Patent Application
20200192073
The technology of the disclosure relates to an observation apparatus, an observation method, and an observation program for observing an observation target by relatively moving at least one of a container accommodating the observation target or an imaging unit with respect to the other.
In recent years, various technologies for imaging a subject such as various cells and analyzing an acquired image have been suggested. For example, a method of imaging a pluripotent stem cell such as an embryonic stem (ES) cell and an induced pluripotent stem (iPS) cell, a differentiation-induced cell, or the like using a microscope or the like and determining a differentiation state or the like of the cell by recognizing a feature of the image has been suggested.
The pluripotent stem cell such as the ES cell and the iPS cell has a capability to differentiate into cells of various tissues and has drawn attention for its applicability in regenerative medicine, development of medication, identification of diseases, and the like.
In a case where the cell is imaged as described above, it is preferable to acquire a high magnification wide field-of-view image. In order to do so, for example, JP2016-071117A suggests an imaging method of scanning a range of a cultivation container such as a well plate or a dish by an image forming optical system, capturing an image at a series of predetermined imaging positions, and then, combining a partial image obtained at each imaging position. Consequently, by combining the partial images, the entire cell is imaged in a wide field of view. Such an imaging method is referred to as tiling imaging, and each partial image is referred to as a tile image.
In a case where the image forming optical system relatively scans the cultivation container, a positional deviation occurs due to a change in speed, vibration, or the like, and it is difficult to output accurate partial images.
Therefore, JP2016-125913A discloses a technology for performing re-imaging in a case where a defect occurs in a part of the tile images in the tiling imaging.
However, in the technology disclosed in JP2016-125913A, imaging has to be performed again each time a defect occurs in a part of the tile images. Thus, a processing time for obtaining the final entire image of the cell or the like is increased as the re-imaging is performed.
The technology of the disclosure is conceived in view of the above point, and an object of the technology is to provide an observation apparatus, an observation method, and an observation program capable of acquiring consistent partial images without performing re-imaging.
An observation apparatus according to the technology of the disclosure comprises an imaging unit that images a container accommodating an observation target in an accommodation part and the observation target in a field of view smaller than the accommodation part at predetermined imaging positions and acquires a series of partial images, a movement unit that relatively moves at least one of the container or the imaging unit with respect to the other, a calculation unit that calculates partial shape information representing a partial shape of the container from the partial image obtained by imaging performed by the imaging unit, a storage unit that stores reference shape information representing a shape of the container, and a control unit that corrects the imaging position in a case where the partial image scheduled to be subsequently captured during acquisition of the series of partial images is captured based on the partial shape information and the reference shape information, and causes the series of partial images in which adjacent partial images are consistent to be captured by controlling the movement unit and the imaging unit based on the corrected imaging position.
The predetermined imaging position is not a position of the imaging unit in a focus direction (Z direction) and is a position in a plane (XY plane) in which the container is placed. The partial image is an image that is obtained by imaging by the imaging unit in a field of view of the imaging unit at each of a plurality of predetermined imaging positions. The partial shape is a part that represents a feature of a shape of the accommodation part of the container included in an imaging field of view of the partial image. For example, the partial shape is a shape of a part of an edge of a well in a case where the accommodation part is a well constituting a recessed portion. The partial shape information is shape information representing the partial shape of the container included in the partial image and is information indicating the shape and a position of the accommodation part included in the partial image. The reference shape information of the container is information that is acquired and stored in the storage unit in advance before capturing of the partial image, and is information representing the shape (reference shape) of the container as a reference such as the shape of the accommodation part and the position of the accommodation part in the entire container. Consistency means matching a position of the partial shape of the container in a boundary part between adjacent partial images or reducing a positional deviation in a case where matching cannot be performed.
