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WO2017110005A1 - Procédé de prélèvement d'objet cible - Google Patents

Procédé de prélèvement d'objet cible Download PDF

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Publication number
WO2017110005A1
WO2017110005A1 PCT/JP2015/086442 JP2015086442W WO2017110005A1 WO 2017110005 A1 WO2017110005 A1 WO 2017110005A1 JP 2015086442 W JP2015086442 W JP 2015086442W WO 2017110005 A1 WO2017110005 A1 WO 2017110005A1
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WO
WIPO (PCT)
Prior art keywords
well
cell aggregate
plate
picking
image
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2015/086442
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English (en)
Japanese (ja)
Inventor
伊藤　三郎
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Yamaha Motor Co Ltd
Original Assignee
Yamaha Motor Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Yamaha Motor Co Ltd filed Critical Yamaha Motor Co Ltd
Priority to PCT/JP2015/086442 priority Critical patent/WO2017110005A1/fr
Publication of WO2017110005A1 publication Critical patent/WO2017110005A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/26—Inoculator or sampler
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting

Definitions

  • the present invention relates to a method for picking up a minute object, such as a cell aggregate, carried on a receiving recess of a plate, using a nozzle.
  • a tip having a tip opening for sucking and discharging the object and capable of holding the object
  • the object is, for example, a cell aggregate.
  • the cell agglomerate may be carried on the well of a well plate having wells (accommodating recesses) arranged in a matrix for processing operations such as observation, selection, inspection or culture. After the processing operation, the cell aggregate is aspirated from the well together with the culture medium by the chip arranged to be movable up and down above the well plate.
  • the tip opening of the tip and the cell aggregate of the suction target are aligned with each other over the well plate, the tip is lowered, and the tip opening approaches the cell aggregate. That is, the chip is approached based on the position where the cell aggregate is carried on the well plate (for example, Patent Document 1).
  • the cell agglomerates are not necessarily carried at the center of the well, and may be carried unevenly near the periphery of the bottom of the well.
  • the outer diameter of the chip needs to be smaller than the inner diameter of the well so as to enter the well, but the difference in diameter between the two is not extremely different.
  • An object of the present invention is to provide a method that allows an object to be satisfactorily picked up when an object carried in an accommodation recess of a plate is sucked by entering a nozzle into the accommodation recess.
  • An object pick-up method is a method for picking up an object carried in the housing recess of a plate having a housing recess on an upper surface, and a nozzle having a tip opening for generating a suction force on the upper surface side of the plate.
  • a step of recognizing the feature a step of causing the calculation means to obtain a center coordinate of the receiving recess in a top view based on the recognition information of the shape feature; and a driving means of the nozzle to move the nozzle to the receiving recess. And lowering the nozzle in a state of being positioned on the axis of the center coordinate in the sky.
  • FIG. 1 is a diagram showing a configuration of a cell transfer device to which the present invention is applied.
  • FIG. 2 is a perspective view of the sorting container.
  • FIG. 3 is a top view of the well plate. 4 is a cross-sectional view taken along line IV-IV in FIG.
  • FIG. 5 is an enlarged photograph of the upper surface of the well plate, showing a state where cell clumps are supported.
  • FIG. 6 is a block diagram showing an electrical configuration of the cell transfer device.
  • FIG. 7 is a top view of a well showing a state in which cell clumps are carried at positions deviated from the center at the bottom of the well.
  • FIG. 8 is a cross-sectional view of a well plate showing a comparative example with respect to the present embodiment and showing a state where a chip is approached to the cell aggregate in the state of FIG.
  • FIG. 9 is a top view of a well showing a state in which the well frame is traced in the pickup method of the present embodiment.
  • FIG. 10 is a cross-sectional view of a well plate showing a state in which a chip approaches a cell aggregate in this embodiment.
  • FIG. 11 is a cross-sectional view of a well plate showing a state of sucking cell aggregates.
  • FIG. 12 is a cross-sectional view of a well plate showing a state of photographing cell aggregates.
  • FIGS. 19A to 19C are diagrams showing modified examples of wells.
  • the object is not particularly limited, but in the present embodiment, a cell derived from a living body, particularly a cell aggregate (spheroid) is exemplified as the object.
  • a cell aggregate derived from a living body is formed by aggregation of several to several hundred thousand cells. Therefore, the size of the cell aggregate is various.
  • Cell aggregates formed by living cells are almost spherical, but if some of the cells that make up the cell aggregates are altered or become dead cells, the shape of the cell aggregate is distorted, or The density may be non-uniform.
  • a cell moving device that moves a cell to a predetermined stage is a preferred application of the present invention.
  • the target objects are small electronic parts and mechanical parts, liquids such as organic or inorganic fragments and particles, pellets, eggs used in in vitro fertilization and IVF (in vitro fertilization), small fish such as zebrafish, Plant seeds may be used.
  • FIG. 1 is a diagram showing a configuration of a cell transfer device S to which the object pickup method according to the present invention is applied.
  • a cell moving device S that moves the cell aggregate C (object) between two containers is illustrated.
  • the cell transfer device S includes a sorting container 1 having a well plate 10 (plate), a microplate 4, a camera unit 5 (imaging means), and a head unit 61 on which a chip 6 (part of a nozzle) is mounted. Yes.
  • the sorting container 1 is a container from which the cell aggregate C is moved, stores the medium L, and holds the cell sorting well plate 10 so as to be immersed in the medium L.
  • the well plate 10 is a plate that carries the cell aggregate C, and has an accommodation recess (well 3) capable of individually accommodating the cell aggregate C on the upper surface.
  • the medium L is not particularly limited as long as it does not deteriorate the properties of the cell aggregate, and can be appropriately selected depending on the type of the cell aggregate.
  • Examples of the medium L include basal medium, synthetic medium, eagle medium, RPMI medium, Fischer medium, ham medium, MCDB medium, and serum medium, as well as glycerol and cell banker (Juji Field Co. )) Cell frozen solution, formalin, reagent for fluorescent staining, antibody, purified water, physiological saline and the like.
  • BxPC-3 human pancreatic adenocarcinoma cells
  • RPMI-1640 medium 10% fetal bovine serum FBS (Fetal Bovine Serum)
  • supplements such as antibiotics and sodium pyruvate may be used as necessary.
  • the sorting container 1 has a cylindrical shape, and has a rectangular upper opening 1H on the upper surface side.
  • the upper opening 1H is an opening for loading the cell aggregate C and picking up the selected cell aggregate C.
