JPH0330665A - Automatically subculturing device - Google Patents

Abstract

PURPOSE:To readily and automatically carry out subculture of microorganism, cell, etc., by opening and closing a flexible tube containing a culture medium packed therein with a number of cooks in the longitudinal direction and blending a culture medium of a microorganism with the new culture medium. CONSTITUTION:A culture medium is packed into the inside of a flexible tube 1 capable of sterilizing bacterium and made of flexible material having O2 and CO2 permeable properties and the tube is fastened at the prescribed positions with a number of and clothing cock 2 in longitudinal direction to prepare the culture tank. Then microorganism or cell is cultured in the culture tank and after definite time, culture medium is blended with new culture medium by moving a fastening position of cock in a state in which microorganism or cell is sufficiently proliferated. Thus, new culture tank is prepared to automatically carry out subculture of microorganism or cell, etc.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention is directed to microorganisms, cells, etc. applied to automatic subculturing equipment that automates preliminary culture operations to ensure cell-in (cell-1ine). This invention relates to an automatic transplantation device.

[Conventional technology]

Conventionally, all microorganisms, cells, etc. are transplanted manually using plastic or glass culture containers (bottles, flasks, jars, tubes, etc.) and pipettes. Since the transplanting operation must be performed aseptically, it is usually performed using bioclean pliers.

To date, there have been many devices that automatically transplant microorganisms, cells, etc.

[Problem to be solved by the invention]

When culturing microorganisms and cells, it is common to repeat pre-cultivation and subculture several times in order to acclimate them to the new environment.

In addition, in the case of cells that are sensitive to freezing, cultivation and subculture must be repeated just for the purpose of preserving the stock. Furthermore, transplanting operations must be carried out at a designated time, which puts a heavy burden on researchers, forcing them to work on holidays. Furthermore, since working time is limited in experiments in space, automation of experimental operations is essential.

In view of the above-mentioned state of the art, the present invention seeks to provide a device that can automatically transplant microorganisms, cells, etc.

[Means to solve the problem]

The present invention provides a microorganism characterized by comprising a flexible Y-tube filled with a culture medium, and a large number of cocks for tightening and loosening the tube, which are provided at appropriate intervals in the longitudinal direction of the tube. This is an automatic transplantation device for cells, etc.

[Effect]

Fill a tube made of flexible material with a culture medium, and tighten the tube at a predetermined position with a stopcock to create a culture tank with a fixed capacity. Microorganisms or cells are cultured in this tank, and when the microorganisms or cells have grown sufficiently, the stopcock is moved and the culture solution containing the grown microorganisms or cells and the new medium are mixed at a certain ratio. Create a new culture tank as follows. By repeating this operation, the seeds are automatically transplanted at regular intervals.

Since the device of the present invention has a simple mechanism, it is easy to automate. Further, although such operations must be performed aseptically, the apparatus of the present invention does not allow the culture solution to come into direct contact with the outside air, so there is little risk of contamination. It can also operate not only on the ground but also in zero gravity.

〔Example〕

An embodiment of the apparatus of the present invention will be described with reference to FIG.

The flexible tube 1 can be sterilized and has a 0. and Co,
A material made of a transparent material is used. Furthermore, since it is necessary to insert a syringe needle when planting or collecting the culture solution, it is preferable that the tube 1 is provided with a planting/sampling boat 3 that does not leak liquid even when the syringe needle is inserted.

Transplanting is performed by mixing the culture solution and new medium by opening and closing cocks 2 installed at appropriate intervals. At this time, the mixing ratio of the culture solution and new medium should be limited to 1:9, and if a high dilution is required, the same operation can be repeated continuously. For example, cultured culture tank 4 in FIG.
Assume that there are 10 CC of culture fluid in the cell. Moreover, the unused culture tank 5 contains a new culture medium. When trying to mix this cultured culture solution ICC and new medium 9CC,
First, close the cock 2 at the part where the cultured culture tank 4 is divided 9:1 and the cock 2 at the part where the culture medium of 9C1 is to be separated from the unused culture tank 5. Open the two cocks 2 that formed the shape.

Through these series of operations, 1 cc of cultured culture solution and 9 cc of new medium will be mixed into the new culture tank 7. An example of this state is shown in the lower diagram of FIG. The new culture tank 7 shown in the lower figure is a tank containing the cultured culture solution ICC from the previous operation and the medium 9CC from the unused culture tank 5.

Strict quantification is not required for transplanting operations! , a certain degree of quantitativeness is necessary.

In the case of the apparatus of the present invention, when attempting to dilute to 1/100, for example, it is impossible unless a very long tube is used. Therefore, a dilution of 1/100 can be achieved by performing the 1/1o dilution operation twice.

The opening/closing mechanism for the cocks 2 can be constructed by installing a large number of cocks and opening and closing them one after the other, or by using four cocks 2 in a set of two cocks 2 to open and close the tube 1 and cocks 2 while shifting their positions. There are several possible methods.

Furthermore, when culturing adherent cells, a support material such as a microcarrier may be mixed into the culture medium.

FIG. 2 shows a functional block diagram of an automatic subculturing apparatus to which the apparatus of the present invention is applied for part of the open/close cock 2 and flexible tube 1 shown in FIG. 1. This device consists of a flexible tube, an opening/closing cock, an automatic opening/closing mechanism, a control mechanism, and a stirring mechanism. Possible stirring methods include enclosing a magnetic rotor inside the flexible tube and rotating it from the outside using a Star Two, or squeezing the flexible tube from the outside. It evens out the contents and allows for proper succession. This device is also used in an incubator to control the environment (temperature, humidity, gas components) during culture.

〔Effect of the invention〕

According to the present invention, it becomes easy to automate the transplantation of microorganisms, cells, etc.

[Brief explanation of drawings]

FIG. 1 is a detailed explanatory diagram of the present invention, and FIG. 2 is a functional block diagram of an automatic subculturing apparatus to which the present invention is applied.

Claims (1)

[Claims] Automatic planting of microorganisms, cells, etc., characterized by comprising a flexible tube filled with a culture medium and a large number of cocks that can tighten or loosen the tube provided at appropriate intervals in the longitudinal direction of the tube. splicing device.