JP2020068682A - Cell-conveying system - Google Patents

Abstract

To provide a cell-conveying system that can obtain a clear image not only when sucking cells into a first container but also when discharging cells into a second container.SOLUTION: A cell-conveying system (1) comprises: a needle (33); a first microscope (microscope 11) which observes an area near a tip (332) of the needle (33) in the first container (schale SH); and a second microscope (microscope 13) which observes an area near the tip (332) in a second container (PCR tube Pij).SELECTED DRAWING: Figure 1

Description

  The present invention relates to a cell delivery system that delivers cells from a first container to a second container.

  For cell analysis, from a Petri dish (an example of the first container) in which the cell population is cultured, a plurality of PCR tubes (for example, 96 wells and 384 wells) constituting the PCR plate (of the second container). In one example, it is necessary to isolate and dispense a specified number of cells (for example, one cell at a time).

  In order to analyze a single cell of interest from the cell population in culture in a petri dish, observe the inside of the petri dish under a highly functional and highly accurate optical microscope and aspirate the single cell of interest with a glass needle or the like. Therefore, there is a need for quick picking and single ejection of picked single cells into PCR tubes. Here, in order to confirm whether or not cells can be actually ejected one by one, it is necessary to observe the inside of the PCR tube even when the cells are ejected into the PCR tube. In the following, an optical microscope is simply referred to as a microscope.

  For example, in the cell delivery system described in Patent Document 1, as shown in FIG. 9, observation of a cell population in a petri dish and observation during picking of cells, and observation during ejection in a PCR tube are performed. Is carried out using one microscope.

JP-A-2018-68169 (Published May 10, 2018)

  However, when observing the inside of the petri dish and the inside of the PCR tube with one microscope, cells existing in the first container and the second container having different materials and shapes are used with the same one microscope. Observe. Therefore, the conventional cell transport system described in Patent Document 1, for example, has a problem that it is difficult to obtain a clear image during ejection.

  A cell delivery system according to one aspect of the present invention is made in view of the above-mentioned problems, and an object thereof is a cell delivery system for delivering cells from a first container to a second container. It is an object of the present invention to provide a cell transport system that can obtain a clear image when cells are ejected in the second container in addition to when cells are picked in the container.

  In order to solve the above problems, a cell delivery system according to an aspect of the present invention sucks one or a plurality of cells from a first container that houses a plurality of cells, A needle for discharging each into one or a plurality of second containers, and a first microscope for observing the vicinity of the tip when the tip of the needle is located in the first container, A second microscope for observing the vicinity of the tip when the tip is located in the second container or above the opening of the second container.

  Further, in order to solve the above problems, a cell delivery system according to an aspect of the present invention sucks one or a plurality of cells from a first container that houses a plurality of cells, and A needle for ejecting each of the plurality of cells into each of the one or more second containers; and a first part for observing the vicinity of the tip when the tip of the needle is located in the first container And a second microscope for observing the vicinity of the tip when the tip is located in the second container.

  A cell delivery system according to one aspect of the present invention is a cell delivery system for delivering cells from a first container to a second container, and in addition to the case of aspirating cells in the first container, a second It is possible to provide a cell transport system capable of obtaining a clear image even when cells are discharged in the container.

(A) is a cross-sectional arrow view showing an outline of the cell transport system according to the first embodiment of the present invention. (B) is an enlarged cross-sectional view in which the vicinity of the tip of a needle included in the cell transport system shown in (a) is enlarged. (C) is a plan view of a PCR plate used in the cell transfer system according to the first embodiment. It is a block diagram of the cell delivery system shown to (a) of FIG. It is a perspective view of the 2nd microscope and optical stage with which the cell conveyance system shown to Fig.1 (a) is equipped. It is a cross-sectional arrow view which shows the outline of the cell delivery system which concerns on the 2nd Embodiment of this invention.

First Embodiment
The cell delivery system 1 according to the first embodiment of the present invention will be described with reference to FIGS. 1 to 3. FIG. 1A is a cross-sectional arrow view showing the outline of the cell transport system 1. FIG. 1B is an enlarged cross-sectional view in which the vicinity of the tip 332 of the needle 33 included in the cell transport system 1 is enlarged. FIG. 1C is a plan view of the PCR plate PP used in the cell transport system 1. FIG. 2 is a block diagram of the cell transport system 1. FIG. 3 is a perspective view of the microscope 13 and the optical stage 43 included in the cell transfer system 1.

<Platelet SH and PCR tube Pij>
The cell transfer system 1 is a cell transfer system that transfers cells from the Petri dish SH to the PCR tube Pij.

  The petri dish SH, which is an example of the first container described in the claims, is made of glass (transparent in the present embodiment) that is transparent to the observation light L1 described later. A plurality of cells (not shown in FIG. 1) are housed in the petri dish SH together with the culture medium CF1.

  The PCR plate PP, which is an example of the second container described in the claims, is made of a resin (transparent in the present embodiment) that is transparent to the observation light L2 described later, and a plurality of them are used. The PCR tube Pij, which is the concave portion, is a resin plate formed in a matrix of m rows and n columns (see (c) of FIG. 1). The PCR plate PP is sometimes called a well plate. The PCR plate PP (well plate) may be made of translucent (more preferably transparent) glass instead of resin.

  In the present embodiment, a PCR plate PP having a total of 96 PCR tubes Pij consisting of 12 rows and 8 columns is used. That is, i is an integer of 1 or more and 12 or less, and j is an integer of 1 or more and 8 or less. It should be noted that FIG. 1A shows a state in which the tip 332 of the needle 33 is positioned inside the PCR tube P33 among the PCR tubes P31 to P38 in the row where m = 3.

  In the PCR plate PP, the pitch Py, which is the interval between adjacent rows, is common between all rows, and the pitch Px, which is the interval between adjacent columns is common between all columns. The pitch Py and the pitch Px are common (see (c) of FIG. 1).

  However, the number of PCR tubes Pij included in the PCR plate PP is not limited. For example, an example of the second container may be a PCR plate PP having a total of 384 PCR tubes Pij each having 12 rows and 32 columns. Each PCR tube Pij is filled with the culture medium CF2 (see (a) of FIG. 1).

  Each of the dish SH, which is an example of the first container, and the PCR tube Pij, which is an example of the second container, are not constituent elements of the cell transport system 1. As a matter of course, each of the first container and the second container can be appropriately selected depending on the application (for example, cell analysis).

<Configuration of cell transport system 1>
The cell transfer system 1 is shown in FIG. 1 with the microscopes 11 and 13, the light sources 16a, 16b, 17a, 17b and 17c, the stages 21 and 22, and the cell transfer robot 30 shown in FIG. The optical stage 43 and an anti-vibration table not shown in FIGS. 1 and 3 are provided.

(Microscope 11)
A microscope 11, which is an example of a first microscope described in the claims, is placed on a vibration isolation table. As shown in FIG. 2, the microscope 11 includes a lens group 11a composed of a plurality of lenses and an image sensor 11b. Some of the lenses included in the lens group 11a form the objective lens 12 shown in FIG.

  The image sensor 11b, which is an example of the first image sensor described in the claims, captures an image formed by the lens group 11a and generates first image data representing the image.

  The microscope 11 is a microscope having specifications suitable for detailed observation and analysis of cells, and has a higher magnification and a higher function than a microscope 13 described later. Therefore, as compared with the microscope 13, the microscope 11 has a large numerical aperture and a short working distance (see the observation light L1 shown in FIG. 1A). The optical axis AL1 of the microscope 11 is an example of the first optical axis described in the claims.

  The stage 21 that constitutes a part of the microscope 11 is a stage for mounting the dish SH. The stage 21 includes an outer frame 21a, a plate 21b, and a stage mechanism 21c.

  The plate 21b is a plate-shaped member on which the Petri dish SH is placed, and is made of glass having translucency for the observation light L1.

