Dish, embryo culture apparatus using the dish, and method for imaging an embryo

Abstract

To provide a high-reliability optical system for imaging a treated egg.SOLUTION: A plurality of dishes used in an embryo culturing device that cultures an embryo is formed of transparent resin or glass, and a plurality of wells being recesses each having a diameter capable of housing a treated egg is formed on a bottom part of each of the dishes. On at least one of a bottom part side and a side part side of the well, optical function parts having respective predetermined optical functions, which is a lens, a lens array, a light guiding path, a diffusion plate and the like are formed. Since at least some of the optical function parts are provided on the dish side, the embryo culturing device can be made compact, and its reliability can be enhanced.SELECTED DRAWING: Figure 5

Description

Technical Field

[0001] The present disclosure relates to technologies such as a dish for culturing embryos and an embryo culturing apparatus using this dish.

Background Art

[0002] Social demands for infertility treatment are increasing, and various apparatuses for culturing treated eggs subjected to fertilization procedures under a microscope or the like have been proposed and sold. In recent years, apparatuses that are provided with a camera inside such culturing apparatuses, image the treated eggs during culturing, and display the captured images on an external display as necessary have also become widespread (for example, Patent Document 1 below). In such embryo culturing apparatuses, the treated eggs are placed in a tray (hereinafter also referred to as a dish) that stably holds them, and the entire tray is conveyed into the field of view of the camera at a predetermined interval and imaged. Since an embryologist can determine the possibility of fertilization by observing this image, it is not necessary to take out the treated eggs outside the culturing apparatus only for observation.

[0003] Although the treated eggs are approximately spherical with a diameter of about 100 to 200 μm, since the camera that images them only images them from one direction, the captured images become planar. For this reason, attempts have also been made to control the lens position of the camera to change the focal position and extract three-dimensional information from the treated eggs. In addition, the illuminance and angle of the light applied to the treated eggs are adjusted to capture favorable images.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] However, if an attempt is made to change the focal position or the like, the optical system of the camera becomes complicated, and it is not easy to control for accurately changing the focal position in micron units. Furthermore, adjusting the illuminance and angle of illumination also complicates the configuration of the illumination system. If various such functions are incorporated into the illumination device and camera for imaging, the number of movable parts increases, the device configuration becomes complicated, and it is likely to cause malfunctions. Since the embryo culture device handles the eggs of patients as part of infertility treatment, malfunctions of the device are likely to lead to the failure of infertility treatment. For this reason, a highly reliable optical system for imaging the treated eggs is required.

Means for Solving the Problems

[0006] One aspect of the present disclosure is an aspect as a dish used in an embryo culture device for culturing embryos. This dish is formed of a transparent resin or glass, and a plurality of wells, which are depressions having a diameter capable of accommodating treated eggs, are formed at the bottom of the dish, and an optical functional part having a predetermined optical function is formed on at least one of the bottom side and the side part of the well. By doing so, since the optical functional part is formed on a part of the dish, a part of the optical device on the embryo culture device side can be omitted, and the embryo culture device can be configured compactly. In addition, since the optical functional part is provided on the side of the dish, when the optical functional part is soiled or the like, it is only necessary to replace the dish, and the need to replace or repair the optical device on the embryo culture device side is suppressed.

Brief Description of the Drawings

[0007]

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Mode for Carrying Out the Invention

[0008] A. First Embodiment: (A-1) Hardware Configuration: FIG. 1 is a plan view of an embryo culture device (also referred to as an incubator) 10, and FIG. 2 is an explanatory drawing schematically showing the inside of one culture chamber of the embryo culture device 10. This embryo culture device 10 is for culturing an egg that has undergone in vitro fertilization treatment (hereinafter referred to as a treated egg) for a certain period in a predetermined culture environment, that is, at a constant temperature and humidity. Since the treated egg is not necessarily a fertilized egg, it is cultured in the embryo culture device 10 for a predetermined period. Note that the in vitro fertilization treatment may be intracytoplasmic sperm injection performed under a microscope, or may be normal in vitro fertilization performed by combining an egg and sperm in a predetermined container. The fertilization method of the treated egg to be cultured is not limited.

[0009] As shown in Fig. 1, this embryo culture device 10 is equipped with a total of nine culture units for embryo culture, with five culture units 11 - 15 arranged in the lower row and four culture units 21 - 24 arranged in the upper row. Hereinafter, when collectively referring to the members related to the culture units, they are denoted as culture units 11 - 24. Of course, the number of culture units is not limited. Each of the culture units 11 - 24 includes a culture chamber R11 - R24, a lid portion 17 that seals each of the culture chambers R11 - R24, and switches SW11 - SW24. By operating the switches SW11 - SW24, the lid portions 17 of the corresponding culture units 11 - 24 are opened and closed by a drive mechanism (not shown). Of course, the culture units 11 - 24 may have a structure that can be manually opened and closed. A silicon rubber seal portion is provided on the culture chamber R11 - R24 side of the lid portion 17, and when the lid portion 17 is closed, the inside of the culture chambers R11 - R24 is kept airtight by this seal portion.

[0010] Carbon dioxide gas (CO2) and nitrogen gas (N2) are supplied to the embryo culture device 10 from an external gas cylinder. Gas ports 18, 19 through which these gases are supplied are provided on the back surface of the embryo culture device 10. Also, filter ports 28, 29 to which a filter 27 is attached are provided on the back surface of the embryo culture device 10. This filter 27 is for removing foreign matters such as dust in the outside air taken in by a built-in pump (not shown). Inside the embryo culture device 10, the carbon dioxide gas CO2, nitrogen gas N2 input from the gas ports 18, 19, and the air input through the filter 27 are mixed at a predetermined ratio to generate a mixed gas. The ratio of each gas in the mixed gas is measured by a sensor (not shown) and kept at a constant ratio at all times.

[0011] The case body 20 is provided with a display 70 on its surface. The surface of the display 70 is a touch panel, and various information can be displayed by tapping buttons or the like shown on it. Further, the case body 20 is provided with a general-purpose connector 26 such as a USB-C or Thunderbolt (registered trademark), and various information can also be input by connecting an input device such as a keyboard or a pointing device such as a mouse here. The display example on the display 70 and the case of connecting other devices to the general-purpose connector 26 will be described later.

