JP2018093795A - Embryo culturing apparatus and imaging device therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To store a large number of trays holding treated eggs in a limited space and implement time lapse processing.SOLUTION: An embryo culturing apparatus which stores a plurality of trays holding treated fertilized eggs and maintains same in a culture environment is provided with a culture unit that maintains the culture environment, and a plurality of imaging units provided to respond to each one of the trays held in the culture unit that captures images of where the tray is present continuously or intermittently. The images captured by each of the imaging units are stored in a storage unit for each tray, and the stored images for each tray are displayed on a display unit.SELECTED DRAWING: Figure 2

Description

  The present invention relates to a technique for culturing an embryo.

  Social demands for infertility treatment are increasing, and various devices for culturing treated eggs that have been fertilized under a microscope have been proposed or sold. In recent years, a device is also widely used in which a camera is provided in such a culture apparatus to image a treated egg during culture and display the captured image on an external display as necessary (see Patent Document 1 below). . In such an embryo culture apparatus, the treated egg is placed in a tray (also referred to as a dish) that stably holds the egg, and the tray is transported into the field of view of the camera at a predetermined interval and imaged. By observing this image, the embryo cultivator can determine whether or not fertilization is possible, so that the treated egg does not need to be taken out of the culture apparatus only for observation.

JP 2012-39929 A

  On the other hand, there is a demand for culturing a large number of treated eggs in one embryo culturing apparatus in accordance with the rise of infertility treatment, and a plurality of trays are housed in the embryo culturing apparatus. Then, the following problems occur when imaging the treated egg subjected to fertilization treatment. Such a problem arises not only when fertilized eggs that have undergone microinsemination but also when cultured eggs that have been subjected to in vitro fertilization by some method.

[1] The time interval for imaging one treated egg becomes longer. For example, if it takes 2 minutes to transport and image a tray on which one treated egg is held, in an apparatus containing 10 trays in one embryo culture apparatus, 20 per tray. Only one image can be obtained per minute. As the number of trays stored is increased, the transport path to the imaging device becomes longer and takes more time, so the interval from imaging to the next imaging increases geometrically. For this reason, the number of trays that can be accommodated in one embryo culture apparatus is limited.

[2] The structure for transporting the tray becomes complicated and easily breaks down. If the conveyance mechanism malfunctions and the conveyance is stopped, the tray is placed outside the original culture space, usually a culture space maintained in a constant temperature / humidity environment, and is susceptible to adverse effects. Of course, if a constant temperature / humidity environment including the conveyance path and the imaging location is maintained, such adverse effects are minimized, but there is a problem that the apparatus configuration is easily increased in size.

[3] Since the treatment eggs in a plurality of trays are imaged by one imaging device, management of the captured images is complicated. Since complicated management is easy to make mistakes, tags and barcodes corresponding to patient IDs have been conventionally provided on trays, and these tags and barcodes are imprinted on the captured image along with the image of the treated egg, resulting in a mix-up. The device has been devised so as not to occur. Even in this case, if there is an error in reading a tag or barcode, an administrative error is still induced.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following aspects or application examples.

(1) As one embodiment of the present invention, an embryo culturing apparatus is provided that accommodates a plurality of trays that accommodate treated eggs that have been fertilized and holds them in a culture environment. The embryo culture apparatus includes a culture unit that holds the plurality of trays in the culture environment and a plurality of imaging units, and is provided corresponding to each of the trays held in the culture unit. An imaging unit that continuously or intermittently images the existing portion of the tray, a storage unit that stores an image captured by each imaging unit for each tray, and the stored each tray A display unit for displaying an image. According to this embryo culture apparatus, an imaging unit is provided corresponding to each of the plurality of trays held in the culture environment, and the captured images are stored for each tray. There is no need to move the tray or the imaging device to image the treated egg. As a result, a large number of trays can be arranged in one embryo culture apparatus, and a large number of embryo cultures can be performed at a time. As a result, it can contribute to infertility treatment.

(2) In such an embryo culture apparatus, the tray may include a plurality of recesses for holding the treated eggs, and the imaging unit may image at least a part of the plurality of recesses at a time. If it carries out like this, a some treatment egg can be accommodated in one tray, and the one part can be imaged at once. When the imaging unit can capture only a part of the plurality of recesses at a time, a plurality of imaging units may be prepared for one tray in order to capture all the recesses.

(3) In such an embryo culture apparatus, the culture unit may be composed of a plurality of culture chambers partitioned for each tray. If the culture section is partitioned for each tray, the temperature, humidity, gas concentration, etc. can be stabilized in a range partitioned for each tray, and the control of the culture section can be simplified. . Of course, the culture chamber is not particularly partitioned, and a plurality of trays may be held together.

(4) In such an embryo culture apparatus, the imaging unit may be provided on at least one of the upper surface, the side surface, and the bottom surface of the culture chamber so as to face the site where the tray is present. If arranged on the upper surface, the imaging unit can function as a lid of the culture chamber. In addition, the maintainability of the imaging unit is excellent. On the other hand, if it is arranged at the bottom, wiring to the imaging unit can be easily performed. In addition, it is easy to image the treated egg in the tray. On the other hand, the arrangement on the side surface can narrow the interval between the trays, and a large number of trays can be arranged in the embryo culture apparatus.

(5) In this embryo culture apparatus, the imaging unit includes an imaging depth-of-focus changing unit that captures a plurality of images having different in-focus positions in the imaging, and the storage unit has different in-focus positions. A plurality of images may be stored as a group of images. By doing this, it is possible to image the state inside the treated egg and accurately determine the status of the treated egg, for example, whether or not it has been fertilized.

(6) In the embryo culture apparatus, the display unit may include an individual display unit provided for each of the plurality of trays corresponding to the imaging unit. In this way, since the image captured by the imaging unit can be displayed as it is, there is no need to switch the display image. Of course, a smaller number of display units than the number of imaging units may be provided for a plurality of imaging units, and images captured by the plurality of imaging units may be switched and displayed.

(7) In such an embryo culture apparatus, the individual display unit may be provided on an upper surface of an openable / closable lid provided in the culture unit. By doing so, the relationship between the display and the treated egg accommodated in the culture part becomes clearer.

(8) The embryo culture apparatus described above may include a light guide that guides light from a light source and illuminates at least a portion where the treated egg exists in each of the plurality of trays. If it carries out like this, the object of the imaging by an imaging part can be illuminated and imaging of a treatment egg will become easy.

