JP2015130805A - Growth information management system, and growth information management program - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a growth information management system and a growth information management program which reduce a correspondence error of a microwell with cell growth information, when acquiring the expansion image of the microwell, in a cell culture container with two or more microwells.SOLUTION: A growth information management system of the invention comprises: memory measure for at least making a cell culture container with two or more microwells, an expansion sample image including one microwell, a discrimination information which discriminates the microwell, and cell growth information in the microwells correspond to each other, and store them; and a registration means for at least making the expansion sample image, the discrimination information, and the growth information correspond to each other, and register them in the memory measure based on information in which an image capturing order of the two or more microwells is associated with the discrimination information.

Description

  The present invention relates to a system and a program for managing growth information of cells in a cell culture container (cells that require individual management such as a fertilized egg).

  The in vitro fertilized egg (zygote) is produced by fertilizing sperm and ovum in a culture system, and the fertilized egg goes through the cleavage, morula, and blastocyst stages, and then emerges from the zona pellucida It is possible to culture up to the blastocyst stage. Assistive reproductive technology (ART) to transfer a fertilized egg from the cleavage to the blastocyst stage to the uterus to give birth to a baby is not limited to the livestock region. Established in infertility medicine.

  However, the success rate of pregnancy by in vitro fertilization is not necessarily high. For example, in humans, the success rate of pregnancy is still about 25 to 35%. One of the causes is that the probability of obtaining a high-quality fertilized egg suitable for transplantation into the uterus in culture is not high. A cultured fertilized egg is individually observed with a microscope to determine whether it is a high-quality fertilized egg suitable for transplantation into the uterus.

  In in vitro fertilization, a microdrop method is often used in which a culture solution is dropped in a container, and a fertilized egg is placed in the container and cultured in vitro. Conventionally, in this microdrop method, a cell culture container having a single flat bottom surface and a diameter of 30 to 60 mm is used as a cell culture container, and a drop of the culture solution is placed on the bottom surface of the cell culture container. A method has been used in which a plurality of cells are prepared and cells are cultured therein.

  If a drop is created in a normal cell culture container, the position of the fertilized egg changes due to cell movement of the fertilized egg itself or convection in the drop, and it becomes difficult to identify the fertilized egg that has been cultured and observed in it. There was a problem. Therefore, a means for controlling the position of a fertilized egg has been demanded.

  In order to make the culture effect of fertilized eggs more efficient, it is considered preferable to use the interaction (paracrine effect) between fertilized eggs. In order to control the position of the fertilized egg while using these effects, a microwell having the same size as the fertilized egg is formed on the bottom surface of the cell culture container, and the culture solution is covered so as to cover a plurality of microwells. A system is known in which a drop is added and a fertilized egg is placed in a microwell filled with a culture solution and cultured. Thereby, while controlling the position of a plurality of fertilized eggs and enabling individual observation, a plurality of fertilized eggs can be cultured in a small amount of culture solution, and the paracrine effect can be utilized.

  On the other hand, in order to manage the growth state of individual fertilized eggs, it is necessary to identify individual microwells and to recognize the growth state of fertilized eggs in individual microwells. Conventionally, when managing growth information for each cell, such as a fertilized egg, a person distinguishes individual cells at a culture location and links them with growth information, or the feature amount is obtained from an image of the cell when observed under a microscope. Extraction was performed to identify and manage individual cells.

  However, human error is unavoidable with the above management method. For example, there is a mistake when a person distinguishes between individual microwells. In addition, when registering feature values (cell diameter, etc.) detected by calculation as growth information, transcription errors occur when there is no subtle difference between the cell features, and identification of individual microwells and individual microwells There was a possibility that an error in the association with the growth information of the cells in the inside occurred.

International Publication No. 2011/004568 Pamphlet

  The present invention relates to a growth information management system and growth information for reducing errors in correspondence between microwells and cell growth information when acquiring an enlarged image of the microwell in a cell culture container having a plurality of microwells. The purpose is to provide a management program.

The present inventors have found that in the cell culture container having a plurality of microwells, the above problem can be solved by associating the imaging order of the plurality of microwells with identification information for identifying the microwells.
That is, the present invention includes the following inventions.

(1) a cell culture container having a plurality of microwells;
A storage means for storing an enlarged specimen image including one microwell, identification information for identifying the microwell, and growth information of cells in the microwell in association with each other;
A registration unit that registers the enlarged specimen image, the identification information, and the growth information in the storage unit in association with each other based on information in which the imaging order of the plurality of microwells is associated with the identification information;
A growth information management system.

(2) The growth information management system according to (1), further comprising imaging support means for performing a guide when imaging the enlarged specimen image according to the imaging order.

(3) It further comprises well arrangement determining means for recognizing arrangement information relating to the plurality of microwells from image information including all of the plurality of microwells and determining whether the arrangement information matches design information of the microwell. The growth information management system according to (1) or (2).

(4) The growth information management system according to any one of (1) to (3), further comprising changing means for changing a file name of the image file of the enlarged specimen image according to a predetermined naming rule.

(5) The cell culture container has a plurality of first identifiers attached in pairs for each of the microwells, and the enlarged specimen image includes the pair of the first identifiers and the microwells. ,
(1) to (4) further comprising determination means for determining whether the identification information associated with the imaging order matches the identification information identifying the microwell associated with the first identifier. The growth information management system described in any one.

(6) A program for causing an information processing apparatus including at least a calculation means and a storage means to execute growth information management processing for a cell culture container having a plurality of microwells,
In the calculation means,
Based on information in which the imaging order of the plurality of microwells and identification information for identifying the microwell are associated with each other, the enlarged specimen image including one microwell, the identification information, and the growth of cells in the microwell A program for executing a registration process for registering at least information in association with the storage means.

(7) In the calculation means,
The program according to (6), further causing an imaging support process for performing a guide when imaging the enlarged specimen image according to the imaging order.

(8) In the calculation means,
Recognizing arrangement information regarding the plurality of microwells from image information including all of the plurality of microwells, and further executing a well arrangement determination process for determining whether the arrangement information matches design information of the microwells. The program according to 6) or (7).

(9) In the calculation means,
The program according to any one of (6) to (8), further causing a change process to change the file name of the image file of the enlarged specimen image according to a predetermined naming rule.

(10) The cell culture container has a plurality of first identifiers attached in pairs for each of the microwells, and the enlarged specimen image includes the pair of the first identifiers and the microwells. ,
(6) causing the computing means to further execute a determination process for determining whether the identification information associated with the imaging order matches the identification information identifying the microwell associated with the first identifier; The program according to any one of (9) to (9).

(11) a first information processing apparatus including image acquisition means for acquiring an enlarged specimen image including one microwell from a cell culture container having a plurality of microwells;
A second information processing apparatus comprising storage means for storing the enlarged specimen image, identification information for identifying the microwell, and growth information of cells in the microwell in association with each other;
With
The first information processing device associates at least the enlarged specimen image with the identification information based on information in which the imaging order of the plurality of microwells and the identification information are associated with each other. Sent to the processing unit,
The second information processing apparatus calculates the growth information from the enlarged specimen image, and registers the enlarged specimen image, the identification information, and the growth information in the storage unit at least in association with each other. .

  According to the present invention, when acquiring an enlarged image of a microwell in a cell culture container having a plurality of microwells, it is possible to reduce errors in association between the microwell and cell growth information.

It is a block diagram of one Embodiment of the growth information management system of this invention. It is the schematic which shows the upper side figure of the 1st example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 1st example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 1st example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 1st example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 1st example of a cell culture container. It is a conceptual diagram of the enlarged image which image | photographed the pair of the microwell and the identifier with the microscope. It is the schematic which shows the upper side figure of the 2nd example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 2nd example of a cell culture container. It is the schematic which shows the enlarged top view of the cell accommodating part in the 3rd example of a cell culture container. It is the schematic which shows one Embodiment of the sample information database of this invention. It is the schematic which shows the 1st example of the well ID corresponding | compatible table of this invention. It is the schematic which shows the 2nd example of the well ID corresponding | compatible table of this invention. It is the schematic which shows the 3rd example of the well ID corresponding | compatible table of this invention. It is the schematic which shows one Embodiment of the 2nd identifier corresponding | compatible table of this invention. It is the schematic which shows one Embodiment of the well design information table of this invention. It is the schematic which shows one Embodiment of the growth information database of this invention. It is the schematic which shows one Embodiment of the determination profile table of this invention. It is a conceptual diagram explaining one Embodiment of the outline extraction process of this invention. It is a conceptual diagram explaining the processing content of one Embodiment of the 1st identifier recognition process part of this invention. It is a conceptual diagram explaining the processing content of one Embodiment of the 1st identifier recognition process part of this invention. It is a conceptual diagram explaining the processing content of one Embodiment of the feature-value calculation process part of this invention. It is a conceptual diagram explaining the processing content of one Embodiment of the feature-value calculation process part of this invention. It is a figure which shows a blastocyst simply. It is a figure explaining the morphological evaluation of an inner cell mass and trophectoderm. It is a figure explaining the morphological evaluation of a pronuclear stage embryo. It is a figure explaining switching of the profile according to elapsed time. It is a flowchart explaining the flow of a process of one Embodiment of the growth information management system of this invention. It is a flowchart explaining the flow of the feature-value calculation process of the growth information management system of this invention. It is a figure which shows another embodiment of a growth information database.

  Hereinafter, examples in which the present invention is applied to a human fertilized egg will be described. For example, when cultivating a human fertilized egg, the growth stage of the fertilized egg is usually determined while culturing, thereby determining whether the fertilized egg is of good quality suitable for transplantation into the uterus. . The growth information management system according to the present invention associates the imaging order of a plurality of microwells with identification information for identifying the microwells, thereby associating the microwells with the cell growth information, Realize management.

  FIG. 1 shows a configuration diagram of an embodiment of the growth information management system of the present invention. The growth information management system 1 includes a cell culture container 100, an image acquisition device 110, a second identifier reading device 120, and a growth information management device 130. Hereinafter, each component of the growth information management system 1 will be described.

