CN111357738B - Freezing and thawing control system for human embryo cryopreservation - Google Patents
Freezing and thawing control system for human embryo cryopreservation Download PDFInfo
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- CN111357738B CN111357738B CN202010218701.8A CN202010218701A CN111357738B CN 111357738 B CN111357738 B CN 111357738B CN 202010218701 A CN202010218701 A CN 202010218701A CN 111357738 B CN111357738 B CN 111357738B
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- 230000008014 freezing Effects 0.000 title claims abstract description 123
- 238000007710 freezing Methods 0.000 title claims abstract description 120
- 210000001161 mammalian embryo Anatomy 0.000 title claims abstract description 87
- 238000010257 thawing Methods 0.000 title claims abstract description 45
- 238000005138 cryopreservation Methods 0.000 title claims abstract description 21
- 239000007788 liquid Substances 0.000 claims abstract description 240
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 198
- 238000003860 storage Methods 0.000 claims abstract description 114
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 99
- 239000000523 sample Substances 0.000 claims abstract description 53
- 239000002699 waste material Substances 0.000 claims abstract description 32
- 210000002257 embryonic structure Anatomy 0.000 claims abstract description 25
- 238000004321 preservation Methods 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims description 25
- 238000011010 flushing procedure Methods 0.000 claims description 20
- 230000001681 protective effect Effects 0.000 claims description 20
- 238000002347 injection Methods 0.000 claims description 15
- 239000007924 injection Substances 0.000 claims description 15
- 239000012530 fluid Substances 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- 238000012544 monitoring process Methods 0.000 claims description 3
- 238000005057 refrigeration Methods 0.000 claims description 3
- 238000012546 transfer Methods 0.000 abstract description 8
- 238000005516 engineering process Methods 0.000 description 9
- 238000004017 vitrification Methods 0.000 description 7
- 208000009701 Embryo Loss Diseases 0.000 description 4
- 231100000557 embryo loss Toxicity 0.000 description 4
- 238000012864 cross contamination Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 206010033266 Ovarian Hyperstimulation Syndrome Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000004590 computer program Methods 0.000 description 2
- 239000002577 cryoprotective agent Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000035935 pregnancy Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000258918 Cryptops Species 0.000 description 1
- 238000012425 Freezing-thawing process Methods 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 208000019703 Isolated cryptophthalmia Diseases 0.000 description 1
- 208000034702 Multiple pregnancies Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 239000010808 liquid waste Substances 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
Classifications
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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
-
- 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
- A01N1/0257—Stationary or portable vessels generating cryogenic temperatures
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
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- Thermal Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
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- Environmental Sciences (AREA)
- Mechanical Engineering (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Clinical Laboratory Science (AREA)
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Abstract
The invention discloses a freezing and thawing control system for freezing and thawing human embryos, which comprises a box body, wherein a computer main control system, a display system, a pipeline system and a miniature camera system are arranged in the box body, and the computer main control system controls the whole freezing and thawing process of the embryos; the pipeline system comprises a liquid nitrogen conveying pipe and a plurality of pipelines, one end of the liquid nitrogen conveying pipe is connected with a liquid nitrogen controller, one end of each of the pipelines is connected with a frozen liquid storage bottle, a defreezing liquid storage bottle and a waste liquid storage bottle, and the other ends of the liquid nitrogen conveying pipe and the pipelines are connected with a first needle probe; the inner wall of the box body is provided with a plurality of drawer type units, and a miniature freezing preservation device is arranged in each drawer type unit; the movable track is arranged in the box body, the miniature camera system is arranged on the track, and the miniature camera system moves and rotates along the track at regular time to shoot embryo images in the miniature cryopreservation device, and the shot embryo images are displayed through the display system. The invention can perform integrated operation, does not need multiple transfer, and can freeze multiple embryos at the same time.
Description
Technical Field
The invention belongs to medical equipment, and particularly relates to a control system for vitrification, freezing and thawing of human embryo.
Background
The advent of human embryo freeze thawing technology is a milestone to aid in reproductive development. The first post-freezing embryo transfer pregnancy message was reported by the scholars in australia as early as 1983, after which embryo freeze-thaw techniques developed rapidly. In the treatment cycle of assisted reproduction, redundant embryos often need to be stored, or full embryo storage is needed because fresh period transplantation cannot be carried out due to occurrence of complications after egg taking operation, and embryo freezing and thawing technology is carried out, so that the difficult problem of embryo storage is solved, the occurrence rate of multiple pregnancy rate and ovarian hyperstimulation syndrome (OHSS) is reduced, each oocyte generates higher cumulative Pregnancy Rate (PR), and great economic and social benefits are generated.