  The outer frame 21a is a frame-shaped member for supporting the plate 21b, and is shaped to surround the outer edge of the plate 21b.

  The outer frame 21a is supported by a stage mechanism 21c schematically shown in FIG. The position of the outer frame 21a can be adjusted by using the stage mechanism 21c.

  In the present embodiment, the stage mechanism 21c can translate the outer frame 21a in each of the x-axis direction, the y-axis direction, and the z-axis direction, and the main surface of the outer frame 21a (the main surface of the plate 21b). Can be tilted with respect to the xy plane.

  In this embodiment, the stage mechanism 21c is configured so that the position of the outer frame 21a can be adjusted by either a manual method or an electric method. The stage mechanism 21c includes a plurality of motors in order to electrically drive the stage mechanism 21c. When the stage mechanism 21c is driven electrically, the stage mechanism 21c is controlled by the first stage controller 10d included in the controller 10 of the cell transfer system 1. The mode of the plurality of motors is not limited, and may be, for example, a stepping motor or a servo motor.

  Since the stage 21 is configured as described above, in a state where the microscope 11 is adjusted so that the tip 332 of the needle 33, which will be described later, is in focus, the relative relationship between the tip 332 and the microscope 11 is not changed ( That is, only the position of the petri dish SH can be adjusted while the focus is maintained. Therefore, the operator can easily aspirate desired cells using the cell transfer robot 30 described later.

(Microscope 13)
As shown in FIG. 3, a microscope 13, which is an example of a second microscope described in the claims, is fixed to a vibration isolation table via a spindle 41, an arm 42, and an optical stage 43. ..

  As shown in FIG. 2, the microscope 13 includes a lens group 13a composed of a plurality of lenses and an image sensor 13b. The microscope 13 is a zoom lens in which the magnification of the image obtained by rotating the zoom ring 15 provided in the cylindrical casing 14 is variable.

  In the cell transfer system 1, a zoom lens is adopted as the microscope 13, but a microscope equipped with a plurality of objective lenses having different magnifications and a revolver for switching these plurality of objective lenses is adopted as the microscope 13. You may.

  The microscope 13 is a microscope having specifications suitable for observing the state of ejecting cells into the PCR tube Pij from the tip 332 of the needle 33 described later, and has a lower magnification than the microscope 11. It is lightweight and compact.

  By adopting the zoom lens as the microscope 13, the magnification can be changed without exchanging a part (for example, an objective lens) of the lenses constituting the microscope or rotating the revolver without interrupting the observation. be able to. That is, it is possible to perform observation with the entire area of the opening AP pulled or to observe the vicinity of the tip 332 of the needle 33 in a zoomed state without performing an operation that interrupts the observation. Therefore, the microscope 13 has a smaller numerical aperture and a longer working distance than the microscope 11 (see the observation light L2 shown in FIG. 1A). The optical axis AL2 of the microscope 13 is an example of the second optical axis described in the claims.

  The main shaft 41 is a columnar member made of metal (stainless steel in this embodiment). The lower end of the main shaft 41 is fixed to the surface of the vibration isolation table so that the central axis A41 of the main shaft 41 is perpendicular to the surface of the vibration isolation table.

  An optical stage 43, which will be described later, is an optical stage that is fixed to the upper end of the main shaft 41 and has a rectangular parallelepiped outer shape. The central axis A42 of the arm 42 is perpendicular to one of the side surfaces forming the optical stage 43, in other words, the central axis A42 of the arm 42 is parallel to the surface of the vibration isolation table. As described above, one end of the arm 42 is fixed to one side surface of the optical stage 43.

  The image pickup device 13b is fixed to the other end of the arm 42 so that the optical axis AL2 is perpendicular to the surface of the vibration isolation table.

  The image sensor 13b, which is an example of the second image sensor described in the claims, captures an image formed by the lens group 13a and generates second image data representing the image.

  The stage 22 that constitutes a part of the microscope 13 is a stage for mounting the PCR plate PP, and is an example of the second adjusting mechanism described in the claims. The stage 22 includes an outer frame 22a, a plate 22b, and a stage mechanism 22c.

  The stage 22 is configured similarly to the stage 21. That is, the outer frame 22a of the stage 22 is configured similarly to the outer frame 21a of the stage 21, the plate 22b of the stage 22 is configured similarly to the plate 21b of the stage 21, and the outer frame 22a is 1A is supported by a stage mechanism 22c schematically shown in FIG. Therefore, a detailed description of the stage 22 is omitted here.

  Since the stage 22 is configured in the same manner as the stage 21, without changing the relative relationship between the tip 332 and the microscope 13 in a state where the microscope 13 is adjusted so that the tip 332 of the needle 33 described later is in focus. Only the position of the PCR tube Pij can be adjusted (that is, in the focused state). Therefore, the operator can easily determine the position where the cells are discharged by using the cell transfer robot 30 described later.

  It should be noted that FIG. 1A illustrates a case where cells are ejected with the tip 332 positioned inside the PCR tube Pij. However, the position of the tip 332 when discharging cells is not limited to the position inside the PCR tube Pij, and may be above the opening AP of the PCR tube Pij. When the cells are discharged from above the opening AP, the position of the cells after being discharged from the tip 332 in the z-axis direction is significantly lower than that before being discharged from the tip 332. Therefore, it becomes difficult to capture the cells as a clear image before and after the ejection. However, since the operator can check the cells discharged from the tip 332, it is possible to check that one cell has been discharged even when the cells are discharged from above the opening AP.

  It should be noted that, in that the cells can be continuously captured as a clear image before and after the ejection, when the cells are ejected with the tip 332 positioned in the PCR tube Pij, the tip is located above the opening AP. It is preferable to discharge cells in a state where 332 is positioned.

(Optical stage 43)
The optical stage 43 shown in FIG. 3 is an example of a first adjustment mechanism that adjusts the positions of the lens group 13a and the image pickup device 13b of the microscope 13.

  The optical stage 43 allows the arm 42 to translate in the directions of arrow A3 parallel to the x-axis direction, the direction of arrow A4 parallel to the y-axis direction, and the direction of arrow A5 parallel to the z-axis direction. It is configured. Therefore, by adjusting the optical stage 43, when the tip 332 is located inside the PCR tube Pij, the cell transport system 1 focuses the observation light L2 in the x-axis direction, the y-axis direction, and the z-axis direction. Can be moved.

  Further, the optical stage 43 is configured such that the arm 42 can be rotated in the direction around the central axis A42 of the arm 42 (direction of arrow A6). Therefore, by adjusting the optical stage 43, the cell transport system 1 can incline the optical axis AL2 along the yz plane when the tip 332 is located in the PCR tube Pij. In other words, the cell transport system 1 can change the depression angle when the PCR tube Pij is viewed from the image sensor 13b.

Further, the optical stage 43 is configured to be able to change the angle θ ₄₂ (elevation angle or depression angle) of the central axis A42 with respect to the surface of the vibration isolation table (see arrow A7). Therefore, by adjusting the optical stage 43, the cell transport system 1 can incline the optical axis AL2 along the zx plane when the tip 332 is located in the PCR tube Pij. In other words, the cell transport system 1 can change the depression angle when the PCR tube Pij is viewed from the image sensor 13b.

  In this embodiment, the optical stage 43 is configured so that the position of the arm 42 can be adjusted by either a manual method or an electric method. The optical stage 43 is equipped with a plurality of motors in order to drive the optical stage 43 electrically. When the optical stage 43 is driven electrically, the optical stage 43 is controlled by the optical stage controller 10b included in the controller 10 of the cell transport system 1. The mode of the plurality of motors is not limited, and may be, for example, a stepping motor or a servo motor.

  Further, in the present embodiment, the optical stage 43 collectively adjusts the positions of the lens group 13a, the image pickup device 13b, the housing 14, and the zoom ring 15 that configure the microscope 13. However, the optical stage 43 may be configured to adjust the position of at least a part of the microscope 13 (for example, one of the lens group 13a and the image sensor 13b). In that case, although the lens group 13a and the image pickup device 13b are optically coupled, they may be physically configured so that their positions can be adjusted separately.

