A vitrification freezing tool and method of using the same

Abstract

The present invention discloses a vitrification freezing tool, which includes a connector and a carrier rod, and also discloses a method for using the vitrification freezing tool, which includes the following steps: S1, inserting biological materials; S2, adding treatment liquid; S3, discharging treatment liquid; S4, vitrification freezing and installing sleeve; S5, cryopreservation. The beneficial effects achieved by the present invention: the pretreatment and cryopreservation are integrated, the operation steps are simplified, and the biological materials are convenient to process. At the same time, the integration of pretreatment and cryopreservation is conducive to the batch and automated management of biological materials. The probability of damage and contamination of biological materials during transportation is reduced, and the biological materials on the carrier rod can be protected to avoid contamination during the batch processing of the equipment, thereby ensuring the reliability of the experimental process.

Description

Technical Field

[0001] The present invention relates to the technical field of freezing tools, and in particular to a vitrification freezing tool and a method for using the same. Background Art

[0002] Vitrification is a cryopreservation technique for biological materials. Vitrification uses liquid nitrogen to rapidly cool biological materials and inhibits the formation of ice crystals. After vitrification, biological materials are stored in an ultra-low temperature environment.

[0003] The Chinese utility model patent with the announcement number CN214339670U discloses a biological material processing device. The utility model includes a shell, and the upper end of the shell is provided with a liquid exchange chamber, a liquid discharge chamber and a plurality of liquid guide channels, and the two ends of each liquid guide channel are respectively connected to the liquid exchange chamber and the liquid discharge chamber to realize the liquid exchange between the liquid exchange chamber and the liquid discharge chamber; the lower end of the liquid exchange chamber is connected to a discharge hole, and the discharge hole is connected to the lower end of the shell. After the biological material is processed in the liquid exchange chamber, it is discharged downward to the carrier rod through the discharge hole, and the carrier rod and the embryos thereon can be transferred for subsequent vitrification freezing operation.

[0004] The above-mentioned biological material processing device still has the following problems during use: 1. After the biological material is processed in the liquid exchange chamber, the biological material is transferred to the carrier rod, and then the carrier rod carrying the biological material is transported to the freezing equipment for cryopreservation. However, it is not convenient for batch and automated processing to store the biological material on the surface of the carrier rod. During the transfer process to the carrier rod, the operator needs to check and confirm that the biological material has been discharged onto the carrier rod, which makes it difficult to achieve batch and automated pretreatment and freezing; 2. When the carrier rod and the biological material thereon are vitrified and frozen by liquid nitrogen, the biological material is easily contaminated; 3. The biological material needs to pass through the liquid exchange chamber, the discharge hole and the carrier rod in turn. When the biological material is repeatedly transferred, it is easy to make an error operation, which increases the probability of damaging the biological material; 4. In addition, the operation steps are cumbersome, which causes inconvenience to the processing of the biological material. Summary of the invention

[0005] In order to solve the above technical problems, the purpose of the present invention is to provide a vitrification freezing tool, which includes a connector and a carrier rod, and also provides a method for using the vitrification freezing tool, the method including the following steps: S1, inserting biological material; S2, adding treatment liquid; S3, discharging treatment liquid; S4, vitrification freezing and installing sleeve; S5, cryopreservation. The vitrification freezing tool has the advantages of facilitating the automation of pretreatment and freezing, preventing biological material from being contaminated, reducing the probability of damaging biological material, and being easy to operate.

[0006] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention is as follows:

[0007] A vitrification freezing tool comprises a connector and a carrier rod, wherein the carrier rod is fixedly mounted on the connector, the carrier rod is provided with a processing tank and a liquid adding tank, the carrier rod is provided with a liquid conducting channel, the two ends of the liquid conducting channel are respectively connected to the processing tank and the liquid adding tank, the carrier rod is provided with a liquid discharge port, the liquid discharge port is communicated with the processing tank, the connector is detachably mounted with a sleeve, the connector is provided with a through-port, and the connector is fixedly mounted with a filter assembly that blocks the through-port.

