Intracytoplasmic sperm injection (ICSI) support device, intracytoplasmic sperm injection (ICSI) support system, intracytoplasmic sperm injection (ICSI) support program, and egg retention device

Abstract

To provide an intracytoplasmic sperm injection support device with a simple device configuration. [Solution] Sperm held in sperm injection tool 3 is held by egg holding device An intracytoplasmic sperm injection (ICSI) support device 2 that assists in intracytoplasmic sperm injection (ICSI) in which sperm is injected into an egg, comprising a sperm injection tool 3 and an egg An acquisition unit that acquires information including the relative positional relationship with the child, and the information acquired by the acquisition unit Based on this, an egg retention device is positioned to perform intracytoplasmic sperm injection (ICSI) by injecting sperm into the egg. An intracytoplasmic sperm injection support device 2 comprises a control unit that controls the movement of stage 11.

Description

Technical Field

[0001] The present invention relates to an assisted micromanipulation device, an assisted micromanipulation system, an assisted micromanipulation program, and an egg holding device.

Background Art

[0002] As one of infertility treatments, intracytoplasmic sperm injection (ICSI), which directly injects sperm into the egg cytoplasm to achieve fertilization, has been developed and put into practical use. In ICSI, operations such as sperm immobilization, sperm aspiration, and sperm injection into the egg are performed while observing sperm and eggs under a microscope. Therefore, ICSI is also called micromanipulation. Currently, although micromanipulation is performed by embryologists, since it takes a very long time to master micromanipulation, technologies for assisting micromanipulation are in demand. For example, Patent Document 1 discloses a control device for moving a surgical instrument manually operated by an embryologist to a desired position.

[0003]

[0004]

Prior Art Documents

Patent Documents

Patent Document 1

[0005]

Summary of the Invention

Problems to be Solved by the Invention

[0006] Support devices, intracytoplasmic sperm injection support systems, intracytoplasmic sperm injection support programs, or egg retention devices, etc. The purpose is to provide it. [Means for solving the problem]

[0006] One aspect of this disclosure relates to an intracytoplasmic sperm injection support device that holds sperm in a sperm injection tool and egg storage This is an intracytoplasmic sperm injection (ICSI) support device that assists in intracytoplasmic sperm injection by injecting eggs held by a device. This involves an acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg, and the acquisition Based on the information obtained by the department, egg storage is performed in order to inject sperm into eggs for intracytoplasmic sperm injection (ICSI). It includes a control unit that controls the movement of the stage on which the device is placed.

[0007] Such intracytoplasmic sperm injection support devices include the relative positional relationship between the acquired sperm injection tool and the egg. Based on the information, the stage is moved to support intracytoplasmic sperm injection (ICSI). Therefore, even if Unlike devices that control the movement of sperm injection tools to support intracytoplasmic sperm injection, such as those described in Patent Document 1, Furthermore, the intracytoplasmic sperm injection system has a configuration that omits the mechanism for controlling the movement of the sperm injection tool. It is possible.

[0008] An egg retention device relating to one aspect of this disclosure includes a recess for containing an egg, and a recess for storing the egg in the recess. The device comprises a suction port for holding the egg in the above-mentioned intracytoplasmic sperm injection (ICSI). It is used in conjunction with precision support devices.

[0009] According to such an egg retention device, the intracytoplasmic sperm injection support device fixes and rotates the egg contained within. Because it can be done, for example, without controlling the movement of the sperm injection tool, The sperm injection tool can be inserted into the egg from that direction.

[0010] The intracytoplasmic sperm injection support system according to one aspect of the present disclosure includes an intracytoplasmic sperm injection support device and a sperm injection tool.

[0011] The intracytoplasmic sperm injection support program according to one aspect of the present disclosure is an intracytoplasmic sperm injection support program that supports intracytoplasmic sperm injection for injecting sperm held by a sperm injection tool into an egg held by an egg holding device, and causes a computer to function as an acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg, and a control unit that controls the movement of a stage on which the egg holding device is disposed in order to perform intracytoplasmic sperm injection by injecting sperm into the egg based on the information acquired by the acquisition unit.

[0012] Such an intracytoplasmic sperm injection support system and intracytoplasmic sperm injection support program support intracytoplasmic sperm injection by moving the stage based on information including the relative positional relationship between the acquired sperm injection tool and the egg. For this reason, for example, unlike a device that supports intracytoplasmic sperm injection by controlling the movement of a sperm injection tool as in Patent Document 1, the intracytoplasmic sperm injection system can have a configuration in which a mechanism for controlling the movement of the sperm injection tool is omitted.

[0013] Note that the program according to one aspect of the present disclosure can be installed or loaded into a computer and / or a device by being downloaded through various recording media such as optical disks such as CD-ROMs, magnetic disks, semiconductor memories, etc., or via a communication network or the like.

[0014] In addition, in this specification and the like, the term "part" does not simply mean a physical configuration, but ​​​​​​​​This includes cases where the functions of the configuration are realized by software. Also, even if the functions of one configuration are realized by two or more physical configurations, or the functions of two or more configurations are realized by one physical configuration, it is acceptable. Even if the functions of one configuration are realized by two or more physical configurations, or the functions of two or more configurations are realized by one physical configuration, it is acceptable.

Advantages of the Invention

[0015] According to the present invention, it is possible to provide a microinsemination support device, a microinsemination support system, a microinsemination support program, or an egg holding device having a simple device configuration.

Brief Description of the Drawings

[0016] [Figure 1] It is an overall conceptual diagram of the microinsemination support system according to the present embodiment. [Figure 2] It is an example of a functional block diagram of the microinsemination support device according to the present embodiment. [Figure 3] It is an example of the physical configuration of the microinsemination support device according to the present embodiment. [Figure 4] It is a schematic perspective view (A) and an enlarged plan view (B) of the egg holding device according to the present embodiment. [Figure 5] It is a flowchart showing an example of the steps of the microinsemination method according to the present embodiment. [Figure 6] It is a schematic perspective view of another example of the egg holding device according to the present embodiment.

Modes for Carrying Out the Invention

[0017] Hereinafter, modes for carrying out the present invention (hereinafter referred to as "the present embodiment") will be described in detail with reference to the drawings. However, the present invention is not limited thereto, and various modifications are possible without departing from the gist thereof. In the following description of the drawings, the same or similar parts are denoted by the same or similar reference numerals. The drawings are schematic and not necessarily actual. Although it will be described in detail, the present invention is not limited thereto, and various modifications are possible without departing from the gist thereof. In the following description of the drawings, the same or similar parts are denoted by the same or similar reference numerals. The drawings are schematic and not necessarily actual. In the description of the following drawings, the same or similar parts are denoted by the same or similar reference numerals. The drawings are schematic and not necessarily actual. In the description of the following drawings, the same or similar parts are denoted by the same or similar reference numerals. The drawings are schematic and not necessarily actual. ​Dimensions and proportions may not match. Even between drawings, the relationships between dimensions and proportions may differ. It may contain minutes.

[0018] [Intracytoplasmic sperm injection support system] Referring to Figure 1, the intracytoplasmic sperm injection support system 1 according to this embodiment will be described. Figure 1 is an overall conceptual diagram of the intracytoplasmic sperm injection support system 1 according to this embodiment.

[0019] The intracytoplasmic sperm injection support system 1 according to this embodiment is an intracytoplasmic sperm injection support device 2 according to this embodiment and The device comprises a sperm injection tool 3 and a microinsemination support device 2 according to this embodiment. This will be explained later.

[0020] As shown in Figure 1, the intracytoplasmic sperm injection support device 2 moves the sperm injection tool 3 and the stage 11. The unit consists of a drive unit 12 for performing the operation and an imaging unit 13 for imaging the sample placed on the stage 11. They are connected. Note that in Figure 1, the intracytoplasmic sperm injection support device 2 is connected to the other components by wires. However, the intracytoplasmic sperm injection support device 2 may be wirelessly connected to other components, or other components It may be physically integrated with the constituent elements. Therefore, the stage 11, the drive unit 12, and One or more of the imaging units 13 are included in the intracytoplasmic sperm injection support device 2 as part of the intracytoplasmic sperm injection support device 2. It's okay if it's not allowed.