(Light sources 16a, 16b, 17a, 17b, 17c)
The light sources 16a and 16b are light sources that form a part of the microscope 11 and that illuminate the dish SH with illumination light (see (a) of FIG. 1). Similarly, the light sources 17a, 17b, and 17c are light sources that form a part of the microscope 13 and that illuminate the PCR tube Pij with illumination light.

  The light source 17a is a light source that illuminates a PCR tube Pij into which a tip 332 of a needle 33, which will be described later, is inserted, and is arranged below the stage 22 and on the optical axis AL2. The light source 17b is a light source that obliquely illuminates the PCR tube Pij into which the tip 332 is inserted, and the depression angle when viewed from below the stage 22 and from the PCR tube Pij into which the needle is inserted is a predetermined angle (eg, 45 degrees). The light source 17c is a light source that laterally illuminates the PCR tube Pij into which the tip 332 is inserted, and has a predetermined elevation angle when viewed from above the stage 22 and from the PCR tube Pij into which the tip 332 is inserted. For example, it is arranged at a position of 25 degrees.

  The light source 17a is fixed to, for example, a vibration isolation table via a mount capable of adjusting the position of the light source 17a. Here, the position of the light source 17a includes the coordinates on the surface of the vibration isolation table (in the xy plane) and the direction of the illumination light. That is, the mount which is the fourth adjusting mechanism described in the claims is configured so that at least one of the coordinates of the light source 17a in the xy plane and the direction of the illumination light of the light source 17a can be adjusted. Has been done. The mount can be configured by, for example, a magnet base that can be fixed to the surface of the vibration isolation table, and a hinge that is fixed on the magnet base.

  Like the light source 17a, each of the light source 17b and the light source 17c is fixed to the vibration isolation table via a mount whose position can be adjusted.

  In addition, each of the light sources 17a, 17b, and 17c is configured to be able to control whether or not to individually light. The method for selecting the light source to be turned on from the light sources 17a, 17b, 17c is not limited. In the cell delivery system 1 of the present embodiment, the second selection unit 10h included in the control unit 10 selects the light source to be turned on. Alternatively, a switch may be provided for each of the light sources 17a, 17b, and 17c, and the operator may operate the switches to select the light source to be turned on. The control unit 10 will be described later.

  The light source 16a has the same configuration as the light source 17a except that the light source 16a is arranged above the stage 21 as the microscope 11 is arranged below the stage 21. The light source 16b has the same structure as the light source 17c. Therefore, a detailed description of the light sources 16a and 16b is omitted here.

  In addition, in the present embodiment, the microscope 11 includes two light sources 16a and 16b, and the microscope 13 includes three light sources 17a, 17b and 17c. However, the number of light sources and the positions of the light sources provided in each of the microscope 11 and the microscope 13 are not limited and can be appropriately determined. Further, the wavelength of the illumination light emitted by each of the light sources 16a, 16b, 17a, 17b, 17c can be appropriately determined. In particular, when performing fluorescence observation with the microscope 11, the wavelength of the illumination light emitted by any of the light sources 16a and 16b may be included in the visible light region or may be included in the ultraviolet light region. ..

(Cell transfer robot 30)
As shown in FIG. 1A, the cell transfer robot 30 includes a main shaft 31, an arm 32, and a needle 33. In addition, the cell transfer robot 30 further includes a pump not shown in FIG.

  The main shaft 31 is a columnar member made of metal (stainless steel in this embodiment). The lower end of the main shaft 31 is fixed to the surface of the vibration isolation table so as to be perpendicular to the surface of the vibration isolation table so that the center axis A31 of the main shaft 31 is rotatable about the axis of the center axis A31. There is. Therefore, the main shaft 31 can rotate in the direction of the arrow A2 shown in FIG.

  The arm 32 is a robot arm including a plurality of links and one or a plurality of joints (a plurality of joints in this embodiment). In FIG. 1A, among the plurality of links and the plurality of joints, only the link 32a that constitutes one end and the link 32b that constitutes the other end are shown, and the intermediate portion of the arm 32 is shown. Illustrations of other links and a plurality of joints configuring the above are omitted. When the arm 32 is compared to a human arm, the link corresponds to the bone and the joint corresponds to the joint.

  The link 32 a, which is one end of the arm 32, is fixed to the upper end of the main shaft 31. The link 32b that is the other end of the arm 32 is supported by a plurality of links and a plurality of joints. The position of the link 32b can be arbitrarily controlled by controlling a plurality of joints. Therefore, the position of the link 32b (in other words, the position of the tip 332 of the needle 33 described later) can be adjusted along each of the x-axis direction, the y-axis direction, and the z-axis direction.

  In particular, since the position of the tip 332 can be moved up and down in the direction of arrow A1 along the z-axis direction, (1) the tip 332 can be inserted into the dish SH without moving each of the stage 21 and the stage 22. Alternatively, the petri dish SH can be retracted from the dish SH, and (2) the tip 332 can be inserted into the PCR tube Pij or retracted from the PCR tube Pij.

  In the present embodiment, the arm 32 is configured so that the position of the link 32b can be adjusted by both manual and electric methods. The arm 32 includes a plurality of motors in order to electrically drive the arm 32. When the arm 32 is driven electrically, the arm 32 is controlled by the arm control unit 10a included in the control unit 10 of the cell transfer system 1. The mode of the plurality of motors is not limited, and may be, for example, a stepping motor or a servo motor.

  The link 32b fixes one end of the needle 33, which is a tubular member whose both ends are open, and connects the pipe connected to the pump in an airtight state to the one end (the pump and the pump). The piping connected to the pump is not shown in FIG. Although detailed description of the pump is omitted in the present embodiment, the pump is configured to be capable of sucking and discharging a minute volume of fluid (gas or liquid). Since the pump and the needle 33 are connected in an airtight state, the needle 33 can suck or discharge the culture solution CF1 or the culture solution CF2 in accordance with the suction or discharge of the fluid performed by the pump.

  The needle 33 includes a main portion 331, a tip 332, and a tapered portion 333 (see (a) and (b) of FIG. 1). The needle 33 is made of glass that is transparent to the observation light L1 and the observation light L2 (transparent in the present embodiment). Note that the material forming the needle 33 is not limited to glass, and may be any material (for example, resin) that is transparent to the observation lights L1 and L2.

  The main portion 331 includes one end of the needle 33. The main portion 331 is configured such that the inner diameter and the outer diameter are substantially constant over the entire section. The main portion 331 is bent in the middle section. The bent angle of the main portion 331 may be an angle that allows the tip 332 to be inserted into the PCR tube Pij in a state where the needle 33 and the inner wall of the PCR tube Pij do not interfere with each other.

  The tip 332 constitutes the other end of the needle 33. The tip 332 is configured such that the inner diameter and the outer diameter are smaller than the inner diameter and the outer diameter of the main portion 331 over the entire section.

The tapered portion 333 smoothly bends the main portion 331 and the tip 332, and is bent so that the central axis A331 of the main portion 331 and the central axis A332 of the tip 332 form an angle θ ₃₃ . Since the central axis A331 and the central axis A332 form an angle θ ₃₃ , the tip 332 can be inserted into the PCR tube Pij without interference between the needle 33 and the inner wall of the PCR tube Pij, and the microscope 11 Can be used to observe the longitudinal cross section of the tip 332. Therefore, the displacement in the z-axis direction of the cells can be suppressed before and after the aspiration of the cells in the dish SH and before and after the ejection of the cells in the PCR tube Pij. Cell vividness can be obtained over the entire range. The angle θ ₃₃ can be appropriately determined so that the angle formed by the central axis A 332 and the xy plane becomes small according to the angle at which the main portion 331 is bent.

