement processing. The measurement process is a process including acquiring an image by the controller 100 from the camera 200 in which shooting conditions and the like are set, and analyzing the state of the sample 500 based on the acquired image. Details of the measurement process will be described later. After the measurement process, the process proceeds to step S306.

In step S306, the controller 100 determines whether or not to end this process. For example, when the end icon 813 is touched, it is determined to end. If not, the process returns to step S301 and repeats the above process. When finished, the process ends.

The camera setting processing performed in step S303 will be described with reference to the flowchart shown in FIG. The camera setting process is a process in which the controller 100 determines shooting conditions and the like for the camera 200 and transmits the shooting conditions and the like to the camera 200 .

In step S401, the controller 100 requests the camera 200 for successive images of the sample 500 in the spinner flask 400. FIG. That is, controller 100 transmits a request signal to camera 200 . In step S402, the controller 100 determines whether reception of images from the camera 200 has started. The controller 100 waits until image reception begins. When image reception has started, the process proceeds to step S403.

At step S<b>403 , the controller 100 receives successive images of the sample 500 transmitted from the camera 200 . Controller 100 analyzes the received image. Based on the analysis result, in step S404, the controller 100 attempts to identify the timing at which the camera 200 should capture the first image and the timing at which the second image should be captured. In step S405, the controller 100 determines whether or not the timing of image acquisition has been specified. If not specified, the process returns to step S403. At this time, acquisition of images from the camera 200 is repeated, and based on the results, determination of the timing of image acquisition is repeatedly attempted. When the timing of image acquisition can be specified, the process proceeds to step S406. More specifically, it is as follows.

Wings 420 of spinner flask 400 rotate about axis of rotation 412 . Therefore, the state suitable for acquiring the first image and the state suitable for acquiring the second image are alternately repeated. Here, the state suitable for obtaining the first image is the state in which the end surface 431 of the blade 420 faces the camera 200 . A state suitable for acquiring the second image is a state in which the major surfaces 432 of the two vanes 420 face the camera 200 .

When the interval at which the camera 200 acquires an image is sufficiently short relative to the rotation speed of the blade 420, photography is performed in all cases in which the end face 431 of the blade 420 faces the camera 200 and in all cases in between. Become. However, it is not necessary to shorten the shooting interval so much. For example, an integer multiple of the period in which the end face 431 of the blade 420 faces the camera 200 should not coincide with the imaging period. In this case as well, the end surface 431 of the blade 420 faces the camera 200 in at least one shot out of a plurality of shots, and the main surfaces 432 of the two blades 420 in at least one shot out of a plurality of shots. faces the camera 200 . The controller 100 analyzes the obtained image and the timing at which the image was obtained, and specifies the timing at which the camera 200 should acquire the first image and the timing at which the second image should be acquired. . Generally, it is sufficient if the first image and the second image are acquired, for example, every few seconds or minutes.

The controller 100 can also instruct the camera 200 on shooting conditions other than the shooting timing. The controller 100 may instruct the camera 200 to shorten the exposure time when the image acquired from the camera 200 is blurred. The controller 100 may instruct the camera 200 to adjust the focus position if the image acquired from the camera 200 is blurred. The controller 100 may command the camera 200 to change the exposure conditions when the image acquired from the camera 200 is too bright or too dark. The exposure condition can be changed by changing the aperture value or the exposure time, or by changing the brightness of the illumination light emitted by the illumination unit 280, or the like.

In step S<b>406 , the controller 100 transmits to the camera 200 an instruction of shooting conditions including the specified timing to acquire the first image and the specified timing to acquire the second image. The camera 200 that has received the instruction sets the imaging conditions of the camera 200 based on the instruction. With this, the camera setting process ends. The process returns to the process of step S304.

The measurement process performed in step S305 will be described with reference to the flowchart shown in FIG. The measurement process is a process in which the controller 100 acquires an image from the camera 200 in which shooting conditions and the like are set, and analyzes the state of the sample 500 based on the acquired image.

In step S<b>501 , the controller 100 requests the camera 200 for information on the shooting conditions currently set for the camera 200 . In response to this request, the camera 200 transmits the current setting conditions to the controller 100 . In step S<b>502 , the controller 100 acquires the setting conditions of the camera 200 transmitted from the camera 200 .

In step S<b>503 , the controller 100 determines whether or not the shooting conditions of the camera 200 are optimized based on the acquired setting conditions of the camera 200 . For example, the acquired setting conditions of the camera 200 may include information as to whether or not optimization has been completed. Information indicating that the optimization has been completed may be recorded in the camera 200 when the shooting conditions are set by the camera setting process described above.