At present, embryo cryopreservation mainly has two approaches, namely a programmed freezing technology and a vitrification freezing technology. The programmed freezing technology is to put embryos in cryoprotectants with different concentrations step by step and then put the embryos in a programmed temperature-reducing instrument for slow temperature reduction, gradual dehydration and balance, and the operation is simple, but when the temperature is reduced below the freezing point of the freezing solution, ice crystals can be generated in embryo cells so as to generate physical damage, and meanwhile, the time for waiting for the programmed temperature reduction is too long. The vitrification freezing technology developed in recent decades uses a cryoprotectant with higher concentration to rapidly cool, so that physical damage of ice crystals in the programmed freezing process is effectively avoided, and the vitrification freezing technology becomes an emerging technology for embryo freezing gradually. However, heretofore, vitrification freezing techniques have not had an integrated operating system, the freezing process was done entirely manually, and the technician was required to be high, and the time and effort costs were excessive due to the inability to do so in batch. The two freezing methods need to transfer the embryo for multiple times in the freezing process, so that the risk of embryo loss is increased; the two methods have the problems of freezing and preserving the carrier, namely a closed carrier and an open carrier, the open carrier is rarely used clinically, the closed carrier is mainly a Cryptop carrier, the closed carrier has the problems of long operation time, high technical requirements and high price, and therefore, no carrier is optimal in the aspects of operation, safety, cost and the like.
Consumable materials used in the embryo vitrification freezing process are common culture dishes, freezing liquid is preheated before freezing, then the frozen liquid is sucked into the culture dishes with marks, embryos to be frozen are transferred into different freezing liquids in steps according to required time from a culture system and then transferred into liquid nitrogen, embryos of the same patient can only be frozen once for the safety of samples, even embryos of the same patient can only be frozen for 2-3 embryos at the same time due to the limitation of time, and the patients with multiple embryo freezing often need to be frozen in batches, so the whole freezing process is time-consuming and labor-consuming. When thawing, the embryo needs to be taken out from the liquid nitrogen storage tank, balanced in different gradient thawing solutions and the reverse process of the freezing process is performed, and the problems of time and labor consumption are also caused.
In summary, the existing vitrification freezing technology lacks an integrated operation system, and embryos need to be transferred for multiple times in the freezing process, so that the risk of embryo loss is increased, multiple embryos need to be frozen for multiple times, time and labor are consumed, and operators are easily frostbitten by liquid nitrogen in the operation process.
Disclosure of Invention
The invention aims to: the invention aims to provide a freezing and thawing control system for human embryo cryopreservation, which can perform integrated operation, does not need multiple transfer and can freeze multiple embryos at the same time.
The technical scheme is as follows: the invention comprises a box body, wherein a computer main control system, a display system, a pipeline system, a miniature camera system, a frozen liquid storage bottle, a unfreezing liquid storage bottle and a waste liquid storage bottle are arranged in the box body, and the computer main control system controls the whole freezing and unfreezing process of embryos; the display system displays embryo image information and various monitoring data in the freezing and thawing processes of the embryo;
the pipeline system comprises a liquid nitrogen conveying pipe and a plurality of pipelines, one end of the liquid nitrogen conveying pipe is connected with a liquid nitrogen controller, and the liquid nitrogen controller is externally connected with a liquid nitrogen storage tank; one end of each of the pipelines is connected with a frozen liquid storage bottle, a defreezing liquid storage bottle and a waste liquid storage bottle, and the other end of each of the liquid nitrogen conveying pipes and the other end of each of the pipelines are connected with a corresponding first telescopic needle probe; the liquid nitrogen controller is connected with the computer main control system, and after the balance of the refrigerating fluid is finished, the liquid nitrogen controller gives an instruction to a first needle probe connected with the liquid nitrogen conveying pipe to descend to a target position to start nitrogen injection refrigeration;
a plurality of drawer-type units are arranged on the inner wall of the box body along the circumferential direction, and a miniature freezing preservation device is arranged in each drawer-type unit; a movable track is arranged above the drawer type unit in the box body, the miniature camera system is arranged on the track, and the miniature camera system moves and rotates along the track at regular time to shoot embryo images in the miniature freezing storage device, and the shot embryo images are displayed through a display system; the miniature camera system can observe the state of embryo in the miniature cryopreservation apparatus in real time.
The miniature freezing preservation device comprises a freezing tank, a connecting part and a handle, wherein the connecting part is connected between the freezing tank and the handle; the middle position of the freezing tank is provided with a cylindrical groove, the bottom of the cylindrical groove is provided with a wave roll, the wave roll divides the cylindrical groove into a tank flushing part and an embryo part, the tank flushing part is a liquid suction part of the liquid inlet of the first needle probe, and the embryo part is an embryo placement part; the wave coil limits the embryo to a certain space, and avoids embryo loss in the process of flushing and draining.