  As shown in FIG. 1, the cell culture container 100 has a first identifier 101 and a second identifier 102. FIG. 2A shows a top view of a first example of the cell culture container 100, and FIG. 2B is an enlarged top view of the cell accommodating portion of FIG. 2A. The cell culture container 100 of the present embodiment has a bottom portion 103 and a side wall 104, and has a cell accommodating portion 106 in which a plurality of microwells 105 for accommodating cells are arranged. In this example, the shape of the bottom 103 is circular when viewed from above. The side wall 104 is formed so as to surround the outer edge of the bottom portion 103. The shape of the bottom 103 is not particularly limited, and may be a polygonal shape such as a triangle or a quadrangle, or a shape similar to a circle (including a substantially circular shape, an elliptical shape, and a substantially elliptical shape).

  The side opposite the bottom 103 is open, and the shape of the opening is preferably the same as the shape of the bottom 103. As shown in FIG. 2A, as an example, an opening having a circular shape and an opening width of preferably 30 to 60 mm, particularly about 35 mm is used. This is the same size as a conventional cell culture vessel used for cell culture, can be easily produced from a general-purpose cell culture vessel, and is easy to adapt to existing culture devices. Are preferred. Moreover, the cell culture container 100 may have a lid | cover similarly to a normal cell culture container.

  The microwell 105 forms a recess having a wall surface and an opening, and may be a recess provided directly as a recess in the bottom 103 of the cell culture vessel 100 or a recess formed by a member protruding from the bottom 103. In the example of FIG. 2A, the cell accommodating portion 106 is formed by forming a recess in the central portion of the bottom portion 103. The cell housing portion 106 has a V-shaped cross section.

  When the microwell 105 is photographed at a high magnification and this is repeated to photograph a plurality of pairs, the orientation of the cell culture container 100 at the time of photographing must always be constant. It is preferable that the cell culture container 100 has a configuration for specifying the direction of the cell culture container 100 from the viewpoint of always keeping the direction of the cell culture container 100 when photographing. For example, as shown in FIG. 2A, a bar pattern 109 extending in the lateral direction is attached to the bottom 103 in order to recognize the “downward direction” in the vertical direction on the enlarged specimen image. The bar pattern 109 is formed in a convex shape with respect to the bottom 103. For example, the width of the bar pattern is 0.5 to 2 mm. Further, the concave portion 109a is provided on the side wall 104, and the direction of the cell culture container 100 can be recognized by touching the side wall 104 of the cell culture container 100 with a finger.

  In addition, a circular first top and bottom mark 107 is attached to the bottom portion 103. The first top and bottom mark 107 is also attached to recognize the top and bottom of the cell culture container 100 (vertical direction on the enlarged specimen image). If the first top and bottom mark 107 is too small, it will be difficult to see, and if it is too large, it will interfere with the cell accommodating portion 106, and therefore preferably has a size (diameter) of 1 to 5 mm. In addition, the depth of the first top and bottom mark 107 is preferably 0.01 to 0 from the viewpoint of being visually observable and not wasting oil (oil for preventing the evaporation of the medium) that is added at the time of observation. .05 mm.

  The first top and bottom mark 107 is formed by surface processing. In FIG. 2A, the black portion of the first top and bottom mark 107 is rough-finished and looks somewhat opaque. On the other hand, the white portion of the first top and bottom mark 107 is formed flat and looks transparent. In this way, by adding a rough portion and a flat portion that seem to be a pattern when the circular first top and bottom mark 107 is seen, the probability that it becomes difficult to see due to light adjustment is reduced. For example, since the reflected light with respect to a specific direction becomes strong in a flat part, the flat one may be extremely easy to see depending on the light source. In addition, since the rough portion is scattered to some extent by any light source, it looks uniform regardless of the light source conditions, and thus it is easy to stably secure the difference in appearance from the portion other than the circular first top and bottom mark 107. There is. Note that the surface processing for the first top and bottom mark 107 may be performed on the entire circular shape in consideration of the usage environment of the user's cell culture container 100, or may be rough as shown in FIG. 2A. You may combine a surface processed part and a flat part.

  In the example of FIG. 2A, four first top and bottom marks 107 are attached to the bottom portion 103. The four first top and bottom marks 107 are arranged one by one at the four vertices of a square surrounding the cell storage unit 106. For example, the four first top marks 107 have an angle formed by a straight line obtained by translating the bar pattern 109 described above and a straight line passing through the center of gravity of the first top mark 107 and the center of the bottom portion 103 at 45 degrees, 135. It is attached at a position of 225 degrees, 225 degrees, and 315 degrees. The top and bottom adjustment of the cell culture container 100 is mainly performed with the bar pattern 109, but even in that case, the cell culture container 100 may be inclined several degrees. By arranging the first top and bottom marks 107 one by one at the four vertices of the square surrounding the cell storage unit 106, it becomes easy to recognize an inclination of several degrees.

  Further, the second top and bottom mark 108 may be attached to the bottom 103. In the example of FIG. 2A, two second top and bottom marks 108 are attached along the bar pattern 109 so as to sandwich the concave portion 109a of the side wall 104. Similarly to the first top and bottom marks 107, the second top and bottom marks 108 are subjected to surface processing. The second vertical mark 108 is formed with a smaller diameter than the first vertical mark 107. By recognizing both the first top mark 107 and the second top mark 108 by the reflection of light, it becomes easy to adjust the orientation of the cell culture container 100 without using the bar pattern 109. .

  As shown in FIG. 2B, the cell storage unit 106 has a circular shape in a top view. As for the diameter of the cell accommodating part 106, 2-5 mm is preferable. If the size of the cell storage unit 106 is small, there is little blank space other than the microwell 105, which may make operation difficult. On the other hand, if it is too large, a large amount of medium solution is required and the amount of medium solution that is wasted increases. In the cell accommodating part 106, it is preferable that the several microwell 105 is arrange | positioned at the square lattice form or the close-packed form. By arranging in a square lattice shape or a close-packed shape, it becomes easy to specify the position of each microwell 105 in the bottom 103 of the cell culture vessel 100. In the example of FIG. 2B, the combination of the first identifier 101 makes it easier to specify the position of each microwell 105 and is easy to apply to the automation process.

  The arrangement of the plurality of microwells 105 may be an arrangement in which a part of the arrangement is omitted from a square lattice or a close-packed arrangement. For example, a case where eight or more microwells are arranged at equal pitches on the sides and vertices of the parallelogram to form a cell accommodation unit. Parallelograms include squares, rectangles, rhombuses and other parallelograms. As shown in FIG. 2B, in the cell culture container 100 of this example, eight microwells are arranged one by one at the four vertices of the square and one at the midpoint of the four sides. . Thereby, a cell, such as a fertilized egg, can be arranged one by one in one microwell 105, and a plurality of cells can be cultured.

The figure formed by the outer edge of the opening of the microwell 105 is not particularly limited, and is a polygonal shape such as a triangle and a quadrangle, a circle (including a circle, a substantially circle, an ellipse, and a substantially ellipse), or U Although the shape of a character etc. may be sufficient, Preferably it is circular. Note that the microwell 105 may have a shape in which a stepped concave portion is formed toward the center of the microwell 105. This is because cells and the like easily flow to the center position. As shown in FIG. 2B, in this example, the figure formed by the outer edge of the opening of the microwell 105 is circular. Area of the opening of each microwell 105 in top view of the cell culture vessel 100 is preferably 3 mm ² or less, more preferably 1 mm ² or less, still more preferably 0.5 mm ² or less, preferably 0.03 mm ² or more It is. In other words, the area of the opening of each microwell 105 in the top view is the area of the figure formed by the outer edge of the opening of the microwell 105.

  When cultivating a fertilized egg using the cell culture vessel 100 of the present invention, it is desirable to culture to the blastocyst stage, so the diameter of the circular opening is larger than the maximum size of the cell at the blastocyst stage. It is desirable to be. When the outer edge of the opening of the microwell is circular, the opening width is smaller than the pitch between the microwells. Therefore, the opening width of the opening of the microwell (or the diameter when the outer edge of the opening of the microwell is circular) is preferably 0.1 mm or more, more preferably 0.15 mm or more, and further preferably 0.2 mm. It is above, Preferably it is less than 0.6 mm, More preferably, it is less than 0.4 mm. The opening width of the opening of the microwell can be defined as X + m (where X represents the maximum cell diameter). Here, m is preferably 0.01 mm or more, and more preferably 0.02 mm or more. The pitch between the microwells is preferably 1 mm or less, more preferably 0.8 mm or less, and still more preferably 0.6 mm or less.

  The depth of the microwell 105 refers to a depth measured vertically from the opening of the microwell to the deepest portion, and is preferably 0.05 to 0.5 mm. If the depth of the microwell is too shallow, the cells may move when the cell culture container is transported or when the cells divide, and the cells may come out of the microwell range. It is set so that it can be kept inside. For example, in order to retain the cells in the microwell, the depth is preferably 1/3 or more of the maximum cell diameter, and more preferably 1/2 or more. On the other hand, if the depth is too deep, it becomes difficult to introduce the culture solution or cells into the microwell. Therefore, the value is appropriately set so that the value is not too deep while the cells are held in the microwell. For example, the upper limit of the depth can be 3 times or less the opening width of the opening of the microwell. Furthermore, in order to facilitate the introduction of the culture solution, the depth is preferably not more than 1 times the opening width of the microwell, and particularly preferably not more than 1/2.

  In the vicinity of the plurality of microwells 105, the first identifier 101 is attached in pairs for each microwell 105, and the relative position of the first identifier 101 with respect to the paired microwell 105 is the first. It is characterized by being different for each pair of the identifier 101 and the microwell 105. Since the relative position of the first identifier 101 with respect to the microwell 105 is different for each microwell 105, only one pair of the microwell 105 and the first identifier 101 is observed, so that the microwell 105 has a plurality of microwells 105. The position can be specified. Since the first identifier 101 is attached in the vicinity of each microwell 105, when observing at a high magnification, it is not necessary to greatly shift the observation position from the microwell 105, and quick observation is possible. Furthermore, even when the cells are photographed at a high magnification, the first identifier 101 is photographed together with the microwell 105 in the magnified specimen image, so there is no need to manually add information to the magnified specimen image, Complicated work can be avoided, and the risk of an association error due to an operator's mistake can also be avoided.

