JP6882713B2 - Cell quality evaluation system, program and cell quality evaluation method - Google Patents

Description

The present invention relates to, for example, a cell quality evaluation system, a cell quality evaluation program, and a cell quality evaluation method used for evaluating the superiority or inferiority of the quality of cultured cells.

In recent years, in the medical field such as regenerative medicine and infertility treatment, it is necessary to prevent the use of cells other than those satisfying predetermined quality requirements in order to realize the desired effect of cell therapy while ensuring safety. .. For example, when a fertilized egg is prepared, cultured and transplanted for in vitro fertilization, the state of the fertilized egg during cultivation is observed at regular intervals to evaluate whether or not the fertilized egg is developing smoothly. At the same time, fertilized eggs that do not meet the prescribed developmental conditions and quality are excluded from the transplant target, or the priority is lowered, and specialists such as doctors take into account the patient's condition in addition to the quality of the fertilized egg. It is necessary to make a final decision, such as transplanting a fertilized egg in good condition, in order to ensure safety.

Therefore, in the conventional medical field, an operator such as a doctor captures an image for observing cells using a microscope, and while checking the image with the naked eye, the cells are based on the experience and knowledge of each operator. The superiority and inferiority of the quality of the cells are evaluated, and cells suitable for transplantation are selected.

Further, in recent years, in order to reduce the burden on the operator and homogenize the determination of cell quality, images showing the state of chromosomes in cells are taken at predetermined time intervals, and based on the images, evaluation targets are used. A state stage identification method (for example, Patent Document 1) that automatically identifies a stage of the internal state of a cell (hereinafter, referred to as “state stage”) that transitions at predetermined time intervals, and an image is taken. A cell quality evaluation system (for example, Patent Document 2) has been proposed in which a feature amount in a cell image is extracted and the superiority or inferiority of the cell quality is determined from the extracted feature amount or a combination of a plurality of feature amounts. ..

International Publication No. WO2006 / 061958 Patent No. 4852890

However, in the method described in Patent Document 1, although the state stage of the cell can be automatically specified, it is difficult to appropriately evaluate the quality of the cell.

Further, in the system described in Patent Document 2, it is not possible to evaluate the cell quality reflecting the internal state of the cell at each state stage at the time of the cell quality evaluation, and it is difficult to improve the accuracy of the quality evaluation.

The present invention has been made in view of the above circumstances, and an object of the present invention is to evaluate the quality of cells to be evaluated automatically and with high accuracy by quantifying the quality of the cells to be evaluated. It is to provide a cell quality evaluation system and the like capable of this.

In order to solve the above-mentioned problems, in the cell quality evaluation system of the present invention, the state stage indicating the internal state gradually changes according to the passage of time from the fertilization of the sperm cell to the egg cell , and the transition of the state stage changes. (1) At least the first state stage in which the progenitor membrane, the transparent band, and the polar body are generated, (2) the second state stage in which the pronucleus is generated in the progenitor membrane, (3) ) A third state stage in which the pronucleus disappears and egg splitting occurs sequentially inside the transparent band, (4) the transparent band and the progenitor membrane disappear, and vegetative membrane cells and internal cell clusters develop. In a cell quality evaluation system that evaluates the quality of cells that transition in the order of the fourth state stage that has changed to a cyst, by controlling the imaging means that images the cells and generates image data. Corresponds to the imaging control means that captures an image of the cell at a predetermined timing from fertilization and generates the image data corresponding to each of the first to fourth state stages, and the generated image data. serving as a reference the scoring criterion information when evaluating the score of the quality of cells in each state stage based on the image of the cell is, in correspondence with the status stage identification information for identifying the respective status stage, it is recorded and recording control means for controlling the recording means, and obtaining means for obtaining image data output from the imaging means, for each of the state stage, while a predetermined image processing on the acquired cell image, A scoring means for scoring the quality of the corresponding cell based on the relevant scoring criterion information, and an evaluation means for evaluating the superiority or inferiority of the quality of the cell according to the scoring result indicating the scored result. It has a configuration including a presentation means for presenting the evaluation result in the evaluation means to the user so as to be viewable.

With this configuration, the cell quality evaluation system of the present invention can automatically evaluate the quality of the target cell by imaging the cell over time, so that the user at the time of cell quality evaluation can be evaluated. Since the workload of (for example, a doctor or an operator) can be reduced, and the superiority or inferiority of cell quality can be evaluated by scoring cells that are in different states for each state stage in each state stage. , It is possible to accurately evaluate cells whose growth process is important in quality evaluation.

That is, since the internal structure of the cell changes according to the state stage and the quality criterion also changes according to the state stage, the cell quality evaluation system of the present invention has different scoring standard information and algorithms for each state stage. It is possible to perform scoring while applying the above, and it is possible to realize appropriate scoring according to the state stage.

Therefore, the cell quality evaluation system of the present invention can quantify the quality of the cell to be evaluated, and can automatically and highly accurately evaluate the cell quality.

The cell quality evaluation system of the present invention can quantify the quality of cells to be evaluated and can evaluate the quality automatically and with high accuracy.

It is a system block diagram which shows the structure of one Embodiment of the cell quality evaluation system which concerns on this invention. It is a figure for demonstrating the transition of the state stage which occurs in a fertilized egg with the lapse of time from insemination when culturing a fertilized egg in the cell quality evaluation system of one embodiment. It is a block diagram which shows the structure of the server apparatus of one Embodiment. It is a figure which shows an example of the data recorded in the cultured cell management DB provided in the server apparatus of one Embodiment. It is a figure which shows an example of the data recorded in the state stage management DB provided in the server apparatus of one Embodiment. It is a figure which shows an example of the data recorded in the scoring information DB provided in the server apparatus of one Embodiment. It is a figure (the 1) explaining the outline of the scoring process in the state stage (first stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (No. 2) explaining the outline of the scoring process in the state stage (first stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (the 1) explaining the outline of the scoring process in the state stage (second stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (No. 2) explaining the outline of the scoring process in the state stage (second stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (the 1) explaining the outline of the scoring process in the state stage (third stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (No. 2) explaining the outline of the scoring process in the state stage (third stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (the 3) explaining the outline of the scoring process in the state stage (third stage) of a fertilized egg in the cell quality evaluation system of one embodiment. It is a figure (the 1) explaining the outline of the scoring process in the state stage (4th stage) of a fertilized egg in the cell quality evaluation system of one Embodiment. It is a figure (the 2) explaining the outline of the scoring process in the state stage (4th stage) of a fertilized egg in the cell quality evaluation system of one Embodiment. It is a flowchart (the 1) which shows the process which is executed in the server apparatus of one Embodiment. It is a flowchart (the 2) which shows the process which is executed in the server apparatus of one Embodiment. It is a flowchart (the 3) which shows the process which is executed in the server apparatus of one Embodiment. It is a flowchart (the 4) which shows the process which is executed in the server apparatus of one Embodiment.

Hereinafter, embodiments according to the present invention will be described with reference to the drawings.

In the following embodiment, the cell quality evaluation system, the cell quality evaluation program, and the cell quality evaluation method of the present invention are used as a system for imaging the cells and evaluating the quality of the cells when culturing the cells. It is an embodiment when it is applied. However, the present invention is not limited to the following embodiments as long as the technical idea is included.

Further, in the following embodiment, in order to embody the explanation, in fertility treatment, a fertilized egg (embryo) obtained by fertilizing a sperm collected from a patient and an egg is cultured, and the superiority or inferiority of the quality of the fertilized egg is evaluated. The case will be described as an example.

[1] Outline of Cell Quality Evaluation System First, the configuration and outline of the cell quality evaluation system 1 in the present embodiment will be described with reference to FIGS. 1 and 2.

It should be noted that FIG. 1 is a system configuration diagram showing the configuration of the cell quality evaluation system 1 in the present embodiment, and FIG. 2 is a time after fertilization in a fertilized egg (embryo) fertilized with a sperm collected from a patient and an egg. It is a figure for demonstrating the development process of a fertilized egg with the course.

As shown in FIG. 1 of the cell quality evaluation system 1 of the present embodiment, a plurality of fertilized eggs E in culture are imaged, and an image of the imaged cell (hereinafter referred to as "cell image") is generated. It is a system for evaluating the superiority or inferiority of the quality of the fertilized egg E based on the generated cell image, and is a system for evaluating whether or not the requirements for transplantation to the mother's womb are satisfied by the score at each growth stage.

In particular, the cell quality evaluation system 1 of the present embodiment can also be used as a system for supporting the determination when the cell quality is finally determined by an operator such as a doctor.

For example, as shown in FIG. 2, in the fertilized egg E, the state stage indicating the internal state of the cell changes with the passage of time after the completion of fertilization.
(1) The first stage in which the plasma membrane, zona pellucida and polar body develop at least between 0 and 8 hours after fertilization.
(2) The second stage in which the protoplasmic membrane develops a pronucleus between 8 and 24 hours after fertilization.
(3) Within 20 to 100 hours after fertilization, the pronucleus disappears, cleavage occurs sequentially inside the zona pellucida, and the third stage in which morula develops, and
(4) The fourth stage of blastocyst development, in which the zona pellucida and plasma membrane disappear, and the cytotrophoblast and inner cell mass develop, 100 hours or more after fertilization.
It is designed to transition in the order of.

Therefore, the cell quality evaluation system 1 of the present embodiment is scored as a reference when evaluating the cell quality in each state stage by a score based on the cell image of each state stage and the cell image of each state stage. It has become possible to evaluate the superiority or inferiority of cell quality using reference information and.

Further, in general, in the state stages of each of these stages, the fertilized egg E transitions to a state having different internal structures such as a change in the number of cells, so that the same evaluation criteria and evaluation algorithms are used. It becomes difficult to accurately evaluate the quality of fertilized egg E in each stage by applying.

Then, in the fertilized egg E, the early stage of development after fertilization (that is, the first stage state stage (hereinafter, also simply referred to as “first stage”) and the second stage state stage (hereinafter, simply “second stage”). The quality of the fertilized egg in the "stage") is important for ensuring safety after transplantation, and the state stage from the fertilization to the third stage (hereinafter, simply referred to as "simply") including the first and second stages. The number of cells formed in a fertilized egg is the number of cells formed in the fertilized egg until the early and middle stages (also referred to as the "third stage") (the early state stage in which 2 egg splits occur and the middle state stage in which 4 egg splits occur). It has a great effect on quality.

On the other hand, when cleavage progresses from the latter stage of the third stage to the state stage including the fourth stage, the difference in the number of cells of about several cells has a small effect on the quality of the fertilized egg E. In particular, in the fourth stage, the number of cells contained in the inner cell mass increases to about 200 to 400, so even if the number of cells differs by several, the effect on the quality of the fertilized egg E is small. Become.

As described above, cells such as fertilized eggs change according to the state stage, and the quality determination criteria also change according to the state stage, so that it is difficult to uniformly evaluate the cells. Further, since the growth of cells is not uniform and the degree of growth changes for each individual cell, it may not be possible to accurately control quality while following the growth of each cell.