  • the well plate 10 is disposed below the upper opening 1H.
  • the sorting container 1 and the well plate 10 are made of a translucent resin material or glass. This is because the cell aggregate C carried on the well plate 10 can be observed by the camera unit 5 disposed below the sorting container 1.
  • FIG. 2 is a perspective view of the sorting container 1
  • FIG. 3 is a top view of the well plate 10
  • FIG. 4 is a cross-sectional view taken along the line IV-IV in FIG.
  • the sorting container 1 includes a bottom plate 11, an outer peripheral wall 12, an inner peripheral wall 13, and a top wall 14.
  • the bottom dish 11 is a cylindrical dish member having an upper surface opening that constitutes the bottom of the sorting container 1.
  • the outer peripheral wall 12, the inner peripheral wall 13, and the top wall 14 constitute a lid member that covers the bottom plate 11.
  • the outer peripheral wall 12 is a portion larger in diameter than the side peripheral wall of the bottom plate 11, and the inner peripheral wall 13 is a rectangular tube-shaped portion disposed inside the outer peripheral wall 12.
  • the top wall 14 is a plate member that covers a region other than the upper opening 1 ⁇ / b> H on the upper surface side of the sorting container 1.
  • the inner peripheral wall 13 is a wall that partitions the upper opening 1H, and is inclined so that the opening area gradually decreases from the upper opening 1H downward.
  • the top wall 14 has a work hole 15 formed of a through hole in the vertical direction. Through this working hole 15, operations such as injection of the medium L into the cavity of the sorting container 1, injection of chemicals, liquid absorption or waste liquid of the medium L, and the like are performed. Further, a pipe connection port 16 for adjusting the atmospheric pressure in the cavity of the sorting container 1 is installed on the top wall 14.
  • the well plate 10 includes a plate body 2 and a plurality of wells 3 (accommodating recesses) formed in the plate body 2.
  • the plate body 2 is made of a flat plate member having a predetermined thickness, and has an upper surface 21 and a lower surface 22.
  • the upper surface 21 is provided with a plurality of wells 3 that carry cell aggregates C.
  • the well plate 10 is held at the lower end portion of the inner peripheral wall 13 with the lower surface 22 spaced from the bottom plate 11 of the sorting container 1.
  • the well plate 10 is immersed in the medium L in the sorting container 1. That is, the culture medium L is injected into the sorting container 1 so that the upper surface 21 of the well plate 10 is positioned below the liquid surface of the culture medium L.
  • Each well 3 includes an opening 31, a bottom 32, a cylindrical wall 33, a hole 34, and a boundary 35.
  • the opening 31 is a square opening provided on the upper surface 21 and has a size that allows the tip opening 6H of the sorting chip 6 to enter.
  • the bottom 32 is located inside the plate body 2 and near the lower surface 22.
  • the bottom 32 is an inclined surface that is gently inclined downward toward the center (the center of the square).
  • the cylindrical wall surface 33 is a wall surface extending vertically downward from the opening 31 toward the bottom 32.
  • the hole 34 is a through hole that vertically penetrates between the center of the bottom 32 and the lower surface 22.
  • the shape of the hole 34 is square when viewed from above, and is concentric with the opening 31.
  • the boundary portion 35 is located on the upper surface 21 and serves as an opening edge of each well 3 and is a ridge line that partitions the wells 3.
  • the well 3 may have a round shape, a triangular shape, a pentagonal shape, a hexagonal shape, or the like, and these may be arranged on the plate body 2 in a honeycomb shape, a linear shape, or at random.
  • the well plate 10 may be provided with only one well 3.
  • each well 3 defines an accommodation space 3H that accommodates the cell aggregate C. It is intended that one cell aggregate C is generally accommodated in the accommodation space 3H. Therefore, the well 3 is set according to the size of the target cell aggregate C. However, in the operation of dispensing a cell culture solution containing a large number of cell aggregates C into the sorting container 1, a plurality of cell aggregates C may enter one well 3.
  • the hole 34 is provided in order to allow small cell aggregates and impurities other than the desired size to escape from the accommodation space 3H. Therefore, the size of the hole 34 is selected so that the cell aggregate C having a desired size cannot pass therethrough and a small cell aggregate or impurities other than the desired size are allowed to pass through. As a result, the cell aggregate C to be sorted is trapped in the well 3, while impurities and the like fall from the hole 34 to the bottom plate 11 of the sorting container 1.
  • the microplate 4 is a container to which the cell aggregate C is moved.
  • the microplate 4 is a plate in which a large number of microwells 41 whose upper surfaces are open are arranged in a matrix. In one microwell 41, a necessary number (usually one) of cell aggregates C are accommodated together with the medium L.
  • the microplate 4 is also made of a translucent resin material or glass. This is because the cell aggregate C carried on the microplate 4 can be observed by the camera unit 5 disposed below the microplate 4.
  • the camera unit 5 includes a camera lens 51 and captures an image of the cell aggregate C held on the well plate 10 in the sorting container 1 or the cell aggregate C held on the microwell 41 in the microplate 4. .
  • the camera unit 5 includes an image sensor such as a CCD image sensor.
  • the camera lens 51 forms an optical image of the cell aggregate C on the light receiving surface of the image sensor.
  • the camera unit 5 is arranged below these so that the camera lens 51 faces the lower surfaces of the sorting container 1 and the microplate 4. That is, the camera unit 5 captures an image of the cell aggregate C carried on the sorting container 1 or the microplate 4 from the lower surface side.
  • the camera unit 5 is movable in the horizontal direction between the lower side of the sorting container 1 and the lower side of the microplate 4 along the guide rail 52 as indicated by an arrow X2 in the drawing.
  • the chip 6 is a tube-shaped member having a tip opening 6H, and sucks and discharges the medium L including the cell aggregate C. Specifically, the chip 6 aspirates the cell aggregate C from the well plate 10 of the sorting container 1, more specifically the cell aggregate C carried in the well 3 of the well plate 10 together with the medium L, Discharge into the microwell 41 of the plate 4. Although not shown, the chip 6 aspirates a reagent solution or the like as needed and discharges it into the microwell 41 carrying the cell aggregate C.
  • the head unit 61 includes a head main body 62 and a head 63 (a part of the nozzle).
  • the head body 62 holds the head 63 so as to be able to advance and retreat in the vertical direction, and is movable in the left-right direction along the guide rail 61R as indicated by an arrow X1 in the drawing.
  • the head main body 62 can also move in a direction (front-rear direction) orthogonal to the paper surface of FIG.