The position of the first identifier 101 attached in a pair with each microwell 105 may be inside or outside the microwell 105, but is preferably arranged outside the microwell 105. This is because if it is provided inside the microwell 105, the observation of the fertilized egg may be hindered and the culture performance of the fertilized egg may be affected. Preferably, as shown in FIG. 2B, the first identifier 101 is attached to the outside of the plurality of arranged microwells 105 and in the vicinity of the microwells 105. The size of the first identifier 101 is preferably smaller than the size of the microwell 105. More specifically, the area of the first identifier in the top view of a cell culture vessel, 30000Myuemu ² or less, preferably 15000Myuemu ² or less, more preferably 8000Myuemu ² or less, preferably 100 [mu] m ² or more.

  In addition, the first identifier 101 is attached sufficiently close to the paired microwell 105 so that it is clear which microwell 105 is paired with. Accordingly, each first identifier 101 is preferably attached so that the distance from the paired microwells 105 is the smallest among all the microwells 105. The distance between the first identifier 101 and the microwell 105 is defined as the distance between the centroid of the graphic formed by the opening of the microwell 105 and the centroid of the graphic formed by the first identifier 101. Therefore, the distance between the first identifier 101 and the microwell 105 is preferably larger than ½ of the opening width of the microwell 105 and smaller than the pitch between the microwells 105. Specifically, the distance between the identifier and the microwell is preferably 500 μm or less, more preferably 400 μm or less, and even more preferably 300 μm or less.

  As shown in FIG. 2B, for example, the shape of the first identifier 101 is a dot shape. The shape of the graphic formed by the first identifier 101 is not particularly limited. Examples of figures include figures such as letters, numbers, polygons, arrows, lines (bars), dots, QR codes, barcodes, and combinations thereof. Since the first identifier 101, which is preferably smaller in size than the microwell 105, is attached in the vicinity of the microwell 105 suitable for individually containing cells such as fertilized eggs, the shape of the first identifier 101 is A simple shape that is easy to mold is preferred. This is because the cell culture container 100 is often manufactured by injection molding, so that it is difficult to form a very complicated shape with a minute size. Even if the shape of the first identifier 101 is simple, that is, even if the information of the first identifier 101 itself is small, the position of each microwell 105 is specified by adding information on the relative position to the microwell 105 can do. In addition, if the first identifier 101 has a complicated shape, the yield in manufacturing the cell culture container 100 may decrease. However, by using a simple shape, a decrease in yield can be avoided and the manufacturing cost can be reduced. Can do. For example, in the example of FIG. 2B, the first identifier 101 is formed in a convex shape, and the height is preferably 0.01 to 0.05 mm. This is because if the height of the first identifier 101 is too low, it is difficult to see, but if it is too high, it may interfere with the flow of the medium and the handling of the fertilized egg.

  In addition, as shown in FIG. 2B, character strings (alphabet characters) 111 are paired for each microwell 105 in the vicinity of the plurality of microwells 105. Since the character string 111 for the microwell 105 is different for each microwell 105, the position of the microwell 105 can be identified. The character string 111 is formed in a convex shape, and the height is preferably 0.01 to 0.05 mm. This is because if the height of the character string 111 is too low, it is difficult to see, but if it is too high, it may interfere with the flow of the medium and the handling of the fertilized egg.

  Moreover, the size of the character string 111 is preferably a size that fits within a 0.1 to 0.5 mm square. If the size of the character string 111 is too small, it will be difficult to see, but if it is too large, it may interfere with the flow of the medium and the handling of the fertilized egg. Further, if the character string 111 is formed large, the number of the microwells 105 is affected. The interval between the character strings 111 (for example, the interval between A and B) is preferably 0.3 to 0.7 mm. If the interval is small, it is difficult to distinguish the character string 111. If the interval is too large, it is difficult to recognize which microwell 105 corresponds to the character string. In addition, what is attached to the microwell 105 is not limited to characters such as alphabets, but includes figures, figures such as polygons, symbols, and the like.

  Furthermore, a symbol (heart) 112 is attached to the center position of the plurality of microwells 105. When the symbol 112 is present at the center, it is easy to recognize the center position surrounded by the plurality of microwells 105 when observed with the image acquisition device 110 at a low magnification. This facilitates alignment when observed at a high magnification.

  3A and 3B are diagrams illustrating the relationship between the dot-shaped first identifier 101 and the microwell 105 in an easy-to-understand manner. In these figures, for the sake of explanation, alphabetic character strings and central graphics are omitted. As shown in FIG. 3B, in this example, the angle of the first identifier 101 with respect to the paired microwell 105 is different. The angle α of the first identifier 101 with respect to the microwell 105 can be defined as follows. For example, in the example shown in FIG. 3A and FIG. 3B, the angle α is defined as a straight line parallel to the straight line X and the microwell 105 It can be defined as an angle formed by a straight line Y passing through the center of gravity and the center of gravity of the first identifier 101. And the position of the specific microwell 105 in the plurality of microwells 105 can be specified by arranging the angle α to be different for each pair of the microwell 105 and the first identifier 101.

  4A and 4B are other examples of the first identifier 101. FIG. In this example, eight microwells 105 are arranged one by one at four vertices of a square and one by one at the midpoints of the four sides to constitute the cell storage unit 106. One linear first identifier 101 is attached in the vicinity of each microwell 105.

  FIG. 4B represents a microscopic image of a pair of microwell 105 and first identifier 101 at positions A, E, and H in FIG. 4A, respectively. In this example, since the angle α and / or the length of the line (bar) of the first identifier 101 with respect to the microwell 105 is different for each pair, simply observing the pair of microwells 105 and the first identifier 101, The position of each microwell 105 can be specified. Further, since each first identifier 101 is linear in the same direction and attached to the right side with respect to the microwell 105, the microwell 105 photographed at a high magnification and the first identifier 101 are taken. Also in the pair of enlarged images, the orientation of the cell culture container at the time of imaging can be specified based on the position of the first identifier 101 and the direction of the line (bar). The “direction” here is a rotation angle and is different from the angle α. For example, when the dots are replaced with lines (bars) in the examples of FIGS. 3A and 3B, the angle at which the line rotates (rotates) with respect to the straight line X at the dot position, that is, the angle formed by the straight line X and the identifier line. Sure. Using this amount of orientation information, the orientation of the cell culture vessel can be specified.

  FIG. 5A shows a top view of a second example of the cell culture container 100, and FIG. 5B is an enlarged top view of the cell accommodating part of FIG. 5A. Similar to the first example, the cell culture container 100 of this example has a bottom part 103 and a side wall 104, and a plurality of microwells 105 for containing cells are arranged on the bottom part 103. 106. Also in this example, the shape of the bottom 103 is circular. In the example of FIG. 5A, three top and bottom marks 121 are attached, and the shape is “+”. The size of the top and bottom marks 121 is preferably a size that fits within a 1 to 5 mm square. This is because if the top and bottom marks 121 are too small, it will be difficult to see, and if the top and bottom marks 121 are too large, it will interfere with the cell storage unit 106.

  Moreover, the top and bottom marks 121 are formed in a convex shape, and the height is preferably 0.01 to 2 mm. This is because if the height of the top / bottom mark 121 is too low, it is difficult to see, and if it is too high, the work may be disturbed. The three top and bottom marks 121 are arranged so as to surround the cell storage unit 106, and the top and bottom marks of the cell culture container 100 corresponding to the “down direction” of the top and bottom (up and down direction on the enlarged specimen image) are attached. It has not been. Therefore, it is possible to keep the orientation of the cell culture container 100 constant by arranging the direction in which the top and bottom marks are not attached in front of the operator.

  As shown in FIG. 5B, 25 microwells 105 are arranged in a 5 × 5 square lattice in the cell storage unit 106. In this example, the first identifier 101 as in the first example is not attached to the plurality of microwells 105. On the other hand, alphabets A to E are attached to the left side of the square lattice-shaped microwell 105 so as to correspond to each row so that each row of the square-lattice-shaped microwell 105 can be recognized. Further, numbers 1 to 5 are attached to the upper side of the square lattice-shaped microwells 105 so as to correspond to the respective columns so that each row of the square lattice-shaped microwells 105 can be recognized. Even with such a configuration, the processing of the present invention described below can be applied. In addition, what is attached in order to recognize a row and a column is not limited to characters and numbers, such as an alphabet, A figure, a symbol, etc., such as a polygon, are mentioned.

  FIG. 6 is an enlarged top view of a third example of the cell storage unit. In the cell storage unit 106, 25 microwells 105 are arranged in a 5 × 5 square lattice. In this example, letters and numbers for recognizing rows and columns are not attached. Even with such a configuration, the processing of the present invention described below can be applied.

  Next, the second identifier 102 will be described. A second identifier 102 is attached to the cell culture container 100. Examples of the second identifier 102 include characters, numbers, figures such as polygons, QR codes, barcodes, IC tags, dot patterns (coded patterns), and combinations thereof. Here, the dot pattern (coded pattern) technique will be briefly described. For example, the dot pattern is printed on the surface of a dedicated paper for an electronic pen. The dot pattern is a combination of vertical and horizontal 6 × 6 = 36 dots arranged on a lattice with an interval of about 0.3 mm. The vertical and horizontal 6 × 6 dots are arranged so as to form a unique pattern no matter which portion of the dedicated paper is taken 6 × 6 dots. A dot pattern formed by these 36 dots holds position coordinates (for example, on which position the dot pattern is located on the dedicated paper) and a dot pattern address that is a unique identifier for each dedicated paper. . Each dot is associated with predetermined information by the shift direction from the reference position of the lattice. That is, according to this technique, the dot pattern can be made to correspond to the second identifier 102 by making the shift direction from the reference position of the lattice of each dot correspond to characters, numbers, and the like. The dot pattern can be photographed by reading means such as an electronic pen with a built-in camera. The dot pattern technique described here is also described in Japanese Patent Application Laid-Open Nos. 2004-252607 and 2004-054375, and the techniques described in these documents can be used. The second identifier 102 may be attached to the side wall 104 or the bottom 103 of the cell culture container 100. In addition, when the cell culture container 100 includes a lid, the second identifier 102 may be attached to the lid. The second identifier 102 is different for each cell culture container 100, and the cell culture container 100 can be specified by recognizing the second identifier 102.

