JPS6260061B2 - - Google Patents

Description

Detailed Description of the Invention [Industrial Application Field] The present invention provides a cell sorting device that can efficiently select target cells from a group of cells or a group of microorganisms containing several types of cells or microorganisms. It is related to.

[Conventional technology]

Monoclonal antibodies are useful for research on molecular structure, genes, and their functions in antibody chemistry and antigen chemistry.
It is used in the study of receptors for hormones and neurotransmitters in pharmacology, as well as in virology, parasitology, and bacteriology. Also, in diagnosis,
It is also used for molecular biochemical analysis of immunodeficiency diseases and detection of antigens (malignant tumors, etc.).In addition, it is also used in the treatment of organ transplantation (histocompatibility), passive immunity (injection of antibodies), and detection of malignant tumors. It is used in the treatment of tumors.

As mentioned above, monoclonal antibodies have a wide range of applications.

Secretions such as monoclonal antibodies are obtained from cells that produce them, and conventional methods for selecting and producing monoclonal antibody-producing cells include, for example, the following methods.

Splenocytes (2.5×10 ⁸ cells), tumor cells (2.5×10 ⁷ cells)
Cell fusion was carried out using polyethylene glycol as a fusion promoter, and after centrifugation, the fusion solution was discarded and HAT culture solution (Hypoxanthine Aminopferin and Thymidine
Add culture solution (containing culture solution) to disperse the cells. Thereafter, the cell suspension obtained by dispersion is dispensed, for example, 0.2 ml each into a plurality of wells provided in the tray, and then cultured in a CO ₂ incubator for two weeks. This culture kills unfused cells and proliferates fused cells.

Whether or not fused cells are proliferating is observed by the presence or absence of colonies, and wells in which fused cells are found to be proliferating are tested with ELISA (Enzyme Inked immunosorbent).
assay) Collect and dispense the culture supernatant into a detection tray. Thereafter, whether or not the antibody of interest is contained in the collected culture supernatant is detected by ELISA. At this time, out of the 480 wells to which the cell fluid is dispensed, usually only about 5 wells produce the antibody of interest.

into the cell fluid of the well producing the desired antibody.
Add diluent to the extent that the cells can be counted using a microscope to uniformly disperse the cells. (Initial dilution). Next, a portion of the initially diluted cell fluid is counted on a blood count plate and observed under a microscope to calculate the total number of cells contained in the cell fluid.

Based on the calculated number of cells, dilute the cell solution so that when 0.2 ml is dispensed into each well, one cell is dispensed into every three wells (cell solution dilution). This cell solution dilution is to increase the probability that two or more cells will not be dispensed into one well in order to obtain a single clone.

Thereafter, in order for the cells to proliferate, it is necessary to have a certain number of cells, so thymocytes that do not have the ability to produce antibodies are added.

Repeat the above operations (adding HAT culture solution, dispersing the cells, dispensing the cell solution into wells, and adding thymocytes; this series of operations is called limiting dilution) twice. , the probability of monoclonality is increased, the obtained monoclonal antibody-producing cells are cultured in large quantities, and monoclonal antibodies are produced in large quantities.

[Problem that the invention seeks to solve]

Conventionally, as mentioned above, monoclonal antibody-producing cells have been manually selected from a group of cells that have undergone cell fusion, but the probability of obtaining the desired monoclonal antibody-producing cells is low. However, there are problems such as low stability of the obtained cells (that is, they are likely to die) and the risk of contamination with bacteria during the selection process, and the production of large amounts of monoclonal antibodies with high activity. When trying to select cells, it is necessary to select from a vast number of fused cells.

Furthermore, this sorting work requires a high level of skill, and it usually takes 1-2 hours to acquire this skill.
Because this method requires years of training, there are very few technicians who can select monoclonal antibody-producing cells.
For this reason, the number of cells that can be handled in a series of experiments is limited, and the probability of obtaining the desired monoclonal antibody-producing cells is low, and even if they are obtained, the result is only a low antibody-producing ability. This has become an inhibiting factor in the application of monoclonal antibodies.

