A closed-type refrigeration support rod and its operation method

Abstract

The present application discloses a closed freezing carrier rod and an operation method. The counterweight parts supporting the carrier rod are set as two parts, and are detachably connected to the connecting part of the carrier rod through a hollow nested part. The hollowed-out part is used to increase the cooling rate of cells or embryos on the slide part of the carrier rod. The counterweight parts are used to ensure that the carrier rod sinks completely to the bottom of the liquid nitrogen bucket. Finally, the carrier rod and the counterweight parts are enclosed by a sleeve, solving the problem that compared with open freezing carriers, the cooling rate of closed carriers in the market generally becomes slower and the processing efficiency decreases.

Description

Technical Field

[0001] This application relates to the field of reproductive medicine technology, and in particular to a closed cryopreservation device and its operation method. Background Technology

[0002] Biological samples, especially tissue samples, play a crucial role in the study of various diseases. With the development of precision medicine, the importance of tissue samples has become even more prominent. Therefore, the construction of biobanks has received unprecedented attention, and tissue cryopreservation technology has become a research hotspot. Whether tissue cells can maintain good activity and function after cryopreservation is of great significance for basic, clinical, and translational medical research on diseases.

[0003] Currently, there are two main methods for tissue cryopreservation: programmed freezing and vitrification. Programmed freezing has significant drawbacks, including long processing time, formation of intracellular ice crystals, and dependence on specialized equipment. Vitrification, on the other hand, effectively avoids ice crystal formation, is simple to operate, and offers superior cryopreservation results, making it a hot topic in cryopreservation research in recent years. Vitrification was first used by Rall and Fahy in 1985, who successfully cryopreserved mouse embryos using a vitrification solution containing a mixture of multiple cryoprotectants. Since then, this method has been widely applied to the cryopreservation of various biological samples.

[0004] Human oocytes can be cryopreserved using either traditional slow freezing or vitrification. Slow freezing employs a low concentration of cryoprotectant and a slow cooling rate, achieving a balance between osmotic dehydration and temperature reduction, theoretically preventing ice crystal formation within the cells. Vitrification, on the other hand, uses a high concentration of cryoprotectant and an extremely rapid cooling rate, directly transitioning the cells from room temperature to a vitrified state, thus avoiding intracellular ice crystal formation.

[0005] Currently, there are two types of vitrification cryoprotectants: open cryoprotectants and closed cryoprotectants. Most animal research laboratories and human assisted reproductive technology centers currently use open cryoprotectants for the cryopreservation of gametes and embryos. This is because open cryoprotectants allow the solution to directly contact liquid nitrogen, achieving rapid cooling, which helps in the formation of the vitrified state and reduces ice crystal damage.

[0006] However, open cryopreservation media may lead to cross-contamination between embryos or embryo contamination due to liquid nitrogen contamination, making the use of closed cryopreservation media particularly necessary for human assisted reproductive technology centers. While closed cryopreservation media on the market offer diverse structures and can reduce or avoid damage to cells / embryos caused by ice crystal formation, they generally suffer from slower cooling rates and reduced processing efficiency compared to open cryopreservation media. Summary of the Invention

[0007] This application provides a closed-type cryogenic carrier and its operating method, which solves the problem that closed-type carriers on the market generally have slower cooling rates and reduced processing efficiency compared to open-type cryogenic carriers.

[0008] In view of this, the first aspect of this application provides a closed-type refrigeration support rod, comprising:

[0009] A sleeve, the sleeve being used to be immersed in a refrigerant;

[0010] The carrier rod includes a slide portion for placing a cryogenic carrier;

[0011] The counterweight is a hollow structure with a hollow part for accommodating the inserted carrier rod, including a hollow nesting part and a hollow part, wherein the hollow part is used to accommodate the carrier plate part of the inserted carrier rod.

[0012] Optionally, the boss at the connection end between the counterweight and the support rod is provided with an inwardly inclined surface.

[0013] Optionally, the radial cross-sectional area of ​​the hollow nested portion accounts for up to 40% of the radial cross-sectional area of ​​the counterweight.