  Next, other components of the growth information management system 1 will be described. The image acquisition device 110 captures an enlarged specimen image including cells in the microwell 105 of the cell culture container 100 and the first identifier 101, and is, for example, a microscope. As the image acquisition device 110, for example, a device equipped with a 1/2 inch CCD element, 4, 10, and 20 times objective lens is often used.

  The second identifier reading device 120 is a device that reads the second identifier 102 of the cell culture container 100. As the second identifier reading device 120, a device corresponding to the second identifier 102 may be employed, and examples thereof include a known character / graphic recognition device, a camera, a barcode scanner, and an IC tag reader.

  The growth information management device 130 is configured by an information processing device such as a personal computer or a workstation. This information processing apparatus includes a processor such as a central processing unit (CPU), a storage device 131 such as a memory and a hard disk, an input unit 132 such as a keyboard and a mouse, and an output unit 133 such as a display. I have. In FIG. 1, the growth information management device 130 is shown as one information processing device, but the present invention is not limited to this. The constituent elements of the growth information management apparatus 130 may be distributed to a plurality of information processing apparatuses on the network. Various tables and databases may be stored in other information processing apparatuses or servers on a network.

  Next, various information used in the growth information management system 1 of the present embodiment will be described. The storage device 131 stores a specimen information database 141, a well ID correspondence table 142, a second identifier correspondence table 143, a well design information table 144, a growth information database 145, and a determination profile table 146. These various tables and databases will be described using a “table” structure in the following description, but they may not necessarily be represented by a data structure using tables, but may be represented by other data structures. Therefore, in order to show that the data structure does not depend on the data, hereinafter, various tables and databases may be simply referred to as “information”.

  FIG. 7 is an example of the sample information database 141. The specimen information database 141 stores various types of information related to cells in the cell culture container (cells that require individual management such as fertilized eggs). The specimen information database 141 includes a patient ID 701, a treatment date and time 702, a cell culture container ID (container ID) 703, and a well ID 704 as configuration items. The patient ID 701 is a number that uniquely identifies a patient. By associating the patient ID 701 with the cell culture container ID 703 and the well ID 704, it can be applied to ID management such as in the case of human infertility treatment. The treatment date and time 702 is a string of numbers indicating the treatment date and time. Although mentioned later, when determining the growth stage of a fertilized egg, it may determine based on the elapsed time from the operation date. In such a case, it is preferable to manage reference information such as the treatment date and time in the specimen information database 141. The cell culture container ID 703 is a number that uniquely identifies the cell culture container 100. The well ID 704 is an ID for identifying the position of each microwell 105 in the cell culture container 100. By using the well ID 704, it is possible to manage information on a specimen of a cell (eg, a fertilized egg) arranged inside the microwell 105.

  FIG. 8A is a first example of the well ID correspondence table 142. FIG. 8A is an example of a table corresponding to the example shown in FIG. 2B. In the well ID correspondence table 142, since the imaging order and the well ID are associated as described below, the well ID, the enlarged specimen image, and the cell growth information are recorded in association with each other by using this. It becomes possible. The well ID correspondence table 142 includes a well ID 801, information 802 indicating a relative position between the first identifier 101 and the microwell 105 (here, an angle α of the first identifier 101 with respect to the microwell 105), and a position of the microwell 105. Information 803 indicating the image capturing order and the shooting order 804 at least as configuration items. Note that the information of the imaging order 804 may be registered in advance, or may be in a form in which the operator inputs the information every time a magnified specimen image is captured and is temporarily registered. Further, the well ID correspondence table 142 may store information such as XY coordinates indicating the position of the microwell 105 with reference to a certain predetermined position, in addition to the information 803 indicating the position of the microwell 105. This information can be used for functions such as automatic shooting according to the shooting order. The well ID correspondence table 142 may include design information of the first identifier 101 that is paired with each microwell 105. The design information of the first identifier 101 is information such as the shape, size, and position of the first identifier 101.

  In the example of FIG. 8A, the value of the angle α may be set with a predetermined width. This is because an error may occur in image processing in the growth information management apparatus 130 described later. 8A shows only information on the angle α, but when the first identifier 101 is a line (bar), information (angle α) indicating the relative position between the first identifier 101 and the microwell 105 and / or Alternatively, information (such as a line length and direction) included in the first identifier 101 itself may be stored. As described above, examples of the first identifier 101 include characters, figures, polygonal shapes, arrows, lines (bars), dots, QR codes, barcodes, and combinations thereof. Therefore, the well ID correspondence table 142 only needs to store information related to the first identifier 101 corresponding thereto.

  FIG. 8B is a second example of the well ID correspondence table 142. FIG. 8B is an example of a table corresponding to the example shown in FIG. 5B. Therefore, in this example, the well ID correspondence table 142 does not hold the information of the first identifier 101. The well ID correspondence table 142 in FIG. 8B includes a well ID 801, information 803 indicating the position of the microwell 105, and an imaging order 804 as configuration items. In addition, as shown in FIG. 5B, information 803 indicating the position of the microwell 105 is defined by a combination of alphabets and numbers.

  FIG. 8C is a third example of the well ID correspondence table 142. FIG. 8C is an example of a table corresponding to the example shown in FIG. The well ID correspondence table 142 in FIG. 8C includes a well ID 801, information 803 indicating the position of the microwell 105, and an imaging order 804 as configuration items. In FIG. 6, since no character string or the like is attached around the microwell 105, in this example, the information 803 indicating the position of the microwell 105 is defined by a position in a 5 × 5 square lattice. With such a configuration, even when a character string or the like is not attached around the microwell 105, it is possible to associate with the well ID according to the imaging order. Further, it becomes possible to guide the acquisition position of the enlarged specimen image (the position of the microwell 105) according to the imaging order.

  As described above, a plurality of examples of the well ID correspondence table 142 have been described. However, each of these tables may be associated with information on the type of cell culture container described later. In medical settings, different types of cell culture vessels may be used. Therefore, it is preferable to hold the well ID correspondence table 142 according to the type of each cell culture container as described above. In the process in the growth information management device 130, after the type of the cell culture container is determined, the corresponding well ID correspondence table 142 is selected.

  FIG. 9 is an example of the second identifier correspondence table 143. The second identifier correspondence table 143 includes at least a cell culture container ID 901, second identifier information 902, and a container type 903 as configuration items. Using this table, each cell culture container 100 can be identified from the second identifier 102 attached to the cell culture container 100. The container type 903 is information indicating the type of cell culture container. As described above, the arrangement of the microwells 105, the presence or absence of the first identifier 101, and the like may change for each cell culture container. Accordingly, by having information on the container type 903, design information (information such as the alignment and number of microwells) corresponding to the container type 903 can be selected.

  FIG. 10 is an example of the well design information table 144. The well design information table 144 includes a container type 1001, alignment (arrangement) information 1002 of the microwell 105, information 1003 of the number of microwells 105, and a character string (alphabet in FIG. Information 1004 indicating whether or not there are alphabets and numbers in FIG. 5B and information 1005 indicating whether or not there is the first identifier 101 are included as configuration items. In addition to these, the well design information table 144 has other designs such as, for example, the shape of the microwell 105, the dimension of the outer edge of the opening of the microwell 105, the area of the opening of the microwell 105, and the pitch between the microwells 105. Information may be included.

  FIG. 11 is an example of the growth information database 145. The growth information database 145 stores the enlarged specimen image acquired from the image acquisition device 110 and various types of information related to this image. The growth information database 145 includes a cell culture container ID 1101, a well ID 1102, and growth information 1103 as constituent items. Thus, in the growth information database 145, the growth information 1103 is associated with the cell culture container ID 1101 and the well ID 1102. Growth information 1103 includes image information 1104, information 1105 indicating whether or not a fertilized egg exists in the well in the image, information 1106 indicating the first feature amount of the cell in the well in the image, It includes information 1107 indicating the second feature amount of the cells in the well and date 1108 when the image information was acquired. Information 1106 indicating the first feature amount is information on the diameter of the blast ball, and information 1107 indicating the second feature amount is information on the number of blast balls. In this example, information about two feature amounts is managed for the image information 1104. However, the present invention is not limited to this example, and one or more feature amounts may be managed. Also, the feature amount is not limited to the example of FIG. 11, and various types of feature amount will be described later. Furthermore, as growth information, information indicating a growth stage (grade information of classification such as Veek, Gardner, etc.) may be stored together with the feature amount.

  Accordingly, by referring to the growth information database 145, it is possible to manage which microwell corresponding to the enlarged specimen image of each image information 1104, and information on the growth stage of the cells in the microwell at the same time. Can be managed. Further, by referring to both the growth information database 145 and the sample information database 141, each image information 1104 corresponds to which microwell's enlarged sample image and to which patient's time and date of treatment. Can manage. In the present embodiment, the growth information database 145 is created. However, an item in which image information and growth information can be registered in the sample information database 141, and an enlarged sample image and growth information are registered in the sample information database 141. But you can.

  FIG. 12 is an example of the determination profile table 146. The determination profile table 146 stores determination information for determining a cell growth stage. The determination profile table 146 includes an application time 1201, a grade 1202, and a condition 1203 as configuration items. Although mentioned later, when determining the growth stage of a fertilized egg, it may determine based on the elapsed time from the operation date. The application time 1201 indicates the elapsed time from the treatment date and time (culture start date and time). The condition 1203 stores information on the feature amount that should be calculated from the enlarged specimen image at the time of application 1201 and information on the condition that the feature amount should satisfy. Thereby, according to the elapsed time from the operation date and time, it is possible to switch the characteristic amount of the cell to be calculated and the determination condition for determining the growth state of the fertilized egg. For example, in the example of FIG. 12, when the elapsed time is 2 to 3 days, the profile of the first record considering the evaluation of Veek is selected. When the elapsed time is 4 to 5 days, the profile of the second record in consideration of Gardner's evaluation is selected. Details of Veek and Gardner will be described later. The grade information when the condition 1203 is satisfied is stored in the grade 1202. Using this grade 1202 information, it becomes possible to determine the growth stage of fertilized eggs (or the quality of fertilized eggs).