The miniature freezing preservation device also comprises a protective cap, the protective cap is sleeved on the freezing tank, and the joint of the protective cap and the freezing tank is closed; a drawer type basket hanging frame is placed in the liquid nitrogen storage tank, a plurality of basket boxes are arranged on the drawer type basket hanging frame, after embryo freezing is finished, a protection cap is sleeved on a freezing pool and then the embryo is stored in the corresponding basket boxes, and a miniature freezing storage device is placed in the liquid nitrogen storage tank through the drawer type basket hanging frame for storage; the protective cap is provided with a low-temperature-resistant position sensor, and the position sensor is used for positioning embryos in the liquid nitrogen storage tank; the protective cap prevents the embryos from cross-contamination with each other in liquid nitrogen.
The computer main control system comprises a power switch, a plurality of valve controllers, a memory and a processor, wherein the valve controllers are respectively arranged on pipelines of a frozen liquid storage bottle, a unfreezing liquid storage bottle and a waste liquid storage bottle and a liquid nitrogen conveying pipe, a time controller and a flowmeter are arranged on the valve controllers, and when a freezing program and a unfreezing program are executed, the time controller uploads an instruction for starting or ending implementation time to the processor in real time, and the processor gives a command for opening and closing the valve to the valve controller; the flowmeter monitors the liquid flow; the whole freezing and thawing process is controlled by the computer main control system, so that the integrated operation is realized.
The liquid nitrogen conveying pipe and the pipeline are provided with a heat preservation layer, a pressure sensor, a temperature sensor and a heating system, wherein the heat preservation layer is used for insulating and preserving heat of the pipeline; the pressure sensor detects pipeline pressure; the temperature sensor detects the temperature of the liquid in the pipeline; the heating system preheats the liquid in the pipeline; the pipeline system transmits the acquired temperature, pressure and flow data to a computer main control system, and the computer main control system gives instructions according to a preset program; realizing automatic operation.
The miniature camera system comprises a microscope and a miniature camera, the focusing distance of the microscope to the embryo is adjusted in real time through the movement and the lifting of the track, so that the embryo image in the miniature freezing storage device is positioned, and the image under the view field of the microscope is shot through the miniature camera; and the state of the embryo in the freezing pool is observed in real time, so that the excessive freezing and the damage to the embryo are avoided.
The nitrogen injection valve is used for completing nitrogen injection according to a program instruction of the computer; the temperature sensor monitors the temperature of the pipeline; the automatic pressure relief valve maintains the pressure of the liquid nitrogen pipeline and the liquid nitrogen container within a safe range; realizing automatic operation.
The second needle type probes are arranged at one ends of the pipelines, which are connected with the frozen liquid storage bottle, the unfreezing liquid storage bottle and the waste liquid storage bottle, and are used for sucking the liquid in the frozen liquid storage bottle and the unfreezing liquid storage bottle; different needle probes can conveniently flush the freezing pool and discharge waste liquid in the limited box space.
The box body is provided with a notch at a position corresponding to the drawer type unit, the miniature freezing preservation device is installed in the drawer type unit through the notch, and a sealing assembly and a spring lock tongue assembly are installed at the contact part of the box body of the drawer type unit and the miniature preservation device; the operation is convenient.
The bottom of the drawer type unit is funnel-shaped, a local net structure is arranged at the position of the bottom of the drawer type unit corresponding to the embryo part of the freezing tank, the bottom of the drawer type unit is connected with the top of the cylinder groove of the freezing tank in a buckling manner, and the joint of the drawer type unit and the freezing tank is closed; the local reticular structure and the wave rolls limit the embryo in a certain space together, so as to avoid embryo loss.
The beneficial effects are that: compared with the prior art, the invention has the beneficial effects that: (1) The automation of the freezing and thawing process is realized rapidly and effectively; (2) The micro freezing preservation device is not only an upper carrier in the freezing thawing process, but also a freezing preservation carrier, so that the risk of embryos in the multiple transferring process is reduced; (3) The use cost of consumable materials is reduced, the miniature freezing preservation device is small in size and flexible to store and take, the space for preserving embryos in a liquid nitrogen tank is saved, and meanwhile, the risks of being frostbitten by liquid nitrogen and hypoxia caused in the process of volatilizing the liquid nitrogen in the operation process of personnel are also reduced; (4) The protective cap plays a good sealing role, cross contamination in liquid nitrogen is avoided, and meanwhile, the protective cap is provided with a positioning device, so that the embryo is positioned conveniently; (5) Novel structure, degree of automation are high, easy operation, factor of safety is high.
Drawings
FIG. 1 is a schematic diagram of the structure of the present invention;
FIG. 2 is a schematic view of a cross section A-A' of the case of the present invention;
FIG. 3 is a schematic view of a micro-preservation device according to the present invention inserted into a drawer unit;
FIG. 4 is a schematic view of the insertion drawer unit of the micro-preservation device of the present invention taken along section B-B';
FIG. 5 is a schematic diagram of a micro-scale preservation device according to the present invention;
FIG. 6 is a schematic view of the micro-preservation device of the present invention in a state of being sleeved with a protective cap;
FIG. 7 is a schematic view of a liquid nitrogen basket for storing a micro-preservation device in accordance with the present invention.