[Means for solving problems]

The present invention has been made to solve the above-mentioned problems, and its gist includes a preparative dispensing section, a tray transport section, a culturing section, an assay section, a cell counting section, an isolation dispensing section, and The cell sorting device includes a control, and the sorting and dispensing unit includes a pipette, a manipulator that grips the pipette, and a pump for aspirating and discharging a fixed amount of liquid with the pipette, and the tray transport unit includes a pipette and a manipulator that grips the pipette. It consists of a tray stand to be placed on and a drive means for transporting the tray stand to a predetermined position in the preparative dispensing section, the culturing section, the assay section, and the isolation dispensing section, and the culturing section has a tray stock for storing a plurality of the trays. Equipped with
The cell number counting section includes a microscope, a drive motor that moves the preparation to a position under the microscope, and a control circuit that counts the number of observed cells, and the isolation and dispensing section includes a dilution section consisting of a dilution container and the dilution section. The controller moves the tip of the pipette into the cell solution container and onto the well provided in the tray on the tray stand of the transport section. means for transmitting a signal to the manipulator to cause the pipette to inject and discharge a solution; means for transmitting a signal to the pump for injecting and discharging a solution into the pipette; means for transmitting a signal to the drive means of the transport section; means for transmitting a signal to the culture section tray stand for moving the tray between the culture section tray stand and the tray stock; and a means for transmitting a signal to the manipulator to move onto the wells in the detection tray, and a means for transmitting a signal to the pump to inject the supernatant of the wells in the tray after culture into a pipette and discharge it to the wells in the detection tray. means for transmitting a signal to the tray conveyance section to carry the detection tray into the assay section; and means for transmitting a signal to the pipette manipulator for moving the tip of the pipette into a well provided in the tray and onto the preparation. a means for transmitting a signal to the pump to cause the pipette to inject the cell solution in the wells in the tray after culturing and discharge it onto the preparation; and a means for transmitting a signal to the pump to cause the cell solution to be moved to an observation position under a microscope in the cell counting section. When the number of cells counted by the control circuit is above a certain value, the tip of the pipette is inserted into a well containing the same solution as the cell solution observed under the microscope. means for transmitting a signal to the manipulator to move the cell onto the dilution container; means for transmitting a signal to the dilution section for calculating the necessary dilution amount based on the counted number of cells and adding the calculated amount of diluent to the dilution container; A means for transmitting a signal to the cell isolation unit to introduce the solution in the container into the capillary tube, and a drive unit for the tray transport unit that synchronizes the detection signal of cells passing through the capillary tube and sends a signal to move the tray stand at well intervals. The present invention resides in a cell isolation device characterized in that it is a device comprising means for transmitting a signal, and is a device that controls each of the units according to input operating conditions of each unit.

[Action of the invention]

The operation of the device of the present invention will be explained below.

The solution containing the fused cell group is dispensed in fixed amounts into wells (solution reservoirs) on the culture tray using the preparative dispensing unit, and the culture tray is then transferred to a tray stock that can store multiple trays using the tray transport unit. Transfer to the equipped CO ₂ incubation unit. Culture trays that have been stored in the CO ₂ culture section for a certain period of time under conditions that allow the target cells to proliferate are returned to the tray transport section, and if it is necessary to detect proliferation by colony observation, a colony observation section is provided. Detect whether colonies are growing in each well. A predetermined amount of the supernatant of the well in which colony growth has been detected is dispensed into a detection tray by an automatic sorting/dispensing section.
The detection tray filled with supernatant liquid is transported to the assay section by a tray transport mechanism, and it is detected whether the supernatant liquid contains antibodies. As a result,
For example, if the cell concentration in the solution is unknown or inconsistent, a portion of the culture tray wells that have been confirmed to contain antibodies in the supernatant may be automatically sorted. Transfer the solution to the cell counting section using the injection section, and measure the cell concentration in the solution. If the cell concentration is higher than that at which it is possible to separate and dispense cells one by one, the dilution mechanism is used to dilute the cell to the desired concentration, and the diluted solution is then added to a new cell via the isolation and dispensing section. Dispense each cell into each well of the culture tray. This culture tray, in which cells are dispensed one by one into each well, is prepared by adding other cells if necessary for proliferation.
Monoclonal antibody-producing cells are obtained by storing the cells in a CO ₂ culture chamber and repeating the above-described mechanism once again until each cell is dispensed from there. These operations are automatically controlled by the controller according to conditions input from the outside. Of the above operating mechanisms, if there is no need to measure the degree of cell proliferation, or
In cases where the number of proliferated cells is suitable for normal operation of the isolation and dispensing mechanism, the colony observation section and dilution mechanism may not be particularly provided.

