JP2024110138A - Cryopreservation Equipment - Google Patents

Abstract

To provide a cryopreservation device that is capable of suppressing attachment of frost onto a housing tool and a cryopreservation container of a sample by a simple device configuration when carrying in/out the sample from the cryopreservation container.SOLUTION: A cryopreservation device 1 comprises: a housing part 2A housing a cryopreservation container 3; a carrying in/out work space part 2B which is positioned above the housing part 2A and provided in communication with an opening 3A of the cryopreservation container 3; a dry gas supply part 4 which supplies dry gas reducing a dew point of the carrying in/out work space part 2B into the carrying in/out work space part 2B; a dry gas supply path L1 which is positioned between the dry gas supply part 4 and the carrying in/out work space part 2B; and an on-off valve 6 which is positioned in the dry gas supply path L1 and adjusts a supply amount dry gas into the carrying in/out work space part 2B, where the carrying the in/out work space part 2B is filled with dry gas to be positive pressure, and a carrying in/out work of the sample is performed between the cryopreservation container 3 and the carrying in/out work space part 2B.SELECTED DRAWING: Figure 1

Description

The present invention relates to a cryopreservation device.

Cryopreservation is commonly used as a simple method for preserving biological samples such as sperm, fertilized eggs, and cells from laboratory animals used in the development of new drugs and basic medical research. In particular, the cryopreservation method using liquid nitrogen is widely used as it is considered to be the most stable method for preserving samples for the longest period of time.

In this cryopreservation method, the biological sample is stored in an ampoule, which is then stored frozen in a cryopreservation container while still in an ampoule storage device. When the ampoule is taken in or out of the cryopreservation container, the ampoule and ampoule storage device come into contact with the air, and moisture from the air adheres to the ampoule, ampoule storage device, cryopreservation container, etc. as frost.

Therefore, in conventional cryopreservation devices (also called "glove boxes"), frost that adheres to ampoules and ampoule storage devices (hereinafter also called "ampoules, etc.") accumulates as frost and ice particles inside the cryopreservation container every time the device is loaded or unloaded. To remove this, it is necessary to periodically transfer the ampoules or other preserved samples from the cryopreservation container, then remove the liquid nitrogen from the container and dry the inside of the container, which is known as maintenance.

Patent Document 1 discloses a glove box configuration that prevents frost and ice from forming on ampoules, etc. The glove box includes a storage section that stores cryopreservation containers, and an entry/exit work space section that is connected to the top of the storage section, and the entry/exit work space section is filled with dry gas to create a positive pressure.

JP 2009-190163 A

However, in the cryopreservation device disclosed in Patent Document 1, when filling the loading/unloading work space with dry gas, it was necessary to provide an evaporator (vaporizer) that vaporizes the liquefied gas in the introduction path of the liquefied gas from the liquefied gas supply source to the loading/unloading work space.

The present invention has been proposed in light of the above-mentioned conventional circumstances, and aims to provide a cryopreservation device that, with a simple device configuration, can prevent frost from forming on the sample storage device and the cryopreservation container when samples are inserted into and removed from the cryopreservation container.

In order to solve the above problems, the present invention provides the following means.
[1] A storage unit for storing a cryopreservation container;
An entry/exit work space section located above the storage section and communicating with an opening of the cryopreservation container;
A dry gas supply unit that supplies a dry gas into the loading/unloading work space section to lower a dew point of the loading/unloading work space section;
A dry gas supply path located between the dry gas supply unit and the loading/unloading work space unit;
An on-off valve is provided in the dry gas supply path to adjust the amount of dry gas supplied to the loading/unloading work space section;
The loading/unloading work space is filled with dry gas and is under positive pressure,
A cryopreservation device that performs sample loading and unloading operations between the cryopreservation container and the loading and unloading work space section.
[2] The freezing storage device according to [1], further comprising a dew point meter for measuring a dew point within the loading/unloading work space section.
[3] The cryopreservation apparatus according to [1] or [2], further comprising a control device for controlling the opening degree of the on-off valve.
[4] The cryopreservation device according to any one of [1] to [3], further comprising a buffer tank located in the dry gas supply path and configured to temporarily store the dry gas.
[5] The cryopreservation device according to any one of [1] to [4], wherein the dry gas supply unit is a preliminary freezing device.
[6] The pre-freezing device includes a freezing storage rack for storing freezing storage boxes,
The cryopreservation device according to [5], which has a storage space for storing the cryopreservation rack and pre-freezes samples stored in the cryopreservation box.