Detailed Description
The invention is described in further detail below in connection with the detailed description and the attached drawings.
As shown in fig. 1, the invention comprises a box body 1, wherein the box body 1 comprises a computer main control system 3, a display system 4, a pipeline system and a miniature camera system 6 which are mutually connected, a frozen liquid storage bottle 10, a unfreezing liquid storage bottle 11 and a waste liquid storage bottle 12, the pipeline system comprises a liquid nitrogen conveying pipe 7 and a plurality of pipelines 13, one end of the liquid nitrogen conveying pipe 7 is connected with a liquid nitrogen controller 8, and the liquid nitrogen controller 8 is externally connected with a liquid nitrogen storage tank; one end of each of the plurality of pipelines 13 is connected with a frozen liquid storage bottle 10, a thawing liquid storage bottle 11 and a waste liquid storage bottle 12, the other end of the liquid nitrogen conveying pipe 7 and the other end of each of the plurality of pipelines 13 are connected with a corresponding first telescopic needle type probe 15, and the first needle type probes 15 are arranged on the box body 1 through brackets; the liquid nitrogen controller 8 is connected with the computer main control system 3, in this embodiment, the number of the frozen liquid storage bottles 10 is two, the gradient of the frozen liquid in the two frozen liquid storage bottles 10 is different, the number of the unfreezing liquid storage bottles 11 is three, the gradient of the unfreezing liquid in the three unfreezing liquid storage bottles 11 is different, the number of the waste liquid storage bottles 12 is one, each liquid storage bottle is provided with an independent pipeline 13, and one end, connected with the frozen liquid storage bottles 10, the unfreezing liquid storage bottles 11 and the waste liquid storage bottles 12, of each pipeline 13 is provided with a second needle probe 14, and the liquid in the frozen liquid storage bottles 10 and the unfreezing liquid storage bottles 11 is sucked through the second needle probe 14.
As shown in fig. 1, the computer master control system 3 controls the whole freezing and thawing process of the embryo; the computer main control system 3 comprises a power switch, a plurality of valve controllers, a memory and a processor, wherein the valve controllers are respectively arranged on pipelines 13 of the frozen liquid storage bottle 10, the unfreezing liquid storage bottle 11 and the waste liquid storage bottle 12, a time controller and a flowmeter are arranged on the valve controllers, when a freezing program and a unfreezing program are executed, the time controller uploads an instruction for starting or ending the implementation time to the processor in real time, and the processor gives the command for the valve controllers to switch the valves; the flow meter monitors the flow of liquid in the pipeline 13 and in the liquid nitrogen delivery pipe 7. The memory stores a computer program and the processor executes the computer program.
The display system 4 displays embryo image information and various monitoring data during freezing and thawing of embryos, and comprises a display and related elements, wherein the display is arranged on the side surface of the shell. The related elements such as a time controller, a temperature sensing device, a pressure sensing device, an image acquisition card, a wireless communicator and the like are respectively arranged at the positions of a refrigerating fluid, a defrosting fluid and a liquid nitrogen conveying pipeline, and all the related elements are connected through wires sleeved with heat insulation layers. The wireless communicator uploads the temperature value, the pressure value, the time and the like to the server, and can record and store relevant data in the embryo freezing process in real time; the image acquisition card can transmit data to the computer main control system 3 and display the data through a display of the display system 4.
As shown in fig. 1, a liquid nitrogen conveying pipe 7 is arranged above a box body 1, one end of the liquid nitrogen conveying pipe 7 is connected with a liquid nitrogen controller 8, a nitrogen injection valve, a temperature sensor and an automatic pressure release valve are arranged on the liquid nitrogen controller 8, and the nitrogen injection valve can complete nitrogen injection according to program instructions of a computer; the temperature sensor detects the temperature of the pipeline; the automatic pressure relief valve maintains the pressure of the liquid nitrogen pipeline and the liquid nitrogen container within a safe range. The liquid nitrogen controller 8 is externally connected with a liquid nitrogen tank, the other end of the liquid nitrogen conveying pipe 7 is connected with a first needle type probe 15 with elasticity, and the other end of the liquid nitrogen conveying pipe 7 is also provided with a valve controller; the liquid nitrogen controller 8 is connected with the computer main control system 3, and after the balance of the refrigerating fluid is finished, the liquid nitrogen controller 8 gives an instruction to the first needle probe 15 connected with the liquid nitrogen conveying pipe 7 to descend to the target position to start nitrogen injection refrigeration; the first needle probe 15 has the same structure as the second needle probe 14, but the first needle probe 15 connected to the liquid nitrogen delivery pipe 7 is resistant to low temperatures.
