FISH EGG PROCESSING APPARATUS
20200229404 ยท 2020-07-23
Assignee
Inventors
Cpc classification
C12M1/34
CHEMISTRY; METALLURGY
Y02A40/81
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
B65G47/84
PERFORMING OPERATIONS; TRANSPORTING
International classification
Abstract
[Problem] To provide a fish egg processing apparatus capable of performing predetermined processing on fertilized eggs with higher efficiency, and collecting proper fish eggs.
[Solution] When a housing recess that houses fish eggs one by one is conveyed to a predetermined processing position, and a capillary is raised and lowered to inject a gene solution, the housing recess is imaged from a direction that intersects a depth direction and a conveying-in direction, and an injection operation by the capillary is controlled by determining, along a possibility determination flow, whether to inject a predetermined substance into each of the fish eggs before processing, based on a previously determined condition.
Claims
1-6. (canceled)
7. A fish egg processing apparatus, comprising: a processing water tank to which fish eggs are guided; an aligning and conveying unit that conveys, to a predetermined processing position, a housing recess that houses the fish eggs one by one in the processing water tank; a processing unit that raises and lowers an introduction tube to inject a predetermined substance into each of the fish eggs in the housing recess conveyed to the processing position; an image processing unit that images the housing recess from a direction that intersects a depth direction and a conveying-in direction, in association with the processing water tank; and a possibility determination unit that determines whether to inject the predetermined substance into each of the fish eggs before processing, based on an image obtained through the image processing unit and a previously determined condition, wherein an injection operation of the processing unit is controlled based on a determination result of the possibility determination unit.
8. The fish egg processing apparatus according to claim 7, wherein the predetermined condition includes at least one or both of conditions that: the housing recess is within a predetermined range with respect to the processing unit; and one egg membrane and one embryonic membrane are detected within a predetermined range in the housing recess.
9. A fish egg processing apparatus, comprising: a processing water tank to which fish eggs are guided; an aligning and conveying unit that conveys, from a predetermined processing position, a housing recess that houses the fish eggs one by one in the processing water tank; a processing unit that raises and lowers an introduction tube to inject a predetermined substance into each of the fish eggs in the housing recess at the processing position; a sorting unit that determines a conveyance destination according to a quality of each of the fish eggs; an image processing unit that images the housing recess from a direction that intersects a depth direction and a conveying-out direction, in association with the processing water tank; and a pass/fail determination unit that determines whether each of the processed fish eggs is suitable for collection, based on an image obtained through the image processing unit and a previously determined condition, wherein a sorting operation of the sorting unit is controlled based on a determination result of the pass/fail determination unit.
10. The fish egg processing apparatus according to claim 9, wherein the predetermined substance contains pigment at the time of injection in the processing unit, and the predetermined condition includes at least one or two, or all of conditions that: one egg membrane and one embryonic membrane are detected within a predetermined range in the image; the number of pixels of pigment detected in the egg membrane is within a predetermined range; and a ratio of pigment detected in the embryo among the pigment detected in the egg membrane is equal to or higher than a predetermined value.
11. The fish egg processing apparatus according to claim 7, comprising a storage unit that stores the determination result of the determination unit at the processing position for each pitch while the housing recess is pitch-fed, wherein when the sorting unit located a predetermined number of pitches away from the processing position is controlled, a determination result obtained a predetermined number of pitches before is taken out from the storage unit to control the sorting operation of the sorting unit based on the determination result.
12. The fish egg processing apparatus according to claim 8, comprising a storage unit that stores the determination result of the determination unit at the processing position for each pitch while the housing recess is pitch-fed, wherein when the sorting unit located a predetermined number of pitches away from the processing position is controlled, a determination result obtained a predetermined number of pitches before is taken out from the storage unit to control the sorting operation of the sorting unit based on the determination result.
13. The fish egg processing apparatus according to claim 9, comprising a storage unit that stores the determination result of the determination unit at the processing position for each pitch while the housing recess is pitch-fed, wherein when the sorting unit located a predetermined number of pitches away from the processing position is controlled, a determination result obtained a predetermined number of pitches before is taken out from the storage unit to control the sorting operation of the sorting unit based on the determination result.
14. The fish egg processing apparatus according to claim 10, comprising a storage unit that stores the determination result of the determination unit at the processing position for each pitch while the housing recess is pitch-fed, wherein when the sorting unit located a predetermined number of pitches away from the processing position is controlled, a determination result obtained a predetermined number of pitches before is taken out from the storage unit to control the sorting operation of the sorting unit based on the determination result.