[0008] Through such a setting: the integration of pretreatment and cryopreservation is achieved, the operation steps are simplified, and the processing of biological materials is convenient. At the same time, the integration of pretreatment and cryopreservation is conducive to the batch and automated management of biological materials. It can greatly reduce the probability of damage and contamination of biological materials during transportation, and play a role in protecting biological materials. The setting of the sleeve can protect the biological materials on the carrier rod, avoid contamination during the batch processing of the equipment, and ensure the reliability of the experimental process. During liquid nitrogen processing and storage, the filter component can perform bidirectional filtration on liquid nitrogen.

[0009] Preferably, the filter assembly adopts one or more of stainless steel filter membrane, polymer filter membrane and ceramic membrane, and the material of the sleeve and the carrier rod adopts one or more of polyethylene, polypropylene, polystyrene, cycloolefin copolymer and acrylic.

[0010] Through such an arrangement, the function of filtering liquid nitrogen through the filter component is achieved.

[0011] Preferably, it further comprises a connecting portion, which is arranged on the connecting body.

[0012] Such an arrangement makes it easy to carry the vitrification freezing tool through the connection part, facilitates connection and movement, and is conducive to batch processing.

[0013] Preferably, the connecting portion is provided with a connecting groove, and an extension rod cooperating therewith is installed in the connecting groove.

[0014] Through such an arrangement, the entire vitrification freezing tool can be driven to move through the connection part, and the extension rod can be arranged to facilitate the picking up and operation.

[0015] Preferably, the inner contour of the cross section of the connecting groove is any one of "star-shaped", "hexagonal", "rectangular", "triangular" and "elliptical".

[0016] Through such an arrangement, the device can be suitable for different extension rods, thereby improving applicability.

[0017] Preferably, the carrier rod is provided with a drainage port, which is connected to the processing tank, the position height of the liquid guiding channel is higher than the drainage port, the width of the drainage port is set to a size that can hinder the passage of biological materials, and the setting of the drainage port width includes one or a combination of preventing the passage of biological materials by forming a restriction by setting the width size or preventing the passage of biological materials by forming a restriction by liquid surface tension.

[0018] Through such an arrangement, the drain port automatically absorbs excess liquid through capillary phenomenon, thereby reducing the amount of droplets, facilitating vitrification of biological materials, and effectively restricting biological materials, preventing them from passing through the drain port, so that the biological materials are preserved in the processing tank.

[0019] Preferably, the processing tank is provided with a first inclined surface and a second inclined surface, and the lowest positions of the first inclined surface and the second inclined surface are both located at the drain port.

[0020] With such an arrangement, the treatment liquid and biological material can be concentrated at the liquid discharge port by gravity, which facilitates the contact between the biological material and the treatment liquid, and facilitates the discharge of the treatment liquid from the treatment tank through the liquid discharge port.

[0021] Preferably, the liquid discharge port is located on a side of the treatment tank close to the liquid addition tank.

[0022] Through such an arrangement, it is convenient for the liquid in the liquid adding tank to flow to the biological material, and it is convenient for the biological material to contact with the treatment liquid, so as to facilitate the treatment of the biological material.

[0023] A method for using a vitrification freezing tool, using a vitrification freezing tool, the vitrification freezing tool comprising a connector and a carrier rod, the carrier rod being fixedly mounted on the connector, the carrier rod being provided with a processing tank and a liquid adding tank, the carrier rod being provided with a liquid conducting channel, the two ends of the liquid conducting channel being respectively connected to the processing tank and the liquid adding tank, the carrier rod being provided with a liquid discharge port, the liquid discharge port being communicated with the processing tank, the connector being detachably mounted with a sleeve, the connector being provided with a through-port, and the connector being fixedly mounted with a filter assembly blocking the through-port;