The liquid nitrogen conveying pipe 7 and the pipeline 13 are respectively provided with an insulating layer, a pressure sensor, a temperature sensor and a heating system, and the insulating layers are used for insulating and preserving heat of the pipeline; the pressure sensor detects the pipeline pressure; the temperature sensor detects the temperature of the liquid in the pipeline; the heating system preheats the liquid in the pipeline; the pipeline system transmits the acquired temperature, pressure and flow data to the computer main control system 3 for recording and storing, and the computer main control system 3 gives instructions according to a preset program.
As shown in fig. 2 to 6, a plurality of drawer units 2 are uniformly arranged on the inner wall of the box body 1 along the circumferential direction, and a micro freezing preservation device 5 is arranged in the drawer units 2; the box body 1 is provided with a notch at a position corresponding to the drawer type unit 2, and the micro freezing preservation device 5 is arranged in the drawer type unit 2 through the notch; the contact part of the drawer type unit box body and the miniature storage device 5 is provided with a sealing component and a spring bolt component. The micro cryopreservation apparatus 5 comprises a freezing tank 21, a connecting part 22, a handle 23 and a protective cap 24, wherein the connecting part 22 is connected between the freezing tank 21 and the handle 23; the middle position of the freezing tank 21 is provided with a cylindrical groove 20, the bottom of the cylindrical groove 20 is provided with a wave roll 19, the wave roll 19 divides the cylindrical groove 20 into a flushing tank part 17 and an embryo part 18, the flushing tank part 17 is a liquid inlet and liquid suction part of the first needle probe 15, and the embryo part 18 is an embryo placement part. The handle 23 can be pasted with embryo information and a two-dimensional code electronic tag with color, and the electronic tag can upload information to a cloud server to generate a human embryo freeze thawing database after scanning. The protective cap 24 is sleeved on the freezing tank 21, and the joint of the protective cap 24 and the freezing tank 21 is closed, so that the risk of cross contamination of embryos in liquid nitrogen can be avoided; as shown in fig. 7, a drawer type basket hanging frame 25 is placed in a liquid nitrogen tank, a plurality of basket boxes 26 are arranged on the drawer type basket hanging frame 25, after embryo freezing is finished, a freezing tank 21 is sleeved with a protective cap 24 and then is stored in the corresponding basket boxes 26, and a micro freezing storage device 5 is placed in the liquid nitrogen tank through the drawer type basket hanging frame 25 for storage, so that a large amount of storage space is saved; the protective cap 24 is provided with a low temperature resistant position sensor which positions embryos in the liquid nitrogen tank.
As shown in fig. 4, the bottom of the drawer unit 2 is funnel-shaped, a local net structure 16 is arranged at the bottom of the drawer unit 2 corresponding to the embryo part 18 of the freezing tank, the bottom of the drawer unit 2 is in snap connection with the top of the cylinder groove 20 of the freezing tank 21, and the joint of the drawer unit 2 and the freezing tank 21 is closed. The partial mesh structure 16 is provided with a water level sensor for sensing the liquid level in the cylinder recess 20.
Since the embryo needs to be always in the liquid environment, the wave coil 19 at the bottom of the cylindrical groove 20 in the freezing tank 21 is of great importance, the wave coil 19 can always ensure the water level of the embryo, and the embryo is locked in a certain space with the local net structure 16 right above the wave coil, so that the embryo is prevented from being lost in the process of flushing and draining. The material of the miniature storage device 5 is low-temperature resistant, the bottom is transparent, the bottom is flat, and a technician can transfer embryos under a microscope.
As shown in fig. 1, a movable rail 9 is arranged above the drawer unit 2 in the case 1, the miniature camera system 6 is arranged on the rail 9, and the miniature camera system 6 moves and rotates along the rail 9 at regular time to shoot embryo images in the miniature cryopreservation apparatus 5, and the shot embryo images are displayed by the display system 4. The micro camera system 6 comprises a microscope and a micro camera, and adjusts the focusing distance of the microscope to the embryo in real time through the movement and the lifting of the track 9, so as to position the embryo image in the micro cryopreservation device 5 and shoot the image under the field of view of the microscope through the micro camera.
When the liquid feeding program is executed, the first needle probe 15 of the target liquid pipeline moves along the track to stretch and retract to the target position, the liquid starts to enter the flushing part 17 of the freezing tank, and when the liquid reaches the sample feeding volume, a signal is transmitted to the controller to close the liquid inlet valve and lift up to the tank body. When the liquid outlet program is executed, the first needle probe 15 on the pipeline 13 connected with the waste liquid storage bottle 12 moves along the track to stretch to a target position, the liquid suction is started to the pool flushing part 17 of the freezing pool, and when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve and lift up to the box body.