〔Example〕

Embodiments of the present invention will be described along with their functions with reference to the drawings. FIG. 1 is a perspective view showing the cell sorting device of the present invention.

A container 1, such as a test tube, containing fused cells and a culture solution is grasped by a container handler 2, and the container 1 is moved to a location of a pipette manipulator 4. A chuck operated by air drive or motor drive is attached to the tip of the container handler 2, and grips these containers 1. The pipette 3 is inserted into the container 1, and the cell solution is aspirated using the pump 5. The pipette manipulator 4 is moved to the well position of the culture tray 18, and the pump 5 discharges a certain amount of cell solution into the well. The pipette 3 is moved onto another well by the movement of the tray transport section 16 or the pipette manipulator 4, and a certain amount of cell solution is similarly discharged into the well. These operations are repeated to dispense the cell solution into each well on the culture tray 18. The pipette manipulator 4 can use a horizontal multi-joint type arm, orthogonal type arm, etc., and the operation of the tray conveyance unit 16 can be controlled by a motor drive or a third type arm as shown in FIG.
A driving means such as an air drive as shown in the figure can be used. In addition, in Fig. 2, 39 is a feed screw (ball screw), 40 is a pulse motor,
In FIG. 3, 41 is an air cylinder, 42 is an electromagnetic brake, 43 is a potentiometer, and 44 is an electromagnetic valve. Other wire drives can be used. The culture tray 18 into which the solution has been dispensed is transferred to the CO ₂ incubator 24 via the tray stand 19.
The cells are stored in a tray stock 20 inside the cell and maintained in a cell culture environment for a certain period of time. tray stock 20
In order to store the culture tray 18 in the tray, first, the tray conveyor 16 moves the culture tray 18 to a position where it contacts the tray stand 19.
By moving the tray chuck 17' on the tray chuck 9, the culture tray 18 is stored in the tray stock 20 through the tray stand 19. The tray chucks 17 and 17' can be opened and closed in the θ direction and moved in the x and z directions by driving a motor as shown in FIG. 4, for example. In addition, in Fig. 4, 45 is a motor;
6 is a ball screw, and 47 is a peon rack.
Alternatively, these operations can also be performed by air drive. The culture tray 18, which has been kept in the cell culture environment in the CO ₂ incubator 24 for a certain period of time, is moved to the tray transport section 16 by the reverse operation of inserting it into the tray stock 20, and is transferred to the colony observation section 26.
Each well on the culture tray 18 is moved to the position shown in FIG. 1, and it is detected whether colonies are growing. The colony observation section 26 is composed of an image input section consisting of a TV camera with a microscope and an image processing section consisting of a computer, and performs digital image processing on the enlarged image taken by the input section, thereby observing the colony. Perform detection. The image processing method uses the high brightness of living cells to detect living cells using a binarization method, and it is possible to detect colonies from the proportion of living cells in the entire screen. . Alternatively, colonies can be detected by detecting the amount of light transmitted. "Tray 1
If no colonies are detected in all wells of 8, pipette 3 moved by pipette manipulator 4 is used to remove the supernatant of the cell solution, that is, the old culture solution. suction,
Furthermore, the pipette manipulator 4 is moved and the sucked culture solution is discarded into the drainage tank 53. Next, in the same manner, the culture solution is replaced by moving the pipette manipulator, sucking a new culture solution from the culture solution tank 54, and discharging it into the well from which the old culture solution was sucked. The above operations are performed for all the wells during culture, and the tray 18 is carried into the tray stock 20 in the CO ₂ incubator 24 again by the operations described above, and culture is continued for a certain period of time.