Therefore, as described above, since the internal state of the fertilized egg E is characteristically indicated by the number of cells and other states for each state stage, the cell quality evaluation system 1 of the present embodiment is a transition of each stage. Image the fertilized egg E to be evaluated as a cell image according to the timing, and perform quality evaluation in each period while changing the quality evaluation standard and quality evaluation algorithm according to each period to which the fertilized egg E belongs. In addition, it has a configuration for performing cell evaluation as a whole, and the quality of cells to be evaluated can be quantified, and the evaluation can be performed automatically and with high accuracy. It has become like.

The transition time of the state stage shown in FIG. 2 shows an example of the number of hours determined by a general rule of thumb, but the transition time is the culture environment of the fertilized egg E (for example, medium component, temperature, humidity). Etc.), so it is desirable to obtain the transition time in the actual culture environment by experiment in advance as a premise of this embodiment. However, in the present embodiment, the description will be given based on the above transition.

Specifically, as shown in FIG. 1, the cell quality evaluation system 1 of the present embodiment is cultured in each of a plurality of culture dishes (hereinafter referred to as “culture petri dishes”) 40-1 to 40-n. A camera 10 that captures images of fertilized eggs E1 to Ek (that is, fertilized egg E) of (k is, for example, 4) to generate a cell image, and a cell image that controls the camera 10 and is based on each state stage. It is equipped with a server device 30 that evaluates the quality of fertilized cells by using it.

The camera 10 is connected to the server device 30 directly or via a network (not shown), and images the corresponding cells at each state stage defined by the time axis to generate image data of the cell image. ..

In particular, the camera 10 captures the fertilized egg E cultured in the culture dish 40 at a predetermined timing based on the control from the server device 30 to generate a still image or a moving image, and the generated cell image. Is supplied to the server device 30 as image data.

In order to realize this function, the camera 10 is an optical system for adjusting the focus, zoom, and focus of the objective lens, flash, shutter, and lens, and a CCDI sensor (CCDI sensor) that converts an optical image input from the optical system into an electric signal. The Charge Coupled Device Image Sensor) unit, the generation unit that generates image data based on the electrical signal generated by the CCDI sensor unit, and the generation unit that generates image data based on the electrical signal generated by the CCDI sensor unit, and each unit is controlled according to the control signal from the server device 30, and the image of the fertilized egg E is displayed. It has a control unit for imaging.

The camera 10 may be connected to a microscope (not shown) by using an adapter or an attachment to capture a microscope image.

Further, in the present embodiment, as the imaging timing of each of the above state stages, for example, as illustrated in FIG. 2, "7 hours later", "20 hours later", "48 hours later", and "72", respectively, after insemination. It is set to "after an hour", "after 90 hours", "after 120 hours", etc., and the camera 10 is set to "after an hour", "after an hour", "after an hour", etc. The fertilized egg E to be evaluated is imaged (based on the elapsed time).

The server device 30 has various databases (hereinafter referred to as [DB]), and scores the quality of cells at each state stage according to the information recorded in the DB based on the cell image generated by the camera 10. Perform scoring evaluated by.

In particular, the server device 30
(1) Control the camera 10 and acquire the image data output from the camera 10.
(2) The scoring standard information, which is a standard for evaluating the quality of the fertilized egg E in each state stage based on the cell image of the image data, corresponds to the state stage identification information for identifying each state stage. Attach, control the recorded DB,
(3) For each state stage, while performing predetermined image processing on the cell image, a scoring process for scoring the quality of the corresponding fertilized egg E based on the corresponding scoring standard information is executed.
(4) An evaluation process for evaluating the superiority or inferiority of the quality of the fertilized egg E is executed according to the scoring result indicating the scored result.
(5) Present the evaluation result so that the user can view it.
It has a configuration.

Then, the server device 30 causes the camera 10 to image the fertilized egg E at different timings for each state stage to generate a cell image, and the server device 30 generates a cell image, and the corresponding fertilized egg is based on a plurality of cell images having different timings for each state stage. It has a configuration for scoring the quality of E.

Specifically, the server device 30 interlocks with the control of the camera 10 as the process of (3).
(3A) In order to evaluate the quality of the fertilized egg E in the first stage, the camera 10 is controlled about 7 hours after insemination to image the fertilized egg E in the first stage, and the cell image is analyzed. Score the quality of the first stage fertilized egg E based on the zona pellucida, progenitor membrane and polar body conditions.
(3B) In order to evaluate the quality of the fertilized egg E in the second stage, the camera 10 captures the image of the fertilized egg in the second stage about 20 hours after fertilization, and the cell image is analyzed to analyze the state of the pronucleus. Based on, score the quality of fertilized egg E in the second phase,
(3C) In order to evaluate the quality of fertilized eggs in the third stage, images of fertilized eggs in the third stage (early stage, middle stage and late stage) are displayed at each timing about 48 hours, 72 hours and 90 hours after fertilization. The number of blastomeres (2, 4 in principle) indicating the number of cells (blastomeres) divided by the fertilized egg E in the 3rd stage (early stage, middle stage and late stage) by analyzing the cell image while imaging with the camera 10. Score the quality of fertilized eggs in the third phase based on (8 or more)
(3D) In order to evaluate the quality of the fertilized egg E in the 4th stage, 120 hours after insemination, the camera 10 is made to take an image of the fertilized egg in the 4th stage, and the cell image is analyzed to analyze the trophoblast cells. Scoring the quality of fertilized eggs based on thickness and uniformity and inner cell mass size or cell number and clarity.
It has a configuration.

Further, the server device 30 has the processing of (4) as the process.
(4A) Based on the sum of the scores scored for each state stage, it is determined whether or not a predetermined condition (first condition) is satisfied, and if the condition is not satisfied, the above-mentioned As the superiority or inferiority of cell quality, the corresponding cell is judged to be unusable, or
(4B) It is determined whether or not each score in each state stage satisfies a predetermined condition (second condition) for each state stage, and if the condition is not satisfied, the corresponding cell. Is determined to be unusable,
It has a configuration.

For example, the server device 30 is adapted to determine whether or not the condition (4A) includes a condition such as a sum, average, or root mean square of the scores of all state stages, which is equal to or higher than a predetermined threshold value. .. In this case, for example, only the score at some state stages is high, the overall score is low, and cells that are not suitable for transplantation can be reliably excluded from the transplant target.

Further, the server device 30 is adapted to determine whether or not the score of each state stage satisfies a condition such as a predetermined threshold value or more as the condition (4B). In this case, the fertilized egg E, which is not highly evaluated in the most important initial state stage, is treated as a cell that is not suitable for transplantation even if the overall score is high and the transplantation requirement is satisfied. It can be definitely excluded.

The specific evaluation method of the fertilized egg E of the present embodiment is arbitrary, and for example, the sum, average, root mean square, etc. of each score scored for each state stage are taken to determine the value. Those exceeding the threshold value of are evaluated as being usable for transplantation, or the scores of each period are weighted according to the influence on the quality of fertilized egg E, and the final evaluation is performed. You may do so.

Further, when the evaluation is performed based on a part of the scores, for example, the final evaluation may be performed based on the scores of the first period and the second period, which have a great influence on the quality of the fertilized egg E. .. However, in the present embodiment, in order to make the explanation concrete, the explanation will be made assuming that the final evaluation is performed using the sum of the scores in all the state stages.

[2] Overall Configuration of Server Device [2.1] Next, the server device 30 of the present embodiment will be described with reference to FIGS. 3 to 6. 3 is a diagram showing the configuration of the server device 30 of the present embodiment, and FIGS. 4 to 6 are a cultured cell management DB 301, a state stage management DB 302, and scoring information provided in the server device 30, respectively. It is a figure which shows an example of the data recorded in DB 303.

As shown in FIG. 3, the server device 30 of the present embodiment includes a recording device 300 provided with various DBs 301 to 303, an interface unit 310, a ROM (Read only Memory) / RAM (Random Access Memory) 320, and the like. The display unit 330, the display control unit 340 that controls the image display on the display unit 330, the timer 360 for specifying the current time, the server management control unit 370 that integrally controls the entire server device 30, and the scoring process. It also has a data processing unit 380 for executing evaluation processing and the like, and an operation unit 390.

Each of these parts is interconnected by the bus B, and data is exchanged between the components via the bus B.

The recording device 300 is composed of an HDD (Hard Disk Drive) or an SSD (Solid State Drive). The recording device 300 has a cultured cell management DB 301, a state stage management DB 302, and a scoring information DB 303.

For example, the scoring information DB 303 of the present embodiment constitutes the recording means of the present invention.

The cultured cell management DB 301 is a DB in which information for managing the fertilized egg E during culture (hereinafter referred to as “fertilized egg management information”) is recorded as data in the culture dish 40 of the cell quality evaluation system 1. Is. For example, in the cultured cell management DB 301, as shown in FIG. 4,
(1) A culture dish ID for identifying each culture dish 40,
(2) A fertilized egg ID for identifying each fertilized egg E in culture, and
(3) The date and time when each fertilized egg E was fertilized (hereinafter referred to as "fertilization date and time"),
(4) Each score calculated by the scoring process in each state stage of each fertilized egg E and
(5) A total score showing the scoring results in all state stages and
Fields for recording are provided in association with each other.

The culture petri dish ID, fertilized egg ID, and fertilization date and time information recorded in the cultured cell management DB 301 are registered in advance by the user when the fertilized egg E is fertilized and the culture is started. Further, in the score of each state stage of (4), information indicating an error (for example, a value indicating an error) is recorded when the score cannot be calculated and an error occurs as described later.

For example, in FIG. 4, in a culture dish 40 having a “D001” culture dish ID, four fertilized eggs E corresponding to the fertilized egg IDs of “E001 to 004” are cultured, and each fertilized egg E is in a state stage. And the total score through all state stages (in this embodiment, the sum of the scores) indicate the fertilized egg management information recorded in association with each other.

In particular, in FIG. 4, the fertilization date and time is "13:00 on May 10, 2014", the score of the first period is "0 points", the score of the second period is "10 points", and the score of the third period is "10 points". The score is "12 points", the score of the 4th period is "8 points", and the fertilized egg management information of the cells of "E001" whose total score is "30 points" is recorded. An example is shown.

Note that FIG. 4 shows a recording example of information when four fertilized eggs E are cultured in each culture dish 40, but the cells to be cultured in one culture dish 40 are 4 cells. When the number of fertilized eggs is 4 or more, the number of fertilized eggs is not limited to four, and a data recording field may be added to the cultured cell management DB 301 as needed.

As for the score of the third period, the comprehensive score in a plurality of periods of the first half, the middle and the second half is recorded.

The state stage management DB 302 is a DB for recording information for managing each state stage of the first to fourth stages (hereinafter, referred to as “state stage management information”) generated in the fertilized egg E as data. Is.

