CN203952247U - Biomaterial low temperature preservation device - Google Patents
Biomaterial low temperature preservation device Download PDFInfo
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- CN203952247U CN203952247U CN201420409821.6U CN201420409821U CN203952247U CN 203952247 U CN203952247 U CN 203952247U CN 201420409821 U CN201420409821 U CN 201420409821U CN 203952247 U CN203952247 U CN 203952247U
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Abstract
The utility model discloses a kind of biomaterial low temperature preservation device.This device comprises multiple solution storage tanks, multi-way switching valve, liquid nitrogen container, peristaltic pump, magnetic valve, waste collection bucket, heat exchanger, industrial control computer, data acquisition and control interface plate, stirs blower fan, electric heater, frozen container, support, incubator, nozzle, temperature sensor, solution conveyor tube, liquid nitrogen carrier pipe etc.Twin-channel peristaltic pump pumps low temperature protectant solution through multi-way switching valve from solution storage tank; the cold that solution is provided by liquid nitrogen in heat exchanger is lowered the temperature; enter subsequently the frozen container in incubator; peristaltic pump is expelled to waste collection bucket by original liquid in container simultaneously; industrial control computer is controlled by many places temperature such as air in collection liquid nitrogen carrier pipe surface, low temperature protectant solution, incubator; realize the Collaborative Control of biomaterial low temperature protectant solution temperature of living in and concentration, reach the object that improves biomaterial low temperature preservation quality.
Description
Technical field
The utility model relates to a kind of low temperature storage device, preserves for the low temperature of the biomaterial that activity after rewarming is had relatively high expectations, routine techniques is difficult to meet the demands.
Background technology
For having the active cell requiring and the preservation of tissue, low temperature may be necessary condition.In order to set up stock, general-80 DEG C of following low temperature refrigerators or the liquid nitrogen of-196 DEG C of using, can provide the almost storage life of endless from available data the latter.Up to the present, people can successfully preserve multiple useful human body cell and thin layer tissue (as blood cell, sperm, ovum, embryo, various Gan Xi Bao ﹑ Gu Sui ﹑ Pi Fu ﹑ Jiao Mo ﹑ blood vessel and cardiac valves etc.), the preservation of these materials has brought great variety to clinical medicine technology, has had influence on entire society.In addition; at agricultural livestock; the Fast-propagation of animals and plants improved seeds; aspect environmental protection, the preservation of animals and plants germ plasm resource in imminent danger and artificial breeding, at food manufacturing and liquor industry; the preservation of strain excellent and continuity; in biomedical sciences research, the maintenance of typical culture, low temperature is preserved application very widely.
In the process of preserving at low temperature; cell can experience from room temperature to low temperature, from low temperature again to the variations in temperature of room temperature; for avoiding cell to come to harm at low temperatures; add a certain amount of cryoprotective agent (molecule as little in glycerine, methyl-sulfoxide etc.; the large molecule such as albumin, HES) be conventional way; before from room temperature to low temperature, load cryoprotective agent, wash-out cryoprotective agent from low temperature returns to room temperature.Cryoprotective agent often has toxic and side effect to cell; the factors such as this toxic action and protectant physicochemical property, temperature, concentration and time have relation; universal law is, temperature is higher, concentration is larger, the cell time is therein longer, and the suffered toxic damages of cell is larger.In temperature during lower than zero degree, inside and outside cell, may freeze, ice crystal can bring a series of impaired consequences of cell that make such as born of the same parents' inner structure destruction, protein denaturation, for the biological tissue that has certain volume, ice crystal also may make tissue occur micro-crack, and intercellular contact is damaged.Vitrification is a kind of by the Cryopreservation of avoiding ice crystal to form with high concentration cryoprotective agent; its key of successfully preserving biomaterial is how to reduce inner to biomaterial and load cryoprotective agent and the toxic damages of cryoprotective agent to cell from the process of biomaterial inside wash-out cryoprotective agent.For the biological tissue that has certain size, cryoprotective agent enter its center Place cell compare enter thin layer tissue or suspension cell institute's time spent much longer, toxic damages problem is even more serious.In order to alleviate toxic damages; can utilize cryoprotective agent toxicity to reduce with concentration and temperature the feature reducing; in the time loading cryoprotective agent to biomaterial is inner, take cooling limit, limit to improve the mode of concentration; finally reach vitrifying desired concn; and take intensification limit, limit to reduce the mode of concentration when from the inner wash-out cryoprotective agent of biomaterial, finally make concentration be close to zero.The automation equipment that the utility model is aforesaid operations is realized provides a kind of technical scheme.