[0024] The method comprises the following steps:

[0025] S1. Place the biomaterial: remove the sleeve from the connector and place the biomaterial in the treatment tank;

[0026] S2, adding treatment liquid: adding the treatment liquid of the biological material into the liquid adding tank, wherein the treatment liquid enters the treatment tank through the liquid guiding channel and contacts the biological material;

[0027] S3, discharging the treatment liquid: after the biological material is treated, the treatment liquid in the treatment tank is discharged from the treatment tank through the addition tank, so that the biological material remains in the treatment tank;

[0028] S4, vitrification and sleeve installation: liquid nitrogen is placed in the sleeve, a carrier rod containing biological materials is inserted into the liquid nitrogen in the sleeve, and the sleeve is installed on the connector;

[0029] S5. Cryopreservation: Liquid nitrogen is allowed to enter the casing through the through-hole, and the liquid nitrogen is filtered through the filter assembly to avoid mutual contamination between biological materials. The liquid nitrogen in the casing is discharged through the through-hole when it is gasified.

[0030] Through such a setting: the integration of pretreatment and cryopreservation is achieved, the operation steps are simplified, and the processing of biological materials is convenient. At the same time, the integration of pretreatment and cryopreservation is conducive to the batch and automated management of biological materials. It can greatly reduce the probability of damage and contamination of biological materials during transportation, and play a role in protecting biological materials. The setting of the sleeve can protect the biological materials on the carrier rod, avoid contamination during the batch processing of the equipment, and ensure the reliability of the experimental process. During liquid nitrogen processing and storage, the filter component can perform bidirectional filtration on liquid nitrogen.

[0031] Preferably, the step S1 further includes the following steps:

[0032] A plurality of vitrification tools are prepared, sleeves on the plurality of vitrification tools are removed from the connector, and biological materials are placed in the plurality of processing tanks respectively.

[0033] After step S1 and before step S2, the following steps are also included:

[0034] S1.1. Batch processing: Repeat step S1 multiple times to obtain multiple vitrification freezing tools containing biological materials.

[0035] Through such a setting: biological materials can be added in batches to multiple vitrification freezing tools, and the biological materials in multiple vitrification freezing tools can be processed in batches, which has the advantage of high processing efficiency and is conducive to batch and automated management of biological materials.

[0036] Compared with the prior art, the present invention has achieved beneficial technical effects:

[0037] 1. After the treatment tank completes the treatment of the biological material, the excess treatment liquid is discharged through the discharge port by capillary action to achieve the minimum droplet amount. The sleeve is installed on the connector and the sleeve covers the carrier rod, and then the vitrification freezing tool is connected to the carrier rod for cryopreservation. After the biological material is processed in the treatment tank, there is no need to take the biological material out of the treatment tank. The vitrification freezing tool and the biological material inside are directly frozen to achieve the integration of pretreatment and cryopreservation, simplify the operation steps, and facilitate the treatment of biological materials. At the same time, the integration of pretreatment and cryopreservation is conducive to the realization of batch and automated management of biological materials.

[0038] 2. From the use of treatment liquid to treat the biological material to the freezing and thawing of the biological material, there is no need to remove the biological material from the treatment tank, which can greatly reduce the probability of damage and contamination of the biological material during transportation and protect the biological material. After the sleeve is installed on the connector, multiple vitrification tools can be transported at the same time through the transport equipment, which can realize batch cryopreservation of biological materials.