On the other hand, if a colony is detected, the culture tray 1 on which the colony was detected to be growing
From each well on 8, pipette manipulator 4
The supernatant of the cell solution is aspirated by the pipette 3 moved by the pipette 3, and a fixed amount is dispensed into each well of the detection tray 9. Do this for all wells. In addition, pipette manipulator 4
Add a fixed amount of the assay reagent 7 to the wells on the detection tray 9 containing the cell solution supernatant using a holder, and move it into the assay section 8 by motor drive or air drive in the same way as the tray transport section 16. ,
It is detected whether the target substance is contained in the cell solution supernatant placed in each well of the detection tray 9. The assay section uses liquid chromatographs and ELISA (Enzyme Inked).
immno sorbent assay) analyzer, etc. can be used. Detection that the target substance is contained in the cell solution supernatant means that
This means that the cells in the wells of the original culture tray 18 are producing the target substance. A portion of the cell solution that produces the maximum amount of the target substance in the wells of the culture tray 18 that is producing the target substance is transferred onto the preparation of the preparation transfer unit 31 using a pipette manipulator at 30°. The cell solution is dispensed, and the number of cells contained in the cell solution on the preparation is counted by the cell number counting section 25. The pipette manipulator 30 is the same as the pipette manipulator 4, and has a pipette 31 at the tip and
A peristaltic pump 28 connected to this is attached. The preparation transport section 32 is a linear movement mechanism composed of a drive motor and a feed screw,
Preparation 55 is placed on the moving stage in advance.
is placed. The cell counting section is equipped with a microscope.
It is composed of an image input section 25 consisting of a TV camera and an image processing section 27 which is a computer, and measures the number of living cells by digital processing similarly to the colony observation section 26. A dilution rate for calculating the cell concentration of the cell solution in the well on the original culture tray 18 based on the number of cells counted by the cell number counting section 25 and diluting it to the desired concentration that allows isolation and dispensing. Determine. Next, the pipette manipulator 30
Dilute a certain amount of the cell solution in the well with the container 33
A predetermined amount of diluent is added by the diluting section 34 to dilute the solution to a desired concentration. Cell concentration is calculated by the following method. That is, the amount of cell solution in the well and the amount of sampling on the preparation 55 are kept constant, and the amount is calculated by back calculating the volume ratio from the number of cells on the preparation. Dilution section 34
consists of a diluent tank and a pump, and controls the amount of diluent discharged by controlling the pump time.

The diluted cell solution is inhaled by the isolation/dispensing section 35. In the isolation/dispensing section, a diluted electrolyte flows through a thin tube, and the resistance of the electrolyte flowing between the electrodes is different depending on whether there are cells or not, which is used to determine the current value. Detect cells by changes. The mechanism of the isolation/dispensing section is, for example, as shown in FIG. Cells can be detected by receiving a light-emitting part and a light-receiving part so as to sandwich a thin tube instead of using electrodes, and by sensing the interruption of light from the light-emitting part by the passage of cells with the light-receiving part.

The isolation and dispensing section 35 is composed of a suction pump 56 and the above cell detection sections 48 and 49, and when no cells are detected, the diluted solution is discarded, and when cells are detected, this cell detection signal is The culture tray 51 is moved by the tray transport unit 16 in synchronization with the culture tray 51, and cells are dispensed one by one into each well in the culture tray 51 that is not used for culture.

