WO2018070288A1 - Système de détermination de fécondation - Google Patents

Système de détermination de fécondation Download PDF

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Publication number
WO2018070288A1
WO2018070288A1 PCT/JP2017/035857 JP2017035857W WO2018070288A1 WO 2018070288 A1 WO2018070288 A1 WO 2018070288A1 JP 2017035857 W JP2017035857 W JP 2017035857W WO 2018070288 A1 WO2018070288 A1 WO 2018070288A1
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Prior art keywords
egg
determination
image
fertilized
unit
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PCT/JP2017/035857
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English (en)
Japanese (ja)
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憲隆 福永
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憲隆 福永
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T1/00—General purpose image data processing
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00—Image analysis

Definitions

  • the present invention relates to a fertilization determination system.
  • selection of embryos is performed by evaluating the embryos in culture from the viewpoint of improvement in conception rate and QOL of patients.
  • a method for non-invasively evaluating an embryo there is a method for evaluating the developmental state of an embryo through morphological observation with a microscope.
  • the present invention has been made to solve at least a part of the problems described above, and can be realized as the following forms.
  • a fertilization determination system includes a culture unit that cultures a fertilized egg, and an image acquisition unit that captures a state of the egg cultured by the culture unit at set time intervals and acquires a plurality of images. And a determination unit that determines whether the egg is fertilized by analyzing the state of the egg reflected in the image. If it is set as such an aspect, the determination part can determine whether the egg is fertilizing based on the analysis of the image by which the state of the fertilized egg was image
  • the determination unit determines whether the egg is fertilized by analyzing changes over time of the egg in at least two images acquired at different times. May be. If it is set as such an aspect, the time-dependent change of an egg can be analyzed by analyzing at least 2 image. For this reason, it can be determined whether the egg is fertilized based on the change of the egg over time.
  • the determination unit may learn about the characteristics of the egg shown in the image by supervised learning and determine whether the egg is fertilized based on the learning. . With such an aspect, it can be determined whether the egg is fertilized based on the criterion learned by the determination unit through supervised learning.
  • the determination unit learns about the characteristics of the egg shown in the image by deep learning using a multilayer neural network, and the egg is fertilized based on the learning. It may be determined. If it is set as such an aspect, it can be determined whether the egg is fertilizing based on the reference
  • the determination unit may include a notification unit that notifies a determination image used for determination among the images. If it is set as such an aspect, the user who uses a fertilization determination system can know which image the determination part determined fertilization.
  • the notification unit may notify the determination image and may notify the determination image in the determination image based on the determination basis.
  • the user using the fertilization determination system can know which image the determination unit uses to determine fertilization, and the determination unit can know the information on which the determination is based. it can.
  • a method for selecting a non-fertilized egg is provided.
  • This method of selecting non-fertilized eggs is a method of selecting non-fertilized eggs that have not been normally fertilized among fertilized eggs, a culture step of culturing the eggs, and a culture in the culture step.
  • a non-fertilized egg can be selected based on an analysis of an image in which a fertilized egg is photographed. For this reason, the method of selecting the non-fertilized egg which is not normally fertilized among the eggs which have been fertilized is provided.
  • the present invention can be realized in various forms other than the fertilization determination system.
  • the present invention can be realized in the form of a fertilization determination device.
  • the present invention is not limited to the above-described embodiments, and it is needless to say that the present invention can be implemented in various forms without departing from the spirit of the present invention.
  • FIG. 1 is an explanatory diagram illustrating a configuration of a fertilization determination system 10 according to the first embodiment.
  • the fertilization determination system 10 is a system that determines whether a fertilized human egg is fertilized.
  • the fertilized egg is an egg that has been subjected to fertilization treatment such as a conventional method in which sperm and an egg are co-cultured or a microinsemination method.
  • the fertilization determination system 10 includes a culture unit 110, an image acquisition unit 130, a control unit 140, a user interface 150, and a notification unit 160.
  • the culture unit 110 is a so-called incubator that cultures fertilized eggs.
  • the culture conditions such as temperature, humidity, oxygen concentration, carbon dioxide concentration, and culture time in the culture unit 110 are controlled by the control unit 140 based on the contents previously input by the user via the user interface 150.