[0039] 3. The setting of the sleeve can protect the biological materials on the carrier rod and prevent them from being contaminated during the batch processing of the equipment, thus ensuring the reliability of the experimental process. During liquid nitrogen processing and storage, the filter component can filter the liquid nitrogen in both directions. When the liquid nitrogen enters the inside of the sleeve through the through-hole, the filter component filters the liquid nitrogen, which can prevent the contaminants in the liquid nitrogen from entering the sleeve and causing contamination of the biological materials, thereby preventing the biological materials from being contaminated. The liquid nitrogen can also be filtered through the filter component to prevent the liquid nitrogen outside the sleeve from being contaminated, and to prevent cross-contamination of biological materials in different vitrification freezing tools. And when the sample takes out the liquid nitrogen and moves in the atmospheric environment, the vaporized liquid nitrogen can be discharged from the through-hole to ensure safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 is a schematic structural diagram of a vitrification freezing tool in Example 1 of the present invention;

[0041] Figure 2 is a schematic structural diagram of a carrier rod in Embodiment 1 of the present invention;

[0042] Figure 3 is a schematic structural diagram of the filter assembly in Example 1 of the present invention;

[0043] Figure 4 is a schematic structural diagram of a connecting portion in Embodiment 1 of the present invention;

[0044] Figure 5 It is a schematic flow chart of a method for using a vitrification freezing tool in Example 1 of the present invention;

[0045] Figure 6 is a schematic structural diagram of a vitrification freezing tool in Example 2 of the present invention;

[0046] Figure 7 is a schematic structural diagram of a vitrification freezing tool in Example 3 of the present invention;

[0047] Figure 8 is a schematic structural diagram of a vitrification freezing tool in Example 4 of the present invention;

[0048] Fig. 9 is a schematic structural diagram of a vitrification freezing tool in Example 5 of the present invention;

[0049] Fig.10 is a schematic structural diagram of a vitrification freezing tool in Example 6 of the present invention;

[0050] Fig.11 is a schematic structural diagram of a vitrification freezing tool in Example 7 of the present invention;

[0051] Fig.12 It is a schematic structural diagram of a vitrification freezing tool in Example 8 of the present invention.

[0052] The technical features indicated by the reference numerals are as follows:

[0053] 11. Connector; 12. Carrier rod; 13. Processing tank; 14. Liquid adding tank; 15. Liquid guiding channel; 16. Liquid discharge port; 17. First inclined surface; 18. Second inclined surface; 21. Sleeve; 22. Through port; 23. Filter assembly; 24. First filter element; 25. Second filter element; 31. Connecting portion; 32. Connecting tank; 41. Extension rod. DETAILED DESCRIPTION

[0054] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments, but the scope of protection claimed in the present invention is not limited to the following specific embodiments.

[0055] Embodiment 1:

[0056] refer to Figures 1 to 4A vitrification freezing tool comprises a connector 11, a carrier rod 12 and a connecting portion 31, wherein the carrier rod 12 is fixedly mounted on the connector 11, the carrier rod 12 is provided with a processing tank 13 and a liquid adding tank 14, the carrier rod 12 is provided with a liquid conducting channel 15, both ends of the liquid conducting channel 15 are respectively connected to the processing tank 13 and the liquid adding tank 14, the carrier rod 12 is provided with a drainage port 16, the drainage port 16 is communicated with the processing tank 13, the drainage port 16 runs through the carrier rod 12, the width of the drainage port 16 is set to a size that can hinder the passage of biological materials and allow the processing liquid to pass through, and the setting of the width of the drainage port 16 includes preventing the passage of biological materials by setting a width size to form a restriction and preventing the passage of biological materials by forming a restriction by liquid surface tension. The processing tank 13 is provided with a first inclined surface 17 and a second inclined surface 18. The first inclined surface 17 and the second inclined surface 18 are inclined perpendicularly on the carrier rod 12. The lower end of the first inclined surface 17 is inclined in the length direction of the carrier rod 12, and the lower end of the second inclined surface 18 is inclined in the width direction of the carrier rod 12. The lowest positions of the first inclined surface 17 and the second inclined surface 18 are both located at the drain port 16.