When it is determined in step S503 that the optimization has not been completed, the process proceeds to step S504. In step S504, the controller 100 presents to the user using the first display device 134 that optimization is required. The presentation to the user may be performed by voice or the like instead of display. After presentation, the ongoing measurement process ends.

When it is determined in step S503 that the optimization has been completed, the process proceeds to step S505. In step S505, the controller 100 requests an image from the camera 200. FIG. In response to this request, the camera 200 transmits to the controller 100 an image acquired by shooting based on the set shooting conditions.

In step S<b>506 , controller 100 acquires the image transmitted from camera 200 . This image includes a first image and a second image. The controller 100 performs image analysis based on the acquired image. The imaging conditions of the camera 200 may be used for this analysis. The analysis results include the feature amount of the cell mass, the density of the cells 510, the number of the cells 510, the total number of cells based on the cell density, the distribution of the number of particles for each feature amount of the cell mass, or the pH value of the culture solution 520. and other information may be included. For example, the characteristic amount of a cell aggregate includes the form of the cell aggregate or the size of the cell aggregate, and examples of the form or size of the cell aggregate include the area, volume, average diameter, and maximum diameter of the cell aggregate.

In step S<b>507 , the controller 100 presents the analysis result to the user by displaying it on the first display device 134 or stores it in the first storage circuit 116 . FIG. 12A shows an example of a measurement screen 820 showing analysis results displayed on the touch screen 133 . This measurement screen 820 includes a display 821 indicating that measurement is in progress. The measurement screen 820 also includes an image display area 822 in which an image acquired from the camera 200, particularly a first image for confirming the appearance of individual cell clusters, is displayed. The measurement screen 820 includes a graph 823 showing temporal changes in the turbidity of the culture solution 520 acquired based on the second image, and a graph 824 showing temporal changes in the pH value of the culture solution 520 . Measurement screen 820 includes an end icon 825 for closing measurement screen 820 and ending the measurement process.

Measurement screen 820 may include other information, such as cell mass size. That is, other graphs are displayed on the measurement screen 820 in place of or in addition to the graph 823 showing the time change of the turbidity of the culture solution 520 and the graph 824 showing the time change of the pH value of the culture solution 520. may An example of an effective graph may be one that shows the cell distribution at the measurement timing. For example, in the process of cell culture, the surface area of an enlarged cell mass becomes smaller than the sum of the surface areas of individual cells of the same cell number that exist separately. As a result, as the cell mass becomes larger, the resistance becomes smaller and it becomes difficult to float. That is, as the cell mass becomes larger, the cell mass settles. Under such circumstances, there tends to be a state in which large cell clusters are present at low locations and small cell clusters are floating at high locations. In such a case, it is better to float large cell masses to facilitate the supply of nutrients and oxygen, so the agitation is increased. For example, the rotation speed of blade 420 is increased. The graph displayed on the measurement screen 820 may be, for example, a graph that classifies the size of the cell mass and shows where it is in the culture medium. An example of such a graph may be a graph representing the relationship between the size of the cell mass and the floating position (height). Furthermore, it may be a graph showing the relationship between the size of the cell cluster, the floating position (height), and the number of cell clusters. Examples of such graphs are bubble maps (vertical axis is position, horizontal axis is size, bubble size indicates number of particles), heat map (vertical axis is position, horizontal axis is size, color is (indicating the number of particles), or an image representing the results of statistical processing in terms of color or shape instead of a graph. Figures 12B-12D are examples of such graphs. In Figures 12B to 12D, the size of the cell mass is classified as the morphology of the cells. FIG. 12B shows an example of a three-dimensional bar graph, FIG. 12C shows a bubble map, and FIG. 12D shows a heat map. The controller 100 has a program for creating such graphs.

In this way, the controller 100 can display the first image, the second image, the turbidity of the culture medium 520, in other words, the number of cells in the unit amount of the culture medium 520, the pH value of the culture medium 520, the size of the cell mass, and so on. Furthermore, information such as the morphological characteristics of the cell mass can be obtained.

In step S508, the controller 100 determines whether the analysis result satisfies a predetermined condition. If the predetermined condition is not met, the process proceeds to step S510. On the other hand, when the predetermined condition is satisfied, the process proceeds to step S509. In step S509, the controller 100 performs processing according to the analysis result that satisfies the conditions. This process can be set appropriately.

For example, when the turbidity becomes higher than a predetermined value, when the number of cells per unit of culture fluid reaches a predetermined value, or when the pH value of the culture fluid becomes lower than a predetermined value, the controller 100 , the user is presented with information that the culture medium should be replaced or subculture should be performed. When the controller 100 determines that the shooting conditions of the camera 200 should be changed, it transmits the changed shooting conditions to the camera 200 . The controller 100 notifies the user when a condition preset by the user, such as a condition for terminating culture or a condition for adding a reagent, is met. After the process of step S509, the process proceeds to step S510.