As described above, the present invention provides a cryopreservation device that can prevent frost from forming on the sample storage device and the cryopreservation container when samples are inserted into or removed from the cryopreservation container, using a simple device configuration.

FIG. 1 is a system diagram of a cryopreservation apparatus according to one embodiment of the present invention. FIG. 2 is a front view showing the configuration of a preliminary freezing device applicable to the cryopreservation apparatus of the present embodiment. FIG. 2 is a top view showing the configuration of a preliminary freezing device applicable to the cryopreservation apparatus of the present embodiment. FIG. 2 is a front view showing the configuration of a cryopreservation rack used in the preliminary freezing device. FIG. 2 is a side view showing the configuration of a cryopreservation rack used in the preliminary freezing device. FIG. 2 is a perspective view showing the configuration of a cryopreservation box and a cryopreservation container used in the preliminary freezing device. FIG. 2 is a block diagram of a control device constituting the cryopreservation apparatus of the present embodiment. 4 is a control time chart for dry gas supply in the cryopreservation device of the present embodiment. FIG. 13 is a system diagram showing the configuration of a cryopreservation apparatus according to another embodiment of the present invention. FIG. 13 is a system diagram showing the configuration of a cryopreservation apparatus according to another embodiment of the present invention.

Hereinafter, an embodiment of the present invention will be described in detail with reference to the drawings.
In addition, the materials, dimensions, etc. exemplified in the following description are merely examples, and the present invention is not necessarily limited to them, and can be implemented with appropriate modifications within the scope that does not change the gist of the present invention.

<Freezing storage device>
First, as one embodiment of the present invention, a configuration of a cryopreservation device 1 shown in Fig. 1 will be described. Fig. 1 is a system diagram showing the configuration of the cryopreservation device 1 of this embodiment.

As shown in FIG. 1, the cryopreservation device 1 of this embodiment is generally configured to include a device main body 2, a dry gas supply unit 4, a flow control valve (on-off valve) 6, and a path (dry gas supply path) L1. It is preferable that the cryopreservation device 1 further includes a liquefied gas supply source 5, an exhaust valve 7, a dew point meter 8, a control device 9, and paths L2 to L3.

(Device body)
The device body 2 is a rectangular box with external dimensions of approximately 2000 mm in height, 1000 mm in width, and 1000 mm in depth. The device body 2 is divided vertically into two parts by a partition plate located in the center of the height direction, and is composed of a storage section 2A located at the bottom and a loading/unloading work space section (hereinafter also simply referred to as the "work space section") 2B located at the top. The size and shape of the storage section 2A and the work space section 2B are each a cube with one side of approximately 1000 mm.

The storage section 2A is configured to store a cryopreservation container 3. The partition plate that constitutes the upper surface of the storage section 2A has a circular through hole formed in its center, and the opening 3A of the cryopreservation container 3 in the storage section 2A can be seen through this through hole from the working space section 2B.

The opening 3A of the cryopreservation container 3 stored in the storage section 2A faces the working space section 2B and communicates with the working space section 2B. This allows the loading and unloading of samples, such as biological samples, to be frozen, between the cryopreservation container 3 and the working space section 2B.

The opening 3A of the cryopreservation container 3 is closed by a cap (not shown) that can be opened and closed. The cap is preferably of substantially the same shape as the opening 3A and made of a material with high thermal insulation properties.

The working space 2B is located above the storage section 2A and is provided in communication with the opening 3A of the cryopreservation container 3. Since the working space 2B is a space where the loading and unloading of samples is performed between the cryopreservation container 3 and the working space 2B, it is preferable that the working in the working space 2B is configured so that it can be seen from the outside. The working space 2B can be configured, for example, so that the front and both sides are covered with transparent acrylic resin plates. In addition, the acrylic resin plates covering the periphery of the working space 2B can be doubled to provide insulation and prevent condensation and frost on the outer surface of the acrylic resin plates.

The work space 2B is provided with an exhaust port (not shown), and the dry gas in the work space 2B is exhausted from the exhaust port as necessary. It is preferable to provide the exhaust port with a filter and a backflow prevention mechanism. This allows dry gas to flow from inside the work space 2B to outside the work space 2B, but does not allow dry gas to flow from outside the work space 2B into the work space 2B, so that the work space 2B is constantly filled with dry gas.

A dew point sensor 8a is installed in the work space 2B. This dew point sensor 8a measures the amount of moisture in the atmospheric gas in the work space 2B. A signal from the dew point sensor 8a is sent to the dew point meter 8, which calculates the dew point in the work space 2B based on the measured amount of moisture.