[0012] Figure 2 shows a state in which the lid portion 17 is opened in the culture unit 11. Culture chambers R11 to R24 are prepared in each of the culture units 11 to 24. Since the structures of the culture chambers R11 to R24 are the same, hereinafter, the culture chamber R11 will be taken as an example for description. The culture chamber R11 is provided with a holding frame 180 on which a dish 191 is placed. A supply port 43 and an exhaust port 44 are provided on the bottom surfaces of the culture chambers R11 to R24. The supply port 43 is an opening for supplying the mixed gas to the culture chamber R11. Further, the exhaust port 44 is an opening for refluxing the mixed gas. The exhaust port 44 is provided at a position covered by the holding frame 180. As described above, when the lid portion 17 is closed, the culture chamber R11 is made substantially airtight, so the internal gas environment is kept constant. However, in order to make the temperature distribution in the culture chamber R11 uniform, the mixed gas is supplied from the supply port 43 and exhausted from the exhaust port 44.

[0013] In the culture chamber R11, panel heaters (not shown) are provided on its side walls and bottom surface. The inside of the culture chamber R11 is maintained at a constant temperature by the panel heaters. Although the description is omitted, in addition to the heaters, temperature sensors, gas concentration sensors, and humidity sensors (not shown) are provided in the culture chambers R11 to R24, and the temperature, gas concentration, and humidity inside the culture chamber R11 can be detected. By feeding back the signals from these sensors, as a result, the inside of the culture chamber R11 is maintained at a constant temperature and humidity, and its gas concentration is also kept constant. Instead of detecting the temperature, etc. inside the culture chamber R11, it is also possible to keep the environment inside the culture chamber R11 constant by keeping the temperature, humidity, gas concentration, etc. of the air-fuel mixture supplied from the supply port 43 constant.

[0014] The holding frame 180 of the culture chamber R11 has a dish 191 containing the treated eggs placed therein. The dish 191 shown in FIG. 2 has 25 microwells 195 for accommodating the treated eggs, arranged in a 5×5 pattern, at the bottom 196. Such depressions for accommodating the treated eggs are simply called wells, but in this embodiment, since 25 wells are densely provided in a particularly small area and the depressions themselves are formed smaller than conventional wells, they are called microwells. The form of the dish 191 is shown in FIG. 3. This dish 191 is accommodated in the accommodation location of the culture chamber R11 with the treated eggs accommodated in the microwells 195. The dish 191 has a cylindrical shape with a diameter of about 35 mm for easy handling, and an outer peripheral wall 194 with a height of about 10 mm stands upright from the bottom 196 on the outer periphery. Further, a circular partition wall 192 with a thickness of about 2 mm is provided inside so as to surround the microwells 195. The inside of this partition wall 192 is filled with a culture solution. The treated eggs are accommodated in the microwells 195 filled with the culture solution. The 25 microwells 195 surrounded by the partition wall 192 and filled with the culture solution are further covered with mineral oil.

[0015] By doing so, when handling the dish 191, it is easy for the embryo culturist to hold, and since it is only necessary to fill the culture solution in a small compartment partitioned by the partition wall 192, the culture solution is not wasted. Also, since the 25 microwells 195 for accommodating the treated eggs are within a few millimeters square, they can be easily imaged by one camera unit for the use described later. If the imaging area is narrow, the resolution of the image per treated egg can be increased, and the accuracy of judgment when making a judgment such as fertilization using this image can be increased. Further, the image of the imaged microwell 195 is displayed on the display 70 which is a display unit provided at the upper right end of the upper stage of the upper surface of the case body 20 as needed.

[0016] The dish 191 shown in FIG. 3 has a recess 197 formed at one location outside the outer peripheral wall 194. This recess 197 is used to fit into a protrusion 182 provided on a holding frame 180 that holds the dish 191 when the circular dish 191 is accommodated in the culture chamber R11, and is used to position the dish 191 at a predetermined position. The holding frame 180 is made in a shape that can accommodate about half the circumference of the dish 191, and the dish 191 is stably held at a predetermined position within the culture chamber R11. Note that the dish 191 is not limited to a circular shape, and may be other shapes such as a rectangle, an ellipse, or a trapezoid. Also, the partition wall 192 is not limited to a circular shape, and may be other shapes such as a square. Further, the partition wall 192 may not be provided, and the entire dish 191 may be filled with the culture solution.

[0017] FIG. 4 is a cross-sectional view taken along line IV-IV in FIG. 2. For clarity, the hatching of the cross-sections of the lid portion 17 and the dish 191 is omitted, and the camera module 51 and the lens module 52 are shown as external views. Reference numeral H11 indicates a continuous side wall and bottom forming the culture chamber R11. As shown in the drawing, directly above the dish 191, an illumination unit 170 provided inside the lid portion 17 is arranged. Further, an opening 198 is provided directly below the microwell 195 of the dish 191 at the bottom H11 of the culture chamber R11, and the camera module 51 and the lens module 52 attached to the case body 20 are arranged. The microwell 195 of the dish 191 is illuminated by the illumination unit 170 provided in the lid portion 17, and the treated eggs 25 accommodated in the microwell 195 are imaged by the camera module 51.

[0018] An optical function unit 200 is formed at the bottom position corresponding to the microwell 195 of the dish 191. The optical function unit 200 in the first embodiment is shown in FIG. 5. FIG. 5 is an enlarged view of part V in FIG. 4. In the example shown in FIG. 5, the optical function unit 200 is a lens array LA of convex lenses provided at a position corresponding to the microwell 195. The lens array LA is composed of 5×5 convex lenses, similar to the 5×5 microwells 195. Since the optical function unit 200 is formed on the bottom 196 of the dish 191, its position exactly corresponds to the position of the microwell 195 formed on the same bottom 196.

[0019] This lens array LA is integrated with the lens of the lens module 52 to form an accurate image on the imaging element 53 of the camera module 51. Specifically, the light incident as parallel light by the lens groups L1, L2, L3 of the lens module 52 is imaged on the imaging element 53. When parallel light enters from the outside, the focal length of the lens array LA is adjusted so as to be focused on the treated eggs 25 in each microwell 195. Therefore, the images of the treated eggs 25 in each microwell 195 are focused on the imaging element 53.

[0020] When the optical system is formed in this way, a convex lens of the lens array LA is arranged one by one directly below each microwell 195, and an enlarged image of each microwell 195 can be formed on the imaging element 53. The imaging element 53 is not in focus except for the images formed by the respective lenses of the lens array LA. That is, while imaging the existence regions of the 5×5 microwells 195 collectively, the portions of each microwell 195 are enlarged, so that the resolution of each microwell 195 on the imaging element 53 is enhanced as compared with the case of imaging without the lens array LA.