15. The fish egg processing apparatus according to claim 7, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
16. The fish egg processing apparatus according to claim 8, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
17. The fish egg processing apparatus according to claim 9, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
18. The fish egg processing apparatus according to claim 10, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
19. The fish egg processing apparatus according to claim 11, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
20. The fish egg processing apparatus according to claim 12, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
21. The fish egg processing apparatus according to claim 13, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
22. The fish egg processing apparatus according to claim 14, wherein the aligning and conveying unit includes an alignment board in a gear shape provided with the housing recess in the whole circumference, and the image processing unit sets, as a reference point, a relative position at which a front end of the introduction tube is positioned with respect to the housing recess, and automatically acquires a coordinate of a standardized reference point, based on reference points in a plurality of housing recesses imaged while the alignment board is rotated.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DESCRIPTION OF THE EMBODIMENT
[0039] An embodiment of the present invention will be described below with reference to the drawings.
[0040] As illustrated in
[0041] As illustrated in
[0042] According to the present embodiment, a zebrafish egg having substantially a spherical shape with a diameter of approximately 1 mm is used as an example of the fish egg e. Since fish is a vertebrate, and a protein in the form that can be used for drug development is easily obtained by gene introduction, and in particular, zebrafish is known as the type with which the fish eggs e, which are fertilized eggs, are efficiently obtained from the breeding water tank B.
[0043] The configuration of each of components of the gene injection apparatus 1 will be described, below.
[0044] As illustrated in
[0045] As specifically illustrated in
[0046] The vibration conveying unit 4 guides the fish eggs e to the processing water tank 8 by applying predetermined vibration to the second net device 32. The fish eggs e to which the vibration is applied by the vibration conveying unit 4 are conveyed and put into the processing water tank 8 quickly and efficiently.
[0047] Here, as illustrated in
[0048] The processing water tank 8 performs processing on the collected fish eggs e, and is provided with the gene injection unit 5 and the aligning and conveying unit 6 which are processing units that inject a predetermined substance into the fish eggs e. More specifically, the processing water tank 8 includes a pre-processing region 81 in which an internal thickness of the processing water tank 8 is sized so that a plurality of fish eggs e cannot be arranged in parallel to house the fish eggs e before processing, and a post-processing region 82 for housing the fish eggs e to which the predetermined processing has been performed. More particularly, the processing water tank 8 has a substantially T-shape in a plan view in which the pre-processing region 81 in which the internal thickness is sized so that a plurality of fish eggs e cannot be arranged in parallel, and the post-processing region 82 are continuous. In the present embodiment, at least a portion at a processing position P is formed of a transparent material to visually recognize at least the processing position P.
[0049] The aligning and conveying unit 6 aligns the fish eggs e introduced into the pre-processing region 81 so that the gene injection unit 5 easily perform processing. The aligning and conveying unit 6 includes an alignment board 61 shaped in a disc body and provided with a plurality of housing recesses 63 formed at a predetermined pitch around the disc at an equal interval, and an aligning pump 62 that ejects the clean water cw to the alignment board 61. The fish eggs e introduced into the pre-processing region 81 are smoothly guided to the alignment board 61 by the clean water cw from the aligning pump 62 and positioned in each of the housing recesses 63. The alignment board 61 corresponds to a placing unit on which the fish eggs e can be placed. In the present embodiment, as illustrated in
[0050] The gene injection unit 5 injects the gene solution G into each of the fish eggs e conveyed by the aligning and conveying unit 6, and includes a capillary 51 having a substantially needle shape that directly injects a gene into each of the fish eggs e, a syringe pump 52 that supplies a predetermined amount of genes to the capillary 51, and a positioner 53 that positions the capillary 51 and the syringe pump 52 in a vertical direction (Z direction). The positioner 53 also positions the capillary 51 and the syringe pump 52 in XY directions. The capillary 51 corresponds to the introduction tube having a tube shape that can introduce a predetermined substance into each of the fish eggs e at a predetermined timing.
[0051] In the present embodiment, there is provided the selection and collection unit 7 that efficiently collects the fish eggs e into which the gene has been injected, from the aligning and conveying unit 6 to the post-processing region 82.