(9) Furthermore, in such an embryo culture device, a fertilization determining unit that determines whether at least one of the treated eggs in the tray has been fertilized based on an image captured by the imaging unit; Provided corresponding to the plurality of trays that can be held, and when the fertilization determining unit determines that at least one of the treated eggs has been fertilized, the tray for storing the treated eggs determined to have been fertilized is identified. It is good also as a person provided with the alerting | reporting part which alert | reports.

(10) In the embryo culture apparatus, the imaging unit further includes an instruction unit that instructs the imaging units to start imaging individually. Each of the imaging units receives an instruction from the instruction unit. In response, the imaging may be performed at independent timings. By doing so, the timing of imaging by the plurality of imaging units in the embryo culture apparatus can be made not to overlap, so that an embryo culture apparatus containing a large number of trays can be handled by a small number of embryo cultivators. Note that the imaging timing may be synchronized for each predetermined number of imaging units.

(11) As a second aspect of the present invention, there is provided an imaging device attached to a culture unit of an embryo culture device that accommodates a plurality of trays that contain fertilized treatment eggs and holds them in a culture environment. The imaging apparatus corresponds to each of the plurality of trays held in the culture unit, and includes a plurality of imaging units that continuously or intermittently image the existing portions of the trays, and the imaging units. A storage unit that stores the image captured for each tray, and a display unit that displays the stored image for each tray. This imaging apparatus for an embryo culture apparatus can be attached to an existing embryo culture apparatus and replaced with an apparatus for imaging and storing treated eggs. For this reason, the existing embryo culture apparatus can be used effectively.

  The present invention can be implemented not only as such an embryo culture apparatus and an imaging apparatus for an embryo culture apparatus but also in other modes. For example, an embryo culture method, a method of attaching an imaging device to an embryo culture device, a method of manufacturing an embryo culture device, a method of observing a treated egg contained in the embryo culture device, or a fertilization determining device for a treated egg and its method be able to.

The top view of the embryo culture apparatus as 1st Embodiment. The perspective view which shows a part of embryo culture apparatus. The top view which illustrates one of the culture chambers. Explanatory drawing which shows the electrical structure of the embryo culture apparatus containing a time lapse part. The top view which shows an example of the tray which accommodates a treatment egg. Explanatory drawing explaining the relationship between a time lapse part and the well of a tray. Explanatory drawing which shows the internal structure of a time lapse part. The flowchart which shows a culture | cultivation unit control routine. Explanatory drawing which shows the example of a display of a display module. The flowchart which shows a time lapse control processing routine. Explanatory drawing which shows an example of imaging of the treatment egg in a well. The schematic diagram which shows the principle which adjusts the position of a focus in the case of the imaging by a camera unit. Explanatory drawing which shows the electrical structure of the embryo culture apparatus containing the time lapse part in 2nd Embodiment. The flowchart which shows the main control routine in 2nd Example. The schematic diagram which shows the structural example at the time of providing a camera unit in the side surface of a culture chamber. Explanatory drawing which shows the relationship between the well at the time of seeing a tray from the side surface, and the treatment egg accommodated in the well.

A. First embodiment:
A first embodiment of the present invention will be described. FIG. 1 is a plan view of an embryo culture apparatus (also called an incubator) 10, and FIG. 2 is a perspective view showing a part of the embryo culture apparatus 10. The embryo culture apparatus 10 is for culturing an ovum that has been treated for in vitro insemination (hereinafter referred to as a treated egg) in a predetermined culture environment, that is, constant temperature and humidity for a certain period. Since the treated egg is not necessarily a fertilized egg, it is cultured in the embryo culture apparatus 10 for a predetermined period. The treatment of in vitro fertilization may be microinsemination performed under a microscope, or 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.

  As shown in FIG. 1, the embryo culture apparatus 10 includes 12 culture units 11 to 22. The configuration and operation of each culture unit 11-22 will be described later. The culture units 11 to 22 are provided on the flat case body 9 in two rows in the front and rear and six in the width direction. Each of the culture units 11 to 22 is provided with switches SW11 to SW22 correspondingly. By operating these switches SW11-22, the time lapse part 50 of the corresponding culture units 11-22 is opened and closed by a drive mechanism (not shown). Of course, the culture units 11 to 22 may be manually opened and closed.

  FIG. 2 shows a state in which the time lapse unit 50 is opened in the culture unit 12. The time lapse unit 50 includes a camera module 51 and a display module 52. The camera module 51 and the display module 52 have the same size and are integrally formed. The camera module 51 is provided with four camera units CA1 to CA4. These structures and functions will be described later. Culture units R11 to R22 (see FIG. 3) are prepared for each of the culture units 11 to 22, and the time lapse part 50 of each of the culture chambers R11 to R22 corresponds to the corresponding culture chambers R11 to R22. Also works as a lid. A silicone rubber seal portion is provided on the outer periphery of the time lapse portion 50 on the camera module 51 side. When the time lapse portion 50 is closed, the inside of the culture chambers R11 to R22 is kept in a hermetic state of a predetermined level or more by the seal portion. Be drunk.

Carbon dioxide gas (CO ₂ ) and nitrogen gas (N ₂ ) are supplied to the embryo culture apparatus 10 from an external gas cylinder. These gases are supplied, and gas ports 31 and 32 are provided on the back of the embryo culture apparatus 10. In addition, filter ports 35 and 36 to which a filter 34 is attached are also provided on the back of the embryo culture apparatus 10. The filter 34 is for removing foreign matters such as dust in the outside air taken in by a built-in pump (not shown). In the embryo culture apparatus 10, carbon dioxide gas CO ₂ and nitrogen gas N ₂ input from the gas ports 31 and 32 and air input through the filter 34 are mixed at a predetermined ratio to generate an air-fuel mixture. The ratio of each gas in the air-fuel mixture is measured by a sensor (not shown) and is always kept at the same ratio.

