JPS5953023B2 - Pipette feeding device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a pipette feeding device for feeding a pipette into a culture chamber in an apparatus for automatically culturing living tissues and cells.

In all fields such as medicine, biology, pharmacy, and agriculture,
Biological tissue and cell culture techniques are essential basic experimental techniques for conducting research at the cellular level.

However, due to the technical difficulties in subculturing biological tissues and cells, stable culture strains could not be obtained. However, recently, with the spread of gas culture technology in incubators, that is, culture technology in a specific gas atmosphere, special cells that were previously considered difficult to grow, such as those of the liver, nervous system, and pituitary gland, have become more popular. Even subculture is becoming possible.

The main points of such cell-based methods currently in use are described below.

First, in order to perform subculture, a predetermined number of cells are diluted with a culture medium in a culture container such as a Petri dish, and cultured stationary in an incubator maintained in a predetermined atmosphere. To check the culture status after the passage of time, take out the specified culture container from the incubator and examine the degree of cell proliferation using a microscope.This will confirm that the desired cells have proliferated to fill the specified container. Once it is confirmed that the cells are present, transfer them to sterile clean pliers, aspirate and discard the culture solution in the culture container with a pipette, and wash the remaining cells in the container by injecting a buffer solution. The buffer used for this washing is aspirated again and discarded.Next, in order to release the cells that have settled and grown on the bottom of the culture vessel from the culture vessel, an enzyme such as trypsin is injected, and the cells are left for several minutes. Then, discard this. After leaving it for a few more minutes, inject the culture solution. This culture solution has the effect of stopping the action of trypsin. After injecting the culture solution, stir it and transfer it to a centrifuge tube. , perform centrifugation. Aspirate and discard the supernatant of this centrifuged product. Next, inject the culture solution and stir to resuspend the cells, and transfer a fixed amount of the resuspended cell suspension to a new culture container. After the dilution and dispensing operation has been completed, the culture container is taken out using clean pliers and placed into an incubator maintained at a predetermined atmosphere. Transfer, leave to stand, and proceed with culturing again.5 However, the method described above has the following drawbacks.

One of them is that in order to check the growth state of tissues or cells using a microscope, it is necessary to frequently remove the culture container from the incubator to the outside air.

Therefore, when the culture condition is taken out from the predetermined environmental conditions such as gas atmosphere, temperature, humidity, etc. into the outside air, the culture conditions may change suddenly, and subtle changes may occur in the cultured tissue or cells. Also, since it comes in contact with the outside air, it is susceptible to contamination by bacteria. In this way, it is directly affected by changes in environmental conditions and the contamination of various bacteria. Next, based on the results of microscopic observation, the technician manually performs the above-mentioned subculture operation in a clean bench, so there is a direct effect on tissues or cells during the subculture operation of the technician. .

In other words, this shows that it is not possible to perform standardized culture under certain conditions with such conventional subculture operations. The tissue or cells cultured will also depend on the experience and skill of the technician. This means that it is difficult to standardize and unify the culture technology itself.For this reason, even if researchers conduct research on the same theme, researchers may come to completely opposite conclusions. It is often seen and heard. Furthermore, it takes a considerable amount of time to train technicians equipped with these culture techniques, and the absolute number of tools is insufficient, making it difficult for researchers to concentrate on their original research. The reality is that a considerable amount of energy must be spent on technology.

The goal is to prevent contamination of outside air caused by the conventional 4-in-1 method, eliminate the influence of human operations, and standardize and unify each operation in subculture. Automatic culture devices are being developed for the purpose of automatically culturing standardized tissues or cells. This automatic culture device is capable of transferring the aforementioned proliferated cells from the culture container to the centrifuge, discarding the supernatant after centrifugation, and injecting the culture solution into the centrifuge and resuspending the cells. A dispensing device for dispensing to a new culture container, etc. is installed in a culture chamber maintained at a constant atmosphere, and the above-mentioned operations are performed in this culture atmosphere. Furthermore, during these operations, for example, the pipette used to discard the supernatant after centrifugation becomes contaminated with the supernatant, so it must not be used for injecting the next culture solution or transferring it to a new culture vessel. It is not preferable to dispense several cells. Therefore, the above-mentioned problem of contamination can be solved by making the pipette easily attachable and detachable so that the pipette can be replaced each time it is used.

For example, the upper part of the inner surface of a pipette is tapered, and the pipette is held and used by fitting it into a holding member whose tip end is also tapered, and after use, it is removed from the holding member and discarded. Furthermore, a new pipette is attached and used, and after use, it is removed from the holding member and discarded. However, in this method, when the pipette is attached to the holding member, the pipette is installed in a culture chamber that maintains a constant atmosphere, that is, a culture atmosphere, so the culture atmosphere mentioned above comes into direct contact with the outside air. This may cause the culture atmosphere to mix with outside air, changing the indoor atmosphere and introducing bacteria.

