JPH0731456A - Cell fusion apparatus - Google Patents

Abstract

PURPOSE:To obtain a cell fusion apparatus provided with a transferring means, an association means, a fusing means, a post-treatment means and an observation means, capable of automatically performing the fusing operation and getting a large amount of fused cells and useful in the field of biotechnology, etc. CONSTITUTION:This cell fusion apparatus is provided with a controller 19 to control the operation of a transferring means to transfer a cell with light radiating from a laser emitter 18, an association means to apply a DC pulse with an electrode 1, a fusing means and a post-treatment means. An observation means 16 transmits the image in the visual field to the controller 19 as an image data. The controller 19 has a function to analyze the image data and a function to control the operation of one or more means among the transfer means, the association means, the fusion means and the post-treatment means according to the analyzed result.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a cell fusion device for fusing cells in the field of biotechnology.

[0002]

2. Description of the Related Art Conventionally, as this type of cell fusion device,
For example, there is one as shown in FIG. The cell fusion device shown in FIG. 2 includes a microscope 21 for observing cells, a pair of electrodes 22 for applying a voltage to the cells, a power supply 23 for applying a voltage to the electrodes 22, a stand 24, and And an illumination 25 for illuminating the field of view of the microscope 21. The stand 24 supports the microscope 21 and the illumination 25. The stand 24 also includes a stage 29 and a horizontal member 26. The electrode 22 is rod-shaped, has a thin tip, and is movably attached to a horizontal member 26 connected to a stand 24.

An operator first places a petri dish 27 containing a suspension in which cells 28 to be fused are suspended on a stage 29 of a stand 24. Next, while observing the cells with the microscope 21, the cells are pushed and moved by the electrode 22, and the cells 28 to be fused are brought into contact with each other. If a DC pulse is applied to the electrode 22 while the cells are associated with each other at the electrode 22, the cells will fuse (Senda et.al., Plant and Cell).
Physiol. 20 (7) 1441-1443 (1979)).

[0004]

However, in such a conventional cell fusion device, a skilled operator must perform the fusion by a work, and even one fusion operation takes time. Also, a great deal of labor is required. Therefore, it is not realistic to obtain a large amount of fused cells with such a cell fusion device. The fusion operation here is
It refers to steps including cell selection, delivery, association, fusion, and post-treatment.

Therefore, an object of the present invention is to provide a cell fusion device capable of automatically executing the above fusion operation.

[0006]

In order to achieve the above object, the present invention provides at least one suspension chamber for holding cells for fusion, and a fusion chamber for fusing cells. A collection chamber for collecting the cells after the fusion treatment, an observation means for observing the inside of the visual field, a transportation means for transporting the cells, an association means for associating the cells, a fusion means for fusing the cells, and a fusion treatment after the fusion processing. A cell fusion apparatus having a post-treatment means for performing a post-treatment, comprising a control device for controlling execution of the transport means, the association means, the fusion means, and the post-treatment means, and the observation means. Has a function of transmitting an image within the field of view as image data to the control device, the control device having a function of analyzing the image data, and the conveying means according to the analysis result,
There is provided a cell fusion device having a function of controlling the operation of at least one of the association means, the fusion means, and the post-processing means.

Further, in the present invention, the function of analyzing the image data is to detect the contour and / or position of cells,
There is provided a cell fusion device having a function of making a judgment based on the shape and / or position of the contour.

Further, the transporting means has a transporting mechanism using a light trapping phenomenon, in which the cells are moved by irradiating the cells with light and moving the irradiation position of the spot of the light, and the image data is transferred. The analyzing function has a function of detecting the irradiation position and / or the contour of the light spot, and the control device has a function of controlling the conveying means based on the irradiation position and / or the contour. A characteristic cell fusion device is provided.