Utility model content
The purpose of this utility model is to provide a kind of biomaterial low temperature preservation device, and this device can make biomaterial under controlled temperature and concentration conditions, carry out loading and the wash-out of cryoprotective agent, preserves its activity thereby farthest realize.
A kind of biomaterial low temperature preservation device, it mainly comprises solution concentration control section and solution temperature control section, and solution concentration control section comprises multiple solution storage tanks, multi-way switching valve, peristaltic pump, frozen container, waste collection bucket, data acquisition and control interface plate, industrial control computer, solution conveyor tube;
Solution temperature control section comprises described frozen container, temperature sensor, liquid nitrogen container, heat exchanger, magnetic valve, stirring blower fan, electric heater, described data acquisition and control interface plate, described industrial control computer, incubator, liquid nitrogen carrier pipe, nozzle; The solution side of solution storage tank, multi-way switching valve, peristaltic pump, heat exchanger, frozen container, waste collection bucket connect with solution conveyor tube in turn; The liquid nitrogen side of liquid nitrogen container, magnetic valve, heat exchanger connects with liquid nitrogen carrier pipe in turn; Industrial control computer is connected with data acquisition and control interface plate, and data acquisition and control interface plate is connected with multi-way switching valve, magnetic valve, electric heater, temperature sensor; Nozzle is positioned at the bottom of incubator, and stirring blower fan is positioned at the top of incubator.
Described solution storage tank is definite according to the low temperature preservation scheme of biomaterial, and quantity is 2-10, and the maximum way of multi-way switching valve is 10.
Described frozen container is placed on support, can take out from support, and support is placed in incubator.
The beneficial effects of the utility model:
The utility model regulates the temperature of solution in frozen container by the measure of two aspects; make low temperature protectant solution be down to design temperature before the frozen container of injection by the heat exchanger of liquid nitrogen cooling on the one hand, on the other hand liquid nitrogen is sprayed into incubator shuttle space temperature is around maintained near design temperature.
In the utility model, in frozen container, the concentration of solution changes in time in gradient; the method that concentration raises is that the low temperature protectant solution of higher concentration is pumped into frozen container; original solution in frozen container is pumped simultaneously; the method that concentration reduces is that the low temperature protectant solution of lower concentration is pumped into frozen container; original solution in frozen container is pumped, new solution all can keep a period of time of setting in advance after entering frozen container simultaneously.
In the utility model, biomaterial is placed in frozen container and lowers the temperature; reach after vitrifying desired concn in biomaterial inner cryogenic protectant solution concentration; generally frozen container should be transferred to low temperature refrigerator or storage liquid nitrogen container; but when condition does not possess, also can increase the amount of liquid nitrogen that sprays into incubator, make the temperature inside the box reach the temperature level of low temperature refrigerator.
Brief description of the drawings
Fig. 1 is a kind of biomaterial low temperature preservation apparatus structure schematic diagram,
In figure: solution storage tank (1-1,1-2,1-5,1-6), multi-way switching valve 2, liquid nitrogen container 3, peristaltic pump 4, magnetic valve (5-1,5-2), waste collection bucket 6, heat exchanger 7, industrial control computer 8, data acquisition and control interface plate 9, stirring blower fan 10, electric heater 11, frozen container 12, support 13, incubator 14, nozzle 15, the first temperature sensor T1, the second temperature sensor T2, three-temperature sensor T3, the 4th temperature sensor T4.
