Cell vitrification cryopreservation device

Abstract

The utility model relates to the technical field of low-temperature biomedicine, in particular to a cell vitrification cryopreservation device, which comprises a charging system, a cryopreservation system and a control system, in the pulsating heat pipe system, each layer of sub pulsating heat pipe comprises a condensation end, a cell container and an evaporation end; a first capillary tube is arranged in the condensation end; the first capillary tube is communicated with the filling system; a channel is arranged in the cell container, and a second capillary tube is arranged in the evaporation end; the first capillary tube, the channel and the second capillary tube are communicated in sequence to form a pulsating heat pipe loop; a cell solution is contained in the cell container, and the cell solution and the channel are arranged separately; a liquid nitrogen pool is arranged in the pre-cooling system; the condensation end is in contact with the liquid nitrogen pool, and the evaporation end is insulated from the liquid nitrogen pool. According to the utility model, liquid nitrogen is used as a cold source, and the multiple layers of sub pulsating heat pipes are used for carrying out dry-type cold conduction, so that the Leidenfrost effect is avoided, the cooling rate of a cell solution is greatly improved, and batch cryopreservation of cell samples can be realized by coupling a single charging system.

Description

Technical Field

[0001] The utility model relates to the technical field of cryobiomedicine, in particular to a cell vitrification cryopreservation device. Background Art

[0002] The cryopreservation technology of biological materials such as tissues and cells plays a key role in biomedical research, disease treatment, human fertility and species protection. The cryopreservation of biological materials is to cool the biological materials and then store them at a low temperature to reduce or suspend their physico-chemical and metabolic activities, and then restore them to the physiological state by rewarming when needed. The most important evaluation index of cryopreservation effect is the cell survival rate. During the cooling and rewarming processes, cell damage should be minimized as much as possible to improve the cell survival rate. In cryopreservation, cell damage is mainly caused by solution effects and intracellular ice formation. To reduce cell damage caused by the above mechanisms and improve the survival rate, one or more cryoprotective agents (CPAs) are usually added to the biological material samples.

[0003] There are two commonly used methods for cryopreserving biological materials: slow freezing and vitrification freezing (ultra-rapid cryopreservation). At a lower cooling rate, both solution effects and intracellular ice formation are significant. Under the influence of these two damage mechanisms, different types of cells have different optimal cooling rates, and their survival rate-cooling rate curve is in an inverted U shape. When cryopreserving multicellular biological materials, high-concentration CPA is usually used to broaden the optimal cooling rate range of each type of cell. However, the higher the CPA concentration, the stronger its chemical toxicity. Therefore, when cryopreserving multicellular biological materials by slow freezing, complex pre-freezing CPA gradient permeation and post-rewarming CPA removal operations are required to reduce the impact of chemical toxicity. When the cooling rate is high enough, the solutes inside the cells can be quickly transformed into a glassy state before the ice crystal nuclei grow.

[0004] The vitrification cryopreservation technology can theoretically minimize the damage caused by intracellular ice formation. In vitrification cryopreservation, rapid cooling and rewarming are very important. The cooling rate should be higher than the critical cooling rate to achieve vitrification of the solutions inside and outside the tissues and cells. The warming rate should be higher than the critical warming rate to prevent ice crystal regeneration during the rewarming process. Theories and experiments show that the higher the cooling and rewarming rates, the lower the required CPA concentration. Due to insufficient cooling and rewarming rates, traditional vitrification cryopreservation methods still need to use high-concentration CPA, so complex pre-freezing CPA gradient permeation and post-rewarming CPA removal operations are also required.

[0005] To perform vitrification freezing with a lower concentration of CPA, extremely high cooling and rewarming rates need to be achieved. However, the cooling rate of the existing cell vitrification cryopreservation devices is insufficient. Summary of the Utility Model

[0006] The present utility model provides a cell vitrification cryopreservation device, which is used to solve the defect of slow cooling rate in the prior art, realize the batch processing of multiple groups of cell solution samples, and utilize the advantages of the pulsating heat pipe itself, such as outstanding heat transfer capacity, no power consumption, and simple structure, to achieve high-rate cooling of the system.