The configurations of the device body 2 and the cryopreservation container 3 are merely examples and are not limited thereto. Conventionally known cryopreservation devices and cryopreservation containers may be used as the device body 2 and the cryopreservation container 3. Examples include the glove box and cryopreservation container disclosed in Japanese Patent Publication No. 2009-190163 and the cryopreservation device and cryopreservation container disclosed in Japanese Patent Publication No. 6368032.

The container for storing samples when loading and unloading between the cryopreservation container 3 and the working space 2B is not particularly limited, and any conventionally known container may be used. For example, the container for storing samples may be an ampoule and an ampoule container disclosed in JP 2009-190163 A, or a vial, box, and case disclosed in JP Patent No. 6368032 A.

The dry gas supply unit 4 supplies dry gas into the working space 2B to lower the dew point of the working space 2B. Note that dry gas is a gas that does not cause frost in the environment of the working space 2B, for example, a gas with a dew point of -50°C or lower.

The dry gas supply unit 4 is connected to a liquefied gas supply source 5 via a path L3, and dry gas is derived using liquefied gas supplied from the liquefied gas supply source 5. The liquefied gas is not particularly limited as long as it is a component that has no effect when it comes into contact with the sample. Examples of liquefied gas include liquid nitrogen, liquid oxygen, and liquefied carbon dioxide. Among these, it is preferable to use liquid nitrogen from the standpoint of safety and cost.

The dry gas supply unit 4 is not particularly limited as long as it is a device or machine that uses liquefied gas supplied from the liquefied gas supply source 5 to produce dry gas. Examples of such dry gas supply units 4 include a preliminary freezing device that freezes samples before they are cryopreserved, a freeze-crushing device used in the chemical and bio-related fields, a quick-freezing device in food processing, and a cooling device such as a sub-zero treatment or cold fitting used in the heat treatment field of metal processing. Among these, a preliminary freezing device is preferably used since it is often used in close proximity to the device main body 2.

(Back-up freezing device)
Here, the configuration of the preliminary freezing device 100 applicable as the dry gas supply unit 4 constituting the cryopreservation device 1 of this embodiment will be described.
2 is a front view showing the configuration of the preliminary freezing device 100. FIG.

As shown in Figures 2 and 3, the preliminary freezing device 100 has a rectangular storage space K that stores multiple (three in this embodiment) cryopreservation racks 201, and is a programmable freezer that appropriately cools and controls the temperature of the storage space K that stores the cryopreservation racks 201 by adjusting the amount of liquid nitrogen refrigerant sprayed in accordance with a temperature control program set by a temperature regulator.

Specifically, this pre-freezing device 100 is equipped with a supply port 101 that supplies liquid nitrogen as a refrigerant to the storage space K, a spray outlet 102 that sprays the liquid nitrogen supplied through the supply port 101, a fan 103 that diffuses the liquid nitrogen sprayed from the spray outlet 102, an exhaust port 104 that exhausts the liquid nitrogen that has become a gas phase in the storage space K, a lid 105 that opens and closes the upper opening facing the storage space K, and a control unit 106 that controls each part.

In the preliminary freezing device 100, a fan 103 is disposed in the center of the rear side of the cryopreservation rack 201 placed in the storage space K. The outlet 102 is located near the fan 103 and ejects liquid nitrogen toward the fan 103. The exhaust outlet 104 is disposed to the side of the storage space K.

Here, in the preliminary freezing device 100, the supply port 101 is connected to the path L3, and liquid nitrogen, which serves as a refrigerant, is supplied from the liquefied gas supply source 5 to the accommodation space K from the supply port 101 via the path L3.
Furthermore, in the preliminary freezing device 100, the exhaust port 104 is connected to the path L1, and the liquid nitrogen that has become a gas phase in the storage space K is discharged as a dry gas from the exhaust port 104 to the path L1.

(Freezing storage rack)
Next, a specific configuration of the cryopreservation rack 201 will be described with reference to FIGS.

Note that FIG. 4 is a front view showing the configuration of the cryopreservation rack 201. FIG. 5 is a side view showing the configuration of the cryopreservation rack 201.

As shown in Figures 4 and 5, the cryopreservation rack 201 has a rack body 203 in which rack sections 202 are arranged in multiple levels (seven levels in this embodiment) in the vertical direction.