This is the liquid feed pipe 5 from the cell detection section to the well.
8 is set to a value smaller than the volume of the well, and when a cell detection signal is obtained, the suction pump of the isolation/dispensing section 35 is temporarily stopped. Then, after driving the tray transport unit 16 and moving the well into which cells are to be dispensed next to directly below the liquid feeding tube, the suction pump is restarted and the cells are dispensed into the well. In this case, if two or more cells exist in the liquid delivery tube from the cell detection unit to the well, one cell cannot be dispensed into one well. , dilute to the required and sufficient value. This value is determined experimentally in advance. The culture tray into which each cell has been dispensed is stored again in the tray stock 20 within the CO ₂ incubator 24 to culture the cells.

Furthermore, in the same manner as described above, colony detection and target substance detection are performed, and target cells are selected by the above process.

The controller 36 is a device comprising, for example, an input device 37 and a control section 38, and controls each section according to the above-mentioned sections and operating conditions between the sections inputted through the input device.

A block diagram and an operational flow diagram showing the signal system in the present invention are shown in FIGS. 6 and 7, respectively.

In FIGS. 6 and 7, the control signal S ₁ sent from the controller is a signal for controlling the opening and closing of the chuck in the container handler 2, the rotation and raising and lowering of the container handler 2, and the control signal S ₂ is a signal for driving the manipulator 4. This is a signal that controls the motor. Further, the control signal _S3 is a signal for operating the pump 5.

The signals S ₁ , S ₂ and S ₃ actuate the container handler 2 and the manipulator 4 to remove the pipette 3 from the container 1.
The cell solution is separated by .

Next, the manipulator 4 is actuated by the signal _S2 , and the pipette 3 is moved to a position above one of the plurality of wells provided on the tray 18.

The control signal _S3 activates the pump 5 and pours the cell solution contained in the pipette 3 into the well.

Next, the stepping motor 10 in the tray conveying section 16 is operated by the control signal _S5 to move the tray 18 a distance corresponding to one well on the tray, and then the above operation of the manipulator 4 is repeated.

After dispensing the cell solution to all wells on the tray 18, the tray chuck 17 in the tray transport section 16 is operated (opening/closing in the θ direction in FIG. 4).
the signal S ₄ for controlling the movement in the x and y directions, the aforementioned control signal S ₅ , and the tray chuck 1 in the culture section.
A signal S ₆ for controlling the operation of the drive motor 7' and a signal S ₇ for controlling the operation of the drive motor 21 in the incubator 24 are transmitted from the controller 36 to each part, and the tray 18 is moved to the CO ₂ incubator 24 and placed in the tray stock 20. It will be stored.

In the culture section, the culture conditions include opening/closing the door of the CO ₂ incubator 24, culture temperature, culture time, and CO ₂
A control signal S ₈ containing the amount (opening/closing of a valve installed in the CO ₂ supply pipe) and humidity (replenishment of water to a water tray installed in the incubator, opening/closing of a steam valve, etc.) set and maintained.

After the cultivation is completed, the tray 18 is moved to the colony observation section by reverse operation using control signals S ₄ , S ₅ , S ₆ , and S ₇ .

In the colony observation section, the image of the colony observed by the microscope-equipped TV camera 26 is used as a measurement signal.
The measurement signal T ₁ is sent to the controller 36 as T 1 , and the controller 36 performs image processing on the measurement signal T ₁ .
Calculate the percentage of live cells in the entire colony screen.

Next, the manipulator 4 is actuated by the signal _S2 , and the pipette 3 is moved to a position above one of the plurality of wells provided on the tray 18 in which the degree of cell proliferation has been determined.

If no colonies are detected in all the wells of the tray 18, the pump 5 is activated by the control signal _S3 , a certain amount of the culture supernatant from each well is taken out by the pipette 3, and the control signal is again activated. Signal S ₂
The culture supernatant in the pipette 3 is then moved to the corresponding wells in the antibody detection tray 9, and the culture supernatant in the pipette 3 is dispensed into each well of the antibody detection tray 9 in accordance with the control signal _S2 .

