JPS5953025B2 - Centrifugal tube waste device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a waste liquid device for disposing of waste liquid in a centrifugal tube of a centrifugal separator in an automatic culture apparatus for subculturing tissues or cells.

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

However, subculturing biological tissues and cells is technically difficult, and stable culture strains have not been 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 current cell passaging methods are described below.

First, in order to carry out subculture, a predetermined number of cells are diluted with a culture solution in a culture container such as an IJ tissue, and the cells are cultured stationary in an incubator maintained at a predetermined atmosphere.

In order to check the culture state after a predetermined period of time has elapsed, a predetermined culture container is removed from the incubator and the degree of cell proliferation is examined using a microscope.

When it is confirmed that the target cells have grown to the full capacity of the container, the cells are transferred to a sterile clean bench, the culture solution in the culture container is aspirated with a pipette, and the culture solution is discarded. The remaining cells are washed by injecting buffer, and the buffer used for this washing is again aspirated and discarded.

Next, an enzyme such as trypsin is injected to release the cells that have settled and proliferated on the floor of the culture container from the culture container, and the cells are left for several minutes and then discarded.

After allowing it to stand for a few more minutes, the culture solution is injected.

This culture solution has the effect of stopping the action of trypsin.

After injecting the culture solution, stir and transfer to a centrifuge tube for centrifugation.

Aspirate and discard the centrifuged supernatant.

Next, the culture solution is injected and stirred to resuspend the cells, and the resuspended cell suspension is dispensed in fixed amounts into new culture containers.
Furthermore, a culture solution is injected to a predetermined concentration.

The culture container in which the dilution and dispensing operation has been completed in this way is taken out from the clean bench, transferred to an incubator maintained at a predetermined atmosphere, and left to stand still to proceed with the culture again.

However, the method described above has the following drawbacks.

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

For this reason, culture conditions may suddenly change when the cultured material is removed from a predetermined environmental condition such as gas atmosphere, temperature, humidity, etc., resulting in subtle changes 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 the technician's subculture operation may have a direct effect on the tissue or cells. .

In other words, this shows that it is not possible to perform standardized culture under certain conditions using conventional subculture procedures such as this.

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 when researchers conduct research on the same topic, it is often seen that researchers come to completely opposite conclusions.

Furthermore, because it takes a considerable amount of time to train technicians equipped with these cultivation techniques, there is a shortage of personnel, and researchers are unable to devote their full efforts to their original research, leading to a lack of focus on culturing techniques for branches and leaves. The reality is that you have to spend a lot of energy.

From the above, we can prevent contamination of outside air by conventional methods, eliminate the influence of human manipulation,
Further, automatic culture apparatuses are being developed for the purpose of automatically subculturing standardized tissues or cells by standardizing and unifying each operation in subculture.

The present invention is a waste liquid device for discarding supernatant liquid after centrifugation in an automatic culture device as described above, and provides a centrifugal tube waste liquid device in which the centrifuge tube is tilted and disposed of without falling. It is something to do.

The contents of the present invention will be explained in detail below based on the drawings.

In FIGS. 1 and 2, 1 is a centrifugal tube, 2 is a centrifugal tube holder, 3 is a holder presser, and 4 is rotatably supported by a support base 5 with one end of the holder presser fixed on a shaft. It is rotated by the motor 6 until the stopper pin 7 hits the stop piece 8.

The part shown at 10 is a mechanism to prevent the centrifuge tube 1 from being removed from the centrifuge tube holder 2, and 11 is a column, and 12 is a connecting rod, one end of which is attached to the column 11 by a shaft 13 so as to be rotatable. It is being

Reference numeral 14 denotes a centrifugal back press having a structure as shown in FIG. is connected to.

Reference numeral 16 is a spring attached at one end to the centrifugal backrest 14 and at the other end to a mounting shaft 17 fixed to the top of the column.

Note that 11a and 12a are stopper pieces provided on the support column 11 and the connecting rod 12, respectively.

Next, the operation of the centrifugal tube waste liquid device of the present invention will be explained with reference to the diagrams shown in FIGS. 3 to 6 which outline the movements of the connecting rod 12 and the centrifugal back presser 14.