The rack body 203 is made of a metal with excellent thermal conductivity, such as aluminum, or a metal with excellent low-temperature resistance, such as stainless steel, and has multiple (seven in this embodiment) bottom plates 204 that form the bottom surface of each rack section 202, a top plate 205 that forms the top surface of the rack section located at the topmost level, and four supports 206a, 206b that form the side surfaces.

The rack body 203 has an approximately rectangular parallelepiped shape that extends vertically as a whole, by joining the multiple bottom plates 204, top plate 205, and four support columns 206a, 206b by screws, welding, etc. (in this embodiment, screws).

The four pillars 206a, 206b support the four corners of the multiple bottom plates 204 and top plates 205. Of the four pillars 206a, 206b, the two pillars 206a located on the front side of the rack body 203 are made of long plate material with a roughly I-shaped cross section and are attached to the side of the rack body 203. On the other hand, the two pillars 206b located on the rear side of the rack body 203 are made of long plate material with a roughly L-shaped cross section and are attached to the rear and side of the rack body 203.

As a result, in the cryopreservation rack 201, the cryopreservation boxes 300 stored in each of the rack sections 202 described below can be freely slid in and out in the front and rear directions from the front side of the rack body 203.

Furthermore, a pair of handles 207 are attached to both sides of the top plate 205 so as to be freely rotatable.

(Freezing storage boxes and freezing storage containers)
Next, as an embodiment of the present invention, the configuration of a cryopreservation box 300 and a cryopreservation container 400 shown in FIG. 6 will be described.
FIG. 6 is a perspective view showing the configuration of the cryopreservation box 300 and the cryopreservation container 400.

As shown in FIG. 6, the cryopreservation box 300 is made of, for example, a resin or metal box, has a rectangular shape with an open top, and has multiple storage spaces k (25 in this embodiment, 5 x 5) divided into a grid for each cryopreservation container 400 stored inside.

The cryopreservation container 400 is a cylindrical, capped container with a bottom called a vial, and after the sample is filled inside together with the culture medium, it is stored in the storage space k of the cryopreservation box 300. Note that the cryopreservation container 400 is not limited to the above-mentioned vial, and may be anything capable of storing samples.

According to the cryopreservation device 1 of this embodiment, the preliminary freezing device 100 is used as the dry gas supply unit 4, so the nitrogen gas exhausted from the preliminary freezing device 100, such as a programmable freezer, can be effectively used as dry gas. In addition, by supplying the dry gas discharged from the preliminary freezing device 100 into the working space 2B of the device body 2, it is possible to lower the dew point in the working space 2B.

As shown in FIG. 1, the path (dry gas supply path) L1 is located between the dry gas supply unit 4 and the working space 2B. One end of the path L1 is connected to the dry gas supply unit 4 (exhaust port 104 of the preliminary freezing device 100), and the other end opens into the working space 2B.

Path L1 is provided with a flow control valve 6. Path L1 also branches off into path L2 on the primary side of the flow control valve 6. Path L2 is an exhaust path for dry gas. Path L2 is provided with an exhaust valve 7, and the dry gas can be exhausted outside the system by opening the exhaust valve 7.

The flow rate control valve (on-off valve) 6 is located in the path (dry gas supply path) L1 and adjusts the amount of dry gas supplied to the working space 2B. It is preferable to use a flow rate control valve 6 that can adjust the opening from 0% to 100% continuously or stepwise in response to a control signal from the control device 9.

In the cryopreservation device 1 of this embodiment, the low-temperature liquefied gas supplied from the liquefied gas supply source 5 is vaporized in the dry gas supply section 4 to become dry gas, and the flow rate is controlled by the flow rate control valve 6 and introduced into the working space section 2B.

The dew point meter 8 measures the dew point in the working space 2B. Specifically, as shown in FIG. 1, the dew point meter 8 is capable of transmitting and receiving electrical signals to and from a dew point sensor 8a located in the working space 2B, and receives the amount of moisture in the atmospheric gas in the working space 2B measured by the dew point sensor 8a as a signal from the dew point sensor 8a. The dew point meter 8 calculates the dew point in the working space 2B based on the measured amount of moisture.

In addition, in this embodiment, it is preferable that the dew point meter 8 is capable of transmitting and receiving electrical signals to and from a dew point sensor 8b located in the storage space K of the preliminary freezing device 100 used as the dry gas supply unit 4. The amount of moisture in the atmospheric gas in the storage space K of the preliminary freezing device 100 measured by the dew point sensor 8b is received as a signal from the dew point sensor 8b. The dew point meter 8 can calculate the dew point in the storage space K based on the measured amount of moisture.