[0021] Stage 11 is where a container 14 containing the sample is placed. The container 14 is made of, for example, glass. The dish is made of stainless steel or resin. In Figure 1, container 14 contains a sperm pool PS and eggs. A sperm pool PO is provided. Container 14 and / or stage 11 contain a sperm pool PS. and / or a temperature control means for maintaining the oocyte pool PO at a temperature suitable for intracytoplasmic sperm injection is provided. It may be. Examples of temperature control means include heaters and coolers. The control means may have a function to measure the temperature inside the container 14. A temperature suitable for intracytoplasmic sperm injection (ICSI). For example, the temperature range is around 37°C, and also 33-40°C, 35-38°C, 36-37°C, Alternatively, the temperature may be 37°C.

[0022] Sperm pool PS is a collection of sperm obtained from patients or subjects who wish to undergo intracytoplasmic sperm injection (ICSI). Includes. Sperm Pool PS contains sperm, sperm culture medium, or a highly viscous liquid that inhibits sperm motility. It may contain a body (for example, polyvinylpyrrolidone (PVP) solution), etc. Also, oocytes Eggs obtained from patients or subjects who wish to undergo intracytoplasmic sperm injection (ICSI) include eggs.

[0023] The egg pool (PO) may contain eggs, as well as egg culture medium, etc. This is not particularly limited to physiological saline (containing 0.9 w / v% sodium chloride). Saltwater can be used as an example. Oocyte pools (PO) can hold eggs using an oocyte retention device. The device is provided on or adjacent to the egg retention device.

[0024] Stage 11 is equipped with a drive unit 12 for moving the stage. The drive unit 12 is The stage 11 can be moved in the in-plane direction (x and y directions in Figure 1). However, even in the direction perpendicular to the surface of stage 11 (the z-direction in Figure 1), the stage 11 is also It is preferable that it be movable. That is, the stage 11 is equipped with a drive unit 12. It is preferable that the stage is capable of movement in the axial direction. The drive unit 12 is centered on each axis x, y, and z. The stage may be rotated in this manner. At this time, the drive unit 12 controls the x, y, and z axes. It is possible to control movement with a total of 6 degrees of freedom, including translational and rotational movement.

[0025] The drive unit 12 can move the stage 11 under the control of the intracytoplasmic sperm injection support device 2. There are no particular limitations, and for example, it may be an electrically controllable motor. Stage 11 and The drive unit 12 is, for example, an electrically controllable automatic stage provided in a microscope. stomach.

[0026] The imaging unit 13 captures images or moving images of the sample on the stage 11 and the sperm injection tool 3. The image is obtained. In Figure 1, the imaging unit 13 is positioned vertically above the stage 11. The sample shown above is being imaged in a plan view. The imaging unit 13 also captures at least the sperm injection tool 3. The sperm injection tool 3 is designed to acquire positional information for the part used for sperm aspiration and injection. Image acquisition is performed so that the relevant portion is included in the field of view. The imaging unit 13 is used to capture the sample on the stage 11. It is sufficient to obtain images or videos that include the positional relationship of the sperm injection tool 3. It does not necessarily have to be located vertically above Stage 11.

[0027] The imaging unit 13 is composed of an imaging device equipped with, for example, a CCD camera or a CMOS sensor. The imaging unit 13 uses an optical microscope to view the sample on the stage 11 and the sperm injection tool. Image 3 may be imaged. The observation method using an optical microscope is not particularly limited, but bright-field imaging is also possible. Observation methods, dark-field observation, phase contrast observation, polarization observation, spindle observation, differential interference observation, and Examples include the relief contrast observation method. Furthermore, the magnification of the objective lens can be adjusted appropriately. Often, the magnification can be, for example, 20x, 40x, 60x, etc. The data captured by the imaging unit 13 The image may be a still image or a moving image. In the following, the imaging unit 13 acquires a moving image. The following will explain the case, but the same applies when the imaging unit 13 acquires multiple still images. Needless to say, we can provide support for sperm production.

[0028] Sperm injection tool 3 is a tool for injecting specific sperm into the cytoplasm of an egg. Injection tool 3 is, for example, an injection pipette commonly used in intracytoplasmic sperm injection. That's good. The sperm injection tool 3 is good because it can be brought into contact with the sperm pool PS and used to aspirate sperm. It seems. The sperm injection tool 3 is equipped with means for immobilizing sperm in the sperm pool PS. It is also possible to do so. As a means of immobilizing sperm, weak vibrations such as those of a piezoelectric element may be applied. Means for injecting, and means for irradiating with laser light, etc. Alternatively, the tip of the sperm injection tool 3. The sperm may be immobilized by applying direct pressure to them using (for example, an inlet or outlet). The means of activation is a mechanism different from that of sperm injection tool 3, and is an intracytoplasmic sperm injection support system. It is fine to have it in Mu1.

[0029] The sperm injection tool 3 may be equipped with a pressure control mechanism for aspirating and injecting sperm. The pressure control mechanism is based on the control of the intracytoplasmic sperm injection support device 2, and the sperm injection tool 3 also has This adjusts the output of the pump attached to the sperm injection tool 3, and the inlet and outlet of the sperm injection tool 3. It sucks up sperm present in the vicinity, and inserts and removes sperm that have been sucked up and held within the sperm injection tool 3. You can spit it out orally.

[0030] The sperm injection tool 3 is preferably positioned so that its relative position to the imaging unit 13 is fixed. In this configuration, even if the stage 11 is moved, the imaging view of the imaging unit 13 In the field, the position of sperm injection tool 3 does not move, so the sample on stage 11 and sperm injection The relative positional relationship with the input tool 3 can be easily adjusted.

[0031] The intracytoplasmic sperm injection support device 2 acquires the sample and sperm injection on the stage 11 by the imaging unit 13. The system supports intracytoplasmic sperm injection (ICSI) based on the imaging video captured by the input tool 3. The following describes the ICSI support device 2. I will explain in detail below.

[0032] (Microinsemination support device) Figure 2 is a functional block diagram of the intracytoplasmic sperm injection support device 2. The intracytoplasmic sperm injection support device 2 includes an acquisition unit. The system comprises a control unit 21, a control unit 22, and an analysis unit 23. The intracytoplasmic sperm injection support device 2 includes a sperm injection tool 3 This supports intracytoplasmic sperm injection (ICSI) by injecting sperm held in an egg into an egg held by an egg retention device. do.

[0033] The acquisition unit 21 captures the oocyte pool PO and sperm injection tool 3 on the stage 11 via the imaging unit 13. By imaging, information including the relative positional relationship between the sperm injection tool 3 and the egg is obtained. The acquisition unit 21, based on the captured image or captured video image captured by the imaging unit 13, determines the sperm injection It is sufficient to acquire information including the relative positional relationship between the input tool 3 and the egg, and the imaging unit 13 will capture It is not necessary to acquire the captured image or the captured video itself.

[0034] In one embodiment, the acquisition unit 21 captures the image captured by the imaging unit 13, and the sperm injection tool is in the imaging field of view. Image or motion image including the 3 and the egg may be acquired. In one embodiment, The acquisition unit 21 obtains sperm injection by analyzing the captured image or captured video image captured by the imaging unit 13. Information showing the relative positional relationship between the sperm injection tool 3 and the egg may be obtained. Information indicating the relative positional relationship with the egg is, for example, the relationship between the entry / exit of the sperm injection tool 3 and the egg. This may be the distance, or the coordinates of the entry / exit point of the sperm injection tool 3 and the egg. Sperm injection tool 3 Information showing the relative positional relationship between the sperm and the egg is, for example, provided by the sperm injection tool 3 and the egg retention device. It is estimated from the relative positional relationship with the markers placed around the part that contains the egg. It is acceptable. The inlet and outlet of the sperm injection tool 3 and the coordinates of the egg are in the xy coordinates in the in-plane direction of stage 11. It may be a target, and furthermore, it may be an xyz coordinate system with the z direction perpendicular to the face of stage 11 added. i. The acquisition unit 21 is located in the direction of extension of the inlet and outlet of the sperm injection tool 3, that is, the sperm injection tool 3 The insertion direction when inserting the device into the egg cell may be obtained.