[0007] The present utility model provides a cell vitrification cryopreservation device, comprising:

[0008] A filling system for providing nitrogen;

[0009] A pulsating heat pipe system, including multiple layers of sub-pulsating heat pipes, and each layer of the sub-pulsating heat pipe includes: a condensation end, a cell container, and an evaporation end; a first capillary is provided in the condensation end; the first capillary is communicated with the filling system; a channel is arranged inside the cell container, and a second capillary is provided in the evaporation end; and the first capillary, the channel, and the second capillary are communicated in sequence to form a pulsating heat pipe loop; the cell container contains a cell solution, and the cell container and the cell solution are separately arranged from the channel;

[0010] A precooling system with a liquid nitrogen pool built in; and the condensation end is in contact with the liquid nitrogen pool, and the evaporation end is insulated from the liquid nitrogen pool.

[0011] According to the cell vitrification cryopreservation device provided by the present utility model, the filling system includes:

[0012] A nitrogen cylinder for providing nitrogen with a purity of 99.999%;

[0013] A gas pipeline, one end of which is communicated with the nitrogen cylinder;

[0014] A stop valve arranged on the gas pipeline;

[0015] A filling pipe, one end of which is communicated with the other end of the gas pipeline, and the other end of the filling pipe is communicated with the first capillary.

[0016] According to the cell vitrification cryopreservation device provided by the present utility model, it further includes: a multi-channel connector for communicating the filling system and the sub-pulsating heat pipe, and the multi-channel connector includes:

[0017] A first connection section connected to the filling pipe;

[0018] A second connection section with multiple horizontal ones, and the second connection section is vertically connected to the first connection section; the second connection section is used to communicate with the first capillary.

[0019] According to the cell vitrification cryopreservation device provided by the present utility model, the filling system further includes:

[0020] A buffer tank is provided on the gas pipeline;

[0021] A molecular pump unit is connected in parallel with the nitrogen gas cylinder;

[0022] A first valve is provided between the molecular pump unit and the gas pipeline;

[0023] A second valve is provided between the nitrogen gas cylinder and the gas pipeline.

[0024] According to a cell vitrification cryopreservation device provided by the present invention, the filling system further includes:

[0025] A first pressure sensor is provided at one end of the gas pipeline close to the filling tube for detecting the pressure fluctuation at the condensation end;

[0026] A second pressure sensor is communicated with the buffer tank for detecting the pressure of the buffer tank to adjust the liquid filling rate.

[0027] According to a cell vitrification cryopreservation device provided by the present invention, the second capillary tubes are all U-shaped, the first capillary tube includes a U-shaped tube and an L-shaped tube, and there are two L-shaped tubes, which are respectively located on both sides of the condensation end, there are multiple U-shaped tubes, and multiple U-shaped tubes are all located between the two L-shaped tubes, and the L-shaped tube 2112 is communicated with the filling system.

[0028] According to a cell vitrification cryopreservation device provided by the present invention, the condensation end, the cell container and the evaporation end are all made of oxygen-free copper plates.

[0029] According to a cell vitrification cryopreservation device provided by the present invention, a heating film is provided on the back of the evaporation end.

[0030] According to a cell vitrification cryopreservation device provided by the present invention, the peripheries of the cell container and the evaporation end are wrapped with a foam layer for heat insulation and anti-pollution.

[0031] According to a cell vitrification cryopreservation device provided by the present invention, the gas pipeline and the filling tube both include stainless steel tubes.

[0032] The cell vitrification cryopreservation device provided by the present invention uses liquid nitrogen as a cold source and multi-layer sub-pulsating heat pipes for dry heat conduction. On the one hand, it avoids the Leidenfrost effect caused by direct cooling of cell solutions by liquid nitrogen at present, and greatly improves the cooling rate of cell solutions. On the other hand, aiming at the problem that the traditional pulsating heat pipe structure cannot realize batch processing of samples for cell vitrification cryopreservation, a new pulsating heat pipe structure is proposed, and batch cryopreservation of cell samples can be realized by coupling a single liquid filling system. Description of the Drawings

[0033] To more clearly illustrate the technical solutions in the present utility model or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present utility model. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0034] Figure 1 It is a schematic structural diagram of the cell vitrification cryopreservation device provided by the present utility model.