In step S510, the controller 100 determines whether or not to end the measurement process. For example, when the end icon 825 is touched, it is determined to end. If not, the process returns to step S506. At this time, the processing such as the analysis described above is continued. On the other hand, when it is determined in step S510 to end, the process proceeds to step S511.

In step S511, the controller 100 transmits an instruction to the camera 200 to reset the shooting conditions. The camera 200 that has received this instruction resets the set shooting conditions. This process is to prevent the camera 200 from shooting with inappropriate settings when the measurement process is to be performed next. After the process of step S511, the measurement process ends.

Processing of the camera 200 according to this embodiment will be described with reference to the flowchart shown in FIG.

In step S<b>601 , the camera 200 determines whether or not there is an image request from the controller 100 . When there is no image request from the controller 100, the process proceeds to step S604. On the other hand, when there is an image request from the controller 100, the process proceeds to step S602.

In step S602, the camera 200 performs imaging under the currently set imaging conditions. At this time, the camera 200 emits illumination light from the illumination unit 280 to illuminate the sample 500 . The camera 200 transmits the captured image to the controller 100 . In step S<b>603 , the camera 200 determines whether or not there is another instruction from the controller 100 . If there are no other instructions, the process returns to step S602. That is, the camera 200 continues photographing and image transmission. When it is determined in step S603 that there is another instruction, the process proceeds to step S604.

In step S<b>604 , the camera 200 determines whether or not shooting conditions have been received from the controller 100 . If the imaging conditions have not been received, the process proceeds to step S606. On the other hand, when the imaging conditions have been received, the process advances to step S605. In step S<b>605 , the camera 200 sets the shooting conditions received from the controller 100 . After that, the process proceeds to step S606.

In step S<b>606 , the camera 200 determines whether or not there is a request for imaging conditions from the controller 100 . If there is no imaging condition request, the process advances to step S608. On the other hand, if there is a request for imaging conditions, the process advances to step S607. In step S<b>607 , the camera 200 transmits the current set values of the shooting conditions to the controller 100 . After that, the process proceeds to step S608.

In step S608, the camera 200 determines whether or not it has received an instruction from the controller 100 to reset the shooting conditions. If no instruction to reset has been received, the process proceeds to step S610. On the other hand, when the instruction to reset is received, the process proceeds to step S609. In step S609, the camera 200 resets the shooting conditions. The process then proceeds to step S610.

In step S610, the camera 200 determines whether or not to end this process. For example, when it is determined to end, such as when an instruction to turn off the power is input, this process ends. If not, the process returns to step S601 and the above process is repeated.

According to this embodiment, the culture system 10 operates the camera 200 under the control of the controller 100 . Since the controller 100 controls the operation of the camera 200 so that the controller 100 acquires the image it needs, the camera 200 can acquire an optimal image without waste. As a result, the accuracy of analysis by the controller 100 is also improved.

[Third embodiment]
A third embodiment will be described. Here, the points of difference from the second embodiment will be described, and the same reference numerals will be given to the same parts, and the description thereof will be omitted. In the second embodiment, camera 200 is operating under control of controller 100 . In the third embodiment, the controller 100 can control not only the camera 200 but also the stirrer 300 and the support 290 that fixes the camera 200 .

<System configuration>
A schematic configuration example of the culture system 10 according to the present embodiment is shown in the block diagram of FIG. As shown in FIG. 14, configurations of a controller 100 and a camera 200 are similar to those of the second embodiment. The basic configuration of the stirrer 300 is the same as in the case of the second embodiment. The stirrer 300 further includes a third processor 310 , a third memory circuit 316 and a third communication circuit 320 as a third circuit group 305 .

Third processor 310 includes an integrated circuit such as, for example, a CPU, ASIC, or FPGA. The third processor 310 may be configured by one integrated circuit or the like, or may be configured by combining a plurality of integrated circuits or the like. The third processor 310 operates according to a program recorded in the third storage circuit 316 or a recording area within the third processor 310, for example. The third storage circuit 316 includes necessary storage media such as ROM, RAM, and flash memory.

A third communication circuit 320 is a circuit for communicating with the controller 100 . Communication between the controller 100 and the stirrer 300 may be performed by wire or wirelessly. When performed wirelessly, a relatively high-speed communication method such as Wi-Fi may be used, but since the amount of information to be transmitted and received is small, a low-speed but low power consumption communication method such as Bluetooth may be used. .

The third processor 310, the third memory circuit 316, the third communication circuit 320, the third input device 332, the third display device 334, etc. are connected to each other via a third bus line 319. . A motor 354 is also connected to the third bus line 319 via a drive circuit (not shown).