  • the head 63 is composed of a hollow rod.
  • the chip 6 is attached to the lower end of the head 63.
  • a piston mechanism is mounted in the hollow portion of the head 63, and suction force and discharge force are applied to the tip opening 6H of the chip 6 by the operation of the piston mechanism.
  • the connection body of the head 63 and the chip 6 constitutes a nozzle.
  • the head main body 62 incorporates a power unit of the piston mechanism, an elevating mechanism for moving the head 63 in the vertical direction, and a power unit thereof.
  • a plurality of cell aggregates C in a state dispersed in a cell culture solution are injected into the sorting container 1 from a dispensing chip (not shown).
  • the dispensing tip aspirates the cell culture solution together with the cell aggregate C from a container that stores a cell culture solution containing a large amount of the cell aggregate C, and holds it in the chip. Thereafter, the dispensing tip is moved to the upper position of the sorting container 1 and accesses the upper surface 21 of the well plate 10 through the upper opening 1H. And the cell aggregate C hold
  • FIG. 5 is an enlarged photograph of the upper surface 21 of the well plate 10, and the state of the upper surface 21 of the well plate 10 after the cell aggregate C is discharged from the dispensing tip is photographed.
  • one or a plurality of cell aggregates C have entered a part of the well 3 arranged in a matrix.
  • wells 3 in which cell clumps C are not carried and wells 3 in which only relatively small impurities are carried.
  • the cell aggregate C shown here is an aggregate of colon cancer cells, and there are various shapes such as those close to a sphere, those close to an ellipse, and those having a bowl shape.
  • the center of the bottom 32 of the well 3A that is, not directly above the hole 34, lies on the bottom 32 at the side of the hole 34.
  • the cross-sectional shape of the well 3 is symmetrical, and the bottom portion 32 has the lowest portion where the central hole portion 34 is perforated, and the taper that becomes higher toward the cylindrical wall surface 33. It has a shape. Even if the bottom 32 has such a shape, the well 3A and the cell aggregate CA show that there are many cases where the cell aggregate C does not land at the center of the bottom 32.
  • a relatively large cell aggregate CB close to one spherical shape is carried therein.
  • the cell aggregate CB is carried near the center of the bottom 32 of the well 3B. It can be said that the carrying state of the cell aggregate CB is ideal.
  • a relatively small cell aggregate CC close to one sphere is supported.
  • This cell agglomerate CC is also unevenly distributed near the periphery of the bottom 32 instead of the center of the bottom 32, and is supported on the well 3C.
  • the well 3D carries two cells, a cell aggregate CD1 close to an ellipse and a cell aggregate CD2 close to a sphere.
  • the cell aggregate CD2 is supported near the center of the bottom 32 of the well 3D, but the cell aggregate CD1 is supported near the periphery of the bottom 32.
  • the cell aggregate C discharged from the dispensing tip does not fit into every well 3 of the well plate 10 one by one, and even if it enters the well 3, the center position of the bottom 32 is placed. It is not always carried by.
  • the tip opening 6H of the chip 6 approaches the accommodation space 3H of the well 3 and sucks the cell aggregate C together with the medium L.
  • the outer diameter of the tip opening 6H portion of the chip 6 needs to be smaller than the dimension of the horizontal cross section of the well 3 so that it can enter the well 3 from the opening 31. Since the size of one side of the opening 31 of the well 3 is set according to the size of the target cell aggregate C, it is generally as small as several hundred ⁇ m.
  • the size of the tip opening 6H is set in accordance with the size of the cell aggregate C, and cannot be made extremely small due to the difficulty of processing. Further, the tip opening 6 needs to have an inner diameter capable of sucking the cell aggregate C. Therefore, there is no significant difference between the outer diameter of the tip opening 6H and the horizontal cross-sectional dimension of the well 3.
  • a method is shown in which the cell aggregate C carried in the well 3 of the well plate 10 can be well picked up (sucked).
  • the main point is how to align the tip opening 6H of the chip 6 with the cell aggregate C serving as a suction target.
  • an image of the well plate 10 carrying the cell aggregate C is captured by the camera unit 5, and information obtained by performing image processing on the obtained image data is vertically downward toward the well 3.
  • the chip 6 is positioned before being lowered.
  • the configuration related to the positioning of the chip 6 will be described.
  • FIG. 6 is a block diagram showing an electrical configuration of the cell transfer device S.
  • the cell moving device S includes a control unit 7 (calculation means) that controls the movement of the head unit 61, the positioning and raising / lowering of the head 63, the suction and discharge operations of the cell aggregate C by the head 63, and the operation of the camera unit 5.
  • the cell moving device S includes a camera axis driving unit 53 as a mechanism for horizontally moving the camera unit 5, a head unit axis driving unit 64 (a part of nozzle driving means) as a mechanism for horizontally moving the head unit 61, and a head 63.
  • a head drive unit 65 (a part of nozzle drive means) and a display unit 66 are provided as a mechanism for moving up and down and a mechanism for performing suction and discharge operations.
  • the camera shaft drive unit 53 includes a drive motor that moves the camera unit 5 along the guide rail 52.
  • the ball screw is laid along the guide rail 52, the camera unit 5 is attached to a nut member screwed to the ball screw, and the drive motor rotates the ball screw forward or backward. This is a mode in which the camera unit 5 is moved to the target position by rotating.
  • the head unit shaft drive unit 64 includes a drive motor that moves the head unit 61 (head body 62) along the guide rail 61R.
  • a preferred embodiment is a mode in which a ball screw and a nut member are provided as in the camera shaft driving unit 53, and the drive motor rotates the ball screw forward or backward.
  • the head main body 62 is moved in two directions XY, the first ball screw (X direction) along the guide rail 61R and the movement attached to the first nut member screwed to the first ball screw are mounted.
  • a second ball screw (Y direction) mounted on the plate is used. In this case, the head main body 62 is attached to a second nut member screwed into the second ball screw.
  • the head drive unit 65 includes a power unit for an elevating mechanism that moves the head 63 described above in the vertical direction, and a power unit (for example, a motor) for driving a piston mechanism assembled in the hollow part of the head 63 formed of a hollow rod. ).
  • a power unit for example, a motor
  • the lifting mechanism moves the head 63 up and down between the lowered position where the head 63 extends downward from the head body 62 and the raised position where most of the head body 62 is accommodated.
  • the power unit of the piston mechanism raises and lowers the piston member arranged in the head 63, thereby generating a suction force and a discharge force at the tip opening 6 ⁇ / b> H of the chip 6 attached to the head 63.