In the observation apparatus, the calculation unit may calculate reference partial shape information corresponding to an imaging field of view of each partial image from the reference shape information based on the predetermined imaging positions of the series of partial images, and based on the partial shape information in one partial image of the adjacent partial images and the reference partial shape information corresponding to the imaging field of view of the other partial image scheduled to be captured subsequently to the one partial image, the control unit may cause the other partial image to be consistent with the one partial image by correcting the imaging position of the other partial image from the predetermined imaging positions. The reference partial shape information is information that indicates the reference shape (reference partial shape) of the container in each imaging field of view in which the reference shape of the entire container indicated by the reference shape information is divided in units of imaging field of views of the partial images of the imaging unit at each of the predetermined imaging positions.
In the observation apparatus, the control unit may reduce a deviation between a position of a partial shape of the accommodation part included in the one partial image and a position of a partial shape of the accommodation part included in the other partial image in a boundary part between the adjacent partial images by correcting the imaging position of the other partial image from the predetermined imaging positions. The boundary part includes an adjacent side of the adjacent partial images. For example, the boundary part may be a region that extends inwards at a predetermined ratio from the adjacent side, or may be boundary lines of the adjacent partial images.
In the observation apparatus, the storage unit may store shape information representing a shape of an edge of the accommodation part included in the container as the reference shape information, and the calculation unit may calculate shape information representing a shape of an edge of the accommodation part included in the partial image as the partial shape information.
The observation apparatus may further comprise an input unit into which the reference shape information is input, in which the storage unit stores the reference shape information input into the input unit.
The observation apparatus may further comprise a measurement unit that acquires the reference shape information by measuring the shape of the container, in which the storage unit stores the reference shape information obtained by measurement performed by the measurement unit.
An observation method according to the technology of the disclosure comprises an imaging step of imaging a container accommodating an observation target in an accommodation part and the observation target in a field of view smaller than the accommodation part at predetermined imaging positions and acquiring a series of partial images, a calculation step of calculating partial shape information representing a partial shape of the container from the partial image obtained by imaging performed in the imaging step, a storage step of storing reference shape information representing a shape of the container, and a control step of correcting the imaging position in a case where the partial image scheduled to be subsequently captured during acquisition of the series of partial images is captured based on the partial shape information and the reference shape information, and causing the series of partial images in which adjacent partial images are consistent to be captured in the imaging step based on the corrected imaging position.
An observation program according to the technology of the disclosure causes a computer to execute an imaging step of imaging a container accommodating an observation target in an accommodation part and the observation target in a field of view smaller than the accommodation part at predetermined imaging positions and acquiring a series of partial images, a calculation step of calculating partial shape information representing a partial shape of the container from the partial image obtained by imaging performed in the imaging step, a storage step of storing reference shape information representing a shape of the container, and a control step of correcting the imaging position in a case where the partial image scheduled to be subsequently captured during acquisition of the series of partial images is captured based on the partial shape information and the reference shape information, and causing the series of partial images in which adjacent partial images are consistent to be captured in the imaging step based on the corrected imaging position.
Another observation apparatus according to the technology of the disclosure comprises a memory that stores an instruction to be executed by a computer, and a processor configured to execute the stored instruction, in which the processor executes an imaging step of imaging a container accommodating an observation target in an accommodation part and the observation target in a field of view smaller than the accommodation part at predetermined imaging positions and acquiring a series of partial images, a calculation step of calculating partial shape information representing a partial shape of the container from the partial image obtained by imaging performed in the imaging step, a storage step of storing reference shape information representing a shape of the container, and a control step of correcting the imaging position in a case where the partial image scheduled to be subsequently captured during acquisition of the series of partial images is captured based on the partial shape information and the reference shape information, and causing the series of partial images in which adjacent partial images are consistent to be captured in the imaging step based on the corrected imaging position.