The third processor 310, having received the command from the controller 100 via the third communication circuit 320, controls the rotation speed of the motor 354 based on the command. In this manner, controller 100 can control the rotational speed of blades 420 of spinner flask 400 .

The support 290 according to this embodiment also includes a drive mechanism 294 for changing the position of the fixed camera 200 . Operation of drive mechanism 294 is controlled by fourth processor 292 . Post 290 also includes a fourth communication circuit 291 for communicating with controller 100 . Post 290 can adjust the position of camera 200 under the control of controller 100 .

<Outline of system operation>
An overview of the operation of the culture system 10 according to this embodiment will be described with reference to the flowcharts shown in FIGS. 15A, 15B, and 15C. In this embodiment, the touch screen 133 of the controller 100 can display a menu screen 830 as shown in FIG. That is, the menu screen 830 can include an observation position setting icon 831 , a measurement setting icon 832 , a measurement icon 833 and an end icon 834 . An observation position setting icon 831 is an icon selected when starting to set an observation position. A measurement setting icon 832 is an icon that is selected when starting settings for measurement. A measurement icon 833 is an icon selected when starting measurement. The end icon 834 is an icon selected when instructing end.

In this embodiment, it is assumed that the observation position setting icon 831 is selected and the position of the camera 200 is adjusted. In step S<b>701 , the controller 100 sends a command to the stirrer 300 to rotate the magnet plate 352 . Further, the controller 100 requests images from the camera 200 . In response to these instructions, in step S702, camera 200 starts continuous shooting and starts transmitting the acquired images to controller 100. FIG. Also, in step S703, the stirrer 300 starts rotating the motor 354 to rotate the magnet plate 352. As shown in FIG.

In step S704, the controller 100 analyzes whether or not the position of the camera 200 is appropriate based on the image acquired from the camera 200. FIG. An example of a camera position analysis method will be described with reference to FIGS. 17A and 17B. 17A and 17B, the horizontal axis indicates the elapsed time, and the vertical axis indicates the brightness of the image acquired by the camera 200. FIG. When the camera 200 is properly oriented toward the spinner flask 400 and set at the height at which the sample 500 is placed, the resulting agitation by the blades 420 of the spinner flask 400 is captured as shown in FIG. 17A. The brightness fluctuates periodically. On the other hand, when the installation of the camera 200 is not appropriate, such as when the camera 200 is installed at a position higher than the height of the sample 500, as shown in FIG. variation is not obtained. The controller 100 determines whether the position of the camera 200 is appropriate based on such information, for example.

At step S705, the controller 100 determines whether the camera position is appropriate. If not, the process proceeds to step S706. In step S706, the controller 100 transmits an instruction to change the position of the camera 200 to the post 290 to which the camera 200 is fixed. In step S<b>707 , the post 290 changes the position of the camera based on instructions from the controller 100 . For example, the fourth processor 292 of the stanchion 290 operates the drive mechanism 294 based on the instructions of the controller 100 . Thereafter, the process returns to steps S702 and S703. The camera position is adjusted based on the images captured by the camera 200 as described above.

When it is determined in step S705 that the camera position is appropriate, the process proceeds to step S708. In this embodiment, after setting the camera position, it is assumed that the measurement setting icon 832 is specified on the menu screen 830 shown in FIG. there is

In step S708, the controller 100 requests an image from the camera 200 and requests information on the position of the vanes 420 from the stirrer 300. FIG. The position of vane 420 corresponds to the rotational position of magnetic plate 352 of stirrer 300 . In step S<b>709 , the camera 200 performs continuous shooting and transmits the acquired images to the controller 100 . In step S<b>710 , stirrer 300 transmits information on the rotational position of magnet plate 352 corresponding to the position of blade 420 to controller 100 .

In step S<b>711 , the controller 100 acquires an image from the camera 200 and information on the position of the blades 420 from the stirrer 300 . In step S<b>712 , the controller 100 determines the shooting conditions of the camera 200 and the rotational speed of the blades 420 based on the obtained image and the positional information of the blades 420 . The controller 100 transmits the determined shooting conditions to the camera 200 . The controller 100 also transmits the determined rotation speed of the blades 420 to the stirrer 300 .

The rotation speed of the blades 420 has a range that is generally considered appropriate. Controller 100 identifies the number of revolutions of vane 420 based on the periodic brightness variation of the image as shown in FIG. 17A. Controller 100 determines the number of revolutions so that it falls within an appropriate range. Controller 100 may adjust the number of rotations based on the cell density of sample 500 . For example, when the cell density is high, the rotation speed may be set relatively high, and when the cell density is low, the rotation speed may be set relatively low.