Embodiment
As shown in drawings, a kind of biomaterial low temperature preservation comprises multiple solution storage tank (1-1 with device, 1-2, 1-5, 1-6, only shows 4 here, can as many as 10), the maximum way of multi-way switching valve 2(is 10), liquid nitrogen container 3(is from boosting type or electric boosting type liquid nitrogen container, disposes manometer and safety valve that producer installs on tank) peristaltic pump 4(is binary channels type, in the time using the peristaltic pump tube of specification of the same race, the liquid inventory of these two passages equates), magnetic valve 5-1(is low form, i.e. failure-free operation for a long time under liquid nitrogen temperature), magnetic valve 5-2, waste collection bucket 6, heat exchanger 7, industrial control computer 8, data acquisition and control interface plate 9, incubator 14, electric heater 11, frozen container 12, temperature sensor T1(is the sensor that can measure liquid nitrogen temperature, as RTD, T-shaped thermocouple, K type thermocouple, after data acquisition and control interface plate conversion, its integrated temperature measurement accuracy should reach ± and 0.5 DEG C), temperature sensor T2, temperature sensor T3, temperature sensor T4, liquid nitrogen container 3 tops are provided with safety valve, in incubator 14, be provided with the support of laying frozen container 12, solution storage tank is parallel to be connected with multi-way switching valve 2, one of them passage of the outlet access binary channels peristaltic pump 4 of multi-way switching valve 2, this channel outlet connects the solution side entrance of heat exchanger 7, the solution side outlet of heat exchanger 7 connects frozen container 12, and the solution of frozen container 12 goes out another passage that pipe connects binary channels peristaltic pump 4, and this channel outlet connects waste collection bucket 6, liquid nitrogen container 3 parallel join magnetic valves, one of them magnetic valve 5-1 outlet connects nozzle 15, and liquid nitrogen is sprayed into incubator 14, and another one magnetic valve 5-2 outlet is connected to the liquid nitrogen side of heat exchanger 7, and liquid nitrogen becomes nitrogen and drains into atmosphere after cooling solution, the first temperature sensor T1 is arranged in the solution side outlet of heat exchanger 7, the solution that the second temperature sensor T2 is arranged in frozen container 12 goes out on pipe pipeline, three-temperature sensor T3 is arranged in incubator 14, the 4th temperature sensor T4 is arranged on the liquid nitrogen conveyance conduit being connected with magnetic valve, these temperature sensor signals enter data acquisition and control interface plate 9, data acquisition and control interface plate 9 is connected with industrial control computer 8, and data acquisition and control interface plate 9 is connected with multi-way switching valve 2, magnetic valve, electric heater 11.
The utility model is mainly made up of two parts: solution temperature control and solution concentration Collaborative Control, there is restricting relation in the variation that refers to concentration, temperature, specifically, in the process of cooling, concentration increases gradually, in the process heating up, concentration reduces gradually, in cooling or arbitrary moment of heating up, the temperature of solution is all higher than the freezing temperature of solution.
The effect of solution temperature control section is to make temperature according to setting-up time curve Gradient Descent or rising, comprises described frozen container 12, temperature sensor, liquid nitrogen container 3, heat exchanger 7, magnetic valve 5, stirs blower fan 10, electric heater 11, described data acquisition and control interface plate 9, described industrial control computer 8, incubator 14, liquid nitrogen carrier pipe, nozzle 15.
The effect of solution concentration control section is to make concentration according to setting-up time curve step increase or minimizing, comprises multiple solution storage tanks, multi-way switching valve 2, peristaltic pump 4, frozen container 12, waste collection bucket 6, data acquisition and control interface plate 9, industrial control computer 8, solution conveyor tube.
Solution temperature control section and solution concentration control section share industrial control computer, data acquisition and control interface plate, temperature and concentration over time curve setting by the implement software of calculator.This software is routine techniques to those skilled in the art, so no longer launch.
Carry out the low temperature preservation of biomaterial at application device described in the utility model before; first need according to the kind of the character choose reasonable cryoprotective agent of biomaterial; secondly need to determine that cryoprotective agent loads, elution step according to the Penetration Signature of cryoprotective agent biomaterial; each step comprises the quantitative parameters such as protectant concentration, temperature and duration, and the corresponding relation between low temperature protectant solution chill point temperature and protectant concentration is the basis of carrying out this work.