[0021] In addition, the apparent angle of view with respect to each microwell 195 becomes smaller, and both the microwell 195 located at the center CX of the dish 191 and the microwell 195 located at the periphery can be imaged in a state of being viewed almost directly from below. Therefore, distortion and the like of the image of each microwell 195 in the image obtained by imaging 5×5 microwells 195 at a time can be reduced.

[0022] FIG. 6 is an explanatory diagram illustrating the relationship between such image distortion and correction. In order to image a plurality of microwells 195 at a time, it is impossible to place all the microwells 195 directly below the optical axis of the camera module 51. For this reason, when a dish without the lens array LA as the optical function unit 200 is used, distortion occurs in the image obtained by imaging each microwell 195. In the case of a color image, there are also cases where blurring or coloring due to aberration occurs. Focusing on the image of one microwell 195 and the treatment egg 25 accommodated therein, since it is not directly below the optical axis, the contour of the microwell 195 formed in a perfect circle is distorted as shown by the dashed line IM in the figure and does not become a perfect circle. Naturally, the same distortion also occurs in the photographed image of the treatment egg 25. On the other hand, when imaging is performed using the dish 191 provided with the lens array LA as the optical function unit 200 at the bottom 196, the contour of the microwell 195 becomes almost a perfect circle as shown by the solid line AM. The shape of the treatment egg 25 is similarly corrected and imaged in a shape close to its original shape. Not only shape distortion but also coloring due to chromatic aberration and the like are suppressed.

[0023] Next, the processes performed in the embryo culture device 10 will be described. FIG. 7 is an explanatory diagram showing the electrical configuration of the embryo culture device 10. As shown in the figure, a control unit 60 is provided in the embryo culture device 10. The control unit 60 includes a well-known CPU 61, ROM 62, RAM 63, a memory interface 64 for data exchange with the memory card 65, a general-purpose I / O interface 66 for signal exchange with external devices, a culture chamber interface 67 for signal exchange to control the environment in the culture chambers R11 to R24, a camera interface 68 for giving an imaging instruction and acquiring the captured image to / from the camera module 51, a video interface (abbreviated as video I / F) 69 for displaying an image on the display 70, and the like.

[0024] The CPU 61 uses a high-speed processor incorporating functions such as a DSP for rapidly processing fertilization determination based on an image described later, and a vector arithmetic function for performing determination processing using a neural network or the like. The ROM 62 stores a program for realizing the processes in the embryo culture device 10 including the time-lapse control process described later. The CPU 61 appropriately reads such a program from the ROM 62, expands it in the RAM 63, and executes it to realize necessary processes and controls. The memory card 65 records information on the culture environment, such as the temperature of each culture chamber R11 to R24, in addition to the images captured by the camera module 51. Instead of the memory card 65, a magnetic storage medium such as a hard disk or a semiconductor storage medium such as an SSD may be used. Alternatively, it may be stored in a so-called cloud connected via a network.

[0025] The display 70 has a touch panel provided on its surface, and by touching the display 70, buttons or the like displayed on the display 70 can be selected. In this embodiment, the display 70 is provided integrally with the embryo culture device 10, but it may be provided separately from the embryo culture device 10 and connected by wire or wirelessly. Alternatively, the embryo culture device 10 and a computer may be connected via a network or the like, and the display of the computer may be used as the display 70. Alternatively, a highly portable terminal such as a mobile phone or a tablet may be used as the display 70.

[0026] The general-purpose I / O interface 66 is connected to switches SW11 to SW24 provided in the embryo culture device 10, a drive device 71 for opening and closing the lid portion 17, a warning device 72 for generating a warning sound, a mixture adjusting device 73 for adjusting the air-fuel mixture, and the like. The mixture adjusting device 73 includes a pressure regulating valve for regulating the pressure of carbon dioxide gas, nitrogen gas, etc. supplied to the gas ports 18, 19, a pump for taking in the atmosphere through the filter 27, a pump for sending the air-fuel mixture into the culture chambers R11 to R24, and the like. Connected to the culture chamber interface 67 are panel heaters provided in the respective culture chambers R11 to R24, control valves V11 to V24 for controlling the supply amount of the air-fuel mixture, and the like. The control valves V11 to V24 are provided in the pipes leading from the air-fuel mixture supply pipe 84 for supplying the air-fuel mixture to the culture chambers R11 to R24.

[0027] A general-purpose I / O interface 66 is connected to a computer (hereinafter abbreviated as PC) 90 via a general-purpose connector 26. In addition to a display 91 and a keyboard (not shown), the PC 90 is provided with a rotary pointing device 95. The control device 95 includes a dial 96 with a smaller diameter on a substantially circular base portion 97, and by rotating the dial 96 left and right, it is possible to move a number of time-lapse images displayed on the display 91 forward and backward in time series for display. Such a device is also called a rotary selector. The dial 96 can not only be rotated left and right but also pushed in, and when selecting one of the currently displayed images, it realizes an "enter" operation similar to the operation by a mouse button. Of course, dedicated buttons may be provided on the base portion 97 to assign enter and selection operations. If time-lapse images are taken 24 hours every 10 minutes, there will be 6×24 = 144 images for each treated egg. When quickly referring to such a large number of images to confirm fertilization judgment, etc., such a rotary pointing device is useful.

[0028] (A-2) Embryo culture process: On the premise of the hardware configuration of the first embodiment described above, the embryo culture device 10 performs the following processes. In the default state where the embryo culture device 10 is powered on, the usage status of the culture units 11 to 24 of the embryo culture device 10 is displayed on the display 70. An example of this display is shown in column (A) of FIG. 8. As shown in the figure, nine rectangular regions 76 corresponding to the culture units 11 to 24 are displayed on the display 70, and among them, the used culture units are marked and displayed. Above the rectangular region 76, it is displayed that "When starting to use, please tap on the part of the culture unit to be used". In accordance with this instruction, when one of the regions displayed as not in use is tapped, the corresponding one of the culture units 11 to 24 becomes available. In the following description, the specified culture unit will be referred to with a suffix n, such as "culture unit n" and the corresponding culture chamber as "culture chamber Rn". Instead of tapping the touch panel on the display 70, various instructions may also be performed by operating the keyboard of the PC 90 connected to the general-purpose connector 26 or the control device 95.