(Control unit 10)
As shown in FIG. 2, the control unit 10 of the cell transfer system 1 includes an arm control unit 10a, an optical stage control unit 10b, a first selection unit 10c, a first stage control unit 10d, and a first stage control unit 10d. An image data acquisition unit 10e, a first determination unit 10f, a first flag addition unit 10g, a second selection unit 10h, a second stage control unit 10i, and a second image data acquisition unit 10j. , And a second determination unit 10k and a second flag addition unit 10l.

  The arm controller 10 a controls the arm 32 to adjust the position of the tip 332.

  By controlling the optical stage 43, the optical stage controller 10b collectively adjusts the positions of the lens group 13a, the image sensor 13b, the housing 14, and the zoom ring 15 that configure the microscope 13.

  The first selection unit 10c selects a light source to be turned on from the light sources 16a and 16b.

  The first stage control unit 10d controls the stage mechanism 21c of the stage 21 to adjust the positions of the outer frame 21a and the plate 21b, that is, the petri dish SH.

  The lens group 11a of the microscope 11 forms an image of a nearby region including the tip 332 on the light receiving surface of the image pickup element 11b when the tip 332 is located inside the dish SH.

  The image pickup device 11b generates first image data representing an image of a near area including the tip 332 formed by the lens group 11a.

  The first image data acquisition unit 10e acquires the first image data from the image sensor 11b.

  The first determination unit 10f refers to the image of the vicinity region including the tip 332 in the petri dish SH, which is represented by the first image data, and determines whether or not one cell is sucked from the tip 332 into the needle 33. To judge.

  When the determination result by the first determination unit 10f is negative, the first flag addition unit 10g adds a flag indicating that one cell is not aspirated to the first image data. ..

  The second selection unit 10h selects the light source to be turned on from the light sources 17a, 17b, 17c.

  The second stage controller 10i adjusts the positions of the outer frame 22a and the plate 22b, that is, the positions of the PCR tubes Pij by controlling the stage mechanism 22c of the stage 22.

  When the tip 332 is located inside the PCR tube Pij, the lens group 13a of the microscope 13 forms an image of a nearby area including the tip 332 on the light receiving surface of the image sensor 13b.

  The image pickup device 13b generates second image data representing an image of a near area including the tip 332 formed by the lens group 13a.

  The second image data acquisition unit 10j acquires the second image data from the image sensor 13b.

  The second determination unit 10k refers to the image of the vicinity region including the tip 332 in the PCR tube Pij represented by the second image data, and refers to one cell discharged from the tip 332 into the PCR tube Pij. Determine whether or not.

  When the determination result by the second determining unit 10k is negative, the second flag assigning unit 101 assigns a flag indicating that one cell is not ejected to the second image data. ..

  The first image data acquisition unit 10e, the first determination unit 10f, and the first flag addition unit 10g configure a first selection unit described in the claims, and a second image data acquisition unit. The unit 10j, the second determination unit 10k, and the second flag adding unit 10l constitute a second selection unit described in the claims.

(First Modification of Control Unit 10)
In the first modified example of the control unit 10, when the determination result by the first determination unit 10f is negative (that is, when one cell is not aspirated from the tip 332 into the needle 33), the first determination The section 10f may be configured to determine (for example, 3) how many cells are aspirated from the tip 332 into the needle 33. In this case, the control unit 10 of the first modified example associates the first image data with the number (for example, 3) of cells determined by the first determination unit 10f to associate the first image data with the first association unit (see FIG. 2 is provided instead of the first flag imparting unit 10g.

  The method by which the first associating unit associates the first image data with the number of cells is not limited. For example, (1) the first associating unit creates a database that associates the first image data with the number of cells, and stores the database in a storage medium of a computer. The first associating unit may be configured to update the database each time the first determining unit 10f determines the number of cells. Further, for example, the first associating unit is configured to add the number of cells to the header of the first image data every time the first determining unit 10f determines the number of cells. Good.

  Similarly, in the first modification of the control unit 10, when the determination result by the second determination unit 10k is negative (that is, when one cell is not discharged from the tip 332 into the PCR tube Pij), The second determination unit 10k may be configured to determine (for example, 3) how many cells are ejected from the tip 332 into the PCR tube Pij. In this case, the control unit 10 of the first modified example associates the second image data with the number of cells (for example, three) determined by the second determination unit 10k (for example, 3). 2 is provided in place of the second flag imparting unit 101. The method by which the second associating unit associates the second image data with the number of cells is not limited to the same as in the case of the first associating unit.

(Second Modification of Control Unit 10)
The first determination unit 10f omits the step of determining whether or not one cell is sucked from the tip 332 into the needle 33, and determines how many cells are sucked from the tip 332 into the needle 33. It may be configured to determine whether there is. Similarly, the second determination unit 10k omits the step of determining whether or not one cell is ejected from the tip 332 into the PCR tube Pij, and simply ejects the cell ejected from the tip 332 into the PCR tube Pij. May be configured to determine how many. In these cases, the first associating unit associates with the first image data the number of cells (for example, 3) determined by the first determining unit 10f, and the second associating unit, The number of cells determined by the second determination unit 10k (for example, 3) may be associated with the second image data.

  Note that the first associating unit associates the first image data with the number of cells, and the second associating unit associates the second image data with the number of cells. The method is not limited as in the case of the first modification described above.

(Third Modification of Control Unit 10)
In the above-described first modification, the first determination unit 10f determines the number of cells and gives the first flag only when the determination result by the first determination unit 10f is negative. The section 10g has been described as relating the number of cells to the first image data. However, regardless of the determination result by the first determination unit 10f, the first determination unit 10f determines the number of cells, and the first flag imparting unit 10g determines that the first image data is It may be configured to correlate the number of cells.

  Similarly, regardless of the determination result by the second determination unit 10k, the second determination unit 10k determines the number of cells, and the second flag attachment unit 10l determines the second image data. , May be configured to correlate the number of cells.

  Note that the first associating unit associates the first image data with the number of cells, and the second associating unit associates the second image data with the number of cells. The method is not limited as in the case of the first modification described above.

(Summary of First Embodiment)
The cell delivery system 1 according to the first embodiment sucks one or more cells from a first container (a petri dish SH) in which a plurality of cells are housed, and respectively replenishes each of the one or more cells. When the needle 33 that discharges into each of the one or more second containers (PCR tubes Pij) and the tip 332 of the needle 33 is located in the first container (dishes SH), the vicinity of the tip 332 is observed. When the tip 332 is located in the first microscope (microscope 11) and the second container (PCR tube Pij) or above the opening AP of the second container (PCR tube Pij), And a second microscope (microscope 13) for observing.

  According to the above configuration, the first microscope (microscope 11) for observing cells in the first container (dishes SH) and the second microscope for observing cells in the second container (PCR tube Pij). Since the (microscope 13) can be prepared separately, it is possible to prepare a microscope having a configuration suitable for each application as each of the first microscope (microscope 11) and the second microscope (microscope 13). it can. Therefore, the cell transport system 1 is a cell transport system 1 that transports cells from the first container (the dish SH) to the second container (PCR tube Pij), and the cells are stored in the first container (the dish SH). In addition to when picking cells, a clear image can be obtained when cells are ejected in the second container (PCR tube Pij).

  In addition, the cell transport system 1 according to the first embodiment has at least part of the position of the second microscope (microscope 13) when the tip 332 is located in the second container (PCR tube Pij). It is preferable to further include a first adjusting mechanism (optical stage 43) for adjusting

  In the cell analysis, as the second container, for example, a container such as a PCR tube Pij having a deep depth with respect to the size of the opening AP of the container or a non-uniform bottom thickness is adopted. There are cases. On the bottom surface of the commercially available PCR tube Pij, there is a protruding portion generated when the PCR plate PP is injection-molded.