As for the shooting conditions, the controller 100 can determine the exposure conditions based on the brightness of the image and the like. In addition, the controller 100 can determine the image quality, imaging frequency, etc. according to the purpose of the experiment.

In step S<b>713 , the camera 200 sets shooting conditions for the camera 200 based on the information acquired from the controller 100 . In step S<b>714 , stirrer 300 sets the rotation speed of motor 354 based on the information obtained from controller 100 .

In step S715, the controller 100 determines whether the imaging conditions of the camera 200 and the rotation speed of the stirrer 300 are appropriate. If not, the process returns to steps S709 and S710. That is, the above processing is repeated. If determined to be appropriate, the process proceeds to step S716.

Although an example in which the observation position, imaging conditions, and rotation speed are set based on the determination of the controller 100 is shown here, the present invention is not limited to this. The observation position, imaging conditions, and rotation speed may be manually set by the user. For example, a control screen 840 as shown in FIG. 18 may be displayed on the touch screen 133 . On this control screen 840, for example, an observation position setting display 841 is displayed to indicate that the observation position is set. The control screen 840 also includes a cross key 842 . The cross key 842 is a display for inputting an instruction to change the height of the camera 200 or change the orientation of the camera 200, for example. Control screen 840 also includes a live view display 843 of the image captured by camera 200 . The user can grasp the shooting situation while viewing the live view display 843 . Control screen 840 also includes a rotation speed setting icon 844 that is selected when setting the rotation speed of stirrer 300 . When the rotation speed setting icon 844 is selected, the controller 100 transitions to the rotation speed setting screen. The user can, for example, operate the cross key 842 to increase or decrease the rotation speed. Control screen 840 also includes a camera settings icon 845 . When the camera setting icon 845 is selected, the controller 100 transitions to the camera setting screen. The user can set imaging conditions and the like by operating the cross key 842, for example.

Returning to FIG. 15C, the description continues. In step S716, the controller 100 requests an image from the camera 200 and requests information on the position of the vane 420 from the stirrer 300. FIG. In step S<b>717 , the camera 200 performs continuous shooting under the shooting conditions set in step S<b>713 and transmits the acquired images to the controller 100 . In step S718, the stirrer 300 transmits information on the rotational position of the magnet plate 352 rotating under the conditions set in step S714, ie, the position of the blades 420. FIG.

In step S<b>719 , controller 100 acquires an image from camera 200 and information on the position of blade 420 from stirrer 300 . In step S720, the controller 100 performs various analyzes based on the acquired image. The analysis performed here is similar to, for example, the first and second embodiments. In step S<b>721 , the controller 100 presents the image and analysis results to the user using the first display device 134 . Also, the controller 100 records the image and the analysis result in the first memory circuit 116 .

In step S722, the controller 100 determines whether the imaging conditions of the camera 200 and the rotational speed of the stirrer 300 are appropriate. If not, the process returns to step S708. That is, the operation of setting the imaging conditions and the rotation speed in steps S708 to S715 is performed again. When it is determined in step S722 that the settings are appropriate, the process proceeds to step S723. In step S723, the controller 100 determines whether or not to end this process. For example, when an end instruction is input, the process ends. If not, the process returns to steps S717 and S718. On the other hand, when it ends, this process ends.

<Details of system operation>
The operation of the culture system 10 according to this embodiment will be further described. The operation of the controller 100 in the culture system 10 of this embodiment will be described with reference to FIGS. 19 to 21. FIG.

In step S801, the controller 100 causes the touch screen 133 to display the menu screen 830 shown in FIG. 16, for example.

In step S802, the controller 100 determines whether or not the observation position setting icon 831 has been touched to select observation position setting. If the viewing position setting has not been selected, the process proceeds to step S804. On the other hand, when setting the observation position is selected, the process proceeds to step S803. In step S803, the controller 100 executes observation position setting processing. The observation position setting process will be described later. After the observation position setting process, the process proceeds to step S804.

In step S804, the controller 100 determines whether or not the measurement setting icon 832 has been touched to select a setting related to measurement. If no measurement settings have been selected, the process proceeds to step S806. On the other hand, when the measurement setting is selected, the process proceeds to step S805. In step S805, the controller 100 executes measurement setting processing. The measurement setting process will be described later. After the measurement setting process, the process proceeds to step S806.

In step S806, controller 100 determines whether or not measurement icon 833 has been touched to select start of measurement. If no measurements have been selected, processing proceeds to step S808. On the other hand, when measurement is selected, the process proceeds to step S807. In step S807, the controller 100 executes measurement processing. The measurement process will be described later. After the measurement process, the process proceeds to step S808.