[0035] Figure 2 It is a schematic structural diagram of the pulsating heat pipe system provided by the present utility model.

[0036] Figure 3 It is a schematic structural diagram of the sub-pulsating heat pipe provided by the present utility model.

[0037] Figure 4 It is a schematic structural diagram of the filling system provided by the present utility model.

[0038] Figure 5 It is a schematic structural diagram of the multi-channel connector provided by the present utility model.

[0039] Reference numerals:

[0040] 1. Filling system; 11. Nitrogen cylinder; 12. Gas pipeline; 13. Stop valve; 14. Filling pipe; 15. Buffer tank; 16. Molecular pump unit; 17. First valve; 18. Second valve; 19. First pressure sensor; 110. Second pressure sensor; 2. Pulsating heat pipe system; 21. Condensing end; 22. Cell container; 23. Evaporating end; 24. Cell solution; 25. Cover plate; 211. First capillary; 2111. U-shaped tube; 2112. L-shaped tube; 231. Second capillary; 232. Heating film; 3. Pre-cooling system; 31. Liquid nitrogen pool; 32. Foam layer; 33. Support structure; 34. Dewar; 35. Inlet pipe; 4. Multi-channel connector; 41. First connection section; 42. Second connection section; 43. Flow channel; 44. Chamfer. Detailed implementation manners

[0041] To make the objectives, technical solutions and advantages of the present utility model clearer, the following will clearly and completely describe the technical solutions in the present utility model with reference to the drawings in the present utility model. Obviously, the described embodiments are some but not all of the embodiments of the present utility model. Based on the embodiments in the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present utility model.

[0042] In the description of the embodiments of the present utility model, it should be noted that the orientation or positional relationship indicated by the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the embodiments of the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation to the embodiments of the present utility model. In addition, the terms "first", "second", "third" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance.

[0043] The following will describe the cell vitrification cryopreservation device of the present utility model in conjunction with Figures 1 - 5 Describe the cell vitrification cryopreservation device of the present utility model.

[0044] As Figure 1 、 Figure 2 And Figure 3 shown, the present utility model provides a cell vitrification cryopreservation device, which includes a filling system 1, a pulsating heat pipe system 2 and a precooling system 3.

[0045] Among them, the filling system 1 is used to provide nitrogen. The pulsating heat pipe system 2 includes multiple layers of sub-pulsating heat pipes, and the structure of each layer of sub-pulsating heat pipes is the same. As Figure 2 shown, the sub-pulsating heat pipes can be set to five groups.

[0046] As Figure 2 shown, each layer of sub-pulsating heat pipes includes: a condensation end 21, a cell container 22 and an evaporation end 23; a first capillary 211 is provided in the condensation end 21; the first capillary 211 is communicated with the filling system 1; a channel is provided inside the cell container 22, and a second capillary 231 is provided in the evaporation end 23; and the first capillary 211, the channel and the second capillary 231 are communicated in sequence and form a pulsating heat pipe loop; a cell solution 24 is accommodated in the cell container 22, and the cell solution 24 is separated from the channel.

[0047] Among them, the first capillary 211 and the second capillary 231 can adopt stainless steel capillary segments, and the inner diameter of the stainless steel capillary segments is 1 mm, which is smaller than the critical pipe diameter specified by the liquid nitrogen PHP (Pulsating heat pipe).

[0048] Specifically, the structures of the condensation end 21 and the evaporation end 23 are exactly the same, and 10 parallel grooves with a width slightly larger than the outer diameter of the capillary segment are milled on them, and the parallel grooves are used to place the first capillary 211 and the second capillary 231.

[0049] The precooling system 3 is internally provided with a liquid nitrogen pool 31; and the condensation end 21 is in contact with the liquid nitrogen pool 31, and the evaporation end 23 is adiabatic to the liquid nitrogen pool 31.