For example, in the state stage management DB 302, as shown in FIG.
(1) A state stage ID for identifying each state stage of the fertilized egg E, and
(2) The state stage name corresponding to the state stage ID and
(3) Imaging timing for imaging the fertilized egg E in the relevant state stage, and
Are recorded in association with each other.

In addition, in FIG. 5, the state stage IDs “S001” to “S006” assigned to each of the first to fourth periods and the state stage names “1st period” to “4th” corresponding to each ID are shown. An example is shown in which the "period" is recorded, and as the imaging timing of each state stage, the times one hour before and one hour after the transition timing in the state stage are recorded as the imaging timing.

In particular, FIG. 5 shows an example in which three timings of “6, 7 and 8 hours later” are recorded as imaging timings for the “first phase” of “S001”.

The scoring information DB 303 is a DB in which scoring information used when executing the scoring process is recorded. For example, as shown in FIG. 6, the scoring information DB 303 is information necessary for scoring processing in each state stage in association with the state stage ID (that is, for each state stage), and the scoring is performed. Judgment criteria for scoring in the process, scoring criteria information indicating the scoring criteria, and scoring information including an algorithm for performing the scoring process are recorded.

Specifically, the scoring information in the first period of this embodiment includes
(1A) Judgment for determining the presence or absence of the protoplasm, the presence or absence of the zona pellucida, the presence of the zona pellucida and the protoplasmic membrane, and the presence or absence of the polar body generated between the transparent body and the protoplasmic membrane. Standards and
(1B) Score values for each criterion and
(1C) Score correction values at multiple timings and
Is included.

In addition, as the score value included in the scoring information of the present embodiment,
(1B-1) Score in the absence of the zona pellucida (1B-2) Score in the absence of the zona pellucida (1B-3) There is a zona pellucida and the zona pellucida, and between the zona pellucida and the zona pellucida. Score when there is no generated polar body (1B-4) Number of polar bodies when there is a zona pellucida and a progenitor membrane and there is a polar body generated between the transparent body and the progenitor membrane. Score (1B-5) when there is one zona pellucida and the original plasma membrane, and when there is a polar body generated between the transparent body and the original plasma membrane, the number of polar bodies is 2. Score in the case of (1B-6) When there is a zona pellucida and a progenitor membrane and there is a polar body generated between the transparent body and the progenitor membrane, the number of polar bodies is 3 or more. The score in the case of is used.

For example, the scoring information of the present embodiment includes scoring reference information corresponding to the state stage of the "first stage".
・ Score when there is no zona pellucida: 0 points ・ Score when there is no zona pellucida: 0 points ・ There is a zona pellucida and a zona pellucida, and there is no polar body generated between the zona pellucida and the zona pellucida. Case (case3) score: 0 points ・ When the zona pellucida and the original plasma membrane are present and there is a polar body generated between the transparent body and the original plasma membrane, the number of polar bodies is one. Score: 6 points-Score when there are zona pellucida and protozoal membrane, and there are polar bodies generated between the transparent body and the progenitor membrane, and the number of polar bodies is 2. : 10 points ・ Score when the number of polar bodies is 3 or more in the presence of the zona pellucida and the original plasma membrane and the polar bodies generated between the transparent body and the original plasma membrane: 0 The points are memorized.

Further, as the correction value of the score at the plurality of timings in (1C), a value indicating the rate of increase / decrease of the above score value is recorded. For example, if the standard timing of the first period is 7 hours later and 1 hour before and after the standard timing (that is, 6 hours and 8 hours later) is defined as the imaging timing, the score is corrected at a plurality of timings. As a value, "0%" is recorded after 7 hours, "20%" after 6 hours, and "-20%" after 8 hours in each score.

In the present embodiment, the score value is corrected according to the imaging timing, for example, "12 points" when two polar bodies occur after 6 hours and "8 points" when they occur after 8 hours (8 points). By increasing or decreasing), the score of the fertilized egg E with good growth can be set high, and the fertilized egg E with a high growth rate can be surely selected as the transplant target.

The scoring information in the second phase of this embodiment includes
(2A) Criteria for determining the presence or absence of pronucleus inside the plasma membrane,
(2B) Score values for each criterion and
(2C) Score correction values at multiple timings and
Is included.

In addition, as the score value included in the scoring information of the present embodiment,
(2B-1) Score when there is no pronucleus inside the plasma membrane (2B-2) Score when there is a pronucleus inside the plasma membrane and the number of the pronuclei is one (2B-1) -3) Score when there is a pronucleus inside the plasma membrane and the number of the pronuclei is 2. (2B-4) When there is a pronucleus inside the plasma membrane The score when the number of nuclei is 3 or more is used.

For example, the scoring information of the present embodiment includes scoring reference information corresponding to the state stage of the "second phase".
-Score when there is no protoplasmic membrane inside: 0 points-Score when there is a protoplasmic membrane inside and the number of protoplasmic membranes is 1.-Protoplasmic membrane Score when there are pronuclei inside and the number of pronuclei is 2. 10 points ・ When there are pronuclei inside the plasma membrane and the number of pronuclei is 3 or more Score in the case of: -10 points are stored.

Further, as the correction value of the score at the plurality of timings in (2C), a value indicating the rate of increase / decrease of the above score value is recorded as in the first period. For example, if the standard timing of the second period is 20 hours later and 1 hour before and after the standard timing (that is, 19 hours and 21 hours later) is specified as the imaging timing, the score is corrected at a plurality of timings. As a value, "0%" is recorded after 20 hours, "20%" after 19 hours, and "-20%" after 21 hours in each score.

In the present embodiment, the score value is determined according to the imaging timing, for example, "12 points" when two pronuclei occur after 19 hours, or "8 points" when two pronuclei occur after 21 hours. By correcting (increasing or decreasing), the score of the fertilized egg E with good growth can be set high, and the fertilized egg E with a high growth rate can be surely selected as the transplant target.

The scoring information in the first half, middle and second half of the third period of the present embodiment includes
(3A) Judgment criteria for determining the presence or absence of blastomere and
(3B) The value of the score against the judgment criteria and
(3C) Score correction values at multiple timings and
Is included.

In addition, as the score value included in the scoring information of the present embodiment,
(3B-1) Score when there is no blast ball (3B-2) Score when the number of blast balls is less than the appropriate number (3B-3) Score when the number of blast balls is appropriate (3B-4) Score when the number of blast balls is appropriate (3B-4) The score is used when is greater than the appropriate number.

For example, the scoring information of the present embodiment includes scoring reference information corresponding to the state stage of the "third period".
-Score when there is no blast ball: -10 points-Score when the number of blast balls is a predetermined ratio (for example, 75%) from the appropriate number: 8 points-The number of blast balls is the appropriate number (for example) Score: 10 points for the case of 2 in the first half, 4 in the middle and 8 in the second half) ・ Score: 12 points when the number of blastomeres is more than the appropriate number is stored.

Further, as the correction value of the score at the plurality of timings in (3C), a value indicating the rate of increase / decrease of the above score value is recorded as in the first period and the second period. For example, the standard timings of the first half, middle and second half of the third period are 48 hours, 72 hours and 90 hours, and 1 hour before and after the standard timing (that is, 47 hours and 49 hours in the case of the first half). , 71 hours and 73 hours in the case of the middle term, and 89 hours and 91 hours in the case of the latter term) is specified as the imaging timing, and the score is corrected at a plurality of timings. As a value, in the case of the first half, "0%" is recorded after 48 hours, "20%" after 47 hours, and "-20%" after 49 hours. The same figures have been recorded in the middle and late stages.

In the present embodiment, for example, in the case of the previous period, when two blastomeres occur after 47 hours, "12 points", or when they occur after 49 hours, "8 points", and so on. By correcting (increasing or decreasing) the score value by, the score of the fertilized egg E with good growth can be set high, and the fertilized egg E with a fast growth rate can be surely selected as the transplant target. It has become.

The scoring information in the fourth period of this embodiment includes
(4A) Criteria for determining the uniformity of trophoblast cell thickness, trophoblast cell thickness, inner cell mass size, inner cell mass clarity, and timing when predetermined criteria are met, and
(4B) The value of the score against the judgment criteria and
(4C) Score correction values at multiple timings and
Is included.

In addition, as the score value included in the scoring information of the present embodiment,
(4B-1) Scores of each grade of 3 grades (excellent, good, acceptable) in the thickness of trophoblast cells, and
(4B-2) Scores of each grade of 3 grades (excellent, good, acceptable) in the size of the inner cell mass are used.

For example, the scoring information of the present embodiment includes scoring reference information corresponding to the state stage of the "fourth period".
-Score when the thickness of the vegetative membrane cells is assumed to be uniform and the number of cells is a predetermined number or more: 5 points-The thickness of the vegetative membrane cells is assumed to be uniform and the number of cells is a predetermined number Score when less than: 3 points ・ Score when the thickness of the nutrient membrane cells is outside the range assumed to be uniform: 0 points ・ The size of the inner cell mass and the clarity on the image are each equal to or higher than the predetermined thresholds. Score: 5 points-Score when the size of the inner cell mass is less than the predetermined threshold and the clarity on the image is greater than or equal to the predetermined threshold: 5 points-The size of the inner cell mass is predetermined Score when it is above the threshold and the clarity on the image is less than the predetermined threshold: 3 points ・ Score when the size of the inner cell mass and the clarity on the image are less than the predetermined threshold: 0 points Is remembered.

Further, as the correction value of the score at the plurality of timings in (4C), a value indicating the rate of increase / decrease of the above score value is recorded as in the first to third periods. For example, if the standard timing of the 4th period is 90 hours later and 1 hour before and after the standard timing (that is, 89 hours and 91 hours later) is defined as the imaging timing, the score is corrected at a plurality of timings. As a value, "0%" is recorded after 90 hours, "20%" after 89 hours, and "-20%" after 49 hours in each score.

In the present embodiment, for example, after 90 hours have passed, the thickness of the trophoblast cells is within the range assumed to be uniform and the number of cells is a predetermined number or more, and the size of the inner cell mass and the image. "12 points" is used as a score when the intelligibility is equal to or higher than a predetermined threshold value.

The interface unit 310 is for connecting to the camera 10 by wire or wirelessly using a communication standard such as USB (Universal Serial Bus), Ethernet (registered trademark), or IEEE802.11 system wireless LAN (Loacal Area Network). It has a function.

Then, the interface unit 310 transmits a control signal for controlling the camera 10 by communicating with the connected camera 10, and also receives the image data output from the camera 10.

For example, the interface unit 310 of the present embodiment constitutes the acquisition means of the present invention together with the data processing unit 380.

Various programs necessary for driving the server device 30 are recorded in the ROM / RAM 320. The ROM / RAM 320 is also used as a work area when various processes are executed.

The display unit 330 is composed of a panel of a liquid crystal element or an EL (Electroluminescence) element, and displays a predetermined image based on the display data generated by the display control unit 340. For example, the display unit 330 of the present embodiment constitutes the presentation means of the present invention together with the display control unit 340.