In step S808, controller 100 determines whether or not end icon 834 has been touched to select end of processing. If end is not selected, the process returns to step S801. On the other hand, when the end is selected, this process ends.

The observation position setting process performed in step S803 will be described with reference to the flowchart shown in FIG.

In step S901, the controller 100 transmits an instruction to the stirrer 300 to start rotation. In step S902, the controller 100 requests the camera 200 for continuous captured images. In step S903, the controller 100 determines whether reception of continuous images from the camera 200 has started. When reception has not started, processing waits. When reception has started, the process proceeds to step S904.

In step S<b>904 , controller 100 acquires an image from camera 200 . In step S905, the controller 100 analyzes the appropriateness of the position of the camera 200 based on the acquired sequential images, for example, using the method described with reference to FIGS. 17A and 17B. In step S906, controller 100 determines whether the position of camera 200 is appropriate. If the position of camera 200 is not appropriate, processing proceeds to step S907. In step S<b>907 , the controller 100 transmits an instruction to change the position of the camera 200 to the post 290 . As a result, the post 290 moves the drive mechanism 294 to change the position of the camera 200 . After that, the process returns to step S904. The processing of steps S904 to S907 is repeated until the camera 200 is properly positioned.

When it is determined in step S906 that the position of camera 200 is appropriate, the process proceeds to step S908. In step S<b>908 , controller 100 determines that the position of camera 200 is appropriate and notifies strut 290 accordingly. As a result, post 290 fixes the position of camera 200 .

The measurement setting process performed in step S805 will be described with reference to the flowchart shown in FIG.

In step S<b>1001 , controller 100 requests stirrer 300 for the position of blade 420 , that is, the rotational position of magnet plate 352 . In step S1002, the controller 100 requests the camera 200 for an image. In step S<b>1003 , controller 100 determines whether or not it has started receiving information about the position of blade 420 from stirrer 300 and continuous images from camera 200 . When reception has not started, processing waits. When reception has started, the process proceeds to step S1004.

In step S<b>1004 , controller 100 acquires an image from camera 200 . In step S<b>1005 , controller 100 acquires position information of blade 420 . In step S1006, controller 100 analyzes the acquired image.

In step S1007, the controller 100 analyzes the imaging conditions and rotation speed based on the image analysis results. For example, as in the second embodiment, the controller 100 attempts to identify the timing at which the camera 200 should capture the first image and the timing at which the second image should be captured. The controller 100 also sets shooting conditions for the camera 200 . The controller 100 shortens the exposure time when the image acquired from the camera 200 is blurred. The controller 100 adjusts the focus position when the image acquired from the camera 200 is blurred. The controller 100 changes the exposure conditions such as adjusting the aperture value and changing the brightness of the illumination light emitted from the illumination unit 280 when the image acquired from the camera 200 is too bright or too dark. The controller 100 confirms the state of the cells 510 in the culture solution 520 and determines whether or not appropriate stirring is being performed. If the number of cells increases and the cell density appears to be high in the lower portion of the culture solution 520, the rotational speed of the blades 420 is increased.

In step S1008, the controller 100 determines whether or not the setting of the photographing conditions by the camera 200, the rotation speed of the blades 420 by the stirrer 300, and the like are appropriate. When it is determined that the settings are appropriate, this measurement setting process ends. On the other hand, when it is determined that the setting is inappropriate, the process advances to step S1009.

In step S1009, the controller 100 uses the analysis result of step S1007 to determine the photographing conditions of the camera 200, the rotational speed of the blades 420 of the stirrer 300, and the like. In step S1010, the controller 100 sends a rotation speed instruction to the stirrer 300. FIG. In step S<b>1011 , the controller 100 transmits a command regarding shooting conditions to the camera 200 . The stirrer 300 and the camera 200 that have received these commands change their settings. In this way, the measurement conditions for the culture system 10 are appropriately set.

The measurement processing performed in step S807 will be described. The measurement process is generally the same as the measurement process of the second embodiment described with reference to FIG. Different points will be explained.

In the processing performed in steps S508 and S509, the following operations may be performed in addition to the operations described in the second embodiment. For example, when culture of cells 510 progresses and it is determined in step S508 that the rotation speed of blades 420 is not appropriate, controller 100 can determine an appropriate rotation speed and send the command to stirrer 300. For example, when the culture of the cell 510 progresses and it is determined in step S508 that the imaging conditions are not appropriate, the controller 100 can execute the measurement setting process described with reference to FIG.

The camera 200 performs the operation described with reference to FIG. 13 in this embodiment as well as in the second embodiment.

The operation of the strut 290 according to this embodiment will be described with reference to the flowchart shown in FIG.