[0050] The cell vitrification cryopreservation device provided by the present utility model, since the first capillary 211, the channel and the second capillary 231 are connected in sequence to form a pulsating heat pipe loop, the first capillary 211 is connected to the filling system 1, and the filling system 1 provides nitrogen. The nitrogen is quickly liquefied by precooling at the condensation end 21 to form alternately distributed gas-liquid plugs, and driven by the heat input at the evaporation end 23, it rapidly flows in the entire pulsating heat pipe loop. When the high-speed moving gas-liquid plugs flow through the internal channel of the cell container 22, they exchange heat violently with the cell solution on the upper surface of the container, realizing the vitrification cryopreservation of the cell solution. After the cell solution is cooled, it is transferred to liquid nitrogen for storage.

[0051] By using liquid nitrogen as the cold source and multiple-layer sub-pulsating heat pipes for dry conduction cooling, on the one hand, it avoids the Leidenfrost effect caused by the direct cooling of the cell solution by liquid nitrogen at present, and greatly improves the cooling rate of the cell solution. On the other hand, aiming at the problem that the traditional structure pulsating heat pipe cannot realize batch processing of samples for cell vitrification cryopreservation, a new pulsating heat pipe structure is proposed, which can realize batch cryopreservation of cell samples by coupling a single liquid filling system.

[0052] It should be noted that a cryogenic pulsating heat pipe (CPHP) is an efficient heat transfer element. Using a cryogenic working fluid such as nitrogen as the working fluid, the pressure difference generated by the continuous phase change of the working fluid at the evaporation end and the condensation end drives the gas-liquid plugs formed by surface tension in the pipe to move. Under a certain heat load, local oscillating flow and overall unidirectional circulation flow can be generated, so as to continuously transfer heat from the evaporation section to the condensation section. In addition to phase change heat transfer, the sensible heat transfer caused by the oscillation of the gas-liquid plugs is also very significant, which makes the pulsating heat pipe have a strong heat transfer ability and can provide an ultra-high heat transfer coefficient between the sample and the working fluid.

[0053] As Figure 3 shown, specifically, grooves are milled on the upper surface of the cell container 22. The size and depth of the grooves are designed according to the amount of cell solution. During operation, the cell solution 24 is dropped into the grooves and then covered by the cover plate 25. In addition, the cell container 22 has 10 completely through circular channels with a diameter of 1 mm, that is, channels. The center spacing of the circular channels is the same as the center spacing of the grooves on the copper plates at the evaporation end and the condensation end. One end of each capillary section is soldered into the groove of the copper plate at the evaporation end 23 or the condensation end 21, and the other end is silver soldered to the circular channel of the cell container 22.

[0054] As Figure 4As shown, in a feasible embodiment of the present utility model, the filling system 1 includes a nitrogen cylinder 11, a gas pipeline 12, a stop valve 13 and a filling pipe 14. The nitrogen cylinder 11 is used to provide nitrogen with a purity of 99.999%. One end of the gas pipeline 12 is connected to the nitrogen cylinder 11 and is used to transport nitrogen from the nitrogen cylinder 11 to other parts of the system. The gas pipeline 12 is a channel for the flow of nitrogen, ensuring that nitrogen can flow smoothly to the place where filling is required. The gas pipeline 12 can use a stainless steel pipe with an outer diameter of 3 mm and an inner diameter of 2 mm. The stop valve 13 is arranged on the gas pipeline 12 and is used to control the flow of nitrogen. By opening or closing the stop valve 13, the filling process of nitrogen can be precisely controlled to ensure that nitrogen is provided when needed and the supply of nitrogen is cut off when not needed. One end of the filling pipe 14 is connected to the other end of the gas pipeline 12, and the other end of the filling pipe 14 is connected to the first end of the first capillary 211. The filling pipe 14 is a direct channel for nitrogen to enter the first capillary 211 from the filling system. Through the filling pipe 14, nitrogen can be precisely transported to the pulsating heat pipe loop.

[0055] It should be noted that since the volume of the filling pipe 14 between the stop valve 13 and the pulsating hot end should be minimized to reduce the error in the calculation of the filling ratio during the experiment, the filling pipe 14 is selected as a stainless steel capillary with an inner diameter of 1 mm and an outer diameter of 1 / 16 inch.