[0014] Optionally, the axial length of the hollowed-out portion is at least 1 mm, and the axial length of the hollowed-out portion is less than the axial length of the counterweight.

[0015] Optionally, the counterweight has an inwardly inclined surface at the end away from the connection end with the support rod.

[0016] Optionally, the carrier rod further includes a connecting portion and handhold portions disposed at both ends of the connecting portion opposite to the carrier portion.

[0017] Optionally, the handle portion of the carrier rod is provided with a cutting plane parallel to the axis.

[0018] Optionally, the cutting plane of the handle portion of the carrier rod and the placement surface of the carrier plate portion of the carrier rod are at the same horizontal plane.

[0019] Optionally, a limiting member is provided on the connecting part of the carrying rod, and the radial cross-section of the connecting part with the limiting member is larger than the radial cross-section of the counterweight.

[0020] Optionally, the end of the placement surface of the carrier portion of the carrier rod is provided with an ink mark.

[0021] Optionally, the sleeve is a hollow structure with one end fully open and the other end fully closed.

[0022] Optionally, the wall thickness of the sleeve is 0.01 mm to 1.2 mm.

[0023] Optionally, the enclosed freezing carrier rod further includes an inner sleeve, which is a hollow tubular structure. The upper end of the inner sleeve is detachably connected to the connecting portion of the carrier rod, and the inner sleeve sleeves the connecting portion of the carrier rod and a part of the carrier sheet portion.

[0024] Optionally, the lower end of the inner sleeve matches the size of the hollow nesting portion to achieve a detachable connection between the inner sleeve and the counterweight.

[0025] The second aspect of the present application provides an operation method for an enclosed freezing carrier rod, including:

[0026] S1. Pass the carrier sheet portion of the carrier rod through the hollow nesting portion of the counterweight and insert it into the hollow portion of the counterweight, so as to achieve a detachable connection between the carrier rod and the counterweight;

[0027] S2. Put the carrier rod and the counterweight together into a sleeve with one end fully enclosed, and use a sealing machine to seal the open end of the sleeve;

[0028] S3. With the counterweight at the bottom, put the sleeve, the carrier rod and the counterweight together into a liquid nitrogen tank for frozen storage.

[0029] Optionally, the step S1 may be:

[0030] S11. Insert the upper end of the inner sleeve into the hollow nesting portion of the counterweight to achieve a detachable connection between the inner sleeve and the counterweight;

[0031] S12. Pass the carrier sheet portion of the carrier rod through the inner sleeve and insert it into the hollow portion of the counterweight, so that the connecting portion of the carrier rod is sleeved inside the inner sleeve and is detachably connected to the inner sleeve.

[0032] It can be seen from the above technical solutions that the embodiments of the present application have the following advantages:

[0033] In the present application, an enclosed freezing carrier rod is provided. The counterweight supporting the carrier rod is set in two parts, and is detachably connected to the connecting portion of the carrier rod through the hollow nesting portion. The hollow portion is used to increase the cooling rate of cells or embryos on the carrier sheet portion of the carrier rod. The counterweight is used to ensure that the carrier rod sinks completely to the bottom of the liquid nitrogen bucket. The ratio of the radial cross-sectional area of the hollow portion affecting the cooling rate to the wall thickness of the sleeve is restricted to further increase the cooling rate. Finally, the carrier rod and the counterweight are enclosed by the sleeve, solving the problem that the cooling rate of the enclosed carrier in the market is generally slower than that of the open freezing carrier and the processing efficiency is reduced. Description of the Drawings

[0034] Figure 1This is a schematic diagram of a closed-type refrigeration support rod in an embodiment of this application;

[0035] Figure 2 This is a schematic diagram of the structure of the carrier rod in the embodiments of this application;

[0036] Figure 3 This is a schematic diagram of the counterweight structure in an embodiment of this application;

[0037] Figure 4 This is a partially enlarged structural diagram of the counterweight in an embodiment of this application;

[0038] Figure 5 This is a schematic diagram of a closed-type refrigeration carrier rod with an inner sleeve in an embodiment of this application;

[0039] Figure 6 This is a schematic diagram of the model structure of the cryogenic support rod in the embodiments of this application;

[0040] Figure 7 This is a comparison diagram of simulation and experimental results of PVC sleeves in the embodiments of this application;

[0041] Figure 8 This application provides a comparison of simulation and experimental results for the PE sleeve in the embodiments.