According to the technology of the disclosure, the control unit corrects the imaging position in a case where the partial image scheduled to be subsequently captured is captured based on the reference shape information of the container stored in the storage unit and the partial shape information calculated by the calculation unit, and causes the series of partial images in which the adjacent partial images are consistent to be captured based on the corrected imaging position. Since the imaging position of the imaging unit is appropriately corrected during acquisition of the series of partial images and the movement unit and the imaging unit are controlled, re-imaging after the end of a series of imaging or re-imaging of the partial images at the same position by suspending the series of imaging does not occur. Consequently, re-imaging is not necessary, and consistent adjacent partial images can be acquired.
Hereinafter, one example of an embodiment according to the technology of the disclosure will be described with reference to the drawings. The same or equivalent constituents and parts in each drawing will be designated by the same reference signs. Dimensional ratios in the drawings are exaggerated for convenience of description and may be different from the actual ratios.
The observation apparatus is an apparatus for observing an observation target accommodated in a cultivation container 20 placed on a placing stand 10 by a microscope device 30. The placing stand 10 and the microscope device 30 are controlled by a control unit 40. Each configuration will be described in order.
The placing stand 10 is a stage on which the cultivation container 20 can be placed. As illustrated in
A movement unit 12 is attached to the placing stand 10. The movement unit 12 can freely move the placing stand 10 in an X direction and a Y direction that are orthogonal to each other. The X direction and the Y direction are directions orthogonal to a Z direction and are directions orthogonal to each other in a horizontal plane. In the present embodiment, the X direction is set as a main scanning direction, and the Y direction is set as a sub-scanning direction. The movement unit 12 is configured with an actuator that includes a piezoelectric element or the like. Movement of the placing stand 10 in an XY plane is controlled by the control unit 40. By moving the placing stand 10 in the XY plane, the cultivation container 20 on the placing stand 10 moves with respect to the microscope device 30.
In the present embodiment, an example in which a position at which the observation target is observed by the microscope device 30 is changed by moving the placing stand 10 with respect to the microscope device 30 is illustrated. However, the example is not for limitation purposes. The microscope device 30 may be moved with respect to the placing stand 10, or both of the placing stand 10 and the microscope device 30 may be moved. Any aspect can be employed as long as at least one of the cultivation container 20 placed on the placing stand 10 or the microscope device 30 is relatively moved with respect to the other. In the present disclosure, for example, the microscope device 30 is represented as “relatively moving” with respect to the cultivation container 20 even in a case where a position of the microscope device 30 is fixed and only the cultivation container 20 is moving.
Instead of placing the cultivation container 20 on the placing stand 10 and moving the cultivation container 20, the cultivation container 20 may be moved in the X-Y plane by moving a holding unit that holds at least a part of the cultivation container 20.
In the cultivation container 20, a plurality of accommodation parts 22 are formed in a plate 21 having a flat plate shape. For example, a Petri dish, a dish, or a well plate can be used as the cultivation container 20. For example, the accommodation part 22 is a recessed portion having a circular shape in a plan view and is referred to as a well. The accommodation part 22 accommodates the observation target such as various cells immersed in a cultivation liquid. Cells accommodated in the accommodation part 22 include pluripotent stem cells such as an iPS cell and an ES cell, cells of a nerve, skin, cardiac muscle, and a liver that are differentiation-induced from a stem cell, cells of skin, a retina, cardiac muscle, a blood cell, a nerve, and an organ extracted from a human body, and the like.
The microscope device 30 captures a phase difference image of the observation target. In order to obtain a high magnification image, the microscope device 30 captures partial images of the observation target and the cultivation container 20 in a field of view smaller than each accommodation part 22 of the cultivation container 20. As described above, by moving the cultivation container 20 with respect to the microscope device 30, the microscope device 30 scans the cultivation container 20, and a series of partial images is obtained. The partial image is an image that is obtained by imaging by the microscope device 30 in a field of view of the microscope device 30 at each of a plurality of predetermined imaging positions. The predetermined imaging position is not a position of an imaging apparatus in a focus direction (Z direction) and is a position in a plane (XY plane) in which the cultivation container 20 is placed. The position in the focus direction (Z direction) is appropriately adjusted by a focus adjustment mechanism 35 based on a detection result of an autofocus detection unit 38 described later.