  • the culture unit 110 cultures the eggs in a state where the culture container 200 that is a container for culturing the fertilized egg is fixed in the culture unit 110.
  • the culture vessel 200 is a well plate having 12 wells in 3 rows and 4 columns. Numbers 1 to 12, which are well numbers, are attached to the bottom of each well of the culture vessel 200.
  • the culture container 200 is a container for putting one egg in each well and culturing.
  • the culture unit 110 has a container transport unit 115.
  • the container transport unit 115 has a shape extending in the horizontal direction and constitutes a bottom surface portion of the culture unit 110.
  • the container transport part 115 has a fixing part (not shown) for fixing the culture container 200 on the surface facing the upper side in the gravity direction.
  • the culture unit 110 cultivates eggs in a state where the culture container 200 is fixed to the fixed unit.
  • the container transport unit 115 transports the culture container 200 fixed to the fixed unit by moving the fixed unit to a position below the gravitational direction of the image acquisition unit 130 that partially protrudes inside the culture unit 110. .
  • the container transport unit 115 transports the culture container 200 to the initial position. In FIG. 1, the position where the culture vessel 200 is shown is the initial position.
  • the frequency (time interval) at which the container transport unit 115 transports the culture container 200 is controlled by the control unit 140 based on the contents set in advance by the user via the user interface 150.
  • the container transport unit 115 is controlled to transport the culture container 200 to the position below the image acquisition unit 130 in the gravity direction every 15 minutes.
  • the container transport unit 115 adjusts the position of the culture container 200 two-dimensionally in the vertical and horizontal directions as viewed from the image acquisition unit 130 after the culture container 200 is transported to a position below the gravity direction of the image acquisition unit 130.
  • Each well can be arranged at a position where the image acquisition unit 130 can acquire an image.
  • the container transport unit 115 transports the culture container 200 to the initial position. The process of acquiring an image of each well in the culture vessel 200 by the image acquisition unit 130 will be described later.
  • the image acquisition unit 130 acquires images by photographing the state of eggs cultured by the culture unit 110 at set time intervals.
  • the image acquisition unit 130 captures a plurality of images for each well in the culture container 200 in a time-series manner by photographing the state of the egg each time the container transport unit 115 transports the culture container 200.
  • the image acquisition unit 130 may acquire images by capturing images at time intervals directly instructed by the control unit 140.
  • the position of the well from which the image acquisition unit 130 acquires images among the 12 wells of the culture vessel 200 is designated by the user via the user interface 150 in advance. In the following description, a well position designated by the user is referred to as a “designated position”.
  • the image acquisition unit 130 is a CCD camera.
  • the control unit 140 is constituted by a microcomputer including a central processing unit and a main storage device.
  • the control unit 140 controls each unit of the fertilization determination system 10.
  • the control unit 140 controls the culture unit 110, the container transport unit 115, and the image acquisition unit 130 based on the contents previously input by the user via the user interface 150.
  • the control unit 140 includes an image storage unit 142 and a determination unit 144.
  • the image storage unit 142 stores the image acquired by the image acquisition unit 130.
  • the image storage unit 142 sends the acquired image of each well to the determination unit 144.
  • the determination unit 144 determines whether the egg is fertilized by analyzing the state of the egg shown in the image sent from the image storage unit 142. In the present embodiment, the determination unit 144 uses the images of each well acquired by the image acquisition unit 130 at a time interval of 15 minutes for 24 hours after the culture unit 110 starts culturing eggs. Determine if is fertilized.
  • the determination unit 144 determines whether the egg is fertilized by checking the number of pronuclei in the egg. In a normally fertilized egg, two pronuclei generally appear within 22 hours after fertilization.
  • the determination unit 144 determines whether an egg is fertilized by analyzing two images acquired at different times. In the present embodiment, among the images acquired by the image acquisition unit 130, two images acquired at different times are determined as two images that can most significantly recognize the difference in the number of pronuclei in the egg. The unit 144 selects. Only when the two images are two images, one image in which the number of pronuclei in the egg is not confirmed and one image in which two pronuclei in the egg are confirmed, the determination unit 144 Determine that the egg is fertilized. As described above, since the temporal change of the eggs in the two images can be analyzed, it can be determined whether the egg is fertilized based on the temporal change of the eggs. Further, by comparing the difference in the number of pronuclei in two images, fertilization can be determined with higher accuracy than determination using one image.