  When the vicinity of the tip 332 of the needle 33 located in the PCR tube Pij is observed using the second microscope (microscope 13), the contrast in the second container (PCR tube Pij) is too strong. For some reasons, a clear image containing cells may not be obtained. In such a case, the cell transport system 1 can adjust the position of at least a part of the second microscope (microscope 13) by using the first adjustment mechanism (optical stage 43). A clear image including cells can be easily obtained in the container (PCR tube Pij).

  In addition, the cell transport system 1 according to the first embodiment adjusts the position of the second container (PCR tube Pij) when the tip 332 is located inside the second container (PCR tube Pij). A second adjusting mechanism (stage 22) may be further provided.

  In addition, when a clear image including cells cannot be obtained because the contrast in the second container (PCR tube Pij) is too strong, the second adjustment mechanism (optical stage 43) is used instead of the second adjustment mechanism. The position of the second container (PCR tube Pij) may be adjusted by using the adjusting mechanism (stage 22). Even if the second adjusting mechanism (stage 22) is used, a clear image including cells can be easily obtained in the second container (PCR tube Pij).

  In addition, the cell delivery system 1 according to the first embodiment includes a third adjustment mechanism (cell delivery that adjusts the position of the tip 332 when the tip 332 is located in the second container (PCR tube Pij). The robot 30) may be further provided.

  In addition, when a clear image including cells cannot be obtained because the contrast in the second container (PCR tube Pij) is too strong, the first adjustment mechanism (optical stage 43) and the second adjustment are performed. Instead of the mechanism (stage 22), the position of the tip 332 may be adjusted by using the third adjustment mechanism (cell transfer robot 30). Even if the third adjusting mechanism (cell transfer robot 30) is used, a clear image including cells can be easily obtained in the second container (PCR tube Pij).

  When the position of the tip 332 is adjusted by using the cell transfer robot 30, the arm control unit 10a stores the position of the tip 332 before the adjustment in the storage unit, and the second container (for example, PCR tube) When inserting the tip 332 into Pij + 1), it is preferable to control the arm 32 so that the tip 332 is positioned at the position of the tip 332 before adjustment stored in the storage unit.

  In addition, the cell transfer system 1 according to the first embodiment adjusts the position of at least one of the light sources 17a, 17b, and 17c that illuminates the second container (PCR tube Pij) with illumination light. The adjusting mechanism may be further provided.

  In addition, when a clear image including cells cannot be obtained because the contrast in the second container is too strong, the first to third adjustment mechanisms (optical stage 43, stage 22, and cell transfer robot). Instead of 30), the position of the light source may be adjusted by using a fourth adjusting mechanism (for example, a mount including a magnet base and a hinge). Even if the fourth adjusting mechanism (for example, a mount including a magnet base and a hinge) is used, a clear image including cells can be easily obtained in the second container (PCR tube Pij).

  In addition, the cell transport system 1 according to the first embodiment includes a plurality of light sources 17a, 17b, and 17c, which are arranged at different positions and irradiate the second container (PCR tube Pij) with illumination light. A second selection unit 10h that selects a light source to be turned on from the plurality of light sources 17a, 17b, and 17c may be further included.

  According to the above configuration, the cell transport system 1 has the same effects as those provided with the fourth adjusting mechanism (for example, the mount including the magnet base and the hinge).

  In addition, in the cell transport system 1 according to the first embodiment, it is preferable that the second microscope (microscope 13) further includes a switching mechanism (for example, a zoom lens) for switching the magnification.

  According to the above configuration, the operator can easily switch the magnification when observing the inside of the second container (PCR tube Pij). For example, if the operator wants to observe a large area in the second container (PCR tube Pij), the operator can select a relatively low magnification, and the operator can select a narrow magnification in the second container (PCR tube Pij). If the operator wants to observe the area, the operator can select a relatively high magnification. Therefore, the cell transport systems 1 and 2 can improve the usability for the operator.

  Further, in the cell delivery system 1 according to the first embodiment, the one or more second containers are a plurality of second containers, and each of the plurality of second containers is in a matrix form. The plurality of PCR tubes Pij arranged in the plurality of PCR tubes Pij, the needle 33 sucks a plurality of cells from the first container (the petri dish SH), and separates each of the plurality of cells from the plurality of PCR tubes Pij. It is preferable to discharge each.

  According to the above configuration, it is possible to suck a plurality of cells using the needle 33 and then discharge one cell to each of the plurality of PCR tubes Pij. For example, it is possible to suck eight cells from inside the dish SH using the needle 33 and then discharge one cell into each of the PCR tubes P31 to P38 shown in FIG. Therefore, when a large number of cells are isolated and dispensed into the PCR tube Pij one by one, the state in which the tip 332 is located inside the first container (dishes SH) and the inside of the second container (PCR tube Pij). It is possible to reduce the number of times (for example, from 8 times to 1 time in the above case) of switching between the state in which the tip 332 is positioned at the position. Therefore, the labor and time required for isolating and dispensing a large number of cells into the PCR tube Pij can be reduced.

  In the PCR plate PP, the distance between the adjacent PCR tubes Pij and PCR tubes Pij + 1 is a predetermined pitch Px, and the distance between the adjacent PCR tubes Pij and PCR tubes Pi + 1j is the predetermined pitch Py. Is. Therefore, in this case, every time a cell is ejected from the tip 332 located in the PCR tube Pij, the arm control unit 10a controls the arm 32 so as to retract the needle 33 upward, and the first stage control is performed. The unit 10d controls the stage mechanism 22c to move the outer frame 22a to the left by the pitch Px so that the next PCR tube Pij + 1 comes to a predetermined position, and the arm control unit 10a causes the needle 33 to move. The arm 32 may be controlled so that the tip 332 is located in the PCR tube Pij + 1 by moving the arm 32 downward. By configuring the respective functional blocks of the control unit 10 as described above, it is possible to automate (mechanize) the process of continuously discharging cells into each of the plurality of PCR tubes Pij.

  In addition, in the cell transport system 1 according to the first embodiment, a first optical axis (optical axis AL1) that is an optical axis of the first microscope (microscope 11) and the second microscope (microscope 13). The second optical axis (optical axis AL2), which is the optical axis of, is located at a different position from each other, and is on the first optical axis (optical axis AL1) or the second optical axis (optical axis AL2). It is preferable to further include a moving mechanism (cell transfer robot 30) that moves the needle 33 so that the tip 332 is positioned at the position.

  According to the above configuration, since the first optical axis (optical axis AL1) and the second optical axis (optical axis AL2) are located at different positions, the position of the first container (dishes SH) And, without exchanging the position of the second container (PCR tube Pij), the state where the tip 332 is positioned in the first container (dishes SH), and the tip 332 is positioned in the second container (PCR tube Pij). You can switch between the position and the position. Therefore, when switching between the above-described two states, the liquid (culture liquid CF1) contained in the first container (dishes SH) and the liquid (culture liquid CF2) contained in the second container (PCR tube Pij). It is possible to prevent spillage, and to suppress the time and effort of searching for cells again due to movement of cell positions.

  In addition, in the cell transport system 1 according to the first embodiment, the first microscope (microscope 11) is imaged by a plurality of lenses (lens group 11a) and the plurality of lenses (lens group 11a). A first image sensor (image sensor 11b) that generates first image data representing an image; and a first image sensor that acquires the first image data from the first image sensor (image sensor 11b). An image data acquisition unit 10e, a first determination unit 10f that determines whether or not one cell is aspirated from the tip 332 into the needle 33 with reference to the image represented by the first image data, If the result of the determination made by the first determining unit 10f is negative, a first flag assigning unit 10g that assigns a flag indicating that one cell is not aspirated to the first image data is provided. First control unit provided Further comprising a part of the control unit 10), it is preferable.