[0029] When the switch SWn of the culture unit n selected by tapping the rectangular region 76 is pressed, the lid 17 opens, and the dish 191 can be accommodated in the culture chamber. Of course, the lid 17 may be opened only by tapping the rectangular region 76 of the display 70. When the dish 191 is placed in alignment with the holding frame 180 and the lid 17 is closed, the control unit 60 sets the temperature, gas concentration, etc. in the culture chamber Rn to a desired state, and then starts the time-lapse process and displays the information in the culture chamber Rn on the display 70. An example of this is shown in column (B) of FIG. 8. In this example, it is displayed that "Start culturing in culture chamber Rn", and below that, the current information of the culture chamber Rn is displayed. Specifically, information related to imaging such as the start time and elapsed time of imaging, the interval of imaging, the environment in the culture chamber, that is, temperature and humidity information, and further information such as gas concentration are displayed. The temperature and gas concentration are detected by sensors (not shown). In addition to such information, information identifying the patient of the treated egg, such as a medical record number, etc. may also be displayed.

[0030] Thus, when the cultivation is started, the 25 microwells 195 of the dish 191 in the cultivation chamber Rn are imaged together, divided and saved for each microwell 195, the fertilization possibility of each treated egg 25 in the microwell 195 is individually judged, and a time-lapse control process for displaying necessary images on the display 70 is performed. Among the time-lapse control processes, the fertilization determination process and the like will be briefly described later. For example, as shown in column (C) of FIG. 8, when the fertilization of each treated egg 25 is determined from the captured image, processes such as displaying this on the display 70 are performed.

[0031] The time-lapse control process will be described with reference to FIG. 9. FIG. 9 is a flowchart showing the time-lapse control process routine. This process is repeatedly executed at a predetermined interval when the use of the cultivation chamber Rn is started. When the time-lapse control process is started, first, it is judged whether a predetermined time has elapsed since the image of the treated egg was last captured (step S300). Here, the predetermined time corresponds to the imaging interval shown in column (B) of FIG. 10. Of course, it may be a shorter time or a longer time instead of the 10 minutes exemplified in column (B) of FIG. 10. If the predetermined time has elapsed since the last imaging (step S300: “YES”), the imaging process is performed (step S310). The imaging is performed by imaging a plurality of microwells 195 together, that is, the 25 microwells 195 existing in the dish 191 together. The captured image is saved in the memory card 65 together with the data of the imaging time. The imaging by the camera module 51 is performed from below the dish 191 as shown in FIG. 5. At this time, the camera module 51 may image a plurality of images for each microwell 195 while moving the focal position.

[0032] After performing the imaging process, the captured image is processed to be divided for each of the 195 microwells (step S330). Here, since there are 25 microwells 195 in the area captured by the camera module 51, the captured image is divided into 25 parts. Next, a process of correcting the divided image is performed (step S340). The correction is performed to remove distortion and dirt from the image. Since a plurality of microwells 195 are imaged at once, it is not possible to place all the microwells 195 directly below the optical axis of the camera module 51. In the present embodiment, as shown in FIG. 5, such distortion is reduced using the lens array LA which is the optical function unit 200, but still, slight distortion occurs in the image of each microwell 195. Also, there are cases where blurring or coloration due to aberration occurs.

[0033] Therefore, in the image correction (step S340), such distortion and dirt are corrected. When the distorted image is corrected, the contour of each microwell 195 becomes almost a perfect circle. The shape of the treatment eggs 25 is similarly corrected to its original shape. Such correction is performed not only to trim the distortion of the shape, but also to remove coloration due to chromatic aberration and ghosts due to lens dirt.

[0034] After the image correction, if correction can be performed, next, a process of saving the image to the memory card 65 is performed (step S350). At this time, the saving is performed for each of the divided images. Of course, each of the divided images may be tagged with, for example, a tag for identifying which microwell 195 it is and which culture chamber Rn it was captured in by the file name, etc., and associated. Also, at the time of saving, only the corrected image may be saved, or it may be saved together with the image before correction, or separately from the image before correction. The storage destination of the corrected image may be the same memory card 65 that stores the image before correction, or it may be saved to another storage device, such as a server on the cloud. The storage destination may be another storage medium, such as a hard disk or SSD different from the memory card 65.

[0035] In the series of processes described above, that is, from imaging by the camera module 51 to the processes of splitting, correcting, and then saving, the CPU 61 monitors whether any abnormalities have occurred (step Serr). The occurrence of an abnormality is determined at any time in the interrupt process.

[0036] When it is detected that some abnormality has occurred during the period from shooting to saving (steps S310 to S350) (step Serr), an abnormality display process (step S320) for displaying the details of the abnormality on the display 70 is performed. At this time, the display 70 functions as a presenting unit for presenting the abnormality. An example of the abnormality display is shown in column (D) of FIG. 8. In the illustrated example, as the details of the abnormality, □ Light lighting abnormality □ Camera malfunction □ Correction process abnormality □ Image splitting or saving abnormality etc. are exemplified. If these abnormalities occur, the corresponding locations are marked and displayed. If multiple overlapping abnormalities occur, multiple items are marked. In the example shown in column (D) of FIG. 8, "correction process abnormality" is marked.

[0037] Among the abnormalities, "light lighting abnormality" refers to the case where the illumination unit 170 does not light up, or when there is a bias in the illumination light and a part of the image is captured as a dark image. In such cases, the captured image becomes dark and the average brightness of the image does not fall within a predetermined range, so it can be discriminated. A camera malfunction is a case where the camera module 51 does not operate, such as when the light-receiving modules of the CCD or CMOS for shooting do not operate. Of course, when an optical focusing device or a shutter is provided, malfunctions of these devices are also included. Regarding the light-receiving module, for example, when the clock for reading is not input normally, it can be determined that there is a camera malfunction.

[0038] Correction processing abnormality refers to the case where correction for removing distortion or dirt in the captured image cannot be performed normally. For example, for distortion, the distance from the optical axis, etc. is input in advance as a parameter, and shape correction, etc. is performed using this parameter to remove it. However, if as a result of the correction, the shape of the microwell 195 is significantly different from a perfect circle, it is determined that the correction processing could not be performed normally. Also, the process of removing dirt is usually performed by removing one or two isolated dots on the screen. However, if the number of isolated dots after the correction process is equal to or more than a predetermined number, it is determined that the correction process could not be performed normally.

[0039] Among image division or storage abnormalities, division abnormality refers to the case where when dividing an image of a plurality of microwells 195, the division process does not end normally, or due to reasons such as the position of the dish 191 being displaced, the imaging position by the camera module 51, etc. is displaced, and each microwell 195 does not fit into each of the divided images after division. Storage abnormality of the divided image refers to the case where when saving the file of the divided image to a memory card 65, etc., it cannot be saved normally due to insufficient capacity or failure of the memory card 65, etc.