  FIG. 3 is a plan view showing one of the culture chambers R11 to R22. As illustrated, the culture chambers R11 to R22 are provided with a recess 40 for placing a tray (also called a dish) containing treated eggs. The shape of the tray 91 is shown in FIG. The recess 40 has a shape that matches the shape of the tray 91. In the present embodiment, the concave portion 40 has a shape in which one portion of a rectangular recess is expanded outward (expanded portion 42), and the tray 91 similarly has a shape in which one portion protrudes outward ( Projecting part 92). For this reason, the recessed part 40 is made into the shape which is not line-symmetrical with respect to a centerline even if it takes any of front and back and right and left. Accordingly, when placing the tray 91 in the recess 40, the placement direction is not mistaken. Of course, prevention of erroneous placement may be achieved by other methods, for example, electrical detection, and the shape of the recess 40 and the tray 91 may be symmetrical with respect to the left and right and / or front and rear. Further, a marker may be provided on the tray 91 so that the arrangement (orientation, etc.) of the tray 91 or the treatment egg is directly specified from the captured image regardless of the placement direction.

  A supply port 43 and an exhaust port 44 are provided on the bottom surfaces of the culture chambers R11 to R22. The supply port 43 is an opening for supplying the air-fuel mixture to the culture chambers R11 to R22. The exhaust port 44 is an opening for recirculating the air-fuel mixture. As described above, since the culture chambers R11 to R22 are almost airtight when the time lapse unit 50 is closed, the internal gas environment is kept constant, but the temperature distribution in the culture chambers R11 to R22 is kept constant. In order to achieve this, the air-fuel mixture is supplied from the supply port 43 and exhausted from the exhaust port 44. A part of the air-fuel mixture also slightly leaks from the seal part with the time lapse part 50. By doing so, outside air is prevented from entering from the seal portion with the time lapse portion 50.

  In each of the culture chambers R11 to R22, panel heaters H11 to H22 are provided on the side walls and the bottom surface. The heaters H11 to H22 may be provided only on the side wall or on the bottom surface. Alternatively, independent heaters may be provided in the culture chambers R11 to R22. Further, it may be provided on the lid side. The culture chambers R11 to R22 are kept at a constant temperature by the panel heaters H11 to H22. Although not described, the culture chambers R11 to R22 are provided with a temperature sensor, a gas concentration sensor, and a humidity sensor (not shown), and the temperature, gas concentration, and humidity in the culture chambers R11 to R22 are determined. Can be detected. By feeding back the signals from these sensors, as a result, the culture chambers R11 to R22 are kept at a constant temperature and a constant humidity, and the gas concentration is also kept constant. Instead of detecting the temperature in the culture chambers R11 to R22, the environment in the culture chambers R11 to R22 is maintained by keeping the temperature, humidity, gas concentration, etc. of the air-fuel mixture supplied from the supply port 43 constant. It may be kept constant.

  Next, the time lapse unit 50 will be described. FIG. 4 is an explanatory diagram showing an electrical configuration of the embryo culture apparatus 10 including the time lapse unit 50. As illustrated, the embryo culture apparatus 10 is provided with a control unit 60. The control unit 60 includes a well-known CPU 61, ROM 62, and RAM 63, a memory interface 64 for exchanging data with the memory card 65, a general-purpose I / O interface 66 for exchanging signals with external devices, and a culture room R11. The culture room interface 67 for exchanging signals for controlling the environment in the R22, the SIO 68 for serial communication with the time lapse unit 50, and the like are provided. In the memory card 65, information related to the environment such as the temperatures of the culture chambers R11 to R22 is recorded.

  The general-purpose I / O interface 66 includes switches SW11 to SW22 provided in the embryo culture apparatus 10, a drive device 71 that opens and closes the time lapse unit 50, a warning device 72 that generates a warning sound, and a mixture adjustment that adjusts the mixture. It is connected to the device 73 and the like. The air-fuel mixture adjusting device 73 includes a pressure adjusting valve that adjusts the pressure of carbon dioxide gas and nitrogen gas supplied to the gas ports 31 and 32, a pump for taking in air through the filter 34, and the culture chambers R11 to R11. Includes a pump that feeds the air-fuel mixture to R22. The culture chamber interface 67 is connected to panel heaters H11 to H22 provided in the culture chambers R11 to R22, control valves V11 to V22 for controlling the supply amount of the air-fuel mixture, and the like. The control valves V11 to V22 are provided in a pipe from the mixture supply pipe 84 for supplying the mixture to the culture chambers R11 to R22.

  As described above, the camera module 51 of the time lapse unit 50 includes the four camera units CA1 to CA4. These camera units CA1 to CA4 image the well 95 provided with the tray 91, as shown in FIGS. As shown in the drawing, the wells 95 in the tray 91 are arranged in groups of four at four locations. The camera units CA1 to CA4 are provided corresponding to each group. Areas U1 to U4 illustrated in FIG. 5 indicate imaging areas of the camera units CA1 to CA4, respectively. That is, each of the camera units CA1 to CA4 can capture an area including the four wells 95 at a time. A light guide for guiding light from an LED (not shown) is provided around the lenses of the camera units CA1 to CA4. The camera units CA1 to CA4 irradiate light from the light guide toward the tray 91 during imaging.

  The state of imaging is schematically shown in FIG. The well 95 is provided at the bottom of the tray 91, and one treatment egg 25 is basically placed in one well 95 and covered with mineral oil 93. Each camera unit CA <b> 1 to CA <b> 4 images areas U <b> 1 to U <b> 4 including four wells 95 from above the tray 91. An image of the treatment egg 25 is included in the captured image. The camera units CA1 to CA4 can adjust the focal position at the time of imaging with an accuracy of several tens of μm. A state of imaging the treated egg 25 by the camera units CA1 to CA4 will be described later.

  Next, the internal configuration of the time lapse unit 50 will be described. FIG. 7 is an explanatory diagram showing an internal configuration of the time lapse unit 50. As shown in the figure, the time lapse unit 50 includes a known CPU 53, ROM 54, RAM 55, digital signal processor (DSP) 56, SIO 57, memory interface 58, camera interface 81, in addition to the camera module 51 and display module 52 described above. A display interface 82 and the like are provided. In the figure, “interface” is abbreviated as “I / F”.

  A memory card 59 can be attached to the memory interface 58. As will be described later, an image picked up by the camera module 51 is recorded on the memory card 59. The camera interface 81 is connected to the camera module 51 and performs processing such as exchange of signals for controlling the focal positions of the camera units CA1 to CA4 of the camera module 51 and reception of signals of images captured by the camera units CA1 to CA4. Do. The display interface 82 is connected to the display module 52 and outputs an image signal to be displayed on the display module 52 and inputs a signal from a touch panel provided on the display module 52.