[0042] Figure 9 This is a schematic diagram showing the cooling-temperature vs. time of each product in the embodiments of this application compared to existing design models;

[0043] The attached figures are labeled as follows:

[0044] 1. Support rod; 2. Counterweight; 3. Sleeve; 4. Inner sleeve; 11. Connecting part; 12. Handheld part; 13. Carrier part; 14. Limiting part; 15. Ink marking; 21. Hollow nested part; 22. Hollowed-out part; 23. Solid part; 24. Supporting part. Detailed Implementation

[0045] To enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present application, and not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present application.

[0046] This application designs a closed-type cryogenic carrier and its operation method, which solves the problem that closed-type carriers on the market generally have slower cooling rates and reduced processing efficiency compared to open-type cryogenic carriers.

[0047] In some embodiments, the cells may be cells and / or multicellular bodies derived from oocytes, zygotes, blastocysts, and embryos of humans, non-human primates, dogs, cattle, horses, pigs, sheep, goats, cats, buffalo, guinea pigs, hamsters, rabbits, rats, and mice.

[0048] For easier understanding, please refer to Figures 1 to 4 ,in Figure 1 This is a schematic diagram of a closed-type refrigeration support rod in an embodiment of this application, as shown below. Figure 1 As shown, specifically:

[0049] Sleeve 3, sleeve 3 is used to be immersed in refrigerant;

[0050] Carrier rod 1, carrier rod 1 includes a slide portion 13 for placing a freezing carrier;

[0051] The counterweight 2 has a hollow structure that accommodates the inserted support rod 1, including a hollow nesting part 21 and a hollow part 22. The hollow part 22 is used to accommodate the support plate part 13 of the inserted support rod 1.

[0052] It should be noted that the sleeve 3 is a hollow structure with one end fully open and the other end fully closed. The material of the sleeve 3 has the characteristics of biocompatibility, resistance to liquid nitrogen, radiation resistance and thermoplasticity. The closed end of the hollow part 22 near the counterweight 2 is usually pre-sealed, and the other end is sealed by the operator after placing the cells or embryos.

[0053] The seal remains intact for extended periods in a liquid nitrogen environment, exhibiting excellent sealing performance. Sealing methods include conventional methods such as ultrasonic sealing, high-frequency sealing, heat sealing, and infrared sealing.

[0054] The sealing shape can be a conventional shape such as a fan, square, or circle, and the pattern can be a variety of patterns such as flat patterns, grid patterns, or wavy patterns.

[0055] Sleeve 3 is a transparent tube, providing the operator with a visual view of the operation and reducing operational errors.

[0056] The counterweight 2 has a hollow structure for accommodating the inserted support rod 1, including a hollow nesting part 21 and a hollow part 22. The hollow part 22 is used to accommodate the support plate part 13 of the inserted support rod 1. One end of the counterweight 2 is open, so that the support plate part 1 of the support rod 1 is inserted through the opening of the counterweight 2 and passes through the hollow nesting part 21 of the counterweight 2 into the hollow part 22.

[0057] Since the carrier rod 1's buoyancy in liquid nitrogen could lead to the loss of cells or embryos in the slide portion 13, a minimum weight for the counterweight 2 was determined to ensure the carrier rod 1 sinks completely to the bottom of the liquid nitrogen container. This was achieved by calculating factors such as the carrier rod 1's buoyancy, volume, liquid nitrogen buoyancy, and weight. Since the weight is fixed, stainless steel can be used to increase density, but this would reduce the volume of the counterweight 2.

[0058] The counterweight 2 can be made of metal to further accelerate heat conduction.

[0059] The shape of the hollow part 22 of the counterweight 2 can be a conventional shape such as a fan, a square, or a circle, and is not limited here.

[0060] Furthermore, the boss at the connection end between the counterweight 2 and the support rod 1 is provided with an inward slope.

