阅读说明：本技术 细胞移动时的预处理方法和细胞移动装置 (pretreatment method for cell movement and cell movement device ) 是由 井爪洋平 坂本大 于 2018-03-08 设计创作，主要内容包括：一种细胞移动时的预处理方法,是使用具备细胞的拾取单元的细胞移动装置从包含不同尺寸的多个细胞的集合体拾取所需的细胞并使其移动到指定的移动目的地时的预处理方法,其包括：区分步骤,将所述集合体所含的所述多个细胞中的所述所需的细胞根据尺寸区分为至少两种细胞；以及,供给步骤,将进行了所述区分的细胞以各自被区分的状态供给到能够被所述拾取单元拾取的位置。(A pretreatment method for cell movement, which is a pretreatment method for picking up a desired cell from an aggregate including a plurality of cells of different sizes using a cell moving apparatus provided with a cell pickup unit and moving the cell to a predetermined destination, comprising: a distinguishing step of distinguishing the desired cell among the plurality of cells contained in the aggregate into at least two types of cells according to size; and a supplying step of supplying the cells subjected to the differentiation to a position where the cells can be picked up by the pickup unit in a state where the cells are differentiated from each other.)

1. A pretreatment method for cell transfer, which is a pretreatment method for picking up a desired cell from an aggregate including a plurality of cells of different sizes using a cell transfer device provided with a cell pickup unit and transferring the cell to a predetermined transfer destination, the pretreatment method comprising:

A distinguishing step of distinguishing the desired cell among the plurality of cells contained in the aggregate into at least two types of cells according to size; and

A supplying step of supplying the cells subjected to the differentiation to a position where the cells can be picked up by the picking unit in a state where the cells are differentiated from each other.

2. The method of pretreating while moving cells according to claim 1,

The desired cells are cells having a size exceeding a predetermined minimum size,

The pretreatment method further comprises the following steps:

a first removal step of removing cells below the minimum size from the pool.

3. The method of pretreating while moving a cell according to claim 1 or 2,

The desired cells are cells having a size less than a predetermined maximum size,

The pretreatment method further comprises the following steps:

A second removal step of removing cells having the maximum size or larger from the aggregate.

4. the method for pretreating while moving cells according to any one of claims 1 to 3, wherein:

The aggregate is a cell suspension in which a plurality of cells having different sizes are dispersed,

In the distinguishing step, a filter is used that passes cells of a first size without passing cells of a second size larger than the first size.

5. A method of pretreating a cell during its movement according to claim 4 when dependent on claim 3, characterized in that:

In the second removal step, a filter having a mesh size of 100 μm was used,

In the discriminating step, a filter having a mesh size of 70 μm was used.

6. A method of pretreating a cell during its movement according to claim 4 when dependent on claim 3, characterized in that:

In the first removal step, a filter having a mesh size of 40 μm was used,

In the second removal step, a filter having a mesh size of 100 μm was used,

in the discriminating step, a filter having a mesh size of 70 μm was used.

7. a cell moving apparatus used after applying the pretreatment method for moving a cell according to any one of claims 4 to 6 as dependent on claim 3, the cell moving apparatus comprising:

A picking unit picking up the cells;

A moving unit that moves the pickup unit in a predetermined direction;

A sorting unit that sorts the cells to be moved out of the cells supplied in the supplying step; and

an input unit that receives an input of a mesh size of the filter to be used; wherein the content of the first and second substances,

The sorting means performs the sorting based on a sorting criterion predetermined for cells having a size corresponding to the input mesh size,

the picking unit picks up the cells sorted out by the sorting unit,

The moving unit moves the pickup unit that has picked up the cell to a specified movement destination.

8. The cell movement apparatus according to claim 7, wherein:

The input unit further receives an input of at least one of an upper limit and a lower limit of the size of the cell to be moved,

the sorting unit processes the upper limit or the lower limit of the size as an element of the sorting criterion,

The input unit does not receive an input of at least one of the upper limit and the lower limit of the size when the upper limit or the lower limit of the size contradicts the size of the screen opening.

9. The cell movement apparatus according to claim 7, wherein:

In the supplying step, a plurality of dishes capable of holding cells are prepared, the cells are held by the dishes for each of the cells to be differentiated,

A microtiter plate having a plurality of wells for receiving cells is arranged at the specified destination of movement,

The pick-up unit comprises a tip for performing suction and discharge of the cells,

After the cells are aspirated by the tip and moved to the microtiter plate per each dish, the cells are ejected from the tip to the assigned wells of the plurality of wells.

Technical Field

The present invention relates to a preprocessing method for picking up a desired cell and moving the cell to a predetermined destination by using a cell moving apparatus provided with a cell picking-up means, and a cell moving apparatus to which the method is applied.

Background

For example, in applications of medical treatment and biological research, a single cell, a cell aggregate obtained by three-dimensionally aggregating cells, or a cell mass obtained by culturing a cell fragment in a lump (hereinafter, these will be simply referred to as "cell" in the present specification) may be stored in a well of a microtiter plate (micro plate) having wells (wells) arranged in a matrix, and the processing operations such as observation, confirmation of drug efficacy, examination, and culture may be performed. The cells contained in the wells are sorted on a dish having a holding recess capable of containing the cells.

That is, the disk is interspersed with cell groups dispersed in a cell suspension using a dispensing tip. The dispersed cell population herein includes cells of various sizes and shapes. Cells suitable for the processing operation are sorted out from these cells by taking an image of the dish and performing image processing or the like. Selected cells are picked up from the dish by a tip that enables aspiration and expulsion of the cells, and are moved to the microtiter plate and expelled into the wells. For the cell suspension, it is desirable to perform filtration to remove cells of both excessively large and excessively small sizes that are clearly unsuitable for the treatment operation, before they are spread on the dish. Filtration of cancer cells is mentioned in patent document 1.

Properties of cells such as sensitivity to an object to be examined may vary greatly depending on the size of the cells. Therefore, it is desirable to perform the processing job in units of similar sizes, and thus there are many cases where size sorting is required at the time of the pickup. In this case, if the cell suspension subjected to the filtration only to remove the excessively large and small sizes is spread on the dish, it takes time and labor in the sorting operation. That is, although cells having excessively large and small sizes are removed, the size of the cells contained in the cell suspension varies greatly, and even if size sorting is performed by image processing, for example, it takes time to perform sorting processing.

Disclosure of Invention

The invention aims to provide a pretreatment method for effectively moving required cells by using a cell moving device with a cell picking unit and a cell moving device applying the method.