The microscope device 30 comprises a light source 31, a slit 32, a condenser lens 33, an objective lens 34, the focus adjustment mechanism 35, an image forming lens 36, an imaging unit 37, and the autofocus detection unit 38.
The light source 31 emits white light. The slit 32 is formed by disposing a ring shaped slit through which the white light is transmitted in a light screen that blocks the white light emitted from the light source 31. Illumination light L having a ring shape is formed by causing the white light to pass through the slit. The condenser lens 33 condenses the illumination light L having the ring shape on the observation target.
The objective lens 34 is arranged to face the condenser lens 33 through the cultivation container 20. The objective lens 34 forms an image of the observation target in the cultivation container 20. The focus adjustment mechanism 35 includes a phase difference lens that can be moved in an optical axis direction (Z direction). By moving the phase difference lens in the optical axis direction, autofocus control is performed, and contrast of the phase difference image captured by the imaging unit 37 is adjusted. The movement of the phase difference lens in the optical axis direction can be implemented by driving an actuator such as a piezoelectric element based on a signal from the control unit 40. However, the piezoelectric element is not for limitation purposes, and the phase difference lens can be driven using other known configurations as long as the phase difference lens can be moved in the Z direction. In addition, a magnification of the phase difference lens may be configured to be changeable. Specifically, a phase difference lens or the focus adjustment mechanism 35 having a different magnification may be configured to be replaceable. The replacement may be automatically performed or may be manually performed by a user.
The phase difference image that passes through the focus adjustment mechanism 35 is incident on the image forming lens 36, and the image forming lens 36 forms the phase difference image on the imaging unit 37.
The imaging unit 37 captures the phase difference image formed by the image forming lens 36. The imaging unit 37 is an imaging element such as a charge-coupled device (CCD) image sensor or a complementary metal-oxide semiconductor (CMOS) image sensor. As the imaging element, an imaging element in which color filters of red, green, blue (RGB) are disposed may be used, or a monochrome imaging element may be used.
Hereinafter, the objective lens 34, the focus adjustment mechanism 35, the image forming lens 36, and the imaging unit 37 will be collectively referred to as an image forming optical system C.
The autofocus detection unit 38 detects a Z-directional position of the cultivation container 20 installed on the placing stand 10. Specifically, the autofocus detection unit 38 comprises a first displacement sensor 38a and a second displacement sensor 38b. The first displacement sensor 38a and the second displacement sensor 38b are arranged in the X direction illustrated in
Z-directional positional information of the cultivation container 20 detected by the autofocus detection unit 38 is output to the control unit 40. The control unit 40 performs the autofocus control by controlling the focus adjustment mechanism 35 based on the input positional information. The detection of the position of the cultivation container 20 by the first displacement sensor 38a and the second displacement sensor 38b, and the autofocus control will be described in detail later.
Next, a configuration of the control unit 40 controlling the microscope device 30 will be described.
The control unit 40 controls the entire microscope device 30 as described above and executes various processes. The control unit 40 includes a microscope device control unit 41, a scanning control unit 42, a display control unit 43, a calculation unit 44, a storage unit 45, an input unit 46, and a display unit 47. The control unit 40 is configured with a computer that comprises a central processing unit (CPU), a semiconductor memory, and the like. In the control unit 40, an observation program according to one embodiment of the present invention is installed in the storage unit 45. The microscope device control unit 41, the scanning control unit 42, and the display control unit 43 illustrated in
The microscope device control unit 41 controls the focus adjustment mechanism 35 based on the Z-directional positional information of the cultivation container 20 detected by the autofocus detection unit 38 as described above. By driving the focus adjustment mechanism 35, the phase difference lens moves in the optical axis direction, and the autofocus control is performed.