[0056] In a feasible embodiment of the present utility model, it further includes: a multi-channel connector 4, which is used to connect the filling system 1 and the sub-pulsating heat pipe. Each single-layer sub-pulsating heat pipe is connected in parallel through a multi-channel connector 4, and is uniformly connected to the filling system 1 through the filling pipe 14. The advantage of this structure is that for the traditional series structure, each CPHP requires a separate filling system, that is, multiple filling systems are required to cool multiple groups of cell solutions. In the present utility model, each single-layer sub-CPHP is connected in parallel through the connector, and they can share the same filling system, realizing batch processing of samples while simplifying the experimental device.

[0057] The multi-channel connector 4 includes a first connection section 41 and a second connection section 42. The first connection section 41 is connected to the filling pipe 14; the second connection section 42 is horizontally provided with multiple ones, and the second connection section 42 is vertically connected to the first connection section 41; the second connection section 42 is used to communicate with the first end of the first capillary 211. Flow channels 43 are arranged inside both the first connection section 41 and the second connection section 42. The multi-channel connector 4 is made of stainless steel and has a flow channel with a diameter of 1 mm inside. During assembly, the first capillary 211 and the filling pipe 14 are inserted into the grooves (with a diameter of 1.64 mm, slightly larger than the outer diameter of the pipe) at the outlet sections of each channel, and are silver-soldered at the chamfer 44. Finally, in each single-layer PHP, the multi-channel connector 4, the capillary section, and the cell container 22 together form a continuous annular flow channel.

[0058] As Figure 4 shown, in a feasible embodiment of the present utility model, the filling system 1 further includes a buffer tank 15, a molecular pump unit 16, a first valve 17, a second valve 18, a first pressure sensor 19 and a second pressure sensor 110. The buffer tank 15 is arranged on the gas pipeline 12; the molecular pump unit 16 is connected in parallel with the nitrogen cylinder 11; the first valve 17 is arranged between the molecular pump unit 16 and the gas pipeline 12; the second valve 18 is arranged between the nitrogen cylinder 11 and the gas pipeline 12. The first pressure sensor 19 is arranged at one end of the gas pipeline 12 close to the filling pipe 14 for detecting the pressure fluctuation at the condensation end; the second pressure sensor 110 is communicated with the buffer tank 15 for detecting the pressure of the buffer tank 15 to adjust the liquid filling rate.

[0059] Referring again to Figure 3 shown, in a feasible embodiment of the present utility model, the second capillary tubes 231 are all in a U shape, the first capillary tube 211 includes a U-shaped tube 2111 and an L-shaped tube 2112, and there are two L-shaped tubes 2112, which are respectively located on both sides of the condensation end 21, and there are multiple U-shaped tubes 2111, and the multiple U-shaped tubes 2111 are all located between the two L-shaped tubes 2112.

[0060] In a feasible embodiment of the present utility model, the condensation end 21, the cell container 22 and the evaporation end 23 are all made of oxygen-free copper plates, which have a high thermal conductivity and improve the heat transfer efficiency.

[0061] In a feasible embodiment of the present utility model, a heating film 232 is arranged on the back of the evaporation end 23 to provide a thermal driving force for the working medium in the CPHP during the experiment, so that the gas-liquid plugs inside form a circulating flow. The heating film 232 can adopt a polyimide heating film.

[0062] Referring again to Figure 1 shown, in a feasible embodiment of the present utility model, the outer peripheries of the cell container 22 and the evaporation end 23 are wrapped with a foam layer 32 for heat insulation and anti-pollution; a support structure 33 is arranged at the bottoms of the cell container 22 and the evaporation end 23 to make the structure of the cell container 22 firm.

[0063] It should be noted that the pre-cooling system 3 may include a Dewar 34, and liquid nitrogen is introduced into the Dewar 34 through a liquid inlet pipe 35.

[0064] As Figures 1 - 5 shown, the working process of the cell vitrification cryopreservation device provided by the present utility model is as follows:

[0065] Step 1: Transfer the cell solution 24 to the cell container 22 of the pulsating heat pipe system 2, cover the cover plate 25, and wrap the evaporation end 23 and the cell container 22 parts of the whole pulsating heat pipe system 2 with the foam layer 32.