  On the premise of the hardware configuration of the first embodiment described above, the embryo culture apparatus 10 performs the following processing. In the present embodiment, the control unit 60 of the embryo culturing apparatus 10 controls the whole, and the CPU 53 of the time lapse unit 50 controls imaging and display of the treated egg 25. First, the entire process will be described with reference to FIG. FIG. 8 is a flowchart showing a culture unit control routine. This process is executed by the control unit 60.

  The culture unit control routine shown in FIG. 8 is repeatedly executed at predetermined intervals when the embryo culture apparatus 10 is powered on. When this processing routine is started, first, which culture unit is to be processed is determined (step S200). Actually, the culture unit to be processed is specified by sequentially incrementing the variable n for specifying the culture units 11 to 22 to be processed between the numerical values 11 to 22. Note that the variable n is sequentially incremented, and when the value 22 is reached, the variable n is set to the value 11 in the next increment. That is, each culture unit 11-22 becomes the object of processing at a rate of once every 12 times. When the number n of the culture unit to be processed is determined in step S200, it is next determined whether or not the flag Fn for the culture unit n is 1 (step S205). The flag Fn is reset to a value of 0 when the embryo culture device 10 is turned on.

  Therefore, before the use of each of the culture units 11 to 22 is started, it is determined that the value of the flag Fn is not a value 1, and next display before use is performed (step S210). The display before use is a process for sending a signal to the time lapse unit 50 of the culture unit n to be processed via the SIO 68 to display that the culture unit n is not in use. An example of the display is shown in FIG. Therefore, on the display module 52 of the time lapse unit 50 of the culture unit before use, all the explanatory text “Please tap“ Start ”to start using” ”is displayed, and“ Start ”is displayed on the touch screen. "Button 76 is displayed.

  Next, it is determined whether the “start” button 76 on the screen has been tapped (step S225). If the “start” button 76 has been tapped, it is next determined whether the lid has been opened or closed (step S230). The lid, that is, the time lapse unit 50 is opened and closed when the switch SWn corresponding to each culture unit n to be used is operated. By operating the switch SWn, the time lapse unit 50 of the culture unit n to be used is opened, the tray 91 containing the treated eggs 25 in the wells 95 is placed in the culture chamber Rn, and the switch SWn is operated again to operate the time lapse unit. Close 50. Thus, it is determined that the lid has been opened and closed.

  When the lid is opened and closed, it is determined that the tray 91 is disposed in the culture chamber Rn, and after executing the start process (step S240), the flag Fn is set to 1 (step S255). “NEXT” is exited and this processing routine is terminated. Here, the start process (step S240) means a process for instructing the time lapse unit 50 to start the time lapse process after setting the temperature, gas concentration, and the like in the culture chamber Rn to a desired state. The flag Fn having a value of 1 indicates that the culture room Rn is in use. When the start process is executed, information in the culture chamber is displayed on the display module 52. An example of this is shown in the column (B) of FIG. In this example, “Culture and time-lapse processing is started” is displayed, and information on the current culture chamber Rn is displayed below. Specifically, information related to imaging such as imaging start time and elapsed time, imaging interval, environment in the culture chamber, that is, information on temperature and humidity, and information such as gas concentration are displayed. The temperature and gas concentration are detected by a sensor (not shown). In addition to such information, information for identifying the patient of the treated egg, such as a medical record number, may be displayed.

  Thus, when the culture is started and the flag Fn is set to the value 1, this processing routine is started next, and at the timing when the culture chamber Rn is specified in step S200, the determination in step S205 is “YES”. The process proceeds to step S260. In step S260, the time lapse unit 50 is instructed to continue the time lapse process. The time lapse process performed by the time lapse unit 50 will be described in detail later with reference to FIG. 10. In brief, each well 95 of the tray 91 in the culture chamber is imaged, and the fertilization of the treated egg 25 in the well 95 is performed. This is a process of determining the possibility of the above and displaying a necessary image on the display module 52.

  After instructing the time lapse unit 50 to continue such time lapse processing, it is determined whether or not an instruction to end use has been accepted (step S265). The instruction to end use is obtained when the embryo cultivator operates the touch panel of the display module 52. If there is no instruction to end the use, the process directly exits to “NEXT” and ends the present processing routine. If there is an instruction to end use, it is determined whether the lid has been opened or closed (step S270). After confirming that the lid has been opened and closed, an end process (step S280) is executed. The termination process is a process for terminating the temperature control in the culture chamber Rn. Thereafter, the flag Fn is set to 0 (step S295), and this processing routine is once ended.

  Next, the time lapse process will be described with reference to FIG. FIG. 10 is a flowchart showing a time lapse control processing routine. This process is repeatedly executed at predetermined intervals in the time lapse unit 50 that has been started in step S240 in FIG. 8 until the end process in step S280 is performed.

  When the time lapse control process shown in FIG. 10 is started, it is first determined whether or not a predetermined time has elapsed since the image of the treated egg was previously captured (step S300). Here, the predetermined time corresponds to the imaging interval shown in the column (B) of FIG. Of course, it is not limited to 10 minutes illustrated in the column (B) of FIG. If a predetermined time has elapsed since the previous imaging (step S300: “YES”), an imaging process is performed (step S310). The captured image is stored in the memory card 59 together with the image capturing time data.

  The imaging process (step S310) is performed by driving the four camera units CA1 to CA4 existing in the camera module 51 of one time lapse unit 50 simultaneously or sequentially. As shown in FIG. 5, by operating each of the camera units CA <b> 1 to CA <b> 4, the state of the treatment egg 25 accommodated in the 16 wells 95 of the tray 91 is imaged. This situation is shown in FIG. Each of the camera units CA1 to CA4 images the areas U1 to U4, and includes four wells 95 therein.