[0040] After finishing the above processing (steps S330 to S350), if there is no particular abnormality, it is determined whether it is the timing when fertilization can be judged (step S360). Judging fertilization means judging whether fertilization has occurred in the treated egg 25 on which the fertilization treatment has been performed. Fertilization usually occurs within about 7 hours at the earliest, on average about 17 hours, and even in the latest case, within 24 hours after the fertilization treatment. The treated egg 25 on which the fertilization treatment has been performed is immediately accommodated in the microwell 195 of the dish 191 after the treatment and placed in the culture units 11 to 24. Therefore, when about 6 hours have passed since the imaging by the camera module 51 started, the judgment in step S360 is "YES", that is, fertilization can be judged. Of course, this time can be made shorter, and fertilization can be judged immediately after the dish 191 is accommodated in the culture rooms R11 to R24. In that case, the judgment in step S360 is unnecessary.

[0041] When it is determined that fertilization judgment is possible, next, a fertilization judgment process is performed (step S370). This process can be determined by reading out the images of the treated eggs 25 stored in the memory card 65 after being divided for each microwell 195 and checking whether two pronuclei are clearly shown in the images. Of course, this judgment may be made based on reading out a large number of images previously stored in the memory card 65 and based on the changes over time, particularly the change in the number of sites recognized as pronuclei. Further, a large number of images of fertilized treated eggs and images of unfertilized treated eggs may be prepared and machine-learned in advance, and the images read out from the memory card 65 may be input into this machine-learned determination machine to determine the possibility of fertilization. Such a process can be realized in real time by utilizing the high-speed processing function of the CPU 61 shown in FIG. 7. Incidentally, the captured images may be displayed on the display 70 to allow the embryo culturist to determine the possibility of fertilization.

[0042] After executing the fertilization judgment process (step S370), it is determined whether the treated egg 25 is a fertilized egg for which fertilization can be confirmed (step S380). If it can be determined that it is a fertilized egg, a fertilized egg number display (step S390) is performed. An example of this display has already been described with reference to column (C) of FIG. 8. In this example, a message 77a indicating that fertilization has been confirmed, an image of the treated egg 25 for which fertilization has been confirmed, a number display 77b indicating which microwell 195 the fertilized egg is in, and an "END" button 78 for giving an instruction to end the screen display are displayed on the display 70. Thereafter, it proceeds to "NEXT" to end this processing routine.

[0043] If photography is started and it is assumed that there is a fertilized egg and there are no particular abnormalities until it is displayed, in the example of FIG. 8, the display from column (A) through column (B) to column (C) is performed. If any abnormality occurs from photography to storage, the display in column (D) is performed. Note that the detection of an abnormality does not need to be performed for all the items described above, and it may be performed for at least any one of them. Of course, the detection and notification of an abnormality other than the items described above may be performed. In such a case, the notification of an abnormality may be a display on the display 70, or may be an audio notification using an audio output device (not shown). Also, it may be a mode of notifying the person in charge by mail or message to a mobile phone or the like.

[0044] In the first embodiment described above, since the lens array LA is formed as the optical function unit 200 at the bottom of the microwell 195 of the dish 191, a part of the lens in the lens module 52 of the camera module 51 on the embryo culture device 10 side can be omitted, and the lens module 52 of the embryo culture device 10 can be configured compactly. Also, the positional relationship between each lens of the lens array LA and the microwell 195 can be fixed, and the alignment of each lens of the lens array LA at the time of imaging can be made reliable. The dish 191 provided with the microwell 195 is usually placed by the user's hand in the holding frame 180 of the culture chamber R11, so its placement position may not be exactly the same. Even in such a case, since the optical axes of each lens of the lens array LA and the microwell 195 are aligned from the beginning, there is no need to perform the adjustment of aligning the optical axes one by one. Also, when the dish 191 is placed by an arm such as a robot, it is not necessary to make the positioning accuracy very strict, and it is easy to realize the placement of the dish 191 by the robot.

[0045] Furthermore, in the first embodiment, since the camera module 51 is fixedly provided for each of the culture units 11 to 24, it is not necessary to move the dish 191 when imaging the treated eggs 25. Therefore, the imaging interval in time-lapse can be shortened, or continuous video shooting can also be performed. In addition, since the microwells in which a maximum of 25 treated eggs 25 are arranged in a narrow range are adopted, the imaging range by the camera module 51 can be narrowed, and the resolution per treated egg of the captured image can be increased. Moreover, by arranging the lens array LA directly below the microwell 195, the viewing angle of each microwell 195 as seen from the camera module 51 can be reduced, and the distortion of the image of the treated egg captured can be reduced. For this reason, while imaging a maximum of 25 microwells 195 and the treated eggs 25 accommodated therein at once, it is possible to improve the accuracy of various determinations based on the images separately stored for each treated egg 25, such as fertilization determination. Further, in this embodiment, since the images of each treated egg are corrected before determination, distortion, dirt, etc. can be removed by the correction, and the accuracy of the determination can be further improved.

[0046] B. Second Embodiment: Next, the second embodiment will be described. The embryo culture apparatus and dish of the second embodiment have the same configuration as the first embodiment. However, as shown in FIG. 10, the camera module 51 and the lens module 52 are provided in the culture chamber Rn at positions for imaging the dish 191 from above, and the dish 191 is different in that a diffuser plate 201 is provided as an optical functional part. To image the microwell 195 of the dish 191 from above, the camera module 51 or the like may be provided on the lid part 17. Further, the illumination unit 170 may be provided at the position of the camera module 51 in the first embodiment, that is, below the bottom of the dish 191.

[0047] In the second embodiment, a diffusion plate 201 is provided directly below the microwells 195 as an optical functional part at the bottom 196 of the dish 191. A lighting unit 170 is provided below the diffusion plate 201. The lighting unit 170 is provided with high-brightness LEDs 175. The light emitted from the high-brightness LEDs 175 in the direction of the microwells 195 is diffused by the diffusion plate 201 to become uniform light, which illuminates the microwells 195. By providing the diffusion plate 201 directly below the microwells 195, a light source that is less likely to create shadows, like a shadowless lamp, is prepared while using a single high-brightness LED 175, and the treatment eggs 25 accommodated in each microwell 195 can be evenly illuminated.