[0052] The selection and collection unit 7 corresponds to a collection device or a collection tank that collects the processed fish eggs e. The selection and collection unit 7 includes a collection container 71 serving as a collection tank accommodated in the post-processing region 82, a collection pump 72 that is provided near the post-processing region 82 in the pre-processing region 81 and ejects the clean water cw to the alignment board 61, a sorting unit 73 that discharges the fish egg e, to which a gene has not been accurately introduced by the gene injection unit 5 included in the processing device, out of the processing water tank 8 through an NG egg exclusion path, and a guide 75 that is provided to cover the lower half of the alignment board 61 in the processing water tank 8 and guides the fish eggs e from the pre-processing region 81 to an OK egg collection path leading to the post-processing region 82. The OK egg collection path is provided along the guide 75. The collection container 71 according to the present embodiment is provided with a slit 74 in a portion facing the post-processing region 82, and thus, can efficiently introduce the fish eggs e moved from the pre-processing region 81 into the collection container 71. Further, in the present embodiment, a part of the guide 75 is formed to enter into the collection container 71, and thus, the fish eggs e are efficiently guided to the collection container 71.
[0053] The sorting unit 73 includes a water flow urging pump for exclusion, an urging water input and output tube serving as the NG egg exclusion path, and a solenoid valve that opens and closes the tube. When the housing recess 63 housing the eggs e determined as NG are sent to an exclusion position, the solenoid valve is opened, and the eggs e are urged by water flow to be removed from the housing recess 63, and then sent out to the urging water input and output tube.
[0054] Here, in the present embodiment, as illustrated in
[0055] The above-described camera C constitutes an optical axis system for imaging fish eggs in a direction that is orthogonal to a conveyance direction of the housing recess 63 and an elevating direction, and the position of the camera C can be adjusted in the XYZ directions through a driving unit Ea of the control device E illustrated in
[0056] The control device E drives the sorting unit 73 and the positioner 53 through the driving unit Ea, and includes an image processor Eb that performs image processing on an image obtained by capturing inside the housing recess 63, a possibility determination unit Ec that determines whether the each of the fish eggs e is suitable for gene injection, and a pass/fail determination unit Ed that determines through the image processing after the injection whether each of the fish eggs e is properly injected.
[0057] In the possibility determination, an image is captured by the camera when the fish eggs have settled after a predetermined time has elapsed since a stroke operation of the capillary 51 serving as the introduction tube. In the pass/fail determination, the gene solution to be injected is colored with a pigment such as phenol red (red). In the pass/fail determination through the image processing, the colored color is detected to determine pass or fail.
[0058] The image processor Eb includes an edge detection filter unit Eb1 that detects contours of a concave portion, an egg membrane, an embryonic membrane, and the like, based on the brightness, or the like of the image captured by the camera C, a template unit Eb2 that stores reference data on the concave portion, the egg membrane, the embryonic membrane, and the like, and a matching unit Eb3 that performs image matching between the data of the template unit Eb2 and the detected edge. The image processor Eb performs image matching according to requests from the possibility determination unit Ec and the pass/fail determination unit Ed, and returns a result. The camera C illustrated in
[0059] The possibility determination unit Ec defines, as conditions for OK to injection the gene solution, a case where the housing recess 63 of the alignment board 61 is within a predetermined range of the image (condition 1), and as illustrated in
[0060] For the condition 2, a case where no fish egg is placed, a case where the fish egg is dead (see
[0061] The pass/fail determination unit Ed defines, as conditions for an injection result being compatible, a case where one egg membrane e1 (outer membrane) and one embryonic membrane e2 (inner membrane) are each detected within the predetermined range of the image (condition 3), a case where the number of pixels of pigment detected in the egg membrane e1 (hatched portions in
[0062] For the condition 3, a case where a fish egg is damaged by gene injection is excluded, and for the condition 4, a case where the gene solution has not been injected into the embryo and a case where the amount of injection is large or small are excluded, and for the condition 5, a case where most of the gene solution has leaked from the embryo after the injection, a case where the gene solution is injected between the egg membrane e1 and the embryo e2, a case where the gene solution is injected in a position deviated in an optical axis direction, and the like are excluded.