  According to the above configuration, the control unit 10 of the computer determines, on behalf of the operator, whether or not one cell has been sucked into the needle 33 from the tip 332. Therefore, the labor and time required to isolate a large number of cells one by one from the first container can be reduced. In addition, when one cell is not sucked from the tip 332 into the needle 33, the first flag imparting unit 10g adds a flag indicating that one cell is not sucked to the first image data. Attach. Therefore, when performing cell analysis, it is possible to specify how many cells were aspirated by the needle 33 when the cells were aspirated from each first container (the petri dish SH) to the needle 33.

  In the cell transport system 1 according to the first embodiment, the second microscope (13) has a plurality of lenses (lens group 13a) and an image formed by the plurality of lenses (lens group 13a). And a second image sensor (image sensor 13b) that generates second image data representing the second image data, and obtains the second image data from the second image sensor (image sensor 13b). Second, referring to the data acquisition unit 10j and the image represented by the second image data, it is determined whether or not one cell is ejected from the tip 332 into the second container (PCR tube Pij). If the determination results of the determination unit 10k and the second determination unit 10k are negative, a second flag that indicates that one cell is not ejected is added to the second image data. A flag giving unit 10l, A second control unit having further comprising a (part of the control unit 10), it is preferable.

  According to the above configuration, the control unit 10 of the computer determines on behalf of the operator whether or not one cell has been discharged from the tip 332 into the second container (PCR tube Pij). Therefore, it is possible to reduce the labor and time required for dispensing a large number of cells one by one into the second container (PCR tube Pij). Further, when one cell is not ejected from the tip 332 into the second container (PCR tube Pij), the second flag imparting unit 10l displays a flag indicating that one cell is not ejected. Attached to image data of 2. Therefore, when performing cell analysis, when cells are ejected into each second container (PCR tube Pij), it is determined whether or not one cell is ejected into the second container (PCR tube Pij). Can be specified.

  An algorithm for the first determination unit 10f to determine whether or not one cell has been sucked from the tip 332 into the needle 33, and one cell to be ejected from the tip 332 into the second container (PCR tube Pij). As each of the algorithms for the second determining unit 10k to determine whether or not the existing algorithm (for example, Takashi Kamatani, et. Al., "Construction of a system using a deep learning algorithm to count cell numbers in nanoliter. wells for viable single-cell experiments ", SCIENTIFIC REPORTS, 7: 16831, December 4, 2017,) can be appropriately adopted.

  In addition, in the cell transport system 1 according to the first embodiment, the first microscope (microscope 11) is imaged by a plurality of lenses (lens group 11a) and the plurality of lenses (lens group 11a). A first image sensor (image sensor 11b) that generates first image data representing an image; and a first image sensor that acquires the first image data from the first image sensor (image sensor 11b). An image data acquisition unit 10e, a first determination unit 10f that determines the number of cells aspirated from the tip 332 into the needle 33 by referring to the image represented by the first image data, and the first determination unit 10f. The image processing apparatus may further include a first control unit including a first associating unit (not shown) that associates the number of cells determined by the first determining unit 10f with the image data.

  According to the above configuration, the control unit 10 of the computer determines the number of cells aspirated from the tip 332 into the needle 33 on behalf of the operator. Therefore, the labor and time required to isolate a large number of cells one by one from the first container can be reduced. The first associating unit associates the number of cells determined by the first determining unit 10f with the first image data. Therefore, when performing cell analysis, it is possible to specify the number of cells aspirated by the needle 33 when the cells are aspirated from each of the first containers (the petri dish SH) to the needle 33.

  In the cell transport system 1 according to the first embodiment, the second microscope (13) has a plurality of lenses (lens group 13a) and an image formed by the plurality of lenses (lens group 13a). And a second image sensor (image sensor 13b) that generates second image data representing the second image data, and obtains the second image data from the second image sensor (image sensor 13b). A second determination unit that determines the number of cells discharged from the tip 332 into the second container (PCR tube Pij) with reference to the data acquisition unit 10j and the image represented by the second image data. A second controller (control unit) including 10k and a second associating unit (not shown) that associates the number of cells determined by the second determining unit 10k with the second image data. 10) may be further provided.

  According to the above configuration, the control unit 10 of the computer determines the number of cells discharged from the tip 332 into the second container (PCR tube Pij) on behalf of the operator. Therefore, it is possible to reduce the labor and time required for dispensing a large number of cells one by one into the second container (PCR tube Pij). The second associating unit associates the number of cells determined by the second determining unit 10k with the second image data. Therefore, when performing cell analysis, when the cells are ejected into each second container (PCR tube Pij), the number of cells ejected into the second container (PCR tube Pij) should be specified. You can

  The cell delivery system 1 according to the first embodiment sucks one or more cells from a first container (a petri dish SH) in which a plurality of cells are contained, and removes each of the one or more cells. In the vicinity of the tip 332, the needle 33 discharging into each of the one or more second containers (PCR tubes Pij) and the tip 332 of the needle 33 in the first container (dishes SH) are located. And a second microscope (microscope 13) for observing the vicinity of the tip 332 when the tip 332 is located in the area in which the first microscope (microscope 11) for observing is placed and in the second container (PCR tube Pij). ), And can also be expressed as having.

  In other words, one aspect of the present invention may have a configuration in which the microscope 11 is omitted from the cell transfer system 1 according to the first embodiment.

  According to the configuration, by arranging the first microscope (microscope 11) so that the first optical axis (optical axis AL1) is located in the area, the first container (dishes SH) is separated from the second container. In the case where cells are isolated and dispensed into a container (PCR tube Pij) one by one, in addition to when picking cells in the first container (dishes SH), in the second container (PCR tube Pij) It is possible to obtain a clear image even when the cells are ejected at. As described above, one aspect of the present invention also includes a state in which the first microscope (microscope 11) is not arranged in the cell transport system 1.

  The operator who has obtained the cell transport system 1 in a state in which the first microscope (microscope 11) is not arranged selects the microscope according to the desired specifications as the first microscope (microscope 11) and arranges it in the above area. can do. Therefore, the degree of freedom of the operator can be increased. The first microscope (microscope 11) may be selected from commercially available microscopes or may be self-made.

[Second Embodiment]
The cell delivery system 2 according to the second embodiment of the present invention will be described with reference to FIG. FIG. 4 is a cross-sectional arrow view showing the outline of the cell transport system 2.

  The cell transfer system 2 is a cell transfer system that transfers cells from the Petri dish SH to the PCR tube Pij, like the cell transfer system 1. Therefore, in the present embodiment, the description regarding the petri dish SH and the PCR tube Pij is omitted.

(Configuration of cell transport system 2)
The cell transfer system 2 includes the microscopes 61 and 63, the light sources 67a and 67b, the stage 70, and the cell transfer robot 80 shown in FIG. 4, and an optical stage and a vibration isolation table (not shown in FIG. 4). I have it.

  In the cell transfer system 2, the microscopes 11 and 13, the light sources 17a and 17b, the stages (stages 21 and 22) of the cell transfer system 1, and the cell transfer robot 30 are respectively provided with microscopes 61 and 63 and light sources 67a and 67b. , Stage 70, and cell transfer robot 80.

  The objective lens 62 included in the microscope 61 corresponds to the objective lens 12 of the microscope 61, the housing 64 and the zoom ring 65 included in the microscope 63 correspond to the housing 14 and the zoom ring 15 of the microscope 13, The outer frame 70a included in the stage 70 is a combination of the outer frame 21a of the stage 21 and the outer frame 22a of the stage 22. The plate 72 and the plate 73 included in the stage 70 are the plate 21b of the stage 21. The stage mechanism 74, which corresponds to the plate 22b of the stage 22 and is provided in the stage 70, is a combination of the stage mechanism 21c of the stage 21 and the stage mechanism 22c of the stage 22 and is provided in the cell transfer robot 80. The main shaft 81, the arm 82, and the needle 83 that are installed Of the main shaft 31, corresponding to the arm 32, and the needle 33.