In FIG. 1, the centrifuge tube 1 is moved by rotating the motor 6 to rotate the holder presser 3 about the shaft 4.

As a result, the tip of the centrifugal tube 1 reaches the position of the centrifugal back holder 14, and the mouth of the centrifugal tube 1 pushes against the part of the centrifugal back holder 14a.

In other words, it moves from position A to position B as shown by the chain line in FIG. .

As a result, the connecting rod 12 rotates around the shaft 13 while remaining aligned with the centrifugal back presser 14 and gradually rises in a vertical direction.

In this way, the connecting rod 12 and the centrifugal back presser 14 reach the position shown in FIG.

When the centrifugal tube 1 moves further, the centrifugal back presser 14 still moves in the same direction and the connecting rod 12 also attempts to move with it, but the connecting rod 12 hits the stopper piece 11a provided on the support column 11 and cannot move further. Since the movement of the centrifugal back presser 14 is prevented, only the centrifugal back presser 14 rotates about the shaft 15, as shown in FIG.

If the centrifuge tube 1 is further moved, it will become as shown in FIG.

In this step, the stopper pin 7 hits the stopper piece 8, and the movement of the centrifuge tube 1 is stopped.

Due to the previous movement, the tip of the centrifuge tube 1 faces diagonally downward, so the supernatant liquid in the centrifuge tube will flow down from the centrifuge tube.If a waste liquid tank is placed at an appropriate location, the supernatant liquid will be discarded into this waste liquid tank. .

Also, during this movement, the presser piece 14a of the centrifugal back presser presses the opening of the centrifuge tube 1, so that the centrifuge tube 1 does not fall out of the centrifuge tube holder 2.

After the waste liquid in the centrifuge tube is completely discarded by performing such an operation, if the centrifuge tube 1 is moved in the opposite direction, the centrifugal back presser 14 is constantly pulled by the spring 16, so that the mouth of the centrifuge tube is removed. It rotates in the opposite direction around the shaft 15 while remaining in contact with , and changes from FIG. 6 to FIG. 5 to FIG. 4, contrary to the above.

When the centrifugal back presser 14 reaches the position shown in FIG. 4, it abuts against a stopper piece 12a provided on the connecting rod 12, so that it is prevented from rotating further around the shaft 15.

At the same time, the connecting rod 12 begins to rotate around the shaft 13,
In parallel with this, the centrifugal back presser 14 also rotates, resulting in the state shown in FIG. 3.

When the centrifuge tube 1 moves further, the mouth of the centrifuge tube 1 separates from the presser piece 14a of the centrifuge back presser 14, but at this time, the centrifuge tube 1 is only slightly tilted from the vertical position, so it does not come out from the centrifuge tube holder 2. returned to the initial state.

As explained above, the liquid waste device of the present invention disposes the waste liquid in the centrifuge tube by simply tilting the centrifuge tube so that the tip faces diagonally downward while the centrifuge tube remains inserted in the holder. Moreover, even if the centrifuge tube is kept tilted to dispose of waste liquid, the centrifuge tube will not come out from the holder due to the centrifugal back holder.

[Brief explanation of drawings]

Figure 1 is a side view showing the mechanism for rotating the centrifuge tube in the liquid waste device of the present invention, Figure 2 is a front view of the device of the present invention, and Figures 3 to 6 are connecting rods that correspond to the rotation of the centrifuge tube. FIG. 7 is a perspective view of the centrifugal back press. 1... Centrifugal tube, 2... Centrifugal tube holder, 11... Strut, 12... Connecting rod, 1
4...Centrifugal back presser.

Claims (1)

[Claims] 1. In a liquid waste device in which a centrifuge tube inserted into a holder is rotated together with the holder so that the tip of the centrifuge tube is directed downward to cause the waste liquid in the centrifuge tube to fall and be disposed of, the centrifuge tube is pulled out from the holder. The presser piece has a presser piece for holding down a part of the mouth of the centrifuge tube to prevent it from coming out, and the presser piece makes almost the same movement as the mouth moves when rotating the centrifuge tube to discard waste liquid. A centrifugal tube waste liquid device characterized in that it is equipped with a centrifugal back press supported by a column.