Under the control of the server management control unit 370 and the data processing unit 380, the display control unit 340 generates drawing data necessary for the display unit 330 to draw a predetermined image, and the generated drawing data is used as the display unit 330. Output to. For example, the display control unit 340 of the present embodiment, in conjunction with the data processing unit 380, constitutes the presentation means of the present invention together with the display unit 330.

The timer 360 is used to manage the time required to manage each state stage.

The server management control unit 370 is mainly composed of a central processing unit (CPU), and integrates and controls each part of the server device 30 by executing a program.

The data processing unit 380 is configured by an independent central processing unit (CPU), or is configured by using the central processing unit (CPU) of the server management control unit 370.

In particular, the data processing unit 380 manages the imaging of the fertilized egg E by the camera 10 by executing the program and the algorithm based on the information recorded in each DB 301 to 303 under the control of the server management control unit 370. The imaging management unit 381, the cultured cell management unit 382 for managing each fertilized egg E cultured in the culture chalet 40, and the first scoring process corresponding to each of the first to fourth stages are executed. The scoring processing unit 383, the second scoring processing unit 384, the third scoring processing unit 385, the fourth scoring processing unit 386, and the fertilized egg evaluation unit 387 that executes the evaluation processing of the fertilized egg E are realized. ..

For example, the imaging management unit 381 and the cultured cell management unit 382 of the present embodiment, in conjunction with the camera 10, constitute the imaging control means and the acquisition means of the present invention, and the first scoring processing units 383 to 4 The scoring processing unit 386 constitutes the scoring means of the present invention. Further, for example, the fertilized egg evaluation unit 387 of the present embodiment constitutes the evaluation means of the present invention.

The image pickup management unit 381 outputs a control signal including setting values such as a shooting angle, a shutter speed, an exposure value, and an aperture to the camera 10 in order to capture an image of the fertilized egg E to be evaluated by the camera 10. The image data of the cell image output from 10 is acquired. Then, the image pickup management unit 381 records the acquired image data in the ROM / RAM 320 as data for scoring processing.

Specifically, the imaging control unit 381 controls the camera 10 mounted on the installation table having the angle changing function provided facing the culture dish 40 together with the installation table, and the predetermined control By outputting the signal to the camera 10, the shooting angle of the camera 10 is adjusted.

In addition, the imaging management unit 381 determines the imaging timing according to the fertilization date and time recorded in the cultured cell management DB 301, the imaging timing recorded in the state stage management DB 302, and the current date and time acquired by the timer 360, and also determines the imaging timing. A predetermined control signal is output to the camera 10 at the imaging timing, and the camera 10 is made to image the fertilized egg E.

The first scoring processing unit 383 to the fourth scoring processing unit 386 perform scoring processing based on the cell image of the fertilized egg E in the corresponding state stage, respectively.

In particular, the first scoring processing unit 383 to the fourth scoring processing unit 386 are applied to the scoring of the corresponding state stage after the camera 10 acquires the image data in the corresponding state stage or at a predetermined timing. The scoring information to be sought is read from the scoring information DB 303, and the scoring process for scoring in each state stage is executed using the read scoring information.

The details of each scoring process of the first scoring processing unit 383 to the fourth scoring processing unit 386 in this embodiment will be described later.

The fertilized egg evaluation unit 387 executes an evaluation process based on the scores of each state stage recorded in the cultured cell management DB 301, or an evaluation process of evaluating the fertilized egg E based on the sum of the scores of all the state stages. Then, the result is recorded in the cultured cell management DB 301 to update the cultured cell management DB 301.

In addition, the fertilized egg evaluation unit 387 evaluates whether or not each fertilized egg is suitable for transplantation into the mother's womb by comparing the overall score with a predetermined first threshold value (first condition). The evaluation process is executed, and the evaluation result is displayed on the display unit 330 so that the user can present it. That is, the fertilized egg evaluation unit 387 evaluates that the fertilized egg cannot be used for transplantation when the overall score is lower than the first threshold value, and evaluates that the fertilized egg can be transplanted when it is equal to or higher than the first threshold value.

For example, when a plurality of fertilized eggs E are cultured and the same process is sequentially repeated for each fertilized egg E to perform scoring, the total score corresponding to each fertilized egg E is stored in the cultured cell management DB 301. Is stored, the fertilized egg evaluation unit 387 may sort the fertilized eggs E having the highest overall score in order and arrange them in descending order of the overall score and display them on the display unit 330. In this case, the user can freely select a fertilized egg E that can be used for transplantation and use it for transplantation into the mother's womb.

The operation unit 390 is a large number of keys such as various buttons, operation buttons for inputting each operation command, and a numeric keypad, and is composed of a touch sensor, a keyboard, and a mouse provided on the display unit 330 to perform each operation. It has come to be used in some cases.

[2.2] Scoring process Next, using FIGS. 7 to 15, the first scoring processing unit 383, the second scoring processing unit 384, the third scoring processing unit 385, and the fourth score of the present embodiment are used. Each scoring process executed by the ring processing unit 386 will be described.

[2.2.1] First-stage scoring process First, the first-stage scoring process executed by the first scoring processing unit 383 of the present embodiment will be described with reference to FIGS. 7 and 8. 7 and 8 are diagrams for explaining the scoring process in the first period executed by the first scoring process unit 383.

The first scoring processing unit 383 acquires the scoring information (including the scoring reference information and the algorithm) in the first period recorded in the scoring information DB 303, and the cell image acquired by the imaging management unit 381. Therefore, while performing predetermined image processing on the cell image of the first stage of the state stage, scoring of the fertilized egg E in the cell image is executed.

Specifically, the first scoring processing unit 383 uses the cell image in the state stage of the first stage, that is, the cell image at the time when 6 hours, 7 hours, and 8 hours have passed since fertilization, as the cell image. ..

Then, the first scoring processing unit 383 performs the predetermined image processing of the object imaged by detecting the pixels in which the color feature amount such as the brightness changes discontinuously in each cell image. The edge detection process for detecting the edge and the threshold process for generating a mask image of the plasma membrane by binarizing with a predetermined threshold are executed, and the plasma membrane in the fertilized egg E imaged in the cell image is transparent. Detects body and polar body.

In addition, the first scoring processing unit 383 scores each cell image according to the presence / absence, position, number, etc. of the plasma membrane, the transparent body, and the polar body, and a plurality of cells such as the mean or the sum in each cell image. Determine the score for the first phase using images.

For example, in the case of the cell image of the fertilized egg E in the state stage of the first stage as shown in FIG. 7 (A), the first scoring processing unit 383 performs the edge detection processing (FIG. 5) based on the preconditions. [1-1]) to generate an edge image (see FIG. 7B), and threshold processing ([1-2] in FIG. 5) to perform a mask image of the plasma membrane (FIG. 7). 7 (C)) is generated.

The precondition is a condition for detecting the plasma membrane and the polar body, that is,
(A) A parameter (hereinafter referred to as "parameter") OR, which indicates in advance the minimum to maximum value (OR (min) to OR (max)) of the radius of the plasma membrane.
(B) Parameters indicating the values (PR1 (min) to PR1 (max)) from the minimum value to the maximum value of the polar body radius (hereinafter referred to as "parameters") PR1 and (C) the plasma membrane are cell images. Condition that it exists near the center of
In this embodiment, each numerical value and condition shall be specified in the algorithm.

Further, since the edge detection process is the same as the conventional one, the details thereof will be omitted. Further, in the threshold processing, for example, a threshold value may be set in advance for the brightness value of the pixel, and a binary image in which the pixel whose brightness is equal to or less than the threshold value is black may be generated.

Further, if the plasma membrane, the polar body, or both of them cannot be detected based on the above preconditions, the first scoring processing unit 383 indicates an error for the corresponding cell image as an error processing. Information (for example, information indicating an error) is registered in the state stage management DB 302.

On the other hand, when the first scoring processing unit 383 can detect both the plasma membrane and the polar body, the first scoring processing unit 383 applies a mask image of the plasma membrane from the edge image obtained by the polar body edge detection processing to perform an edge. A removal process ([1-3] in FIGS. 7 and 8) for removing the edge portion of the zona pellucida from the image is executed, and an edge image as shown in FIG. 8 (A) (hereinafter, “protoplasmic membrane edge image”” is executed. ) Is generated.

In the present embodiment, if the circle detection such as the Hough transform is simply executed on the edge image, a lot of noise may be generated. Therefore, in order to limit the search range in the circle detection in advance, the first score The ring processing unit 383 detects the plasma membrane, extracts the edges around the boundary of the mask image, and generates the plasma membrane edge image excluding the transparent band.

Then, the first scoring processing unit 383 executes a Hough conversion as a circle detection on the plasma membrane edge image based on the parameter OR indicating the radius of the plasma membrane ([1-4] in FIG. 8). An edge image of only the polar body in which the edge of the plasma membrane included in the plasma membrane edge image is detected and the edge of the plasma membrane is excluded from the plasma membrane edge image (hereinafter, "polar body edge image (FIG. 8)". (Refer to (B)) ”).

Further, as shown in FIG. 8C, the first scoring processing unit 383 executes a Hough transform as a circle detection on the polar body edge image based on the polar body radius parameter PR1 (FIG. 8). [1-5]), the polar body is extracted while removing noise.

On the other hand, when the polar body is extracted, the first scoring processing unit 383 determines the score of the first period based on the number of the extracted polar bodies.

In particular, the first scoring processing unit 383 normalizes the state in which two polar bodies, an egg-derived polar body and a sperm-derived polar body, are present.
(A) If there are two polar bodies, "10 points",
(B) If there are three polar bodies, "0 point",
(C) If there is only one polar body, "5 points" and
(D) When there are 0 polar bodies, "0 point",
And score.

In the case of (B) above, it is determined that it is not normal as it is a multi-sperm fertilization in which two sperms are fertilized in one egg, and in the case of (C) above, insemination fails. Since there is a possibility that the polar bodies may overlap each other depending on the imaging angle, the above scores are given.

Further, the case of (D) above includes, for example, a case where an error occurs when generating an edge image or a mask image of the plasma membrane.

[2.2.2] Second Phase Scoring Process First, the second phase scoring process executed by the second scoring processing unit 384 of the present embodiment will be described with reference to FIGS. 9 and 10. 9 and 10 are diagrams for explaining the scoring process in the second period executed by the second scoring process unit 384.

The second scoring processing unit 384 acquires the scoring information (including the scoring reference information and the algorithm) in the second period recorded in the scoring information DB 303, and is the imaging management unit as in the first period. The cell image acquired by 381, in which the state stage performs predetermined image processing on the cell image of the second stage, the scoring of the fertilized egg E in the cell image is performed.

Specifically, the second scoring processing unit 384 uses an image of a fertilized egg E (or a fertilized egg E presumed to be a pronuclear stage embryo) that has become a pronuclear stage embryo, and also uses cells. As the image, the cell image in the state stage of the second stage, that is, the cell image at the time when 19 hours, 20 hours and 21 hours have passed from fertilization is used.