The control device 9 transmits and receives electrical signals by wire or wirelessly between the dry gas supply unit 4 (pre-freezing device 100), the flow rate control valve 6, and the dew point meter 8. The control device 9 is configured so that the dew point in the working space 2B is preset to, for example, -50°C, and when the dew point calculated by the dew point meter 8 reaches or exceeds this set dew point, the flow rate control valve 6 opens and dry gas is supplied into the working space 2B from the path L1.

The control device 9 is not particularly limited as long as it has the above-mentioned functions. The control device 9 may be configured to include a central processing unit (CPU), a memory, and a hard disk drive. The control device 9 may be provided independently (separately) from the flow control valve 6, the dew point meter 8, and the dry gas supply unit 4 (pre-freezing device 100), or may be provided as an attachment to any of these components.

<Method of using the cryopreservation device 1>
Next, a method of using (also referred to as an “operating method”) the cryopreservation apparatus 1 of this embodiment will be described using as an example a case in which the preliminary freezing apparatus 100 is used as the dry gas supply unit 4 .

First, the prefreezing device 100 is used to prefreeze a sample to be cryopreserved.
When pre-freezing a sample, first, the sample is filled into a cryopreservation container 400 together with a culture medium, and then the cryopreservation container 400 is stored in each storage space k of the cryopreservation box 300, as shown in Fig. 6. In addition, a cryopreservation box 300 is stored in each rack portion 202 of the cryopreservation rack 201, as shown in Figs. 4 and 5.

As shown in Figures 2 and 3, after the cryopreservation rack 201 is accommodated in the accommodation space K of the preliminary freezing device 100, the cooling temperature of the accommodation space K accommodating the cryopreservation rack 201 is controlled while adjusting the amount of liquid nitrogen sprayed in accordance with a temperature control program set by a temperature regulator. This makes it possible to preliminary freeze the samples filled in each cryopreservation container 400.

In the cryopreservation device 1 of this embodiment, liquid nitrogen is supplied from the liquefied gas supply source 5 to the storage space of the preliminary freezing device 100 via the path L3 and the supply port 101, and the liquid nitrogen that has become gaseous in the storage space K is discharged as dry gas from the exhaust port 104 to the path L1.

Next, the sample pre-frozen by the pre-freezing device 100 is frozen and stored in the device body 2 .
When the pre-frozen sample is cryopreserved, as shown in Fig. 1, the sample is placed inside the cryopreservation container 3 through the opening 3A of the cryopreservation container 3 from the working space 2B of the device body 2, and then the opening 3A of the cryopreservation container 3 is closed with a cap (not shown). The inside (inner tank) of the cryopreservation container 3 is filled with an appropriate amount of low-temperature liquefied gas such as liquefied nitrogen.

Next, a dry gas such as nitrogen gas is introduced into the working space 2B of the device body 2 to set the pressure inside the working space 2B above atmospheric pressure or above the outside air pressure, thereby preventing outside air from entering the device body 2. For example, the pressure inside the working space 2B may be set to a positive pressure that is about 0.1 to 10% higher than the outside air pressure. However, if it is possible to prevent outside air from entering the device body 2, the pressure inside the working space 2B does not necessarily have to be a positive pressure, and may be set to the same as the outside air pressure.

There are two methods for supplying dry gas into the working space 2B: one is to supply nitrogen gas from the cryopreservation container 3 into the working space 2B, and the other is to supply dry gas into the working space 2B from the path L1 by opening the flow rate control valve 6. Either method keeps the working space 2B constantly filled with dry gas.
Hereinafter, a method for supplying the dry gas from the path L1 into the working space portion 2B with the flow rate control valve 6 in an open state will be specifically described.

(Aspect 1)
FIG. 7 is a block diagram of the control device 9 constituting the cryopreservation device 1 of this embodiment.
As one aspect of the method of supplying dry gas into the working space 2B, in the cryopreservation apparatus 1 of this embodiment, as shown in Figures 1 and 7, the moisture amount in the atmospheric gas in the working space 2B measured by a dew point sensor 8a located in the working space 2B of the apparatus main body 2 is received by a dew point meter 8, and the dew point meter 8 calculates the dew point in the working space 2B based on the measured moisture amount. The dew point meter 8 transmits the calculated dew point in the working space 2B to the control device 9 as an electric signal.
The control device 9 presets the dew point in the work space 2B to, for example, -50°C, and when the dew point calculated by the dew point meter 8 becomes equal to or higher than this set dew point, it sends a control signal to the flow control valve 6.
The flow rate control valve 6 adjusts the opening degree in accordance with a control signal sent from the control device 9. As a result, the dry gas discharged from the preliminary freezing device 100 is supplied into the working space portion 2B through the path L1.

