JP7269840B2 - cryopreservation container - Google Patents

Description

The present invention relates to a cryopreservation container used for long-term storage of bioresources such as iPS cells and biological samples in an ultra-low temperature range of -150°C or lower.

Public research institutes, universities, biocell banks, etc. use cryopreservation containers that use cryogenic liquefied gas such as liquefied nitrogen to freeze iPS cells, biological samples (bioresources), etc. (hereinafter simply referred to as “samples”). saved. When the rack containing the sample (sample rack) is taken out from the cryopreservation container, the rotation tray on which the sample rack is placed inside is rotated, and the target sample rack is provided on the upper wall of the cryopreservation container. It can be taken out by moving it under the opening.

In the case of a container in which the rotating operation of the rotating tray is performed manually, the rotating shaft of the rotating tray is provided with a handle for manual operation. There was a risk of frostbite and lack of oxygen when operating the handle while wearing it and visually checking the inside of the container in a cryogenic state. In addition, there is also a problem that the sample rack is hindered from being put in and taken out, such as hitting the handle when the sample rack is put in and taken out.

As a countermeasure to such a problem, Patent Document 1 discloses that "the upper end side portion of the operating shaft protrudes outside the container body through the upper wall, and the operating shaft portion that protrudes outside the container body is a motor. A cryopreservation container is disclosed that can be 'rotated' by, or artificially.
Such a cryopreservation container has a through-hole formed in the upper wall of the container body for inserting the operating shaft, and an annular seal plate for sealing between the operating shaft and the through-hole. It keeps the inside of the container airtight in a rotatable state.
According to the method of Patent Literature 1, the loading tray can be rotated outside the container main body, so the burden on the operator is reduced, and the handle for manual operation does not hinder the loading and unloading of the sample rack. do not have.

JP-A-2005-143873

However, when the rotating shaft penetrates the upper wall of the container body as in Patent Literature 1, a problem may occur in the vicinity of the above-described sealing portion.
For example, since the inside of the container is at a very low temperature, the temperature difference between the inside and the outside of the container is very large.
Such condensation travels along the rotating shaft and accumulates in the seal portion, where it is further cooled to become ice. If such ice accumulates on the seal portion, there is a problem that the rotary shaft cannot be rotated smoothly.
In addition, since the position of the opening for inserting and removing the sample rack is close to the rotating shaft, the cold air discharged from the opening when inserting and removing the sample rack and when the cryogenic liquefied gas is regularly replenished also cools the rotating shaft. This has also been a factor in dew condensation and ice build-up at the sealing portion described above. Also, cold air escaping from the opening of the container can degrade the seal.

The present invention has been made to solve this problem. An object of the present invention is to provide a cryopreservation container that can prevent accumulation.

(1) The cryopreservation container according to the present invention has an opening for putting in and taking out a sample at a position deviated from the center of the upper wall, and a lid provided in the opening so as to be openable and closable. A container main body storing gas, a rotary tray rotatably disposed in the container main body and on which a plurality of cases for storing objects to be frozen are placed, and a lower end connected to the rotary tray and an upper end has a rotating shaft that penetrates the upper wall of the container body and extends upwardly from the upper wall, and by rotating the rotating shaft, the desired cryopreserved object storage case is placed on the By rotating the rotating tray so that the position is located below the opening, it is configured to be put in and taken out from the opening, and the rotating shaft is made of a material with low thermal conductivity. and a seal mechanism for sealing the rotating shaft in a rotatable state at an upper portion of the rotating shaft, the sealing mechanism being erected on the upper wall and extending upward and into which the rotating shaft is inserted. and a bushing provided at the upper end of the rotating shaft inserting tube for hermetically and rotatably sealing the rotating shaft.

(2) Further, in the apparatus described in (1) above, the upper end portion of the rotating shaft insertion tube extends upward from the opening portion of the container body.

The cryopreservation container according to the present invention has a sealing mechanism provided with a rotating shaft insertion tube that is erected on the upper wall and extends upward, so that the rotating shaft penetrates the upper wall of the container body. Even in a cryopreservation container, the occurrence of dew condensation in the vicinity of the seal portion of the rotating shaft and the resulting ice build-up are prevented, so that the smooth rotating operation of the rotating shaft can be ensured.