[0009]

The cell fusion apparatus of the present invention pattern-recognizes image data from a monitor camera attached to a microscope to obtain contours and positions of cells, organ walls, and spots of laser light. Furthermore, by analyzing the contours obtained, for selection of cells to be fused, judgment of success or failure of association and fusion, control of transport speed and transport position by optical trapping, adjustment of spot size of laser light, for association The strength of the high-frequency voltage applied to the device is adjusted, and the number of DC pulses applied for fusion is adjusted. As a result, the cell fusion device of the present invention automatically carries out, associates, and fuses cells. Further, the cell fusion device of the present invention can repeatedly execute each of the above processes.

The contour of the cell is obtained as follows. First, a threshold value is set from a histogram for a rectangular area having a size in which cells for which contours are to be included can be set, and cells (areas) are obtained by binarization processing. Alternatively, the contour of the cell is obtained by performing the differential processing or the differential processing of the image in the rectangular area.

Since cells are generally in the shape of a circle, the cell fusion apparatus of the present invention determines the cell having the contour closest to a circle among the cells in the microscopic field as the cell to be fused. Further, since the associated cells and the cells immediately after the fusion have a shape in which a plurality of circles are in contact with each other, the success or failure of the association and the fusion can be recognized by the contours of the cells.

The cell fusion device of the present invention utilizes a light trapping phenomenon and a dielectrophoresis phenomenon for transporting cells. First, the cells are transported from the suspension chamber holding the cell suspension in which they are suspended to the vicinity of the electrodes by the light trapping phenomenon. That is, the cells to be fused are irradiated with laser light to capture the cells, and the irradiation position is moved to convey the captured cells.

The optical trapping phenomenon is a phenomenon in which particles are trapped by light. This is because the momentum of photons changes when light refracts at the interface between the surrounding solvent and cells, so that the momentum of light and object is preserved in both reflection and refraction. This is due to the generation of force in different directions. Since this force has a component in the direction in which the intensity of the laser beam is strong, cells can be trapped near the optical axis of the laser beam. By utilizing this phenomenon, the cells captured by the laser beam can be transported by moving the laser light source and arranged at a desired position.

Next, the cell fusion device of the present invention associates the cells arranged in the vicinity of the electrodes by the dielectrophoresis phenomenon. That is, a high frequency voltage is applied to the electrodes, which causes the cells to approach each other and associate with each other.

The dielectrophoresis phenomenon is a phenomenon in which neutral particles are connected to each other in the electric field direction due to polarization generated in electrically neutral particles in a high-frequency alternating electric field, or neutral particles are attracted to a stronger electric field strength. Since cells are electrically neutral particles, they can be moved and associated by dielectrophoresis.

[0016]

EXAMPLE An example of the present invention will be described below with reference to FIG. The cell fusion device of the present embodiment is provided with an observation unit 16 for observing the state of cells, a cell holding unit 17 including each chamber for holding cells and a flow path between the chambers, and for transporting cells. Laser light generator 1 for generating laser light
8 and a control device 19 that controls each of the above units.

The observation unit 16 has a microscope 12 for observing the state of cells, and a monitor camera 13 such as a CCD camera for transmitting the image obtained by the microscope to the control device 19 as image data. The microscope 12
May be a magnifying glass depending on the size of cells to be fused.

The cell holding unit 17 is a chamber table 7 which is kept horizontal so that the liquid to be held does not flow, and a chamber table which is a moving mechanism for moving the chamber table 7 while keeping it horizontal. And an XYZ stage 8 for use.
The chamber stage XYZ stage 8 has a drive device (not shown).

On the chamber table 7, a pair of electrodes 1 for associating and fusing cells, a suspension chamber 3 for holding a cell suspension, and a fusing chamber 2 for fusing cells.
And a collection chamber 4 for collecting the fused cells,
A waste chamber 5 for collecting cells that have failed to fuse. The width between each chamber is 50 μm and the length is 5 μm.
mm channels are provided. In this example, protoplasts having a diameter of about 30 μm were used. The length of each channel is such that when a high frequency voltage is applied to the electrode 1, cells held in the suspension chamber 3, the collection chamber 4, and the waste chamber 5 cause dielectrophoresis due to the high frequency voltage. , Must be long enough not to be pulled into the fusion chamber 2.

