CN112021308B - Cooling and rewarming system for vitrified preservation of biological materials - Google Patents
Cooling and rewarming system for vitrified preservation of biological materials Download PDFInfo
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- CN112021308B CN112021308B CN202011170707.9A CN202011170707A CN112021308B CN 112021308 B CN112021308 B CN 112021308B CN 202011170707 A CN202011170707 A CN 202011170707A CN 112021308 B CN112021308 B CN 112021308B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
- A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
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Abstract
The invention relates to a cooling and rewarming system for vitrification preservation of biological materials, belonging to the technical field of preservation of biological materials. The system comprises a loop consisting of a rewarming and cooling worktable mechanism, an air source tank, a cooling working medium container, a rewarming working medium container, an external liquid storage tank, an external liquid supply tank, an electric air pump and the like; the rewarming and cooling worktable mechanism comprises a jet mechanism, a clamping mechanism, a controller and a worktable shell. When the device works, the air source tank is opened, high-pressure air in the air source tank is introduced into the cooling working medium container or the rewarming working medium container to press working media out, the high-pressure air flows into the jet generator in the rewarming and cooling working table mechanism through the second three-way valve, high-speed jet of the working media impacts on the sample box, and liquid nitrogen jet cooling or warm water jet rewarming operation is symmetrically carried out on the sample box. The invention reduces the problem of poor consistency caused by manual operation and improves the preservation efficiency.
Description
Technical Field
The invention belongs to the technical field of preservation of biological materials, and particularly relates to a cooling or rewarming system for low-temperature vitrification preservation of biological materials such as cells, tissues, organs and the like.
Background
Cryopreservation is the only effective long-term preservation means at present for a plurality of biological materials, and vitrification preservation is an important method, and the method avoids the dehydration process of the traditional slow cryopreservation method by converting a liquid substance into an amorphous state (glassy state) with extremely high viscosity, so that the internal tissues of the biological samples do not generate (or only generate a small amount of) ice crystals during the preservation process, and the cells are less damaged by low temperature. The key point of realizing vitrification preservation is to carry out rapid cooling and rewarming on a biological sample, the existing method mostly adopts a manual operation mode, a thin tube filled with cells is rapidly put into a liquid nitrogen container to be cooled or liquid drops containing the cells are sequentially dripped into a freezing carrier to realize rapid cooling, and a test tube is placed into a warm water bath to be continuously shaken to heat the cells during rewarming. The manual operation method has the problems of poor consistency, high operation difficulty, low storage efficiency and the like, and in addition, related parameters such as the sample temperature, the environmental temperature and the like in the temperature reduction or rewarming process cannot be recorded, and the parameters have very important significance for the quality analysis of the low-temperature stored sample.
Disclosure of Invention
In order to solve the problems of poor consistency, high operation difficulty, low preservation efficiency, incapability of tracing the preservation process and the like of the existing biological material low-temperature preservation method, the invention provides a cooling and rewarming system for the vitrification preservation of biological materials.
A cooling rewarming system for biological material vitrification preservation includes rewarming cooling worktable mechenism, air source jar 1, cooling working medium container 2, rewarming working medium container 3, outer liquid storage jar 4, outer liquid feed jar 5 and electric air pump 20.
The rewarming and cooling worktable mechanism comprises a jet mechanism, a clamping mechanism, a controller and a worktable shell 81.