[0035] The acquisition unit 21 may further acquire information including the position of the polar body of the egg. Therefore, sperm injection into the cytoplasm of an egg is often performed while avoiding the polar body of the egg. The spindle tends to be located near the polar body (first polar body) of the offspring, and upon insertion of sperm injection tool 3... This is to prevent damage to the spindle. Therefore, the acquisition unit 21 acquires the spindle of the egg cell. Further information, including the location, may be obtained.

[0036] Since the position of the polar body of the egg can be obtained from the observation image of the egg, the acquisition unit 21, The imaging unit 13 acquires an image or video of the oocyte, thereby determining the position of the oocyte's polar body. Information including location may be acquired. In one embodiment, the acquisition unit 21 acquires the image captured by the imaging unit 13. By analyzing captured images or captured video images, information indicating the position of the polar body of the egg is obtained. good.

[0037] The acquisition unit 21 contains sperm in the sperm pool PS located in the sperm injection tool 3 and stage 11. Further information, including the relative positional relationship with the sperm, may be obtained.

[0038] In one embodiment, the acquisition unit 21 captures the image captured by the imaging unit 13, and the sperm injection tool is in the imaging field of view. Image or video footage including 3 and sperm may be acquired. In one embodiment, The acquisition unit 21 obtains sperm injection by analyzing the captured image or captured video image captured by the imaging unit 13. Information showing the relative positional relationship between the sperm injection tool 3 and the sperm may be obtained. Information indicating the relative positional relationship with the sperm is, for example, the relationship between the inlet / outlet of the sperm injection tool 3 and the sperm. This could be distance, or the coordinates of the inlet / outlet of the sperm injection tool 3 and the sperm.

[0039] The acquisition unit 21 may be implemented by the communication unit 2d described later.

[0040] Based on the information acquired by the acquisition unit 21, the control unit 22 injects sperm into the egg and displays It controls the movement of stage 11, where the egg retention device is placed for microfertilization. Based on the information acquired by the acquisition unit 21, the control unit 22 controls the immobilized sperm. The movement of stage 11 may be controlled to hold it in the injection tool 3.

[0041] The control unit 22 causes the sperm injection tool 3 to hold the sperm, and injects the held sperm into the egg. To perform microinsemination, the imaging control unit 221, drive control unit 222, and sperm injection tool control unit 22 3. The system may also include a suction device control unit 224 and a water supply device control unit 225.

[0042] The imaging control unit 221 controls the imaging unit 13 and adjusts the imaging area of ​​the imaging unit 13. The imaging area of ​​the imaging unit 221 may be adjusted by adjusting the imaging field of view of the imaging unit 13. When the image unit 13 performs imaging via the microscope system, the imaging control unit 221 controls sperm and / or To image the egg cell with high resolution, the microscope system attached to the imaging unit 13 is configured This may be adjusted. This adjustment may be performed by the person performing the intracytoplasmic sperm injection (ICSI).

[0043] The drive control unit 222 controls the drive unit 12 and moves the stage 11. 22 may move the stage 11 via the electrical mechanism provided in the drive unit 12.

[0044] The sperm injection tool control unit 223 controls the sperm injection tool 3 and performs sperm aspiration and discharge. The sperm injection tool control unit 223 controls the pressure control mechanism of the sperm injection tool 3, and the sperm By adjusting the output of the pump that the injection tool 3 has or that is attached to the sperm injection tool 3 Good. If the sperm injection tool 3 is equipped with means for immobilizing sperm, the sperm injection tool The control unit 223 activates means for immobilizing sperm and in order to immobilize sperm The injection tool 3 may be controlled.

[0045] The suction device control unit 224 and the water supply device control unit 225 are connected to the egg holding device. The suction device and water supply device are controlled separately. Details of the egg retention device will be described later, but the suction The egg is held in the egg-holding device by a pulling device. Additionally, the egg-holding device is supplied with water by a water supply device. The egg held or placed in the chair may be rotated.

[0046] The suction device control unit 224 and the water supply device control unit 225 control the suction device and the water supply device, respectively. The suction device control unit 224 and the water supply device control unit are controlled to perform egg suction and rotation. The oocyte unit 225 aspirates the oocyte based on the information about the oocyte acquired by the oocyte acquisition unit 21, and then... In response, water is supplied.

[0047] The water supply device control unit 225 operates based on the information acquired by the acquisition unit 21, including the position of the polar body of the egg. Then, the egg is rotated so that sperm can be injected using a sperm injection tool from a direction where the polar body is not located. The water supply device may be controlled to enable sperm injection. For this purpose, the water supply device control unit 225 controls the sperm injection device. With the direction in which the 3rd o'clock position is taken as the 3 o'clock direction, the polar body of the oocyte is located approximately at the 12 o'clock or 6 o'clock direction. By controlling the water supply device to deliver water to the egg, the egg can be rotated. The water supply device control unit 225 ensures that the temperature in the container 14 is maintained at a temperature suitable for intracytoplasmic sperm injection. The temperature of the fluid supplied from the water supply device may be controlled. The control of the fluid temperature can be, for example, by controlling the temperature of the water supply. This is done by temperature control means that may be provided in the apparatus. Examples of temperature control means include Examples include heaters and coolers. The temperature control means controls the temperature of the fluid supplied from the water supply device. It may have a function to measure [something].

[0048] The suction device is connected to the egg retention device and is positioned near the suction port of the egg retention device. The device is not particularly limited as long as it can aspirate the egg to be placed. For example, the aspirating device is The pump may be capable of controlling the pressure using the suction device control unit 224.

[0049] Furthermore, the water supply device is connected to the egg holding device, and fluid is supplied from the water outlet of the egg holding device. The device is not particularly limited as long as it is capable of supplying water. The water supply device is, for example, the water supply device control unit 225 The pump may be capable of controlling the flow rate of the fluid being supplied. The water supply device is a pump capable of supplying the fluid The temperature control means may be provided to maintain the temperature within a certain range. Examples include heaters and coolers. The temperature control means is supplied with water from a water supply device. It may have a function to measure the temperature of the fluid. The fluid temperature should be at a temperature suitable for intracytoplasmic sperm injection. It is preferable to maintain the temperature, which is, for example, around 37°C, and 33-40°C. The temperature may be 35-38°C, 36-37°C, or 37°C. The temperature control means is located in the container 14. The temperature of the fluid being supplied may be controlled to maintain a temperature suitable for intracytoplasmic sperm injection (ICSI).

[0050] The control unit 22 is stored in RAM 2b or ROM 2c, which will be described later, and executed by CPU 2a. This may be implemented by a program. Instructions from the control unit 22 are sent via the communication unit 2d to each It is transmitted to the unit and the device.

[0051] The analysis unit 23 analyzes the information acquired by the acquisition unit 21 and assists the control unit 22 in its operation. In this embodiment, the analysis unit 23 is provided in the intracytoplasmic sperm injection support device 2, but intracytoplasmic sperm injection Other parts or devices besides support device 2, such as the imaging unit 13 or the computer connected to the imaging unit 13. It may be equipped with, for example, a sperm recognition unit 231, an egg recognition unit 232, and a sperm The system may include an evaluation unit 233 and an egg evaluation unit 234.

[0052] The sperm recognition unit 231 analyzes the information acquired by the acquisition unit 21 and recognizes sperm. When 1 acquires an image or video, a method for recognizing sperm from the image or video and Therefore, multiple representative images showing typical sperm shapes are prepared, and each frame in the image or video is analyzed. Methods for pattern matching using representative images, or observational images of sperm as training data. One method is to use a trained model that has been trained using this method.

[0053] The egg recognition unit 232 analyzes the information acquired by the acquisition unit 21 and recognizes the egg. When 1 acquires an image or video, a method for recognizing an egg from the image or video and Therefore, multiple representative images showing typical shapes of an egg cell are prepared, and each frame in the image or video is analyzed. Methods for pattern matching using representative images, or observational images of oocytes as training data. One method is to use a trained model that has been trained using this method. Another method is to use an egg retention device. The egg may be recognized based on the marker attached to it.