A pretreatment method for cell movement according to an aspect of the present invention is a pretreatment method for picking up a desired cell from an aggregate including a plurality of cells of different sizes using a cell moving apparatus including a cell pickup unit, and moving the cell to a predetermined destination, the pretreatment method including: a distinguishing step of distinguishing the desired cell among the plurality of cells contained in the aggregate into at least two types of cells according to size; and a supplying step of supplying the cells subjected to the differentiation to a position where the cells can be picked up by the pickup unit in a state where the cells are differentiated from each other.

Another aspect of the present invention relates to a cell moving apparatus comprising: a pick-up unit for the cells; a moving unit that moves the pickup unit in a predetermined direction; a sorting unit that sorts the cells to be moved out of the cells supplied in the supplying step; and an input unit that receives an input of a mesh size of the filter to be used; wherein the sorting unit performs the sorting based on a sorting criterion predetermined for cells having a size corresponding to the input mesh size, the picking unit picks up the cells sorted by the sorting unit, and the moving unit moves the picking unit that has picked up the cells to a predetermined destination.

drawings

FIG. 1 is a schematic view of a cell transfer device according to an embodiment of the present invention.

fig. 2 is a diagram showing a cell dispensing operation performed before a cell moving operation by the cell moving device is performed.

Fig. 3 (a) is a plan view of a dish member provided in a sorting container used in the cell transfer device, and fig. 3 (B) is a cross-sectional view taken along line IIIB-IIIB in fig. 3 (a).

Fig. 4 (a) is a perspective view of a microtiter plate used in the cell transfer device, and fig. 4 (B) is a cross-sectional view of the microtiter plate.

FIG. 5 is a block diagram showing an electrical configuration of the cell transfer apparatus.

fig. 6 (a) is a side view of the cell filter, and fig. 6 (B) is a top plan view of the cell filter.

FIG. 7 is a diagram for explaining a cell filtration operation using the cell filter.

FIG. 8 is a diagram showing the procedure of the pretreatment method during cell movement according to the embodiment of the present invention.

FIG. 9 is a schematic diagram showing a conventional cell picking operation.

FIG. 10 is a schematic diagram showing the cell picking operation according to the present embodiment.

FIG. 11 is a diagram showing a procedure of a pretreatment method in moving a cell according to a modification.

FIG. 12 is a diagram showing a preferred example of the operation of picking up cells from the pretreatment at the time of cell transfer.

FIG. 13 is a diagram showing another preferred example of the operation of picking up cells from the pretreatment during cell transfer.

Detailed Description

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. In the pretreatment method for cell migration and the cell migration apparatus according to the present invention, cells derived from a living body, cell masses, cell aggregates (spheroids), or the like are used as objects to be migrated. For example, a cell aggregate derived from an organism is formed by aggregating several to several hundred thousand cells. Therefore, the cell aggregates vary in size. The cell aggregate formed by the living cells is substantially spherical, but if a part of the cells constituting the cell aggregate is degenerated or becomes dead, the shape of the cell aggregate may be deformed or the density may become uneven. In experiments in the field of bio-correlation technology, medicine, a cell moving device is used which picks up usable cell aggregates from a plurality of cell aggregates in various shapes in a dish carried on a sorting table with a tip and moves them to a microtiter plate. In a microtiter plate, various processes such as observation, confirmation of drug effect, examination, and culture are performed on cell aggregates. In the following description, the cell aggregate is referred to as a cell C in a short term in the meaning of including the cell aggregate.

[ Overall Structure of cell transfer device ]

Fig. 1 and 2 are views schematically showing the overall configuration of the cell transfer device S. Here, a cell transfer apparatus S for transferring cells C between three containers is exemplified. The cell moving device S includes: a translucent base 1 having a horizontal mounting surface (upper surface); a camera unit 5 disposed on the lower side of the base 1; and a head unit 6 disposed above the base 1. The sorting vessel 11 having the dish member 2 is placed at the first placement position P1 of the base 1, and the microtiter plate 4 (fig. 1) is placed at the second placement position P2. Further, the dispensing container 14 (fig. 2) is placed at the third placement position P3 on the base 1. The camera unit 5 and the head unit 6 are movable in the X direction and the Y direction (a direction orthogonal to the paper surface of fig. 1 and 2). The head unit 6 includes: a plurality of heads 61 to which tips 12 (pickup units) for sucking and discharging cells C to move the cells are attached; and a dispensing head 63 to which a dispensing tip 13 for sucking and discharging the cells C for dispensing the cells is attached. The head 61 and the dispensing head 63 are movable in the Z direction.

In general, the cell transfer device S sucks a cell suspension LA containing a large number of cells C from the dispensing container 14, spreads the cell suspension LA on the dish 2 of the sorting container 11 (the above is a dispensing operation), sorts usable cells C from the dish 2, sucks the usable cells C, and discharges the cells C to the microtiter plate 4 (wells 41) (the above is a cell transfer operation). Fig. 1 shows a part of the cell moving action performed in the cell moving device S. Fig. 2 is a diagram showing a dispensing operation performed before the cell moving operation of fig. 1 is performed. In this dispensing operation, the pretreatment method for cell movement according to the present invention is applied. Note that, in fig. 1, the dispensing tip 13 is not shown for simplicity of illustration. Next, each part of the cell transfer apparatus S will be described.

The base 1 is a rectangular flat plate having a predetermined rigidity and partially or entirely formed of a translucent material. The base 1 is preferably a glass plate. By forming the base 1 from a translucent material such as a glass plate, the sorting container 11 (dish 2) and the microtiter plate 4 disposed on the upper surface of the base 1 can be imaged via the base 1 by the camera unit 5 disposed below the base 1.

The dispensing container 14 is a container having an open top surface and storing a cell suspension LA containing a large number of cells C. The cell suspension LA contains cells C to be sorted in the sorting container 11 and impurities inevitably mixed therein.

The sorting vessel 11 is a vessel which becomes a moving starting point of the cells C, stores the medium L, and holds the dish member 2 for cell sorting in a state of being immersed in the medium L. The dish 2 is a plate for holding the cells C, and has a plurality of holding recesses 3 on the upper surface thereof, which can individually accommodate and hold the cells C. The medium L is not particularly limited as long as the properties of the cells C are not deteriorated, and can be appropriately selected according to the type of the cells C.

The sorting container 11 has a rectangular upper opening 11H on the upper surface side thereof. The upper opening 11H is an opening for inputting the cells C and picking up the sorted cells C. The dish 2 is disposed below the upper opening 11H. The sorting container 11 and the dish member 2 are made of a translucent resin material or glass. This is to enable the cells C carried on the dish member 2 to be observed by the camera unit 5 disposed below the sorting container 11.