In this embodiment, two suspension chambers 3 are provided to fuse two types of cells. However, the suspension chamber 3 is prepared according to the type of cells to be fused, and when only the same cells are fused, one suspension chamber 3 is sufficient. The fusion chamber 2
Are provided in the gap between the electrodes 1 facing each other. In addition, the cell holding unit 17 is a driving device 6 for the electrode 1.
Have. The driving device 6 has a power source and can generate a high frequency voltage and a DC pulse between the electrodes 1. Further, the drive device 6 can adjust the frequency, amplitude, and duration of the high-frequency voltage and the voltage and duration of the DC pulse.

The gap between the opposing electrodes 1 was 1 mm in this embodiment. Further, in this example, the high frequency voltage was 500 KHz to 1 MHz, the voltage was 1 to 20 V / mm, and the DC pulse was 75 V / mm and 10 μsec.

It is desirable that the chamber base, the chamber, and the walls of the flow path on the side that irradiates the laser light are made of a material that does not absorb or reflect the laser light. In this embodiment, laser light is emitted from under the chamber table. Therefore, the bottom of the chamber base 7 of this embodiment is made of glass.

The laser light generator 18 has a laser light source 9 for generating a laser light, a laser light source XYZ stage 10 for moving the laser light source 9, and a laser light source driving device 11. The laser light source 9
Is a mechanism for generating a laser beam and condensing the laser beam. In this example, a laser beam in the near infrared region was generated by using a semiconductor laser of 30 mW. Further, the laser light source 9 has a collimating mechanism, and further has a condenser lens whose optical axis is adjusted and whose focal length can be changed. The driving device 11 has a power source and controls the generation of laser light by the laser light source 9. Further, the drive device includes an electronic relay, and is controlled by opening and closing the relay according to a signal from the control device 19.

In this embodiment, the laser light generator 18 is
The irradiating laser light is configured to be incident on the bottom surface of the chamber base 7 at an arbitrary angle, and the XYZ stage 10 for a laser light source has a driving device (not shown) and controls the incident angle of the laser light. The laser light source 9 can be moved so that the laser light can be irradiated to an arbitrary position in the chamber while keeping it constant.

In this embodiment, the chamber table XYZ is used.
Although both the stage 8 and the XYZ stage 10 for the laser light source are provided, in the present embodiment, the XY for the laser light source is used.
The Z stage 10 is used to irradiate the laser light to an arbitrary position on the chamber base 7, and the XYZ stage 8 for the chamber base is used to set the visual field of the microscope 12 to an arbitrary position on the chamber base 7. However, the present invention is not limited to the above moving mechanism. Any mechanism can be used as long as it can irradiate the laser beam on any position of the chamber base 7 and can set the visual field of the microscope 12 to any position of the chamber base 7.

The above two XYZ stages 8 and 10 are 3
Each has a micrometer (not shown) for each axis (X-axis, Y-axis, Z-axis) of the direction. By rotating the micrometer, the XYZ stages 8 and 10 are moved to X,
It can be moved linearly in each of the Y and Z axis directions.
The mounting order of the stages in each direction may not be as illustrated.

Further, the above two XYZ stages 8 and 10 are also provided.
Has a drive device (not shown) for rotating the micrometer for each micrometer. The drive is
The micrometer is rotated at an arbitrary rotation angle according to an electric signal (voltage value) from the outside. In this embodiment,
Although the stepping motor is used as the driving device, an ultrasonic motor or the like may be used. The control device 19 can control the movement by the micrometer by sending a signal to the driving device, and the minute movement of the stage and the X, Y, Z
It is possible to perform straight movement in a direction other than each axis, movement along a curve, and the like. In this embodiment, the vertical direction is the Z axis, and of the horizontal directions, the direction of the electric force lines between the electrodes 1 is the X axis, and the direction orthogonal to the electric force lines is the Y axis.