[0061] It should be noted that the boss at the connection end between the counterweight 2 and the carrier rod 1 is provided with an inward slope, which can prevent the carrier plate part 13 of the carrier rod 1 from touching the interior of the counterweight 2, and further limit the connection between the carrier rod 1 and the counterweight 2.

[0062] Furthermore, the radial cross-sectional area of ​​the hollow nested part 21 accounts for at most 40% of the radial cross-sectional area of ​​the counterweight 2.

[0063] It should be noted that thermodynamic simulations revealed that the key factor affecting the cooling rate of the support rod 1 includes the proportion of the radial cross-sectional area of ​​the hollow nested portion 21 of the counterweight 2 to the total radial cross-sectional area of ​​the counterweight 2. Through simulation comparisons of multiple transversely hollowed-out models, multiple longitudinally hollowed-out models, models with different metal wall thicknesses, and models with different materials, it was confirmed that the radial cross-sectional area of ​​the hollow nested portion 21 accounts for at most 40% of the radial cross-sectional area of ​​the counterweight 2.

[0064] Optimally, the structural design in which the radial cross-sectional area of ​​the hollow nested part 21 of the counterweight 2 accounts for 17% of the radial cross-sectional area of ​​the counterweight 2 can optimally improve the cooling rate.

[0065] It is understandable that, since the hollow nested part 21 is a hollow structure, it contains a hollow portion. The radial cross-sectional area of ​​the hollow nested part 21 of the counterweight 2 refers to the area of ​​the hollow nested part 21 after deducting the hollow and openwork portions from its radial cross-sectional area; the radial cross-sectional area of ​​the counterweight 2 refers to the area of ​​the annular portion of the counterweight 2 after deducting the hollow portion from its radial cross-sectional area. That is, the radial cross-sectional area of ​​the counterweight 2 is equal to the sum of the radial cross-sectional area of ​​the hollow nested part 21 and the radial cross-sectional area of ​​the openwork part 22. Furthermore, the axial length of the openwork part 22 is at least 1 mm, and the axial length of the openwork part 22 is less than the axial length of the counterweight 2.

[0066] It should be noted that the hollow part 22 is a channel opened on the wall of the counterweight 2 to achieve the hollowing. The hollow part 22 corresponds to the position of the carrier plate 13 after the carrier rod 1 is inserted into the counterweight 2. The axial length of the channel of the hollow part 22 is at least 1mm, which can cover almost the entire counterweight 2. However, in order to ensure the existence of the hollow nesting part 21 and to avoid the carrier rod 1 being unable to connect with the counterweight 2, the axial length of the hollow part 22 is less than the axial length of the counterweight 2.

[0067] The perforation of the perforated part 22 preferably corresponds to the placement surface of the plate part 13 of the carrier rod 1, so as to improve the cooling rate.

[0068] Furthermore, the end of the counterweight 2 away from the end connected to the support rod 1 is provided with an inward slope.

[0069] It should be noted that, after being frozen by liquid nitrogen, if the ramp is missing, it will be difficult to pull the counterweight 2 and the support rod 1 out of the sleeve 3. The end of the counterweight 2 away from the end connected to the support rod 1 is also provided with an inward ramp, which makes it easier to insert and remove the counterweight 2 and the support rod 1 from the sleeve 3.

[0070] Furthermore, the carrier rod 1 also includes a connecting part 11 and handhold parts 12 disposed at both ends of the connecting part 11 opposite to the carrier part 13.

[0071] It should be noted that the carrier rod 1 includes a connecting part 11 and a hand-held part 12 disposed at both ends of the connecting part 11 opposite to the carrier part 13. The hand-held part 12 is used for operation by the operator, and the connecting part 11 is used to detachably connect the carrier rod 1 and the counterweight 2.

[0072] Furthermore, the handle portion 12 of the carrier rod 1 is provided with a cutting plane parallel to the axis.

[0073] It should be noted that the handle 12 of the carrier rod 1 can be provided with a cut plane parallel to the axis, allowing the operator to distinguish the front and back sides by printing graphic or text markings on the handle 12. Simultaneously, the flat design also allows the operator to record patient information using a cryogenic marker. Compared to using affixed barcodes, using a cryogenic marker to record information avoids the risk of barcodes detaching during long-term storage in a liquid nitrogen environment.