[0066] Step 2: Open the first valve 17 and the stop valve 13 in the filling system 1, close the second valve 18, turn on the molecular pump unit 16, and evacuate the atmosphere in the gas pipeline 12 and the internal flow channels of the pulsating heat pipe system 2.

[0067] Step 3: After the gas pipeline 12 and the pulsating heat pipe system 2 have a good vacuum degree (<10-1 Pa), turn off the molecular pump unit 16 and the first valve 17, open the nitrogen cylinder valve, the stop valve 13 and the second valve 18, and introduce high-purity nitrogen to flush the pipeline and the pulsating heat pipe loop.

[0068] Step 4: After flushing with nitrogen for more than ten seconds, close the nitrogen cylinder valve and the second valve 18, and open the first valve 17, the stop valve 13 and the molecular pump unit 16 again to evacuate the nitrogen inside the pipeline.

[0069] Step 5: Repeat Step 3 and Step 4 five times. After the last time using the molecular pump unit 16 to evacuate the gas pipeline 12 and the pulsating heat pipe system 2 to a high vacuum (<10-5 Pa), close all valves;

[0070] The above Steps 1 - 5 are carried out using the filling system 1, aiming to purge and purify the pipeline to ensure that there are no impurities in the pipeline during the cooling process and avoid pipeline blockage.

[0071] Step 6: Introduce liquid nitrogen from the liquid inlet pipe 35 into the Dewar 34.

[0072] Step 7: Start the heater to provide a heat input of 1 W to the evaporation end 23 of each sub-pulsating heat pipe.

[0073] Step 8: Open the stop valve 13 and the second valve 18 in the filling system 1, and the nitrogen cylinder valve, introduce high-purity nitrogen into the pulsating heat pipe system 2. After the internal pressure P2 of the buffer tank 15 is in the pressure range corresponding to a liquid filling rate of 30% - 50% (experiments show that the pulsating heat pipe has the best heat transfer performance in this liquid filling rate range), close the nitrogen cylinder valve, the stop valve 13 and the second valve 18.

[0074] Step 9: The nitrogen is quickly pre-cooled and liquefied at the condensation end 21 to form alternately distributed gas-liquid plugs, and driven by the heat input at the evaporation end 23, it quickly flows in the entire pulsating heat pipe loop. When the high-speed moving gas-liquid plugs flow through the internal flow channel of the cell container 22, they exchange heat violently with the cell solution on the upper surface of the cell container 22, realizing the vitrification cryopreservation of the cell solution 24. After the cell solution 24 is cooled, transfer it to liquid nitrogen for storage.

[0075] Therefore, for the cryopreservation device for cells provided by the present utility model, the tubular pulsating heat pipe is coupled for the cryopreservation of cells. On the one hand, the problem of limited cooling rate in traditional wet cooling is solved; on the other hand, the advantages of the pulsating heat pipe itself, such as simple structure, low manufacturing cost, and high heat transfer rate, are utilized, and high-rate cooling of cells can be achieved by using a simple operating system.

[0076] In addition, aiming at the problem that the traditional pulsating heat pipe structure cannot process cell samples in batches, an optimized series-parallel structure pulsating heat pipe is provided. Through the multi-channel connector 4, the series structures of several sub-pulsating heat pipes are coupled, enabling them to share the same filling system and pre-cooling system, simplifying the operating system and reducing costs while achieving the cryopreservation of cell samples.

[0077] In the description of the embodiments of the present utility model, it should be noted that unless otherwise clearly specified and defined, the terms "connected" and "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium. For those of ordinary skill in the art, the specific meanings of the above terms in the embodiments of the present utility model can be understood according to specific circumstances.

[0078] In the description of this specification, the descriptions with reference to terms such as "one embodiment", "some embodiments", "mode", "specific mode", or "some modes" etc. mean that the specific features, structures, materials, or characteristics described in connection with the embodiment or mode are included in at least one embodiment or mode of the embodiments of the present utility model. In this specification, the schematic descriptions of the above terms do not necessarily refer to the same embodiment or mode. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or modes. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or modes described in this specification and the features of different embodiments or modes.