[0063] The possibility determination unit Ec and the pass/fail determination unit Ed illustrated in
[0064] Firstly, in the determination flow, it is determined whether the housing recess 63 has been pitch-fed to the processing position (step S1). In the case of YES, it is determined whether to inject a gene to a fish egg e to be injected with the gene, through the possibility determination unit Ec (step S2). If the determination result is OK, the gene solution is injected by raising and lowering the positioner 53 (step S3). Further, whether the injection result is compatible is determined on the fish egg e into which the gene has been injected, through the pass/fail determination unit Ed (step S4). If the determination result is OK, a determination OK flag serving as a determination result flag is turned on (step S5). Conversely, if the determination result is NO in steps S2 and S4, a determination NG flag serving as the determination result flag is turned on (step S6). The determination result is stored in a storage unit Ee (step S7), and the processing ends.
[0065] On the other hand, in the processing flow illustrated in
[0066] Next, it is determined whether the determination is OK or NG (step Sc). In the case of YES, the fish egg e is caused to flow into the OK egg collection path (step Sd), and in the case of NO, the sorting unit 73 is operated to flow the fish egg e into an NG egg collection path (step Se), and the processing ends.
[0067] Thus, within one tact time, the possibility determination and the pass/fail determination on the fish eggs e at the processing position P and the sorting processing of the previously determined fish eggs at the sorting position Q are performed in parallel. The above-described possibility determination and pass/fail determination include a waiting time for waiting for the settlement of the fish eggs e after the pitch-feeding of the alignment board 61 and the operation of the positioner, an image capturing time, an image processing time for determination, and the like.
[0068] It is noted that in the alignment board 61 including the housing recess 63, an error in assembling to a motor is inevitable, and thus, a shaft center of the motor may not coincide with a shaft center of the alignment board 61. Thus, when the alignment board 61 is rotated, the position of the housing recess 63 arranged at the processing position P is not necessarily fixed even if the housing recess 63 can be processed highly accurately to maintain the same shape as that of another housing recess 63. The alignment board 61 pitch-feeds at a predetermined angle from the once determined initial position and does not perform feedback control for positioning the housing recesses 63 at the processing position P every time the alignment board 61 pitch feeds, and thus, even though the capillary 51 is accurately positioned, lowered, and raised, it is inevitable that the housing recesses 63 that have reached the processing position P are slightly displaced from each other.
[0069] Therefore, the image processing unit GP is configured to set, as a reference point X, a relative position at which a front end 51a of the capillary 51 to be positioned with respect to the housing recess 63, and automatically acquire a coordinate of a standardized reference point X, based on reference points X in the plurality of housing recesses 63 imaged while the alignment board 61 is rotated.
[0070] Specifically, in the present embodiment, an edge shape of the housing recess 63 is stored in advance as template data, and a reference point is set to a front end position at which the capillary 51 is to be introduced in the housing recess 63. Next, a process of acquiring the coordinate of the reference point X in the imaged housing recess 63 is performed for all or some of the housing recesses 63 for one round, through an image matching between the housing recess 63 imaged as illustrated in
[0071] As described above, the gene injection apparatus 1 being a fish egg processing apparatus according to the present embodiment includes the processing water tank 8 to which fish eggs are guided, the aligning and conveying unit 6 that conveys, to the predetermined processing position P, the housing recess 63 that houses the fish eggs e one by one in the processing water tank 8, the gene injection unit 5 serving as a processing unit that raises and lowers the capillary 51 serving as an introduction tube to inject the gene solution G as a predetermined substance into the fish eggs e in the housing recess 63 conveyed to the processing position P, the image processing unit GP that images the housing recess 63 from the direction that intersects the depth direction and the conveying-in direction, -in direction in association with the processing water tank 8, and the possibility determination unit Ec that determines whether to inject the predetermined substance into each of the fish eggs before processing, based on an image obtained through the image processing unit GP and a previously determined condition. The injection operation of the gene injection unit 5 is controlled based on the determination result of the possibility determination unit Ec.
[0072] Thus, even if the in-line system is employed, it is possible to prevent the processing from being performed in an improper state not satisfying the conditions or being performed on an improper fish egg.
[0073] Further, the predetermined condition includes conditions that: the housing recess 63 is within a predetermined range with respect to the gene injection unit 5; and one egg membrane and one embryonic membrane are detected within a predetermined range in the housing recess, and thus, it is possible to reliably exclude a case where the housing recess is not correctly positioned or a case where the fish egg is not placed, from fish eggs to be processed.