Then, the second scoring processing unit 384 performs the predetermined image processing of the object imaged by detecting the pixels in which the color feature amount such as the brightness changes discontinuously in each cell image. An edge detection process for detecting an edge and a threshold process for generating a mask image of the plasma membrane by binarizing with a predetermined threshold are executed, and the plasma membrane and polar body in the fertilized egg E imaged in the cell image are executed. Detect the body and protoplasm.

In addition, the second scoring processing unit 384 performs scoring in each cell image according to the presence / absence, position, number, etc. of the plasma membrane, polar body, and pronucleus, and a plurality of cells such as mean or sum in each cell image. Determine the score for the second phase using images.

For example, in the case of the cell image of the fertilized egg E in the state stage of the second stage as shown in FIG. 9A, the second scoring processing unit 384 is based on the preconditions as in the first stage. , Edge detection processing ([2-1] in FIG. 9) is executed to generate an edge image (see FIG. 9B), and threshold processing ([2-2] in FIG. 9) is executed to generate the original image. A mask image of the plasma membrane is generated as a first mask image (see FIG. 9C).

Then, unlike the first stage, the second scoring processing unit 384 executes a filter processing using a non-linear filter such as a morphological processing on the first mask image due to the positional characteristics of the plasma membrane. The degeneracy process is executed ([2-3] in FIG. 9) to generate a second mask image (see FIG. 9D).

In addition to the first precondition, the preconditions for the second period include parameters (hereinafter, "parameters") indicating values (PR2 (min) to PR2 (max)) from the minimum value to the maximum value of the pronuclear radius. In this embodiment, each numerical value and condition shall be specified in the algorithm, including PR2.

Further, for example, since the plasma membrane often exists in the center of the cell and rarely touches the vicinity of the boundary, the second scoring processing unit 384 generates the second mask image by the above-mentioned filter processing. doing.

Further, if the plasma membrane, the polar body, the pronucleus, or none of them can be detected, the second scoring processing unit 384 performs error processing on the corresponding cell image based on the above preconditions. Information indicating an error (for example, information indicating an error) is registered in the state stage management DB 302.

On the other hand, the second scoring processing unit 384 applies the second mask image of the plasma membrane from the edge image obtained by the edge detection processing when any of the plasma membrane, the polar body and the pronucleus can be detected. Then, as in the first phase, a removal process ([2-4] in FIGS. 9 and 10) for removing the edge portion of the transparent band from the edge image is executed, and the protoplasm as shown in FIG. 10 (A) is executed. Generate a film edge image.

In this embodiment, if circle detection such as Hough transform is simply performed on the edge image, a lot of noise may be generated. Therefore, the search range for detecting the pronucleus existing in the plasma membrane is set in advance. In order to limit to, the plasma membrane is detected, the inside of the protoplasmic membrane second mask image is extracted, and the plasma membrane edge image excluding the transparent band is generated.

Then, the second scoring processing unit 384 executes a Hough conversion (circle conversion) as a circle detection on the plasma membrane edge image based on the parameter OR indicating the radius of the plasma membrane ([2-] in FIG. 10]. 5]), an edge image of only the protoplasmic membrane in which the edge of the plasma membrane included in the plasma membrane edge image is detected and the edge of the plasma membrane is excluded from the plasma membrane edge image (hereinafter, "protoplasmic membrane edge"). An image (referred to as "FIG. 10 (B))") is generated.

Further, the second scoring processing unit 384 executes a Hough transform as a circle detection on the previous edge image based on the polar body radius parameter PR1 and the pronucleus radius parameter PR2 to remove the pronucleus while removing noise. Extract ([2-6] in FIG. 10).

On the other hand, when the polar body and the pronucleus are extracted, the second scoring processing unit 384 determines the score of the second period based on the number of the extracted polar bodies and the pronuclei.

In particular, the second scoring processing unit 384 is fertilized normally, and one each is generated from the egg-derived pronucleus and polar body and the sperm-derived pronucleus and polar body (that is, the polar body and the polar body). 2 each, 4 in total), assuming that the existence is normal
(A) When there are two polar bodies and two pronuclei (that is, when there are four), "10 points",
(B) If there are two polar bodies and one pronucleus, "5 points",
(C) When there is one polar body and three pronuclei, "-10 points" and
(D) In other cases, "0 points",
And score.

In the case of (B) above, not only the fertilization has failed, but also the fertilization has failed, and there is a possibility that the polar bodies are overlapped with each other depending on the imaging angle. Therefore, the above scores are given.

In addition, if the number of polar bodies and pronuclei generated is actually only one, the score in the later state stage will be low, so even if a score of about "5 points" is given as described above. , It is possible to judge the quality of the fertilized egg E from the score throughout the whole period.

[2.2.3] Third-stage scoring process Next, the third-stage scoring process executed by the third scoring processing unit 385 of the present embodiment will be described with reference to FIGS. 11 to 13. .. 11 to 13 are diagrams for explaining the scoring process in the third period executed by the third scoring process unit 385.

In the scoring process of the third period of the present embodiment, as will be described later, the third period will be described by dividing it into three state stages, but the third scoring processing unit 385 has a score in one state stage. The ring process may be performed, or the scoring process in 4 or more state stages may be performed, and the number of state stages is determined based on the type of cell to be evaluated and other environmental conditions.

The third scoring processing unit 385 acquires the scoring information (including the scoring reference information and the algorithm) in the third period recorded in the scoring information DB 303, and is the same as in the first and second periods. , The cell image acquired by the imaging control unit 381, and the state stage performs the scoring of the fertilized egg E in the cell image while executing the predetermined image processing on the cell image in the third stage.

In particular, the third scoring processing unit 385 divides the third period into a plurality of stages, divides each of the divided stages (hereinafter, referred to as "state division stage"), and executes predetermined image processing. Scoring of the fertilized egg E in the cell image is performed.

Specifically, the third scoring processing unit 385 is a fertilized egg E (or a fertilized egg presumed to be these embryos) that is within the 2-cell stage embryo, 4-cell stage embryo, and 8-cell stage. In addition to using the image of E), as described above, the divided state stage is divided into three divided state stages as the early stage, the middle stage, and the late stage, and the cell image is the cell image in each divided state stage of the third stage. That is, in the case of the early stage, the cell image at the time when 47 hours, 48 hours and 49 hours have passed since the fertilization, and in the case of the middle stage, the cell image at the time when 71 hours, 72 hours and 73 hours have passed since the fertilization. And, in the case of the late stage, the cell image at the time when 89 hours, 90 hours and 91 hours have passed since the fertilization is used.

Then, as predetermined image processing, the third scoring processing unit 385 includes contour detection processing for detecting the contour in the zona pellucida in each cell image for each of the prophase, metaphase, and anaphase divided state stages. The concave point detection process for detecting the number of concave points of the detected contour is executed, and the number of cells formed in the zona pellucida in the fertilized egg E imaged in the cell image is detected.

In addition, the third scoring processing unit 385 performs scoring in each cell image according to the number of cells, that is, the number of blastomeres, and also uses a plurality of cell images such as the average or the sum of each cell image in the third phase. The score in each divided state stage is determined, and the score for the entire third period is calculated by the average or the sum of the scores in the first half, the middle and the second half.

In the scoring process of the third stage of the present embodiment, the number of cells to be detected in the first stage, the middle stage, and the second stage is only 2, 4, and 8, respectively, and the score in the first stage is different. The ring processing will be described, and the description of the scoring process in the middle and late stages will be omitted.

For example, in the case of the cell image of the fertilized egg E in the state stage of the third stage as shown in FIG. 11 (A), the third scoring processing unit 385
(1) An edge extraction image (see FIG. 11B) is generated by performing an edge detection process ([3-1] in FIG. 11) in the same manner as in the first and second phases.
(2) Short small edges having a predetermined length or less are deleted as noise from the edge extracted image ([3-2] in FIG. 11) to generate a small edge deleted image (see FIG. 11 (C)).
(3) Small edge deletion The edge of an unnecessary region is deleted from the image by using a predetermined mask image (mask image for detecting a region within a certain range from the center of the cell image) (FIG. 11). [3-3]), a main edge image (see FIG. 11 (D)) obtained by deleting edges having a length equal to or less than a predetermined length and extracting the main edge is generated.
(4) A binarization process in which a threshold value for binarization with the main edge as a boundary is set and the edge extracted image is binarized based on the set threshold value ([3-4] in FIGS. 11 and 12). To generate a binarized image (see FIG. 12 (A)).
(5) Correction processing such as degeneracy processing and expansion processing for forcibly binarizing the inner pixel and the outer pixel in contact with the main edge of the generated binarized image ([FIG. 12] 3-5]) is performed to generate a corrected image (see FIG. 12B).
(6) The binarized boundary line of the generated corrected image is detected as a contour ([3-5] in FIG. 12).
Each process of the contour detection process is executed to generate a contour image (see FIG. 12C).

On the other hand, when the contour image is generated, the third scoring processing unit 385 calculates the curvature for each predetermined length with respect to the contour, and as shown in FIGS. 13A and 13B, the value is negative. The number of values that becomes is detected as the number of cells. That is, in the fertilized egg E, the number of concave points is proportional to the number of blastomeres. If the number of concave points is 2, the number of blastomeres is "2", and if the number of concave points is 3, the number of blastomeres is "4". Therefore, the third scoring processing unit 385 can specify the number of blastomeres generated according to the number of concave points.

In FIGS. 13A and 13B, the horizontal axis represents the distance from the starting point of the curvature calculation, and the vertical axis represents the curvature. Further, in the example of FIG. 13, since two concave points are shown, the number of cells is "2" in this case.

Further, when detecting the number of cells, the third scoring processing unit 385 divides the cell portion assumed from the number of concave points in order to determine the uniformity of the size in the cell portion of the cell image based on the detected number of concave points. An ellipse that corrects the ellipse of the divided area by using edges other than the main edge detected by the edge detection process (for example, those deleted by the edge near the main edge) while executing the segmentation process to determine the area. The number of cells may be corrected while the fitting process is executed to correct the contour image.

On the other hand, when the third scoring processing unit 385 detects the number of cells, the score of the third stage is determined based on the detected number of cells.

In particular, the third scoring processing unit 385 considers that it is normal to have two blastomeres in the case of the previous term.
(A) 6 points when there is one specified number of blastomeres (B) 10 points when there are two specified numbers of blastomeres,
(C) If there are 3 specified blastomeres, 12 points,
(D) If there are four or more specified blastomeres, scoring is performed with 0 points.

In addition, in the case of the above (A), there is a possibility that the growth is slow, and a score is given in consideration of that case, and in the case of the above (C), there is a possibility that the growth is fast. A score is given in consideration of the case, and in the case of 4 or more, there is a high possibility that the size of the blastomeres is uneven, and there is a high possibility that the grade is low. It is attached.