  Next, the growth information management device 130 will be described in detail. As illustrated in FIG. 1, the growth information management device 130 includes a first identifier recognition processing unit 151, a second identifier recognition processing unit 152, a well arrangement determination processing unit 153, a photographing support processing unit 154, and a feature amount calculation. A processing unit 155 and a database creation processing unit 156 are provided. The growth information management device 130 uses these processing units 151 to 156 to photograph the microwell 105 in the predetermined photographing order by the image acquisition device 110, analyzes the photographed enlarged specimen image, and stores them in the various tables described above. Based on this, the whole image information, the enlarged specimen image information, the well ID, and the growth information are associated with each other. This makes it possible to uniquely associate the growth information with the microwell without making a mistake.

  In this embodiment, you may implement | achieve each process part 151-156 of the growth information management apparatus 130 as a function of the program run on a computer. That is, the processing units 151 to 156 may be stored in the memory as program codes, and the processing units 151 to 156 may be realized by the CPU executing the program codes. The processing units 151 to 156 will be described below.

  The first identifier recognition processing unit 151 receives an enlarged specimen image including the cells in the microwell 105 of the cell culture container 100 and the first identifier 101 from the image acquisition device 110, and executes an outline extraction process on the enlarged specimen image. To do. A technique known to those skilled in the art can be applied as the contour extraction process. As an example of the contour line extraction process, there is a process of executing a binarization process for converting an enlarged specimen image into a two-tone image of white and black and extracting a contour line from the binarized image. . In general, since the object to be observed is transparent and it is difficult to add a contrast to the boundary, after performing a differential operation on the change of the pixel value, a process of converting to a two-tone image may be executed. . Moreover, as another outline extraction process, you may use the technique as described in Unexamined-Japanese-Patent No. 2011-192109. In this document, the outline of a fertilized egg is determined by creating a reference profile and a candidate profile, and calculating the degree of similarity indicating the similarity between the reference profile and the candidate profile. You may apply to determination of the outline of a line or a 1st identifier. The first identifier recognition processing unit 151 forms the contour of the figure formed by the outer edge of the opening of the microwell 105 (hereinafter referred to as “outer contour”) and the first identifier 101 by the contour extraction process. The contour line of the figure is extracted, and a contour line extraction image is output (FIG. 13).

  Further, interpolation processing and / or region definition processing may be executed after the contour line extraction processing in the first identifier recognition processing unit 151. Interpolation processing is processing that estimates and interpolates a portion where the contour line is interrupted by combining a pattern prepared in advance and the contour line extraction image. For example, if only the outline extraction process is performed, the outline of the microwell 105 and the outline of the figure formed by the first identifier 101 may be interrupted. Therefore, the pattern of the shape of the microwell 105 and the pattern of the shape of the first identifier 101 are registered in advance, and the respective contour lines are interpolated by superimposing the respective patterns on the contour line extraction image. The area definition process is a process for defining an area surrounded by an outline. For example, when the contour line on the contour line extracted image is discontinuous, a region surrounded by the contour line interpolated after the contour line interpolation process is defined. By performing this region definition process, it is possible to determine a region for pattern matching to be performed later and a region for calculating the size or area of the region surrounded by the outline.

  The first identifier recognition processing unit 151 performs pattern detection processing on the contour line extracted image. Here, the pattern detection processing is to detect which contour line is the outer peripheral contour line of the microwell 105 in the contour line extraction image and which contour line is the contour line of the first identifier 101. is there. As the pattern detection process, a technique known to those skilled in the art can be applied. As an example, there is a method in which a template having the shape of the microwell 105 and a template having the shape of the first identifier 101 are stored in advance in the growth information management device 130, and pattern matching is performed using each template. The first identifier recognition processing unit 151 detects a portion having a predetermined similarity or higher with respect to each template as the contour line of the microwell 105 or the contour line of the first identifier 101. Note that the template may be appropriately enlarged / reduced according to the enlargement magnification of the image acquisition device 110.

  The processing for detecting the outer contour of the microwell 105 and the contour of the first identifier 101 in the contour extraction image is not limited to this, and other methods may be used. For example, the detection may be performed using the design information of the microwell 105 in the well design information table 144 and the design information of the first identifier 101 in the well ID correspondence table 142. For example, by comparing information such as the size and area of the figure formed by the contour line on the contour line extracted image with the design information of the microwell 105 and the design information of the first identifier 101, the outer contour line of the microwell 105 The contour line of the first identifier 101 may be specified.

  The 1st identifier recognition process part 151 performs the recognition process of the 1st identifier 101 with respect to an outline extraction image. The first identifier recognition processing unit 151 calculates an angle α of the first identifier 101 with respect to the microwell (FIG. 14). When the first identifier 101 is a line (bar), the first identifier recognition processing unit 151 may calculate the angle α and / or the length of the line (bar). Thereafter, the first identifier recognition processing unit 151 refers to the well ID correspondence table 142 based on the calculated angle α, and outputs a well ID corresponding to the calculated angle α.

  In addition, the process of the 1st identifier recognition process part 151 is not limited to this, Another method may be sufficient. In the process of the first identifier recognition processing unit 151, it is only necessary that the microwell 105 and the first identifier 101 can be associated with the well ID based on the relative positional relationship between the microwell 105 and the first identifier 101. As shown in FIG. A plurality of templates including the above may be prepared and template matching may be performed. If each template and the well ID are associated with each other in the well ID correspondence table, the enlarged specimen image can be associated with the well ID from the result of template matching.

  The second identifier recognition processing unit 152 refers to the second identifier correspondence table 143 based on the information of the second identifier 102 read by the second identifier reader 120, and outputs the cell culture container ID corresponding to the information. In addition to this method, a method may be used in which the cell culture container ID is recognized in advance by another method, and the second identifier 102 is read by the second identifier reader 120 to check the recognition accuracy. In addition, the second identifier recognition processing unit 152 selects the corresponding information in the well design information table 144 in the subsequent processing, and selects the corresponding well ID correspondence table 142. The type 903 is output.

  The well arrangement determination processing unit 153 obtains arrangement information of the microwells 105 from all the microwells 105 and low magnification images (hereinafter, all image information) including character strings (numbers, alphabets, etc.) around the microwells 105. calculate. For the detection of the microwell 105, the one described as the processing of the first identifier recognition processing unit 151 described above may be used. For character strings around the microwell 105, a known character recognition process such as an OCR process may be used. The well arrangement determination processing unit 153 recognizes the arrangement of the microwells 105, the number of the microwells 105, the character strings around the microwells 105, and the like from all the image information.

  By referring to the well design information table 144, it is possible to acquire well design information of the container type corresponding to the second identifier 102. Therefore, the well arrangement determination processing unit 153 determines whether various information recognized from the entire image information matches the well design information in the well design information table 144. The determination is made, for example, according to items in the well design information table 144. For example, the determination here is whether the number of microwells 105 is the same, whether the microwells 105 are arranged in a square lattice, whether a character string is recognized around the microwells 105, or the like. Another determination may also be made. For example, since the taken magnification and the size of the microwell 105 are known in advance, it is possible to determine whether or not all of the microwells 105 are in a predetermined XY coordinate range on the acquired image. Thereby, it can be determined whether the container type recognized from the 2nd identifier 102 and the container type of the cell culture container actually image | photographed correspond.

  Further, the well arrangement determination processing unit 153 determines whether or not there is a cell in the microwell 105 from all image information. For example, when there are no cells in the microwell 105, the pixel values in the microwell 105 are uniformly distributed in the enlarged specimen image. The well arrangement determination processing unit 153 determines whether there is a change in the pixel value in the microwell 105 and determines whether there is a cell in the microwell 105. This determination process is an example, and other known methods may be used as long as the presence or absence of cells is determined. The determination result here can be used for confirming the number of cells before taking an individual enlarged specimen image, or can be registered as information 1105 in the growth information database 145.

  The imaging support processing unit 154 performs various processes for guiding the imaging sequence when imaging the individual enlarged specimen images of each microwell 105, a process for associating the enlarged specimen image with the well ID based on the imaging order, and Then, a process for determining whether or not an individual enlarged specimen image has been acquired correctly is performed. If the imaging order is registered in the well ID correspondence table 142 in advance, the imaging support processing unit 154 guides the imaging order when the enlarged specimen image is acquired after the container type is determined. In addition, when the shooting order is not determined in advance, the shooting support processing unit 154 may display a screen for causing the output unit 133 to input the shooting order, and acquire the shooting order from input information on the screen.

  As a guide method, when the operator operates the image acquisition apparatus 110 using the input unit 132 and adjusts the focus so that the microwell 105 and the first identifier 101 are included in the frame, the output unit 133 such as a display is used. There is a method for displaying the position information of the microwell 105 to be photographed. It is not limited to display on the display, and the shooting order may be guided by other methods such as voice. As for the contents of the guide, “Give me a picture of the microwell at position A first” or after shooting the microwell at position A, “Next, take a picture of the right microwell. Please guide me. " Other contents may be used as long as the position of the microwell 105 to be photographed can be recognized.

  If the image acquisition device 110 has a function of automatically acquiring an image by specifying the position (XY coordinate) of the microwell to be imaged and the imaging magnification, the imaging support processing unit 154 performs the imaging. According to the order, the position of the microwell to be imaged (XY coordinates) and the imaging magnification may be designated, and the enlarged sample image may be automatically acquired by the image acquisition device 110. In this case, information such as the XY coordinates of each microwell 105 may be held in the well ID correspondence table 142 or the like.

  The imaging support processing unit 154 associates the enlarged specimen image with the well ID by referring to the well ID correspondence table 142 in which the well ID and the imaging order are defined. For example, in the example of FIG. 8A, the first magnified specimen image is associated with the well ID “001”, and the second magnified specimen image is associated with the well ID “002”. By this association, the well ID, the enlarged specimen image, and the growth information can be associated with each other and registered in the growth information database 145. When the first identifier 101 is attached in the vicinity of the microwell 105, the first identifier 101 is further recognized by the processing of the first identifier recognition processing unit 151 described above, and the enlarged specimen image in accordance with the imaging order. It is possible to further determine whether or not For example, in the example of FIG. 8A, when shooting is performed in the wrong shooting order, the well ID recognized by the first identifier 101 is different from the well ID associated with the shooting order. Therefore, the imaging support processing unit 154 determines whether the well ID associated with the imaging order matches the well ID associated with the first identifier 101 recognized by the processing of the first identifier recognition processing unit 151 described above. Further determination may be made. In this way, by combining with the recognition of the first identifier 101, it is possible to determine whether or not the image has been shot without error according to the shooting order.