In this embodiment, the drive unit is provided with A phase, B
A two-phase stepping motor having two phases is used and driven by a microstepping method. For example, the A phase and the B phase are driven by microsteps in which a sine wave shifted by a quarter wavelength is digitized. By doing so, the stage can be moved with smoothness, accuracy, and positioning that are proportional to the resolution of the microstep.
The moving speed can be adjusted by the cycle or time width of the microstep. Further, since the position of the chamber table can be accurately grasped, it is possible to accurately move the irradiation position of the laser beam and the visual field position of the microscope 12 in the narrow flow path between the chambers in the narrow flow path between the chambers. it can. For example, the longitudinal direction of the channel is represented by a straight line on the XY plane (a plane horizontal to the bottom surface of the chamber base), and it is possible to easily move along the straight line (parallel) accurately. Furthermore, since the moving speed can be controlled, it is possible to easily change the moving speed arbitrarily according to the ease of moving cells, such as always moving at a constant speed or moving slowly.

The control device 19 includes an interface unit 30 for controlling transmission / reception of signals to / from the above-mentioned units, and an input / output device 34 for receiving an operator's input and outputting a processing result and the like.
And a central processing unit 31 for performing arithmetic processing and a storage unit 32 for holding data. Further, the control device 19
Has an input device 34 that receives an operator's input, and an output device 33 that outputs an image obtained through the monitor camera 13, a processing result, and the like. In this embodiment, since the output device 33 can output image data at high speed, a television monitor is used, but other output means may be used.

The control device 19 controls the application of voltage to the electrode 1, the movement of the visual field position of the microscope by moving the XYZ stage 8 for the chamber base, and the irradiation position of the laser light by driving the XYZ stage 10 for the laser light source. Of the laser light, the control of the irradiation of the laser light from the laser light source 9, and the control of the movement of the focal length of the condenser lens in the laser light source 9.

Control device 1 in the cell fusion device of this embodiment
The outline of the hardware configuration of the part related to the function of 9
As shown in FIG. The controller 19 has a data bus 35.
In this embodiment, a 16-bit logic circuit is used as the data bus 35. Further, the central processing unit 31 of the control unit 19
Has an image processing unit 31a and a calculation processing unit 31b.
The image processing unit 31a and the arithmetic processing unit 31b are classified according to their functions, and in practice, in the present embodiment, both functions are realized on the same hardware. However, hardware dedicated for each function may be prepared. The storage device 32 of the control device 19 has a frame memory (image memory) 32a and can hold image data. The image data held in the frame memory 32a is converted into an analog signal through the analog / digital converter 36 and output to the output device 33. The output of the image data is performed for confirmation by the operator and is not essential for the cell fusion process by the control device 19. Therefore, the output of such image data may be omitted or simplified.

Image data sampled by the monitor camera 13 and converted into a digital signal by the analog / digital converter 36 are input to the controller 19, and
There is operator instruction data received by the input device.
In addition, the output from the control device 19 includes the XY for the chamber base.
There are control data for driving the driving device 8a of the Z stage 8 and the driving device 10a of the XYZ stage 10 for the laser light source, and control data for the driving device 6 of the electrode 1 and the driving device 11 of the laser light source 9. In addition,
A command analyzer 37 is provided between each drive device and the control device 19. This is because each control data is output from the control device 19 in the form of a command, and therefore it is necessary to analyze the command by the command analyzer 37 and actually convert it into drive information of each drive device.

The control device 19 holds the image data collected by the monitor camera 13 in the frame memory 32. Further, the image data held in the frame memory 32 is output as an image to the output device 33. Further, the image processing unit 31a performs image recognition processing on the image data to determine the contour of the cell. The arithmetic processing unit 31b determines which driving device is to be controlled and how to control it based on the determined information such as the contour of the cell, and outputs it to each driving device via the data bus 35.