  Imaging by each of the camera units CA1 to CA4 is performed from above the tray 91. At this time, the camera units CA1 to CA4 capture five images per one well 95 while moving the focal position. This situation is shown in FIG. In FIG. 12, the lenses L1 and L2 are schematically drawn outside the camera unit CA1, but actually the lenses L1 and L2 are built in the camera unit CA1. By moving at least one of the lenses L1 and L2 back and forth along the optical axis, the focal position of the camera unit CA1 is a position GL + 1 farther from the camera unit CA1 than the center position GL0, and further far away. The position can be changed to a position GL + 2, a position GL-1 that is closer to the camera unit CA1, and a position GL-2 that is closer. Since the size of the treatment egg 25 is about 100 to 150 μm in the case of a human egg, the difference between the positions is set to approximately 20 to 25 μm. Five images are captured per such camera unit CA1 to CA4. The reason why multiple images with different focal positions are taken is that, in the treated egg 25, the presence position of the pronucleus or the like, which is a clue when determining fertilization, exists at any position of the treated egg 25 that is a substantially spherical body. I don't know.

  In one captured image, four wells 95, that is, four treatment eggs 25 are shown, but the images may be stored in the memory card 59 as they are, or divided into four and each treatment egg 25 is divided. May be saved. In the present embodiment, the captured image is recorded on the memory card 59, but may be stored in a so-called cloud via a communication line instead of the memory card 59. In this case, it is stored in a storage device (hard disk or the like) on the network set in advance.

  When imaging, you may light LED (illustration omitted) provided around the lens of camera unit CA1-CA4. In this case, it is desirable to select a wavelength of the LED that hardly affects the treated egg 25. In view of the fact that the treated egg 25 is a living body, imaging using infrared light may be performed without performing illumination with an LED or the like. When illuminating with the LED, the illumination is not limited to the configuration in which the illumination is performed from the camera units CA1 to CA4, but the illumination may be performed from the side or from below. The light source may be provided in another place and guided to a position where the treatment egg 25 is illuminated by a light guide such as an optical fiber. When illuminating from the side, utilizing the high directivity of the LED, the treatment egg 25 is irradiated with light that just slices the treatment egg 25 at the five standard positions GL0, GL ± 1, and GL ± 2. You may make it project sequentially. In this case, the camera units CA1 to CA4 may perform imaging with a single focus lens without incorporating a mechanism for changing the focal position. This is because an image of a position illuminated by the LED can be taken.

  After completing the above imaging process (step S310), it is determined whether or not it is a timing at which fertilization can be determined (step S320). The determination of fertilization means determining whether fertilization has occurred in the treated egg 25 that has been fertilized. Fertilization is established by the lapse of 36 hours at the earliest about 8 hours after the insemination treatment, on average about 22 hours, even if it is late. Since the treated egg 25 subjected to fertilization treatment is immediately accommodated in the well 95 of the tray 91 and placed in the culture units 11 to 22 after the treatment, about 6 hours have passed since the imaging by the time lapse unit 50 was started. Then, the determination in step S320 is “YES”, that is, fertilization determination is possible. Naturally, this time may be further shortened so that fertilization can be determined immediately after the tray 91 is accommodated in the culture chambers R11 to R22. In that case, the determination in step S320 is not necessary.

  If it is determined that fertilization determination is possible, then fertilization determination processing is performed (step S330). This process can be determined based on whether the image of the treated egg 25 stored in the memory card 59 is read and the two pronuclei are clearly shown in the image. Of course, this determination may be made based on changes over time, in particular, changes in the number of parts that can be recognized as pronuclei, by reading a large number of images stored in the memory card 59 so far. Furthermore, many images of fertilized treated eggs and images of treated eggs that have not been fertilized are prepared, machine-learned in advance, and images read from the memory card 59 are input to this machine-learned decision machine By doing so, it may be determined whether fertilization is possible. Such processing can be realized in real time by using the high-speed calculation function of the DSP 56 shown in FIG. Note that the captured image may be displayed on the display module 52 to allow the embryo cultivator to determine whether fertilization is possible.

  After performing the fertilization determination process (step S330), it is determined whether or not the treated egg 25 is a fertilized egg whose fertilization can be confirmed (step S340). If it can be determined that the egg is a fertilized egg, a fertilized egg number is displayed (step S350). An example of this display is shown in the column (C) of FIG. In this example, an image confirming fertilization, a number display indicating what number of well 95 the fertilized egg is in, a message 77 indicating confirmation of fertilization, and an “end” button for instructing the end of culture 78 is displayed on the display module 52. Thereafter, the process exits from “NEXT” and ends this processing routine.

  In the processing shown in FIG. 10, for convenience of explanation, the imaging processing performed at a predetermined interval and the fertilization determination processing are shown in one flowchart, but both may be performed asynchronously by separate processing routines. Absent.

  Consider the display time when imaging and fertilization determination are performed as a series of processes. The tray 91 placed in one culture chamber Rn is provided with 2 × 2 × 4 (a total of 16) wells 95 and can accommodate a maximum of 16 treated eggs. One time lapse unit 50 can simultaneously image these 16 wells 95 using four camera units CA1 to CA4. Therefore, a case is considered in which the fertilization determination and the result are displayed while the treated eggs contained in the 16 wells 95 are sequentially displayed on the display module 52. An example of the display when it is determined to be fertilized has already been shown in the column (C) of FIG. 9, but when it cannot be determined that it has been fertilized, the latest image of the treated egg is displayed at that time. Alternatively, it may not be displayed. If the imaging interval is 10 minutes, a display of about 50 seconds per well 95 can be performed. Therefore, the embryo cultivator can know the number of the well 95 containing the treated egg that has been fertilized by viewing this display. It is also possible to determine the end of the culture.

  Since the fertilization treatment is rarely performed at the same time, even if there are twelve culture units 11 to 22 in the embryo culture apparatus 10 of this embodiment, the embryo culture apparatus 10 as a whole has fertilized eggs in twelve culture units at the same time. It is not displayed. Therefore, if only the fertilized eggs that have been determined to be fertilized are displayed on the display module 52, the embryo cultivator looks at the sequentially displayed images even if there are 12 culture units 11-22. , Confirmation of fertilization, removal of the tray 91, subsequent treatment, etc. can be handled.