[0057] The sleeve 21 is made of a low temperature resistant material. The support rod 12 is made of a high heat transfer, high transparency, and low temperature resistant material, which is convenient for microscopic observation of biological materials on the processing tank 13, the first inclined surface 17, and the second inclined surface 18. The sleeve 21 and the support rod 12 can be made of any one of polyolefins such as polyethylene, polypropylene, polyphenylene series such as polystyrene, cycloolefin copolymer, and acrylic.

[0058] The drain port 16 is located on one side of the treatment tank 13 close to the liquid addition tank 14. The liquid guiding channel 15 is located in the middle of the second inclined surface 18, and the drain port 16 is located at the bottom of the second inclined surface 18, so that the height of the liquid guiding channel 15 is slightly higher than the drain port 16, so as to avoid the liquid in the treatment tank 13 from being completely drained, thereby preventing the cells in the biological material from being damaged and protecting the biological material.

[0059] The connecting part 31 is arranged on the connecting body 11, and the connecting part 31 is fixedly connected to the connecting body 11. The connecting part 31 is provided with a connecting groove 32, and the inner contour of the cross-section of the connecting groove 32 is an "octagonal star" as a whole. The connecting groove 32 is installed with an extension rod 41 that matches it. The vitrification freezing tool is driven to move by the extension rod 41, which can achieve the effect of convenient operation.

[0060] The connector 11 is detachably mounted with a sleeve 21, and the connector 11 is provided with a plurality of through-holes 22. The connector 11 is fixedly mounted with a filter assembly 23 that blocks the through-holes 22. The filter assembly 23 includes a first filter element 24 and a second filter element 25, and the first filter element 24 and the second filter element 25 are respectively arranged at both ends of the through-hole 22. The filter assembly 23 uses one or more of a stainless steel filter membrane, a polymer filter membrane, and a ceramic membrane. In this embodiment, the filter assembly 23 uses a stainless steel filter membrane to achieve the function of filtering liquid nitrogen through the filter assembly 23.

[0061] refer to Figure 5 A method for using a vitrification freezing tool, using a vitrification freezing tool, the method comprises the following steps:

[0062] S1. Place biological material: prepare a vitrification tool without biological material, remove the sleeve 21 from the connector 11 , and place the biological material in the processing tank 13 .

[0063] S1.1. Batch processing: Repeat step S1 multiple times to obtain multiple vitrification freezing tools containing biological materials.

[0064] S2. Adding treatment liquid: Use a spray gun to add the treatment liquid of the biological material into the liquid adding tanks 14 of different vitrification freezing tools at the same time. The treatment liquid enters the treatment tank 13 through the liquid guide channel 15 and contacts with the biological material.

[0065] S3, draining the treatment liquid: After the biological material is processed, the treatment liquid in the treatment tank 13 is discharged from the treatment tank 13 through the liquid addition tank 14, and the discharge port 16 discharges the excess liquid through the capillary phenomenon, so that the biological material is retained in the treatment tank 13, effectively reducing the excess liquid in the treatment tank 13, and ensuring that the biological material can be quickly cooled and frozen.

[0066] S4, vitrification and sleeve installation: Liquid nitrogen is placed in the sleeve 21 , the carrier rod ( 12 ) containing the biological material is quickly inserted into the liquid nitrogen in the sleeve 21 , and the sleeve 21 is installed on the connector 11 .

[0067] S5. Cryopreservation: Allow liquid nitrogen to enter the sleeve 21 through the through-hole 22, and filter the liquid nitrogen through the filter assembly 23 to avoid cross-contamination between biological materials. When the vitrification freezing tool needs to be moved, set the vitrification freezing tool vertically, and store the liquid nitrogen in the connecting groove 32. The liquid nitrogen in the connecting groove 32 can enter the sleeve 21 through the through-hole 22, so that the biological materials in the sleeve 21 can be continuously maintained at a low temperature by the liquid nitrogen, playing a role in protecting the biological materials. When the vitrification freezing tool is taken out of the liquid nitrogen, the vitrification freezing tool moves in the atmospheric environment, and the liquid nitrogen in the sleeve 21 is discharged through the through-hole 22 when vaporizing, playing a role in ensuring safety during the handling of the vitrification freezing tool.