  In the cell transport system 1, the state in which the tip 332 of the needle 33 is located in the dish SH and the state in which the tip 332 is located in the PCR tube Pij are set without moving each of the dish SH and the PCR tube Pij. The transfer robot 30 is switched by moving the position of the needle 33. In other words, in the cell transfer system 1, the optical axis AL1 of the microscope 11 and the optical axis AL2 of the microscope 13 are located at different positions, and the cell transfer robot 30 uses the arm 32 to move the optical axis AL1 on the optical axis AL1. Alternatively, the needle 33 is moved so that the tip 332 is located on the optical axis AL2.

  On the other hand, in the cell delivery system 2, the state in which the tip 332 of the needle 33 is located in the dish SH and the state in which the tip 332 is located in the PCR tube Pij are set without moving the position of the needle 83 in the xy plane. , 70 is used to move the respective positions of the Petri dish SH and the PCR tube Pij to perform switching. In other words, in the cell transport system 2, each of the microscope 61 and the microscope 63 shares a common optical axis AC that is one optical axis, and the stage 70 that is the moving mechanism described in the claims is: At least the Petri dish SH and the PCR tube Pij are moved so that the Petri dish SH or the PCR tube Pij is positioned on the common optical axis AC.

  The cell transport system 2 further includes a mirror 68 that switches the lower end of the common optical axis AC to the microscope 61 side or the microscope 63 side so that the microscope 61 and the microscope 63 share the common optical axis AC. .. The mirror 68 has a rotation axis A68 along the y-axis direction, and switches the lower end of the common optical axis AC to the microscope 61 side or the microscope 63 side by rotating around the axis of the rotation axis A68. ..

  Further, the microscope 63 is supported by the optical stage (for example, the optical stage 43 shown in FIG. 3) which is an example of the first adjusting mechanism described in the claims, like the microscope 13.

  The stage 70 is a stage for mounting the dish SH and the PCR plate PP. The outer frame 71 supports the plate 72 and the plate 73, and is also supported by the stage mechanism 74. The position of the outer frame 71 can be adjusted by using the stage mechanism 74. The stage mechanism 74 is configured similarly to the stage mechanism 21c or the stage mechanism 22c of the cell transfer system 1.

  The cell transfer robot 80 includes a main shaft 81, an arm 82, and a needle 83. The cell transfer robot 80 further includes a pump not shown in FIG.

  Each of the main shaft 81, the arm 82, and the needle 83 is configured similarly to the main shaft 31, the arm 32, and the needle 33 of the cell transfer system 1, respectively. However, in the cell transport system 2, since it is not necessary to move the position of the tip of the needle 83 (corresponding to the tip 332 of the needle 33) in the xy plane, the main shaft 81 does not have to rotate around its central axis. Good. That is, it is sufficient that the cell transfer robot 80 can move at least the position of the tip of the needle 83 up and down in the direction of arrow A8 along the z-axis direction.

(Summary of Second Embodiment)
The cell transport system 2 according to the second embodiment includes a first microscope (microscope 61) and a second microscope (microscope 63).

  Further, it is preferable that the cell transport system 2 according to the second embodiment further includes a first adjusting mechanism (optical stage) that adjusts at least a part of the second microscope (microscope 63).

  The cell transfer system 2 according to the second embodiment may further include a second adjustment mechanism (stage 70) that adjusts the position of the second container (PCR tube Pij).

  The cell delivery system 2 according to the second embodiment may further include a third adjustment mechanism (cell delivery robot 80) that adjusts the position of the tip of the needle 83.

  In addition, the cell transfer system 2 according to the second embodiment is a fourth adjustment that adjusts the position of at least one of the light sources 67a and 67b that illuminates the second container (PCR tube Pij) with illumination light. A mechanism may be further provided.

  In addition, the cell transport system 2 according to the second embodiment includes a plurality of light sources 67a and 67b arranged at different positions for illuminating the second container (PCR tube Pij) with illumination light. A second selection unit (corresponding to, for example, the second selection unit 10h shown in FIG. 2) for selecting a light source to be turned on from the light sources 67a and 67b.

  In addition, in the cell transport system 2 according to the second embodiment, it is preferable that the second microscope (microscope 63) further includes a switching mechanism (for example, a zoom lens) for switching the magnification.

  In addition, in the cell delivery system 2 according to the second embodiment, the needle 83 sucks a plurality of cells from the first container (the petri dish SH), and the plurality of cells are respectively attached to the plurality of cells. It is preferable to discharge each of the PCR tubes Pij.

  In addition, in the cell transport system 2 according to the second embodiment, the first microscope (microscope 61) has a plurality of lenses (corresponding to the lens group 11a shown in FIG. 2) and an image formed by the plurality of lenses. A first image sensor (corresponding to the image sensor 11b shown in FIG. 2) that generates first image data representing the captured image, and obtains the first image data from the first image sensor. With reference to the first image data acquisition unit (corresponding to the first image data acquisition unit 10e shown in FIG. 2) and the image represented by the first image data, the inside of the needle 83 from the tip of the needle 83 In the case where the determination result by the first determination unit (corresponding to the first determination unit 10f shown in FIG. 2) that determines whether or not one cell is aspirated is For the first image data, one cell is aspirated A first control unit (a part of the control unit 10 shown in FIG. 2) including a first flag addition unit (first flag addition unit 10g shown in FIG. 2) that gives a flag indicating that there is no (Corresponding to the above) is further provided.

  Further, in the cell delivery system 2 according to the second embodiment, the second microscope (63) forms an image with a plurality of lenses (corresponding to the lens group 13a shown in FIG. 2) and the plurality of lenses. A second image pickup device (corresponding to the image pickup device 13b shown in FIG. 2) that generates second image data representing an image, and obtains the second image data from the second image pickup device. By referring to the second image data acquisition unit (corresponding to the second image data acquisition unit 10j shown in FIG. 2) and the image represented by the second image data, the second image from the tip of the needle 83 is referred to. A second determination unit (corresponding to the second determination unit 10k shown in FIG. 2) that determines whether or not one cell has been discharged into the container (PCR tube Pij), and the determination result by the second determination unit Is no, for the second image data, A second control unit (control unit 10) including a second flag giving unit (corresponding to the second flag giving unit 101 shown in FIG. 2) that gives a flag indicating that two cells are not ejected. (Part of) is further provided.

  In addition, in the cell transport system 2 according to the second embodiment, the first microscope (microscope 61) has a plurality of lenses (corresponding to the lens group 11a shown in FIG. 2) and an image formed by the plurality of lenses. A first image sensor (corresponding to the image sensor 11b shown in FIG. 2) that generates first image data representing the captured image, and obtains the first image data from the first image sensor. With reference to the first image data acquisition unit (corresponding to the first image data acquisition unit 10e shown in FIG. 2) and the image represented by the first image data, the inside of the needle 93 from the tip of the needle 83 A first determination unit (corresponding to the first determination unit 10f shown in FIG. 2) that determines the number of cells sucked into the first determination unit, and the first determination unit determines the first image data. A first association unit (not shown) for associating the number of cells A first control unit (corresponding to a portion of the controller 10 shown in FIG. 2) may further comprise a with.

  Further, in the cell transport system 2 according to the second embodiment, the second microscope (63) includes a plurality of lenses (corresponding to the lens group 13a shown in FIG. 2) and the plurality of lenses (lens group 13a). ), And a second image sensor (corresponding to the image sensor 13b shown in FIG. 2) that generates second image data representing the image formed by The tip of the needle 83 is referred to with reference to the second image data acquisition unit (corresponding to the second image data acquisition unit 10j shown in FIG. 2) acquired from the image sensor and the image represented by the second image data. A second determination unit (corresponding to the second determination unit 10k shown in FIG. 2) for determining the number of cells discharged from the above into the second container (PCR tube Pij), and the second image data. In contrast, the number of the cells determined by the second determination unit is Second associating unit attaching communicating with (not shown), the second control unit may further comprise a (controller 10) having a.