1 and 7, in the cryopreservation apparatus 1 of this embodiment, the moisture amount in the atmospheric gas in the storage space K measured by the dew point sensor 8b located in the storage space K of the preliminary freezing device 100 is received by the dew point meter 8, and the dew point meter 8 calculates the dew point in the storage space K based on the measured moisture amount. The dew point meter 8 transmits the calculated dew point in the storage space K to the control device 9 as an electrical signal.
The control device 9 presets the dew point within the storage space K to, for example, −50° C., and when the dew point calculated by the dew point meter 8 becomes equal to or higher than this set dew point, it sends a control signal to the flow rate regulating valve 6 .
The flow rate control valve 6 adjusts the opening degree in accordance with a control signal sent from the control device 9. As a result, the dry gas discharged from the preliminary freezing device 100 is supplied into the working space portion 2B through the path L1.

In addition, in the freezing preservation device 1 of this embodiment, as shown in Figures 2, 3 and 7, an opening/closing sensor 4b is provided on the lid body 105 of the preliminary freezing device 100, and when the lid body 105 is opened and the storage space K is released, the opening/closing sensor 4b transmits an electrical signal to the control device 9.
When the control device 9 receives an electrical signal from the open/close sensor 4b notifying that the accommodation space K is in an open state, it transmits a control signal to the flow rate adjustment valve 6 to close the path L1.
The flow rate control valve 6 adjusts the opening degree to be fully closed in accordance with a control signal transmitted from the control device 9. This stops the supply of dry gas from the path L1 into the working space portion 2B.

In addition, in this embodiment 1, a thermometer 4a may be used instead of the dew point sensor 8b located in the accommodation space K of the preliminary freezing device 100.
In this case, the temperature inside the storage space K measured by the thermometer 4 a located inside the storage space K of the preliminary freezing device 100 is transmitted to the control device 9 as an electrical signal.
The control device 9 presets the temperature inside the accommodation space K to, for example, −20° C., and transmits a control signal to the flow rate adjustment valve 6 when the temperature measured by the thermometer 4 a falls below this set temperature.
The flow rate control valve 6 adjusts the opening degree in accordance with a control signal sent from the control device 9. As a result, the dry gas discharged from the preliminary freezing device 100 is supplied into the working space portion 2B through the path L1.

(Aspect 2)
FIG. 8 is an example of a time chart for controlling the supply of dry gas to the cryopreservation apparatus 1 of this embodiment.
First, as shown in Figures 1 and 8, the operation of the preliminary freezing device 100 is started, and liquid nitrogen is started to be supplied from the liquefied gas supply source 5 to the preliminary freezing device 100 to start cooling the storage space K (time 0 in Figure 8). At the same time, gas is started to be discharged from the storage space K to the path L1. At this time, the control device 9 closes the flow rate control valve 6 of the path L1 (opening degree 0) and opens the exhaust valve 7 of the path L2. As a result, the gas discharged from the storage space K to the path L1 is exhausted from the exhaust valve 7 to the outside of the system.

Next, when 20 minutes have passed since the start of cooling and it has been confirmed that the temperature of the storage space K has reached -20°C, the control device 9 closes the exhaust valve 7 of the path L2 (opening degree 0) and controls the flow rate control valve 6 of the path L1 to a predetermined opening degree. As a result, the gas led from the storage space K to the path L1 is supplied as dry gas into the working space 2B of the device body 2.

Next, 100 minutes after the start of cooling, the temperature of the storage space K reaches -80°C. The amount of liquid nitrogen supplied from the liquefied gas supply source 5 to the preliminary freezing device 100 is controlled to maintain this temperature. As a result, the amount of gas supplied from the storage space K to the path L1 is reduced.

Then, when 120 minutes have elapsed since the start of cooling, the supply of liquid nitrogen from the liquefied gas supply source 5 to the pre-freezing device 100 is stopped, and the operation of the pre-freezing device 100 is terminated. Next, the lid 105 opens to open the storage space K, and the pre-frozen sample is removed from the storage space K. At this time, the control device 9, which has received a signal to open the lid 105, closes the flow control valve 6 of path L1 (opening degree 0) and opens the exhaust valve 7 of path L2. As a result, the gas led from the storage space K to path L1 is exhausted from the exhaust valve 7 to the outside of the system.

