JPH06253812A - Device for injecting and discharge animal cell or the like - Google Patents

Abstract

PURPOSE:To obtain a device for injecting and discharging animal cells or the like capable of injecting animal cells to a multihole plate useful for cell culture, automatically transplanting to another multi-hole plate and sucking the maximum amount of a medium during exchange of a medium. CONSTITUTION:A device for injecting and discharging animal cells or the like is equipped with a work exchangeable robot of three-axis orthogonal coordinates type controlled by a computer, a finger station 15, a cell tank 17, a reagent station 17, a rinse station 18, a diluter 25 controlled by a computer and an inclinable multi-hole plate cradle 22.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention is a multi-well microwell plate (hereinafter referred to as a multi-well plate) used for culturing animal cells, insect cells, plant cells, etc. and for evaluating and measuring the cultured cells.
The present invention relates to a device for injecting and draining animal cells and the like, which automatically carries out cell transplantation into cells, medium exchange, drug injection and the like.

[0002]

2. Description of the Related Art By culturing cells that retain the tissue function of animals, especially humans, it becomes possible to carry out experiments that reflect human-specific properties. Evaluation of biological activity by test tube experiments and toxicity of drugs Evaluation work has increased significantly under animal use regulations. In this experiment, culture and transplantation of animal cells, medium exchange, drug injection, evaluation and measurement in the evaluation and measurement work, many samples are processed by routine operations. These operations are performed by manually injecting animal cells and medium into a plastic plate or dish having 6 to 96 holes in a biohazard type clean bench by manual pipetting operation and culturing in a CO2 incubator, or when exchanging the medium. Inhalation operation is performed. Therefore, this operation
Due to the large number of treatments, delicate handling of cells, bacterial contamination, etc., it was difficult to automate in a clean bench.

[0003]

Therefore, the present invention is to inject animal cells into a multi-well plate, or to automatically transfer a product that meets the objective conditions to another multi-well plate,
It is another object of the present invention to obtain an automatic injecting / draining device for animal cells or the like that inhales the maximum amount of the medium without damaging the cells at the time of exchanging the medium.

[0004]

SUMMARY OF THE INVENTION The present invention has been made to solve the above problems, and is a computer-controlled work exchangeable three-axis Cartesian coordinate robot, a finger station, a cell tank, and a reagent. Station,
It comprises a rinse station, a computer-controlled dilutor, a multi-hole plate pedestal, and means for inclining the multi-hole plate pedestal.

[0005]

[Function] A probe needle or a tip needle equipped with a finger station is selectively attached to a finger provided at the tip of a Z-axis arm of a computer-controlled three-axis Cartesian coordinate type robot to dilute cells and medium from a cell tank. Is activated to aspirate, and the robot is injected into the well of the multi-well plate having the set well address. Similarly, the injection of the reagent into the well is performed by the robot from the reagent station into the well.
At the time of cell transplantation and medium exchange, the robot and diluter are operated, the multi-well plate pedestal is tilted, and the maximum amount of inhalation is performed to perform cell transplantation and medium exchange.

[0006]

[Examples] As is well known, animal cells are being utilized for drug efficacy screening, toxicity evaluation, monochlor production, etc. in line with the progress of in vitro utilization studies.
Most of these operations are performed manually by hand as described above, but the number of processes required for screening such as cell transplantation, culture, medium exchange, and enzyme activity measurement is enormous, and automation is highly demanded. However, the possibility of automation will be realized by how to functionalize each unique handling according to the floating property and adhesiveness of cells. FIG. 1 shows an embodiment used for transplantation of animal cells into a multi-well plate shown by omitting a part of a piping tube, culture, medium exchange, and color reaction test, and 1 is an animal according to the present invention. The drainage device 3 for injecting cells or the like into the multi-well plate 2 is a well-known multi-well plate reader.

The injecting and draining apparatus 1 according to the present invention comprises a three-axis Cartesian coordinate type robot controlled by a built-in computer, and this robot moves in the Y-axis direction and the arm 4 and in the X-axis direction. The moving body 5 and the lifting body 6 that moves up and down in the Z-axis direction are provided, and the lifting body 6 is provided with a computer-controlled finger 7 as shown in FIG. As shown in FIG. 2, the finger 7 rotates the drive screw 8 in the forward and reverse directions by the motor M provided in the lifting body 6, so that the relay member 9, the pins 10, 10, the bell crank 1
1, 11, pins 12, 12 and finger mounting member 1
It is configured to open and close via 3, 13.

On the base 14, a finger station 15, an animal cell tank 16, a reagent station 17, a rinse station 18, and a tip rack 19 are provided.
, Chip discharge tank 20 and plate lid stocker 21
And a multi-hole plate pedestal 22 is provided. As shown in FIG. 3, the multi-hole plate pedestal 22 is tilted from a horizontal state to 10 ° via a link 24 by an air cylinder 23 operated by a computer instruction. The means for inclining may be a motor-driven cam or crank. Also, the main body 1a
Is provided with a diluter 25 for sucking and injecting animal cells and reagents, a pump, a computer and the like (not shown).

The well-known multi-hole plate reader 3 placed on the base 14 moves the multi-hole plate 2 inside when the multi-hole plate 2 is placed on the loader 31 which appears and disappears. It is possible to identify the wear address of the plate that matches the target condition by measuring the total number of absorbances and performing a comparative measurement with the absorbance level preset in the computer file.