In addition, the microscope device control unit 41 controls imaging performed by the imaging unit 37 in a case where the cultivation container 20 is scanned. Specifically, a timing of imaging during scanning is stored in advance in the storage unit 45. The microscope device control unit 41 performs imaging in accordance with the timing.
The scanning control unit 42 controls driving of the movement unit 12 and moves the placing stand 10 in the X direction and the Y direction.
The display control unit 43 generates one composite image by combining the series of partial images captured by the microscope device 30 and displays the composite image on the display unit 47.
The calculation unit 44 calculates partial shape information that is shape information of a partial shape of the cultivation container 20 from the partial images of the cultivation container 20 and the observation target obtained by imaging performed by the imaging unit 37. The partial shape is a part that represents a feature of a shape of the accommodation part 22 of the cultivation container 20 included in an imaging field of view of the partial image. In the present embodiment, the partial shape is a part that represents a feature of the shape and a position of the accommodation part 22 included in the imaging field of view of each partial image. The partial shape information is information that indicates the shape or the position of the accommodation part 22 included in the partial image. In the present embodiment, particularly, the calculation unit 44 calculates the partial shape information from a shape of an edge of the accommodation part 22 of the cultivation container 20 included in the partial image.
The storage unit 45 stores the observation program that implements each function unit. In addition, the storage unit 45 stores, in advance, reference shape information that is obtained by measuring a shape of the cultivation container 20 in advance. The reference shape information is information that is acquired in advance before capturing the partial images, and is obtained by measuring the cultivation container 20 not accommodating the observation target in the accommodation part 22 by a measurement unit such as a laser length measurement device. For example, the reference shape information is information that represents the shape of the accommodation part 22 and the position of the accommodation part 22 in the entire cultivation container 20. In the present embodiment, the reference shape information is the shape of the edge of the accommodation part 22 of the cultivation container 20. The measurement unit used for measurement is preferably a device that has different accuracy from the microscope device 30. Particularly, the measurement unit is preferably a device that can measure the shape of the edge of the accommodation part 22 with higher accuracy than the microscope device 30. The shape of the cultivation container 20 may be measured in advance using the microscope device 30 as the measurement unit.
The reference shape information of the cultivation container 20 stored in the storage unit 45 includes reference partial shape information. The reference partial shape information is information that indicates a reference shape (reference partial shape) of the cultivation container 20 in each imaging field of view in which a reference shape of the entire cultivation container 20 indicated by the reference shape information is divided in units of imaging field of views of the imaging unit 37 at the predetermined imaging positions. Specifically, assuming that the imaging unit 37 reaches the series of imaging positions by scanning, the shape of the edge of the accommodation part 22 of the cultivation container 20 that enters each imaging field of view of the imaging unit 37 is the reference partial shape, and shape information of the reference partial shape is the reference partial shape information. A series of reference partial shape information is obtained in correspondence with all imaging positions. The reference partial shape information may not be stored in the storage unit 45 and may be calculated by the control unit 40 as necessary based on the reference shape information.
The storage unit 45 stores the series of imaging positions of the imaging unit 37 as relative coordinate positions between the placing stand 10 and the microscope device 30.
The input unit 46 comprises a mouse, a keyboard, and the like and receives various necessary data and various setting inputs from the user. For example, the input unit 46 of the present embodiment receives an input of information related to container information of the cultivation container 20, the reference shape information, the reference partial shape information, and the imaging positions. The imaging positions may be determined in advance by the control unit 40 based on the reference shape information input into the input unit 46.
The display unit 47 comprises, for example, a liquid crystal display and displays a composite phase difference image generated by the display control unit 43 as described above. The display unit 47 may be configured with a touch panel and double as the input unit 46.
Next, movement control of the placing stand 10 by the scanning control unit 42 and control of the microscope device 30 by the microscope device control unit 41 will be described in detail.