It is a figure explaining the cryopreservation container concerning one embodiment of the present invention. Fig. 2 is a plan cross-sectional view of the cryopreservation container for explaining the inside of the container main body according to one embodiment of the present invention (before placement of the cryopreservation storage case). Fig. 2 is a plan cross-sectional view of the cryopreservation container for explaining the inside of the container main body according to one embodiment of the present invention (after placement of the cryopreservation storage case).

The cryopreservation container 1 according to the present embodiment, as shown in FIG. A rotary shaft 9 which is connected and whose upper end penetrates the upper wall of the container body 3 and extends above the upper wall, and a seal mechanism 11 which seals the penetrating portion while the rotary shaft 9 is rotatable. It is.
Each configuration of the cryopreservation container 1 will be specifically described below.

<Container body>
The container main body 3 is a heat insulating container with a vacuum double structure consisting of an inner tank 3a and an outer tank 3b.
A cryogenic liquefied gas 17 such as liquid nitrogen is stored in the bottom of the inner tank 3a, and the inside of the inner tank 3a is in an ultra-low temperature region of -150°C or less. The liquid level height of the cryogenic liquefied gas 17 is measured by a liquid level gauge 19 installed in the inner tank 3a, and is kept within a certain range by periodically replenishing the cryogenic liquefied gas 17. .

The upper wall of the container body 3 is provided with an opening 21 for inserting and removing the sample at a position offset from the center. The opening 21 is formed of a cylindrical neck tube or the like, and can be opened and closed by fitting and removing the cover 23 in the opening 21 .
A through-hole 25 is formed in the center of the upper wall for passing the rotating shaft 9. The through-hole 25 is a cylindrical vacuum bellows 27 provided between the inner tank 3a and the outer tank 3b. It is composed by
Further, a plurality of (four in this embodiment) casters 29 are attached to the lower portion of the outer tub 3b.

<Rotating tray>
The rotating tray 7 is for placing a cryopreserved object storage case 5 containing a cryopreserved object such as a biological sample. 2 and 3 show plan cross-sectional views of the container body 3 in the cryopreservation container 1. FIG. FIG. 2 shows the state before placing the storage case 5 for the object to be stored frozen, and FIG. 3 shows the state after placing it.
As shown in FIGS. 2 and 3, the rotary tray 7 has a circular shape in plan view, and is divided into a plurality of sections (four sections in the present embodiment) by a partition plate 31 . At the center of the rotating tray 7, a division mark 33 (a number may be used) that enables identification of each division is marked. is managed as a mark of the placement location.

The rotary tray 7 is supported by a support mechanism 35 as shown in FIG. The support mechanism 35 in this embodiment will be outlined below.
The support mechanism 35 includes a support stand 37 erected at the center of the bottom of the container body 3 , a support portion 39 fixed to the upper surface of the support stand 37 , and an annular receiver fixed to the center of the lower surface of the rotating tray 7 . A body 41, a cylindrical shaft 43 formed at the center of the rotary tray 7, and a post 45 erected at the center of the upper surface of the support portion 39 and inserted through the cylindrical shaft 43 through the receiving body 41. is.

Although detailed description is omitted, a plurality of metal spheres (not shown) are arranged between the support part 39 and the receiving body 41, and can roll in annular grooves formed in the supporting part 39 and the receiving body 41. are engaged in good condition. The support mechanism 35 supports the rotary tray 7 so as to be rotatable around the center line.

<Rotating axis>
The rotary shaft 9 has a lower end connected to the center of the rotary tray 7 (upper surface of the cylindrical shaft 43) and an upper end extending through the upper wall of the container body 3 to extend above the upper wall. , made of materials with low thermal conductivity. A through-hole 25 through which the rotating shaft 9 passes is sealed by a sealing mechanism 11, which will be described later, so that the container body 3 can be kept airtight.

The rotary shaft 9 has an operation handle 47 for manual operation on the upper portion of the outer side of the container body 3 , and by manually rotating the rotary shaft 9 using the operation handle 47 , it is connected to the lower end of the rotary shaft 9 . The rotary tray 7 can be rotated.

At this time, for example, if a mark (for example, the same mark as the division mark 33 marked on the rotary tray 7) is attached to the outside of the container body 3 of the rotary shaft 9 at a position corresponding to the division of the rotary tray 7, the lid portion 23 The compartment in which the desired cryopreserved object storage case 5 is placed can be moved to below the opening 21 without removing the , and the amount of evaporation of the cryogenic liquefied gas 17 can be suppressed, which is preferable.