[0054] The sperm evaluation unit 233 analyzes the information acquired by the acquisition unit 21 and evaluates the quality of each sperm. Sperm quality refers to the fertilization success rate, blastocyst formation rate, implantation rate, and / or when used in intracytoplasmic sperm injection (ICSI). This refers to the properties of each sperm that relate to the success rate of conception, etc. High sperm quality means that intracytoplasmic sperm injection (ICSI) When used in sperm, the fertilization success rate, blastocyst formation rate, implantation rate, and / or birth success rate are relative. This means that it is high. The sperm evaluation unit 233 has at least one indicator related to sperm quality. Sperm quality can be quantified by calculating [a certain value].

[0055] One indicator relating to sperm quality is, but is not limited to, a global indicator. The WHO Laboratory Manual, 6th Edition, published by the Health Organization (WHO) The indicators listed in "The Examination and Processing of Human Semen; Sixth Edition" are cited. Such indicators include, for example, sperm motility, sperm morphology, and sperm At least one degree of DNA fragmentation, or one calculated from two or more of these: It can be listed.

[0056] The sperm evaluation unit 233 calculates at least one indicator related to sperm quality using various methods. For example, sperm motility is the speed of movement and the trajectory of movement over a given time interval. It can be indexed by linearity, etc. The sperm evaluation unit 233 performs at predetermined time intervals. By tracking sperm, the speed and trajectory of their movement are determined based on the sperm's position in each frame. The linearity of the trace can be calculated. Sperm motility is measured using sperm quality and movement as training data. It may also be calculated using a trained model that has been trained in this way.

[0057] Sperm morphology refers to the morphology of sperm captured in each frame of an image or video. It can be indexed based on this. The sperm evaluation unit 233 evaluates multiple sperm for each sperm. Sperm morphology is indexed based on the frame, and these values ​​are averaged to calculate a single index. It is acceptable to release it. Sperm morphology refers to, for example, the shape of the sperm head, the shape of the sperm tail, and any defects thereof. And it can be indexed based on the balance of size, etc. Sperm motility is the quality of sperm It may also be calculated using a learning model that has been trained using quality and form as training data.

[0058] The degree of sperm DNA fragmentation depends on the speed of motility and trajectory over a given time interval. Sperm motility, such as the linearity of the trail, and each frame of the captured image or captured video The morphology of sperm can be comprehensively considered and indexed. The degree of sperm DNA fragmentation The degree of sperm DNA fragmentation, along with sperm motility and / or morphology, is used as training data. It may also be calculated using a trained model that has been trained using the following. The extent of development is measured using stained images obtained by selectively staining sperm that have undergone DNA fragmentation. It is possible.

[0059] The sperm evaluation unit 233 comprehensively evaluates sperm quality by considering two or more of the above indicators. The valued index may be calculated. The sperm evaluation unit 233 evaluates sperm motility, sperm morphology, and After arbitrarily weighting each index related to the degree of sperm DNA fragmentation, You may calculate the average indicator. In doing so, if any of the indicators fall below the threshold, the calculation will be suspended. It is permissible to impose penalties that reduce a certain number from the indicator.

[0060] The egg evaluation unit 234 analyzes the information acquired by the acquisition unit 21 and evaluates the quality of each egg. Egg quality refers to the fertilization success rate, blastocyst formation rate, implantation rate, and / or when used in intracytoplasmic sperm injection (ICSI). This refers to the properties of each egg related to the success rate of conception, etc. High egg quality means that intracytoplasmic sperm injection (ICSI) When used in sperm, the fertilization success rate, blastocyst formation rate, implantation rate, and / or birth success rate are relative. This means that it is high. The egg evaluation unit 234 has at least one indicator related to egg quality. Egg quality can be quantified by calculating [a certain value].

[0061] The oocyte evaluation unit 234, for example, assesses the position, shape, and size of the polar body, spindle, and smooth endoplasmic reticulum. The quality of the oocyte may be evaluated based on the following: the degree of turbidity of the cytoplasm, etc. Oocyte evaluation unit 2 34 uses a learning model trained with egg quality and morphology as training data to study eggs You may evaluate the quality of the offspring.

[0062] The analysis unit 23 is stored in RAM 2b or ROM 2c, described later, and executed by CPU 2a. This may be implemented by a program. The analysis results from the analysis unit 23 are used by the control unit 22. It may be transmitted to each part and device via the communication unit 2d and displayed on the display unit 2f. That's fine.

[0063] Next, the physical configuration of the intracytoplasmic sperm injection support device 2 will be described with reference to Figure 3. This figure shows an example of the physical configuration of the intracytoplasmic sperm injection support device 2. The intracytoplasmic sperm injection support device 2 is a process The CPU (Central Processing Unit) 2a, which corresponds to the sassin, and RAM (Random Access Memory) 2b and ROM, which correspond to the memory portion. (Read-only Memory) 2c, communication unit 2d, input unit 2e, and display unit 2 It has f and . Each of these configurations is connected to each other via a bus so that data can be sent and received from each other. In this example, the functions of the intracytoplasmic sperm injection support device 2 are configured as a single computer. To explain further, the function of the ICSI support device 2 is achieved by combining multiple computers. It may be shown. Also, the configuration shown in Figure 3 is just one example, and the intracytoplasmic sperm injection support device 2 may be other than those shown. It may have these components, or it may not have some of these components.

[0064] CPU2a controls the execution of programs stored in RAM2b or ROM2c. It performs calculations and processing of data. The CPU 2a controls the computer, the acquisition unit 21, the control unit 22 , and is an arithmetic unit that executes a program to function as the analysis unit 23. CPU2a is The input unit 2e and communication unit 2d receive various data, and the data calculation results are displayed on the display unit 2f. It can be displayed or stored in RAM2b or ROM2c.

[0065] RAM2b is a memory unit that allows data to be rewritten, such as semiconductor memory. It may be composed of elements. RAM2b may store the program executed by CPU2a. Note that these are examples, and RAM2b may store other data as well. That's good too.

[0066] ROM2c is a memory unit from which data can be read, such as semiconductor memory. It may be composed of elements. ROM2c, for example, may store data that is not rewritten. stomach.

[0067] The communication unit 2d connects the intracytoplasmic sperm injection support device 2 to other units and devices, such as the sperm injection tool 3, and the drive unit. This is an interface that connects to the moving unit 12, the imaging unit 13, the suction device, and the water supply device. The signal unit 2d may be connected to a network by wire or wireless connection.

[0068] The input unit 2e accepts data input from the user, for example, a keyboard. And may include a touch panel.

[0069] The display unit 2f visually displays the calculation results from the CPU 2a, for example, L It may consist of a CD (Liquid Crystal Display). 2f is information acquired by the acquisition unit 21, for example, an image or video acquired by the imaging unit 13. You may display the image.

[0070] Furthermore, the intracytoplasmic sperm injection support program according to this embodiment includes RAM2b, ROM2c, and other components. It may be stored and provided on a memory medium readable by the computer, and communication unit 2d It may be provided via a communication network connected by the ICSI support device 2. As explained with reference to Figure 2, when CPU2a executes the program according to this embodiment, The functions described above are realized. Note that these physical configurations are examples and do not necessarily represent independent configurations. It does not have to be a complete system. For example, the intracytoplasmic sperm injection support device 2 has a CPU 2a and RAM 2b and / or LSI (Large-Scale Integration) with ROM2c It may also be equipped with.

[0071] (Egg retention device) Next, referring to Figure 4, the egg retention device for use in the intracytoplasmic sperm injection support system 1 The following describes the egg retention device according to this embodiment. This is used to efficiently implement control by the support device 2. Egg retention according to this embodiment. The device may be made of, for example, resin.