  • the determination unit 144 learns about the characteristics of the egg shown in the image by supervised learning, and determines whether the egg is fertilized based on the learning. In the present embodiment, the determination unit 144 learns about the determination of the number of pronuclei in the egg shown in the image, and determines whether the egg is fertilized based on the learning. For this reason, the determination unit 144 can determine whether the egg is fertilized based on the determination criterion for the number of pronuclei learned by supervised learning.
  • FIG. 2 is an example of an image labeled as an egg in which a pronucleus has not been confirmed in the fertilized egg.
  • FIG. 3 is an example of an image labeled as an egg in which one pronucleus has been confirmed in the fertilized egg.
  • FIG. 4 is an example of an image labeled as an egg in which two pronuclei have been confirmed in the fertilized egg.
  • FIG. 5 is an example of an image that is labeled as an egg in which three pronuclei have been confirmed in a fertilized egg.
  • a fertilized egg when a pronucleus is not confirmed, it is generally determined to be a non-fertilized egg. In addition, in the fertilized egg, when one or three or more pronuclei are confirmed, it is generally determined that the egg is abnormally fertilized.
  • the user interface 150 exchanges information with the user of the fertilization determination system 10.
  • the user interface 150 is a touch panel that displays an image and receives an instruction input from the user on the image.
  • the user interface 150 may include a push button that receives an instruction input from the user.
  • reports the determination image used for determination among images.
  • the notification unit 160 notifies the determination image when the user is viewing the determination result by the determination unit 144 on the touch panel screen as the user interface 150. For this reason, the user who uses the fertilization determination system 10 can know which image the determination unit 144 has determined fertilization. In this embodiment, since the time acquired for each acquired image is attached to the image, the user can know when the egg is fertilized by checking the time attached to the determination image. it can.
  • the notification unit 160 notifies the determination image 144 of information used as a basis for determination in the determination image.
  • the notification unit 160 notifies the determination unit 144 of information that is the basis for the determination.
  • reports the positional information about the part which the determination part 144 recognized as a pronucleus in the determination image.
  • FIG. 6 is a flow showing an image acquisition process executed by the control unit 140.
  • the control unit 140 executes an image acquisition process when the culture vessel 200 is transported to a position below the image acquisition unit 130 in the direction of gravity.
  • the control unit 140 calculates a variable A representing the number of wells designated by the user (step S100). After calculating the variable A indicating the number of designated wells (step S100), the control unit 140 obtains an image of the well with the smallest number among wells that have been designated and have not been photographed. The acquisition unit 130 is caused to photograph (step S110).
  • the numbers are 1 to 12 well numbers given to the bottom of each well.
  • step S110 the image acquisition unit 130 acquires a well image in the following steps (1) and (2).
  • the control unit 140 causes the container transport unit 115 to adjust the position of the culture vessel 200 two-dimensionally, and moves the position of the well to a position where the image acquisition unit 130 can perform imaging.
  • the control unit 140 moves the well to a position where the image acquisition unit 130 can perform imaging, and then causes the image acquisition unit 130 to perform imaging.
  • control unit 140 After an image of the well with the smallest number among the designated wells that have not been shot, the control unit 140 stores the number of the well that has been shot (step S110). S120). After storing the number of the well for which imaging has been completed (step S120), the control unit 140 decrements the variable A. (Step S130).
  • step S130 the control unit 140 determines whether the variable A has become 0 (step S140). When the variable A becomes 0 (step S140: YES), the control unit 140 ends the image acquisition process of FIG.
  • step S140: NO When the variable A is not 0 (step S140: NO), the control unit 140 returns to step S110 and repeats the processing of steps S110 to S140 until the variable A becomes 0.
  • step S140: YES When the variable A becomes 0 (step S140: YES), the control unit 140 ends the image acquisition process of FIG.
  • FIG. 7 is a flow showing fertilization determination processing executed by the control unit 140.