In the present embodiment, the placing stand 10 is moved in the X direction and the Y direction under control of the scanning control unit 42, and the microscope device 30 two-dimensionally scans the inside of the cultivation container 20. During the scanning, partial images of the cultivation container 20 and the observation target are captured in each field of view of the microscope device 30. In the present embodiment, a well plate that includes six accommodation parts 22 is used as the cultivation container 20.
As illustrated in
In the present embodiment, as illustrated in
In a case where the field of view R is moving in an arrow direction (a leftward direction in
The detection of the cultivation container 20 using the first displacement sensor 38a and the detection of the cultivation container 20 using the second displacement sensor 38b are switched depending on the movement direction of the field of view R. Accordingly, the Z-directional positional information of the cultivation container 20 at the position of the field of view R can be always acquired before the capturing of the phase difference images in the field of view R.
Based on the Z-directional positional information of the cultivation container 20 detected beforehand as described above, the microscope device control unit 41 performs the autofocus control by controlling driving of the focus adjustment mechanism 35. Specifically, a relationship between the Z-directional positional information of the cultivation container 20 and a movement amount of the image forming optical system C in the optical axis direction is set in advance in the microscope device control unit 41. The microscope device control unit 41 obtains the movement amount of the image forming optical system C in the optical axis direction based on the input Z-directional positional information of the cultivation container 20 and outputs a control signal corresponding to the movement amount to the focus adjustment mechanism 35. The focus adjustment mechanism 35 is driven based on the input control signal. Accordingly, the phase difference lens moves in the optical axis direction, and focus adjustment corresponding to the Z-directional position of the cultivation container 20 is performed.
In a case where scanning is performed as described thus far, for example, one accommodation part 22 of the cultivation container 20 is covered with nine partial images P divided by dot-dashed lines as illustrated in
First, the control unit 40 receives an input for the container information related to the cultivation container 20 in the input unit 46 (step S101). For example, the container information of the cultivation container 20 includes specifications (a size, a number, intervals, and the like of accommodation parts 22) of the cultivation container and a model number and a brand of the cultivation container. Information of the number of accommodation parts 22 of the currently used cultivation container 20, the intervals of the accommodation parts 22, a diameter of the accommodation part 22, and the like is obtained from the container information. In addition, since a position and the like of arrangement of the accommodation parts 22 may vary depending on the brand even in a case where the specifications are the same, information of the brand is also effective for specifying the shape of the cultivation container 20.
Next, the control unit 40 measures the shape of the cultivation container 20 not accommodating the observation target in the accommodation part 22 by the measurement unit (step S102). As described above, the measurement unit may not be included in the configuration of the observation apparatus and may be an external device or the microscope device 30. In a case where the shape of the cultivation container 20 is measured, the container information acquired in step S101 is used. For example, the cultivation container 20 is irradiated with laser light by the laser length measurement device, and a solid shape of the container is measured using reflected laser light. At this point, a measurement result of the laser length measurement device can be corrected using the container information. Accordingly, the measurement result of the solid shape of the cultivation container 20 can be more accurate.
The control unit 40 stores shape information that represents the shape of the cultivation container 20 and is obtained in step S102 in the storage unit 45 as the reference shape information (step S103). Step S101 to step S103 are preprocesses for observation performed by the observation apparatus. Accordingly, step S101 to step S103 may be executed in advance regardless of step S104 and subsequent steps.
The control unit 40 starts scanning by starting imaging performed by the microscope device 30 while moving the placing stand 10 by the scanning control unit 42 (step S104).
The control unit 40 captures the partial images of the cultivation container 20 and the observation target by scanning of the microscope device 30 (step S105). Generally, the partial images are captured in a case where the image forming optical system C of the microscope device 30 reaches the predetermined imaging positions. However, the adjacent partial images may be inconsistent due to vibration, acceleration and deceleration, or the like. In such a case, the control unit 40 changes the imaging position of the subsequently captured partial image by controlling the imaging unit 37 and the movement unit 12. Details of a process of capturing the partial images will be described later.