The cell fusion in this example is carried out as follows. The culture solution is held in advance in the fusion chamber 2, the collection chamber 4, and the disposal chamber 5. The suspension chamber 3 holds a suspension of cells to be fused. In this embodiment, the input device 34
An operator's start instruction inputted via the controller 19 is input.
When is accepted, the following fusion operation is started. Also,
The fusion operation is repeated until an operator's end instruction is input (step 401) or a preset number of fused cells are obtained (step 402).

(1) The control device 19 is an XY for chamber base.
The Z stage 8 is driven (step 403), the chamber base 7 is moved horizontally, and the visual field of the microscope 12 is set inside the suspension chamber 3. Here, the control device 19 determines one cell in the visual field as a target cell to be subjected to fusion (step 404). It should be noted that this decision may be made by the operator observing the image of the output device 33 and making a decision, and may be specified to the control device 19 using the cursor superimposed on the frame memory via the input device 34. The control device 19 may determine itself based on a criterion such as the one in which the contour of the cell is closest to the circle in the visual field. In this embodiment, the latter is used. The contour of the cell is obtained as follows. First, a threshold value is set from a histogram for a rectangular area having a size in which cells for which contours are to be included can be set, and cells (areas) are obtained by binarization processing. Alternatively, the contour of the cell is obtained by performing the differential processing or the differential processing of the image in the rectangular area. The area within this contour is the cell area. The center of gravity of the cell area is the cell position.

(2) Next, the control device 19 provides a monitoring area (window) around the target cell on the image (step 405). That is, a rectangular area (window) is set in the frame memory so that the target cell selected in the process (1) completely enters. This cell area (depending on the cell type, etc., in this embodiment, the diameter is about 30 μm)
A slightly larger rectangular area (in this embodiment, one side is approximately 40
A square (μm square) is provided as a monitoring area. This area is effective for identifying cells and shortening the processing time for contour detection.

(3) The control device 19 turns on the laser light source 9. Furthermore, an XYZ stage 10 for a laser light source
Is driven to move the laser light source 9, align the optical axis of the laser with the target cell determined in (1) in the monitoring area, and further control the focal length of the condensing lens in the laser light source 9. The size of the light spot is adjusted according to the size of the target cell (step 406). The target cells are attracted by the laser light due to the light trapping phenomenon.

(4) The controller 19 detects the flow path in the field of view and the inner wall of the chamber by image processing, and further detects the cell position and the laser light irradiation position (step 407). The laser irradiation position is recognized by determining the area with the highest brightness in the visual field.
Further, since the laser light spot has high brightness, the brightness on the image is reduced by using the red cut filter. Next, the control device 19 causes the cells to effectively perform the dielectrophoresis by the target position (voltage applied to the electrode 1). It is checked whether or not the range has been reached (step 408). If not, the controller 19 drives the chamber XYZ stage 10 to relatively displace the chamber pedestal 7 and the laser light source, and conveys the captured target cells for a certain period of time by optical trapping (step 409). ), The process returns to step 407. As in the moving method described above, if the chamber base 7 is moved and the irradiation position of the laser light is fixed, the irradiation position can be kept within the visual field of the microscope 12 at all times.

In the above-mentioned cell transportation, the control device 19
If the distance between the cell position and the laser light irradiation position is smaller than a preset value, the moving speed of the laser light source (moving speed of the chamber table) is increased (step 409).
a) If it is larger, the moving speed is reduced (step 40).
9b). By doing so, it is possible to move the cells captured by the laser light by adjusting the speed so that the cells fit within the monitoring area.

The control of the moving direction / distance is performed so that the flow path and the inner wall of the chamber are detected by image processing, and the irradiation positions of the cells and the laser light are moved along the detection. As the image processing method, the contour of the inner wall is obtained by the difference or differentiation processing as described in (2), and the inner wall contour is controlled so as to keep the distance between the cells constant, and the parallel to the inner wall. The method of moving to is used.