The jet flow mechanism comprises a pair of jet flow generating mechanisms, namely an upper jet flow generating mechanism and a lower jet flow generating mechanism; the upper jet flow generating mechanism comprises an upper jet flow generator 813 and an upper splint 83, and the lower jet flow generating mechanism comprises a lower jet flow generator 814 and a lower splint 82; the lower jet flow generation mechanism is fixed in the workbench shell 81, the upper jet flow generation mechanism is movably positioned in the shell 81, and a jet orifice of the upper jet flow generator 813 is vertically corresponding to a jet orifice of the lower jet flow generator 814;
the clamping mechanism comprises an electric cylinder 89, the electric cylinder 89 is fixedly arranged outside the machine shell 81 through an electric cylinder fixing plate 88, a piston rod 86 of the electric cylinder 89 extends into the machine shell 81 of the workbench and is fixedly connected with an upper jet flow generating mechanism to drive the upper jet flow generating mechanism to move relative to a lower jet flow generating mechanism;
the inlet of the upper jet generator 813 and the inlet of the lower jet generator 814 are respectively communicated with one port of a second three-way valve 7, and the other two ports of the second three-way valve 7 are respectively communicated with a cooling working medium container 2 and a rewarming working medium container 3;
the cooling working medium container 2 is communicated with an outer liquid storage tank 4 through a pipeline, and a second air pressure sensor 22 is arranged on the outer liquid storage tank 4; a third electromagnetic valve 17 is arranged on a pipeline between the cooling working medium container 2 and the outer liquid storage tank 4; the rewarming working medium container 3 is communicated with an external liquid supply tank 5 through a pipeline, and a fourth electromagnetic valve 18 is arranged on the pipeline between the rewarming working medium container 3 and the external liquid supply tank 4;
a heating element 12, a temperature sensor 13 and a liquid level sensor 14 are arranged in the rewarming working medium container 3; the external liquid supply tank 5 is positioned above the rewarming working medium container 3;
the gas source tank 1 is respectively communicated with a cooling working medium container 2 and a rewarming working medium container 3 through a pipeline and a three-way pipe; a first electromagnetic valve 15 is arranged on a pipeline between the gas source tank 1 and the cooling working medium container 2, and a second electromagnetic valve 16 is arranged on a pipeline between the gas source tank 1 and the rewarming working medium container 3; one side of the air source tank 1 is communicated with an electric air pump 20 through a pipeline and a fifth electromagnetic valve 19;
placing the cooled or reheated sample box on a lower clamping plate 82 of a lower jet flow generation mechanism, starting a controller, and driving an upper clamping plate 83 of an upper jet flow generation mechanism to move through an electric cylinder 89 to realize corresponding matching clamping of the upper clamping plate 83 and the lower clamping plate 82; and opening the first electromagnetic valve 15, introducing high-pressure gas in the gas source tank 1 into the cooling working medium container 2 or the rewarming working medium container 3 to extrude the working medium, and flowing into the jet generator in the rewarming and cooling working table mechanism through the second three-way valve 7, wherein the jet generator enables high-speed jet of the working medium to symmetrically impact on the sample box 815, so that the liquid nitrogen jet cooling or warm water jet rewarming operation of the sample box is realized.
The technical scheme for further limiting is as follows:
the upper jet flow generation mechanism also comprises a transition plate 85, the transition plate 85 is parallel to the upper clamping plate 83, and the transition plate 85 and the upper clamping plate 83 are fixedly connected by more than two connecting rods; the upper jet flow generator 813 is fixedly arranged on the upper splint 83 and is positioned between the transition plate 85 and the upper splint 83; a through hole is arranged in the middle of the upper splint 83, and a jet port of the upper jet generator 813 is correspondingly communicated with the through hole of the upper splint 83; a piston rod 86 of the electric cylinder 89 extends into the machine shell 81 and is fixedly connected with a transition plate 85 through a threaded connecting piece;
the lower jet generator 814 is fixedly arranged on the lower splint 82; a through hole is formed in the middle of the lower splint 82, and a jet port of the lower jet generator 814 is correspondingly communicated with the through hole of the lower splint 82; the lower clamping plate 82 is fixed in the shell 81 through more than two fixing rods;
the upper clamping plate 83 corresponds to the lower clamping plate 82 up and down, and the distance between the upper clamping plate 83 and the lower clamping plate 82 is 10 mm.
The clamping mechanism further comprises an electric cylinder fixing plate 88, a lower fixing plate 87 and an upper fixing plate 812; the electric cylinder 89 is fixedly arranged outside the shell 81 through an electric cylinder fixing plate 88; a lower fixing plate 87 parallel to the electric cylinder fixing plate 88 is fixedly arranged in the casing 81; the upper fixing plate 812 is located outside the axial end of the electric cylinder 89, and is fixedly connected with the machine shell 81 and the lower fixing plate 87 through four bolts; a pressure sensor 811 and a damper spring 810 are provided in this order between the upper fixing plate 812 and the cylinder axial end of the electric cylinder 89.
The electric cylinder 89 is a servo electric cylinder, the maximum stroke of the piston rod is 50mm, and the speed is 0-60 mm/s.
The bottom of the external liquid supply tank 5 is communicated with the top of the rewarming working medium container 3 through a pipeline, and the bottom of the external liquid supply tank 5 is higher than the top of the rewarming working medium container 3.