FIG. 5 shows a multiple probe needle 26 and a multiple tip needle 27.
Tip 29 of 7 is a tip rack 1 for every one or several uses
It will be replaced by a new chip at 9 in the robot. Multiple probe needles 26 and multiple tip needles 2
7 is provided with knobs 26a and 27a, respectively, and is sandwiched by the fingers 7 of the robot to be picked up and moved to a desired place.

The use example and operation of this apparatus will be described below. Multi-well plate with animal cells 2
When transplanted to a cell tank 16 containing cells together with the medium
It is necessary to maintain a state in which cells are uniformly dispersed under a constant temperature. For this reason, a device for shaking the cell tank 16 is provided in the container, and the horizontal and horizontal shaking is performed so that cells in a uniformly dispersed state can be moved by a robot that holds the multiple probe needles 26 with the fingers 7. Hole plate 2
Transplant to. The cells transplanted to the multi-well plate 2 are CO2
It is transferred to an incubator and cultured, but the medium replacement work is repeated several times to complete the desired culture.

When this medium is exchanged, the old medium and the new medium are replaced. However, when the old medium is inhaled and discharged, adhered proliferating cells may be damaged or clogged at the tip of the needle. Caliber and inhalation conditions are required. The apparatus of the embodiment shown in FIG. 1 chucks the multiple probe needles 26 in the finger station 15 by a three-axis Cartesian coordinate robot driven by a computer-controlled DC servo motor to remove the cells in the animal cell tank 16 from each other. The sample is sampled by the diluter 25 and dispensed into one row of the multi-well plate 2. This operation is performed for the required number of plates, and the upper lid 21a of the plate 2 is attached to the vacuum pad 2
The workpiece is exchanged for a and each plate 2 sucked from the plate upper lid stocker 21 is subjected to the lid processing.

Next, the multi-well plate 2 is transferred to an incubator (not shown) provided separately, and culturing is performed for a required time. At this time, the medium is replaced if necessary. In case of medium exchange,
The air cylinder 23 controlled by the computer is operated to tilt the angle of the multi-hole plate 2 upward from the horizontal position as shown in FIG. This angle range is preferably adjusted according to the size of the multi-hole plate 2 and is tilted up to an angle of 10 °. The sampling position in each well of the multi-well plate 2 is determined by the push-up distance proportional to the tilt angle, the inter-well distance, and the number of wells.

To each of the proliferated and transplanted cells, a drug or a coloring reagent is added by a robot from a reagent station 17 with a multiple probe needle to search for biological activity of a receptor or a ligand substance, and the generation state is monitored. Therefore, the multi-hole plate 2 is attached to the loader 3 of the plate reader 3.
Move to 1 and measure the total absorbance of the multi-well plate, compare it with the absorbance level set in the computer file in advance, identify the well address of the multi-well plate that matches the target condition, and specify the well The solution inside is sampled and re-implanted into a new multi-well plate. If the colored multi-well plate is not available, the specified multi-well plate is used as the original plate by duplicating the same sample before color development, and the well of the corresponding specific address Use solution.

When the multiple chips 27 are used, the used chips are removed in the chip discharge tank 20 and discarded. Further, when washing the multiple probe needles 26, the multiple probe needles are inserted into the rinse station 20 and the diluter 25 is operated to suck and discharge the washing liquid in the rinse station 20 for washing. As described above, for convenience of explanation, the explanation has been made in relation to animal cells, but it goes without saying that the device of the present invention can be used not only for animal cells but also for insect cells and plant cells.

[0016]

INDUSTRIAL APPLICABILITY According to the present invention, transplantation of animal cells, insect cells, plant cells and the like to the multi-well plate 2 and transplantation of cultured cells in the multi-well plate 2 to the multi-well plate are automatically performed. It can be performed, and the efficiency of cell culture work can be improved. Further, it is possible to inhale the maximum amount of the medium without damaging the cells at the time of changing the medium.

[Brief description of drawings]

FIG. 1 is a perspective view of an embodiment of the present invention.

FIG. 2 is an explanatory diagram showing a finger driving mechanism in the embodiment.

FIG. 3 is a plan view of a finger in the example.

FIG. 4 is an explanatory view of a tilting mechanism in the embodiment.

FIG. 5 is a rear view of the multiple probe needle and the multiple tip needle in the example.

FIG. 6 is a view showing a state of inserting a needle into a multi-well plate at the time of cell injection and medium exchange.

[Explanation of symbols]

 1 Injection / Drainage Device 2 Multiwell Plate 3 Multiwell Plate Reader 4 Arm 5 Moving Body 6 Lifting Body 7 Finger 8 Drive Screw Rod 9 Relay Member 10 Pin 11 Bell Crank 12 Pin 13 Finger Mounting Member 14 Base 15 Finger Station 16 Cell Tank 17 Reagent Station 18 Rinse Station 19 Chip Rack 20 Chip Discharge Tank 21 Plate Top Lid Stocker 22 Multi-hole Plate Cradle 23 Air Cylinder 24 Link 25 Dilutor 26 Multiple Probe Needle 27 Multiple Tip Needle 28 Exchange Tip 29 Vacuum Pad

Claims (1)

[Claims] 1. A 3-axis Cartesian coordinate robot capable of computer-controlled work exchange, a finger station, a cell tank, a reagent station, a rinse station, a computer-controlled dilutor, and a multi-well plate pedestal. And a means for inclining the multi-well plate pedestal, an apparatus for injecting and draining animal cells and the like.