Since the relative position of the field of view of the microscope 12 with respect to the chamber can be obtained from the rotation of the stepping motor of each stage driving device 8, a course is set in advance, and the relative position is taken into consideration. It is also possible to adopt a method of moving the irradiation position of the cells and the laser light along the set course. Also, a method combining the above two methods can be adopted. That is, the outline is a method in which the outline is moved according to a preset course, and the movement is accurately controlled based on the image processing information for obtaining the contour when the movement is small (when it is moved out of the chamber to the flow path).

(5) If the target position is reached in step 408, the cells have already been transferred to a position where they can be associated with each other by the dielectrophoresis phenomenon, and therefore, it is necessary to further transfer the cells by the optical trapping phenomenon. There is no. Therefore, the control device inspects whether or not there are other cells to be transported by the optical trapping phenomenon, and executes the above (1) to (4) for each of all the fusion chambers 3 (step 4).
10). As a result, all cells to be fused are arranged near the electrode 1.

(6) The control device 19 applies a high frequency voltage between the electrodes 1 (step 411) to cause a dielectrophoresis phenomenon. Further, the control device 19 confirms that the cells to be fused are associated by analyzing the image data (step 412). Usually, cells have a substantially circular shape, so if the outline of a cell has a gourd-like shape with two circles in contact (illustrated in Fig. 5 (a)), the two cells Can be recognized as having a meeting.

If the cells are not yet associated, if the voltage is the upper limit value (step 413), the cells are transported through the discard chamber (step 416), and the process is returned to step 401,
Once again, new cells are obtained from the fusion chamber. If the voltage is not the upper limit value, the control device 19 determines whether or not the cell has been destroyed, based on whether or not the contour of the cell is still circular (step 414). If destroyed,
Similar to the case where the voltage is the upper limit value, the process proceeds to step 416. When the voltage is not destroyed, the voltage is increased and the high frequency voltage is applied again (step 411).

The voltage for dielectrophoresis varies depending on the electric conductivity of the cell suspension. The lower the electric conductivity, the lower the voltage may be. However, it is desirable to apply the high frequency voltage in the range of 0 to 20 V / mm. . When the voltage is higher than 20 V / mm, a water flow is generated in the suspension and the cells sway, so that stable association cannot be obtained. In addition, at a conductivity of 50 μS / cm, a cell association can be induced at a voltage of about 10 V / mm. In this embodiment, starting from 1 V / mm, the voltage is increased by 1 V / mm to obtain 20 V / m.
If no association is obtained even after applying m, it is considered that the cells are attached to the organ wall, so that the cells were discarded and new cells were prepared.

(7) When the cell association is successful, the controller 19 directs the DC pulse between the electrodes 1 (the strength required for fusion is
In this embodiment, a voltage of 75 V / mm and a DC pulse of 10 μsec) is applied, though it varies depending on the type of cells and the state of the suspension (step 417). Next, the controller 19 inspects whether or not the cells are destroyed in the same manner as in step 414 (step 418). If destroyed, the cell is discarded (step 416) and the process returns to step 401.
If not destroyed, check whether the cells have fused (419). Immediately after fusion, the cells are more in contact with each other than the gourd-like shape (Fig. 5 (a)) in which two circles are in contact with each other, which is the outline of two cells associated with each other. Since it has an expanded shape (FIG. 5B), the success or failure of fusion can be recognized by the contour of the cell. If they have not been merged, return to step 417,
The DC pulse is applied again. When they are fused, the fused cells are collected in the collection chamber (step 420) and the process returns to step 401, and the control device 19 controls the above (1) to (1).
The process of (7) is repeated the required number of times. The transportation of the cells to the discard chamber (step 416) and the transportation to the collection chamber (step 420) are performed using the optical trapping phenomenon by the same procedure as the above (1) to (4).