[0074] Alternatively, the gene injection apparatus 1 being a fish egg processing apparatus according to the present embodiment includes the processing water tank 8 to which fish eggs are guided, the aligning and conveying unit 6 that conveys, from the predetermined processing position P, the housing recess 63 that houses the fish eggs e one by one in the processing water tank 8, the gene injection unit 5 serving as a processing unit that raises and lowers the capillary 51 serving as an introduction tube to inject a predetermined substance into each of the fish eggs e in the housing recess 63 at the processing position P, the sorting unit 73 that determines a conveyance destination according to the quality of the fish eggs e, the image processing unit GP that images the housing recess 63 from the direction that intersects the depth direction and the conveying-out direction, in association with the processing water tank 8, and the pass/fail determination unit Ed that determines whether each of the processed fish eggs is suitable for collection, based on an image obtained through the image processing unit GP and a previously determined condition. The sorting operation of the sorting unit 73 is controlled based on the determination result of the pass/fail determination unit Ed.
[0075] Thus, even if the in-line system is employed, it is possible to remove fish eggs to which the predetermined substance is injected improperly without satisfying the conditions.
[0076] In addition, the gene solution contains pigment at a time of injection in the gene injection unit5, and the predetermined condition include conditions that: one egg membrane e1 and one embryonic membrane e2 are detected within a predetermined range in the image; the number of pixels of pigment detected in the egg membrane e1 is within a predetermined range; and the ratio of pigment detected in the embryo e2 among the pigment detected in the egg membrane e1 is equal to or higher than a predetermined value, and thus, it is possible to reliably exclude a fish egg damaged by the injection, a fish egg of which injection state is improper, or a fish egg from which the injected substance is leaked out.
[0077] The gene injection apparatus 1 includes the storage unit Ee that stores the determination result of the determination units Ec and Ed at the processing position P for each pitch while the housing recess 63 is pitch-fed, and, when the sorting unit 73 located a predetermined number of pitches away from the processing position P is controlled, takes out, from the storage unit Ee, a determination result obtained a predetermined number of pitches before to control the sorting operation of the sorting unit 73 based on the determination result.
[0078] Thus, even if the processing position P is separated from the sorting position Q, the determination process and the sorting process can be performed in parallel.
[0079] Further, the aligning and conveying unit 6 includes the alignment board 61 in a gear shape provided with the housing recess 63 in the whole circumference. The plurality of housing recesses 63 are imaged by the image processing unit GP, and the reference point X is standardized (averaged), in which the reference point X is a relative position of the capillary front end 51a and is set as a preferable position in each of the housing recesses 63.
[0080] Thus, the influence due to assembling errors of the alignment board 61 is minimized, and even if the predetermined substance is injected by raising and lowering the introduction tube while the housing recess is pitch-fed in an initially positioned state, it is possible to effectively avoid a situation where poor injection occurs or a needle breaks as a result of contact with the alignment board.
[0081] Although the embodiment of the present invention has been described above, the present invention is not limited to the configuration of the above-described embodiment. For example, in the above-described embodiment, both the possibility determination and the pass/fail determination are performed, but this does not preclude a simplified configuration to perform only the possibility determination or a simplified configuration to perform only the pass/fail determination.
[0082] Although the mode for introducing a gene into a fish egg is disclosed in the above-described embodiment, it is obviously possible to adopt a mode for injecting, into a fish egg, different substances from genes, for example, cells such as human cancer cells, drugs, drug candidate substances, chemical substances such as toxic substances, or food additives such as seasonings or coloring agents. Further, in the above-described embodiment, a zebrafish egg is applied as a fish egg, but, of course, an egg of another fish may be employed. Further, the specific arrangement of individual constituent components, including a detailed mode such as specific water flow in the processing water tank and other configurations, can be variously modified within the scope of the present invention.
INDUSTRIAL APPLICABILITY
[0083] The present invention can be used as a fish egg processing apparatus for performing predetermined processing mainly on fish eggs.
REFERENCE SIGNS LIST
[0084] 1 . . . Fish egg processing apparatus (gene injection apparatus) [0085] 5 . . . Processing unit (gene injection unit) [0086] 6 . . . Aligning and conveying unit [0087] 8 . . . Processing water tank [0088] 51 . . . Introduction tube (capillary) [0089] 51a . . . Front end [0090] 63 . . . Housing recess [0091] 73 . . . Sorting unit [0092] e . . . Fish egg [0093] e1 . . . Egg membrane [0094] e2 . . . Embryonic membrane [0095] Ec . . . Possibility determination unit [0096] Ed . . . Pass/fail determination unit [0097] Ee . . . Storage unit [0098] GP . . . Image processing unit [0099] X . . . Reference point