The specific operation process is as follows:
1. freezing process
1) Preparing a portable freezer pool: before freezing, firstly, electronic tags with different colors and low temperature resistance are stuck at the handle of the micro freezing preservation device 5, patient information such as names, medical record numbers, embryo periods and the like are marked on the tags, the tags can be printed, codes can be scanned and sent into a server, and a human embryo freezing database is generated. Adding balanced culture solution into embryo part 18 in cylindrical groove 20 of freezing tank 21, checking by two persons, sucking corresponding embryo into embryo part 18 of freezing tank 21 under microscope, transferring embryo-containing freezing tank 21 into incubator, and loading embryo to be frozen together;
2) Freezing and loading machine: firstly, preparing, turning on a power supply, filling a liquid nitrogen storage tank of a refrigerating fluid storage bottle 10 and a liquid waste storage bottle 12, cleaning a pipeline system, inserting a prepared miniature refrigerating storage device 5 into a drawer type unit 2, and preparing for refrigerating; the freezing program is started, the first needle probe 15 on the pipeline 13 of the first frozen liquid storage bottle moves to stretch to the target position, the first needle probe moves to the target position to directly face the flushing part 17 of the freezing tank 21, the valve controller on the first frozen liquid storage bottle is started to start liquid feeding, when the liquid reaches the sample feeding volume, a signal is transmitted to the controller to close the valve controller on the first frozen liquid storage bottle, and the first needle probe 15 corresponding to the first frozen liquid storage bottle is lifted to the box body. The embryo balances in the first refrigerating fluid for a period of time, after the program preset time arrives, the first needle probe 15 on the pipeline 13 of the waste liquid storage bottle moves to stretch to the target position, the flushing part 17 opposite to the refrigerating pool 21 is opened, the liquid outlet valve starts to discharge liquid, the first refrigerating fluid flows into the waste liquid storage bottle 12, when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve, and the first needle probe 15 corresponding to the waste liquid storage bottle 12 is lifted to the box body. The first needle probe 15 on the pipeline 13 of the second frozen liquid storage bottle moves to stretch out and draw back to a target position, the second frozen liquid inlet valve is opened, the second frozen liquid flushing valve transmits a signal to the controller to close the valve controller on the second frozen liquid storage bottle when the liquid reaches the sample injection volume, and the first needle probe 15 corresponding to the second frozen liquid storage bottle is lifted to the box body. When the preset time arrives, the first needle probe 15 on the pipeline 13 of the waste liquid storage bottle moves to stretch to the target position, the flushing part 17 of the freezing tank 21 is opposite, the liquid outlet valve is opened, liquid outlet starts, the second freezing liquid flows into the waste liquid storage bottle 12, when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve, and the first needle probe 15 corresponding to the waste liquid storage bottle is lifted to the box body. Simultaneously, the first needle type probe 15 corresponding to the liquid nitrogen conveying pipe 7 moves and stretches to a target position, the nitrogen injection valve is opened and the liquid nitrogen flushing valve is directly opposite to the flushing tank part 17 of the freezing tank 21, when the nitrogen volume reaches a specified value, a signal can be transmitted to the liquid nitrogen controller to close the nitrogen outlet valve, the first needle type probe 15 corresponding to the liquid nitrogen conveying pipe 7 is lifted to the box body part, and the embryo is frozen; the state of each embryo step during this process can be taken by a microscope and a miniature camera in the box 1 and visualized on the display of the display system 4;
3) Sealing the freezing pool: the handle 23 is held, the micro freezing and preserving device 5 is taken out from the drawer type unit 2, the protecting cap 24 is sleeved, the micro freezing and preserving device 5 is put into a liquid nitrogen transfer box, and all the caps of the micro freezing and preserving device 5 to be frozen together are completed;
4) And (3) preserving liquid nitrogen: the miniature freezing and preserving device 5 covered with the protective cap 24 is put into a liquid nitrogen basket box 26 and put into a liquid nitrogen storage tank for preservation.