[0068] S6. Thawing: Take out the carrier rod 12 from the sleeve 21 in liquid nitrogen and quickly insert the carrier rod 12 into the thawing solution.

[0069] This embodiment has the following advantages:

[0070] After the biological materials are processed in the processing tank 13, the excess processing liquid is discharged through the drain port 16 by capillary action to achieve the minimum droplet volume. Install the sleeve 21 on the connecting body 11 and make the sleeve 21 cover the carrier rod 12, and then cryopreserve the biological materials on the carrier rod 12 connected to the vitrification freezing tool. After the biological materials are processed in the processing tank 13, there is no need to take out the biological materials from the processing tank 13. Directly freeze the vitrification freezing tool and the internal biological materials, realizing the integration of pretreatment and cryopreservation, simplifying the operation steps, facilitating the processing of biological materials. At the same time, the integration of pretreatment and cryopreservation is conducive to realizing batch and automated management of biological materials.

[0071] From the treatment of biological materials with the treatment liquid until the cryopreservation and thawing of the biological materials, there is no need to move the biological materials out of the processing tank 13, which can greatly reduce the probability of damage and contamination of biological materials during handling, playing a role in protecting biological materials. After the sleeve 21 is installed on the connecting body 11, multiple vitrification freezing tools can be transported simultaneously by handling equipment, enabling batch cryopreservation of biological materials.

[0072] By setting the sleeve 21, it can protect the biological material on the carrier rod 12, avoid contamination during the batch processing of the equipment, and ensure the reliability of the experimental process. During liquid nitrogen processing and storage, the filter component 23 can perform bidirectional filtration on the liquid nitrogen. When the liquid nitrogen enters the inside of the sleeve 21 through the through-hole 22, the filter component 23 filters the liquid nitrogen, which can prevent the pollutants in the liquid nitrogen from entering the sleeve 21 and causing contamination of the biological material, thereby preventing the biological material from being contaminated. The filter component can also filter the liquid nitrogen to prevent the liquid nitrogen outside the sleeve 21 from being contaminated, and can prevent cross-contamination of biological materials in different vitrification freezing tools.

[0073] The method can batch-add biological materials to a plurality of vitrification freezing tools, and can realize batch processing of biological materials in a plurality of vitrification freezing tools, and has the advantage of high processing efficiency.

[0074] Through the integrated operation of pretreatment and cryopreservation, the transport equipment can conveniently transport the vitrification freezing tool and the biological material inside it into liquid nitrogen for cryopreservation, which has the advantage of facilitating batch and automated management of biological materials.

[0075] A clamp matched with the connection part 31 is arranged on the transporting equipment, so as to facilitate the transport of the vitrification freezing tool through the connection part 31, facilitate connection and movement, and facilitate batch processing.

[0076] The convex strip matched with the connecting groove 32 is inserted into the connecting groove 32 , and the friction between the convex strip and the connecting groove 32 enables the convex strip to drive the connecting part 31 to move, thereby realizing the overall movement of the vitrification freezing tool driven by the connecting part 31 .

[0077] By setting the first inclined surface 17 and the second inclined surface 18, the treatment liquid and the biological material can be concentrated at the drain port 16 by gravity, which makes it convenient for the biological material to contact with the treatment liquid and to discharge the treatment liquid from the treatment tank 13 through the drain port 16.

[0078] The liquid discharge port 16 is arranged near the liquid adding tank 14 to facilitate the liquid in the liquid adding tank 14 to flow to the biological material, facilitate the contact between the biological material and the treatment liquid, and play a role in facilitating the treatment of the biological material.

[0079] Through the integrated operation of pretreatment and cryopreservation, the transport equipment can conveniently transport the vitrification freezing tool and the biological materials inside it into liquid nitrogen for cryopreservation, which is conducive to the batch and automated management of biological materials.