A plurality of cells C dispersed in the cell suspension LA are injected from the dispensing tip 13 into the sorting container 11. The dispensing tip 13 sucks and holds the cell suspension LA from the dispensing container 14 storing the cell suspension LA containing a large amount of cells C at the third placement position P3 in the dispensing tip 13. Thereafter, the dispensing tip 13 moves to the position above the sorting container 11 (first placement position P1), and reaches the upper surface of the disk 2 through the upper opening 11H. Then, in a state where the distal end opening t of the dispensing tip 13 is immersed in the culture medium L in the sorting container 11, the cells C dispersed in the cell suspension LA held in the dispensing tip 13 are discharged onto the dish 2. Here, the cell suspension LA is subjected to filtration (pretreatment) for removing the cells C having an excessively large size and an excessively small size, which are obviously unsuitable for use in an inspection or the like, before being spread on the dish member 2. This point will be described in detail later.

The detailed construction of the dish 2 is explained. Fig. 3 (a) is a plan view of the dish 2, and fig. 3 (B) is a cross-sectional view taken along line IIIB-IIIB of fig. 3 (a). The dish 2 includes a dish body 20 and a plurality of retaining recesses 3 formed in the dish body 20. The dish main body 20 is formed of a flat plate-like member having a prescribed thickness, and has an upper surface 21 and a lower surface 22. The holding recess 3 has a receiving opening (opening 31) for the cell C on the upper surface 21 side. The dish 2 is immersed in the medium L in the sorting vessel 11. Specifically, the upper surface 21 of the dish body 20 is immersed in the culture medium L in the sorting container 11, and the lower surface 22 is held in the sorting container 11 with a space therebetween with respect to the bottom plate of the sorting container 11 (see fig. 1).

Each holding recess 3 includes an opening 31, a bottom 32, a cylindrical wall surface 33, a hole 34, and a boundary 35. In the present embodiment, an example is shown in which the holding recesses 3 having a square shape in a plan view are arranged in a matrix. The opening 31 is a square opening provided in the upper surface 21, and has a size allowing the entry of the tip opening t of the selected nib 12. The base 32 is located on the interior of the dish body 20 adjacent the lower surface 22. The bottom 32 is an inclined surface gently inclined downward toward the center (the center of the square). The cylindrical wall surface 33 extends vertically downward from the opening 31 toward the bottom 32. The hole portion 34 is a through hole vertically penetrating between the center of the bottom portion 32 and the lower surface 22. The boundary portion 35 is located on the upper surface 21, and is a portion that becomes an opening edge of each holding recess 3, and is a ridge line that divides the holding recesses 3 from each other.

The bottom 32 and the cylindrical wall 33 of each holding recess 3 define a housing space 3H in which the cells C are housed. The housing space 3H is generally designed to house one cell C. The holes 34 are provided to release small cells and foreign substances having a size other than a desired size from the housing space 3H. Therefore, the size of the pores 34 is selected to be a size that does not allow the passage of cells C of a desired size but allows the passage of small cells or inclusions other than the desired size. Thus, the cells C to be sorted are captured in the holding pocket 3, while the foreign substances and the like fall from the holes 34 to the bottom plate of the sorting container 11.

Returning to fig. 1, the microtiter plate 4 is a container to which the cells C are transferred, and has a plurality of wells 41 through which the cells C are discharged. The well 41 is a bottomed well that opens on the upper surface of the microtiter plate 4. A desired number (usually 1) of cells C are accommodated in one well 41 together with the medium L. The microtiter plate 4 also uses a member made of a light-transmitting resin material or glass. This is to enable observation of the cells C carried in the wells 41 by the camera unit 5 disposed below the microtiter plate 4.

Fig. 4 (a) is a perspective view showing an example of the microtiter plate 4. The microtiter plate 4 includes a plate body 40 and a plurality of wells 41 arranged in a matrix in the plate body 40. When discharging the cells C, the tip opening t of the tip 12 enters the hole 41, and thus each hole 41 has an opening diameter that allows the tip 12 to enter with a margin.

there are baseline dimensions for commercially available microtiter plates. The reference microtiter plate was provided with the specified longitudinal x transverse dimensions (longitudinal 85.48mm x transverse 127.76mm) with the specified number of wells. The number of wells is generally 24X 16 (384 wells), and the wells are arranged in a matrix at a predetermined pitch.

fig. 4 (B) is a sectional view of the 384-well microtiter plate 4. As shown, 24 wells 41 are arranged at a uniform well pitch (16 in the short side direction) in the long side direction of the microtiter plate 4.

The camera unit 5 captures images of the cells C held in the sorting container 11 or the microtiter plate 4 from the lower surface side thereof, and includes a lens portion 51 and a camera body 52. The lens unit 51 is an objective lens used in an optical microscope, and includes a lens group for forming an optical image of a predetermined magnification and a lens barrel for accommodating the lens group. The camera body 52 includes an image pickup device such as a CCD image sensor. The lens unit 51 forms an image of light of an imaging object on a light receiving surface of the imaging element.

The head unit 6 is provided for moving the cell suspension LA from the dispensing container 14 to the sorting container 11 and for moving the cells C from the disk 2 to the microtiter plate, and includes a plurality of heads 61, a single dispensing head 63, and a head body 62 to which these are assembled. A tip 12 (pick-up unit) for sucking and discharging the cells C is attached to the tip of each head 61. The dispensing tip 13 described above is attached to the tip of the dispensing head 63. The head main body 62 holds the head 61 and the dispensing head 63 so as to be movable up and down in the + Z and-Z directions. As described above, the head unit 6 is movable in the XY direction, and therefore functions as a moving unit that moves the tip 12 and the dispensing tip 13 in a predetermined direction.

The head 61 and the dispensing head 63 are formed of hollow rods to which a negative pressure generating mechanism is attached. For example, a piston mechanism is mounted in the hollow portions of the head 61 and the dispensing head 63, and a suction force and a discharge force are applied to the distal end opening t of the tip 12 or the distal end opening t of the dispensing tip 13 by the operation of the piston mechanism. The head main body 62 incorporates a power unit of the piston mechanism, and an elevating mechanism and a power unit thereof (a head driving unit 65 described below) for moving the head 61 and the dispensing head 63 in the vertical direction.

[ Electrical Structure of cell transfer device ]

FIG. 5 is a block diagram showing the electrical configuration of the cell transfer device S. The cell transfer device S includes a control unit 7 that controls the movement of the head unit 6, the lifting and lowering of the head 61 and the dispensing head 63, the suction and discharge operations of the cells C, the movement and imaging operations of the camera unit 5, and the like. The cell transfer device S includes a camera shaft driving unit 53 as a mechanism for horizontally moving the camera unit 5, a head unit shaft driving unit 64 (moving means) as a mechanism for horizontally moving the head unit 6, a head driving unit 65 as a mechanism for raising and lowering the head 61 and the dispensing head 63, and a mechanism for performing suction and discharge operations, and further includes a display unit 54 and an input unit 55.