  • the control unit 140 executes fertilization determination processing when 7 hours have elapsed since the start of egg culture. In addition, the control unit 140 repeatedly executes the fertilization determination process every hour from 7 hours to 24 hours after the start of egg culture.
  • the control unit 140 calculates a variable A representing the number of wells designated by the user (step S200). After calculating the variable A representing the number of designated wells (step S200), the control unit 140 extracts an image of the well with the smallest number among the designated wells that have not been determined. (Step S210).
  • the image of the designated well is displayed after the culture unit 110 starts the egg culture. Since it is acquired by the image acquisition unit 130 at a time interval of 15 minutes during 7 hours, there are 28 images for each designated well.
  • step S210 the control unit 140 controls 28 pieces of one of the designated wells. Extract images.
  • the control unit 140 extracts 96 images for one of the designated wells.
  • control unit 140 After extracting the image of the well with the smallest number among the designated wells that have not been determined (step S210), the control unit 140 determines the pronucleus in the egg in the extracted image. Two images that can recognize the difference in number most remarkably are selected (step S220).
  • the determination unit 144 in the control unit 140 compares the two images. It is determined whether fertilization is performed (step S230).
  • control unit 140 After comparing the two images to determine whether fertilization is performed (step S230), the control unit 140 stores the number of the well for which determination has been completed (step S240). After storing the number of the well for which the determination has been completed (step S240), the control unit 140 decrements the variable A. (Step S250).
  • step S250 the control unit 140 determines whether the variable A has become 0 (step S260). When the variable A becomes 0 (step S260: YES), the control unit 140 ends the fertilization determination process of FIG.
  • step S260 If the variable A is not 0 (step S260: NO), the control unit 140 returns to step S210 and repeats the processing of steps S210 to S260 until the variable A becomes 0. When the variable A becomes 0 (step S260: YES), the control unit 140 ends the fertilization determination process of FIG.
  • the determination unit 144 can determine whether or not the egg is fertilized based on the analysis of the image in which the state of the fertilized egg is photographed. For this reason, the system which can confirm fertilization of the fertilized egg is provided. In addition, since the fertilized eggs as specimens are mechanically analyzed, more specimens can be confirmed than humans can confirm. Moreover, since it is determined by the determination part 144 whether it is fertilizing, the complexity of determination can be reduced compared with human checking with eyes.
  • Second embodiment The configuration of the fertilization determination system in the second embodiment is the same as the configuration of the fertilization determination system 10 in the first embodiment, except that a determination unit that performs learning different from the determination unit 144 in the first embodiment is provided.
  • the determination unit in the second embodiment learns about the characteristics (number of pronuclei) of an egg shown in the image by deep learning using the deep neural network 400 that is a multilayer neural network, and the egg is fertilized based on the learning. Judge whether you are doing. For this reason, it can be determined whether the egg is fertilized based on the criterion learned by the determination unit through deep learning.
  • the feature amount of a normally fertilized egg extracted by deep learning is a feature amount that cannot be recognized by a human
  • fertilization confirmation of the egg can be performed with higher accuracy than that performed by a human.
  • FIG. 8 is an explanatory diagram explaining image learning by deep learning.
  • the deep neural network 400 is a network that models a learning mechanism in the human brain nervous system.
  • the deep neural network 400 includes an input layer 410, a plurality of intermediate layers 420, and an output layer 430.
  • the deep neural network 400 according to the second embodiment includes four intermediate layers 420.
  • the input layer 410 is a layer into which information is input.
  • the intermediate layer 420 is a layer that calculates a feature amount based on information transmitted from the input layer 410.
  • the output layer 430 is a layer that outputs a result based on information transmitted from the intermediate layer 420.
  • An image 300 in FIG. 8 is an image showing an egg that has been fertilized.
  • an image labeled as an egg in which no pronuclei have been confirmed an image labeled as an egg in which one pronucleus has been confirmed, and an egg in which two pronuclei have been confirmed.
  • An image labeled as being, an image labeled as being an egg in which three pronuclei have been confirmed, and the like are included.
  • the learning method using the deep neural network 400 will be described.
  • the input layer 410 transmits the information to the intermediate layer 420.