<Seal mechanism>
The seal mechanism 11 includes a rotating shaft inserting tube 49 into which the rotating shaft 9 is inserted, and a bushing 51 that seals between the rotating shaft 9 and the rotating shaft inserting tube 49 while the rotating shaft 9 is rotatable. . In FIG. 1, the portion of the seal mechanism 11 is surrounded by a dashed circle and its enlarged view is shown. Each part constituting the seal mechanism 11, which is the essential part of the present invention, will be described in detail below.

≪Rotating shaft insertion tube≫
The rotating shaft insertion tube 49 is formed on the upper wall of the container main body 3, and is a metal tube that is erected in the through hole 25 and extends upward, into which the rotating shaft 9 is inserted.
The connecting portion between the lower end of the rotating shaft insertion tube 49 and the upper end of the vacuum bellows 27 is welded all around so that moisture does not enter inside.

Further, the rotating shaft insertion tube 49 has a bushing fitting 49a at its upper end. The bush mounting metal fitting 49a is a metal fitting for mounting a bush 51, which will be described later, to the rotating shaft insertion tube 49, and has an annular concave portion that matches the shape of the flange of the bush 51. As shown in FIG. As with the connecting portion with the vacuum bellows 27, the bush fitting 49a is welded all around and integrated with the rotating shaft insertion tube 49, so that external moisture does not enter from the connecting portion.

≪Bush≫
The bushing 51 is a cylindrical member having low temperature resistance and having a flange portion formed at the upper end.
The flange portion is fitted to the recess formed in the bush mounting bracket 49a and attached to the rotating shaft insertion tube 49. By inserting the rotating shaft 9 through the bush 51, the container body 3 is airtightly sealed while the rotating shaft 9 is freely rotatable. can be retained (sealed).

Further, as shown in the enlarged view of FIG. 1 , a support cover 55 that presses the outer peripheral portion of the upper surface of the flange portion of the bush 51 is fixed by a plurality of bolts 57 to the bush fitting 49 a of the rotating shaft insertion tube 49 .
The support cover 55 is a member consisting of a ring-shaped side wall and a top plate having a large circular opening. It is fixed by a plurality of bolts 57.

The opening of the top plate portion of the support cover 55 is formed to have a smaller diameter than the flange portion of the bushing 51 and a larger diameter than the rotating shaft 9 . As a result, even if the rotating shaft 9 is eccentric during the rotating operation, it does not come into contact with the support cover 55, so the bush 51 can be pressed without interfering with the rotating operation.

When the support cover 55 presses the bush 51 from above, for example, even if the pressure inside the container body 3 rises when the cryogenic liquefied gas 17 is replenished, the deformation of the bush 51 can be suppressed. Cold air leaking from between the bush 51 and the rotating shaft 9 can be reduced.
In addition, by using such a support cover 55, deterioration of the seal member (bush 51) can be suppressed compared to the conventional case where the annular seal plate is directly bolted.

The reason why dew condensation is less likely to occur on the rotary shaft 9 when the seal mechanism 11 as described above is used will be described below.
The rotating shaft inserting tube 49 is provided outside the container main body 3 and its outer peripheral surface is exposed to the atmosphere, so the gas inside the rotating shaft inserting tube 49 is warmed by the heat input from the outer peripheral surface. Along with this, the temperature of the rotating shaft 9 inside the rotating shaft insertion tube 49 also rises, so dew condensation is less likely to occur on the rotating shaft 9 outside the rotating shaft insertion tube 49 .

In addition, even if the internal pressure rises when the cryogenic liquefied gas 17 is replenished and the gas leaks from between the bush 51 and the rotating shaft 9, the temperature inside the inner tank 3a (-150 ° C.) Since the temperature rises as a result, dew condensation is less likely to occur. However, as described above, in the present embodiment, the support cover 55 is used to hold down the bush 51, so that such cold air leakage itself is less likely to occur.