In the above process (7), if the cells do not fuse even if the DC pulse is applied, the cells may be fused or not.
The method of repeatedly applying the DC pulse until the destruction is adopted, but the method of increasing the application time of the DC pulse can be used together. The longer the application time, the easier the cells are to fuse, but if the application time is too long, the probability of cell destruction increases, so the application time is preferably 10 to 50 μsec, and 10 to 30 μsec. Is more preferable.

[0048]

As described above, according to the present invention, there is provided a cell fusion device capable of automatically executing a fusion operation. In the cell fusion device of the present invention, a large number of fused cells can be obtained easily by repeatedly performing the fusion operation.

[Brief description of drawings]

FIG. 1 is an explanatory diagram of a cell fusion device of the present invention.

FIG. 2 is an explanatory view of a conventional cell fusion device.

FIG. 3 is a hardware configuration diagram of the cell fusion device of the present invention.

FIG. 4 is a flowchart showing a cell fusion process of the control device.

FIG. 5 is an explanatory diagram showing contours of associated cells and fused cells.

[Explanation of symbols]

1: electrode, 2: fusion chamber, 3: suspension chamber, 4: collection chamber, 5: waste chamber, 6:
Electrode drive device, 7: chamber stage, 8: chamber stage XYZ stage, 9: laser light source, 10: laser light source XYZ stage, 11: laser light source drive device, 12: microscope, 13: monitor camera, 1
6: Observation part, 17: Cell holding part, 18: Laser light generation part, 19: Control device, 21: Microscope, 22: Electrode, 23: Power supply, 24: Stand, 25: Illumination,
26: Horizontal member of stand, 27: Petri dish, 2
8: Cell, 29: Stage, 30: Interface part, 31: Central processing unit, 32: Storage device, 3
3: output device, 34: input device, 35: data bus, 36: analog / digital converter, 37: command analyzer.

Claims (8)

[Claims] 1. At least one suspension chamber for holding cells to be subjected to fusion, a fusion chamber for fusing cells, a collection chamber for collecting the cells after the fusion treatment, and a field of view. In a cell fusion device having an observation means for observing, a transportation means for transporting cells, an association means for associating cells, a fusion means for fusing cells, and a post-treatment means for performing post-treatment after the fusion treatment, The observation means transmits an image in the field of view as image data to the control device, which has a control device for controlling the operations of the transporting device, the meeting device, the fusing device, and the post-processing device. The control device has a function, at least the function of analyzing the image data, and the conveying unit, the associating unit, the fusing unit, and the post-processing unit according to the analysis result.
And a function of controlling the operation of the means. 2. The function according to claim 1, wherein the function of analyzing the image data has a function of detecting a contour and / or position of a cell and making a judgment based on a shape and / or a position of the contour. Cell fusion device. 3. The transport mechanism using the optical trapping phenomenon according to claim 1, wherein the transport means irradiates cells with light and moves the cells by moving an irradiation position of a spot of the light. And a function of analyzing the image data has a function of detecting an irradiation position and / or a contour of the light spot, and the control device controls the conveyance means based on the irradiation position and / or the contour. A cell fusion device having a function of performing 4. The cell fusion device according to claim 2, wherein the control device has a function of determining the success or failure of fusion based on the contours of the cells. 5. The cell fusion device according to claim 4, wherein the control device controls the fusion means according to the determination of success or failure of the fusion. 6. The associating means according to claim 2, wherein the associating means has an associating mechanism using a dielectrophoresis phenomenon for applying a high-frequency voltage to the cells to associate the cells with each other, and the controller controls the outline of the cells. A cell fusion device having a function of determining the success or failure of an association based on the above. 7. The cell fusion device according to claim 6, wherein the control device controls the association means in accordance with the determination of success or failure of the association. 8. The control device according to claim 2, wherein the control device has a function of selecting a cell to be fused from among the plurality of cells held in the fusion chamber based on the contour of the cell. Cell fusion device.