2. Thawing process
1) Preparation: before thawing, firstly searching the cryopreservation position and color of a patient to be thawed according to the operation notification sheet and the freezing information, rapidly positioning according to an electronic positioning system provided by a protective cap 24, taking out the patient from a liquid nitrogen storage tank, checking the patient by two persons, putting the patient into a liquid nitrogen transfer box, taking out all thawed embryos in the same batch, putting the embryo into the liquid nitrogen transfer box, and preparing to defrost;
2) Thawing and loading: the power supply is turned on, the unfreezing liquid storage bottle 11 and the waste liquid storage bottle 12 are well filled, the pipeline system is cleaned, a liquid preheating program in the unfreezing process is firstly started before the sample is put on the machine, and the unfreezing liquid storage bottle and the corresponding pipeline are preheated; inserting the prepared micro freezing and preserving device 5 into the drawer type unit 2 to prepare thawing; the first needle probe 15 on the pipeline 13 of the first defrosting liquid storage bottle moves to stretch out and draw back to a target position, the first defrosting liquid inlet valve is opened and transmits a signal to the controller to close the first defrosting liquid inlet valve when the liquid reaches the sample injection volume, and the first needle probe 15 corresponding to the first defrosting liquid storage bottle is lifted to the box body. The embryo balances in the first defrosting liquid for a period of time, after the program preset time is reached, the first needle probe 15 on the pipeline 13 of the waste liquid storage bottle 12 moves to stretch to the target position, the liquid outlet valve is opened to start liquid outlet, the first defrosting liquid flows into the waste liquid storage bottle 12, when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve, and the first needle probe 15 corresponding to the waste liquid storage bottle is lifted to the box body. The first needle probe 15 on the pipeline 13 of the second defrosting liquid storage bottle moves to stretch out and draw back to the target position, the second defrosting liquid inlet valve is opened, the second defrosting liquid is flushed, when the liquid reaches the sample injection volume, a signal is transmitted to the controller to close the second defrosting liquid inlet valve, and the first needle probe 15 corresponding to the second defrosting liquid storage bottle is lifted to the box body. When the preset time arrives, the first needle probe 15 on the pipeline 13 of the waste liquid storage bottle moves to stretch to the target position, the flushing part 17 of the freezing tank 21 is opposite, the liquid outlet valve is opened, liquid begins to be discharged, the second unfreezing liquid flows into the waste liquid storage bottle 12, when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve, and the first needle probe 15 corresponding to the waste liquid storage bottle is lifted to the box body. The first needle probe 15 on the pipeline 13 of the third defrosting liquid storage bottle moves to stretch out and draw back to a target position, the flushing tank part 17 of the freezing tank 21 is opposite, the third defrosting liquid inlet valve is opened, the third defrosting liquid flushing tank transmits a signal to the controller to close the third defrosting liquid inlet valve when the liquid reaches the sample injection volume, and the first needle probe 15 corresponding to the third defrosting liquid storage bottle is lifted to the tank body. When the preset time arrives, the first needle probe 15 on the pipeline 13 of the waste liquid storage bottle moves to stretch to the target position, the flushing part 17 of the freezing tank 21 is opposite, the liquid outlet valve is opened, liquid begins to be discharged, the third unfreezing liquid flows into the waste liquid storage bottle 12, when the liquid outlet volume reaches a specified value, a signal can be transmitted to the controller to close the liquid outlet valve, and the first needle probe 15 corresponding to the waste liquid storage bottle is lifted to the box body. After the thawing, the micro-cryopreservation apparatus 5 is pulled out of the drawer-type unit 2 and put into an incubator for use.
Claims (9)
1. The freezing and thawing control system for human embryo cryopreservation comprises a box body (1), and is characterized in that: the box body (1) is internally provided with a computer main control system (3), a display system (4), a pipeline system, a miniature camera system (6), a frozen liquid storage bottle (10), a unfreezing liquid storage bottle (11) and a waste liquid storage bottle (12), wherein the computer main control system (3) controls the whole freezing and unfreezing process of embryos; the display system (4) displays embryo image information and various monitoring data in the freezing and thawing processes of the embryo;
the pipeline system comprises a liquid nitrogen conveying pipe (7) and a plurality of pipelines (13), one end of the liquid nitrogen conveying pipe (7) is connected with a liquid nitrogen controller (8), and the liquid nitrogen controller (8) is externally connected with a liquid nitrogen storage tank; one end of each of the pipelines (13) is connected with a frozen liquid storage bottle (10), a thawing liquid storage bottle (11) and a waste liquid storage bottle (12), and the other end of the liquid nitrogen conveying pipe (7) and the other end of each of the pipelines (13) are connected with a corresponding first needle type probe (15) with elasticity; the liquid nitrogen controller (8) is connected with the computer main control system (3), and after the balance of the refrigerating fluid is finished, the liquid nitrogen controller (8) gives an instruction to a first needle probe (15) connected with the liquid nitrogen conveying pipe (7) to descend to a target position to start nitrogen injection refrigeration;
a plurality of drawer units (2) are arranged on the inner wall of the box body (1) along the circumferential direction, and a miniature freezing preservation device (5) is arranged in each drawer unit (2); a movable track (9) is arranged above the drawer type unit (2) in the box body (1), the miniature camera system (6) is arranged on the track (9), and the miniature camera system (6) moves and rotates along the track (9) at regular time to shoot embryo images in the miniature freezing storage device (5), and the shot embryo images are displayed through the display system (4);
the miniature cryopreservation device (5) comprises a freezing tank (21), a connecting part (22) and a handle (23), wherein the connecting part (22) is connected between the freezing tank (21) and the handle (23); the middle position of the freezing tank (21) is provided with a cylindrical groove (20), the bottom of the cylindrical groove (20) is provided with a wave roll (19), the wave roll (19) divides the cylindrical groove (20) into a flushing tank part (17) and an embryo part (18), the flushing tank part (17) is a liquid-feeding and liquid-sucking part of the first needle probe (15), and the embryo part (18) is an embryo placement part.
2. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the miniature freezing preservation device (5) further comprises a protective cap (24), the protective cap (24) is sleeved on the freezing tank (21), and the joint of the protective cap (24) and the freezing tank (21) is closed; a drawer type basket hanging frame (25) is placed in the liquid nitrogen storage tank, a plurality of basket boxes (26) are arranged on the drawer type basket hanging frame (25), after embryo freezing is finished, a freezing pool (21) is sleeved with a protective cap (24) and then is stored in the corresponding basket boxes (26), and a miniature freezing storage device (5) is placed in the liquid nitrogen storage tank through the drawer type basket hanging frame (25) for storage; the protective cap (24) is provided with a low-temperature-resistant position sensor, and the position sensor is used for positioning embryos in the liquid nitrogen tank.
3. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the computer main control system (3) comprises a power switch, a plurality of valve controllers, a memory and a processor, wherein the valve controllers are respectively arranged on a frozen liquid storage bottle (10), a unfreezing liquid storage bottle (11), a pipeline (13) of a waste liquid storage bottle (12) and a liquid nitrogen conveying pipe (7), a time controller and a flowmeter are arranged on the valve controllers, when a freezing program and a unfreezing program are executed, the time controller uploads an instruction for starting or ending the implementation time to the processor in real time, and the processor gives a command for the valve controller to switch the valve; the flow meter monitors the flow of liquid.
4. A freeze thaw control system for human embryo cryopreservation according to claim 3 wherein: the liquid nitrogen conveying pipe (7) and the pipeline (13) are respectively provided with a heat preservation layer, a pressure sensor, a temperature sensor and a heating system, and the heat preservation layers are used for insulating and preserving heat of the pipeline; the pressure sensor detects pipeline pressure; the temperature sensor detects the temperature of the liquid in the pipeline; the heating system preheats the liquid in the pipeline; the pipeline system transmits the acquired temperature, pressure and flow data to the computer main control system (3), and the computer main control system (3) gives instructions according to a preset program.
5. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the miniature camera system (6) comprises a microscope and a miniature camera, the distance of focusing of the microscope on the embryo is adjusted in real time through movement and lifting of the track (9), so that the embryo image in the miniature freezing storage device (5) is positioned, and the image under the field of view of the microscope is shot through the miniature camera.
6. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the nitrogen injection valve is arranged on the liquid nitrogen controller (8) and is used for completing nitrogen injection according to program instructions of a computer; the temperature sensor monitors the temperature of the pipeline; the automatic pressure relief valve maintains the pressure of the liquid nitrogen pipeline and the liquid nitrogen container within a safe range.
7. A freeze thaw control system for human embryo cryopreservation according to claim 3 wherein: the one end that a plurality of pipelines (13) are connected with frozen solution storage bottle (10), thawing solution storage bottle (11), waste liquid storage bottle (12) all is equipped with second needle probe (14), absorbs the liquid in frozen solution storage bottle (10), thawing solution storage bottle (11) through second needle probe (14).
8. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the box body (1) is provided with a notch at a position corresponding to the drawer type unit (2), and the miniature freezing preservation device (5) is arranged in the drawer type unit (2) through the notch; the contact part of the drawer type unit box body and the miniature storage device (5) is provided with a sealing component and a spring bolt component.
9. The freeze-thaw control system for human embryo cryopreservation of claim 1 wherein: the bottom of the drawer type unit (2) is funnel-shaped, a local net structure (16) is arranged at the position, corresponding to the embryo part (18) of the freezing tank, of the bottom of the drawer type unit (2), the bottom of the drawer type unit (2) is connected with the top of a cylindrical groove (20) of the freezing tank (21) in a buckling mode, and the joint of the drawer type unit (2) and the freezing tank (21) is airtight.
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| CN106998675A (en) * | 2014-07-30 | 2017-08-01 | 多伦多大学董事局 | System and method for the automatic glass of biomaterial |
| CN212279636U (en) * | 2020-03-25 | 2021-01-05 | 南京鼓楼医院 | Freezing and thawing control system for human embryo freezing |
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| CN215936087U (en) * | 2021-08-30 | 2022-03-04 | 江西楷晶生物科技有限公司 | Freezing and thawing control system for human embryo freezing |
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| US20110207112A1 (en) * | 2008-07-23 | 2011-08-25 | Mariposa Biotechnology, Inc | Automated system for cryopreservation of oocytes, embryos, or blastocysts |
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|---|---|---|---|---|
| CN106998675A (en) * | 2014-07-30 | 2017-08-01 | 多伦多大学董事局 | System and method for the automatic glass of biomaterial |
| CN212279636U (en) * | 2020-03-25 | 2021-01-05 | 南京鼓楼医院 | Freezing and thawing control system for human embryo freezing |
| CN213486620U (en) * | 2020-10-30 | 2021-06-22 | 柳州市工人医院 | Embryo freezing and thawing apparatus |
| CN215936087U (en) * | 2021-08-30 | 2022-03-04 | 江西楷晶生物科技有限公司 | Freezing and thawing control system for human embryo freezing |
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