The jet time of the upper jet generator 813 and the jet time of the lower jet generator 814 are both 2-10s, and the jet pressure between the upper splint 83 and the lower splint 82 is both 10-100N.
The first solenoid valve 15, the second solenoid valve 16, the third solenoid valve 17, the fourth solenoid valve 18 and the fifth solenoid valve 19 are all one-way solenoid valves.
In the cooling and rewarming system, the working medium used for cooling operation is liquid nitrogen; the working medium used for rewarming operation is water, alcohol solution or salt solution.
The beneficial technical effects of the invention are embodied in the following aspects:
1. the system disclosed by the invention realizes the automation of the vitrification preservation process of the biological material, integrates the operation steps of driving of the working medium, clamping of the sample, supplementing of the working medium and the like into the same equipment, can realize the complete vitrification preservation cooling and rewarming process by operating the controller, greatly reduces the operation difficulty, reduces the problem of poor consistency caused by manual operation, and improves the preservation efficiency. And the data such as pressure, temperature and the like in the operation process can be acquired, and the traceability of the low-temperature storage process is realized.
2. The automatic clamping mechanism of the system can improve the pressure control precision during clamping from 10% of that of a traditional cylinder workbench to 1%, avoids damage to a sample caused by overlarge pressure, the automatic liquid supplementing device enables operators not to need to manually add working media, can reduce potential safety hazards existing when the operators contact low-temperature liquid, and realizes control over the volume and temperature of the working media during supplementing of the temperature-regaining working media.
Drawings
FIG. 1 is a schematic diagram of the system of the present invention.
Fig. 2 is a schematic structural diagram of a cooling and rewarming worktable.
Fig. 3 is a schematic view of the working state of the cooling and rewarming worktable mechanism.
Numbers in fig. 1-3: the device comprises an air source 1, a cooling working medium container 2, a rewarming working medium container 3, an outer liquid storage tank 4, an outer liquid supply tank 5, a first three-way valve 6, a second three-way valve 7, a heating element 12, a temperature sensor 13, a liquid level sensor 14, a first electromagnetic valve 15, a second electromagnetic valve 16, a third electromagnetic valve 17, a fourth electromagnetic valve 18, a fifth electromagnetic valve 19, an electric air pump 20, a first air pressure sensor 21 and a second air pressure sensor 22; the sample holder comprises a workbench housing 81, a lower clamping plate 82, an upper clamping plate 83, a locking nut 84, a transition plate 85, a piston rod 86, a lower fixing plate 87, an electric cylinder fixing plate 88, an electric cylinder 89, a buffer spring 810, a pressure sensor 811, an upper fixing plate 812, an upper jet generator 813, a lower jet generator 814 and a sample box 815.
Detailed Description
The invention will now be further described by way of example with reference to the accompanying drawings.
Referring to fig. 1, a cooling and rewarming system for vitrification preservation of biological materials comprises a rewarming and cooling worktable mechanism, an air source tank 1, a cooling working medium container 2, a rewarming working medium container 3, an external liquid storage tank 4, an external liquid supply tank 5 and an electric air pump 20.
Referring to fig. 2, the rewarming and cooling worktable mechanism includes a jet mechanism, a clamping mechanism, a controller and a worktable casing 81.
The jet mechanism comprises a pair of jet generating mechanisms, namely an upper jet generating mechanism and a lower jet generating mechanism.
The upper jet generating mechanism includes an upper jet generator 813, an upper splint 83 and a transition plate 85. The transition plate 85 is parallel to the upper clamping plate 83, and the transition plate 85 and the upper clamping plate 83 are fixedly connected by more than two connecting rods. The upper jet generator 813 is fixedly mounted to the upper clamp plate 83 and is positioned between the transition plate 85 and the upper clamp plate 83. The middle part of the upper splint 83 is provided with a through hole, and the jet orifice of the upper jet generator 813 is correspondingly communicated with the through hole of the upper splint 83. A piston rod 86 of the electric cylinder 89 extends into the machine shell 81 and is fixedly connected with a transition plate 85 through a threaded connecting piece; the upper jet generating mechanism is movably mounted in the workbench housing 81.
The lower jet generating mechanism includes a lower jet generator 814 and a lower splint 82. The lower jet generator 814 is fixedly mounted on the lower splint 82; a through hole is formed in the middle of the lower splint 82, and a jet port of the lower jet generator 814 is correspondingly communicated with the through hole of the lower splint 82; the lower clamp plate 82 is fixed in the table case 81 by two or more fixing bars.