The camera shaft driving section 53 includes a driving motor for horizontally moving the camera unit 5 along the guide rail 5G. The preferred mode is as follows: a ball screw is laid along the guide rail 5G, the camera unit 5 is mounted on a nut member screwed with the ball screw, and the driving motor rotates the ball screw in the forward direction or the reverse direction, thereby moving the camera unit 5 to a target position.

The head unit shaft driving section 64 includes a driving motor that moves the head unit 6 (head main body 62) along the guide rail 6G. The preferred mode is as follows: the drive motor rotates the ball screw in a forward direction or a reverse direction. When the head main body 62 is moved in both XY directions, a first ball screw (X direction) along the guide rail 6G and a second ball screw (Y direction) mounted on a moving plate attached to a first nut member screwed to the first ball screw are used. In this case, the head main body 62 is attached to a second nut member (the same applies to the camera shaft driving section 53) screwed to the second ball screw.

The head driving unit 65 is a power unit for the elevating mechanism and a power unit (for example, a motor) for driving the piston mechanism, and is incorporated in the head main body 62. The lifting mechanism moves the head 61 or the dispensing head 63 up and down between a lowered position at which the head 61 or the dispensing head 63 is projected downward from the head body 62 and a raised position at which most of them is accommodated in the head body 62. The power unit of the piston mechanism generates a suction force and a discharge force at the tip opening t of the tip 12 or the dispensing tip 13 by moving up and down a piston member disposed in the head 61 or the dispensing head 63.

The display unit 54 is configured by a liquid crystal display or the like, and displays an image captured by the camera unit 5, an image subjected to image processing or the like by the control unit 7, and the like.

The input unit 55 is constituted by a keyboard, a touch panel, a communication unit for performing data communication with another communication device, and the like, and receives input of operation information and various data from a user. In the present embodiment, the input unit 55 also functions as an input unit that receives an input of the mesh size of the filter used in the sorting step of sorting the desired cells C according to size when the dispensing operation is performed.

The control unit 7 is configured by a microcomputer or the like, and functions to include a shaft control unit 71 (a part of the moving means), a head control unit 72 (a part of the pickup means), an imaging control unit 73, an image memory 74, an image processing unit 75 (a part of the sorting means), a sorting unit 76 (a part of the sorting means), and a storage unit 77 by executing a predetermined program.

The shaft control section 71 controls the operation of the head unit shaft driving section 64. That is, the axis control unit 71 controls the head unit axis driving unit 64 to move the head unit 6 to a predetermined target position in the horizontal direction. The movement of the dispensing head 63 (dispensing tip 13) between the dispensing container 14 and the sorting container 11, the movement of the head 61 (tip 12) between the sorting container 11 and the microtiter plate 4, the positioning in the vertical air with respect to the holding recess 3 of the disk 2, the positioning in the vertical air with respect to the well 41 of the microtiter plate 4 to be discharged, and the like are realized by the control of the head unit shaft drive unit 64 by the shaft control unit 71.

The head control section 72 controls the head driving section 65. The head control unit 72 controls the power unit for the elevating mechanism of the head driving unit 65 to elevate the head 61 or the dispensing head 63 to be controlled toward a predetermined target position. The head control unit 72 controls the power unit of the piston mechanism for the head 61 or the dispensing head 63 to be controlled, thereby generating a suction force or a discharge force at the tip opening t of the tip 12 or the dispensing tip 13 at a predetermined timing.

The imaging controller 73 controls the camera shaft driver 53 to control the movement of moving the camera unit 5 along the guide rail 5G. The imaging control unit 73 controls the imaging operation of the camera unit 5 with respect to the dish 2 or the microtiter plate 4.

The image memory 74 is configured by a storage area, an external memory, and the like provided in the microcomputer, and temporarily stores image data acquired by the camera unit 5.

the image processing section 75 performs image processing on the image data captured by the camera unit 5 and stored in the image memory 74. The image processing unit 75 performs, on the basis of the image of the dish 2 to which the cells C have been dispensed, a process of recognizing the presence of the cells C on the dish 2, a process of recognizing the distribution of the cells C, a process of recognizing the size, shape, color tone, and the like of the recognized cells C on the image, and the like, by using an image processing technique.

the sorting unit 76 sorts the cells C to be transferred, that is, the cells C transferred from the dish member 2 to the microtiter plate 4, based on a predetermined sorting criterion. The object to be sorted is the cell C existing on the dish 2 whose size, shape, color tone, and the like are determined by the image processing unit 75. It is desirable that the size of the mesh of the cell filter 8 to be described later received by the input unit 55 be referred to when the above sorting is performed.

The storage unit 77 stores various setting values and data in the cell transfer device S. In addition, the storage unit 77 stores data on sorting criteria for each size of the cells C. The cell C may have different criteria for quality determination depending on the size thereof, and it is desirable to have a sorting criterion for each size, which is obtained from the result of machine learning, for example. When performing the above sorting, the sorting unit 76 reads the sorting criterion stored in the storage unit 77 and performs a predetermined determination process. The mesh size can be used as a trigger for this readout.

[ operation of cell transfer device ]

Next, a cell transfer method using the cell transfer device S according to the present embodiment will be described with reference to fig. 1 and 2. First, a process of preparing a required device is performed. The sorting container 11, the microtiter plate 4, and the dispensing container 14 are placed at predetermined first to third placement positions P1 to P3 within the movable range of the head unit 6, respectively. The dispensing container 14 is filled with a cell suspension LA subjected to pretreatment (described in detail later) for filtering cells C of a desired size from a cell suspension including cells cultured in a culture container or the like.

First, a dispensing operation of the cells C is performed. The shaft control unit 71 controls the head unit shaft driving unit 64 so that the head unit 6 having the dispensing tip 13 attached to the dispensing head 63 moves above the dispensing container 14, as shown in fig. 2. Next, the head controller 72 controls the head driver 65 to lower the dispensing head 63, and immerse the distal end opening t of the dispensing tip 13 in the cell suspension LA in the dispensing container 14. In this state, the head driving unit 65 generates a suction force in the dispensing head 63, and the cell suspension LA is sucked into the dispensing tip 60.