The control unit 40 as a calculation unit calculates the partial shape information representing the partial shape of the cultivation container 20 from the partial images captured in step S105 (step S106). In the present embodiment, the calculated partial shape information is information that indicates the edge of the circular shape of the accommodation part 22 of the cultivation container 20. There is a high likelihood that a part of the edge of the circular shape is included in the partial image. The shape of the edge is effective for checking the consistency between the adjacent partial images. For example, as illustrated in
The control unit 40 stores the calculated partial shape information in the storage unit 45 (step S107).
The control unit 40 decides whether or not the scanning is completed by determining whether or not the imaging is finished at all imaging positions by the microscope device 30 (step S108). In a case where the scanning is not completed (step S108: NO), the process from step S105 is repeated. In a case where the scanning is completed (step S108: YES), the control unit 40 finishes the process of the observation method.
Next, the process of capturing the partial images in step S105 will be described in detail.
The control unit 40 specifies the subsequent imaging position (step S201). The imaging positions are stored in advance in the storage unit 45 and are sequentially specified along with a progress of the scanning.
The control unit 40 decides whether or not the subsequent imaging position specified in step S201 is the initial imaging position in the series of imaging positions (step S202). In a case where the subsequent imaging position is the initial imaging position (step S202: YES), the partial images are captured at the predetermined imaging positions (step S203), and a return is made to the flowchart in
The control unit 40 reads out the reference partial shape information of a position corresponding to the subsequent imaging position from the storage unit 45 (step S204). Alternatively, the control unit 40 may calculate the reference partial shape information corresponding to the subsequent imaging position from the reference shape information based on the reference shape information stored in the storage unit 45 and the subsequent imaging position.
The control unit 40 reads out the partial shape information of the captured partial image adjacent to the subsequent imaging position specified in step S201 from the storage unit 45 (step S205). Accordingly, the partial shape of the captured partial image and the reference partial shape information corresponding to the imaging position that is adjacent to the partial shape and is subsequently imaged are read out.
Based on the read partial shape information and reference partial shape information, the control unit 40 calculates a positional deviation between the imaging position of the captured partial image and the subsequent imaging position and corrects the subsequent imaging position (step S206). Specifically, in a boundary part between the captured partial image and the imaging field of view of the subsequent predetermined imaging position, the control unit 40 calculates a degree to which the position of the edge of the accommodation part 22 deviates in the imaging field of view of the subsequent imaging position from the position of the edge of the accommodation part 22 in the captured partial image. The boundary part includes an adjacent side of the adjacent partial images or an adjacent side between the partial image and the imaging field of view at the subsequent imaging position. For example, the boundary part may be a region that extends inwards at a predetermined ratio from the adjacent side, or may be boundary lines of the adjacent partial images. The control unit 40 decides a direction in which correction for shifting the subsequent imaging position is to be performed such that the position of the partial shape of the captured partial image is arranged in consistency with the position of the reference partial shape in the imaging field of view of the subsequent imaging position in the boundary part. It is preferable that the control unit 40 corrects the imaging position to completely remove the deviation. However, in a case where correcting the imaging position to completely remove the deviation needs to stop or excessively drive the movement unit 12 or causes an unreasonable imaging timing of the imaging unit 37, it may not be necessary to completely remove the deviation. In this case, the control unit 40 corrects the imaging position in a direction in which the deviation is reduced as far as possible.
The control unit 40 images the cultivation container 20 and the observation target at the imaging position corrected in step S206 by controlling the movement unit 12 and the imaging unit 37 (step S207). That is, at least one of the scanning direction or the timing of the imaging performed by the imaging unit 37 is adjusted such that the imaging unit 37 can perform the imaging at the corrected imaging position. In a case where the reference partial shape of the subsequent imaging position does not deviate from the partial shape of the partial image, the control unit 40 does not adjust the scanning direction and the timing of the imaging.