[0072] Figure 4 shows a schematic perspective view (Figure 4(A)) and an enlarged view of the egg retention device D1 according to this embodiment. This is a plan view (Figure 4(B)). As shown in Figures 4(A) and (B), the egg retention device D 1 is a recess 151a, 151b, 151 for accommodating egg cells O1, O2, and O3, respectively. c holds the egg cells O1, O2, and O3 in recesses 151a, 151b, and 151c, respectively. It is equipped with suction ports 154a, 154b, and 154c for this purpose. Figure 4(B) shows the recess 15 This is an enlarged plan view of the oocyte O1 held in 1a. The oocyte holding device D1 holds the oocyte O Water outlet 1 for rotating 1, O2, O3 in recesses 151a, 151b, 151c It also includes 53a, 153b, and 153c.

[0073] In Figure 4(A), the three oocytes O1, O2, and O3 each have three recesses 151a, Although contained in 151b and 151c, the oocyte retention device D1 contains one or more oocytes. And it is sufficient if it has a structure to hold it. Below, as shown in Figure 4(A), three eggs When O1, O2, and O3 are each housed in three recesses 151a, 151b, and 151c. This will be explained, but the number of eggs housed and held in the egg retention device D1 is not particularly limited. It will not be done.

[0074] Furthermore, in the following, if it is not necessary to distinguish between the three eggs O1, O2, and O3, then these will be used. They are collectively referred to as oocyte O. The same applies to the other components, and in Figure 4(A), the oocyte retainer Each component of the holding device D1 is corresponding to one of three eggs O1, O2, and O3. The symbols a, b, and c are assigned, but each corresponds to the three egg cells O1, O2, and O3. If it is not necessary to distinguish between components, the signs a, b, and c are omitted. For example, three eggs. The three recesses 151a, 151b, and 151c, corresponding to O1, O2, and O3, are collectively referred to as recesses. There are cases where it is referred to as section 151.

[0075] Furthermore, the egg retention device D1 according to this embodiment is provided as the egg pool PO in Figure 1. It may be located adjacent to the oocyte pool PO. That is, the oocyte holding device. Each recess in D1 may also be used as an egg pool PO, and the eggs may be stored separately from the recesses of the egg retention device D1. An egg pool PO containing the egg may be provided. Therefore, egg retention according to this embodiment Device D1 may move in accordance with stage 11. Sperm injection tool 3 is located on imaging unit 13. If it is fixedly installed, the imaging unit 13 can be moved by moving the stage 11. In the field of view, the sperm injection tool 3 is almost fixed, and only the egg retention device D1 is moved. This allows for highly accurate intracytoplasmic sperm injection (ICSI) by moving stage 11. can.

[0076] Returning to Figure 4, the oocyte O is housed in the recess 151 of the oocyte holding device D1. The part 151 is not particularly limited as long as it can accommodate the egg O in a predetermined region. For example, it has a width and height of 100 to 1000 μm, preferably 150 to 500 μm. It may be. The recess 151 is, for example, 100 to 1000 μm, preferably 150 to 500 μm. It may have a depth of m.

[0077] The recess 151 may be a recess enclosed on the bottom and all four sides, with an opening at the top. The four-sided enclosure forming 51 may have one side partially removed. By partially removing one side of the four-sided enclosure that forms 151, the egg O and sperm injection can be performed. This makes it easier to bring the 3rd wheel into contact with the other wheel.

[0078] In one embodiment, the recess 151 is surrounded by a bottom surface and a wall extending upward from the bottom surface. It may be formed by such means, and at least a part of the wall portion may be lower in height than other parts of the wall portion. It may be configured in such a way. For example, a part of the wall may be, for example, 100 to 1000 μm thick. Preferably, it may have a height of 150 to 500 μm, and the portion of the wall is relatively low in height. The wall may have a height of, for example, 20 to 200 μm, preferably 50 to 150 μm. The portion with a relatively low height may be located on the opposite side of the wall where the suction port, described later, is provided. .

[0079] A suction port 154 is provided in the wall portion that forms the recess 151 of the egg-holding device D1. The egg cell O is then collected by the suction port 154 and held in the egg cell holding device D1. This is possible. With the egg O aspirated by the suction port 154, the sperm injection tool 3 is inserted. By bringing the outlet into contact with the egg O, sperm can be injected into the egg O. The drawpipette 154 serves the role of a holding pipette in conventional intracytoplasmic sperm injection procedures.

[0080] Therefore, the suction port 154 has a shape similar to that of the suction port of a holding pipette. It is acceptable. For example, as shown in Figure 4(B), the suction port 154 is positioned closer to the recess 151. Therefore, it may have a tapered shape that reduces in diameter. The shape of the opening of the suction port 154 is particularly The shape may be, for example, approximately circular, approximately elliptical, or a rounded rectangle. By doing so, the damage to the egg O during suction can be reduced. The circular diameter d of the opening of the suction port 154 is, for example, 10 to 150 μm, preferably 15 to 1 It is 00 μm. The equivalent circle diameter refers to the diameter of a circle that has the same area as the shape of the aperture.

[0081] The suction port 154 is connected to a suction device for aspirating the egg O through the suction port 158. The suction opening 154 and the suction port 158 ​​are connected by a flow path inside the egg holding device D1. It is continued. The suction port 158 ​​corresponds to multiple suction ports 154a, 154b, 154c. Multiple suction ports may be provided, and each of the multiple suction ports 154a, 154b, and 154c may be connected to one another. One or more suction ports 158 may be provided. In Figure 4(A), multiple suction ports are provided. Each of the openings 154a, 154b, and 154c is connected to a single suction port 158. Therefore, in Figure 4(A), the suction device connected to the suction port 158 ​​is activated. When egg cells O are aspirated, the egg cells O contained in recesses 151a, 151b, and 151c O1, O2, and O3 will all be aspirated simultaneously.

[0082] Furthermore, as shown in Figure 4(B), when aspirating the oocyte O through the suction port 158, A fluid flow F1 is generated from the opening of the intake port 154 toward the suction port 158, and the oocyte O is suctioned into the suction port It is attracted to the 154 opening and comes into contact with the opening. By maintaining suction in this state, the egg O can be held in the opening of the suction port 154.

[0083] A water inlet 153 is provided in the wall portion that forms the recess 151 of the egg-holding device D1. It is being done. By supplying fluid from the water inlet 153, the eggs contained in the recess 151 are pushed into the recess. The part 151 can be moved or rotated.

[0084] As mentioned above, in intracytoplasmic sperm injection (ICSI), a sperm injection tool is used to avoid spindle damage. When inserting into the egg, it is preferable to avoid the polar body. As shown in Figure 4(B), the egg O The polar body PB position information acquisition unit 21 acquires the position information while water is supplied to the egg O from the water inlet 153. By doing so, the polar body is moved to the appropriate position and held in this state at the opening of the suction port 154. It is possible.

[0085] In the embodiment shown in Figure 4(B), the sperm injection tool 3 is directed in the opposite direction to the suction port 154, that is, Since it is inserted from below in Figure 4(B), the polar body PB is directed towards the water inlet 153 or the drain outlet 155. In other words, it is preferable that it be located to the left or right of Figure 4(B).

[0086] The shape of the water inlet 153 is not particularly limited as long as it can supply fluid to the recess 151. Furthermore, the fluid supplied from the water inlet 153 is typically a liquid such as water or culture medium, but air A gas such as air may be supplied to the recess 151.

[0087] The water inlet 153 is a water supply device for supplying water to the egg O through the water supply port 157. It is connected to the water inlet 153 and the water supply port 157, which form a flow path inside the egg holding device D1. It is connected by multiple water outlets 153a, 153b, 153c Multiple outlets may be provided to correspond to each of the multiple water outlets 153a, 153b, 153c One or more water supply ports 157 connected to them may be provided. In Figure 4(A), multiple One water supply port 15 connected to each of the number of water inlets 153a, 153b, and 153c 7 is provided. Therefore, in Figure 4(A), the water supply device connected to the water supply port 157 When the device is activated and egg cells O are aspirated, the eggs are sent to recesses 151a, 151b, and 151c. Water will be used.

[0088] Furthermore, as shown in Figure 4(B), when water is supplied to the recess 151 through the water supply port 157... A fluid flow F2 is generated from the water supply port 157 toward the opening of the water supply inlet 153.