[0074] Furthermore, the cutting plane of the handle portion 12 of the carrier rod 1 and the placement surface of the plate portion 13 of the carrier rod 1 are at the same horizontal plane.

[0075] It should be noted that the cutting plane of the hand-held part 12 of the carrier rod 1 and the placement surface of the plate part 13 of the carrier rod 1 are on the same horizontal plane, which makes it easier to distinguish the front and back sides and to directly observe the information.

[0076] Furthermore, a limiting member 14 is provided on the connecting part 11 of the load bar 1, and the radial cross section of the connecting part 11 with the limiting member 14 is larger than the radial cross section of the counterweight 2.

[0077] It should be noted that a limiting member 14 can be provided on the connecting part 11 of the carrier rod 1. The shape of the limiting member 14 can be arbitrary, but the radial cross section of the connecting part 11 with the limiting member 14 is larger than the radial cross section of the counterweight 2. This ensures that the connection between the carrier rod 1 and the counterweight 2 is not too tight and difficult to insert or remove, nor is it too loose and causes the carrier rod 1 to fall off.

[0078] Furthermore, an ink mark 15 is provided at the end of the placement surface of the carrier portion 13 of the carrier rod 1.

[0079] It should be noted that by setting an ink mark 15 at the end of the placement surface of the slide section 13, when vitrifying and freezing cells, the operator can quickly locate the cell position in the slide section 13 by identifying the black ink under a microscope.

[0080] Furthermore, the sleeve 3 is a hollow structure with one end fully open and the other end fully closed.

[0081] Furthermore, the wall thickness of sleeve 3 is 0.01mm to 1.2mm.

[0082] It should be noted that, based on the construction of the simulation model, the wall thickness of the sleeve 3 has a significant impact on the cooling rate, preferably ranging from 0.01 mm to 1.2 mm, with the most preferred value being 0.18 mm.

[0083] Furthermore, the counterweight 2 may also include a solid part 23, which is used to provide downward force for the counterweight 2.

[0084] It should be noted that the solid part 23 of the counterweight 2 is located at the end away from the connection end between the counterweight 2 and the support rod 1.

[0085] Furthermore, such as Figure 5 As shown, the closed-type cryogenic carrier provided in this application embodiment also includes an inner sleeve 4. The inner sleeve 4 is a hollow tube structure. The upper end of the inner sleeve 4 is detachably connected to the connecting part 11 of the carrier 1. The inner sleeve 4 covers the connecting part of the carrier 1 and a portion of the carrier plate part.

[0086] It should be noted that the inner tube 4 is a transparent hollow tube made of a relatively hard material. Since the outer tube 3 needs to be cut open to remove the carrier rod 1 after freezing, the hardness of the inner tube 4 ensures that scissors cannot cut it, thus protecting the cells from damage.

[0087] The inner sleeve 4 houses the connecting part 11 of the carrier rod 1 and the carrier piece part 13 of the preset size. Optimally, the inner sleeve 4 houses one-third of the carrier piece part 13 of the carrier rod 1, so that two-thirds of the carrier piece part 13 of the carrier rod 1 is inserted into the counterweight 2.

[0088] Furthermore, the lower end of the inner sleeve 4 is matched with the size of the counterweight 2 to achieve a detachable connection between the inner sleeve 4 and the counterweight 2.

[0089] It should be noted that the connection between the inner sleeve 4 and the counterweight 2 is such that the inner sleeve 4 is inside and the counterweight 2 is outside, which ensures that the carrier plate part 13 of the carrier rod 1 will not touch the inner wall of the counterweight 2 no matter how it is inserted into the counterweight 2, thus providing good operability.

[0090] Furthermore, the upper end of the inner sleeve 4 is provided with an ink mark to indicate the correct insertion direction for the carrier portion 13 of the carrier rod 1.