The course of work of biomaterial being carried out to cryoprotective agent loading is as described below.Biomaterial is placed at the beginning not to be had, in the preservation of cryoprotective agent liquid, to be placed in frozen container, and frozen container is placed on the support in incubator, and solution conveyor tube is connected to frozen container; Under computer control, peristaltic pump is x by concentration
1low concentration low temperature protectant solution storage tank in solution pump, be cooled to a certain temperature T higher than freezing solution point through heat exchanger
s1, deliver to frozen container, solution conveying capacity is slightly more than the required amount of solution of original solution in the frozen container of whole replacements, conveying capacity is suspended peristaltic pump operation after reaching requirement, meanwhile, and under computer control, spray into incubator liquid nitrogen through magnetic valve and nozzle, make the temperature inside the box reach T
s1, biomaterial is T in temperature
s1, concentration is x
1low temperature protectant solution in process setting-up time t
1; Under computer control, peristaltic pump starts again, is x by concentration
2(x
2> x
1) low temperature protectant solution in storage tank, pump, be cooled to a certain temperature T higher than freezing solution point through heat exchanger
s2, deliver to frozen container, solution conveying capacity is slightly more than the required amount of solution of original solution in the frozen container of whole replacements, conveying capacity is suspended peristaltic pump operation after reaching requirement, meanwhile, and under computer control, spray into incubator liquid nitrogen through magnetic valve and nozzle, make the temperature inside the box reach T
s2, biomaterial is T in temperature
s2, concentration is x
2low temperature protectant solution in process setting-up time t
2; Under computer control, repeat above-mentioned steps, improve constantly the concentration of low temperature protectant solution, reduce its temperature, until reach ultimate density x
n, temperature T
sn, biomaterial is processing time t in this solution
n; In incubator, take out frozen container, put it in low temperature refrigerator or liquid nitrogen container and store.Taking the sheep articular cartilage sheet of a certain thickness of modal cryoprotective agent methyl-sulfoxide loading processing as example, said temperature, concentration, processing time step can be as shown in table 1 below:
Table 1
| Step I | Temperature T si(℃) | Concentration x i(%, w/w) | Processing time t i(min) |
| 1 | 22 | 10 | 10 |
| 2 | 22 | 20 | 10 |
| 3 | -5 | 29 | 30 |
| 4 | -8.5 | 38 | 30 |
| 5 | -16 | 47 | 30 |
| 6 | -23 | 56 | 30 |
| 7 | -35 | 63 | 30 |
| 8 | -48.5 | 72 | 30 |
The course of work of biomaterial being carried out to cryoprotective agent wash-out is as described below.Frozen container is placed in low temperature refrigerator or liquid nitrogen container at the beginning, and biomaterial is placed in frozen container, and frozen container is taken out on the support being placed in incubator, and solution conveyor tube is connected to frozen container; Under computer control, peristaltic pump is x by concentration
1high concentration low temperature protectant solution storage tank in solution pump, be cooled to a certain temperature T higher than freezing solution point through heat exchanger
s1, deliver to frozen container, solution conveying capacity is slightly more than the required amount of solution of original solution in the frozen container of whole replacements, conveying capacity is suspended peristaltic pump operation after reaching requirement, meanwhile, and under computer control, spray into incubator liquid nitrogen through magnetic valve and nozzle, make the temperature inside the box reach T
s1, biomaterial is T in temperature
s1, concentration is x
1low temperature protectant solution in process setting-up time t
1; Under computer control, peristaltic pump starts again, is x by concentration
2(x
2< x
1) low temperature protectant solution in storage tank, pump, be cooled to a certain temperature T higher than freezing solution point through heat exchanger
s2deliver to frozen container, solution conveying capacity is slightly more than the required amount of solution of original solution in the frozen container of whole replacements, conveying capacity is suspended peristaltic pump operation after reaching requirement, meanwhile, under computer control, spray into incubator liquid nitrogen through magnetic valve and nozzle, electric heater starts, and under cooling and heating synergy, makes the temperature inside the box reach T
s2, biomaterial is T in temperature
s2, concentration is x
2low temperature protectant solution in process setting-up time t
2; Under computer control, repeat above-mentioned steps, constantly reduce the concentration of low temperature protectant solution, its temperature that raises, until reach ultimate density x
n(being generally 0%), temperature T
sn, biomaterial is processing time t in this solution
n; In incubator, take out frozen container, biomaterial is taken out and does further processing from frozen container.Be treated to example with above-mentioned sheep articular cartilage sheet, its elution process temperature, concentration, processing time step can be
As shown in table 2 below:
Table 2
| Step I | Temperature T si(℃) | Concentration x i(%, w/w) | Processing time t i(min) |
| 1 | -48.5 | 10 | 30 |
| 2 | -35 | 20 | 30 |
| 3 | -23 | 29 | 30 |
| 4 | -16 | 38 | 30 |
| 5 | -8.5 | 47 | 30 |
| 6 | -5 | 56 | 30 |
| 7 | 22 | 63 | 45 |
Claims (4)
1. a biomaterial low temperature preservation device, it is characterized in that: it mainly comprises solution concentration control section and solution temperature control section, solution concentration control section comprises multiple solution storage tanks, multi-way switching valve (2), peristaltic pump (4), frozen container (12), waste collection bucket (6), data acquisition and control interface plate (9), industrial control computer (8), solution conveyor tube; Solution temperature control section comprises described frozen container (12), temperature sensor, liquid nitrogen container (3), heat exchanger (7), magnetic valve (5), stirs blower fan (10), electric heater (11), described data acquisition and control interface plate (9), described industrial control computer (8), incubator (14), liquid nitrogen carrier pipe, nozzle (15);
Solution storage tank, multi-way switching valve (2), peristaltic pump (4), the solution side of heat exchanger (7), frozen container (12), waste collection bucket (6) connect with solution conveyor tube in turn; The liquid nitrogen side of liquid nitrogen container (3), magnetic valve, heat exchanger (7) connects with liquid nitrogen carrier pipe in turn; Industrial control computer (8) is connected with data acquisition and control interface plate (9), and data acquisition and control interface plate (9) is connected with multi-way switching valve (2), magnetic valve, electric heater (11), temperature sensor; Nozzle (15) is positioned at the bottom of incubator (14), and stirring blower fan (10) is positioned at the top of incubator (14).