In step S<b>1101 , the support 290 determines whether or not there is an instruction from the controller 100 to change the position of the camera 200 . If there is no position change instruction, the process advances to step S1103. On the other hand, when there is an instruction to change the position, the process advances to step S1102. In step S<b>1102 , the column 290 operates the drive mechanism 294 based on the instructions from the controller 100 . That is, support 290 changes the position of camera 200 based on instructions from controller 100 . After that, the process proceeds to step S1103.

In step S<b>1103 , the column 290 determines whether or not the controller 100 has issued a position determination instruction. If there is no position determination instruction, the process advances to step S1105. On the other hand, when there is an instruction to fix the position, the process advances to step S1104. In step S<b>1104 , support 290 locks drive mechanism 294 so as to fix the position of camera 200 based on an instruction from controller 100 . After that, the process proceeds to step S1105.

In step S1105, the support 290 determines whether or not to end this process. If not, the process returns to step S1101. That is, the strut 290 repeats the above process. When the process ends, this process ends.

The operation of the stirrer 300 according to this embodiment will be described with reference to the flowchart shown in FIG.

In step S<b>1201 , stirrer 300 determines whether or not there is an instruction from controller 100 to start rotating magnet plate 352 . If there is no instruction to start rotation, the process advances to step S1203. On the other hand, when there is an instruction to start rotation, the process advances to step S1202. In step S1202, the stirrer 300 starts rotating the motor 354. FIG. After that, the process proceeds to step S1203.

In step S<b>1203 , stirrer 300 determines whether controller 100 requests the rotational position of magnet plate 352 . If there is no position request, the process proceeds to step S1205. On the other hand, when there is a position request, the process proceeds to step S1204. In step S<b>1204 , stirrer 300 starts transmitting information on the rotational position of magnet plate 352 to controller 100 . After that, the process proceeds to step S1205.

In step S<b>1205 , the stirrer 300 determines whether or not information regarding the rotation speed has been received from the controller 100 . When the rotational speed has not been received, the process proceeds to step S1207. On the other hand, when the rotation speed is received, the process proceeds to step S1206. In step S<b>1206 , stirrer 300 sets the rotation speed of motor 354 based on an instruction from controller 100 . After that, the process proceeds to step S1207.

In step S1207, the stirrer 300 determines whether or not to end this process. If the process does not end, the process returns to step S1201. On the other hand, when ending, this process ends.

According to this embodiment, controller 100 can control post 290 to change the position of camera 200 . The controller 100 can also control the rotation speed of the magnetic plate 352 of the stirrer 300 to change the rotation speed of the vanes 420 of the spinner flask 400 . Therefore, the controller 100 can finely set the operation of the culture system 10 as a whole.

Each of the above-described embodiments is a matter that has conventionally been adjusted according to the user's sense. By mechanically controlling these matters by the controller 100, detailed control of suspension culture becomes possible. In addition, the culture system 10 can accurately capture the state of floating culture, present it to the user, and record it. These things improve the accuracy of experiments, etc., and expand the possibilities of new experiments that have been difficult so far. In addition, the culture system 10 of the present embodiment enables accurate control of not only laboratory level culture but also mass culture of cells for industrial use. Moreover, the above-described technique can be applied not only to cell culture and the like, but also to bioreactors and the like.

Each embodiment has shown an example in which various analyzes are performed by the controller 100 . However, it is not limited to this. Various analyzes may be performed by a server or the like on the network. Functions such as analysis and control of each device or display of results may be distributed and performed by a plurality of devices.

Among the techniques described in each embodiment, the control mainly described in the flow charts can be implemented using a program. This program can be stored in a storage medium or recording unit. There are various methods of recording in this storage medium or recording unit, and may be recorded when the product is shipped, may be recorded using a distributed storage medium, or may be recorded using download via the Internet. may be

Also, functions similar to the control described above may be realized by artificial intelligence constructed by, for example, deep learning. In other words, it becomes possible to appropriately execute matters that have been judged by humans in the past through deep learning, such as the rotational speed of the blades 420 of the spinner flask 400, the culture density, the pH value of the culture solution, and the like.

In the embodiments, parts described as "parts" (sections or units) may be configured by combining a dedicated circuit or a plurality of general-purpose circuits, and if necessary, according to pre-programmed software It may be configured by combining a microcomputer that operates, a processor such as a CPU, or a sequencer such as an FPGA. Also, it is possible to design such that part or all of the control is taken over by an external device, in which case a wired or wireless communication circuit intervenes. Communication may be performed by Bluetooth communication, Wi-Fi communication, telephone line, or the like, and may be performed by USB or the like. A dedicated circuit, a general-purpose circuit, and a control unit may be integrated into an ASIC.