Further, the case of (D) above includes, for example, a case where an error occurs during the edge detection process or the concave point detection process.

Further, the third scoring processing unit 385 considers that it is normal for the number of blastomeres to be 4 and 8 in each of the middle and late stages, respectively.
(A) When the number of specified blasts is 2 or less, or 6 and 7 or less, 6 points (B) When the number of specified blasts is 4 or 8 Is 10 points,
(C) If the specified number of blastomeres is 8 or 16, 12 points,
(D) If the specified number of blastomeres is 10 or more, or 20 or more, the score is set to 0 points.

[2.2.4] Fourth Phase Scoring Process Next, the fourth phase scoring process executed by the fourth scoring processing unit 386 of the present embodiment will be described with reference to FIGS. 14 and 15. 14 and 15 are diagrams for explaining the scoring process in the fourth period executed by the fourth scoring process unit 386.

The fourth scoring processing unit 386 acquires the scoring information (including the scoring reference information and the algorithm) in the fourth period recorded in the scoring information DB 303, and is the same as in the first to third periods. , The cell image acquired by the imaging control unit 381, and the state stage performs the scoring of the fertilized egg E in the cell image while executing the predetermined image processing on the cell image in the fourth stage.

Specifically, the fourth scoring processing unit 386 is a fertilized egg E (or a blastocyst stage embryo (transplanted embryo)) that has become a blastocyst stage embryo (transferred embryo). The image of the embryo E) is used, and as the cell image, the cell image in the state stage of the fourth stage, that is, the cell image at the time when 199 hours, 120 hours, and 121 hours have passed from the fertilization is used.

Then, the fourth scoring processing unit 386 performs image processing for determining the trophoblast cells formed in the blastocyst shown in FIG. 2 in each cell image as a predetermined image processing (hereinafter, "" "Trophoblast cell determination image processing") and image processing for determining the internal cell mass formed in the scutellum stage embryo (hereinafter, referred to as "internal cell mass determination image processing") are executed. Then, the quality of the trophoblast cells and the inner cell mass in the fertilized egg E imaged in the cell image is detected.

In addition, the fourth scoring processing unit 386 performs scoring based on the quality of the trophoblast cells and the inner cell mass, and scores the fourth stage using a plurality of cell images such as the average or the sum of each cell image. decide.

For example, in the case of the cell image of the fertilized egg E in the state stage of the fourth stage as shown in FIG. 14 (A), the second scoring processing unit 384 is based on the preconditions as in the other stages. , Edge detection processing was performed to detect embryos (images [4-1] and (B) in FIG. 14), and the uniformity of trophoblast cell thickness was evaluated ([4-] in FIG. 14). 2]), obtain the evaluation result of the trophoblast cell.

That is, the second scoring processing unit 384 executes edge detection processing to detect the edge of the embryo part, and based on the degree of agreement with the ideal texture created in advance, or the edge of the embryo part. While identifying the part of the trophoblast cell from the brightness value adjacent to the inside, the thickness is detected, the thickness is compared with the thickness range (optimal range) determined by the preconditions, and the nutrition is concerned. Evaluate trophoblasts.

Specifically, when the ideal texture created in advance is used, the second scoring processing unit 384 sets a filter of the luminance value created according to good, acceptable, and defective, and the detected embryo portion. The histogram of the brightness value of each pixel is compared with each other to calculate the degree of matching with the ideal texture, and the evaluation of the filter having the highest degree of matching is determined as the evaluation of the cell image.

Further, when the second scoring processing unit 384 evaluates from the brightness value adjacent to the inside of the edge of the embryo part, the average value of the total thickness in the trophoblast cells is within the optimum range, or , It is determined whether there is a portion thicker than the optimum range, and for example, the concave point detection process in the third phase is executed to detect the concave points in the trophoblast cells, and whether or not the number is within the predetermined range. The thickness of the trophoblast cells is evaluated on a three-point scale of good, acceptable, and defective.

In particular, when the fourth scoring processing unit 386 evaluates from the brightness value adjacent to the inside of the edge, the range of the brightness value regarded as the trophoblast cell (that is, corresponding to the thickness of the trophoblast cell) is optimal. If it is uniform within the range of, it is judged as "good", if it is not within the optimum range but within the predetermined range, it is judged as "OK", and in other cases, it is judged as "bad". To do.

The fourth scoring processing unit 386 may use other image processing as long as it can detect that the thickness of the trophoblast cells is uniform within a predetermined range.

On the other hand, when the embryo is detected by the edge detection process ([4-1] in FIG. 14), the fourth scoring processing unit 386 evaluates the clarity (concentration) of the inner cell mass (clarity in FIG. 14). Evaluation [4-3]), the evaluation result of the inner cell mass is obtained.

That is, when the fourth scoring processing unit 386 executes the edge detection process to detect the edge of the embryo portion, as shown in FIG. 15, a predetermined condition is satisfied in the brightness value of each pixel existing inside the edge. The included pixels are detected as the pixels constituting the inner cell mass, and the brightness values of the detected pixels are further histogramd, and the hiss and grams in three stages of good, acceptable, and defective are converted into data as preconditions in advance. The inner cell mass is evaluated by comparing the data with the histogramd data.

It should be noted that, as the three-stage hiss and gram data of good, acceptable, and defective that have been converted into data as preconditions in advance, those generated by machine learning such as a support vector machine are used.

Further, for example, in the present embodiment, when the fourth scoring processing unit 386 is within the range where the hiss and gram data of the inner cell mass are the same as or assumed to be the same as the data determined to be “good”. , If the hiss and gram data of the inner cell mass are the same as or assumed to be the same as the data judged as "OK", it is determined as "OK" and the inner cell mass is determined. If the hiss and gram data of the above are the same as or within the range assumed to be the same as the data judged to be "bad", it is judged to be "bad".

On the other hand, when the fourth scoring processing unit 386 detects the trophoblast cells and the inner cell mass, it determines the score of the fourth stage based on each evaluation of the detected trophoblast cells and the inner cell mass. In particular, the fourth scoring processing unit 386 determines the score according to the combination of each of the three-stage evaluation of the trophoblast cell and the inner cell mass.

For example, the fourth scoring processing unit 386 determines the score by setting one "good" as "5 points", one "OK" as "3 points", and one "bad" as "0 points".

[3] Operation of Cell Quality Evaluation System Next, the operation of the cell quality evaluation system 1 of the present embodiment will be described with reference to FIGS. 16 to 19. 16 to 19 are flowcharts showing scoring processes executed in the first to fourth periods, respectively.

In this operation, it is assumed that the data shown in FIGS. 5 and 6 have already been recorded in the state stage management DB 302 and the scoring information DB 303, and the information is not registered in the cultured cell management DB 301. It is assumed that

Further, in this operation, a plurality of cell images obtained by capturing the fertilized egg E three times in each state stage are used, and the average value thereof is used as the score of each state stage.

[3.1] Operation start and first stage scoring process First, after the user collects sperms and eggs from the patient and completes the operation for insemination, the cultured cell management unit 382 sets the operation unit 390. When an input operation to register the fertilized egg E fertilization date and time, the cultured petri dish ID and the fertilized egg ID is detected (step Sa101), the input information is recorded in the ROM / RAM 320 and stored in the cultured cell management DB 301. Record the information (step Sa102).

Next, the imaging management unit 381 reads the imaging timing from the state stage management DB 302 and records it in the ROM / RAM 320 (step Sa103), and outputs a control signal to the timer 360 to start measuring the elapsed time (step Sa104). ..

Next, the imaging management unit 381 compares the elapsed time being measured by the timer 360 with the imaging timing recorded in the ROM / RAM 320, and determines whether or not the first imaging timing has arrived (step Sa105). ..

At this time, when the imaging management unit 381 detects the imaging timing, it outputs a control signal to the camera 10, causes the fertilized egg E to be imaged, and acquires image data corresponding to the cell image from the camera 10 (step Sa106). The image pickup management unit 381 records the acquired image data in the ROM / RAM 320.

Next, the imaging management unit 381 determines whether or not all the imaging in the first phase is completed (step Sa107). At this time, the imaging management unit 381 shifts to the process of step Sa105 when all the imaging of the first period is not completed, and when it is determined that all the imaging of the first period is completed, step Sa108. Move to the processing of.

By repeating the process of steps Sa105 to 107, the image pickup management unit 381 images the fertilized egg E at each elapsed timing of 6 hours, 7 hours, and 8 hours, and sequentially corresponds to the ROM / RAM 320. Record the image data to be used.

Next, when it is determined that all the imaging of the first period is completed, the first scoring processing unit 383 reads out the scoring reference information corresponding to the first period from the scoring information DB 303 (step Sa108).

Next, the first scoring processing unit 383 executes the first scoring process on the image data recorded in the ROM / RAM 320 according to the read scoring reference information and the algorithm, and sets the score of the first period. The score is determined and the score of the determined first stage is recorded in the corresponding region of the cultured cell management DB 301 (step Sa109).

Specifically, the first scoring processing unit 383 detects the number of polar bodies contained in each cell image based on the algorithm of the first stage, and based on the number of polar bodies and scoring reference information in each cell image, While determining the score of each cell image, the score of the first phase is determined. In addition, the first scoring processing unit 383 sets the score value according to the growth rate of the fertilized egg E, that is, according to the time reception when two polar bodies are generated, for example, "+ 20%" and "0%". , "-20%" to change the score of the first period.

Next, the first scoring processing unit 383 determines whether or not the calculated score of the first period exceeds the second threshold value (step Sa110). At this time, when the first scoring processing unit 383 determines that the score of the first period is lower than the second threshold value, it determines that the fertilized egg E is not suitable for transplantation, and the fertilized egg E is not suitable for transplantation. End the process without scoring. Further, when the first scoring processing unit 383 determines that the score of the first period exceeds the second threshold value, the first scoring processing unit 383 shifts to the processing of step Sb101 in order to execute the scoring processing of the second period. ..

In this operation, since the score of the first stage greatly affects the quality of the fertilized egg E, the second threshold value is set to, for example, "10 points or more". In addition, when the score of the first stage, which has a large effect on the quality of the fertilized egg E by executing such a process, falls below the second threshold value, the fertilized egg E cannot be used for transplantation. By determining, it is possible to omit the processing after the second period and reduce the calculation load of the scoring processing.

[3.2] Second Phase Scoring Process First, the imaging management unit 381 reads the imaging timing of the second phase from the state stage management DB 302 and records it in the ROM / RAM 320 (step Sb101).

Next, the imaging management unit 381 compares the elapsed time being measured by the timer 360 with the imaging timing recorded in the ROM / RAM 320, and determines whether or not the second imaging timing has arrived (step Sb102). ).

At this time, when the imaging management unit 381 detects the imaging timing, it outputs a control signal to the camera 10, causes the fertilized egg E to be imaged, and acquires image data corresponding to the cell image from the camera 10 (step Sb103). The image pickup management unit 381 records the acquired image data in the ROM / RAM 320.