  • the display unit 66 is composed of a liquid crystal display or the like, and displays an image taken by the camera unit 5, an image subjected to image processing by the control unit 7, and the like.
  • the control unit 7 is composed of a microcomputer or the like and functionally includes an imaging control unit 71, an image memory 72, an image processing unit 73 (image processing unit), a coordinate calculation unit 74 (calculation unit), a suction sequence setting unit 75, an axis.
  • a control unit 76 and a head control unit 77 are provided.
  • the imaging control unit 71 controls the imaging operation of the camera unit 5.
  • the imaging control unit 71 controls the operation of causing the camera unit 5 to capture at least the image of the cell aggregate C carried on the well plate 10 and the image of the upper surface 21 of the well plate 10 for the sorting container 1. To do.
  • the imaging control unit 71 controls an operation for causing the camera unit 5 to capture an image of the microplate 4.
  • the image memory 72 includes a storage area and an external storage provided in the microcomputer, and temporarily stores image data acquired by the camera unit 5.
  • the image processing unit 73 performs image processing on the image data captured by the camera unit 5 and stored in the image memory 72. For example, the image processing unit 73 performs processing for recognizing the presence of the cell aggregate C on the well plate 10 based on the image of the well plate 10 after the cell aggregate C has been dispensed, the cell aggregate C A process for recognizing the distribution of the cell aggregate, a process for recognizing the shape of the recognized cell aggregate C, and a process for determining the quality of the cell aggregate C are performed using an image processing technique. Further, the image processing unit 73 executes processing for recognizing the position of the frame (boundary portion 35) of the well 3 on the upper surface 21 based on the image of the upper surface 21 of the well plate 10.
  • the coordinate calculation unit 74 specifies a cell aggregate C as a suction target to be sucked by the chip 6 among the cell aggregates C existing on the well plate 10 based on the image data processed by the image processing unit 73. I do. Further, the coordinate calculation unit 74 performs a process of calculating the center coordinates of the well 3 in which the cell aggregate C of the suction target is accommodated. As shown in FIG. 3, the well 3 of the present embodiment is square when viewed from above. For this reason, the position of the intersection of the diagonal lines of the square is calculated as the center coordinates.
  • the sorting container 1 holding the well plate 10 is set at a predetermined position on the stage where the cell moving device S is installed. Therefore, the position of the well plate 10 on the stage is generally known. Therefore, it is possible to drive the head unit shaft driving unit 64 based on the position coordinate information of the well plate 10 and move the head unit 61 to the sky of the sorting container 1 (well plate 10).
  • the tip opening 6H of the chip 6 enters the well 3 having a small opening size, and the cell aggregate C is sucked. Therefore, very accurate positioning of the chip 6 is required.
  • the position of the well plate 10 varies depending on the deviation at the time of setting, the assembled state to the sorting container 1 and the like. Therefore, after the sorting container 1 is actually set on the stage, that is, after the installation position of the well plate 10 is determined, it is necessary to acquire an image of the well plate 10 and obtain the center coordinates of the well 3.
  • the aspiration sequence setting unit 75 performs the aspiration operation with the chip 6 when the coordinate calculation unit 74 specifies the well 3 that contains the cell aggregate C as the aspiration target and there are a plurality of wells 3 to be aspirated. Process to set. For example, the suction sequence setting unit 75 sets a path (suction sequence) that sequentially passes through the wells 3 to be suctioned so that the moving distance of the chip 6 on the well plate 10 is the shortest. Further, when the head unit 61 includes a plurality of heads 63 (chips 6), the aspiration sequence setting unit 75 detects the wells 3 that can be aspirated at the same time and incorporates them into the aspiration sequence.
  • a path suction sequence
  • the axis control unit 76 controls the operations of the head unit axis driving unit 64 and the camera axis driving unit 53. That is, the axis control unit 76 controls the axis driving units 64 and 53 to move the head unit 61 and the camera unit 5 to predetermined horizontal target positions. The positioning of the head 63 (chip 6) in the vertical direction of the well 3 to be sucked is realized by the control of the head unit shaft driving unit 64 by the shaft control unit 76.
  • the head control unit 77 controls the head drive unit 65.
  • the head control unit 77 controls the power unit for the lifting mechanism of the head driving unit 65 to raise and lower the head 63 to be controlled toward a predetermined target position. Further, the head control unit 77 controls the power unit of the piston mechanism with respect to the head 63 to be controlled, so that the suction force or the tip opening 6H of the chip 6 attached to the head 63 is applied at a predetermined timing. Generate discharge force.
  • the cell aggregate C carried in the well 3 of the well plate 10 is inserted into the well 3 from the upper surface 21 side of the plate body 2 with the chip 6 having the tip opening 6H.
  • a method is adopted in which suction is performed by causing the tip opening 6H to enter and generating a suction force. In this method, it is important how the chip 6 is aligned with the cell aggregate C to be aspirated.
  • the above-described alignment is performed based on the position coordinates on the well plate 10 of the cell aggregate C specified as the suction target.
  • position (which well 3) of the well plate 10 the cell aggregate C discharged from the dispensing tip onto the well plate 10 is supported. Accordingly, an image of the well plate 10 in which the cell aggregate C is sown is obtained, coordinates where the cell aggregate C exists are obtained, the chip 6 is aligned with the coordinates, the chip 6 is lowered, and the tip opening is obtained. 6H is allowed to approach cell aggregate C.
  • the chip 6 is approached on the basis of the position of the cell aggregate C on the well plate 10 as described above, the following problem occurs.
  • FIG. 7 is a top view of the well 3 showing a state in which the cell aggregate C is supported on the bottom 32 of the well 3 at a position unevenly distributed from the center of the bottom 32.
  • the bottom 32 has a quadrangular pyramid shape, and the cell aggregate C is supported between the cylindrical wall surface 33 and the hole 34 on one surface thereof.
  • FIG. 8 is a cross-sectional view of the well plate 10 showing a comparative example with respect to the present embodiment and showing a state in which the tip opening 6H of the chip 6 is approached to the cell aggregate C in the carrying state of FIG.
  • the coordinates of the carrying position of the cell aggregate C of the suction target are obtained. That is, the x-axis coordinate and the y-axis coordinate indicating the position near the center of the cell aggregate C are obtained based on the photographed image of the well 3 in FIG. A vertical axis passing through the intersection of these x-axis coordinates and y-axis coordinates is determined as z-axis coordinates as shown in FIG.