[0079] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present utility model and are not intended to limit them; although the present utility model has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present utility model.

Claims

1. A cell vitrification freezing device, characterized in that: include: A charging system (1) for providing nitrogen; A pulsating heat pipe system (2) comprises a plurality of sub-pulsating heat pipes, wherein each layer of the sub-pulsating heat pipes comprises: a condensing end (21), a cell container (22) and an evaporating end (23); a first capillary (211) is provided in the condensing end (21); the first capillary (211) is connected to the filling system (1); a groove is provided in the cell container (22), and a second capillary (231) is provided in the evaporating end (23); the first capillary (211), the groove and the second capillary (231) are connected in sequence to form a pulsating heat pipe loop; a cell solution (24) is contained in the cell container (22), and the cell solution (24) is separated from the groove; The precooling system (3) has a built-in liquid nitrogen pool (31); the condensation end (21) is in contact with the liquid nitrogen pool (31), and the cell container (22) and the evaporation end (23) are both thermally insulated from the liquid nitrogen pool (31).

2. The cell vitrification freezing device according to claim 1, characterized in that: The filling system (1) comprises: A nitrogen cylinder (11) for providing nitrogen with a purity of 99.999%; A gas pipeline (12), one end of which is connected to the nitrogen bottle (11); A stop valve (13) is arranged on the gas pipeline (12); A filling tube (14) has one end in communication with the other end of the gas pipeline (12), and the other end of the filling tube (14) is in communication with the first capillary tube (211).

3. The cell vitrification freezing device according to claim 2, characterized in that: Also includes: A multi-channel connector (4) is used to connect the filling system (1) and the sub-pulsating heat pipe, and the multi-channel connector (4) comprises: A first connecting section (41) connected to the filling pipe (14); A plurality of second connecting sections (42) are arranged horizontally, and the second connecting section (42) is vertically connected to the first connecting section (41); the second connecting section (42) is used to communicate with the first capillary (211).

4. The cell vitrification freezing device according to claim 3, characterized in that: The filling system (1) further comprises: A buffer tank (15) is arranged on the gas pipeline (12); A molecular pump unit (16) connected in parallel with the nitrogen bottle (11); A first valve (17) is arranged between the molecular pump unit (16) and the gas pipeline (12); A second valve (18) is arranged between the nitrogen bottle (11) and the gas pipeline (12).

5. The cell vitrification freezing device according to claim 4, characterized in that: The filling system (1) further comprises: A first pressure sensor (19), arranged at one end of the gas pipeline (12) close to the filling pipe (14), and used to detect pressure fluctuations at the condensation end; A second pressure sensor (110) is in communication with the buffer tank (15) and is used to detect the pressure of the buffer tank (15) to adjust the liquid filling rate.

6. The cell vitrification freezing device according to claim 1, characterized in that: The second capillary tubes (231) are all U-shaped, the first capillary tubes (211) include a U-shaped tube (2111) and an L-shaped tube (2112), there are two L-shaped tubes (2112), and they are respectively located on both sides of the condensation end (21), there are multiple U-shaped tubes (2111), and the multiple U-shaped tubes (2111) are all located between two L-shaped tubes (2112), and the L-shaped tubes (2112) are connected to the filling system (1).

7. The cell vitrification freezing device according to claim 1, characterized in that: The condensation end (21), the cell container (22) and the evaporation end (23) are all made of oxygen-free copper plates.

8. The cell vitrification freezing device according to claim 1, characterized in that: A heating film (232) is provided on the back side of the evaporation end (23).

9. The cell vitrification freezing device according to claim 1, characterized in that: The peripheries of the cell container (22) and the evaporation end (23) are wrapped with a foam layer (32) to provide heat insulation and prevent contamination; and the bottoms of the cell container (22) and the evaporation end (23) are provided with a support structure (33).

10. The cell vitrification freezing device according to claim 3, characterized in that: The gas pipeline (12) and the filling pipe (14) both comprise stainless steel pipes.

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