Next, the imaging management unit 381 determines whether or not all the imaging in the second phase is completed (step Sb104). At this time, the imaging management unit 381 shifts to the process of step Sb102 when all the imaging in the second phase is not completed, and when it is determined that all the imaging in the second phase is completed, the imaging management unit 381 shifts to step Sb105. Move to the processing of.

By repeating the processes of steps Sb102 to 104, the image pickup management unit 381 images the fertilized egg E at each elapsed timing of 19 hours, 20 hours, and 21 hours, and sequentially puts the image of the fertilized egg E into the ROM / RAM 320. Record the corresponding image data.

Next, when it is determined that all the imaging of the second period is completed, the second scoring processing unit 384 reads out the scoring reference information corresponding to the second period from the scoring information DB 303 (step Sb105).

Next, the second scoring processing unit 384 executes the second scoring process on the image data recorded in the ROM / RAM 320 according to the read scoring reference information and the algorithm, and sets the score of the second term. It is determined, and the determined second stage score is recorded in the corresponding region of the cultured cell management DB 301 (step Sb106).

Specifically, the second scoring processing unit 384 detects the number of polar bodies and pronuclei contained in each cell image based on the algorithm of the second phase, and the number of polar bodies and pronuclei in each cell image. And, based on the scoring standard information, the score of the second phase is determined while determining the score of each cell image. In addition, the second scoring processing unit 384 sets the score value according to the growth rate of the fertilized egg E, that is, according to the time when two polar bodies and pronuclei are generated, for example, "+ 20%", "+ 20%", " The score of the second period is changed as "0%" and "-20%".

Next, the second scoring processing unit 384 determines whether or not the calculated score of the second period exceeds the third threshold value (step Sb107). At this time, when the second scoring processing unit 384 determines that the score of the second period is lower than the third threshold value, it determines that the fertilized egg E is not suitable for transplantation, and the fertilized egg E is not suitable for transplantation. End the process without scoring. Further, when the second scoring processing unit 384 determines that the score of the second period exceeds the third threshold value, the second scoring processing unit 384 shifts to the processing of step Sc101 in order to execute the scoring processing of the third period. ..

In this operation, the score in the second period greatly affects the quality of the fertilized egg E as in the first period, so that the third threshold value is set to, for example, "10 points or more". In addition, when the score of the second stage, which has a large effect on the quality of the fertilized egg E by executing such a process, falls below the third threshold value, the fertilized egg E cannot be used for transplantation. By determining, it is possible to omit the processing after the third period and reduce the calculation load of the scoring processing.

[3.3] Third Phase Scoring Process First, the imaging management unit 381 reads the imaging timing of the third phase from the state stage management DB 302 and records it in the ROM / RAM 320 (step Sc101).

Next, the imaging management unit 381 compares the elapsed time being measured by the timer 360 with the imaging timing recorded in the ROM / RAM 320, and determines whether or not the third imaging timing has arrived (step Sc102). ).

At this time, when the imaging management unit 381 detects the imaging timing, it outputs a control signal to the camera 10, causes the fertilized egg E to be imaged, and acquires image data corresponding to the cell image from the camera 10 (step Sc103). The image pickup management unit 381 records the acquired image data in the ROM / RAM 320.

Next, the imaging management unit 381 determines whether or not all the imaging in the third phase is completed (step Sc104). At this time, the imaging management unit 381 shifts to the process of step Sc102 when all the imaging in the third period is not completed, and when it is determined that all the imaging in the third period is completed, step Sc105 Move to the processing of.

By repeating the processes of steps Sc102 to 104, the imaging management unit 381 captures an image of the fertilized egg E at each elapsed timing in each of the split state stages of the first half, the middle and the second half, and the ROM / RAM 320 has an image of the fertilized egg E. The corresponding image data is sequentially recorded.

Next, when it is determined that all the imaging of the third period is completed, the third scoring processing unit 385 reads out the scoring reference information corresponding to the third period from the scoring information DB 303 (step Sc105).

Next, the third scoring processing unit 385 executes the third period scoring process on the image data recorded in the ROM / RAM 320 according to the read scoring reference information and the algorithm, and sets the score of the third period. The score is determined, and the determined third stage score is recorded in the corresponding region of the cultured cell management DB 301 (step Sc106).

Specifically, the third scoring processing unit 385 detects the number of blastomeres contained in each cell image for each divided state stage, that is, in each of the prophase, metaphase, and anaphase, based on the algorithm of the third phase. , While determining the score of each cell image based on the number of blastomeres and the scoring standard information, each score of the prophase, metaphase, and anaphase is determined. In addition, the third scoring processing unit 385 determines the specified number of blastomeres generated in each divided state stage (2 in the early stage, 4 in the middle stage, and 8 in the late stage) according to the growth rate of the fertilized egg E. Depending on the time to reach (pieces), the score value is set to, for example, "+ 20%", "0%", and "-20%", and the score of the third period is changed. Then, the third scoring processing unit 385 averages the scores of each divided state stage to determine the overall score of the third period.

Next, the third scoring processing unit 385 determines whether or not the calculated score of the third period exceeds the fourth threshold value (step Sc107). At this time, when the third scoring processing unit 385 determines that the score of the third period is lower than the fourth threshold value, it determines that the fertilized egg E is not suitable for transplantation, and after the third period. End the process without scoring. Further, when the third scoring processing unit 385 determines that the score of the third period exceeds the fourth threshold value, the third scoring processing unit 385 shifts to the processing of step Sd101 in order to execute the scoring processing of the fourth period. ..

In this operation, by executing such a process, when the score of the third stage falls below the fourth threshold value, it is determined that the fertilized egg E cannot be used for transplantation. It is possible to omit the processing after the 4th period and reduce the calculation load of the scoring processing.

[3.4] Scoring process of the 4th period and comprehensive evaluation of fertilized egg First, the imaging management unit 381 reads the imaging timing of the 4th period from the state stage management DB 302 and records it in the ROM / RAM 320 (step Sd101). ..

Next, the imaging management unit 381 compares the elapsed time being measured by the timer 360 with the imaging timing recorded in the ROM / RAM 320, and determines whether or not the fourth imaging timing has arrived (step Sd102). ).

At this time, when the imaging management unit 381 detects the imaging timing, it outputs a control signal to the camera 10, causes the fertilized egg E to be imaged, and acquires image data corresponding to the cell image from the camera 10 (step Sd103). The image pickup management unit 381 records the acquired image data in the ROM / RAM 320.

Next, the imaging management unit 381 determines whether or not all the imaging in the fourth period is completed (step Sd104). At this time, the imaging management unit 381 shifts to the process of step Sd102 when all the imaging in the fourth period is not completed, and when it is determined that all the imaging in the fourth period is completed, step Sd105 Move to the processing of.

By repeating the processes of steps Sd102 to 104, the image pickup management unit 381 takes an image of the fertilized egg E at each elapsed timing, and sequentially records the corresponding image data in the ROM / RAM 320.

Next, when it is determined that all the imaging of the fourth period is completed, the fourth scoring processing unit 386 reads out the scoring reference information corresponding to the fourth period from the scoring information DB 303 (step Sd105).

Next, the 4th scoring processing unit 386 executes the 4th period scoring process on the image data recorded in the ROM / RAM 320 according to the read scoring reference information and the algorithm, and sets the score of the 4th period. It is determined, and the determined fourth stage score is recorded in the corresponding region of the cultured cell management DB 301 (step Sd106).

Specifically, the fourth scoring processing unit 386 detects the trophoblast cells and the inner cell mass contained in each cell image based on the algorithm of the fourth phase, and the trophoblast cells and the inner cell mass in each cell image. And, based on the scoring standard information, the score of the 4th period is determined while determining the score of each cell image.

In particular, the fourth scoring processing unit 386 calculates the thickness and uniformity of the trophoblast cells in the detected trophoblast cells and inner cell mass in the cell image based on the algorithm of the fourth phase, and also calculates the trophoblast cells. The state of is compared with the scoring standard information (ideal state and poor state) of the 4th stage, the trophoblast cells are evaluated in 3 stages of good, good and bad, and the size and clarity of the inner cell mass. The score (diaphragm based on the brightness value) is compared with the scoring reference information (diaphragm of ideal state and bad state) of the 4th period, and the trophoblast cells are evaluated in 3 stages of good, good and bad.

Further, the fourth scoring processing unit 386 sets the score value to, for example, "+ 20%", "0%", according to the quality evaluation (specifically) of the trophoblast cell and the inner cell mass at each timing. The score of the 4th period is changed as "-20%".

Next, the fertilized egg evaluation unit 387 calculates the sum of the scores at each state stage of the fertilized egg to be evaluated, calculates the total score (final evaluation result) (step Sd110), and records it in the cultured cell management DB 301. , The total score is displayed on the display unit 330 in conjunction with the display control unit 340 (step Sd111), and this operation is terminated.

In addition, in the process of step Sd111, when there is another fertilized egg E in the same patient, the fertilized egg evaluation unit 387 compares the total score of the other fertilized egg E recorded in the cultured cell management DB 301 and comprehensively compares them. The fertilized eggs may be sorted and displayed in descending order of score, or when the total score is lower than the first threshold value as compared with the first threshold value, the fertilized egg is evaluated as unusable for transplantation, and the first threshold value is evaluated. If it is equal to or higher than the threshold value of, it may be evaluated as transplantable.

As described above, the cell quality evaluation system 1 of the present embodiment can automatically evaluate the quality of the target fertilized egg E by imaging the fertilized egg E with the passage of time. Therefore, it is possible to reduce the workload of the user (for example, a doctor or an operator) at the time of quality evaluation of the fertilized egg E, and by scoring the cells that are in different states for each state stage to each state stage, the cells Since it is possible to evaluate the superiority or inferiority of the quality of the cells, it is possible to accurately evaluate the cells in which the growth process is important in the evaluation of the quality.

That is, since the internal structure of the cell changes according to the state stage and the quality determination standard also changes according to the state stage, the cell quality evaluation system 1 of the present embodiment has different scoring standard information for each state stage. And while applying the algorithm, scoring can be performed, and appropriate scoring can be realized according to the state stage.

Therefore, the cell quality evaluation system 1 of the present embodiment can automatically and highly accurately evaluate the quality of the cell to be evaluated.

In addition, the cell quality evaluation system 1 of the present embodiment accurately identifies cells having a high growth rate by performing scoring based on the images of cells taken at different timings in each state stage. The quality of cells can be evaluated with high accuracy.

Further, the cell quality evaluation system 1 of the present embodiment can evaluate, for example, a cell whose score at any state stage is lower than a predetermined threshold value as unusable for transplantation, and thus the later state. It is possible to omit scoring at the stage and reduce the processing load at the time of evaluation.

[4] Modification example [4.1] Modification example 1
In the above embodiment, the server device 30 is provided with DB 301 to DB 303, but these DB 301 to DB 303 may be managed and operated by different computer systems.