  The embryo culturing apparatus of the first embodiment described above is provided with the time lapse unit 50 for each of the 12 culture chambers R11 to R22, and can image the treated egg 25 accommodated in the tray 91 intermittently. For this reason, in order to image the treatment egg 25, it is not necessary to move the tray 91 or an imaging device (a camera or the like). Therefore, not only the embryo culture apparatus 10 is reduced in size but also has no movable part, so that the reliability of the embryo culture apparatus 10 can be improved. Also, no vibration or noise occurs. Furthermore, the interval at which the treated egg 25 is imaged can be shortened. In the system for moving and imaging the tray 91, for example, if it takes m minutes to move and image the tray, 12 × m minutes are required to move and image all the 12 trays 91. In other words, the interval cannot be shorter than the time 12 · m. Alternatively, a system can be considered in which a plurality of trays are mounted on a rotating disk-shaped mounting table and imaged while rotating, but even in this case, it takes a significant amount of time to rotate and image the mounting table. The tray imaging interval cannot be less than the time required for one rotation of the mounting table. On the other hand, in the embryo culture apparatus 10 of the present embodiment, the imaging interval can be shortened as much as the imaging interval of the camera units CA1 to CA4. Of course, continuous imaging is also possible.

  Furthermore, according to the present embodiment, one time lapse unit 50 is prepared for one tray 91 accommodated in one culture chamber R11 to R22, and the display module 52 is provided in the time lapse unit 50. The image of the treated egg 25 of the tray 91 accommodated in the culture chambers R11 to R22 is displayed here. Therefore, there is a one-to-one relationship between the image displayed on the display module 52 and the tray 91 where the time lapse unit 50 is opened by operating the switch SWn, and the relationship between the image and the treated egg is very clear. It also has the advantage of being.

  Furthermore, in the present embodiment, the time lapse control process is performed for each time lapse unit 50. Therefore, for each time lapse unit 50 of the culture units 11 to 22, a time lapse process, for example, an imaging interval, a form of an image to be displayed, etc. Can be set individually. For example, if there are treated eggs that are particularly worried about the progress after fertilization treatment, it may be necessary to shorten the imaging interval or to ask the embryo culturer to judge when fertilization is possible. Easy.

  In addition, each of the time lapse units 50 of the present embodiment includes a memory card 59 and records images captured at predetermined intervals. Therefore, an image in which a treated egg cultured in the culture room Rn is fertilized is not mistaken for an image of another treated egg. If necessary, the captured image can be managed, stored, discarded, etc. for each memory card, and is easy to handle.

B. Second embodiment:
Next, a second embodiment of the present invention will be described. The appearance shape of the embryo culture apparatus 10A of the second embodiment is the same as that of the first embodiment. As shown in FIG. 13, the embryo culture apparatus 10 of the second embodiment has a different configuration of the control unit 60 </ b> A and a different connection relationship between the control unit 60 </ b> A and the time lapse unit 50. To briefly explain the difference, in the first embodiment, the time lapse control process is executed by the time lapse unit 50 alone, whereas in the second embodiment, the entire process including the time lapse control process is performed. This is implemented by the control unit 60A.

  FIG. 13 is an explanatory diagram showing a circuit configuration of an embryo culture apparatus 10A according to the second embodiment. As shown in the drawing, this embryo culture apparatus 10A is different from the culture room interface 67 and the SIO 68 in the control unit 60A, except that a camera interface 68A and a display interface 68B are provided. 10 and the same configuration. In FIG. 13, although not shown, panel heaters H11 to H22 and control valves V11 to V22 are provided in the culture chambers R11 to R22, and the temperature and gas concentration in the culture chambers R11 to R22 are controlled. The point is also the same.

  In the second embodiment, the camera modules 51 of the time lapse unit 50 are all connected to the camera interface 68A for each of the camera units CA1 to CA4. Therefore, the CPU 61 of the control unit 60A can individually control the camera units CA1 to CA4 of each time lapse unit 50 via the camera interface 68A to perform imaging. The display modules 52 of the time lapse unit 50 are each connected to the display interface 68B. Therefore, the CPU 61 of the control unit 60A can individually display on the display module 52 of the time lapse unit 50 provided in each of the culture units 11 to 22 via the display interface 68B.

  In the above configuration, the control unit 60A executes the process shown in FIG. This process is repeatedly executed at predetermined intervals immediately after the power is supplied to the embryo culture apparatus 10A. When this processing routine is started, processing for designating the culture unit n is first performed (step S400). The designation of the culture unit n is normally performed cyclically from the culture unit 11 to the culture unit 22. If there is a culture unit in failure, the number is stored, and designation of the culture unit in failure is avoided in the process of step S400.

  Next, the culture unit n determines whether to start use, in use, or to end use (step S405). For the culture units that are not used, the control unit 60A performs a display for specifying the start of use as shown in the column (A) of FIG. 9 via the display interface 68B. When the embryo culturer operates the touch panel of the display module 52 and taps the “start” button, the determination in step S405 is “start”, and then the process of storing the number n of the culture unit to start using. Is performed (step S410). The number n of the culture unit to be used may be stored in the RAM 63 or may be stored in the memory card 59.

  Subsequently, the temperature and gas concentration are controlled for the designated culture unit n (step S420). This process is the same as the process described in the first embodiment (FIG. 8, step S240). After starting control of temperature, humidity, and gas concentration, it is determined whether the environment of the culture chamber Rn is stable (step S425). This determination is made by detecting temperature, humidity, gas concentration and the like by a sensor (not shown). If the detected temperature, humidity, and gas concentration are all within ± 5% of the target value, it is determined to be stable.

  If it is determined that the environment of the culture room Rn is stable, the control unit 60A sends a signal to the display module 52 of the time lapse unit 50 corresponding to the culture room Rn via the display interface 68B to instruct the display of “available”. (Step S430). The “usable” display is the same display as the column (A) in FIG. 9 described in the first embodiment. When the embryo cultivator who is the user prepares the tray 91 in which the treated egg 25 is accommodated in the well 95, the “start” button displayed on the display module 52 is tapped. When the “Start” button 76 is tapped, the time lapse unit 50 is opened. This is because the time lapse part 50 functions as a lid for the culture chamber Rn.

  Thereafter, the tray 91 is accommodated in the culture chamber Rn, and the time lapse part 50 as a lid is closed. The control unit 60A waits until it is detected (step S435). When it is confirmed that the time lapse unit 50 as a lid has been opened and closed (step S435: “YES”), the time lapse process is executed next (step S440). This time lapse process is the same process as the time lapse process (FIG. 8, step S290) described in the first embodiment. During this time, the display in the column (B) of FIG. 9 is performed on the display module 52 of the time lapse unit 50. After the time lapse process (step S440) is executed, “NEXT” is exited, and this processing routine is temporarily ended.