  In addition, the imaging support processing unit 154 performs a determination process as to whether or not an individual enlarged sample image has been correctly acquired. For example, the imaging support processing unit 154 determines whether or not the enlarged specimen image has been acquired correctly by comparing the number of fertilized eggs recognized from the entire image information with the number of acquired enlarged specimen images. Further, the imaging support processing unit 154 changes the file name according to a file naming rule specified in advance for the image file of the enlarged specimen image. The file naming rule may be set in advance. Further, if the file naming rule is not determined in advance, the shooting support processing unit 154 may display a screen for causing the output unit 133 to input the naming rule and acquire the naming rule from the input information for the screen. . The file naming rule includes at least a character string indicating a cell culture container ID and a character string indicating a well ID. The file naming rule may further include a character string indicating the shooting date and time.

  The feature amount calculation processing unit 155 calculates various feature amounts from the image information of the enlarged specimen image. Below, the feature-value of a human fertilized egg is demonstrated as an example. When cultivating a human fertilized egg, the growth state of the fertilized egg is usually determined in stages after fertilization, thereby determining whether the fertilized egg is of good quality suitable for uterus transplantation.

  First, morphological evaluation performed at the time of the second to third days after the start of culture will be described. There is a method for evaluating the grade of embryos at the time of 3-8 cell stage embryos 2-3 days after fertilization. In this evaluation, the Veek classification is generally used. In the Veek classification, embryos with equally divided cells and less fragmentation (cell debris) are considered better.

The grades of Veek are shown below.
Grade 1: Embryos with uniform cell (blastomere) morphology and no fragmentation.
Grade 2: Embryos with uniform cell (blastomere) morphology but no slight fragmentation.
Grade 3: Embryo with nonuniform cell (blastomere) morphology. Or embryos that accept a small amount of fragmentation.
Grade 4: Embryos with uniform or non-uniform cell (blastomere) morphology and significant fragmentation.
Grade 5: Embryo with few cells (blastomere) and marked fragmentation.

  Next, the feature amount calculation processing by the feature amount calculation processing unit 155 with reference to the Veek classification will be described. FIG. 16 shows an example of calculating the uniformity of cells (blastomere) from the enlarged specimen image. In FIG. 16, the enlarged specimen image is shown by a simple illustration. Note that the subject of the processing described below is a feature amount calculation processing unit 155.

  FIG. 16 (a) shows a portion of a cell cut out from the enlarged specimen image in order to easily explain the present technique. Note that the feature amount calculation processing unit 155 may perform the image processing of the enlarged specimen image as it is, or may perform image processing by cutting out only the cell portion of the enlarged specimen image. As described above, the pixel value of the cell portion in the microwell 105 varies, and an edge can be detected by a known method, so that an image of the cell portion can be cut out. First, an edge corresponding to the outline of the blastomere is detected from the enlarged specimen image. For edge detection, a known method such as a Sobel filter, a Laplacian filter, or a Canny filter can be used. Note that the above-described contour line extraction process when recognizing the first identifier 101 may be used, or a process of estimating and interpolating a portion where the contour line is interrupted may be additionally performed. After detecting the edge, a blast ball detection process is performed. As an example of the blast ball detection process, there is a method of detecting a circle using Hough transform. Alternatively, the blastomere may be detected using a known method such as ellipse fitting.

  FIG. 16B shows a portion detected by the blastomere detection process. For easier understanding, the portion detected as a blastomer is shown as a thick line in a form overlapping the enlarged specimen image. As shown in FIG. 16B, for example, a number or ID is temporarily attached to each detected blast ball in order to distinguish them. Here, blastomeres 1 to 4 are detected. Note that the number of blasts detected here can be registered as information 1107 indicating the second feature amount of the growth information database 145.

Next, a feature amount that is an index of uniformity of the blastomere is calculated. Here, the circularity and the diameter will be described. Here, the “circularity” is a value representing the circularity and can be defined by the following equation.
Circularity = 4πS ÷ L ²
S is the area (number of pixels) and L is the perimeter. This circularity indicates that when the value is 1, it is closest to a circle. In the example of FIG. 16B, the circularity can be calculated by using the number of pixels surrounded by the edge of each blast ball as S and using the peripheral length of the edge of each blast ball as L. “Diameter” is the diameter when the blastomer is approximately a circle, the maximum diameter when it is an ellipse, and the maximum length that passes through the center of gravity in other shapes such as a gourd.

  FIG. 16C shows an example in which the circularity and diameter are calculated for each blastomere. In this way, the feature amount may be calculated for each blast ball, and all these feature amounts may be registered as the growth information. From these plurality of feature amounts, a predetermined score for evaluating the growth stage is obtained. It may be calculated. Here, the uniformity (uniformity) score will be described.

As an example, a score indicating uniformity is defined as follows.
Uniformity = (size uniformity) + (deviation from circularity)
Here, the “size matching degree” is a value indicating the variation in the diameter of the blastomere, and the value becomes closer to 0 as the diameters are aligned, and the value increases as there is a variation. . For example, it can be defined by the following formula.
Size equality = variation coefficient = standard deviation ÷ average value In the example of FIG. 16C, the size uniformity is 0.365.

The “degree of deviation from circularity” is a value that considers deviation from the degree of circularity for all blastomeres. The value is close to 0 if each of the spheres is close to a circle, and the value is such that the deviation from the circle. Will grow. For example, it can be defined by the following formula.
Degree of deviation from circularity = 1-average value of circularity In the example of FIG. 16C, the degree of deviation from circularity is 0.25. Finally, the uniformity is “7.9” from the sum of the degree of size uniformity (0.365) and the degree of deviation from the circularity (0.25). In this way, scores calculated from a plurality of feature amounts may be registered in the growth information database 145. The score calculation method described here is an example, and other calculation methods may be used. For example, an important feature amount may be weighted. Moreover, when calculating a score, you may use the feature-value other than having demonstrated here.

  Next, a process for calculating the fragmentation ratio will be described. FIG. 17 shows an example of calculating the fragmentation ratio from the enlarged specimen image. In FIG. 17, the enlarged specimen image is shown by a simple illustration. First, an embryo area is extracted from the enlarged specimen image. Similar to the processing in FIG. 16, edge detection can be used to extract an embryo region. For edge detection, a known method such as a Sobel filter, a Laplacian filter, or a Canny filter can be used. Note that the above-described contour line extraction process when recognizing the first identifier 101 may be used, or a process of estimating and interpolating a portion where the contour line is interrupted may be additionally performed. After the edge detection, an embryo region detection process is performed. As an example of detection processing of an embryo region, there is a method of detecting a circle using Hough transform. Moreover, you may use well-known methods, such as ellipse fitting. FIG. 17B shows a portion detected by the embryo region detection process. For easier understanding, a portion detected as an embryo region is shown as a dotted line in a form overlapping the enlarged specimen image.

  Next, a fragmentation region is extracted. Fragmentation is cell debris (cell waste smaller in size than the blastomere) that is different from the blastomere in the embryo. As shown in the image of FIG. 17A, since the fragmentation region is a region where cell debris is gathered, the fragmentation region is extracted using a region division method. As a method of area division, known methods such as WaterShed, Split and Merge, Mean Shift, and Grabcut can be used. FIG. 17C shows an example in which a fragmentation region is extracted. Finally, the ratio of the fragmentation area to the portion detected as the embryo area is calculated. This ratio can be calculated from the number of pixels corresponding to each portion. In the example of FIG. 17C, the fragmentation ratio is 42%. The feature amount calculated in this way may be registered in the growth information database 145.

  Next, morphological evaluation performed at the time of the 4th to 5th days after the start of culture will be described. There is a method for evaluating the blastocyst grade 4 to 5 days after fertilization, and the Gardner classification is known. A blastocyst is a structure formed in the early stage of embryogenesis after formation of the cleavage space and before implantation. A blastocyst is composed of an inner cell mass and trophectoderm. The inner cell mass is a cell population formed inside a blastocyst and a cell population that will form a fetus in the future. The trophectoderm is a cell population in which the inner cell mass has different properties and will form part of the placenta in the future.

  According to Gardner's classification, the blastocyst development stage is evaluated in 6 stages, the inner cell mass and the trophectoderm are evaluated in 3 stages, and each combination is evaluated. As an example, FIG. 18 shows a stage of a blastocyst, which briefly shows the inner cell mass and trophectoderm. FIG. 19 is an example of each evaluation of the inner cell mass and the trophectoderm. For example, the size of the inner cell mass and the uniformity of the nutrient ectoderm are used as indicators.

  First, the evaluation of the inner cell mass will be described. The evaluation of the inner cell mass includes the size of the inner cell mass, the uniformity of the cells of the inner cell mass, whether the cells of the inner cell mass are in close contact with each other, or whether the number of cells in the inner cell mass is large. The feature amount calculation processing unit 155 calculates various feature amounts of the inner cell mass from the image information of the enlarged specimen image in the following manner. In FIG. 18, the area of the inner cell mass is clearly shown by the outline, but actually, the inner cell mass appears faintly inside the blastocyst in the enlarged specimen image. Therefore, as an example, the blastocyst is finely divided (in units of pixels or in units of several pixels), and it is determined whether each area is an area of an inner cell mass. The area of the inner cell mass can be defined by pixel value information, granularity (variation of pixel values), spatial frequency, or a combination of these information.

  The size of the inner cell mass is determined by the maximum length in the detected area (the maximum length passing through the center of gravity in the area) and the area of the area. The uniformity of the cells of the inner cell mass is determined by the granularity (pixel value variation) in the detected area. Similarly to the blastomere detection process described above, the number of cells in the inner cell mass may be calculated by detecting an edge in the area of the inner cell mass. Furthermore, it may be determined whether the cells of the inner cell mass are in close contact with each other based on the distance between the cells of the inner cell mass in the area (the distance between the center of gravity of each cell).