[4.2] Modification 2
In the above embodiment, the case where the sum of the scores in each state stage is used as the total score has been described as an example, but in step Sd110, the average or the root mean square of the scores in all state stages is calculated and used as the total score. Scores may be used.

In addition, weighting is performed in each state stage (including the divided state stage) when calculating the total score, and for example, the weighting of the first and second stages, which greatly affects the quality of the fertilized egg E, is increased. May be good. Even in this case, the total score can be calculated by using any of the sum, the average, and the root mean square.

Further, for example, the quality of the fertilized egg E is highly dependent on the quality of the first to third stages (excluding the latter stage) and the fourth stage, so that a plurality of sums of scores corresponding to these specific state stages are added. , Average and root mean square, etc., and determine whether or not the conditions are met, such as comparing with a predetermined threshold. If the conditions are not met, the fertilized egg E is used for transplantation. It may be evaluated as impossible.

[4.3] Modification 3
The fertilized egg evaluation unit 387 of the above embodiment sorts in the order of the best overall score, presents it to the user, and uses it as a reference when the user selects the fertilized egg. A fertilized egg E having an extremely low score of "0 point" or less may be evaluated as unsuitable for transplantation and may be displayed on the display unit 330 to the effect that it cannot be used.

In this case, for example, when the list of fertilized eggs E is sorted and displayed, the fertilized egg E is displayed to the user by displaying a flag indicating that the fertilized eggs are unusable in association with the unusable fertilized eggs. It can be shown that it is unavailable.

[4.4] Modification 4
Further, the imaging timing recorded in the state stage management DB 302 may be observed in advance according to the culture environment (medium type, temperature, acidity, etc.) to determine the optimum imaging timing.

Even in this case, it is possible to identify whether the fertilized egg grows fast or slow by performing imaging one hour before and after the imaging timing.

[4.5] Modification 5
In the above embodiment, when the score corresponding to each state stage falls below the second threshold value to the fourth threshold value, the scoring process in the later state stage is not automatically performed. When the threshold value falls below the threshold value, the display unit 330 may be displayed to that effect so that the user can determine the execution of the subsequent processing.

[4.6] Modification 6
In the above embodiment, the score is calculated using three cell images for each state stage, but a single cell image may be used, or four or more cell images may be used.

[4.7] Modification 7
Although the cell quality evaluation system of the above embodiment has been described when applied to a fertilized egg, it can also be applied to other cells whose internal state changes due to growth.

E ... Fertilized egg 10 ... Camera 30 ... Server device 300 ... Recording unit 301 ... Cultured cell management DB
302 ... Status stage management DB
303 ... Scoring information DB
310 ... Interface unit 320 ... ROM / RAM
330 ... Display unit 340 ... Display control unit 360 ... Timer 370 ... Server management control unit 380 ... Data processing unit 381 ... Imaging management unit 382 ... Cultured cell management unit 383 ... 1st scoring processing unit 384 ... 2nd scoring processing unit 385 ... 3rd scoring processing unit 386 ... 4th scoring processing unit 387 ... Fertilized egg evaluation unit 390 ... Operation unit

Claims (13)

With the passage of time from the insemination of sperm cells to the egg cells, the state stage indicating the internal state changes stepwise .
The transition of the state stage is
(1) At least the first state stage in which the plasma membrane, the zona pellucida, and the polar body occur.
(2) A second state stage in which a pronucleus is generated in the plasma membrane,
(3) A third state stage in which the pronucleus disappears and cleavage occurs sequentially inside the zona pellucida.
(4) A fourth state stage in which the zona pellucida and the plasma membrane disappear and the trophoblast cells and the inner cell mass are transformed into blastocysts.
In a cell quality evaluation system that evaluates the quality of cells that transition in the order of
By imaging the image of the cell and controlling the imaging means for generating the image data, the image of the cell is imaged at a predetermined timing from insemination, and each of the first to fourth state stages is performed. An imaging control means that generates the corresponding image data, and
The scoring reference information, which is a reference for evaluating the quality of cells in each state stage by a score based on the image of the cell corresponding to the generated image data, is the state stage identification information for identifying each state stage. A recording control means that controls the recording means to be recorded in association with
An acquisition means for acquiring image data output from the image pickup means, and
For each of the state stage, while a predetermined image processing on the acquired cell image, based on the relevant scoring criteria information, and scoring means for scoring the quality of the corresponding cell,
An evaluation means for evaluating the superiority or inferiority of the quality of the cells according to the scoring result indicating the scored result, and
A presentation means for presenting the evaluation result of the evaluation means to the user in a viewable manner,
A cell quality evaluation system characterized by comprising. In the cell quality evaluation system according to claim 1,
A cell quality evaluation system in which the scoring means scores the quality of a corresponding cell based on a plurality of cell images having different timings for each state stage. In the cell quality evaluation system according to claim 2,
The scoring means scores the quality of the corresponding cell for each state stage while weighting each cell image generated by being imaged at a different timing for each state stage according to the elapsed time. , Cell quality evaluation system. In the cell quality evaluation system according to claim 3,
A cell quality evaluation system using an average score obtained by averaging the scores of cell images at different timings with different elapsed times as the scores of the respective state stages. In the cell quality evaluation system according to claim 3,
A cell quality evaluation system using a representative score of 1 selected from the respective scores of cell images at each timing with different elapsed times as the score of each state stage. In the cell quality evaluation system according to any one of claims 1 to 5,
When the evaluation means determines whether or not the predetermined first condition is satisfied based on the sum of the scores scored for each state stage, and does not satisfy the first condition. In addition, a cell quality evaluation system that determines that the corresponding cell cannot be used as the superiority or inferiority of the quality of the cell. In the cell quality evaluation system according to any one of claims 1 to 6,
It is determined whether or not the evaluation means satisfies the second condition predetermined for each state stage for each score scored for each state stage, and the second condition is not satisfied. A cell quality evaluation system that determines that the corresponding cell is unusable. In the cell quality evaluation system according to claim 7,
When any of the scores in each of the state stages does not satisfy the second condition, the evaluation means performs scoring in the scoring means of the state stage that arrives after the state stage. A cell quality assessment system to discontinue. In the cell quality evaluation system according to any one of claims 1 to 8.
A cell quality evaluation system in which the evaluation means evaluates the superiority or inferiority of the quality of the corresponding cells while weighting the score of each scored state stage for each state stage. In the cell quality evaluation system according to any one of claims 1 to 9,
When the imaging means is imaged for each of a plurality of cells and a cell image corresponding to each of the state stages is generated.
The scoring means calculates a score corresponding to each state stage for each of the cells using the corresponding cell image.
The evaluation means evaluates the superiority or inferiority of the quality of the cells according to the scoring result for each cell, and ranks the values indicated by the scoring results of the cells in descending order.
A cell quality evaluation system in which the presentation means presents the evaluation results of each ranked cell so that the user can view them. With the passage of time from the insemination of sperm cells to the egg cells, the state stage indicating the internal state changes stepwise .
The transition of the state stage is
(1) At least the first state stage in which the plasma membrane, the zona pellucida, and the polar body occur.
(2) A second state stage in which a pronucleus is generated in the plasma membrane,
(3) A third state stage in which the pronucleus disappears and cleavage occurs sequentially inside the zona pellucida.
(4) A fourth state stage in which the zona pellucida and the plasma membrane disappear and the trophoblast cells and the inner cell mass are transformed into blastocysts.
In a program for evaluating the quality of cells transitioning in the order of
Computer,
By imaging the image of the cell and controlling the imaging means for generating the image data, the image of the cell is imaged at a predetermined timing from insemination, and each of the first to fourth state stages is performed. An imaging control means that generates the corresponding image data,
The scoring reference information, which is a reference for evaluating the quality of cells in each state stage by a score based on the image of the cell corresponding to the generated image data, is the state stage identification information for identifying each state stage. A recording control means that controls the recording means to be recorded in association with
An acquisition means for acquiring image data output from the imaging means,
A scoring means for scoring the quality of a corresponding cell based on the corresponding scoring reference information while performing a predetermined image processing on the acquired cell image for each state stage.
An evaluation means for evaluating the superiority or inferiority of the quality of the cells according to the scoring result indicating the scored result, and
A presentation means for presenting the evaluation result in the evaluation means so that the user can view it.
A program characterized by functioning as. With the passage of time from the insemination of sperm cells to the egg cells, the state stage indicating the internal state changes stepwise .
The transition of the state stage is
(1) At least the first state stage in which the plasma membrane, the zona pellucida, and the polar body occur.
(2) A second state stage in which a pronucleus is generated in the plasma membrane,
(3) A third state stage in which the pronucleus disappears and cleavage occurs sequentially inside the zona pellucida.
(4) A fourth state stage in which the zona pellucida and the plasma membrane disappear and the trophoblast cells and the inner cell mass are transformed into blastocysts.
In the cell quality evaluation method for evaluating the quality of cells transitioning in the order of
And imaging the cells corresponding to each of the first through fourth states stages to generate image data,
The scoring reference information, which is a reference for evaluating the quality of cells in each state stage by a score based on the image of the cell corresponding to the generated image data, is the state stage identification information for identifying each state stage. Control the recording means to be recorded in association with
For each state stage, the quality of the corresponding cell is scored based on the corresponding scoring reference information while performing a predetermined image processing on the generated cell image.
The superiority or inferiority of the quality of the cells is evaluated according to the scoring result indicating the scored result.
A cell quality evaluation method, characterized in that the superiority or inferiority of the evaluated cell quality is presented as an evaluation result so that the user can view it. With the passage of time from the insemination of sperm cells to the egg cells, the state stage indicating the internal state changes stepwise .
The transition of the state stage is
(1) At least the first state stage in which the plasma membrane, the zona pellucida, and the polar body occur.
(2) A second state stage in which a pronucleus is generated in the plasma membrane,
(3) A third state stage in which the pronucleus disappears and cleavage occurs sequentially inside the zona pellucida.
(4) A fourth state stage in which the zona pellucida and the plasma membrane disappear and the trophoblast cells and the inner cell mass are transformed into blastocysts.
The cells by imaging the transition of the order, and an imaging means for generating image data,
A server device that controls the imaging means and evaluates the quality of the cells using the cell images based on each state stage.
Have,
The server device
An image pickup control means for controlling the image pickup means and
The scoring reference information, which is a reference for evaluating the quality of cells in each state stage by a score based on the image of the cell corresponding to the generated image data, is the state stage identification information for identifying each state stage. A recording control means that controls the recording means to be recorded in association with
An acquisition means for acquiring image data output from the image pickup means, and
A scoring means for scoring the quality of the corresponding cell based on the corresponding scoring reference information while performing predetermined image processing on the acquired cell image for each state stage.
An evaluation means for evaluating the superiority or inferiority of the quality of the cells according to the scoring result indicating the scored result, and
A presentation means for presenting the evaluation result in the evaluation means so that the user can view it,
A cell quality evaluation system characterized by being equipped with.

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