  • the axis of the tip opening 6H of the chip 6 is aligned with the z-axis coordinate above the well plate 10, and the head 63 descends along the z-axis. Will be.
  • the head 63 descends along the z-axis.
  • the tip opening 6H cannot be lowered to a lowered position suitable for individually sucking the cell aggregate C of the suction target.
  • the position of the tip opening 6H indicated by a dotted line is a desirable lowered position for suction.
  • a wall portion that partitions the adjacent wells 3, that is, a boundary portion 35 between the wells 3, and a cylindrical wall surface 33 that continues below the boundary portion 35 are formed. There is a wall. Therefore, the tip opening 6H cannot sufficiently approach the cell aggregate C and cannot suck the cell aggregate C. Further, when the lower end surface of the chip 6 collides with the boundary portion 35, the chip 6 or the well plate 10 or both may be damaged.
  • the chip 6 is caused to perform the suction operation of the cell aggregate C in the following procedure.
  • Step 1 The well plate 10 is imaged by the camera unit 5, and the obtained image of the well plate 10 is subjected to image processing, and the boundary portion 35 (opening edge) that is an individual frame of the well 3 on the upper surface 21 of the well plate 10. ).
  • Step 2 Based on the recognition information of the boundary portion 35, the center coordinates of the well 3 (bottom portion 32) in the top view are obtained.
  • Step 3 Among the wells 3 arranged in a matrix, the well 3 carrying the cell aggregate C as a suction target is specified.
  • Step 4 With the tip 6 positioned on the vertical axis of the central coordinate above the well 3 that carries the cell aggregate C of the suction target, the tip 6 is lowered and the tip opening 6H is set to the well 3 Enter inside.
  • Step 5 A suction force is generated at the tip opening 6H, and the cell aggregate C in the well 3 is sucked into the chip 6.
  • step 1 the camera unit 5 images the well plate 10 in the same manner as the comparative example described above.
  • the axis control unit 76 controls the camera axis driving unit 53 to move the camera unit 5 directly below the sorting container 1.
  • the imaging control unit 71 controls the camera unit 5 to capture an image of the well plate 10 carrying the cell aggregate C.
  • an image focused exclusively on the cell aggregate C is acquired.
  • the necessary image is an image of the boundary portion 35 of the well plate 10
  • an image focused on the upper surface 21 of the well plate 10 is obtained. get.
  • the obtained image data of the well plate 10 is stored in the image memory 72.
  • the image processing unit 73 performs image processing on the image data to detect the boundary portion 35 of the wells 3 arranged in a matrix.
  • the boundary portion 35 is a ridge line portion formed by tapering the upper end portion of the wall portion forming the cylindrical wall surface 33. If the boundary portion 35 has a flat shape, the cell agglomerate C will land and stay on the flat portion, making it difficult to enter the well 3. However, in the case of the sharp boundary portion 35 that forms a ridgeline, the cell aggregate C that settles toward the boundary portion 35 enters one of the adjacent wells 3 across the boundary portion 35. It becomes easy. On the other hand, such a ridge line clearly appears as an edge on the image. Therefore, the image processing unit 73 can detect the boundary portion 35 by applying a known edge detection image processing technique.
  • FIG. 9 is a top view of the well 3 showing a state in which the cell aggregate C is supported on the bottom 32 of the well 3 at a position unevenly distributed from the center of the bottom 32, as in FIG. 7 described above. .
  • FIG. 9 shows a state in which the ridge line of the boundary portion 35 detected by the image processing is traced as the well frame 35A. Such a trace image is displayed on the display unit 66 as necessary.
  • step 2 the coordinate calculation unit 74 performs processing for obtaining the center coordinates of the well 3 (bottom portion 32) based on the recognition information of the well frame 35A (boundary portion 35) obtained in step 1.
  • the process performed by the coordinate calculation unit 74 is a process of setting the x-axis coordinate and the y-axis coordinate so as to obtain the center of the square. Since the hole 34 is located at the center of the bottom 32, the center coordinate formed by the intersection of the x-axis coordinate and the y-axis coordinate is located in the hole 34.
  • step 3 the image processing unit 73 performs image processing on the image data of the well plate 10, (A) Processing for recognizing the presence of cell aggregate C on the image, (A) processing for recognizing the state of accommodation of the recognized cell aggregate C in the well 3; (C) processing for recognizing the shape of the cell aggregate C; (D) By performing a process for determining whether the cell aggregate C is good or not, the well 3 in which the cell aggregate C as a suction target is carried is specified.
  • the well 3 that does not carry the cell aggregate C is removed from the suction target by the process (a).
  • the process (a) is a process for detecting the well 3 (for example, the well 3D in FIG. 5) in which a plurality of cell aggregates C are carried. This process is, for example, a process of determining whether or not a plurality of cell aggregates C recognized as individuals exist in one well frame 35A.
  • the process (c) is a process of synthesizing the 3D shape of the cell aggregate C from a plurality of images obtained by, for example, changing the focus position for one cell aggregate C in one well 3. .
  • a template set in advance and the cell aggregate C obtained by the shape recognition process are matched, and the quality of the cell aggregate C is determined by whether or not a predetermined criterion is satisfied. This is a process of determining.
  • the image used by the image processing unit 73 in the process of step 3 may be image data of the well plate 10 newly acquired for the step 3, or may be acquired in the previous step 1 and stored in the image memory.
  • 72 may be image data stored in 72.
  • an image of the upper surface 21 of the well plate 10 is acquired (step 1), a plurality of images are captured while shifting the focus position of the camera lens 51 in the z direction in order to detect the in-focus position of the upper surface 21. Is done.
  • the image of the well plate 10 may be displayed on the display unit 66 including the processes (c) and (d) or the process (b), and the user may make a visual determination.
  • step 4 the axis control unit 76 controls the head unit shaft driving unit 64 so that the axis of the tip opening 6 ⁇ / b> H of the chip 6 attached to the head 63 passes through the center axis obtained in step 2 ( The head unit 61 is moved so as to be aligned with the z axis). Thereafter, the head controller 77 controls the head driver 65 to lower the head 63. Thereby, the tip opening 6H of the chip 6 approaches the cell aggregate C accommodated in the well 3.
  • FIG. 10 is a cross-sectional view of the well plate 10 showing a state of execution of the step 4.
  • the chip 6 is lowered along the z-axis, and the tip opening 6H enters the accommodation space 3H of the well 3.
  • the carrying position of the cell aggregate C is eccentric from the center of the bottom 32.