DESCRIPTION OF SYMBOLS 10... Culture system, 100... Controller, 105... First circuit group, 110... First processor, 116... First memory circuit, 119... First bus line, 120... First communication circuit, 132... First input device 133 Touch screen 134 First display device 136 Input button 200 Camera 202 Camera body 205 Second circuit group 210 Second processor 214 Image processing circuit 216 Second storage circuit 219 Second bus line 220 Second communication circuit 232 Second input device 234 Second display device 250 Lens unit 260 Imaging element 280 Illumination unit 290 Strut 291 Fourth communication circuit 292 Fourth processor 294 Drive mechanism 300 Stirrer 305 Third circuit group 310 Third Processor 316 Third memory circuit 319 Third bus line 320 Third communication circuit 332 Third input device 334 Third display device 352 Magnet plate 354 Motor , 400... spinner flask, 402... container body, 404... lid, 412... rotating shaft, 420... blade, 421... first blade, 422... second blade, 423... third blade, 424... fourth Blade, 431... End face, 432... Main surface, 435... Flat plate, 452... Stirrer bar, 500... Sample, 510... Cell, 511... Monolayer cell, 512... Suspension sample, 520... Culture solution.

Claims (17)

Acquiring a group of images obtained by repeatedly photographing a spinner flask in which cells are cultured in suspension in a culture medium by a camera arranged on the side of the spinner flask,
extracting from the set of images a first image taken when the blades of the spinner flask are along the optical axis of the camera;
An analysis device comprising at least one processor configured to analyze at least one of morphology and size of the cells based on the obtained first image . The set of images further includes a second image taken when the blades of the spinner flask are not aligned with the optical axis of the camera;
The processor
extracting the first image or the second image from the image group;
when extracting the first image, analyzing at least one of the morphology and size of the cell;
When extracting the second image, analyze at least the state of the culture solution
The analysis device according to claim 1. The analysis device according to claim 1 or 2;
a camera for repeatedly photographing the spinner flask to acquire the image group;
including
analysis system. 4. The analysis system according to claim 3 , wherein the processor is configured to control the operation of the camera based on at least one of morphology and size of the cell obtained by the analysis. the set of images further includes a second image taken when the blades of the spinner flask are not aligned with the optical axis of the camera;
The processor transmits conditions for the camera to acquire either the first image or the second image to the camera.
Claim 3 or 4 according to analysis system. Further comprising a drive mechanism for changing the position of the camera,
The processor is configured to control operation of the drive mechanism based on images obtained from the camera.
The analysis system according to any one of claims 3 to 5 . The camera is arranged so that the optical axis of the camera does not face the rotation axis of the blades of the spinner flask,
further comprising a lighting unit that emits illumination light toward the spinner flask from a direction oblique to the optical axis of the camera;
The analysis system according to any one of claims 3 to 6 . The state of the culture medium includes the pH of the culture medium based on the color of the culture medium identified using the second image.
The analysis device according to claim 2. Further comprising a stirrer for rotating the blades of the spinner flask,
The processor is configured to control the rotation speed of the stirrer based on at least one of the morphology and size of the cells obtained by the analysis.
The analysis system according to any one of claims 3 to 7 . The analysis device according to claim 1 or 2 , wherein the processor calculates a distribution for each classification of the feature amount of the cell for each height of the spinner flask. A display unit for displaying the results calculated by the processor,
11. The analysis device according to claim 10 , wherein the display unit displays the relationship between the feature amount of the cells, the total number of cells included in the classification, and the position of the spinner flask in the height direction. The analysis device according to claim 10 or 11 ;
and the camera;
The analysis system, wherein the camera is installed so that the longer aspect ratio is in the height direction. Acquiring a group of images obtained by repeatedly photographing a spinner flask in which cells are cultured in suspension in a culture medium by a camera arranged on the side of the spinner flask;
extracting from the set of images a first image taken when the blades of the spinner flask are along the optical axis of the camera;
analyzing at least one of the morphology and size of the cells based on the obtained first image;
A program that causes a computer to run The processor determines the first image based on at least one of the rotational speed of the blades of the spinner flask, the photographing conditions of the camera, and the relationship between the blades of the spinner flask and the optical axis of the camera specified from the image. extract image
The analysis device according to claim 1. The processor further analyzes the number or density of the cells when extracting either the first image or the second image.
The analysis device according to claim 2. The analysis device according to claim 2, wherein the first image is a monolayer cell image, and the second image is a multilayer cell image. The thickness of the imaging target area of the first image in the optical axis direction of the camera is smaller than the thickness of the imaging target area of the second image in the optical axis direction of the camera.
The analysis device according to claim 2.

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