After that, the dispensing head 63 is raised, and the head unit 6 is moved to the upper position of the sorting container 11. The dispensing head 63 is lowered again, and the tip opening t of the dispensing tip 13 reaches the upper surface 21 of the dish 2 through the upper opening 11H of the sorting container 11. Then, the cell suspension LA held in the dispensing tip 13 is discharged in a state where the tip opening t is immersed in the culture medium L in the sorting container 11. That is, the cells C are scattered on the dish 2.

Subsequently, the sorting operation of the cells C is performed. The imaging controller 73 controls the camera shaft driver 53 to move the camera unit 5 along the guide rail 5G to a position below the sorting container 11. Then, the imaging control unit 73 controls the camera unit 5 to image the cells C (fig. 1) carried in the dish 2. The acquired image data is temporarily stored in the image memory 74, and the image processing unit 75 performs predetermined image processing. Then, the sorting unit 76 determines to sort out the cells C to be moved (cells C of good quality). The sorted cells C are handled as pickup objects of the tip 12, and the coordinate positions thereof are obtained.

Then, a cell movement action is performed. The shaft control section 71 controls the head unit shaft driving section 64 to move the head unit 6 having the tip 12 attached to the head 61 to the upper space of the sorting container 11. Then, the head control section 72 controls the head driving section 65 so that the head 61 is lowered, and the distal end opening t of the pointed end 12 reaches the upper surface of the dish 2 through the upper opening 1H. At this time, XYZ coordinate information indicating the position of the cell C to be moved is supplied to the axis controller 71 and the head controller 72, so that the tip 12 reaches the holding concave portion 3 on which the cell C is carried.

Then, the head driving section 65 generates a suction force in the head 61. Thereby, the target cells C are sucked from the dish 2 (holding recess 3) into the tip 12 together with the medium L (picking up of the cells C). After that, the head 61 is raised, and the head unit 6 is moved to the upper position of the microtiter plate 4 (the pickup unit is moved to a specified movement destination).

When the head unit 6 reaches the upper level of the microtiter plate 4, the head 61 is lowered again until the tip opening t of the tip 12 enters the well 41 of the microtiter plate 4. Then, the head driving part 65 generates a discharging force at the head 61, so that the cells C in the tip 12 are discharged into the holes 41. The discharge state of these cells C is confirmed by imaging the microtiter plate 4 with the camera unit 5.

[ method of pretreatment during cell migration ]

Next, a pretreatment method performed when the desired cells C are picked up from the dish member 2 and transferred to the microtiter plate 4 using the cell transfer device S having the tip 12 as described above will be described. The pretreatment here is as follows: a cell suspension LA containing only cells C having a desired size is filtered from a cell suspension LA that is an aggregate containing a plurality of cells C and inclusions having different sizes, and is supplied to the dispensing container 14.

< cell Filter >

For example, the cell filter 8 shown in fig. 6 is used for the filtration. Fig. 6 (a) is a side view of the cell filter 8, and fig. 6 (B) is a top plan view of the cell filter 8. The cell filter 8 includes a cylindrical frame 81 and a filter membrane 82 attached to the frame 81.

The frame 81 includes a plurality of side ribs 811 as vertical ribs arranged in the circumferential direction, and an annular base rib 812 connecting lower ends of the side ribs 811. A flange portion 83 is provided at the upper end of the side rib 811. An opening 813 is present between the side ribs 811. An opening is also present inside the base rib 812. The filter membrane 82 is a membrane having a mesh which is a fine opening, and is attached to the frame 81 so as to cover the opening 813. Regarding the mesh size of the filter membrane 82, it is selected according to the size of the cells C to be filtered. The operator manipulates the cell filter 8 by using the grip portion 84 provided to protrude from the flange portion 83.

FIG. 7 is a diagram for explaining the operation of filtering the cells C by using the cell filter 8. For filtration, a tube 85 comprising a cylindrical container with an open upper end and a pot 86 for storing the cell suspension LA taken out from the cell culture vessel are prepared. The cell filter 8 is fitted into the tube 85 so that the flange portion 83 is supported by the upper end of the tube 85. Then, the cell suspension LA is poured from the pot 86 through the upper surface opening of the cell filter 8.

Then, the cell suspension LA containing the cells C having a size that can pass through the mesh of the filter membrane 82 is stored in the tube 85. When the cell filter 8 is used for removing unnecessary cells C, the cell suspension LA stored in the tube 85 is directly supplied to the dispensing container 14. On the other hand, if the cell filter 8 is used for the purpose of trapping the desired cells C, the cells C that cannot pass through the filter membrane 82 and are trapped in the cell filter 8 are taken out, dispersed in the cell culture solution, and then supplied to the dispensing container 14.

< specific examples of pretreatment >

FIG. 8 is a diagram showing the procedure of the pretreatment method during cell movement according to the embodiment of the present invention. The pretreatment method includes a first removal step, a second removal step, a separation step, and a supply step. In fig. 8, the state of the cell suspension LA in each step is shown by the open boxes in the stages a1 to a 4. The longitudinal length of each cell represents the variation in the size (diameter) of the cells C contained in the cell suspension LA. Fig. 8 shows that, for example, the cell suspension LA at stage a1 immediately after the culture contains a large number of cells C having a size in the range of more than 0 μm and 250 μm or less.

The cell suspension LA in stage a1 is a cell suspension (corresponding to the "aggregate" of the present invention) in which cells C having a wide variation range of sizes are dispersed and which further contains inclusions and the like. In order to extract cells having a desired size (desired cells) from the cell suspension LA at stage a1, a first removal step and a second removal step are performed. The desired cells are cells C having a size to be examined in the microtiter plate 4, and are cells C having a size exceeding a predetermined minimum size (herein, 40 μm) and smaller than a predetermined maximum size (herein, 100 μm).

In the first removal step, filtration is performed to remove the cells and foreign substances having the minimum size (40 μm) or less from the cell suspension LA in the stage a 1. Therefore, in the first removal step, the cell filter 8 having a mesh opening size of 40 μm is used. Specifically, the cell filter 8 having a mesh size of 40 μm is inserted into the tube 85, and the cell suspension LA of stage a1 is stored in the pot 86.

Subsequently, the cell suspension LA was injected from the pot 86 into the tube 85 equipped with the cell filter 8(40 μm). Then, the cells trapped in the cell filter 8 are removed and dispersed in a cell culture medium or the like to obtain a cell suspension LA in stage a 2. The cell suspension LA stored in the tube 85 is discarded after being filtered by the cell filter 8.

In the second removal step, filtration is performed to remove the cells C having the maximum size (100 μm) or more from the cell suspension LA in the stage a 2. Therefore, in the second removal step, the cell filter 8 having a mesh opening size of 100 μm is used. Specifically, the cell filter 8 having a mesh size of 100 μm is inserted into the tube 85, and the cell suspension LA of stage a2 is stored in the pot 86.