  The cell transfer system 2 according to the second embodiment also includes a region in which the first microscope (microscope 61) for observing the vicinity of the tip of the needle 83 is arranged and a second microscope (microscope 63). Can be expressed.

  Since the cell delivery system 2 according to the second embodiment is configured as described above, the cell delivery system 2 has the same effect as the cell delivery system 1.

  In addition, in the cell transport system 2 according to the second embodiment, each of the first microscope (microscope 61) and the second microscope (microscope 63) shares a common optical axis AC that is one optical axis. At least the first container (the dish SH) and the second container (the dish SH) so that the first container (the dish SH) or the second container (the PCR tube Pij) is located on the common optical axis AC. A moving mechanism (stage 70) for moving the container (PCR tube Pij) may be further provided.

  According to the above configuration, since the first microscope (microscope 61) and the second microscope (microscope 63) share the common optical axis AC that is one optical axis, the first optical axis (optical axis The space for arranging the two microscopes can be narrowed as compared with the cell transport system 1 in which AL1) and the second optical axis (optical axis AL2) are located at different positions. That is, the cell delivery system 2 can be made more compact than the cell delivery system 1.

[Third Embodiment]
(Example of realization by software)
Control block of the control unit 10 of the cell transport system 1 (particularly, the first image data acquisition unit 10e, the first determination unit 10f, the first flag attachment unit, the second image data acquisition unit 10j, and the second determination). The unit 10k and the second flag giving unit 10l may be realized by a logic circuit (hardware) formed on an integrated circuit (IC chip) or the like, or may be realized by software.

  In the latter case, the control unit 10 of the cell transfer system 1 includes a computer that executes the instructions of a program that is software that realizes each function. The computer includes, for example, one or more processors and a computer-readable recording medium that stores the program. Then, in the computer, the processor reads the program from the recording medium and executes the program to achieve the object of the present invention. As the processor, for example, a CPU (Central Processing Unit) can be used. As the recording medium, a "non-transitory tangible medium" such as a ROM (Read Only Memory), a tape, a disk, a card, a semiconductor memory, a programmable logic circuit, or the like can be used. Further, a RAM (Random Access Memory) for expanding the program may be further provided. The program may be supplied to the computer via any transmission medium (communication network, broadcast wave, etc.) capable of transmitting the program. Note that one aspect of the present invention can also be realized in the form of a data signal embedded in a carrier wave, in which the program is embodied by electronic transmission.

  The control block of the control unit of the cell transfer system 2 is the same as the control block of the control unit 10 of the cell transfer system 1.

  The present invention is not limited to the above-described embodiments, and various modifications can be made within the scope of the claims, and the embodiments obtained by appropriately combining the technical means disclosed in the different embodiments. Is also included in the technical scope of the present invention.

1, 2 cell transport system SH Petri dish (first container)
PP PCR plate Pij, P31 to P38 PCR tube (second container)
AP opening 10 control part 10e, 10j 1st image data acquisition part, 2nd image data acquisition part 10f, 10k 1st determination part, 2nd determination part 10g, 10l 1st flag provision part, 2nd Flagging unit 10h Second selection unit (selection unit)
11,61 Microscope (first microscope)
11a lens group (plural lenses)
11b Image sensor AL1 optical axis (first optical axis)
13,63 Microscope (second microscope)
13a lens group (plurality of lenses)
13b Image sensor AL2 optical axis (second optical axis)
AC common optical axes 16a, 16b, 17a, 17b, 17c, 67a, 67b Light source 21 Stage 22 Stage (second adjusting mechanism)
70 stages (second adjusting mechanism, moving mechanism)
30 Cell transfer robot (3rd adjusting mechanism, moving mechanism)
80 Cell transfer robot (3rd adjustment mechanism)
31, 81 Spindle 32, 82 Arm 33, 83 Needle 43 Optical stage (first adjusting mechanism)

Claims (15)

A needle that aspirates one or more cells from the first container containing a plurality of cells and discharges each of the one or more cells into each of the one or more second containers;
A first microscope for observing the vicinity of the tip when the tip of the needle is located in the first container;
A cell transfer system, comprising: a second microscope that observes the vicinity of the tip when the tip is located in the second container or above the opening of the second container.   The cell delivery system according to claim 1, further comprising a first adjusting mechanism that adjusts the position of at least a part of the second microscope when the tip is located in the second container.   The cell delivery system according to claim 1 or 2, further comprising a second adjusting mechanism that adjusts the position of the second container when the tip is located in the second container.   The cell delivery system according to claim 1, further comprising a third adjustment mechanism that adjusts the position of the tip when the tip is located in the second container.   The cell delivery system according to any one of claims 1 to 4, further comprising a fourth adjustment mechanism that adjusts a position of a light source that illuminates the second container with illumination light. A plurality of light sources arranged in mutually different places for irradiating the second container with illumination light;
The cell transport system according to claim 1, further comprising a selection unit that selects a light source to be turned on from the plurality of light sources.   The cell delivery system according to claim 1, wherein the second microscope further includes a switching mechanism that switches a magnification. The one or more second containers are a plurality of second containers,
Each of the plurality of second containers is a plurality of PCR tubes arranged in a matrix,
8. The needle aspirates a plurality of cells from the first container, and discharges each of the plurality of cells into each of the plurality of PCR tubes. 8. Cell transport system. The first optical axis, which is the optical axis of the first microscope, and the second optical axis, which is the optical axis of the second microscope, are located at different positions.
9. The moving mechanism according to claim 1, further comprising a moving mechanism that moves the needle so that the tip is located on the first optical axis or the second optical axis. Cell transport system. Each of the first microscope and the second microscope shares a common optical axis which is one optical axis,
The moving mechanism for moving at least the first container and the second container so as to position the first container or the second container on the common optical axis is further provided. 8. The cell delivery system according to any one of 8 above. The first microscope includes a plurality of lenses, and a first image sensor that generates first image data representing an image formed by the plurality of lenses,
A first image data acquisition unit that acquires the first image data from the first image sensor;
A first determination unit that determines whether or not one cell is aspirated from the tip into the needle with reference to the image represented by the first image data;
A first flag assigning unit that assigns a flag indicating that one cell is not aspirated to the first image data when the determination result by the first determining unit is negative. The cell delivery system according to any one of claims 1 to 10, further comprising a provided first control unit. The second microscope includes a plurality of lenses, and a second image sensor that generates second image data representing an image formed by the plurality of lenses,
A second image data acquisition unit for acquiring the second image data from the second image sensor;
A second determination unit that determines whether or not one cell is discharged from the tip into the second container with reference to the image represented by the second image data;
A second flag assigning unit that assigns a flag indicating that one cell is not ejected to the second image data when the determination result by the second determining unit is negative. The cell delivery system according to any one of claims 1 to 10, further comprising a second control unit provided. The first microscope includes a plurality of lenses, and a first image sensor that generates first image data representing an image formed by the plurality of lenses,
A first image data acquisition unit that acquires the first image data from the first image sensor;
A first determination unit that determines the number of cells aspirated from the tip into the needle with reference to the image represented by the first image data;
The first controller further comprising: a first associating unit that associates the number of cells determined by the first determining unit with the first image data. 10. The cell delivery system according to any one of 10. The second microscope includes a plurality of lenses, and a second image sensor that generates second image data representing an image formed by the plurality of lenses,
A second image data acquisition unit for acquiring the second image data from the second image sensor;
A second determination unit that determines the number of cells discharged from the tip into the second container with reference to the image represented by the second image data;
The second control unit further comprising: a second associating unit that associates the number of cells determined by the second determining unit with the second image data. 10. The cell delivery system according to any one of 10. A needle for sucking one or more cells from a first container containing a plurality of cells and discharging each of the one or more cells into each of the one or more second containers When,
A region for arranging a first microscope for observing the vicinity of the tip when the tip of the needle is located in the first container,
A second microscope for observing the vicinity of the tip when the tip is located in the second container.

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