  • the tip 6 is not positioned based on the carrying position of the cell aggregate C, but the tip 6 is positioned based on the center coordinates of the well frame 35A.
  • the chip 6 can enter the well 3 without colliding with its periphery.
  • the chip 6 is lowered to a height that does not crush the cell aggregate C and is suitable for the suction of the cell aggregate C.
  • it is lowered to a height at which a predetermined distance d exists between the apex Ct of the cell aggregate C and the tip opening 6H.
  • a predetermined distance d exists between the apex Ct of the cell aggregate C and the tip opening 6H.
  • the predetermined distance d is about 0 to 150 ⁇ m.
  • step 5 the head control unit 77 controls the head driving unit 65 to generate a suction force at the tip opening 6H of the chip 6 attached to the head 63.
  • FIG. 11 is a cross-sectional view of the well plate 10 showing the state of suction of the cell aggregate C by the chip 6. Since the well plate 10 is immersed in the culture medium L, when a suction force is generated in the tip opening 6H, the liquid flow LF1 of the culture medium L toward the tip opening 6H through the opening 31 and the tip 34 toward the tip opening 6H. A liquid flow LF2 is generated. The cell aggregate C together with the medium L is sucked into the tubular passage 60 of the chip 6 by these liquid flows LF1 and LF2. Thereafter, the head controller 77 raises the head 63. Through the series of operations described above, the cell aggregate C as the suction target is picked up from the well plate 10 and held in the chip 6.
  • the camera unit 5 is caused to image the cell aggregate C carried in the well 3, and the image processing unit 73 determines the height of the top Ct of the cell aggregate C based on the obtained image of the cell aggregate C.
  • tip 6 can be set according to the height position of the top part Ct. Therefore, even when there are various types of cell aggregates C having different shapes, the distance between the top Ct of each cell aggregate C and the tip opening 6H of the tip 6 is made constant, that is, the cell aggregate C is sucked. Suction can be performed at a suitable predetermined distance d.
  • FIG. 12 is a view showing a photographing state of the well plate 10 and the cell aggregate C carried on the well plate 10 by the camera unit 5.
  • the camera unit 5 is disposed below the sorting container 1 and images of the well plate 10 and the cell aggregate C are taken from the lower surface 22 side of the well plate 10. Therefore, although not shown in FIG. 1, in the actual cell moving device S, the illumination unit 54 is disposed above the sorting container 1 as shown in FIG. The illumination unit 54 illuminates the well plate 10 from above. This illumination is used as transmitted illumination of the well plate 10.
  • the method shown in FIG. 13A is used, for example, when the cell aggregate C has poor transparency and the top lens Ct cannot be focused on the top Ct under transmitted illumination. Even in such a case, at least the outer peripheral portion Co of the cell aggregate C can be focused.
  • the focus of the camera lens 51 is moved at a predetermined pitch within a certain range in the z-axis direction in the vicinity where the cell aggregate C exists, and a plurality of images are captured. Then, image processing such as edge detection processing is performed on each image, and these edge detection results are combined to recognize the outer peripheral portion Co. Thereby, the outline of the cell aggregate C can be grasped.
  • the position of the apex Ct can be estimated.
  • the cell aggregate C has a shape belonging to a substantially spherical shape.
  • the distorted cell aggregate C is often excluded from the suction target. Therefore, if the radius of the found contour is obtained, the upward bulge height from the height position of the outer peripheral portion Co can be estimated. That is, if the outer peripheral portion Co is located at the approximate center of the width of the cell aggregate C in the z-axis direction and has a uniform bulge in the upper and lower portions, the z coordinate of the outer peripheral portion Co corresponds to the height of the contour radius. It can be estimated that the z coordinate to which is added is the z coordinate of the top Ct.
  • the method shown in FIG. 13B is used when, for example, the cell aggregate C has relatively good transparency, and the camera lens 51 can be focused on the top Ct. Also in this case, the focus of the camera lens 51 is moved at a predetermined pitch in a certain range in the z-axis direction in the vicinity where the cell aggregate C exists, and a plurality of images are captured. Then, image processing such as edge detection processing is performed on each image. If the top Ct can be focused (the small circle drawn on the cell aggregate C in FIG. 13B indicates the focused area), the area near the top Ct Can be detected. From the information such as the focal length at this time, the z coordinate of the top Ct can be obtained.
  • the degree to which the chip 6 is lowered during the step of allowing the chip 6 to enter the well 3 can be determined by the height position of the top portion Ct. Therefore, by obtaining the center coordinates of the well frame 35A, not only the x and y coordinates at which the tip opening 6H of the chip 6 does not interfere with the well plate 10 can be determined, but also the height most suitable for sucking the cell aggregate C. The tip opening 6H can be lowered to a position.
  • the well 3 to be sucked is set so that the moving distance of the tip 6 on the well plate 10 is minimized or the time required for suction is minimized. It is desirable to set a route (suction sequence) that passes sequentially.
  • the head unit 61 uses a head on which only one head 63 (one chip 6) is mounted or a head on which a plurality of heads 63 (a plurality of chips 6) are mounted. Can do.
  • the suction sequence setting unit 75 determines the suction sequence so that the cell aggregate C determined as the suction target can be efficiently picked up using one or a plurality of chips 6.
  • the order of suction is determined for the plurality of wells 3. Then, according to the aspiration sequence, the chip 6 aspirates the cell aggregate C in the well 3 at the first aspiration point, and moves it to the microplate 4. Subsequently, the cell agglomerate C is sucked back to the well 3 at the second suction point. Hereinafter, this operation is repeated.
  • the well 3 that performs suction in one batch of suction operation is specified.
  • the head unit 61 includes eight heads 63, eight wells 3 to be suctioned are specified by one batch of suction. Further, it is determined by which of the eight heads 63 (chips 6) each cell aggregate C of one batch of eight wells 3 is sucked. When all the wells 3 to be sucked cannot be sucked in one batch, the well 3 to be sucked is specified by the suction operation of the next batch, and which chip 3 is directed to which well 3 is specified.
  • the head unit 61 moves between the well plate 10 and the microplate 4 in batch units.
  • suction is performed on the premise that a suction approach is performed on one well 6 with one chip 6 for each moving unit of the head unit 61.
  • a sequence is set. For example, the first movement coordinate for moving the chip 6 mounted on the first head 63 to the xy coordinate corresponding to the center coordinate of the well 3 to be first suctioned, the chip mounted on the second head 63
  • the movement coordinates of the head unit 61 are determined as a suction sequence, such as the second movement coordinates for moving 6 to the xy coordinates of the well 3 to be sucked second.