The jet orifice of the upper jet generator 813 corresponds to the jet orifice of the lower jet generator 814 up and down, the upper splint 83 corresponds to the lower splint 82 up and down, and the distance between the upper splint 83 and the lower splint 82 is 10 mm.
The jet time of the upper jet generator 813 and the jet time of the lower jet generator 814 are the same and are both 2-10s, and the jet pressure between the upper splint 83 and the lower splint 82 is both 10-100N.
The clamping mechanism comprises an electric cylinder 89, an electric cylinder fixing plate 88, a lower fixing plate 87 and an upper fixing plate 812. The electric cylinder 89 is a servo electric cylinder, the maximum stroke of the piston rod is 50mm, and the speed is 0-60 mm/s. The electric cylinder 89 is fixedly arranged outside the machine shell 81 through an electric cylinder fixing plate 88, and a lower fixing plate 87 parallel to the electric cylinder fixing plate 88 is fixedly arranged inside the machine shell 81; the upper fixing plate 812 is located outside the cylinder body axial end of the electric cylinder 89, and is fixedly connected with the machine case 81 and the lower fixing plate 87 by four bolts; a pressure sensor 811 and a damper spring 810 are sequentially mounted between the upper fixing plate 812 and the cylinder axial end of the electric cylinder 89.
The inlet of the upper jet generator 813 and the inlet of the lower jet generator 814 are respectively communicated with one port of the second three-way valve 7, and the other two ports of the second three-way valve 7 are respectively communicated with the cooling working medium container 2 and the rewarming working medium container 3.
The cooling working medium container 2 is communicated with an outer liquid storage tank 4 through a pipeline, and a second air pressure sensor 22 is arranged on the outer liquid storage tank 4; a third electromagnetic valve 17 is arranged on a pipeline between the cooling working medium container 2 and the outer liquid storage tank 4; the rewarming working medium container 3 is communicated with an external liquid supply tank 5 through a pipeline, and a fourth electromagnetic valve 18 is arranged on the pipeline between the rewarming working medium container 3 and the external liquid supply tank 4.
A heating element 12, a temperature sensor 13 and a liquid level sensor 14 are arranged in the rewarming working medium container 3. The external liquid supply tank 5 is positioned above the rewarming working medium container 3; the bottom of the external liquid supply tank 5 is communicated with the top of the rewarming working medium container 3 through a pipeline, and the bottom of the external liquid supply tank 5 is higher than the top of the rewarming working medium container 3.
The gas source tank 1 is respectively communicated with a cooling working medium container 2 and a rewarming working medium container 3 through a pipeline and a three-way pipe; a first electromagnetic valve 15 is arranged on the pipeline between the gas source tank 1 and the cooling working medium container 2, and a second electromagnetic valve 16 is arranged on the pipeline between the gas source tank 1 and the rewarming working medium container 3. One side of the air source tank 1 is communicated with an electric air pump 20 through a pipeline and a fifth electromagnetic valve 19.
The first solenoid valve 15, the second solenoid valve 16, the third solenoid valve 17, the fourth solenoid valve 18, and the fifth solenoid valve 19 are all one-way solenoid valves.
The working principle of the invention is illustrated as follows:
when the system is used for rewarming operation, the working medium used by the system is water; when the system is used for cooling operation, the working medium used by the system is liquid nitrogen.
The sample box 815 to be cooled down or rewarming is placed on the lower splint 82 of the lower jet flow generation mechanism, the controller is started, the upper splint 83 of the upper jet flow generation mechanism is driven to move through the electric cylinder 89, and the corresponding matching clamping of the upper splint 83 and the lower splint 82 is realized, as shown in fig. 3. And opening the first electromagnetic valve 15, introducing high-pressure gas in the gas source tank 1 into the cooling working medium container 2 or the rewarming working medium container 3 to extrude the working medium, and flowing into the jet generator in the rewarming and cooling worktable mechanism through the second three-way valve 7, wherein the jet generator enables high-speed jet of the working medium to symmetrically impact on the sample box 815, so that the cooling or warm water jet rewarming operation of the sample box is realized.
When the heating of the rewarming working medium is started, the heating element 12 is electrified to generate heat, the heat productivity is adjusted in real time according to the temperature data of the temperature sensor 13, the temperature of the rewarming working medium reaches a set value, and the temperature of the rewarming working medium is maintained before the rewarming working medium is discharged.