2. device according to claim 1, is characterized in that: solution storage tank is definite according to the low temperature preservation scheme of biomaterial, and quantity is 2-10, and the maximum way of multi-way switching valve (2) is 10.
3. device according to claim 1, is characterized in that: it is upper that described frozen container (12) is placed on support (13), can take out from support (13), and support (13) is placed in incubator (14).
4. device according to claim 1, it is characterized in that: described device is provided with multiple temperature sensors, wherein the first temperature sensor (T1) is arranged in the solution side outlet of heat exchanger (7), the solution that the second temperature sensor (T2) is arranged in frozen container (12) goes out on pipe pipeline, three-temperature sensor (T3) is arranged in incubator (14), and the 4th temperature sensor (T4) is arranged on the liquid nitrogen conveyance conduit being connected with magnetic valve.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420409821.6U CN203952247U (en) | 2014-07-24 | 2014-07-24 | Biomaterial low temperature preservation device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420409821.6U CN203952247U (en) | 2014-07-24 | 2014-07-24 | Biomaterial low temperature preservation device |
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| CN203952247U true CN203952247U (en) | 2014-11-26 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104986446A (en) * | 2015-05-29 | 2015-10-21 | 浙江省医疗器械研究所 | Automated biological low temperature preservation device |
| CN106190835A (en) * | 2016-07-13 | 2016-12-07 | 天津卫凯生物工程有限公司 | A kind of filling type cell culture system and method |
| CN108441407A (en) * | 2018-04-02 | 2018-08-24 | 天晴干细胞股份有限公司 | Automatic cell purification and cryopreservation device |
| CN109090103A (en) * | 2018-09-05 | 2018-12-28 | 南通普惠精准医疗科技有限公司 | A kind of biological tissue's preservation equipment |
| CN109762730A (en) * | 2018-12-29 | 2019-05-17 | 无锡海核装备科技有限公司 | A kind of biology freezing storage device |
| CN113784618A (en) * | 2019-04-18 | 2021-12-10 | 艾步思国际有限责任公司 | System and process for continuous addition of cryoprotectant |
-
2014
- 2014-07-24 CN CN201420409821.6U patent/CN203952247U/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104986446A (en) * | 2015-05-29 | 2015-10-21 | 浙江省医疗器械研究所 | Automated biological low temperature preservation device |
| CN106190835A (en) * | 2016-07-13 | 2016-12-07 | 天津卫凯生物工程有限公司 | A kind of filling type cell culture system and method |
| CN108441407A (en) * | 2018-04-02 | 2018-08-24 | 天晴干细胞股份有限公司 | Automatic cell purification and cryopreservation device |
| CN109090103A (en) * | 2018-09-05 | 2018-12-28 | 南通普惠精准医疗科技有限公司 | A kind of biological tissue's preservation equipment |
| CN109762730A (en) * | 2018-12-29 | 2019-05-17 | 无锡海核装备科技有限公司 | A kind of biology freezing storage device |
| CN113784618A (en) * | 2019-04-18 | 2021-12-10 | 艾步思国际有限责任公司 | System and process for continuous addition of cryoprotectant |
| CN113784618B (en) * | 2019-04-18 | 2023-12-22 | 艾步思国际有限责任公司 | System and process for continuous addition of cryoprotectant |
| US11889830B2 (en) | 2019-04-18 | 2024-02-06 | Abs Global, Inc. | System and process for continuous addition of cryoprotectant |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141126 Termination date: 20160724 |
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| CF01 | Termination of patent right due to non-payment of annual fee |