Also, regarding the fact that the rotating shaft 9 was cooled by the cold air discharged from the opening 21 when the lid portion 23 was removed and the sample was put in and taken out, the dew condensation occurred. can be dealt with.
Generally, low-temperature gas moves downward, so the cool air discharged from the opening 21 also moves downward from the opening 21 . As shown in the enlarged view of FIG. 1, if the upper end of the rotary shaft insertion tube 49 is extended above the opening 21, the cold air is less likely to hit the rotary shaft 9, resulting in condensation. can be suppressed.

A procedure for taking out a sample frozen-preserved in the cryopreservation container 1 according to the present embodiment will be described below. It is assumed that liquid nitrogen is used as the cryogenic liquefied gas 17 .
First, with the lid portion 23 still attached to the opening portion 21, the operating handle 47 shown in FIG. Rotate the rotary tray 7 so that the compartment where it is located is positioned below the opening 21 . At this time, it is possible to confirm whether or not the target section is positioned below the opening 21 by a mark marked on the rotating shaft 9 .

Manual rotation tends to cause eccentricity of the rotating shaft 9, but as described above, in the present embodiment, leakage of cool air from the sealing portion due to eccentricity of the rotating shaft 9 is less likely to occur, and the leakage is very small. Even if there is, it is the nitrogen whose temperature has risen in the rotating shaft insertion tube 49, so dew condensation is unlikely to occur on the rotating shaft 9.

Next, after removing the lid portion 23 at the opening 21 of the cryopreservation container 1 and allowing it to stand in a safe place, nitrogen gas is supplied into the cryopreservation container 1 to remove the fog inside the cryopreservation container 1, Ensure visibility.
When the visibility is secured, the desired cryopreserved object storage case 5 is visually confirmed and pulled up from the opening 21 .

At this time, low-temperature nitrogen is discharged from the opening 21 from which the lid 23 has been removed. Since it is located above the opening 21, it is not cooled by the low-temperature nitrogen discharged from the opening 21, and condensation is less likely to occur.

Subsequently, the target sample is taken out from the cryopreserved object storage case 5 that has been pulled up, and the cryopreserved object storage case 5 is returned to the cryopreservation container 1 through the opening 21 and placed on the rotating tray 7 . The removed lid portion 23 is attached to the opening portion 21 to complete taking out of the sample.

As described above, according to the cryopreservation container 1 according to the present embodiment, dew condensation occurs in the vicinity of the seal portion of the rotating shaft 9 in the storage state and when the cryopreserved object storage case 5 is taken out. It is possible to prevent the accumulation of ice on the rotating shaft 9 and ensure the smooth rotation of the rotating shaft 9 .

1 Cryopreservation Container 3 Container Body 3a Inner Tank 3b Outer Tank 5 Storage Case for Cryopreserved Object 7 Rotating Tray 9 Rotating Shaft 11 Sealing Mechanism 17 Cryogenic Liquefied Gas 19 Liquid Level Gauge 21 Opening 23 Lid 25 Through Hole 27 Vacuum Bellows 29 caster 31 partition plate 33 division mark 35 support mechanism 37 support base 39 support part 41 receiver 43 cylindrical shaft 45 strut 47 operation handle 49 rotary shaft insertion tube 49a bush mounting bracket 51 bush 55 support cover 57 bolt

Claims (1)

a container body having an opening for putting in and taking out a sample at a position deviated from the center of the upper wall and a cover part provided in the opening so as to be openable and closable, and storing a cryogenic liquefied gas; a rotating tray on which a plurality of storage cases for frozen storage items are placed; and a lower end portion connected to the rotating tray and an upper end portion penetrating the upper wall of the container body to pass through the container body. and a rotary shaft extending upward from the upper wall, and by rotating the rotary shaft, the desired cryopreserved object storage case is positioned below the opening. A cryopreservation container configured to be taken in and out from the opening by rotating the rotating tray,
A sealing mechanism is provided on the upper part of the rotating shaft to seal the rotating shaft in a rotatable state,
The sealing mechanism includes a rotary shaft insertion tube that is erected on the upper wall and has an upper end that extends above the opening of the container body and into which the rotary shaft is inserted; and an upper end of the rotary shaft insertion tube. a bushing provided in the portion for airtightly and rotatably sealing the rotating shaft;
A cryopreservation container, wherein the bushing is a cylindrical member having a flange formed on its upper end, and is provided with a support cover that presses the outer circumference of the upper surface of the flange.

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