[0048] The treated egg 25 illuminated from below by such a light source is imaged by the camera module 51 provided on the side opposite to the illumination unit 170. At this time, the position of the lens provided in the lens module 52 is moved to move the focal position in the vertical direction. This will be described with reference to FIG. 10. By moving at least one of the lenses provided in the lens module 52 up and down along the optical axis, the focal position of the camera module 51 can change to positions GL+1, which is farther from the camera module 51 than the central position GL0, and even farther position GL+2, and positions GL-1, which is closer to the camera module 51, and even closer position GL-2. Since the size of the treated egg 25 is about 100 to 200 μm if it is a human egg, the difference between each position is set to approximately 20 to 25 μm. In this case, the lens configuration of the lens module 52 has a shallow depth of focus, and the imaging range at each focal position is focused at 25 to 30 μm. When imaging the treated egg 25 using the camera module 51, the focal position is changed from GL-2 to GL+2, and five images are captured. Of course, more images (for example, about 11 images) may be captured by changing the focal position. If a plurality of images with different focal positions are captured, when making a judgment on fertilization in the treated egg 25, the pronucleus, which serves as a clue, is likely to be clearly shown in one of the plurality of images, regardless of where it is located in the treated egg 25, which is substantially spherical. Also, even if the two pronuclei overlap when viewed from the camera module 51 side, they can be observed separately.

[0049] In the second embodiment having such a configuration, in addition to achieving the same operational effects as the embryo culture device of the first embodiment, as an optical function unit, a diffusion plate 201 is provided at the bottom 196 directly below the microwell 195, so that the light diffused by the diffusion plate 201 can be applied to the microwell 195. Therefore, an image of the treated egg 25 accommodated in the microwell 195 can be captured with uniform illumination light.

[0050] C. Third Embodiment: Next, the third embodiment will be described. The embryo culture device of the third embodiment and the dish installed therein have the same configuration as that of the first embodiment, but differ in that a light guide path is provided in the dish as an optical functional unit. As shown in FIG. 11, the dish 191 of the third embodiment has a shape in which a part of the circular outer shape is linearly cut. When the dish 191 is installed in accordance with the linear portion 183 provided in the holding frame 180, the light receiving portion 199 of the dish 191 is in contact with the light emitting portion 81 provided in the linear portion 183.

[0051] As shown in FIG. 12, the light emitting portion 81 is connected to the general-purpose I / O interface 66 in the control unit 60, and the five light emitting diodes 231 to 235 provided in the light emitting portion 81 are sequentially lit in accordance with the instruction of the CPU 61. At the light emitting surfaces of the light emitting diodes 231 to 235 of the light emitting portion 81, the ends of the light guide paths 221 to 225 provided in the light receiving portion 199 of the dish 191 are opposed. Therefore, when each of the light emitting diodes 231 to 235 is lit, the light is guided to the light projecting rings 211 to 215 on the outer periphery of the microwell 195 through the light guide paths 221 to 225.

[0052] In FIG. 12, since the microwell 195 is viewed from above, the light projecting rings 211 to 215 are drawn overlapping, but as shown in FIG. 13, the light projecting rings 211 to 215 are arranged to overlap in the height direction. Further, a similar structure is provided on the outer periphery of the 5×5 microwells 195. Therefore, when any one of the light emitting diodes 231 to 235 emits light, light is emitted into the microwell 195 from any one of the corresponding light projecting rings 211 to 215 provided on the outer periphery of all the microwells 195.

[0053] As shown in FIG. 13, each of the light projecting rings 211 to 215 is composed of a core portion 252 having a predetermined refractive index and a clad portion 251 covering the core portion and having a refractive index higher than that of the core portion 252. The configuration of such a core portion and clad portion is the same for the light guide paths 221 to 225. Therefore, the light of each of the light emitting diodes 231 to 235 incident from the ends of the light guide paths 221 to 225 is totally reflected at the boundary between the core portion 252 and the clad portion 251, and is efficiently guided to the end faces on the micro well 195 side of each of the light projecting rings 211 to 215, and is emitted from here to illuminate the treatment egg 25.

[0054] Therefore, if the CPU 61 shifts the lighting timings of the light emitting diodes 231 to 235 provided in the light emitting unit 81 and drives the camera module 51 in accordance with the lighting timings of the light emitting diodes 231 to 235 to perform imaging, images up to a plurality of positions GL-2 to GL+2 of the treatment egg 25 can be captured without adjusting the focus position by the lens module 52. In this case, if the depth of focus realized by the lens of the lens module 52 is increased so that it is, so to speak, a single focus lens configuration, any image can be captured without being blurred.

[0055] According to the third embodiment, as the optical function unit 200, the light guide paths 221 to 225 for guiding the light of the light emitting diodes 231 to 235 and the light projecting rings 211 to 215 for emitting the guided light at different heights of the micro well 195 are provided. On the embryo culture device 10 side, images at different positions in the height direction within the micro well 195 can be captured only by controlling the lighting timings of the five light emitting diodes 231 to 235. Therefore, it is not necessary to mount a mechanism for moving the lens in the lens module 52 to adjust the focusing position. For this reason, it is not necessary to consider the moving time of the lens, etc., and the imaging interval in the time-lapse control can be shortened. In addition, since there is no movable part, failures etc. are less likely to occur, and since lubricating oil etc. for movement is not necessary, the mixer introduced into the culture chambers R11 to R24 is not contaminated with evaporated lubricating oil etc. The dish 191 of the third embodiment can also exhibit the same functions and effects as those of the first and second embodiments, such as simplifying the configuration of the embryo culture device 10.

[0056] D. Fourth Embodiment: The configuration of the fourth embodiment is illustrated in FIG. 14. In the figure, it is depicted that two optical functional components are mounted on the dish 191, but only one of them may be sufficient. Also, it is possible to use them in combination with the optical functional components of the first to third embodiments. In this example, a marking 310 as an optical functional component is provided on the bottom surface 196 of the dish 191, and this is imaged by a camera 351 provided on the lid portion 17. The marking 310 optically has a function of transmitting / non-transmitting light, and is formed by engraving with a laser on the bottom surface 196 of the dish 191. For example, assuming 4×4 dots, at least 3 of the 4 corners among them are engraved in the form of laser light, that is, as black dots, and at least 12 other dots are set as engrave = on (1), not engrave = off (0), so that 2 12 types of marks can be formed. Therefore, by attaching any one of the marks to the dish 191, the dish 191 can be distinguished. Moreover, in this case, since it is a mark engraved on the dish 191 rather than pasting a mark with a tape or a label, there is no risk of mis-pasting a label or the like. The dish 191 may be provided with the marking 310 from the beginning, but a low-output laser output may be provided in the embryo culture apparatus 10, and when it is used for the first time, the patient's ID etc. may be written on the spot.