[0080] Embodiment 2:

[0081] refer to Figure 6, a vitrification freezing tool, which differs from Example 1 in that the inner contour of the cross section of the connecting groove 32 is overall "hexagonal".

[0082] This embodiment has the following advantages:

[0083] By adopting the connection grooves 32 of different shapes, it can be suitable for different extension rods, thus improving the applicability.

[0084] Embodiment 3:

[0085] refer to Figure 7 , a vitrification freezing tool, which differs from Example 1 in that the inner contour of the cross section of the connecting groove 32 is generally "rectangular".

[0086] This embodiment has the following advantages:

[0087] By adopting the connection grooves 32 of different shapes, it can be suitable for different extension rods, thus improving the applicability.

[0088] Embodiment 4:

[0089] refer to Figure 8 , a vitrification freezing tool, which differs from Example 1 in that the inner contour of the cross section of the connecting groove 32 is generally "triangular".

[0090] This embodiment has the following advantages:

[0091] By adopting the connection grooves 32 of different shapes, it can be suitable for different extension rods, thus improving the applicability.

[0092] Embodiment 5:

[0093] refer to Fig. 9 , a vitrification freezing tool, which differs from Example 1 in that the inner contour of the cross section of the connecting groove 32 is "elliptical" as a whole.

[0094] This embodiment has the following advantages:

[0095] By adopting the connection grooves 32 of different shapes, it can be suitable for different extension rods, thus improving the applicability.

[0096] Embodiment 6:

[0097] refer to Fig.10 , a vitrification freezing tool, which is different from Example 1 in that: a connecting portion 31 is arranged on a sleeve 21, and the connecting portion 31 is fixedly connected to the sleeve 21.

[0098] This embodiment has the following advantages:

[0099] The connecting part 31 is arranged on the sleeve 21 , and the connecting part 31 is moved, so that the sleeve 21 and the connecting body can be driven to move through the connecting part 31 , thereby realizing the function of transporting the vitrification freezing tool through the connecting part 31 .

[0100] Embodiment 7:

[0101] refer to Fig.11 , a vitrification freezing tool, which is different from Example 1 in that: the filter component 23 includes a first filter element 24.

[0102] This embodiment has the following advantages:

[0103] The filter assembly 23 realizes the function of filtering the liquid nitrogen passing through the through-hole 22 .

[0104] Embodiment 8:

[0105] refer to Fig.12 , a vitrification freezing tool, which is different from Example 1 in that the filter component 23 includes a second filter element 25.

[0106] This embodiment has the following advantages:

[0107] The filter assembly 23 realizes the function of filtering the liquid nitrogen passing through the through-hole 22 .

[0108] According to the disclosure and teaching of the above description, those skilled in the art to which the present invention belongs may also make changes and modifications to the above embodiments. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and some modifications and changes to the invention should also fall within the scope of protection of the claims of the present invention. In addition, although some specific terms are used in this specification, these terms are only for the convenience of description and do not constitute any limitation to the invention.