  When the routine shown in FIG. 14 is repeatedly executed, if it is determined in step S405 that the culture unit is “in use”, the process proceeds to step S440, and a time lapse process is executed. When it is determined that the treated egg 25 has been fertilized while the time lapse process (step S440) is continuously executed after the start of use in this way, the display in the column (C) of FIG. 9 is made.

  When the main processing routine shown in FIG. 14 is repeatedly executed, fertilization of the treated egg is determined, and when the embryo cultivator taps the “end” button 78 displayed on the display module 52, the culture is performed. The process for the unit is determined to be an “end” process (step S405), and the process proceeds to step S455. In step S455, it is determined whether the time lapse unit 50 functioning as a lid has been opened or closed. When it is determined that the time lapse unit 50 functioning as a lid for the culture chamber Rn has been opened and closed, it is determined that the tray 91 has been removed from the culture chamber Rn, and the use of the culture chamber Rn has been completed. The number “n” of the culture chamber Rn is deleted from the list of the culture chambers.

  Subsequently, the control of the temperature, gas concentration and the like for the culture chamber Rn is terminated (step S470). Of course, the processing in step 470 may be continued. Thereafter, the display module 52 of the time lapse unit 50 corresponding to the culture room Rn is instructed to display “end of use”, and the process goes to “NEXT” to display the culture room. This processing routine for Rn ends.

  According to the second embodiment described above, there are advantages that the same operation and effect as in the first embodiment can be obtained, and further, the processing can be collectively performed by the control unit 60A. This is because each time lapse unit 50 operates independently in the first embodiment, but in the second embodiment, the time lapse unit 50 performs all determinations and operations such as fertilization determination by the control unit 60A. For this reason, the structure of the time lapse part 50 can be simplified.

C. Other embodiments:
In addition to the first and second embodiments described above, the present invention can be implemented as the following embodiments. In the embodiment described above, it is determined whether or not the treated egg 25 has been fertilized. However, the treated egg 25 is continuously cultured without performing such a determination or after making a determination, and the state is the imaging unit. It is good also as what is imaged with camera unit CA1-CA4. Or you may comprise as an apparatus which performs from the place which accommodates the treatment egg 25 in the well 95 of the tray 91 to the place which sends out the fertilized fertilized egg to the next treatment.

  In the above embodiment, four camera units CA1 to CA4 are prepared for each time lapse unit 50, but the number of camera units per time lapse unit 50 is arbitrary. The number of wells 95 to be imaged by the camera unit, that is, the number of treatment eggs to be imaged at a time can be 1 or more and any number. Further, the arrangement of wells in the tray 91 is also arbitrary. For example, it may be arranged in an array such as p × q (p and q are integers of 1 or more), or may be arranged in a ring shape. As long as the required resolution is obtained, it is possible to image as many treated eggs as possible. Alternatively, one camera unit may be moved within the time lapse unit 50 to capture images of all wells.

  In the above embodiment, one tray 91 is arranged in one culture chamber Rn, but two or more trays may be arranged in one culture chamber Rn. In that case, one time lapse unit 50 may be associated with a plurality of trays, or one time lapse unit 50 may be associated with one tray.

  In the above embodiment, the time lapse unit 50 and the camera module 51 are arranged on the upper part of the culture chamber Rn so as to serve as a lid. However, the lid of the culture chamber Rn may be provided separately from the camera module 51. The camera module 51 can also be disposed on the bottom surface or side surface of the culture chamber Rn. In the case of arranging on the bottom surface, the bottom surface of the culture chamber Rn is formed of a material such as transparent glass, and the camera unit CA1 and the like are disposed under the bottom surface so that the treated egg in the well 95 of the tray 91 is imaged. That's fine. In general, the treated egg sinks to the bottom of the well 95, and therefore, if the image is taken from below the floor surface, it is ready to focus the camera unit on the treated egg. In this case, only the display module 52 may be disposed as a lid on the upper part of the culture chamber Rn, or the display module 52 may be provided in another place.

  The camera module 51 may be provided on the side surface of the culture chamber Rn. In this case, the wells of the tray are arranged in a straight line. FIG. 15 shows the relationship between the trays 91A and 91B and the culture chamber Rn when the camera module 51 is provided on the side surface. As shown in the figure, eight wells 95 are arranged on a straight line in each of the trays 91A and 91B. Four treatment eggs 25 in the eight wells 95 arranged in a row on the tray 91A are imaged four by two from the two camera units CA1 and CA23 of the camera module 51A provided on the left side of the culture chamber Rn. The remaining two camera units CA2 and CA4 of the camera module 51A are provided on the right side of the culture chamber Rn, and four treated eggs on the tray 91B are imaged four by four. A state of the tray 91A viewed from the camera unit CA1 is schematically shown in FIG. A well 95 is provided on the bottom surface of the tray 91A, and the treated egg 25 is accommodated here. Therefore, the positional relationship of the treatment egg 25 with respect to the camera units CA1 to CA4 becomes stable, and the image can be easily captured.

  In this example, since the width of the trays 91A and 91B arranged in one culture chamber Rn can be reduced, a large number of culture chambers Rn and trays 91A and 91B can be arranged in one embryo culture apparatus 10. If the width is narrowed, the trays 91A and 91B and the camera unit are arranged close to each other. However, if a wide-angle lens is used, the images can be clearly captured even if they are brought close to the trays 91A and 91B. Note that the camera module 51 may be provided with camera units on the front and rear sides instead of the left and right sides or together with the left and right sides. Of course, it is good also as what is provided in one side of right and left and front and back. Alternatively, it may be provided together with the top surface and the bottom surface, and the treated egg may be imaged from a plurality of directions.

  In the above embodiment, when one treatment egg is imaged, a plurality of images are picked up by shifting the focus position. However, the focus position is not particularly shifted, and one treatment egg is set at a predetermined interval. Only one image may be taken. Note that imaging is not limited to visible light, and may be performed including areas such as near infrared, far infrared, and ultraviolet light. Alternatively, a filter that transmits only a predetermined wavelength range in visible light may be provided, and imaging may be performed only with light that passes through the filter.