  The evaluation of the trophectoderm includes the number of cells in the trophectoderm and the uniformity in the trophectoderm. The feature quantity calculation processing unit 155 calculates various feature quantities of the nutrient ectoderm from the image information of the enlarged specimen image in the following manner. First, a region other than the area of the inner cell mass described above is extracted as a region including the nutrient ectoderm. In the extracted area, edge detection processing and cell detection processing (Hough transform, elliptical fitting) are executed in the same manner as described above, and the number of cells in the nutrient ectoderm is calculated. The uniformity within the nutrient ectoderm can be determined by the variation in granularity of each cell detected by the above-described treatment. As another method, the homogeneity in the trophectoderm may be determined by counting the number of cells having a predetermined diameter or less among the cells detected by the above treatment.

  Although the morphological evaluation performed 2-3 days after fertilization and 4-5 days after fertilization was described above, morphological evaluation may be performed in the early stage of fertilization (after about 1 day). For example, morphological evaluation of pronuclear embryos may be used as an initial evaluation method. The pronuclear stage embryo is an embryo that has passed about ten or more hours after the spermatozoon. When the sperm enters the egg, the pronucleus derived from the ovum and the pronucleus derived from the sperm line up side by side. FIG. 20 shows the morphology of the two pronuclei and the nucleolus within the pronucleus. As shown in FIGS. 20A to 20C, there are differences in the size of the pronucleus, the distance between the two pronuclei, the arrangement of nucleolus in the pronucleus, and the like. In fertility treatment, it is known that there are differences in subsequent embryogenesis in terms of the size and arrangement of the pronucleus, and the size, arrangement, and symmetry of the nucleolus in the pronucleus.

  The feature amount calculation processing unit 155 may calculate various feature amounts of the pronuclear stage embryo from the image information of the enlarged specimen image in the following manner. Similarly to the above, edge detection processing and cell detection processing (Hough transform, elliptical fitting) are executed, and cells within a predetermined diameter range are determined as pronuclei. And for the part determined to be the pronucleus, the size of the pronucleus (diameter and area), the brightness of the pronucleus (brightness), the difference in size between the two pronuclei (diameter and area difference), The distance between the two anterior nuclei (the distance between the centers of gravity) is calculated.

  Further, in the same manner, the nucleolus detection process may be executed for the area determined to be the anterior nucleus. Cell detection processing (Hough transform, ellipse fitting) is executed, and cells in a predetermined diameter range (that is, a diameter smaller than a predetermined threshold) are determined as nucleolus. Then, the size (diameter and area) of the nucleolus, the number of nucleolus, the distance between the nucleolus, and the like are calculated. The feature amount calculated in this way may be registered in the growth information database 145.

  The feature amount calculated by the feature amount calculation processing unit 155 has been described above, but is not limited to the above. In the examples described above, evaluation of Veek, Gardner, and pronuclear embryos has been described, but some of these elements may be implemented, or other classifications similar to these may be used. In addition, cell features include the number of cells, cell shape, cell diameter, maximum length through the center of gravity of the cell area, circularity, cell aspect ratio, cell perimeter, cell area, cell Density (brightness and brightness), cell uniformity (size shift, pixel value variation, etc.), percentage of a given cell to the whole (fragmentation percentage, etc.), cell array (distance between cells and multiple And the combination of these).

  Further, the feature amount calculation processing unit 155 may determine the growth stage and pass / fail of the fertilized egg based on the feature amount described above. The feature amount calculation processing unit 155 may refer to the condition 1203 of the determination profile table 146 to determine whether the above-described feature amount satisfies a predetermined condition, and thereby output a grade 1202 that satisfies the condition. The grade 1202 information output here may be registered in the growth information database 145.

  Furthermore, the feature amount calculation processing unit 155 may switch the profile for determining the growth stage of the fertilized egg based on the elapsed time from the culture start date (reference time). FIG. 21 is a diagram illustrating a profile to be applied according to the elapsed time from the culture start date. As shown in FIG. 21, on the first day, the above-described pronuclear stage evaluation is used, on the second to third days, the Veek evaluation is used, and on the fourth to fifth days, the Gardner evaluation is used. You may do it. Since the determination profile table 146 also stores information on the application time (elapsed time from the reference time) 1201, the feature amount calculation processing unit 155 determines the reference time (for example, treatment date and time) and the acquisition time of the enlarged specimen image. By calculating the elapsed time from the above and referring to the determination profile table 146, a profile corresponding to the elapsed time can be applied. Here, the description has been given of switching the profile with the elapsed time from the reference time, but the present invention is not limited to this method. The feature amount calculation processing unit 155 may switch the profile based on the value of the specific feature amount after calculating the feature amount of the cell in the enlarged specimen image. In this case, an item representing a specific feature amount condition may be added to the determination profile table 146. Further, the feature amount calculation processing unit 155 may switch the profile based on the grade of the growth stage. For example, when a certain level of Veek has been reached at the time of the previous image acquisition, a method of switching to a Gardner evaluation profile may be used this time. This is made possible by holding grade information in the growth information database 145.

  The database creation processing unit 156 associates the well ID associated by the imaging support processing unit 154, the cell culture container ID output from the second identifier recognition processing unit 152, and the image information of the enlarged specimen image with the growth information. Store in database 145. Further, the database creation processing unit 156 associates the information on various feature amounts calculated by the feature amount calculation processing unit 155 and the information on the acquisition time of the enlarged specimen image with the cell culture container ID and the well ID, and the growth information database 145. To store. In addition to these, all image information may be stored in the growth information database 145 in association with the cell culture container ID and the well ID.

  Next, the flow of processing in the growth information management apparatus 130 will be described. FIG. 22 is a flowchart for explaining the flow of processing in the growth information management system 1.

  First, a growth information management program is activated on a predetermined information terminal (2201). Next, the second identifier 102 of the cell culture container 100 is read by the second identifier reader 120 (2202). Next, the second identifier recognition processing unit 152 refers to the second identifier correspondence table 143 using the read information of the second identifier 102. Then, the second identifier recognition processing unit 152 outputs the cell culture container ID corresponding to the second identifier 102, and temporarily records the information of the cell culture container ID in a storage device or the like (2203).

  Next, the operator operates the image acquisition device 110 using the input unit 132, and all the images so that all the microwells 105 and all the arrangement information (character strings around the microwells 105) are included in the frame. Information is acquired (2204). Next, the arrangement information of the microwell 105 is recognized from the entire image information by the well arrangement determination processing unit 153 (2205). Specifically, the microwell 105 is detected from all the image information, and a character string around the microwell 105 is recognized by OCR processing or the like.

  Next, the well arrangement determination processing unit 153 determines whether various information recognized from all the image information is within a normal range with respect to the well design information in the well design information table 144 (2206). The determination may be performed, for example, according to items in the well design information table 144. If it is out of the normal range, the process returns to step 2204.

  Next, the well arrangement determination processing unit 153 determines whether or not there are cells in the microwell 105 from all the image information, and calculates the number of fertilized eggs in the entire image information (2207). Next, the well arrangement determination processing unit 153 displays the calculated number of fertilized eggs on the output unit 133 such as a display, and allows the operator to check whether the number is correct (2208). If the number is correct, the process proceeds to the next step 2209 as it is. If the number of fertilized eggs is different, the operator inputs the correct number of fertilized eggs. Here, the flow of calculating the number of fertilized eggs and photographing only microwells with fertilized eggs is described, but the present invention is not limited to this. For example, steps 2207 and 2208 can be omitted. For example, on the assumption that specimen enlarged images of all microwells 105 are acquired, when the presence / absence of a fertilized egg is registered as information 1105 in the growth information database 145, only the determination of whether or not there are cells in the microwell 105 is performed. The information on the presence or absence of a fertilized egg may be finally registered in the growth information database 145.

  Next, the imaging support processing unit 154 causes the output unit 133 such as a display to display a screen for selecting the imaging order and the naming method for the image file of the enlarged specimen image (2209). The photographing support processing unit 154 temporarily records the photographing order and the naming method input via the input unit 132. As described above, the shooting order, the naming method, and the like may be set in advance. In this case, step 2209 can be omitted.

  Next, when the operator operates the image acquisition device 110 to adjust the focus so that the individual microwells 105 fit in the frame (if the first identifier 101 is included, the first identifier 101 is also included). The photographing support processing unit 154 performs guidance according to the designated photographing order (2210). The operator acquires an enlarged specimen image of the individual microwell 105 according to this guide (2211). At this time, information on the imaging time when the enlarged specimen image is acquired is also recorded (2212). Here, steps 2210 to 2212 are repeated n times (number of fertilized eggs).

  Next, the imaging support processing unit 154 determines whether or not the enlarged specimen image has been correctly acquired by comparing the number of fertilized eggs recognized from the entire image information with the number of acquired enlarged specimen images (2213). ). If the correct number of magnified sample images has not been acquired, the process returns to step 2209 to perform imaging again. If the correct number of enlarged specimen images has been acquired, the process proceeds to the next step.

  Next, the imaging support processing unit 154 changes the file name of the image file of the acquired enlarged specimen image according to the designated naming method (2214). Next, the imaging support processing unit 154, based on the imaging order information (specifically, information such as the well ID correspondence table 142), includes all image information, individual enlarged specimen images, cell culture container IDs, and the like. The well ID is associated with the photographing time (2215). If the first identifier 101 is attached in the vicinity of the microwell 105, the first identifier 101 is recognized by the processing of the first identifier recognition processing unit 151 before the step 2215, and the imaging order is changed. It may be further determined whether or not the enlarged sample image is obtained.

  Next, the feature amount calculation processing unit 155 calculates the feature amount of the cell from the enlarged specimen image (2216). The processing flow here will be described later. Next, the database creation processing unit 156 obtains the well ID, the cell culture container ID, the entire image information, the image information of the enlarged specimen image, the information of various feature amounts, and the information of the photographing time of the enlarged specimen image. The associated information is registered in the growth information database 145 (2217).

  FIG. 23 is a flowchart for explaining a specific processing flow of step 2216 of FIG. The subject of the following processing is the feature amount calculation processing unit 155. First, a profile to be used is selected by referring to the determination profile table 146 (2301). For example, a profile corresponding to the elapsed time can be selected by calculating the elapsed time from the operation date and time and the acquisition time of the enlarged specimen image and referring to the application time 1201 of the determination profile table 146.