Claims

1. A vitrification freezing tool, comprising a connecting body (11) and a carrier rod (12), wherein the carrier rod (12) is fixedly mounted on the connecting body (11), Features: The carrier rod (12) is provided with a treatment tank (13) and a liquid addition tank (14), the carrier rod (12) is provided with a liquid guide channel (15), the two ends of the liquid guide channel (15) are respectively connected to the treatment tank (13) and the liquid addition tank (14), the connector (11) is detachably provided with a sleeve (21), the connector (11) is provided with a through-hole (22), and the connector (11) is fixedly provided with a filter assembly (23) that blocks the through-hole (22); The filter assembly (23) is made of one or more of a stainless steel filter membrane, a polymer filter membrane and a ceramic membrane, and the sleeve (21) and the carrier rod (12) are made of one or more of polyethylene, polypropylene, polystyrene, cycloolefin copolymer and acrylic; It also includes a connecting portion (31), wherein the connecting portion (31) is arranged on the connecting body (11), and the connecting portion is fixedly connected to the connecting body; The connecting portion (31) is provided with a connecting groove (32), and an extension rod (41) matching with the connecting groove (32) is installed in the connecting groove (32), and the vitrification freezing tool is driven to move by the extension rod; The filter assembly includes a first filter element and a second filter element, and the first filter element and the second filter element are respectively arranged at both ends of the through-hole; the filter assembly adopts one or more of a stainless steel filter membrane, a polymer material filter membrane and a ceramic membrane; The carrier rod (12) is provided with a liquid discharge port (16), the liquid discharge port (16) is in communication with the treatment tank (13), the position height of the liquid guide channel (15) is higher than the liquid discharge port (16), the width of the liquid discharge port (16) is set to a size capable of preventing the passage of biological materials, and the setting of the width of the liquid discharge port (16) includes one or a combination of preventing the passage of biological materials by forming a restriction by setting a width size or preventing the passage of biological materials by forming a restriction by liquid surface tension.

2. The vitrification tool according to claim 1, Features: The inner profile of the cross section of the connecting groove (32) is any one of "star-shaped", "hexagonal", "rectangular", "triangular" and "elliptical".

3. The vitrification tool according to claim 1, Features: The treatment tank (13) is provided with a first inclined surface (17) and a second inclined surface (18), and the lowest positions of the first inclined surface (17) and the second inclined surface (18) are both located at the liquid discharge port (16).

4. The vitrification tool according to claim 1, Features: The liquid discharge port (16) is located on a side of the treatment tank (13) close to the liquid addition tank (14).

5. A method for using a vitrification tool, It is characterized in that A vitrification freezing tool as claimed in claim 1 is used, the vitrification freezing tool comprising a connector (11) and a carrier rod (12), the carrier rod (12) being fixedly mounted on the connector (11), the carrier rod (12) being provided with a processing tank (13) and a liquid adding tank (14), the carrier rod (12) being provided with a liquid conducting channel (15), the two ends of the liquid conducting channel (15) being respectively connected to the processing tank (13) and the liquid adding tank (14), the connector (11) being detachably mounted with a sleeve (21), the connector (11) being provided with a through-hole (22), the connector (11) being fixedly mounted with a filter assembly (23) for blocking the through-hole (22); The method comprises the following steps: S1. Place the biological material: remove the sleeve (21) from the connector (11) and place the biological material in the treatment tank (13); S2, adding treatment liquid: adding the treatment liquid of the biological material into the liquid adding tank (14), and the treatment liquid enters the treatment tank (13) through the liquid guide channel (15) and contacts with the biological material; S3, discharging the treatment liquid: after the biological material is treated, the treatment liquid in the treatment tank (13) is discharged from the treatment tank (13) through the addition tank (14), so that the biological material remains in the treatment tank (13); S4, vitrification and sleeve installation: liquid nitrogen is placed in the sleeve (21), the carrier rod (12) containing the biological material is inserted into the liquid nitrogen in the sleeve (21), and the sleeve (21) is installed on the connector (11); S5. Cryopreservation: liquid nitrogen is allowed to enter the sleeve (21) through the through-hole (22), and the liquid nitrogen is filtered through the filter assembly (23) to avoid mutual contamination between biological materials. The liquid nitrogen in the sleeve (21) is discharged through the through-hole (22) when it is gasified.

6. The method for using the vitrification freezing tool according to claim 5, It is characterized in that In the step S1, the following steps are also included: Prepare a plurality of vitrification freezing tools, remove the sleeves (21) on the plurality of vitrification freezing tools from the connector (11), and place biological materials in a plurality of processing tanks (13) respectively; After step S1 and before step S2, the following steps are also included: S1.

1. Batch processing: Repeat step S1 multiple times to obtain multiple vitrification freezing tools containing biological materials.

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