[0089] Even if a drainage port 155 is provided in the wall portion forming the recess 151 of the egg-holding device D1 Good. The drain port 155 is provided as a discharge path for the fluid supplied from the water inlet 153. If the upper part of the recess 151 is made into an opening as shown in Figure 4(A), the drain 155 should be omitted. It is possible.

[0090] The drain outlet 155 may be connected to a drainage device through the drain port 159. The drainage port 159 is connected by a flow path inside the egg retention device D1. Even if multiple ports 159 are provided to correspond to multiple drain outlets 155a, 155b, and 155c Often, one or more drains are connected to each of the multiple drains 155a, 155b, and 155c. A drain port 159 may be provided. As shown in Figure 4(B), through the drain port 159 When draining, the fluid flow F3 from the opening of the drain outlet 155 toward the drain port 159 To occur.

[0091] Around the recess 151 of the egg retention device D1, there is a feature that assists in the recognition of the recess 151 and the egg O. A marker 152 may be attached. There may be one or more markers 152. A wall is provided with a suction port 154 to facilitate contact between the egg O and the sperm injection tool 3. It is preferable that a marker 152 is provided on the wall opposite to the part. In this case, the control unit 22 The system recognizes the marker 152 and moves the sperm injection tool 3 from the direction of the marker 152. The marker 152 can be controlled to insert into the egg cell O. It may be provided around the oocyte, in which case the analysis unit 23 recognizes the marker 152 and It is possible to recognize the approximate location of O.

[0092] [Intracytoplasmic sperm injection (ICSI) method] The following describes a method for performing intracytoplasmic sperm injection (ICSI) using the ICSI support system according to this embodiment. The intracytoplasmic sperm injection method according to this embodiment is described by the intracytoplasmic sperm injection support system according to this embodiment. Because of this, at least some of the steps in intracytoplasmic sperm injection can be automated. In particular, the oocyte Because the process of injecting sperm can be automated, the individual skills of the embryologist can be automated. In vitro fertilization can be performed efficiently and with a high success rate.

[0093] Figure 5 is a flowchart showing an example of the steps of the intracytoplasmic sperm injection method according to this embodiment. Oh, although not shown in Figure 5, the person performing the intracytoplasmic sperm injection (typically an embryologist) in this embodiment A container 14 containing a sample is placed on stage 11 of the intracytoplasmic sperm injection support system 1, and imaging is performed. The settings are configured so that the sample and the entry / exit points for the sperm injection tool 3 are located within the imaging field of view of section 13. .

[0094] In the following, the intracytoplasmic sperm injection (ICSI) support system 1 automatically performs the ICSI procedure without requiring any processing from the person performing the ICSI. This section explains how to perform intracytoplasmic sperm injection (IVF), but some or all of the following steps are performed using intracytoplasmic sperm injection (ICSI). This can also be done manually by the person implementing it.

[0095] As shown in Figure 5, in the microinsemination method according to this embodiment, first, the imaging field of the imaging unit 13 The sperm are moved to the sperm pool PS (hereinafter also referred to as "step S1"). Step S1 is, The imaging control unit 221 controls the imaging unit 13, and / or the drive control unit 222 controls the drive unit 12. By maneuvering and moving stage 11, the imaging unit 13 can see the sperm pool PS on stage 11. To image the area.

[0096] In step S1, the location of the sperm pool PS is stored in the intracytoplasmic sperm injection support device 2. This retrieves the position of the sperm pool PS that was stored in memory, and the imaging area of ​​the imaging unit 13 is set to the sperm pool You may move them to the sperm pool PS. Alternatively, you can place a marker indicating the location of the sperm pool PS on the sperm pool By placing the marker around the PS, the analysis unit 23 recognizes the marker, and the imaging unit 1 The imaging area of ​​3 may be moved to the sperm pool PS.

[0097] Next, sperm to be used for intracytoplasmic sperm injection (ICSI) are selected and immobilized (hereinafter also referred to as "Step S2"). (u) In step S2, first, good quality sperm suitable for use in intracytoplasmic sperm injection (ICSI) are selected. Specifically, the sperm recognition unit 231 recognizes each sperm in the sperm pool PS, and the sperm evaluation unit 233 The sperm evaluation unit 233 evaluates the quality of each sperm. Sperm are selected. The sperm evaluation unit 233 selects sperm that are of relatively high quality from among the sperm whose quality has been evaluated. You may select sperm that have been evaluated as such, or you may select sperm that exceed a predetermined threshold. stomach.

[0098] In step S2, the selected sperm are then immobilized. Sperm immobilization is performed before sperm injection. The tool control unit 223 may control and perform the immobilization means provided in the sperm injection tool 3. Alternatively, the control unit 22 may control and perform the procedure using an immobilization device different from the sperm injection tool 3. The sperm injection tool control unit 223 controls the immobilization means provided in the sperm injection tool 3 to control the sperm To immobilize the sperm, the selected sperm and the immobilization means provided in the sperm injection tool 3 are immobilized. In order to achieve the appropriate positional relationship, the drive control unit 222 controls the drive unit 12, and the stage 11 You may move it. If you are immobilizing the sperm by pressing the inlet / outlet of sperm injection tool 3 directly against the sperm, In order to properly bring the inlet and outlet of the sperm injection tool 3 into contact with the sperm, the drive control unit 222 controls the drive unit You may control 12 and move stage 11.

[0099] Next, the sperm selected and immobilized in step S2 are aspirated (hereinafter referred to as "step S3"). (Also known as ") In step S3, bring the sperm to be aspirated close to the inlet and outlet of the sperm injection tool 3. To do this, the drive control unit 222 controls the drive unit 12 and moves the stage 11. The sperm injection tool control unit 223 controls the sperm injection tool 3, and sucks sperm from the inlet and outlet. The sperm is held in the sperm injection tool 3.

[0100] Next, the imaging field of view of the imaging unit 13 is moved to the oocyte pool (hereinafter referred to as "step S4") (Also known as) Step S4 is when the imaging control unit 221 controls the imaging unit 13 and / or drives it. The control unit 222 controls the drive unit 12 to move the stage 11, so the imaging unit 13 moves the stage Image the oocyte pool PO on page 11.

[0101] In step S4, the position of the oocyte pool PO is stored in the intracytoplasmic sperm injection support device 2. This retrieves the stored position of the egg pool PO, and the imaging area of ​​the imaging unit 13 is set to the egg pool You may move them to the oocyte pool PO. Alternatively, you can place a marker indicating the location of the oocyte pool PO in the oocyte pool. By placing the marker around the PO, the analysis unit 23 recognizes the marker, and the imaging unit 1 The imaging area of ​​3 may be moved to the oocyte pool PO. The analysis unit 23 analyzes the oocyte holding device It may recognize the marker attached to the egg retention device, or it may recognize the marker attached to the egg retention device.

[0102] Next, the eggs to be used for intracytoplasmic sperm injection (ICSI) are identified and selected (hereinafter also referred to as "Step S5"). Step S5 involves selecting good quality eggs suitable for use in intracytoplasmic sperm injection (ICSI). Specifically The oocyte recognition unit 232 recognizes each oocyte in the oocyte pool PO, and the oocyte evaluation unit 234 evaluates each oocyte. The quality of the eggs is evaluated. The egg evaluation unit 234 selects good quality eggs from among the eggs whose quality has been evaluated. Select. The egg evaluation unit 234 selects from among the eggs whose quality has been evaluated that are evaluated to be of relatively high quality. You may select eggs that have been fertilized, or eggs that exceed a predetermined threshold.

[0103] In intracytoplasmic sperm injection (ICSI), the number of egg cells prepared is often less than the number of sperm cells. Therefore, to increase the success rate of fertilization, high-quality sperm should be injected into high-quality eggs. It is preferable to do so. In Figure 5, steps S4 and S5 are performed before step S1. You can then adjust the sperm quality selected in step S2 according to the quality of the selected egg. stomach.

[0104] For example, if a good quality egg is selected in step S5, then in step S2, Either select high-quality sperm or set the sperm selection threshold higher than the normal setting. You can set up a system to select high-quality sperm.