Thus, in this embodiment 2, when the internal temperature of the pre-freezing device 100 (the temperature of the storage space K) reaches a predetermined temperature, the flow control valve 6 of path L1 is opened to supply dry gas into the working space 2B. When the internal temperature reaches the predetermined temperature, air is discharged from the interior (storage space K) and the dew point inside the interior drops. At this time, the flow control valve 6 of path L1 is opened to supply dry gas into the working space 2B. The predetermined temperature can be set to, for example, -20°C.

In this embodiment 2, the flow control valve 6 of the path L1 is operated based on the internal temperature of the prefreezing device 100, but the operation may be based on the dew point temperature instead of the internal temperature.

In the cryopreservation device 1 of this embodiment, the procedure for removing a sample from the device body 2 is the reverse of the procedure for cryopreserving a sample in the device body 2 described above. That is, as shown in FIG. 1, a cap (not shown) closing the opening 3A of the cryopreservation container 3 is removed, and the sample stored inside the cryopreservation container 3 is removed from the opening 3A into the working space 2B, and then removed to the outside of the device body 2.

Here, when loading and unloading samples between the cryopreservation container 3 and the working space 2B, a conventionally known method may be applied. For example, as disclosed in JP 2009-190163 A, a worker may insert his/her hands into gloves in the working space 2B and load and unload samples between the cryopreservation container 3 and the working space 2B. Also, as disclosed in JP 2009-190163 A and Japanese Patent No. 6368032 A, an automatic transport device may be used to automatically load and unload samples between the cryopreservation container 3 and the working space 2B.

As described above, according to the cryopreservation device 1 of this embodiment, by using a preliminary freezing device 100 such as a programmable freezer as the dry gas supply unit 4, the nitrogen gas exhausted from the preliminary freezing device 100 can be supplied as dry gas into the working space 2B of the device body 2. This eliminates the need to provide an evaporator or the like in the liquefied gas supply path connected to the liquefied gas supply source, simplifying the configuration of the entire device.

In addition, according to the cryopreservation device 1 of this embodiment, the nitrogen gas exhausted from the preliminary freezing device 100 can be supplied as dry gas into the working space 2B of the device body 2, thereby lowering the dew point in the working space 2B. Therefore, the nitrogen gas exhausted from the preliminary freezing device 100 can be effectively utilized, and frost formation on the sample storage device and the cryopreservation container 3 can be suppressed.

In addition, according to the cryopreservation device 1 of this embodiment, by providing a control device 9, dry gas can be automatically supplied before the dew point in the working space 2B of the device body 2 becomes high, so that a dry state in the working space 2B can be easily maintained.

In addition, according to the cryopreservation device 1 of this embodiment, when the preliminary freezing device 100 is used as the dry gas supply unit 4, the gas in the preliminary freezing device 100 is confirmed to be dry before being supplied as dry gas to the working space section 2B, thereby preventing a gas with a high dew point from being supplied to the working space section 2B.

In addition, according to the cryopreservation device 1 of this embodiment, when the preliminary freezing device 100 is used as the dry gas supply unit 4, the open/close state of the flow control valve 6 located on the path L1 can be controlled according to the open/close state of the lid body 105 of the preliminary freezing device 100, so that it is possible to prevent a gas with a high dew point from being supplied into the working space portion 2B.

The present invention is not necessarily limited to the above-described embodiment, and various modifications can be made without departing from the spirit of the present invention.
For example, in the cryopreservation device to which the present invention is applied, the samples stored inside the cryopreservation container 3 are not necessarily limited to those that have been pre-frozen, but may be samples in other states.

The cryopreservation device to which the present invention is applied may also be configured to include a buffer tank for temporarily storing the dry gas.

FIG. 9 is a system diagram showing the configuration of a cryopreservation device 21 according to another embodiment of the present invention.
As shown in Fig. 9, the cryopreservation apparatus 21 differs in configuration from the above-described cryopreservation apparatus 1 in that a path (dry gas supply path) L1 is provided with a buffer tank 20 and a flow control valve 22. Note that the cryopreservation apparatus 21 shown in Fig. 9 does not include the dew point meter 8 and the control device 9. Therefore, the same components as those in the cryopreservation apparatus 1 are denoted by the same reference numerals, and their description will be omitted.