Subsequently, the cell suspension LA was injected from the pot 86 into the tube 85 equipped with the cell filter 8(100 μm). Then, the cell suspension LA stored in the tube 85 after passing through the cell filter 8 is taken out, and the cell suspension LA of stage a3 is obtained. The cells C captured to the cell filter 8 are discarded. The cell suspension LA at stage A3 is a cell suspension including only the desired cells C to be transferred to the microtiter plate 4, and in the present embodiment, only the cells C having a size of 40 μm to 100 μm. The order of execution of the first and second removal steps is arbitrary, and the second removal step may be executed prior to the first removal step, in contrast to the example of fig. 8.

In the conventional pretreatment method for cell migration, the pretreatment is terminated at the stage of obtaining the cell suspension LA of the stage a 3. That is, the cell suspension LA in stage a3 is injected into the dispensing container 14. In contrast, in the present embodiment, the sorting step is further performed on the cell suspension LA of stage a 3. In the separation step, filtration is performed to separate the desired cells C contained in the cell suspension LA of stage a3 into two sizes of cells according to size. Here, an example of the cell C divided into 40 μm to 70 μm (first size) and 70 μm to 100 μm (second size) is shown.

In order to perform the sorting, in the sorting step, the cell filter 8 having a mesh opening size of 70 μm is used so that the cells C of the first size pass through but the cells C of the second size do not pass through. Specifically, the cell filter 8 having a mesh size of 70 μm is inserted into the tube 85, and the cell suspension LA of stage a3 is stored in the pot 86. Then, the cell suspension LA was poured from the pot 86 into the tube 85 equipped with the cell filter 8(70 μm).

Then, the cells C trapped in the cell filter 8 are taken out and dispersed in a cell culture medium or the like, thereby obtaining a cell suspension LA of stage a41 containing only cells C of a size of 70 μm to 100 μm. The cell suspension LA stored in the tube 85 through the cell filter 8 is removed to obtain a cell suspension LA in stage A42 containing only cells C of a size of 40 μm to 70 μm.

Thereafter, the following supply step is performed: the cells C classified into the cell suspension LA of stage a41 and the cell suspension LA of stage a42 are supplied to positions where they can be picked up by the tip 12 in the respective classified states. In the present embodiment, the supply step includes a step of injecting the cell suspensions LA in stages a41 and a42 into different dispensing containers 14, respectively, and a step of dispensing the cell suspensions LA from the dispensing containers 14 into different disks 2A and 2B, respectively.

The dispensing containers 14 into which the cell suspensions LA of the stages a41 and a42 have been injected are sequentially placed at the third placement position P3 of the base 1 or are juxtaposed at the same time. Then, the cell suspensions LA in stages a41 and a42 are individually aspirated from the respective dispensing containers 14 by the dispensing tip 13, moved to the upper space of the disks 2A and 2B (first placement position P1), and then dispensed. The dishes 2A, 2B can be separate dishes provided for different sorting vessels 11, or areas separated from each other in one dish 2 can be used as the dishes 2A, 2B.

[ advantages of performing the discriminating step ]

In the present embodiment, the above-described sorting step is further performed on the cell suspension LA of stage a 3. The advantage of performing this differentiation process is illustrated. FIG. 9 is a schematic diagram showing a cell picking operation with a pretreatment method of a comparative example. In the comparative example, the cell suspension LA in stage a3 is dispensed into the dispensing container 14. Therefore, cells C having a size of 40 μm to 100 μm are scattered in the dish member 2, and when picking up the cells by the tip 12, it is necessary to sort out the cells C to be moved from the cells C.

For example, consider the following case: from the cell suspension LA of stage A3, good cells C having a size of 80 μm. + -. 5 μm are picked up from the dish 2 and moved to the microtiter plate 4A, and good cells C having a size of 50 μm. + -. 5 μm are picked up from the dish 2 and moved to the microtiter plate 4B (the remainder is discarded). In this case, in the comparative example, it is necessary to perform a process of sorting high-quality cells C of 80 μm grade and high-quality cells C of 50 μm grade from the dish-shaped member 2 in which the cells C of a wide range of sizes of 40 μm to 100 μm are scattered.

One of the effective means for automatically sorting the cells C is machine learning. In this machine learning, for example, a large amount of sample data of an image of the cell C is analyzed to construct an algorithm for determining whether the cell C is a good cell. However, the criteria for sorting whether or not the cells are good may vary depending on the size of the cells C. For example, the sorting criteria are different between the 80 μm-sized cells C and the 50 μm-sized cells C described above. Therefore, if machine learning is performed on cells having a wide range of sizes, enormous sample data is required, and there arises a problem that it is difficult to construct an algorithm or a large amount of processing time is required. According to the comparative example, since it is necessary to perform analysis of sample data with cells C having a size of 40 μm to 100 μm as an object to create an algorithm as a sorting criterion, introduction of machine learning is accompanied by difficulty.

FIG. 10 is a view schematically showing a cell picking operation accompanied by the pretreatment method of the present embodiment. In the present embodiment, the cell suspension LA in stage a3 is separated into at least two types of cell suspensions LA according to size by the separation step. That is, the cell suspension LA in the stage A3 is divided into at least two types, that is, the cell suspension LA in the stage a41 containing a large-sized cell population and the cell suspension LA in the stage a42 containing a small-sized cell population, and the divided cell suspensions are supplied to the microtiter plates 4A and 4B via the dispensing container 14.

Therefore, the picking operation including the cell sorting operation by the cell transfer device S can be performed on the cell group in which the variation in cell size is suppressed. For example, when the cell suspension LA in stage A3 is subjected to a sorting step (filtration) by the cell filter 8 having a mesh opening size of 70 μm, the above-mentioned sorting operation of the cells C of 80 μm grade and the cells C of 50 μm grade is significantly facilitated. That is, it is sufficient to sort high-quality cells C of 80 μm grade from the dish 2A of the stage a41 cell suspension LA to which cells C of 70 to 100 μm are dispensed, and to sort high-quality cells C of 50 μm grade from the dish 2B of the stage a42 cell suspension LA to which cells C of 40 to 70 μm are dispensed. Therefore, when cell sorting is performed by introducing a machine learning method, since the variation in cell size is small, an accurate sorting criterion can be obtained with a relatively small number of sample data, and the processing can be simplified.