[0091] This application's embodiments explore, through the establishment of a simulation model, the optimal radial cross-sectional area ratio of the hollowed-out portion of the counterweight, the sleeve wall thickness, and the sleeve inner diameter under the structure of the closed-type cryogenic support rod provided in this application, as detailed below:

[0092] The structural models of products A, B, C, and D are as follows: Figure 6 As shown in the figure. Their sleeve wall thickness is 0.28 mm, the same as the existing design. Product E has a sleeve wall thickness of 0.18 mm, while its other structures are the same as Product D. Detailed parameters are described in Table 1. Products F and G are sleeve models with different materials, wall thicknesses, and outer diameters, used to compare simulation and experimental results.

[0093] Table 1. Introduction to the Simulation Model

[0094]

[0095]

[0096] The simulation conditions are set as follows:

[0097] The simulation uses a solid-liquid two-phase flow model, and the turbulence model uses the Laminar model for unsteady-state solution. The model time step is 0.1s, and the calculation time is 15-25s.

[0098] The acceleration due to gravity is 9.81 m / s². 2 The air inside the refrigerated support rod is assumed to be incompressible-ideal gas, with an operating pressure of 101325 Pa, a temperature of 25 °C, and a density of 1.225 kg / m³. 3 .

[0099] Assuming the cryogenic support rod is covered by liquid nitrogen and the surface temperature of the outer casing is -196°C;

[0100] The cell model was frozen at 37°C, while the rest of the environment was kept at room temperature (25°C).

[0101] The simulation model uses a Coupled mode for calculation, a Body Force Weighted pressure space discretization scheme, and other settings are configured as a second-order upwind mode.

[0102] Cryotherapy scenario: Cells at 37°C are attached to a slide on a closed carrier rod and rapidly immersed in liquid nitrogen at -196°C in a room temperature environment of 25°C for cooling. The liquid nitrogen cannot penetrate into the sleeve.

[0103] The simulation results obtained are as follows:

[0104] The simulation and measured results for products F and G are as follows: Figure 7 and 8 As shown, the simulation results for PVC and PE sleeves are not significantly different from the measured values. In the optimized simulation model, the air density changes with temperature during the calculation process. Considering the influence of gravity, the convection effect during the cooling process of the air inside the refrigerated support rod will be more pronounced. The old method ignored the density change of the air inside the support rod during cooling, and the convection effect in the airflow domain inside the refrigerated support rod was not effectively calculated (the velocity field of the internal airflow domain is 0). Therefore, the heat transfer simulation calculation of the old method may have a larger error, while the optimized simulation calculation method is more realistic.

[0105] Therefore, according to Figure 9 The conclusions drawn from the cooling-temperature vs. time diagrams of various products compared to existing design models and measured values ​​are as follows:

[0106] 1. Under the same wall thickness, the inner diameter of the sleeve has little effect on the cooling rate, which is consistent with the simulation and test results.

[0107] 2. The sleeve wall thickness has a significant impact on the cooling rate, which is consistent with the simulation and testing results.

[0108] 3. The simulation and measured results for PVC and PE sleeves are not significantly different.

[0109] We know that the time required to cool to -130°C is crucial for cells; the shorter the time, the less damage to the cells on the slide. According to... Figure 8 As shown, in some embodiments, compared with the measured values ​​of the existing design, products B, C, D and E all perform better. Correspondingly, the optimal range of the sleeve wall thickness is 0.18mm to 0.28mm, and the optimal range of the radial cross-sectional area of ​​the hollow part of the counterweight is 80% to 85%.

[0110] Further analysis and comparison show that, with the same sleeve wall thickness, the larger the radial cross-sectional area of ​​the hollow part of the counterweight, the less cooling time is required and the higher the cooling rate is during the process of cooling down to -130℃.

[0111] Further analysis and comparison show that, when the radial cross-sectional area of ​​the hollow part of the counterweight is the same, the smaller the wall thickness of the sleeve, the less cooling time is required and the higher the cooling rate is during the process of cooling down to -130℃.