The buffer tank 20 is located in the path L1 and is a container that temporarily stores the dry gas flowing through the path L1. By providing the buffer tank 20 in the path L1, the amount of dry gas supplied to the secondary side can be stabilized even if the flow rate of the dry gas discharged from the dry gas supply unit 4 to the path L1 is not constant.

The flow rate control valve 22 is located in path L1 on the primary side of the buffer tank 20. The flow rate control valve 22 may have the same function as the flow rate control valve 6. By providing the flow rate control valve 22 in path L1, the dry gas can be reliably stored in the buffer tank 20.

As described above, the cryopreservation device 21 is configured to include a buffer tank 20 and a flow rate control valve 22 in the path L1, so that dry gas can be stably supplied to the working space 2B of the device body 2.

In addition, the number of dry gas supply units 4 and device bodies 2 in the cryopreservation device to which the present invention is applied is not limited. The cryopreservation device to which the present invention is applied may be configured as follows: (1) having two or more dry gas supply units and one device body; (2) having one dry gas supply unit and two or more device bodies; or (3) having two or more dry gas supply units and two or more device bodies.

FIG. 10 is a system diagram showing the configuration of a cryopreservation device 31 according to another embodiment of the present invention.
As shown in Fig. 10, the cryopreservation apparatus 31 is different in configuration from the cryopreservation apparatus 21 shown in Fig. 9 in that it includes two or more dry gas supply units 4 (preliminary freezing apparatuses 100) and two or more apparatus bodies 2. Therefore, components common to the cryopreservation apparatus 1 and the cryopreservation apparatus 21 are given the same reference numerals and descriptions thereof will be omitted.

As described above, the freezing preservation device 31 is equipped with two or more dry gas supply units 4 (preliminary freezing devices 100), and dry gas is discharged from one of the dry gas supply units 4 to path L1, thereby ensuring the supply of dry gas.
In addition, since the freezing preservation device 31 is equipped with a buffer tank 20, dry gas can be stably supplied into the working space 2B of the secondary side device body 2 even if the flow rate of the dry gas is not constant.
Furthermore, according to the cryopreservation device 31, since it has two or more device bodies 2, the dry gas stored in the buffer tank 20 can be effectively utilized.

Furthermore, the method of use (operation) of the cryopreservation device to which the present invention is applied is not particularly limited. In the above-described embodiment, when using the cryopreservation device 1, mode 1 and mode 2 are exemplified as the method of supplying dry gas from path L1 into the working space portion 2B with the flow control valve 6 in an open state, but these may be performed separately or simultaneously.

1, 21, 31 Cryopreservation device 2 Device body 2A Storage section 2B Storage and retrieval work space section (work space section)
3 Cryopreservation container 3A Opening 4 Dry gas supply unit 4a Thermometer 4b Opening/closing sensor 5 Liquefied gas supply source 6 Flow rate control valve (opening/closing valve)
7 Exhaust valve 8 Dew point meter 8a, 8b Dew point sensor 9 Control device 20 Buffer tank 22 Flow rate adjustment valve 100 Pre-freezing device 101 Supply port 104 Exhaust port 201 Cryopreservation rack 300 Cryopreservation box 400 Cryopreservation container K Storage space L1 Path (dry gas supply path)
L2-L3 route

Claims (6)

A storage unit for storing a cryopreservation container;
An entry/exit work space section located above the storage section and communicating with an opening of the cryopreservation container;
A dry gas supply unit that supplies a dry gas into the loading/unloading work space section to lower a dew point of the loading/unloading work space section;
A dry gas supply path located between the dry gas supply unit and the loading/unloading work space unit;
An on-off valve is provided in the dry gas supply path to adjust the amount of dry gas supplied to the loading/unloading work space section;
The loading/unloading work space is filled with dry gas and is under positive pressure,
A cryopreservation device that performs sample loading and unloading operations between the cryopreservation container and the loading and unloading work space section. The cryopreservation device according to claim 1, further comprising a dew point meter for measuring the dew point within the loading/unloading work space. The cryopreservation device according to claim 1, further comprising a control device that controls the opening degree of the on-off valve. The cryopreservation device according to claim 1, further comprising a buffer tank located in the dry gas supply path and configured to temporarily store the dry gas. The cryopreservation device according to any one of claims 1 to 4, wherein the dry gas supply unit is a preliminary freezing device. The pre-freezing device includes a freezing storage rack for storing freezing storage boxes,
6. The cryopreservation device according to claim 5, further comprising a storage space for storing the cryopreservation rack, and for pre-freezing the samples stored in the cryopreservation box.

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