[0057] Similarly, a marking 320 as an optical functional component may be provided on the outer peripheral wall 194 of the dish 191. In this case, a camera 352 may be provided on the side wall of the culture chamber facing the marking 320 to image the marking 320. The dish 191 is provided with a recess 197 on the outer peripheral wall 194. As shown in FIGS. 2 and 3, in relation to the protrusion 182 provided on the holding frame 180, its position is uniquely determined, so it is easy to provide the camera 351 or the camera 352 at a position where the marking 310 or the marking 320 can be imaged. Note that the marking can also be easily formed by printing with an inkjet printer in addition to engraving with a laser beam.

[0058] Also, by making the outer peripheral wall 194 opaque and making holes in it with a laser beam or a micro drill, or melting the surface to make it transparent, it is easy to perform marking. To make the outer peripheral wall 194 opaque, it may be painted, an opaque film may be formed, or the surface may be roughened. The marking may be formed in a form distinguishable by visible light, or may be formed in a form distinguishable by light other than visible light, such as infrared light or ultraviolet light. By laminating a film with a film thickness of 1 / 2 of the wavelength for light of a specific wavelength, the transmission of light of that wavelength can be reduced. If such a film is laminated on the surface of the dish 191 and melted and evaporated with a laser beam, it is easy to form a mark that can be read by light of such a specific wavelength. If the markings 310, 320 are made to be readable by light of a specific wavelength, it becomes difficult to modify the markings, and the security of the embryo culture device 10 can be improved.

[0059] The markings 310, 320 are not limited to the dot patterns described above, and may be two-dimensional barcodes, one-dimensional barcodes, etc., as long as they are formed by an optical function. Also, simply an image, for example, a patient's autograph, etc. may be used. Such an image may be formed by a hologram or the like. Whether the image matches the registered image may be determined by dynamic pattern matching, or the images in a dictionary or the like may be learned by machine learning or the like and then determined.

[0060] E. Regarding variations of the embodiment: The microwells 195 are not limited to an array of 5×5, and may have an even smaller number of arrays, or may be arranged in other array forms, such as in a circular shape. As long as the required resolution is obtained, any number of treated eggs can be imaged at once. The content and method of correction and image segmentation may be changed according to the arrangement of the microwells 195. Also, a plurality of camera units fewer than the number of dishes 191 may be provided and moved within the case body 20 so that the camera units share the photographing of a plurality of trays.

[0061] In the embodiment described above, it was determined whether the treated egg 25 was fertilized. However, it is also possible not to make such a determination, or after making the determination, continue to culture the treated egg 25 and image its state with the camera module 51. Alternatively, it may be configured as a device that automatically performs all operations from the step of accommodating the treated egg 25 in the microwells 195 of the dish 191 to the step of sending out the fertilized egg for the next treatment.

[0062] The camera module 51 was prepared for each dish 191. However, two or more camera modules 51 may be prepared for one dish 191, and a plurality of microwells 195 in one dish 191 may be imaged with separate camera modules 51. The plurality of camera modules 51 may overlap in imaging the plurality of microwells 195. In this way, even if one camera module fails and imaging cannot be performed, images of all the microwells 195 can be obtained by using the images of other camera modules.

[0063] In each of the above embodiments, all the wells were described as microwells. However, those in which the treated egg 25 is accommodated in normal wells larger than this may also be used. Also, the shape of the well only needs to be a depression capable of accommodating the treated egg 25, and is not limited to a hemispherical shape, and may be a depression having an elliptical cross-sectional shape, an oval-shaped depression, or the like.

[0064] F. Other aspects: (1) One aspect of the present disclosure is an aspect as a dish used in an embryo culture device for culturing embryos. This dish is formed of a transparent resin or glass, and a plurality of wells, which are depressions having a diameter capable of accommodating treated eggs, are formed at the bottom of the dish, and an optical functional portion having a predetermined optical function is formed on at least one of the bottom side and the side portion side of the well. By doing so, since the optical functional portion is formed on a part of the dish, a part of the optical device on the embryo culture device side can be omitted, and the embryo culture device can be configured compactly. Further, since the optical functional portion is provided on the side of the dish, when the optical functional portion is soiled or the like, it is only necessary to replace the dish, and the need to replace or repair the optical device on the embryo culture device side is suppressed.

[0065] (2) In such a configuration, the optical functional portion may be at least one of [1] a lens provided directly below the well, [2] a light guide path provided at a position surrounding the outer periphery of the well, and [3] an optical filter provided directly below the well. By doing so, in the case of [1], a part of the lens on the embryo culture device side can be omitted, and the positional relationship between the necessary lens and the well can be easily determined. In the case of [2], light guiding of illumination light to the well can be easily realized. Further, by providing the light guide path in the dish, the embryo culture device can be made compact. On the other hand, in the case of [3], the optical filter can be provided close to the well, the optical filter can be made small, and its function can be sufficiently exhibited. The optical filter includes, for example, those that transmit or block light of a specific wavelength, and a diffusion plate that diffuses light. As the light of a specific wavelength, when the optical filter is provided on the side that irradiates the light in the well, light of a wavelength that may affect the treated eggs accommodated in the well, such as ultraviolet light, is assumed. When the optical filter is provided on the side for imaging, it is conceivable to use an optical filter that selectively transmits light of a wavelength meaningful for knowing the state of the treated eggs, such as light near infrared that is normally emitted by a living body.

[0066] (3) In such a configuration, the optical functional unit is an optical path provided at a position surrounding the outer periphery of the well. The optical paths are stacked in plurality in the thickness direction of the well. The input-side end of the optical path may be provided at a position facing an LED for illumination provided for each of the optical paths. By doing so, illumination light can be selectively projected to different positions in the thickness direction of the well, and for example, imaging at different positions in the thickness direction of the treated egg becomes easier.

[0067] (4) A second aspect of the present disclosure is an aspect as an embryo culture device. One of such embryo culture devices includes a dish having the above-described optical functional unit, a culture unit that houses the dish and provides a culture environment for the embryo, and an imaging unit that is fixed in position in the culture unit and images the well from above or below, and the imaging unit performs imaging using the optical functional unit when imaging the well. According to this embryo culture device, since the dish has an optical functional unit having various functions, it is not necessary to provide a member corresponding to the optical function on the embryo culture device side, and the configuration of the embryo culture device can be simplified.