  In the above embodiment, the display module 52 is provided for each time lapse unit 50, but the display module 52 may be provided separately from the time lapse unit 50. In that case, it can be integrated into a large liquid crystal panel or the like. Alternatively, it may be displayed on a display such as another computer using a wireless LAN or a wired LAN. Furthermore, an application may be put in a mobile device such as a mobile phone used by an embryo culturer and displayed here. In this way, the embryo cultivator can check the status of the treated egg in real time without being in the room where the embryo culture apparatus 10 is placed. Alternatively, it is also possible for the patient receiving the fertility treatment that provided the treated egg to check the status of the treated egg. Furthermore, in this way, a plurality of embryo cultivators, or two or more embryo cultivators and patients can confirm the status of the treated egg at the same time. Therefore, the timing of fertilization is not missed.

  In the above embodiment, the imaging timing (start time and interval) can be set for each time lapse unit 50 provided for each culture room Rn. However, the imaging start may be limited to a predetermined timing such as every hour. Good. Furthermore, the interval and the like may be common to the plurality of time lapse units 50. In addition, an instruction to start imaging is made by tapping a button displayed on the display module 52, but may be instructed by voice recognition together with other instructions. Of course, a dedicated instruction board may be prepared and a switch, a hardware button, or the like arranged on the instruction board may be operated.

  In the above embodiment, the time lapse unit 50 also serves as a lid for the culture chamber Rn, is attached to the edge of the culture chamber Rn with a hinge (not shown), and rotates as shown in FIG. Rn is opened and closed, and the tray 91 can be attached and detached. Naturally, the time lapse unit 50 may be attached to the culture chamber Rn by a method other than rotational movement. For example, the time lapse unit 50 may be simply placed on the upper surface of the culture chamber Rn. Or you may make it move parallel up and down. It is also possible to have a configuration that slides left and right or back and forth.

  In the above embodiment, the time lapse unit 50 is designed and attached as a part of the embryo culture device 10 from the beginning, but may be replaced with a lid of the culture chamber of the embryo culture device already sold and installed. . In this way, the existing embryo culture device can be easily replaced with a time-lapse type embryo culture device.

  Although various embodiments of the present invention have been described above, the present invention is not limited to these embodiments, and can of course be implemented in various forms without departing from the gist of the present invention. For example, a minimum unit in which the culture room Rn and the time lapse part 50 are paired is made, and this can be connected in the left-right direction and / or the front-rear direction, and an embryo culture apparatus can be combined with any number of culture rooms and time-lapse parts. It may be configured. In the embryo culture apparatus of the present invention, the imaging unit is provided corresponding to each tray held in the culture unit, so that it is easy to add the culture unit by such connection.

CA1 ... Camera unit H11-H22 ... Panel heater L1, L2 ... Lens R11-R22 ... Culture chamber SW11-SW22 ... Switch 9 ... Case body 10,10A ... Embry culture apparatus 11-22 ... Culture unit 25 ... Treatment egg 31 ... Gas Port 34 ... Filter 35, 36 ... Filter port 40 ... Recess 42 ... Expansion part 43 ... Supply port 44 ... Exhaust port 50 ... Time lapse part 51, 51A ... Camera module 52 ... Display module 53 ... CPU
54 ... ROM
55 ... RAM
58 ... Memory interface 59 ... Memory card 60, 60A ... Control unit 61 ... CPU
62 ... ROM
63 ... RAM
64 ... Memory interface 65 ... Memory card 66 ... General-purpose I / O interface 67 ... Culture room interface 68A ... Camera interface 68B ... Display interface 71 ... Drive device 72 ... Warning device 73 ... Mixture adjustment device 76 ... "Start" button 77 ... Message 78 ... "End" button 81 ... Camera interface 82 ... Display interface 84 ... Mixture supply piping 91, 91A, 91B ... Tray 92 ... Projection 93 ... Mineral oil 95 ... Well

Claims (11)

An embryo culture apparatus for storing a plurality of trays for storing fertilized treated eggs and holding them in a culture environment,
A culture unit for holding the plurality of trays in the culture environment;
A plurality of imaging units, provided corresponding to each of the trays held in the culture unit, and imaging units that continuously or intermittently image the existing portion of the tray;
A storage unit that stores, for each tray, an image captured by each imaging unit;
An embryo culturing apparatus comprising: a display unit that displays the stored image for each tray. The embryo culture apparatus according to claim 1,
The tray includes a plurality of recesses for holding treated eggs,
The imaging unit is an embryo culture apparatus that images at least a part of the plurality of recesses at a time.   The embryo culture apparatus according to claim 1, wherein the culture unit includes a plurality of culture chambers divided for each tray. An embryo culture apparatus according to claim 3,
The imaging unit is an embryo culture apparatus provided on at least one of an upper surface, a side surface, and a bottom surface of the culture chamber so as to face the existing portion of the tray. The embryo culture apparatus according to claim 4,
The imaging unit includes an imaging depth-of-focus changing unit that captures a plurality of images with different focus positions in the imaging,
The said memory | storage part memorize | stores the several image from which the position of the said focus differs as a set of images.   The said culture | cultivation apparatus is an embryo culture apparatus as described in any one of Claims 1-5 provided with the separate display part provided for every said several tray corresponding to the said imaging part.   The embryo culture apparatus according to claim 6, wherein the individual display unit is provided on an upper surface of an openable / closable lid provided in the culture unit.   The embryo culture apparatus according to any one of claims 1 to 7, further comprising: a light guide that guides light from a light source to illuminate at least a portion where the treatment egg exists in each of the plurality of trays. The embryo culture apparatus according to any one of claims 1 to 8,
A fertilization determining unit that determines whether at least one of the treated eggs in the tray has been fertilized based on an image captured by the imaging unit;
Provided corresponding to the plurality of trays that can be held in the culture unit, and when the fertilization determining unit determines that at least one of the treated eggs has been fertilized, the treated egg determined to have been fertilized is contained. An informing unit for informing a specific tray;
Embryo culture apparatus equipped with. The embryo culture apparatus according to any one of claims 1 to 9,
An instruction unit for individually instructing start of imaging to the plurality of imaging units,
Each of the plurality of imaging units receives the instruction from the instruction unit and performs the imaging at an independent timing. An imaging device that is attached to a culture unit of an embryo culture device that contains a plurality of trays that contain fertilized treatment eggs and that is maintained in a culture environment,
Corresponding to each of the plurality of trays held in the culture unit, a plurality of imaging units for imaging the existing portion of the tray continuously or intermittently;
A storage unit that stores, for each tray, an image captured by each imaging unit;
An imaging apparatus for an embryo culture apparatus, comprising: a display unit that displays the stored image for each tray.

Images