A specific example will be described with reference to
In the example illustrated in
Since capturing is performed up to the partial image P2, the reference partial shape information that indicates a reference partial shape BS included in the partial image B1 corresponding to the imaging position subsequent to the partial image P2 is read out in step S204. Then, in step S205, the partial shape information that indicates a partial shape PS included in the partial image P2 adjacent to the partial image B1 is read out. As illustrated in
As illustrated in
In a case where the shape of the cultivation container 20 is measured in advance, the container information is input in advance. Accordingly, the cultivation container 20 can be measured using the container information, and the shape of the cultivation container 20 is obtained with higher accuracy. However, the container information may not be necessarily input. Even in a case where the container information is not present, the cultivation container 20 can be measured in advance based on various inputs provided by the user or information of the previously used imaging positions.
While one embodiment of the present disclosure is described thus far, the technical idea of the present disclosure is not limited to the embodiment. Various modifications can be made.
In the embodiment, the reference partial shape information corresponding to the series of imaging positions is calculated from the reference shape including the shape of the entire cultivation container 20 and is compared with the partial shape information indicating the captured adjacent partial shape as the imaging position. However, instead of comparing the adjacent reference partial shape information and partial shape information as the imaging position, the reference partial shape information and the partial shape information corresponding to the same location as the imaging position may be compared. Even in this case, a deviation amount of the position of the reference partial shape from the position of the partial shape can be determined, and the subsequent imaging position can be corrected in accordance with the deviation amount. In addition, the reference shape information and the partial shape information can be directly compared without using the reference partial shape information. In this case, for example, a position to which the partial shape corresponds in the reference shape indicated by the reference shape information is specified, and a positional deviation between the partial shape and the reference shape can be determined by comparing the specified position with the imaging position at which the partial shape is actually imaged. The subsequent imaging position can be corrected based on the determined deviation.
In the embodiment, the partial shape information and the reference partial shape information (reference shape information) are calculated using the edge of the accommodation part 22 of the cultivation container 20 as a feature. However, instead of the edge of the accommodation part 22, an edge of the entire cultivation container 20 can be calculated as a feature. Alternatively, a mark having a specific shape such as a protruding portion or a recessed portion can be disposed in the cultivation container 20, and a shape of the mark can be calculated as a feature.
In the embodiment, the imaging position is corrected based on the comparison between the reference partial shape and the partial shape. However, considering one accommodation part 22 illustrated in
An observation process that is executed by causing the CPU to read software (program) in the embodiment may be executed by various processors other than the CPU. In this case, the processors are illustrated by a programmable logic device (PLD) such as a field-programmable gate array (FPGA) of which a circuit configuration can be changed after manufacturing, a dedicated electric circuit such as an application specific integrated circuit (ASIC) that is a processor having a circuit configuration dedicatedly designed to execute a specific process, and the like. In addition, the observation process may be executed by one of the various processors or may be executed by a combination of two or more processors of the same type or different types (for example, a plurality of FPGAs and a combination of a CPU and an FPGA). In addition, a hardware structure of the various processors is more specifically an electric circuit in which circuit elements such as semiconductor elements are combined.
In the embodiment, an aspect in which the program of the observation process is stored (installed) in advance in the storage unit 45 is described. However, the aspect is not for limitation purposes. The program may be provided in a form of a recording on a recording medium such as a compact disk read only memory (CD-ROM), a digital versatile disk read only memory (DVD-ROM), and a Universal Serial Bus (USB) memory. In addition, the program may be downloaded from an external device through a network.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2017-186414 | Sep 2017 | JP | national |
This application is a Continuation of PCT International Application No. PCT/JP2018/031777 filed on Aug. 28, 2018, which claims priority under 35 U.S.C § 119(a) to Japanese Patent Application No. 2017-186414 filed on Sep. 27, 2017. Each of the above application(s) is hereby expressly incorporated by reference, in its entirety, into the present application.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/JP2018/031777 | Aug 2018 | US |
| Child | 16798624 | US |