According to the control signals _S2 and _S3 , the pipette is changed every time the cell culture supernatant liquid is dispensed into each well of the antibody detection tray.

When the culture solution in all wells on the culture tray is replaced, the culture is continued by the control signals S ₄ to _{S 8} described above.

On the other hand, if a colony is detected, the control signal S ₂
Move pipette 3 to the assay reagent position by
and aspirate the assay reagent into the pipette ₃ using the control signal S3. Next, the assay reagent is discharged into each well of the detection tray 9.

Next, in accordance with the control signal _S9 , the antibody detection tray 9 placed on the upper part of the tray stand enters the assay section by actuation of the driving means. The measurement signal _T2 contains the amount of antibody contained in the cell solution supernatant measured in the assay section, and the controller calculates the antibody production ability of the cells (=antibody amount measurement value/proliferation degree measurement value).

In response to the control signal S ₅ , the culture tray 18 is moved to the position of the pipette manipulator 30 by the tray transport section, and then the manipulator 30 is actuated in response to the control signal S ₁₁ to move the pipette 4 to the preparation plate 55 .
The cell solution in the pipette 31 is dropped onto the preparation 55 by operating the pump 55 using the control signal _S12 .

The preparation 55 is moved below the microscope 25 using the control signal _S10 , and the cell number counting section counts the number of cells in the cell solution. measurement signal
_T3 is the number of cells, and its value is used in the controller to calculate the cell concentration in the cell solution. Control signal S ₁₁ when cell concentration is above a certain value
The pipette of the manipulator 30 is moved to the well containing the cell solution having the cell concentration, and the pump is activated by the control signal _S12 to dispense the cell solution into the pipette. Next, the pipette that has dispensed the cell solution is moved to the dilution container by the control signal _S11 , the cell solution is poured into the dilution container by the control signal _S12 , and the diluted solution is poured into the dilution container by the control signal _S13 . and dilute it below a certain concentration.

The diluted cell solution is sucked into the cell isolation separation unit 35, and when the cells 48 pass through the thin tube 57,
Emit measurement signal T ₄ .

The control signal is synchronized with this cell detection measurement signal.
S ₃ and S ₄ actuate the tray conveyance section, and the liquid sending pipe 58
The distance between the wells on the tray is moved at the exit 59 of the tray.

Through the above-described process using the cell sorting device of the present invention, individual cells can be isolated from a group of cells.

〔Effect of the invention〕

Since the cell sorting device of the present invention has the above-described configuration, it has the following effects.

(b) Selection of monoclonal antibody-producing cells can be easily carried out without special advanced technology by simply setting the conditions for each mechanism and starting the operation of the device, which greatly reduces labor. can be reduced to

(b) There is no risk of contamination with bacteria and the amount of processing can be increased, so monoclonal antibody-producing cells can always be stably obtained and efficiency can be increased.

(c) Conventionally, it took two to three months to obtain one monoclonal antibody-producing cell, but with this device, it can be obtained in about one month, including the proliferation of monoclonal antibody-producing cells. During this time, several types of single clones can be processed continuously and simultaneously.

[Brief explanation of the drawing]