  • the intermediate layer 420 calculates the feature value of the pronucleus based on the information transmitted from the input layer 410.
  • the output layer 430 outputs the feature amount of the pronuclei calculated based on information transmitted from the intermediate layer.
  • the determination part in 2nd Embodiment determines whether the egg is fertilized on the basis of the output feature-value.
  • non-fertilized eggs that have not been normally fertilized among the fertilized eggs will be described.
  • This method of selecting a non-fertilized egg includes a culture process, an image acquisition process, and a selection process.
  • non-fertilized egg as used herein includes a non-fertilized egg that is an egg whose pronucleus have not been confirmed, and an abnormally fertilized egg that is an egg whose one or more pronuclei have been confirmed.
  • the culture process is a process of culturing the fertilized egg.
  • the image acquisition process is a process of acquiring a plurality of images by photographing the state of the eggs cultured in the culture process at set time intervals.
  • the selection step is a step of selecting a non-fertilized egg by analyzing the state of the egg shown in the image.
  • the third embodiment described above it is possible to select a non-fertilized egg based on an analysis of an image in which the state of the fertilized egg is photographed. For this reason, the method of selecting the non-fertilized egg which is not normally fertilized among the eggs which have been fertilized is provided.
  • the fertilization determination system 10 was provided with the container conveyance part 115,
  • the fertilization determination system in another embodiment may not include the container transport unit 115.
  • the image acquisition unit 130 may be provided on the upper side in the gravity direction of the culture vessel 200 fixed to the fixing unit, or the image acquisition unit 130 is configured to be movable on the upper side in the gravity direction of the culture vessel 200. Just do it.
  • the image acquired by the image acquisition unit 130 is stored in the image storage unit 142, but the present invention is not limited to this.
  • the image acquired by the image acquisition unit 130 may be stored on a so-called cloud.
  • the determination unit 144 acquires an image from the cloud and performs determination.
  • the determination unit 144 determines whether an egg is fertilized by analyzing two images acquired at different times, but the present invention is not limited to this.
  • the determination unit 144 may determine whether the egg is fertilized by analyzing one image in which the number of pronuclei in the egg (selected by the control unit 140) can be recognized most frequently. In this case, for example, the determination unit 144 determines that the egg is fertilized when one image selected by the control unit 140 is an image in which two pronuclei are confirmed.
  • the determination unit 144 may determine whether the egg is fertilized by analyzing three images acquired at different times (selected by the control unit 140). In this case, the determination unit 144, for example, one image in which the number of pronuclei in the egg is not confirmed, one image in which one pronucleus in the egg is confirmed, and an image in which two pronuclei in the egg are confirmed. It is determined that the egg is fertilized only when three of them are confirmed.
  • the determination unit 144 may determine whether the egg is fertilized by analyzing four or more images acquired at different times (selected by the control unit 140). In this case, for example, the determination unit 144 arranges four or more images in time series, and the egg is fertilized only when the number of pronuclei increases or decreases in the order of 0, 1, 2, and 0. It is determined that
  • the input image 300 is an image labeled with respect to the number of pronuclei, but the present invention is not limited to this.
  • the input image may be an image labeled as fertilized and an image labeled as not fertilized.
  • the present invention is not limited to the above-described embodiments, examples, and modifications, and can be realized with various configurations without departing from the spirit of the invention.
  • the technical features in the embodiments, examples, and modifications corresponding to the technical features in each embodiment described in the summary section of the invention are to solve some or all of the above-described problems, or In order to achieve part or all of the above-described effects, replacement or combination can be performed as appropriate. Further, if the technical feature is not described as essential in the present specification, it can be deleted as appropriate.

Abstract

L'invention concerne un système de détermination de fécondation comprenant : une unité de culture qui cultive un œuf qui a subi un processus de fécondation ; une unité d'acquisition d'images qui acquiert une pluralité d'images par capture, à des intervalles de temps définis, d'images de l'état de l'œuf étant cultivé par l'unité de culture ; et une unité de détermination qui détermine si l'œuf a été fécondé par analyse de l'état de l'œuf capturé sur les images.
PCT/JP2017/035857 2016-10-11 2017-10-02 Système de détermination de fécondation WO2018070288A1 (fr)

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