Furthermore, although the pipette that is attached is sterilized, if the pipette is brought into the culture chamber manually, there is a risk that the pipette, especially the immersed part at the tip, may come into contact with outside objects and become contaminated. . The present invention has been made in view of the above points, and provides a pipette feeding device that moves a pipette from one atmosphere to the other between two different atmospheres without the two atmospheres coming into direct contact. This is what we provide.

The specific contents of the pipette feeding device of the present invention will be explained below based on the drawings. In FIG. Although not shown, the pipette holding arm is movable and is located in a chamber 4 maintained in a culture atmosphere together with other devices of the pipette.

Next, the specific structure of the pipette feeding device 2 will be explained. Reference numeral 11 denotes an opening (export (1)
12 is a shaft fixed to the center of the main body 11, and 13 is a cylindrical rotating member having a notch 13a. , are installed in the main body 11 so as to be rotatable about an axis 12.

Reference numeral 14 denotes a plate-shaped member having a guide groove 14a, which is fixed to the shaft 12. Reference numeral 15 indicates a plate-shaped member having a guide groove 14a, and is provided to push the pipette and move it along the guide groove.
A rotary plate fixed to this so as to rotate together with 3,
It has a notch 15a in which the upper part of the pipette is placed.

16 is an opening for feeding the pipette (carrying (1) 16
It is a lid of the main body 11 having a.

Next, the operation of the pipette feeding device of the present invention will be described. In FIG. At this time, the pipette 10 is dropped.

As a result, the pipette 10 is inserted through the opening 16a,
The tip of the guide plate 14 is inserted into the notch 13a of the rotating member 13 from the guide groove 14a of the guide plate 14. At this time, if the width of the guide groove 14a is made wider than the lower part 10a of the pipette 10 and narrower than the upper part 10b, then
The pipette 10 is held by a guide plate 14 as shown in the drawings. Here, the rotating member 13 and the rotating plate 1 are connected by appropriate means.
5 are rotated together clockwise in FIG. 2, the rotary plate 15 pushes the pipette with its notch 15a and moves the pipette. This movement of the rotating plate 15 causes the pipette to move along the guide groove 14a of the guide plate 14. In this way, when the rotating member 13 and the rotating plate 15 rotate together and move from the position shown in A to B in FIG. 2, the opening 11a formed in the main body 11
In the position shown in FIG. 2A, what was previously closed by the rotating member is completely opened. At this time, the pipette holding arm 3 shown in FIG. 1 is rotated and its tip enters through the opening 11a as shown by the chain line in FIG.
4b. In this state, if the rotating member 13 and the rotating plate 15 are further rotated, the position shown in FIG. 3b, and the pipette is held in the holding arm. Now, when the pipette holding arm 3 is rotated again, the pipette is thereby moved and transferred to a pipette holding device (not shown). During the operation described above, first, the rotating member 13 is placed in the position shown in FIG.
When the rotating plate 15 is in position, the opening 16a of the lid 16 is open, but the opening 11a of the side wall of the main body 11 is completely closed by the rotating member 13. When the rotating member 13 and the rotating plate] 5 rotate together, the rotating plate 15 first moves to the lid 16.
The rotating member 13 also closes the opening 16a of the lid 16. In this way, after the opening 16a is completely closed, the opening 11a of the main body 11 is opened as shown in FIG. 2B. Further, as described above, after the holding arm 3 rotates with the pipette and moves from the vicinity of the opening 11a of the main body 11, the rotating member 13 and the rotating plate 15 rotate together again, and the second 2. Move the pipette to the position shown in Figure A and wait for the next new pipette to be delivered by the pipette supply device. In this case as well, the rotating member 13 and the rotating plate 15 completely close the opening 11a of the main body 11. The opening 1 formed in the lid 16 after closing
6a opens. In this way, the pipette that was also fed by the pipette feeding device is fed again. As explained above, according to the pipette feeding device of the present invention, the culture chamber in which the pipette and other devices are installed and maintained in a constant culture atmosphere is isolated from the pipette feeding device, and the pipette is not allowed to enter the room. When transferring the culture chamber, the culture atmosphere does not come into direct contact with the outside air, that is, the inside of the pipette supply device, so the atmosphere inside the culture chamber does not change when the pipette is transferred into the chamber. Since the tip does not come into contact with other objects, contamination can be completely prevented.

[Brief explanation of drawings]

FIG. 1 is a sectional view of the apparatus of the present invention, and FIG. 2 is a sectional view taken along the line 11--11. DESCRIPTION OF SYMBOLS 1... Pipette supply device, 2... Feeding device, 3... Holding arm, 4... Culture chamber, 10... Pipette, 11... Feeding device main body, 13... Rotating member, 14...
...Guide plate, 15...Rotating plate.

Claims (1)

[Claims] 1 A main body shaped like a hollow container with a bottom and having an inlet and an outlet, a guide plate having a guide groove for holding and moving a pipette fixed inside the main body, and an atmosphere at the inlet and an atmosphere at the outlet. A pipette feeding device comprising a rotating member for shielding the pipette from mixing with the atmosphere and for moving the pipette along the guide groove.