Figure 1 is a perspective view of the cell sorting device of the present invention, Figure 2 is a perspective view of the cell sorting device of the present invention;
The figure is a side view schematically showing an example of a tray transport mechanism that is driven by a motor, FIG. 3 is a side view schematically showing another example of a tray transport mechanism that is air driven, and FIG. 4 is a side view of a tray transport mechanism that is driven by an air drive. FIG. 5 is a diagram showing a tray stand, a tray chuck, and a motor drive as an example of driving the tray chuck, in which A is a plan view thereof, B is a side view thereof, and FIG. 5 is a schematic diagram of an isolation and dispensing section. FIG. 6 is a block diagram of the present invention, and FIGS. 7A, B, C, D, and H are operational flow diagrams of the present invention. DESCRIPTION OF SYMBOLS 1... Container, 2... Container handler, 3... Pipette, 4... Pipette manipulator, 5... Pump, 6... Pipette exchange section, 7... Assay reagent, 8... Assay section, 9 ...Detection tray, 1
0... Drive motor, 11... Drive motor, 12
...Feed screw, 13...Feed screw, 14...Guide, 15...Guide, 16...Tray conveyance section, 1
7... Tray chuck, 18... Culture tray,
19...Tray stand, 20...Tray stock, 2
1... Drive motor, 22... Feed screw, 23...
Guide, 24... CO ₂ incubator, 25...
Cell number counting section, 26...Colony observation section, 27
...Control unit, 28...Pump, 29...Drive motor, 30...Pipette manipulator, 31...
Pipette, 32...Preparation transport section, 33...
... dilution container, 34 ... dilution section, 35 ... isolation and dispensing section, 36 ... controller, 37 ... input device,
38... Control unit, 39... Feed screw, 40... Pulse motor, 41... Air cylinder, 42... Electromagnetic brake, 43... Potentiometer, 44...
... Solenoid valve, 45 ... Motor, 46 ... Ball screw, 47 ... Peon rack, 48 ... Cell,
49... Electrode, 50... Ammeter, 51... Well.

Claims (1)

[Claims] 1 A cell sorting device consisting of a preparative dispensing section, a tray transport section, a culturing section, an assay section, a cell counting section, an isolation dispensing section, and a controller, wherein the preparative dispensing section includes a pipette and a pipette. It consists of a manipulator for gripping and a pump for suctioning and discharging a fixed amount of liquid with the pipette, and the tray transport section includes a tray stand on which the tray is placed, and a tray stand that connects the tray stand to the preparative dispensing section, culture section, assay section, and unit. It consists of a driving means for transporting the preparations to a predetermined position in the separation/dispensing section, the culture section includes a tray stock for storing a plurality of the trays, and the cell counting section moves the preparations to a microscope and a position under the microscope. The isolation and dispensing section consists of a drive motor and a control circuit for counting the number of observed cells, and the isolation and dispensing section includes a dilution section consisting of a dilution container, a thin tube through which the cell solution passes through the dilution section, and a cell that detects cells passing through the thin tube. The controller includes means for transmitting a signal to the manipulator to move the tip of the pipette into the cell solution container and onto the well provided in the tray on the tray table of the transport section, and a means for injecting and discharging the solution into the pipette. means for transmitting a signal to the pump to move the tray carrying tray to the front of the culture tray tray; a means for transmitting a signal to the culture section tray stand to move between the tray stocks; and a means for transmitting a signal to the manipulator to move the tip of the pipette into the supernatant of the well in the tray after culture and onto the well in the detection tray. means for transmitting a signal to the pump for injecting the supernatant liquid from the wells in the tray after culture into a pipette and discharging it into the wells in the detection tray; and a means for transmitting a signal for transporting the detection tray to the assay section. means for transmitting a signal to the pipette manipulator to move the tip of the pipette into the wells provided in the tray and onto the preparation; means for transmitting a signal to the pump to cause the sample to be ejected onto the slide; means for transmitting a signal to the drive motor of the slide transfer unit for moving the sample to an observation position under a microscope in the cell counting unit; and a control circuit. means for transmitting a signal to the manipulator to move the tip of the pipette into a well containing the same solution as the cell solution observed by the microscope and onto the dilution container when the number of cells counted by the cell count is above a certain value; A means for calculating the required dilution amount based on the number of cells obtained, and transmitting a signal to the dilution section to add the calculated amount of diluent to the dilution container, and a means for transmitting a signal to the cell isolation section for introducing the solution in the dilution container into the capillary. A device comprising a means for transmitting a signal, and a means for transmitting a signal for moving the tray stand at well intervals to the drive means of the tray conveying section in synchronization with the detection signal of cells passing through the capillary, A cell isolation device, characterized in that it is a device that controls each of the above-mentioned parts according to operating conditions.