  Next, a feature amount corresponding to the selected profile is calculated (2302). For example, in the case of the first record in FIG. 12, the feature amounts of the blast ball and fragmentation are calculated as the feature amounts. Next, a score is calculated based on the calculated feature amount (2303). Finally, the grade is determined by referring to the determination profile table 146 (2304). Here, it is determined whether or not the feature amount calculated in step 2203 satisfies the condition 1203 of the determination profile table 146. If the condition is satisfied, the corresponding grade 1202 is output. The score and grade information may then be displayed on the output unit 133 or registered in the growth information database 145.

  As described above, according to the present embodiment, since the well ID is automatically associated with the enlarged specimen image in accordance with the imaging sequence designated in advance, there is no need to manually associate the enlarged specimen image with the growth information. . In addition, since shooting is guided in accordance with a predetermined shooting sequence, the risk of manual matching errors is reduced. Further, not only the well ID but also the cell culture container ID can be associated with the enlarged specimen image, and all the specimen information can be uniquely identified for the enlarged specimen image. In addition, since information on the image acquisition time is also associated with the enlarged specimen image, it becomes easy to manage information regarding the determination of the growth stage of the fertilized egg. In addition, conventionally, an operator who has viewed an enlarged specimen image visually determines the feature amount of a cell and determines the growth stage of a fertilized egg. The feature amount is calculated, and the growth stage of the fertilized egg can be automatically determined based on the feature amount.

  Next, another embodiment of the present invention will be described. FIG. 24 is a diagram showing another form of the growth information database 145. The same components as those in FIG. 11 are denoted by the same reference numerals and description thereof is omitted. From the image acquisition device 110 such as a microscope, it is also possible to acquire depth information when an image is acquired. Therefore, the depth information 1109 may be managed together with the growth information 1103 by adding an item of depth information (depth from the uppermost end of the fertilized egg) 1109 to the growth information database 145.

  By managing the depth information 1109, not only the two-dimensional image information 1104 but also three-dimensional image information can be managed. For example, the image information 1104 is acquired at a plurality of depths for a certain cell (for example, the first to third records in FIG. 24). The growth information management system 1 may create a three-dimensional polygon graphic by laminating a plurality of images corresponding to the plurality of depths (superimposing them in the depth direction). For modeling from a two-dimensional image to a three-dimensional polygon, a method well known to those skilled in the art may be used. Thereby, the three-dimensional information of a fertilized egg can also be managed. Further, by referring to the three-dimensional polygon, it is possible to determine the development stage of the fertilized egg by a three-dimensional display. In this embodiment, the volume of the cell may be calculated using the information of the three-dimensional polygon, and the volume of the cell may be registered in the growth information database 145 as a feature amount. Alternatively, the shape of the entire fertilized egg may be determined from the information of the three-dimensional polygon, and the shape information may be registered in the growth information database 145 as a feature amount.

  In addition, this invention is not limited to embodiment mentioned above, Other various modifications are included. For example, the above-described embodiment has been described in detail for easy understanding of the present invention, and is not necessarily limited to the one having all the configurations described. In addition, a part of the configuration of an embodiment may be replaced with the configuration of another embodiment, and the configuration of another embodiment may be added to the configuration of an embodiment. In addition, it is possible to add, delete, and replace other configurations for a part of the configuration of each embodiment.

  For example, as described above, the constituent elements of the growth information management device 130 may be distributed to a plurality of information processing devices on the network. For example, the growth information management system may include a terminal (first information processing device) connected to the image acquisition device 110 and a server (second information processing device) including a storage device. Various tables, databases, and the like are stored in the terminal or server according to the corresponding processing. As an example, steps 2201 to 2215 in FIG. 22 may be performed by the terminal, and steps 2216 to 2217 may be performed by the server. In this case, in step 2215, the terminal transmits various associated information to the server. Further, the server may receive various types of information from the terminal and perform feature amount extraction and registration processing in the storage device. In another example, the process of associating the shooting order with the well ID may be performed on the server side. In this case, the terminal may transmit the information on the enlarged specimen image together with the information on the imaging order to the server.

  Although the example of a human fertilized egg was demonstrated as the culture | cultivation object above, it is not limited to this. Examples of cells to be cultured include fertilized eggs, egg cells, ES cells (embryonic stem cells), and iPS cells (artificial pluripotent stem cells). An egg cell refers to an unfertilized egg cell, and includes an immature oocyte and a mature oocyte. After fertilization, the fertilized egg increases in number of cells from the 2 cell stage, the 4 cell stage, and the 8 cell stage by cleavage, and develops into a blastocyst through a morula. Fertilized eggs include early embryos such as 2-cell embryos, 4-cell embryos and 8-cell embryos, morulas, blastocysts (including early blastocysts, expanded blastocysts and escaped blastocysts). A blastocyst means an embryo composed of external cells with the potential to form the placenta and internal cell masses with the potential to form embryos. ES cells refer to undifferentiated pluripotent or totipotent cells obtained from the inner cell mass of a blastocyst. An iPS cell refers to a cell having a pluripotency similar to that of an ES cell by introducing several types of genes (transcription factors) into somatic cells (mainly fibroblasts). That is, the cell includes an aggregate of a plurality of cells such as a fertilized egg and a blastocyst.

  Also, the present invention is preferably suitable for culturing mammalian and avian cells, particularly mammalian cells. Mammals refer to warm-blooded vertebrates, eg, primates such as humans and monkeys, rodents such as mice, rats and rabbits, pets such as dogs and cats, and livestock such as cattle, horses and pigs. Is mentioned. The present invention is particularly suitable for culturing human fertilized eggs.

  Further, as described above, each processing unit of the growth information management device 130 may be realized by a program code of software that realizes the function of the embodiment. In this case, a storage medium in which the program code is recorded is provided to the information processing apparatus, and the information processing apparatus (or CPU) reads the program code stored in the storage medium. In this case, the program code itself read from the storage medium realizes the functions of the above-described embodiment, and the program code itself and the storage medium storing the program code constitute the present invention. As a storage medium for supplying such program code, for example, a flexible disk, CD-ROM, DVD-ROM, hard disk, optical disk, magneto-optical disk, CD-R, magnetic tape, nonvolatile memory card, ROM Etc. are used.

  In addition, the information lines in the drawings indicate what is considered necessary for the description, and do not necessarily indicate all information lines on the product. All the components may be connected to each other.

1: Growth information management system 100: Cell culture vessel 101: 1st identifier 102: 2nd identifier 103: Bottom part 104: Side wall 105: Microwell 106: Cell accommodation part 107: Top and bottom mark 110: Image acquisition device 120: 2nd identifier Reading device 130: Growth information management device 131: Storage device 132: Input unit 133: Output unit 141: Sample information database 142: Well ID correspondence table 143: Second identifier correspondence table 144: Well design information table 145: Growth information database 146 : Determination profile table 151: first identifier recognition processing unit 152: second identifier recognition processing unit 153: well arrangement determination processing unit 154: imaging support processing unit 155: feature amount calculation processing unit 156: database creation processing unit

Claims (11)

A cell culture vessel having a plurality of microwells;
A storage means for storing an enlarged specimen image including one microwell, identification information for identifying the microwell, and growth information of cells in the microwell in association with each other;
A registration unit that registers the enlarged specimen image, the identification information, and the growth information in the storage unit in association with each other based on information in which the imaging order of the plurality of microwells is associated with the identification information;
A growth information management system.   The growth information management system according to claim 1, further comprising imaging support means for performing a guide when imaging the enlarged specimen image according to the imaging order.   The apparatus further comprises well arrangement determining means for recognizing arrangement information relating to the plurality of microwells from image information including all of the plurality of microwells and determining whether the arrangement information matches design information of the microwell. The growth information management system according to 1 or 2.   The growth information management system according to claim 1, further comprising a changing unit that changes a file name of the image file of the enlarged specimen image according to a predetermined naming rule. The cell culture container has a plurality of first identifiers attached in pairs for each of the microwells, and the enlarged specimen image includes the pair of the first identifiers and the microwells,
The determination unit according to claim 1, further comprising: a determination unit that determines whether the identification information associated with the imaging order matches the identification information identifying the microwell associated with the first identifier. The growth information management system described in 1. A program for causing an information processing apparatus including at least a calculation means and a storage means to execute growth information management processing for a cell culture container having a plurality of microwells,
In the calculation means,
Based on information in which the imaging order of the plurality of microwells and identification information for identifying the microwell are associated with each other, the enlarged specimen image including one microwell, the identification information, and the growth of cells in the microwell A program for executing a registration process for registering at least information in association with the storage means. In the calculation means,
The program according to claim 6, further causing an imaging support process for performing a guide when imaging the enlarged specimen image in accordance with the imaging order. In the calculation means,
Recognizing arrangement information regarding the plurality of microwells from image information including all of the plurality of microwells, and further executing a well arrangement determination process for determining whether the arrangement information matches design information of the microwell. Item 8. The program according to item 6 or 7. In the calculation means,
The program according to any one of claims 6 to 8, further causing a change process for changing a file name of the image file of the enlarged specimen image according to a predetermined naming rule. The cell culture container has a plurality of first identifiers attached in pairs for each of the microwells, and the enlarged specimen image includes the pair of the first identifiers and the microwells,
The calculation means further causes a determination process to determine whether the identification information associated with the imaging order matches the identification information identifying the microwell associated with the first identifier. The program according to any one of 6 to 9. A first information processing apparatus comprising image acquisition means for acquiring an enlarged specimen image including one microwell from a cell culture container having a plurality of microwells;
A second information processing apparatus comprising storage means for storing the enlarged specimen image, identification information for identifying the microwell, and growth information of cells in the microwell in association with each other;
With
The first information processing device associates at least the enlarged specimen image with the identification information based on information in which the imaging order of the plurality of microwells and the identification information are associated with each other. Sent to the processing unit,
The second information processing apparatus calculates the growth information from the enlarged specimen image, and registers the enlarged specimen image, the identification information, and the growth information in the storage unit at least in association with each other. .

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