[ description of other embodiments ]

< modification of separation Process >

Fig. 8 shows an example in which the cell suspension LA at the stage A3 is classified into two types, i.e., the cell suspensions LA at the stages a41 and a42 in the classification step. In the separation step, the cell suspension LA in stage a3 may be separated into three or more types. FIG. 11 is a diagram showing a procedure of a pretreatment method in moving a cell according to a modification.

in the pretreatment method of fig. 11, the first and second removal steps are the same as in the example of fig. 8, but in the separation step, filtration is performed to separate the desired cells C contained in the cell suspension LA of the stage a3 into three types of cells according to size. Here, three examples of the cell C of 80 μm to 100 μm, the cell C of 60 μm to 80 μm, and the cell C of 40 μm to 60 μm are shown. In this case, two kinds of cell filters having mesh openings of 60 μm and 80 μm were used as the cell filter 8.

To illustrate a specific example of the sorting operation, first, the cell filter 8 having a mesh size of 80 μm is fitted into the tube 85, and the cell suspension LA in stage a3 is stored in the pot 86. Then, the cell suspension LA was poured from the pot 86 into the tube 85 equipped with the cell filter 8(80 μm). Then, the cells C trapped in the cell filter 8 are taken out and dispersed in a cell culture medium or the like, thereby obtaining a cell suspension LA of stage a41 containing only cells C of a size of 80 μm to 100 μm.

Then, the cell suspension LA stored in the tube 85 through the cell filter 8 is taken out to obtain a cell suspension LA containing only cells C of a size of 40 μm to 80 μm. Next, the cell filter 8 having a mesh size of 60 μm is inserted into the tube 85, and the cell suspension LA containing the cells C having a size of 40 μm to 80 μm is stored in the pot 86. The cell suspension LA is injected from a pot 86 into a tube 85 equipped with a cell filter 8(60 μm).

Then, the cells C trapped in the cell filter 8 are taken out and dispersed in a cell culture medium or the like, thereby obtaining a cell suspension LA of stage a42 containing only cells C having a size of 60 μm to 80 μm. The cell suspension LA stored in the tube 85 through the cell filter 8 is removed to obtain a cell suspension LA in stage A43 containing only cells C of a size of 40 μm to 60 μm. The cell suspensions LA obtained in the stages a41, a42, and a43 were dispensed to the respective disks 2A, 2B, and 2C.

< example of use for mesh size control >

The cell moving apparatus S of the present embodiment includes an input unit 55 (fig. 5) that receives input of various data from a user. The input unit 55 may receive an input of the mesh size of the cell filter 8 used in the sorting step, and may use the input as a criterion for sorting the cells C in the sorting unit 76.

If the previous example of FIG. 8 is used, the cell suspension LA in stage A41 is obtained by using two cell filters 8 having mesh sizes of 70 μm and 100 μm. In this case, the user inputs data of 70 μm or 100 μm screen size to the input unit 55. The sorting unit 76 automatically sets the cell size (diameter) to 70 μm to 100 μm as one of the criteria for sorting the cells C. The sorting unit 76 reads out other sorting criteria (such as an algorithm based on machine learning) set for the cells C having the size of 70 μm to 100 μm from the storage unit 77. Then, if the cell suspension LA of the stage a41 is dispensed into the dish 2A, the sorting unit 76 applies the above sorting criterion to each image of the cells C placed on the dish 2A captured by the camera unit 5 to determine the quality of each cell C. This makes it possible to improve the efficiency of the cell migration operation.

In addition to the above, the following may be provided: the input unit 55 also receives an input of at least one of the upper limit and the lower limit of the size of the cell C to be moved, and the sorting unit 76 processes the input upper limit or the input lower limit of the size of the cell C as the element of the sorting criterion. Here, it is desirable that the sorting unit 76 does not receive input of at least one of the upper limit and the lower limit of the size when the upper limit or the lower limit of the size input to the input unit 55 is inconsistent with the mesh size of the cell filter 8 input before.

A specific example will be described with reference to fig. 12. FIG. 12 is a diagram showing a preferred example of the operation of picking up cells from the pretreatment at the time of cell transfer. Similarly to the example of fig. 8, it is assumed that the cell suspension LA of stage A3 is obtained using the cell filter 8 having a mesh size of 40 μm and 100 μm in the first and second removal steps, and the cell suspension LA of stages a41 and a42 is obtained using the cell filter 8 having a mesh size of 70 μm in the separation step. Then, the cell suspension LA of stage a41 is dispensed into the dish 2 of the sorting container 11. Here, it is assumed that the size of the cells C to be transferred to the microtiter plate 4, that is, the cells C to be picked up is 80 μm to 90 μm.

In this case, as in the above example, the input unit 55 receives input of data indicating that the mesh size of the cell filter 8 used to obtain the cell suspension LA in the stage a41 is 70 μm or 100 μm from the user. Further, the input unit 55 receives input of data indicating that the size of the cell C to be picked up is 80 μm to 90 μm. Then, the sorting unit 76 automatically sets the condition that the cell size is 80 μm to 90 μm as one of the sorting criteria for the cells C, and reads out the other sorting criteria set for the cells C of the size of 80 μm to 90 μm from the storage unit 77 to set the designated selection criteria.

Here, the following may occur: the user performs input that contradicts the mesh size (70 μm, 100 μm) previously input for the cell size of the pickup object. For example, the user inputs the upper limit of the cell size of the pickup object to the input unit 55, for example, 50 μm or the lower limit thereof to 40 μm. In this case, the sorting unit 76 prompts the user to re-input the input by, for example, displaying an error message on the display unit 54 without accepting the input.

if the mesh size and the cell size of the object to be picked are inputted to the input section 55 without contradiction, the sorting section 76 performs the quality determination of the cells C based on the sorting criterion corresponding to the cell size (80 μm to 90 μm) of the object to be picked. Then, the cells C sorted as the moving object are aspirated from the sorting container 11 through the tip 12 and moved to the microtiter plate 4.

< modification of dish Member and microtiter plate >

FIG. 13 is a diagram showing another preferred example of the operation of picking up cells from the pretreatment during cell transfer. Here, the following example is shown: a plurality of dishes 2A to 2D to which cells C are to be dispensed are prepared, and wells 41 are assigned to each dish 2A to 2D in the microtiter plate 4.

4 disks 2A to 2D are installed in the sorting vessel 11. These dishes 2A to 2D are immersed in the medium L (see fig. 1) in the sorting vessel 11, but are isolated from each other by partition walls or the like. The disks 2A to 2D are dispensed with cells C of different sizes. That is, cell suspensions LA obtained by applying cell filters 8A, 8B, 8C, and 8D having different mesh sizes are dispensed to the partitions in which the disks 2A, 2B, 2C, and 2D are arranged, respectively.