[0105] Alternatively, in intracytoplasmic sperm injection (ICSI), sperm is often injected into all of the prepared eggs. Therefore, the evaluation of egg quality can be omitted.

[0106] Alternatively, if inferior quality eggs are selected in step S5, for example, the quality will be shown If an egg is selected whose indicators fall below the threshold, the automated sperm injection process will be discontinued, and intracytoplasmic sperm injection (ICSI) will be performed. You may also notify the implementer and encourage them to perform the injection manually.

[0107] Next, the orientation of the egg selected in step S5 is adjusted (hereinafter also referred to as "step S6"). (Says). In step S6, first, the position of the sperm injection tool 3 and the egg acquired by the acquisition unit 21 is determined. Based on the positional relationship, the analysis unit 23 determines the preferred orientation of the egg. For example, the analysis unit 23 The acquisition unit 21 recognizes the position of the polar body of the acquired egg, and determines the direction in which the sperm injection tool 3 will be inserted. The orientation of the egg is determined so that its polar body is positioned perpendicular to the target.

[0108] Next, the egg oriented in the direction determined in step S5 is held in the egg holding device ( (Also known as "Step S7"). In Step S7, the water supply device control unit 225 controls the water supply device The suction device control unit 224 controls the suction device while controlling the direction, thereby directing it in the appropriate direction. The eggs are then held in an egg retention device.

[0109] Next, the sperm aspirated in step S3 is injected into the egg held in step S7 (see below) (Also known as "Step S8"). In Step S8, sperm is first injected into the egg for intracytoplasmic sperm injection (ICSI). To perform sperm injection, the egg is brought close to the inlet and outlet of the sperm injection tool 3. At this time, the drive control unit 2 Unit 22 controls the drive unit 12 and moves the stage 11. Next, the sperm injection tool control unit. 223 controls the sperm injection tool 3 and injects sperm into the egg through the inlet and outlet. Injection is complete. Next, the sperm injection tool 3 is removed from the egg, completing the intracytoplasmic sperm injection (ICSI) procedure.

[0110] Next, it is determined whether sperm injection has been completed into all eggs (hereinafter referred to as "Step S"). Step S9 is also called "9". Step S9 should be performed when the number of prepared egg cells is small. If a very large number of eggs are available, this step can be omitted. The intracytoplasmic sperm injection support device 2 is By remembering which eggs have been injected with sperm, it is possible to detect the presence of eggs that have not been injected with sperm. If the procedure is refused (Step S9: No), return to Step S1. At this time, sperm is injected. The number of missing eggs may be displayed on display unit 2f. Sperm injection is complete for all eggs. If it is determined that the process is complete (Step S9: Yes), the process is terminated.

[0111] In addition, in each of the above steps, the acquisition unit 21 optionally performs the following: Information including the relative positional relationship between the entry / exit point and the egg, and / or the entry / exit point of sperm injection tool 3 Information including the relative positional relationship with the sperm may be obtained. For example, in step S3, Information including the relative positional relationship between the sperm injection tool 3 and the sperm is obtained, and based on this, Sperm are aspirated using sperm injection tool 3. Also, in step S8, sperm injection tool 3 Information including the relative positional relationship between the entry / exit point and the egg is obtained, and based on this, sperm injection is performed. Insert the 3 into the egg. Also, in step S6, the acquisition unit 21 is the polar body or Further information, including the position of the spindle, is acquired. In addition, the acquisition unit 21 acquires the image captured from the imaging unit 13. When acquiring an image or video, in each step, the sperm injection tool 3 is placed in the imaging field. If at least one of an egg and sperm is included, based on the captured image or captured video The coordinates of at least one of the sperm injection tool 3, the egg, and the sperm may be obtained.

[0112] [Differentiation] Although an example of this embodiment has been described above, the present invention is not limited to the above. i. For example, examples and / or preferred embodiments in the description of each component and step (preferred (More desirable, more preferable, even more preferable, even more preferable, etc.) are not specifically mentioned. Unless otherwise specified, each can be arbitrarily combined to form this embodiment. For example, each Regarding the implementation form, the embodiment described as a preferred embodiment and the embodiment described as a preferred embodiment The embodiments described may be combined with other embodiments, or are described as preferred embodiments, etc. The embodiments and more preferred embodiments (or even more preferred embodiments, etc.) are described below. The configuration may be combined with the existing configuration. Furthermore, each component and step is effective in this embodiment. It can be omitted as appropriate, as long as it does not hinder the fruit.

[0113] For example, in the intracytoplasmic sperm injection support system 1 shown in Figure 1, the egg pool PO and Although a sperm pool PS is provided, the egg pool PO and sperm pool PS are in different containers. It may be provided in [location]. Also, the intracytoplasmic sperm injection support system 1 may be used only for injecting sperm into the egg. In this case, a sperm pool does not need to be provided in stage 11. In this case, intracytoplasmic sperm injection (ICSI) The user of support system 1 (typically an embryologist) takes the sperm infusion tool 3 containing the sperm. In this case, perform intracytoplasmic sperm injection (ICSI). In this case, follow step S1 of the flowchart shown in Figure 5. ~S3 can be omitted.

[0114] Furthermore, in the above description, the sperm injection tool 3 is fixed to the imaging unit 13, but sperm This does not prevent the addition of a mechanism to move the injection tool 3. The intracytoplasmic sperm injection support device 2, relating to the method of administration, has a sperm injection tool control unit 223 that controls the sperm injection tool You may control the movement of Rule 3. In this case, the movement of Stage 11 may be the primary means of movement (for example) It can be used as a means for aligning the egg and the entry / exit port of the sperm injection tool 3 in the translational direction. , the movement of the sperm injection tool 3 is aided by a means of movement (for example, the entry and exit points of the sperm injection tool 3 are connected to the egg) It may be used as a rotational alignment means for adjusting the direction in which it is inserted.

[0115] Furthermore, in the intracytoplasmic sperm injection method according to this embodiment, intracytoplasmic sperm injection may be performed before, during, or after each step. The processing of sperm by the person performing the procedure may be accepted. For example, in step S2, the procedure of intracytoplasmic sperm injection. The person performing the in vitro fertilization may select the sperm to be aspirated, or the person performing the in vitro fertilization may immobilize the sperm.

[0116] Furthermore, in step S3, the person performing the intracytoplasmic sperm injection may specify the method of sperm aspiration. For example, depending on whether the person performing the in vitro fertilization procedure aspirates the sperm from the head end or the tail end... This option may be selected.

[0117] Furthermore, in step S5, the person performing the intracytoplasmic sperm injection may select the egg into which the sperm will be injected. .

[0118] Furthermore, in the injection of sperm into the egg in steps S6-S8, the person performing the intracytoplasmic sperm injection (ICSI) Adjustments may be made. For example, the part of the egg into which the sperm injection tool 3 is inserted. The person performing the in vitro fertilization procedure will determine which direction to insert the sperm from and how far to insert it. You may specify this.

[0119] Furthermore, in the preceding or during each step, the sperm and / or eggs are re-analyzed. To capture images at a high resolution, the imaging control unit 221 is attached to the imaging unit 13 and / or the imaging unit 13. The settings of the microscope system may be adjusted. This adjustment is performed by the person performing the intracytoplasmic sperm injection. This is also good. For example, depending on the object and purpose of imaging at each step, the observation method and magnification in the optical microscope may vary. The rates may be adjusted to ensure that each step is carried out appropriately.

[0120] Furthermore, before each step, the person performing the in vitro fertilization procedure is asked whether they wish to proceed to the next step. It may include a confirmation step. Also, when an abnormality is detected in each step... It is also possible to switch to processing by the person performing the intracytoplasmic sperm injection. For example, if the egg is in an unusual state, such as if the perivitelline space is filled and the cytoplasm is not properly formed. Examples include cases where it cannot be recognized, or where there is an abnormality in the shape of the zona pellucida. At that time, the person performing the intracytoplasmic sperm injection (ICSI) will be instructed to confirm the setup of the ICSI support system 1. Knowledge may be displayed.