When the liquid level sensor 14 in the rewarming working medium container 3 detects that the volume of the working medium in the container is lower than a set value, the controller opens the fourth electromagnetic valve 18, the rewarming working medium flows out from the external liquid supply tank 5 under the action of gravity and is supplemented into the rewarming working medium container 3, and when the volume of the working medium reaches the set value, the controller closes the fourth electromagnetic valve 18, and the liquid supplementation is finished;
when the liquid level sensor 14 in the rewarming working medium container 3 detects that the volume of the working medium in the container is lower than a set value, the controller opens the fourth electromagnetic valve 18, the rewarming working medium flows out from the external liquid supply tank 5 under the action of gravity and is supplemented into the rewarming working medium container 3, and when the volume of the working medium reaches the set value, the controller closes the fourth electromagnetic valve 18, and the liquid supplementation is finished.
The second air pressure sensor 22 is installed on the outer liquid storage tank 4, the air pressure inside the outer liquid storage tank 4 is gradually increased after liquid replenishing is started, when the second air pressure sensor 22 detects that the air pressure in the outer liquid storage tank 4 is increased to a set value, the third electromagnetic valve 17 is opened by the controller, a cooling working medium is led into the cooling working medium container 2, then the air pressure in the outer liquid storage tank (4) is gradually reduced, and when the air pressure is reduced to the set value, the third electromagnetic valve 17 is closed, and liquid replenishing is finished.
The gas source tank 1 is respectively communicated with a cooling working medium container 2 and a rewarming working medium container 3 through a pipeline and a three-way pipe; a first electromagnetic valve 15 is arranged on a pipeline between the gas source tank 1 and the cooling working medium container 2, and a second electromagnetic valve 16 is arranged on a pipeline between the gas source tank 1 and the rewarming working medium container 3; and opening the first electromagnetic valve 15, introducing high-pressure gas in the gas source tank 1 into the cooling working medium container 2 or the rewarming working medium container 3 to extrude the working medium, and flowing the working medium into the jet generator in the rewarming and cooling worktable mechanism through the second three-way valve 7, so that the high-speed jet of the working medium impacts on the sample box 815, and liquid nitrogen jet cooling or warm water jet rewarming operation is symmetrically carried out on the sample box.
After the gas in the gas source tank 1 is released, the controller opens the fifth electromagnetic valve 19 to start the electric air pump 20 to supplement the gas to the gas source tank 1, and when the first air pressure sensor 21 detects that the air pressure in the gas source tank 1 reaches a set value, the fifth electromagnetic valve 19 is closed, and the electric air pump 20 stops supplying the gas.
The specific working conditions of the invention are illustrated as follows:
1. the specific operation steps of automatic supplement of the cooling working medium are as follows:
1.1 when the liquid nitrogen working medium in the cooling working medium container is consumed, the controller controls the electromagnetic valve 18 to be automatically opened.
1.2 after the switch is turned on, the pressure in the external liquid storage tank is gradually increased, and the working medium is pressed into the cooling working medium container.
1.3 when the pressure sensor detects that the container is full of cooling working medium, the electromagnetic valve 18 is closed, and the liquid supplementing is finished.
2. The specific operation steps of automatic re-warming working medium supplement are as follows:
2.1 when the liquid level sensor detects that the liquid in the rewarming working medium container is less than a set value, the controller controls the electromagnetic valve 17 to be automatically opened.
2.2 the rewarming working medium in the external liquid supply tank 5 flows into the container due to gravity.
2.3 when the volume of the liquid in the container reaches a preset value, the electromagnetic valve 17 is closed, and the liquid supplementing is finished.
3. The specific operation steps of automatic clamping and unloading of the sample are as follows:
3.1 place the sample cartridge into the fluidic generator 8 in the working chamber.
3.2 during clamping, the electric cylinder push rod of the automatic clamping mechanism moves downwards to drive the upper clamping plate and the lower clamping plate to move in opposite directions so as to clamp the sample box.
3.3 when unloading, the electric cylinder push rod moves upwards, and the sample box can be taken out.
4. The specific operation steps of cryopreservation of biological samples are as follows:
4.1 adding a specific cryoprotectant to the cells and bringing the cells into osmotic equilibrium with the protectant solution.