  • FIG. 14 is a diagram for explaining an example of simultaneous suction by a plurality of chips 6.
  • the wells 3 of the well plate 10 are arranged at a pitch A in the x direction.
  • the head unit 61 includes three heads 63A, 63B, and 63C, and chips 6A, 6B, and 6C are attached thereto, respectively.
  • cell aggregates C1, C2, and C3 are supported in the well 3 group of the line X1 extending in the x direction.
  • the pitch in the x direction of the well 3 on which the cell clumps C1, C2, and C3 are carried matches the pitch of the chips 6A, 6B, and 6C. Therefore, for example, if the chip 6A is aligned with the center coordinates of the well 3 that contains the cell aggregate C1, the chips 6B and 6C are also aligned with the well 3 that stores the cell aggregate C2 and C3. . That is, the cell aggregates C1, C2, and C3 can be simultaneously sucked with the chips 6A, 6B, and 6C. Similarly, in the group of wells 3 in the line X2, the cell aggregates C4 and C5 can be simultaneously sucked with the chips 6A and 6C.
  • the suction sequence setting unit 75 detects the wells 3 that can be simultaneously sucked by the plurality of chips 6 as illustrated in FIG. If there are wells 3 that can be sucked simultaneously, the suction sequence is set so that these wells 3 are sucked simultaneously.
  • the cell agglomerates C1, C2, and C3 in the line X1 are simultaneously aspirated in the first batch, and the cell agglomerates C4 and C5 in the line X2 are simultaneously aspirated in the second batch.
  • a suction sequence is set such that the remaining chip 6B sucks the cell aggregates in the other wells 3.
  • [Description of operation flow of cell transfer device] 15 and 16 are flowcharts showing an example of the operation of the cell transfer device S.
  • the control part 7 is made to inject
  • the axis control unit 76 controls the camera axis driving unit 53 to move the camera unit 5 to the imaging position below the sorting container 1 (step S2; see FIG. 1).
  • the imaging control unit 71 controls the camera unit 5 to capture an image of the well plate 10 carrying the cell aggregate C. What is acquired here is an image focused on the upper surface 21 of the well plate 10 (step S3). At this time, in order to detect the in-focus position of the upper surface 21, a plurality of images are taken while shifting the focus position of the camera lens 51 in the z direction. These image data are stored in the image memory 72.
  • step S3 The image data obtained in step S3 is read from the image memory 72, and the image processing unit 73 performs image processing such as edge detection. Thereby, the ridgeline of the boundary part 35 of the well 3 is traced as the well frame 35A (step S4).
  • the control unit 7 causes the display unit 66 to display an image of the well plate 10 in which the well frame 35A is traced (step S5).
  • the image processing unit 73 performs image processing on the image of the well plate 10 acquired in step S3.
  • the image processing unit 73 recognizes the cell aggregate C carried on the well plate 10, selects the cell aggregate C as a suction target, and performs a process of specifying the well 3 on which the cell aggregate C is carried. (Step S6).
  • the control unit 7 may accept the designation of the well 3 carrying the cell aggregate C visually selected by the user based on the image of the well plate 10 displayed on the display unit 66. .
  • step S7 the coordinate calculation unit 74 performs numbering on the well 3 specified in step S6 (step S7). This numbering does not indicate the order of suction, but is simply a process of assigning an identification code to the target well 3 that carries the cell aggregate C that is the suction target. Then, the coordinate calculation unit 74 performs processing for obtaining center coordinates (xy coordinates) for the target well 3 based on the information of the well frame 35A obtained in step S4 (step S8). Further, the image processing unit 73 executes processing for recognizing the height position (z coordinate) of the apex Ct of the cell aggregate C accommodated in the target well 3 (step S9). The process of step S9 may be executed between step S5 and step S6.
  • FIG. 17 is a flowchart showing the recognition process of the apex Ct of the cell aggregate C.
  • the imaging control unit 71 controls the camera unit 5 to perform an imaging operation so as to acquire an image focused on the outer peripheral portion Co of the cell aggregate C (step S91).
  • the imaging control unit 71 performs an imaging operation so as to obtain an image focused on the apex Ct of the cell aggregate C (step S92).
  • the image processing unit 73 performs image processing on the image data obtained in step S91 and step S92, and determines whether or not an in-focus image is acquired on the top Ct (step S93). If there is an image focused on the top Ct (YES in step S93), the z coordinate indicating the height position of the top Ct is acquired from information such as the focal length when the image is captured.
  • the height position of the outer peripheral portion Co is determined from information such as the focal length when the image focused on the outer peripheral portion Co is captured.
  • the z coordinate shown is acquired (step S95).
  • the image processing unit 73 estimates the shape of the cell aggregate C of the suction target, and an estimated value of the z coordinate indicating the height position of the apex Ct is obtained from the estimated shape (step) S96).
  • the suction height of the tip opening 6H of the tip 6, that is, the z coordinate for lowering the tip opening 6H is determined (step S97).
  • step S10 it is confirmed whether or not the entire surface of the well plate 10 has been imaged. This is because the entire surface of the well plate 10 may not be captured at one time at the angle of view of the camera lens 51 of the camera unit 5. If the entire surface has not been imaged (NO in step S10), the process returns to step S2 and is repeated. If the entire surface has been imaged (YES in step S10), the process proceeds to step S11.
  • the suction sequence setting unit 75 performs a process of calculating the mutual positional relationship of the target wells 3 numbered in step S7, particularly the pitch between the target wells 3 (step S11).
  • the suction sequence setting unit 75 performs a process of detecting the wells 3 that can be simultaneously sucked by the plurality of chips 6 in one movement unit (step S12).
  • the suction sequence setting unit 75 sets a suction sequence based on the mutual positional relationship of the target wells 3 and information on the wells 3 that can be simultaneously sucked (step S13).
  • the shaft control unit 76 controls the head unit shaft driving unit 64 to move the head unit 61 over the sorting container 1 (step S14).
  • the axis of the tip opening 6H of the tip 6 designated to perform the first suction is positioned on the vertical line of the center coordinates (xy coordinates) of the well 3 designated as the first suction target in the suction sequence. (Step S15).
  • the head control unit 77 controls the head driving unit 65 to lower the head 63 on which the designated chip 6 is mounted (step S16). At this time, the z coordinate determined in step S9 is used. Thereafter, the head controller 77 controls the head driver 65 to generate a suction force at the tip opening 6H of the chip 6 mounted on the lowered head 63. Thereby, the cell aggregate C is sucked into the chip 6 (step S17). This operation is performed simultaneously for the other chips 6 for one batch.