The plurality of wells 41 provided in the microtiter plate 4 are determined in advance as destinations to which the cells C picked up from the disks 2A to 2D are to be transferred. That is, the cells C picked up from the dish 2A are distributed in advance such that the cells C are moved to the wells 41 in the first row of the microtiter plate 4 and the cells C picked up from the dish 2B are moved to the wells 41 in the second row.

then, the cells C picked up from each dish 2A to 2D by the tip 12 are discharged into the holes 41 respectively assigned to the dishes 2A to 2D subjected to picking out among the plurality of holes 41. This allows the desired cells C to move to the holes 41 in a state where their sizes are distinguished or in a state where a difference in the sizes of the cells C can be recognized. Therefore, various processing operations in the subsequent microtiter plate 4 can be appropriately performed according to the cell size.

The specific embodiments described above mainly include the invention having the following configurations.

a pretreatment method for cell movement according to an aspect of the present invention is a pretreatment method for picking up a desired cell from an aggregate including a plurality of cells of different sizes using a cell moving apparatus including a cell pickup unit, and moving the cell to a predetermined destination, the pretreatment method including: a distinguishing step of distinguishing the desired cell among the plurality of cells contained in the aggregate into at least two types of cells according to size; and a supplying step of supplying the cells subjected to the differentiation to a position where the cells can be picked up by the pickup unit in a state where the cells are differentiated from each other.

According to the pretreatment method, the desired cells are divided into at least two cells according to size. Thus, the desired cells are distinguished into at least two of a cell population having a large size and a cell population having a small size. Then, the desired cells are supplied to the pickup position in a state where they are distinguished from each other. Therefore, it is possible to perform a picking operation including a cell sorting operation by the cell transfer device described later on the cell group in the divided state. That is, since the picking operation can be performed on the cell group in which the size variation of the cells is suppressed, the working time can be shortened.

In the pretreatment method, preferably, the desired cells are cells having a size exceeding a predetermined minimum size, the pretreatment method further comprising: a first removal step of removing cells below the minimum size from the pool.

Further, preferably, the desired cells are cells having a size smaller than a predetermined maximum size, and the pretreatment method further comprises: a second removal step of removing cells having the maximum size or larger from the aggregate.

according to these pretreatment methods, the oversized cells and the undersized cells can be removed from the aggregate by the first and second removal steps. Thereby, only the desired cells can be extracted from the aggregate. Therefore, by performing the distinguishing step, a plurality of cells within a desired size range can be more reliably distinguished into at least two kinds of cells.

In the pretreatment method, it is preferable that the aggregate is a cell suspension in which a plurality of cells having different sizes are dispersed, and in the separation step, a filter is used which allows cells having a first size to pass therethrough but does not allow cells having a second size larger than the first size to pass therethrough.

according to this pretreatment method, the differentiating operation corresponding to the size of the desired cells in the differentiating step can be efficiently performed by the filter.

In the pretreatment method, it is preferable that, in the second removal step, a filter having a mesh size of 100 μm is used, and in the discrimination step, a filter having a mesh size of 70 μm is used.

As a filter for cell filtration, a filter having a mesh size of 70 μm or 100 μm is available as an inexpensive multipurpose product. By using such a filter, it is possible to remove cells of a size of 100 μm or more from the cells contained in the aggregate in the second removal step, and to distinguish a cell population of less than 100 μm into two types of cells of a size of less than 70 μm and a size between 70 μm and 100 μm.

In the pretreatment method, it is preferable that in the first removal step, a filter having a mesh size of 40 μm is used, in the second removal step, a filter having a mesh size of 100 μm is used, and in the classification step, a filter having a mesh size of 70 μm is used.

As a filter for cell filtration, in addition to the above-mentioned mesh size, a filter having a mesh size of 40 μm is available as an inexpensive multipurpose product. By using such a filter, cells having a size of not more than 40 μm can be removed from the cells contained in the aggregate in the first removal step, and cells having a size of 100 μm or more can be removed in the second removal step. Further at the differentiating step, differentiating the cell population of 40 μm to 100 μm into two cells between a size of 40 μm to 70 μm and between a size of 70 μm to 100 μm.

A cell transfer apparatus according to another aspect of the present invention is a cell transfer apparatus used after applying the above-described pretreatment method for cell transfer, including: a pick-up unit for the cells; a moving unit that moves the pickup unit in a predetermined direction; a sorting unit that sorts the cells to be moved out of the cells supplied in the supplying step; and an input unit that receives an input of a mesh size of the filter to be used; wherein the sorting unit performs the sorting based on a sorting criterion predetermined for cells having a size corresponding to the input mesh size, the picking unit picks up the cells sorted by the sorting unit, and the moving unit moves the picking unit that has picked up the cells to a predetermined destination.

According to the cell transfer device, the sorting unit performs the sorting based on a sorting criterion predetermined for the cell having the size corresponding to the inputted mesh size. Here, the size of the cells is already substantially uniform based on the execution of the sorting step, and therefore, the sorting criterion does not need to be targeted for cells of a wide range of sizes. For example, when cell sorting is performed by introducing a machine learning method, since the variation in cell size is small, an accurate sorting criterion can be obtained with a relatively small number of sample data, and the processing can be simplified.

In the cell transfer device, it is preferable that the input unit further receives an input of at least one of an upper limit and a lower limit of a size of the cell to be transferred, the sorting unit processes the upper limit or the lower limit of the size as the element of the sorting criterion, and the input unit does not receive an input of at least one of the upper limit and the lower limit of the size when the upper limit or the lower limit of the size is inconsistent with the mesh size.

According to this cell transfer device, when there is the conflict, the input unit does not accept input of the upper limit or the lower limit of the size. Therefore, in addition to the limitation of the variation in cell size based on the size of the mesh to be input, the variation in cell size of the cells to be moved is also limited.

In the cell transfer device, it is preferable that, in the supplying step, a plurality of dishes capable of holding cells are prepared, the cells are held by the dishes for the respective cells to be distinguished, a microtiter plate having a plurality of wells for receiving the cells is arranged at the specified transfer destination, and the picking unit includes a tip for performing suction and discharge of the cells, and the cells are discharged from the tip to the wells to which the plurality of wells are allocated after being sucked by the tip for each dish and transferred to the microtiter plate.

according to this cell transfer device, the desired cells can be transferred to the wells of the microtiter plate while being distinguished in their sizes or while being able to recognize a difference in the sizes of the cells. Therefore, the subsequent processing operation in the microtiter plate can be appropriately performed according to the size of the cells.

According to the present invention described above, it is possible to provide a pretreatment method capable of efficiently moving a desired cell using a cell transfer device provided with a cell pickup unit, and a cell transfer device to which the method is applied.