4.2 attaching thermocouples to the surfaces of both sides of the storage bag containing the cells, then placing the storage bag flat in the sample box, and simultaneously placing thermocouples outside the sample box and in the working chamber. All thermocouples are connected with a temperature measuring device through lead wires.
4.3 the sample cartridge is placed into the fluidic generator 8 in the working chamber and clamped.
4.4 close the working chamber door, open the second electromagnetic valve 16 first, then open the first three-way valve 6 and let in the high-pressure gas, at this moment, the cooling working medium is sprayed onto the sample box through the jet generator 8, the cooling rate can reach 104~105And (3) cooling the sample to the glass transition temperature below-130 ℃ quickly at a temperature of less than min.
4.5 when the preset ventilation time is over, the automatic clamping mechanism is quickly lifted within 2s, and at the moment, the sample box is only required to be taken out of the working cavity and put into the liquid nitrogen storage warehouse to avoid the temperature of the sample from rising again, so that the sample can be stored for a long time.
5. The specific operation steps of rewarming the low-temperature biological sample are as follows:
5.1, starting the rewarming operation, starting the heating element 12 in the rewarming working medium container to work, continuously monitoring the liquid temperature by the temperature sensor, and stopping heating when the preset temperature range is 37-40 ℃.
5.2 opening the third solenoid valve 16 to clear the line.
5.3 the sample capsule containing the biological sample is removed from the liquid nitrogen reservoir and placed rapidly in the fluidic generator 8 in the working chamber.
5.4 closing the working cavity door, opening the first three-way valve 6 and introducing high-pressure gas, and at the moment, spraying the rewarming working medium onto the sample box through the pipeline to quickly raise the temperature of the sample to be above 20 ℃.
5.5 taking out the storage bag in the sample box, the cells can be restored to normal temperature, and the method can be used in the fields of scientific research, clinic and the like.
In addition, the present invention can be used for tissue and organ preservation, in addition to cryopreservation of cell suspensions, and the specific procedures are the same as those described above.
Claims (5)
1. The utility model provides a cooling rewarming system for biomaterial vitrification is preserved which characterized in that: comprises a rewarming and cooling worktable mechanism, an air source tank (1), a cooling working medium container (2), a rewarming working medium container (3), an outer liquid storage tank (4), an outer liquid supply tank (5) and an electric air pump (20);
the rewarming and cooling workbench mechanism comprises a jet mechanism, a clamping mechanism, a controller and a workbench shell (81);
the jet flow mechanism comprises a pair of jet flow generating mechanisms, namely an upper jet flow generating mechanism and a lower jet flow generating mechanism;
the upper jet flow generating mechanism comprises an upper jet flow generator (813) and an upper splint (83), and the lower jet flow generating mechanism comprises a lower jet flow generator (814) and a lower splint (82); the lower jet flow generation mechanism is fixed in the workbench shell (81), the upper jet flow generation mechanism is movably positioned in the workbench shell (81), and a jet flow port of the upper jet flow generator (813) is vertically corresponding to a jet flow port of the lower jet flow generator (814);
the upper jet flow generation mechanism also comprises a transition plate (85), the transition plate (85) is parallel to the upper clamping plate (83), and the transition plate (85) and the upper clamping plate (83) are fixedly connected by more than two connecting rods; the upper jet flow generator (813) is fixedly arranged on the upper splint (83) and is positioned between the transition plate (85) and the upper splint (83); a through hole is arranged in the middle of the upper splint (83), and a jet port of the upper jet generator (813) is correspondingly communicated with the through hole of the upper splint (83); a piston rod (86) of the electric cylinder (89) extends into the workbench shell (81) and is fixedly connected with a transition plate (85) through a threaded connecting piece;
the lower jet generator (814) is fixedly arranged on the lower splint (82); a through hole is formed in the middle of the lower splint (82), and a jet port of the lower jet generator (814) is correspondingly communicated with the through hole of the lower splint (82); the lower splint (82) is fixed in the workbench shell (81) through more than two fixing rods;
the upper clamping plate (83) and the lower clamping plate (82) correspond to each other up and down;
the clamping mechanism comprises an electric cylinder (89), the electric cylinder (89) is fixedly arranged outside the workbench casing (81) through an electric cylinder fixing plate (88), a piston rod (86) of the electric cylinder (89) extends into the workbench casing (81) and is fixedly connected with the upper jet flow generating mechanism to drive the upper jet flow generating mechanism to move relative to the lower jet flow generating mechanism;