  • the shaft control unit 76 controls the head unit shaft driving unit 64 to move the head unit 61 that has finished the suction operation for one batch to the sky of the microplate 4 to which the cell aggregate C is moved. (Step S18). Thereafter, an operation of discharging the cell aggregate C held on each chip 6 onto the microplate 4 is executed. Then, it is confirmed whether or not the suction sequence is completed (step S19). If the suction sequence has not been completed (NO in step S19), the process returns to step S15, and the suction operation for the next batch is executed. When the suction sequence is completed (YES in step S19), the process is finished.
  • FIG. 18 is a top view showing a well plate 101 according to a modified example.
  • the well plate 101 has a plurality of wells 301 having a hexagonal shape when viewed from above, and the wells 301 are arranged in a honeycomb shape. According to the well plate 101, the area efficiency of the array of the wells 301 can be improved. Further, since the hexagonal well 301 is provided, there is an advantage that the accommodating property of the cylindrical chip 6 is good.
  • FIGS. 19A to 19C are top views of wells according to modifications.
  • the well 302 in FIG. 19A is circular
  • the well 303 in FIG. 19B is a pentagon
  • the well 304 in FIG. 19C is a triangle.
  • the wells of the well plate 10 may be wells 302, 303, and 304 having such shapes.
  • An object pick-up method is a method for picking up an object carried in the housing recess of a plate having a housing recess on an upper surface, and a nozzle having a tip opening for generating a suction force on the upper surface side of the plate.
  • a step of recognizing the feature a step of causing the calculation means to obtain a center coordinate of the receiving recess in a top view based on the recognition information of the shape feature; and a driving means of the nozzle to move the nozzle to the receiving recess. And lowering the nozzle in a state of being positioned on the axis of the center coordinate in the sky.
  • This method includes a step of recognizing the shape feature of the receiving recess and a step of obtaining the center coordinates of the receiving recess. Then, the nozzle is lowered on the axis of the obtained center coordinate. In other words, the nozzle is not lowered with reference to the position where the object is carried, but the nozzle is lowered with reference to the center position in the top view of the housing recess. Therefore, it is possible to reliably lower the nozzle into the housing recess.
  • the step of lowering the nozzle is a step of entering the nozzle into the housing recess, and after the nozzle enters the housing recess, a suction force is generated at the tip opening.
  • the method further includes a step of sucking the object into the nozzle. According to this method, the object can be reliably sucked.
  • the shape feature is a shape of an opening edge of the housing recess. According to this method, since the opening edge easily exhibits a distinct feature in shape, there is an advantage that it is easy to detect the center position of the housing recess in the top view.
  • the image pickup unit picks up the image of the object carried in the housing recess, and the image processing unit has the height of the top of the object based on the obtained image of the object. It is preferable that the method further includes a step of recognizing a position, and in the step of entering, the degree of lowering the nozzle is determined by the height position of the top portion.
  • the lowering degree of the nozzle can be set according to the height position of the top of the object. Therefore, even when there are various objects having different shapes, the distance between the top of each object and the tip opening of the tip is made constant, that is, a predetermined distance suitable for suction of the object is placed, Suction can be performed.
  • the plate is made of a translucent member, and the imaging unit images the plate from the lower surface side of the plate.
  • the nozzle is arranged on the upper surface side of the plate, and the imaging means is arranged on the lower surface side of the plate. For this reason, the nozzle and the imaging means do not interfere with each other, and a series of pickup operations can be executed efficiently.
  • a plurality of the receiving recesses are arranged in a matrix on the upper surface of the plate, and the receiving recesses that carry an object to be a suction target are recognized among the receiving recesses. It is desirable to further include a step of causing
  • the objects are simultaneously picked up by the plurality of nozzles. It is desirable to further include a step of specifying a plurality of accommodating recesses that can be sucked.
  • a combination of a plurality of nozzles capable of simultaneous suction and a plurality of receiving recesses is grasped in advance. Therefore, even when there are many objects to be suction targets, the time required for the suction can be shortened.
  • the object is a cell and the plate is immersed in a medium.
  • the present invention can be applied to medical and biological research applications.
  • the object carried in the accommodation recess of the plate when the object carried in the accommodation recess of the plate is sucked and picked up, the object can be sucked by reliably entering the nozzle into the accommodation recess. Therefore, the pickup failure rate in the object pick-up operation can be significantly reduced.

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Abstract

La présente invention concerne un procédé de prélèvement d'objets cibles, par lequel une extrémité (6) présentant une ouverture d'extrémité (6H) est insérée à l'intérieur d'un puits (3) d'une plaque de puits (10) soutenant un objet cible (C) et y réalise une aspiration, ledit procédé comprenant : l'étape par laquelle la plaque de puits (10) est capturée par un moyen d'imagerie (5) et un cadre de puits (35A) du puits (3) est reconnu par un moyen de traitement d'image (73) sur la base de cette image ; une étape par laquelle les coordonnées centrales dans une vue de surface supérieure du puits (3) sont découvertes par un moyen de calcul (74) sur la base du cadre de puits (35A) ; et une étape par laquelle l'extrémité (6) est abaissée vers le puits (3), dans un état dans lequel l'extrémité (6) est positionnée au-dessus d'une ligne axiale pour les coordonnées centrales dans un espace situé au-dessus du puits (3).
PCT/JP2015/086442 2015-12-25 2015-12-25 Procédé de prélèvement d'objet cible Ceased WO2017110005A1 (fr)

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CN110591915A (zh) * 2019-09-30 2019-12-20 北京工业大学 一种基于微流控的卵细胞培养装置
CN111699243A (zh) * 2018-02-15 2020-09-22 雅马哈发动机株式会社 摄像系统以及生物对象物移动装置
EP3730601A4 (fr) * 2018-01-31 2021-03-10 Yamaha Hatsudoki Kabushiki Kaisha Procédé et dispositif de déplacement de matière biologique
WO2021111285A1 (fr) * 2019-12-02 2021-06-10 Agilent Technologies, Inc. Manipulation de fluide à l'aide d'un marquage de réceptacle pour positionnement d'aiguille
CN113341001A (zh) * 2020-03-02 2021-09-03 安捷伦科技有限公司 在样品容器转移期间利用信息检测进行样品处理
EP3995211A1 (fr) * 2020-11-10 2022-05-11 National Health Research Institutes Procédé de collecte de cellule unique à haut rendement

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