[0068] (5) In such a configuration, it may be a dish having an optical path on the outer periphery of the above-described well, a culture unit that houses the dish and provides a culture environment for the embryo, an LED for illumination provided at each input-side end of the plurality of optical paths, an imaging unit that is fixed in position in the culture unit and images the well from above or below, and an imaging control unit that controls lighting of the plurality of LEDs to perform imaging in two or more lighting states with different lighting states when imaging the well by the imaging unit. By doing so, the well can be easily imaged as two or more lighting states with different lighting states, and the range of use of the captured image can be expanded. This is because images in such different lighting states are highly useful when determining the state of the treated egg.

[0069] (6) In such a configuration, the different illumination states may be illumination states that enable imaging of an image centered on different positions in the height direction of the treatment eggs accommodated in the well via the plurality of light guide paths. By doing so, a camera that captures an image two-dimensionally from one direction can capture an image of the treatment eggs in the well while focusing on different positions in the height direction of the treatment eggs, and for example, the accuracy of fertilization determination of the treatment eggs can be improved.

[0070] (7) A third aspect of the present disclosure is an aspect as a method for imaging an embryo. This method for imaging an embryo is a dish formed of a transparent resin or glass and having a plurality of wells, which are depressions with a diameter capable of accommodating treatment eggs, formed at the bottom, and an optical functional part having an optical function is formed on at least one of the bottom side and the side part side of the well. The dish is held in a culture part that realizes a culture environment for the embryo, and imaging is performed using the optical functional part of the dish by an imaging part that is fixed in position in the culture part and images the well from above or below. By doing so, the treatment eggs in the well can be imaged by the optical functional part on the dish side and the imaging part. In this case, since the optical functional part is provided on the dish side, the distance between the optical functional part and the treatment eggs can be reduced, and the optical function of the optical functional part can be utilized.

[0071] (8) In each of the above embodiments, a part of the configuration realized by hardware may be replaced with software. At least a part of the configuration realized by software can also be realized by a discrete circuit configuration. Further, when a part or all of the functions of the present disclosure are realized by software, the software (computer program) can be provided in a form stored in a computer-readable recording medium. The "computer-readable recording medium" includes not only portable recording media such as flexible disks and CD-ROMs, but also various internal storage devices in a computer such as various RAMs and ROMs, and external storage devices fixed to a computer such as hard disks. That is, the "computer-readable recording medium" has a broad meaning including any recording medium capable of fixedly storing data packets not temporarily.

[0072] The present disclosure is not limited to the above-described embodiments, and can be realized in various configurations without departing from the gist thereof. For example, the technical features in the embodiments corresponding to the technical features in each form described in the summary of the invention can be appropriately replaced or combined in order to solve some or all of the above-described problems or to achieve some or all of the above-described effects. Further, if the technical feature is not described as essential in this specification, it can be appropriately deleted.

Explanation of Reference Numerals

[0073] 10…Embryo culture device, 11…Culture unit, 17…Cover part, 18…Gas port, 20…Case body, 21…Culture unit, 25…Treated egg, 26…General-purpose connector, 27…Filter, 28…Filter port, 43…Supply port, 44…Exhaust port, 51…Camera module, 52…Lens module, 53…Image sensor, 60…Control unit, 61…CPU, 62…ROM, 63…RAM, 64…Memory interface, 65…Memory card, 66…General-purpose I / O interface, 67…Culture chamber interface, 68…Camera interface, 70…Display, 71…Drive device, 72…Warning device, 73…Mixture adjustment device, 76…Rectangular area, 77a…Message, 77b…Number display, 78…Button, 81…Light-emitting part, 84…Mixture supply pipe, 90…Computer (PC), 91…Display, 95…Control device, 96…Dial, 97…Base part, 170…Illumination unit, 180…Holding frame, 182…Projection, 183…Straight part, 191…Dish, 192…Partition wall, 194…Outer peripheral wall, 195…Microwell, 196…Bottom part, 197…Recess, 198…Opening, 199…Light-receiving part, 200…Optical function part, 201…Diffusion plate, 211~215…Light-projecting ring, 221~225…Light guide path, 231~235…Light-emitting diode, 251…Clad part, 252…Core part, 310,320…Marking, 351,352…Camera

Claims

1. A dish used in an embryo culture apparatus for culturing embryos, wherein the dish is formed of a transparent resin or glass, a plurality of wells, which are depressions having a diameter capable of accommodating treated eggs, are formed at the bottom of the dish, an optical functional part having a predetermined optical function is formed on at least one of the bottom side and the side part of the well, the optical functional part is an optical path provided at a position surrounding the outer periphery of the well, a plurality of the optical paths are laminated in the thickness direction of the well, an input side end of the optical path is provided at a position facing an LED for illumination provided for each of the optical paths, a dish.

2. The dish according to Claim 1, wherein the optical functional part further includes, a lens provided directly below the well, and an optical filter provided directly below the well, a dish including at least one of them.

3. The dish according to Claim 1 or Claim 2, and a culture unit that houses the dish and provides a culture environment for the embryos, an imaging unit that is fixed in position in the culture unit and images the well from above or below, and uses the optical functional part for imaging when imaging the well, an embryo culture apparatus comprising the same.

4. The dish according to Claim 1, and a culture unit that houses the dish and provides a culture environment for the embryos, LEDs for illumination provided at input side ends of the plurality of optical paths, an imaging unit that is fixed in position in the culture unit and images the well from above or below, an imaging control unit that controls lighting of the plurality of LEDs when imaging the well by the imaging unit, and performs imaging in two or more lighting states with different lighting states, an embryo culture apparatus comprising the same.

5. The embryo culture apparatus according to Claim 4, wherein the different lighting states are lighting states in which images can be taken centering on different positions in the height direction of the treated eggs accommodated in the well through the plurality of optical paths.

6. A dish formed of a transparent resin or glass, having a plurality of wells, which are depressions having a diameter capable of accommodating treated eggs, formed at the bottom, and an optical functional part having an optical function is formed on at least one of the bottom side and the side part of the well, and the dish is held in a culture unit for realizing a culture environment for embryos, As the optical function unit, a plurality of light guide paths are laminated in the thickness direction of the well at a position surrounding the outer periphery of the well, and the lighting of the LED provided for each light guide path is controlled at a position facing the input side end of the light guide path. In the culturing unit, imaging is performed using the illumination of the LED guided through the light guide path of the dish by an imaging unit that is fixed in position and images the well from above or below. Imaging method for embryos.

Images