the clamping mechanism further comprises an electric cylinder fixing plate (88), a lower fixing plate (87) and an upper fixing plate (812); a lower fixing plate (87) which is correspondingly parallel to the electric cylinder fixing plate (88) is fixedly arranged in the workbench shell (81); the upper fixing plate (812) is positioned outside the axial end part of the cylinder body of the electric cylinder (89), and is fixedly connected with the workbench shell (81) and the lower fixing plate (87) through four bolts; a pressure sensor (811) and a buffer spring (810) are sequentially arranged between the upper fixing plate (812) and the axial end part of the cylinder body of the electric cylinder (89);
the inlet of the upper jet flow generator (813) and the inlet of the lower jet flow generator (814) are respectively communicated with one port of a second three-way valve (7), and the other two ports of the second three-way valve (7) are respectively communicated with a cooling working medium container (2) and a rewarming working medium container (3);
the cooling working medium container (2) is communicated with an outer liquid storage tank (4) through a pipeline, and a second air pressure sensor (22) is arranged on the outer liquid storage tank (4); a third electromagnetic valve (17) is arranged on a pipeline between the cooling working medium container (2) and the outer liquid storage tank (4); the rewarming working medium container (3) is communicated with the external liquid supply tank (5) through a pipeline, and a fourth electromagnetic valve (18) is arranged on the pipeline between the rewarming working medium container (3) and the external liquid supply tank (5);
a heating element (12), a temperature sensor (13) and a liquid level sensor (14) are arranged in the rewarming working medium container (3); the external liquid supply tank (5) is positioned above the rewarming working medium container (3);
the bottom of the external liquid supply tank (5) is communicated with the top of the rewarming working medium container (3) through a pipeline, and the bottom of the external liquid supply tank (5) is higher than the top of the rewarming working medium container (3);
the gas source tank (1) is respectively communicated with a cooling working medium container (2) and a rewarming working medium container (3) through a pipeline and a three-way pipe; a first electromagnetic valve (15) is arranged on a pipeline between the gas source tank (1) and the cooling working medium container (2), and a second electromagnetic valve (16) is arranged on a pipeline between the gas source tank (1) and the rewarming working medium container (3); one side of the air source tank (1) is communicated with an electric air pump (20) through a pipeline and a fifth electromagnetic valve (19);
placing the sample box which is cooled or reheated on a lower clamping plate (82) of a lower jet flow generation mechanism, starting a controller, and driving an upper clamping plate (83) of an upper jet flow generation mechanism to move through an electric cylinder (89) to realize corresponding matching clamping of the upper clamping plate (83) and the lower clamping plate (82); opening a first electromagnetic valve (15), introducing high-pressure gas in a gas source tank (1) into a cooling working medium container (2) or a rewarming working medium container (3) to press working media out, and flowing into a jet generator in the rewarming and cooling worktable mechanism through a second three-way valve (7), wherein the jet generator enables high-speed jet of the working media to symmetrically impact a sample box (815), so that liquid nitrogen jet cooling or warm water jet rewarming operation on the sample box is realized;
in the cooling and rewarming system, the working medium used for cooling operation is liquid nitrogen; the working medium used for rewarming operation is water, alcohol solution or salt solution.
2. The cooling and rewarming system for the vitrification preservation of biological materials of claim 1, wherein: the distance between the upper clamping plate (83) and the lower clamping plate (82) is 10 mm.
3. The cooling and rewarming system for the vitrification preservation of biological materials of claim 1, wherein: the electric cylinder (89) is a servo electric cylinder, the maximum stroke of the piston rod is 50mm, and the speed is 0-60 mm/s.
4. The cooling and rewarming system for the vitrification preservation of biological materials of claim 1, wherein: the jet time of the upper jet generator (813) and the jet time of the lower jet generator (814) are both 2-10s, and the jet pressure between the upper splint (83) and the lower splint (82) is both 10-100N.
5. The cooling and rewarming system for the vitrification preservation of biological materials of claim 1, wherein: the first electromagnetic valve (15), the second electromagnetic valve (16), the third electromagnetic valve (17), the fourth electromagnetic valve (18) and the fifth electromagnetic valve (19) are all one-way electromagnetic valves.
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| 玻璃化保存降温过程中微通道的热应力数值模拟;王洋等;《中国科技论文》;20171123(第22期);摘要 * |
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