[0112] Therefore, under conditions superior to existing designs, the radial cross-sectional area of ​​the hollow portion of the counterweight, ranging from 65% to 85%, is inversely proportional to the wall thickness of the sleeve, ranging from 0.18 mm to 1.2 mm. That is, the larger the radial cross-sectional area of ​​the hollow portion and the relatively smaller the sleeve wall thickness, the higher the cooling rate. In the closed-loop refrigeration support rod provided in this application, the sleeve wall thickness is 0.18 mm, and the radial cross-sectional area of ​​the hollow portion of the counterweight is 83%, meaning the radial cross-sectional area of ​​the hollow nested portion of the counterweight is 17%, which represents the optimal model data.

[0113] This application also provides a method for operating a closed-loop refrigeration support rod, including:

[0114] S1. Pass the carrier plate portion of the carrier rod through the hollow nested portion of the counterweight and insert it into the hollow portion of the counterweight to make the carrier rod and the counterweight detachable.

[0115] S2. Place the support rod and the counterweight together into a sleeve that is fully enclosed at one end, and seal the open end of the sleeve using a sealing machine.

[0116] S3. Place the counterweight at the bottom, and put the sleeve, the support rod, and the counterweight together into a liquid nitrogen tank for freezing and preservation.

[0117] Furthermore, in the operation method of the aforementioned enclosed cryogenic support rod, step S1 may be:

[0118] S11. Insert the upper end of the inner sleeve into the hollow nested part of the counterweight to make the inner sleeve and the counterweight detachable.

[0119] S12. Insert the carrier plate portion of the carrier rod through the inner sleeve into the hollow portion of the counterweight, so that the connecting portion of the carrier rod is fitted inside the inner sleeve and detachably connected to the inner sleeve.

[0120] Specifically, in the freezing scenario: The operator takes out the carrier rod from the packaging bag, checks that its surface has no scratches, stains or burrs, holds the holding part of the carrier rod with the hand, turns the side with printed graphics or text marks (i.e., the radial cross-section) upwards and records the patient information (stick a liquid nitrogen-resistant label or write with a liquid nitrogen-resistant marker pen), places the slide part under the microscope, finds the ink-marked end of the slide part under the light source, places the embryo / oocyte that has been dehydrated by cryoprotectant treatment in advance at 1-2 mm in front of the ink mark, then sucks out the excess cryoprotectant around the embryo / oocyte with a pipette, inserts the slide carrying the embryo / oocyte parallel into the counterweight, and then puts them together into a sleeve with one end sealed, seals the other end with a sealing machine, and then puts them into a bucket and a liquid nitrogen tank for cryopreservation.

[0121] In the thawing scenario: Select the liquid nitrogen tank and bucket used for freezing, find the carrier rod with the patient information recorded and check it, pull out the carrier rod, hold the carrier rod with pliers in the left hand, and cut the handle part with an open scissors in the right hand. At this time, the seal of the sleeve is opened, and the operator uses tweezers in the right hand to take out the carrier rod for thawing and observe its thawing state.

[0122] In the embodiment of the present application, a closed freezing carrier rod is provided. The counterweight matching the carrier rod is set as two parts, and is detachably connected to the connecting part of the carrier rod through the hollow nested part. The hollow part is used to improve the cooling rate of the cells or embryos on the slide part of the carrier rod. The counterweight is used to ensure that the carrier rod sinks completely to the bottom of the liquid nitrogen bucket. Finally, the carrier rod and the counterweight are enclosed by the sleeve, which solves the problem that the cooling rate of the closed carriers in the market is generally slower and the processing efficiency is reduced compared with the open freezing carriers.

[0123] Those skilled in the art can clearly understand that for the convenience and brevity of description, the specific working processes of the systems, devices and units described above can refer to the corresponding processes in the foregoing method embodiments, and will not be repeated here.

[0124] The terms "first", "second", "third", "fourth", etc. (if any) in the specification of the present application and the above-mentioned drawings are used to distinguish similar objects, and do not have to be used to describe a specific order or sequence. It should be understood that such data can be interchanged under appropriate circumstances, so that the embodiments of the present application described here can be implemented in an order different from those illustrated or described here. In addition, the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device comprising a series of steps or units does not have to be limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0125] It should be understood that in this application, "at least one (item)" means one or more, and "more than" means two or more. "And / or" is used to describe the relationship between related objects, indicating that three relationships can exist. For example, "A and / or B" can represent three cases: only A exists, only B exists, and both A and B exist simultaneously, where A and B can be singular or plural. The character " / " generally indicates that the preceding and following related objects are in an "or" relationship. "At least one (item) of the following" or similar expressions refer to any combination of these items, including any combination of single or plural items. For example, at least one (item) of a, b, or c can represent: a, b, c, "a and b", "a and c", "b and c", or "a and b and c", where a, b, and c can be single or multiple.