[0121] In addition to the embodiment shown in FIG. 4, the egg holding device may also be the embodiment shown in FIG. 6. The egg holding device D2 shown in FIG. 6 is different from the egg holding device D1 shown in FIG. 4, and the roles of the first recess 251a, the second recess 251b, and the third recess 251c are different from each other. <​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​

[0126] The egg retention device D2 further contains the egg O1 housed in the first recess 251a in the second recess It is equipped with a first water inlet 253a for transfer to 251b. The egg retention device D2 has the first By supplying fluid from the water inlet 253a, the oocyte O contained in the first recess 251a 1 can be transferred to the second recess 251b. This allows sperm to be injected into the egg O2. After completion, sperm can be injected into egg cell O1.

[0127] In the egg retention device D2, the second water inlet 253b delivers the egg O2 to the second recess 251b In addition to its role in rotation, after sperm injection into the egg O2 is complete It serves to transfer the fluid supplied from the second water inlet 253b to the third recess 251c. By adjusting the amount and speed, the oocyte O2 is rotated in the second recess 251b, This allows us to distinguish between cases where the oocyte O2 is transferred to the third recess 251c.

[0128] Furthermore, the first water inlet 253a and the second water inlet 253b are similar to those of the egg retention device D1. It is connected to the water supply device via the water supply port 257. Similar to the egg retention device D1, The first water outlet 253a and the second water outlet 253b are conveniently connected to the same water supply port 257. They may be connected to different water supply ports 257.

[0129] Furthermore, the egg retention device D2 includes a marker 252, a suction port 254, a drain port 255, and a suction port. It is equipped with port 258 and drainage port 259, but its configuration is that of the egg retention device D1 Marker 152, suction port 154, drain port 155, suction port 158, and drain port 15 It may be the same as 9.

[0130] [Supplementary Note] The present disclosure includes the following embodiments. [1] A micromanipulation support device for assisting micromanipulation in which sperm held by a sperm injection tool is injected into an egg held by an egg holding device, comprising: an acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg; a control unit that controls the movement of a stage on which the egg holding device is disposed to inject the sperm into the egg based on the information acquired by the acquisition unit and perform micromanipulation; and , [[ID=--]] A micromanipulation support device. [2] The acquisition unit further acquires information including the relative positional relationship between the sperm injection tool and sperm contained in a sperm pool disposed on the stage, and the control unit further controls the movement of the stage to hold the immobilized sperm in the sperm injection tool based on the information acquired by the acquisition unit. The micromanipulation support device according to [1]. [1] [3] The micromanipulation support device according to [1] or [2], further comprising: [[ID=--]] an imaging unit that images the egg, the sperm, and the sperm injection tool; and a driving unit that moves the stage based on an instruction from the control unit. [1] or [2]. [4] The egg holding device is connected to a suction device for holding the egg in the egg holding device, <-- and the control unit further controls the suction device. The micromanipulation support device according to any one of [1] to [3]. [5] The egg holding device is for rotating the egg in the egg holding device, Connected to the water supply device, The control unit further controls the water supply device. An intracytoplasmic sperm injection support device described in any one of [1] to [4]. [6] The acquisition unit further acquires information including the position of the polar body of the egg, The control unit injects the sperm using the sperm injection tool from a direction in which the polar body is not located. To rotate the egg in order to enter, the water supply device is controlled. [5] The intracytoplasmic sperm injection support device described below. [7] The water supply device includes temperature control means for maintaining the temperature of the water being supplied within a certain range. El, The intracytoplasmic sperm injection support device described in [5] or [6]. [8] The sperm injection tool is fixedly mounted to the imaging unit. An intracytoplasmic sperm injection support device as described in any one of [3] to [7]. [9] Egg retention data for placement in an intracytoplasmic sperm injection support device as described in any one of [1] to [8] It is a vice, A recess for containing the egg, A suction port for holding the egg in the recess, Equipped with, Egg retention device.

[10] The device further includes a water inlet for rotating the egg in the recess. The egg retention device described in [9].

[11] A microinsemination support device described in any one of [1] to [8], The aforementioned sperm injection tool, Equipped with, Intracytoplasmic sperm injection (ICSI) support system.

[12] Sperm held in a sperm injection tool are injected into an egg held by an egg retention device. This is an intracytoplasmic sperm injection (ICSI) support program that assists with ICSI, Computers, An acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg, Based on the information acquired by the acquisition unit, the sperm is injected into the egg and microscopically implanted. A control unit that controls the movement of the stage on which the egg-holding device is placed for fertilization, and , To make it function as program. [Explanation of Symbols]

[0131] 1... ICSI support system, 2... ICSI support device, 3... Sperm injection tool, 11... Ste Page, 12...Drive unit, 13...Imaging unit, 14...Container, 21...Acquisition unit, 22...Control unit, 23... Analysis section, 151, 251... recessed area, 152, 252... marker, 153, 253... water inlet, 154,254...Suction port, 155,255...Drain port, 157,257...Water supply port, 15 8,258... Suction port, 159,259... Drainage port, 221... Imaging control unit, 222... Drive control unit, 223... Sperm injection tool control unit, 224... Suction device control unit, 225... Water supply unit Control unit, 231...Sperm recognition unit, 232...Egg recognition unit, 233...Sperm evaluation unit, 234...Egg Child evaluation unit, D1, D2... Oocyte holding device, O... Oocyte, PO... Oocyte pool, PS... Sperm pool Ru.

Claims

1. Sperm held in a sperm injection tool are injected into an egg held by an egg retention device. An intracytoplasmic sperm injection (ICSI) support device that assists in intracytoplasmic sperm injection, An acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg, Based on the information acquired by the acquisition unit, the sperm is injected into the egg and microscopically implanted. A control unit that controls the movement of the stage on which the egg-holding device is placed for fertilization, and 、 An intracytoplasmic sperm injection support device equipped with the following features.

2. The acquisition unit is contained in the sperm pool arranged in the sperm injection tool and the stage. Further information, including the relative positional relationship with the sperm, is obtained. The control unit further determines the immobilized pre Controlling the movement of the stage in order to retain the recorded sperm in the sperm injection tool. The intracytoplasmic sperm injection support device according to claim 1.

3. An imaging unit for imaging the egg, the sperm, and the sperm injection tool, A drive unit for moving the stage based on instructions from the control unit, Furthermore, The intracytoplasmic sperm injection support device according to claim 1.

4. The egg holding device is an aspiration device for holding the egg in the egg holding device. Connected to The control unit further controls the suction device. The intracytoplasmic sperm injection support device according to claim 1.

5. The egg holding device is for rotating the egg in the egg holding device. Connected to the water supply device, The control unit further controls the water supply device. The intracytoplasmic sperm injection support device according to claim 1.

6. The acquisition unit further acquires information including the position of the polar body of the egg, The control unit injects the sperm using the sperm injection tool from a direction in which the polar body is not located. To rotate the egg in order to enter, the water supply device is controlled. The intracytoplasmic sperm injection support device according to claim 5.

7. The water supply device includes temperature control means for maintaining the temperature of the water being supplied within a certain range. El, The intracytoplasmic sperm injection support device according to claim 5.

8. The sperm injection tool is fixedly mounted to the imaging unit. The intracytoplasmic sperm injection support device according to claim 3.

9. Egg retention device for placement in the intracytoplasmic sperm injection support device according to any one of claims 1 to 8 It is a chair, A recess for containing the egg, A suction port for holding the egg in the recess, Equipped with, Egg retention device.

10. The device further includes a water inlet for rotating the egg in the recess. The egg retention device according to claim 9.

11. An intracytoplasmic sperm injection support device according to any one of claims 1 to 8, The aforementioned sperm injection tool, Equipped with, Intracytoplasmic sperm injection (ICSI) support system.

12. Sperm held in a sperm injection tool are injected into an egg held by an egg retention device. This is an intracytoplasmic sperm injection (ICSI) support program that assists with ICSI, Computers, An acquisition unit that acquires information including the relative positional relationship between the sperm injection tool and the egg, Based on the information acquired by the acquisition unit, the sperm is injected into the egg and microscopically implanted. A control unit that controls the movement of the stage on which the egg-holding device is placed for fertilization, and 、 To make it function as program.

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