[0126] The above-described embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A closed-type refrigeration support rod, characterized in that, include: A sleeve, the sleeve being used to be immersed in a refrigerant; The carrier rod includes a slide portion for placing a cryogenic carrier; The counterweight is a hollow structure with a hollow part for accommodating the inserted carrier rod, including a hollow nesting part and a hollow part, wherein the hollow part is used to accommodate the carrier plate part of the inserted carrier rod; The radial cross-sectional area of ​​the hollow nested part accounts for at most 15% to 35% of the radial cross-sectional area of ​​the counterweight; The boss at the connection end between the counterweight and the support rod has an inwardly inclined surface.

2. The enclosed refrigeration support rod according to claim 1, characterized in that, The axial length of the hollowed-out portion is at least 1 mm, and the axial length of the hollowed-out portion is less than the axial length of the counterweight.

3. The enclosed refrigeration support rod according to claim 1, characterized in that, The counterweight has an inwardly inclined surface at the end away from the connection with the support rod.

4. The enclosed refrigeration support rod according to claim 1, characterized in that, The carrier rod also includes a connecting part and handholds disposed at both ends of the connecting part opposite to the carrier part.

5. The enclosed refrigeration support rod according to claim 1, characterized in that, The handle portion of the carrier rod is provided with a cutting plane parallel to the axis.

6. The enclosed refrigeration support rod according to claim 5, characterized in that, The cutting plane of the handle portion of the carrier rod and the placement surface of the carrier plate portion of the carrier rod are at the same horizontal plane.

7. The enclosed refrigeration support rod according to claim 1, characterized in that, A limiting member is provided on the connecting part of the carrying rod, and the radial cross-section of the connecting part with the limiting member is larger than the radial cross-section of the counterweight.

8. The enclosed refrigeration support rod according to claim 1, characterized in that, The end of the placement surface of the carrier plate of the carrier rod is provided with an ink mark.

9. The enclosed refrigeration support rod according to claim 1, characterized in that, The sleeve is a hollow structure with one end fully open and the other end fully closed.

10. The enclosed refrigeration support rod according to claim 1, characterized in that, The wall thickness of the sleeve is 0.01mm to 1.2mm.

11. The enclosed refrigeration support rod according to claim 1, characterized in that, The enclosed cryogenic carrier rod also includes an inner sleeve, which is a hollow tube structure. The upper end of the inner sleeve is detachably connected to the connecting part of the carrier rod, and the inner sleeve covers the connecting part of the carrier rod and a portion of the carrier plate.

12. The enclosed refrigeration support rod according to claim 11, characterized in that, The lower end of the inner sleeve matches the size of the hollow nested part of the counterweight to achieve a detachable connection between the inner sleeve and the counterweight.

13. A method for operating a closed-loop refrigeration support rod, characterized in that, include: S1. Pass the carrier plate portion of the carrier rod through the hollow nested portion of the counterweight and insert it into the hollow portion of the counterweight to make the carrier rod and the counterweight detachable. S2. Place the support rod and the counterweight together into a sleeve that is fully enclosed at one end, and seal the open end of the sleeve using a sealing machine. S3. Place the counterweight at the bottom, and put the sleeve, the support rod, and the counterweight together into a liquid nitrogen tank for freezing and preservation.

14. The operating method of a closed-type refrigeration support rod according to claim 13, characterized in that, Step S1 is: S11. Insert the upper end of the inner sleeve into the hollow nested part of the counterweight, so that the inner sleeve and the counterweight are detachably connected. S12. Insert the carrier plate portion of the carrier rod through the inner sleeve into the hollow portion of the counterweight, so that the connecting portion of the carrier rod is fitted inside the inner sleeve and detachably connected to the inner sleeve.

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