CN113632785B - Freezing carrying rod and operation method thereof - Google Patents
Freezing carrying rod and operation method thereof Download PDFInfo
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- CN113632785B CN113632785B CN202110943874.0A CN202110943874A CN113632785B CN 113632785 B CN113632785 B CN 113632785B CN 202110943874 A CN202110943874 A CN 202110943874A CN 113632785 B CN113632785 B CN 113632785B
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- 238000007710 freezing Methods 0.000 title claims abstract description 57
- 230000008014 freezing Effects 0.000 title claims abstract description 57
- 238000000034 method Methods 0.000 title claims abstract description 32
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 246
- 239000007788 liquid Substances 0.000 claims abstract description 109
- 238000012546 transfer Methods 0.000 claims abstract description 32
- 239000002577 cryoprotective agent Substances 0.000 claims abstract description 27
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 10
- 238000010257 thawing Methods 0.000 claims description 55
- 230000007704 transition Effects 0.000 claims description 3
- 210000002257 embryonic structure Anatomy 0.000 abstract description 29
- 238000013461 design Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 230000002349 favourable effect Effects 0.000 description 3
- 238000007654 immersion Methods 0.000 description 3
- 208000009701 Embryo Loss Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 231100000557 embryo loss Toxicity 0.000 description 2
- 238000004017 vitrification Methods 0.000 description 2
- 206010042573 Superovulation Diseases 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 238000009499 grossing Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000000638 stimulation Effects 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
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
- A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a frozen carrying rod, which comprises an embryo loading area, a frozen dripping area and a frozen embryo transferring area, wherein two sides of the embryo loading area are respectively communicated with the frozen dripping area and the frozen embryo transferring area, and the embryo loading area is at the lowest position; the method of operation includes transferring embryos with a culture solution into an embryo loading area; removing the excess culture solution; adding the balance liquid drop into the frozen liquid drop zone; allowing the embryo to stand in the balancing solution for T1 min; removing the balance liquid; dropping a cryoprotectant into the frozen drip zone; allowing the embryo to stand in cryoprotectant for T2 min; removing the cryoprotectant; the frozen carrier bar was immersed in liquid nitrogen. When the freezing carrier rod is used for operation, only one transfer operation is needed to be carried out on the embryo, compared with the freezing carrier rod used in the existing freezing operation, the embryo transfer times are reduced, and the damage to the embryo possibly caused by transfer is reduced. Moreover, since only one transfer operation is required for the embryo, the automated design of the operation is facilitated.
Description
Technical Field
The invention relates to the technical field of embryo culture, in particular to a freezing carrier rod and an operation method thereof.
Background
In the course of assisted reproduction for infertility patients, the number of mature ova is generally increased by stimulation of ovulation-promoting drugs, so that ova with a large number of ovaries can be grown at one time. In practice, however, only 1-3 embryos are transferred back to the uterine cavity of a human body in one treatment session. Since implanting too many embryos may increase the risk of multiple fetuses, the remaining embryos may be cryopreserved. Embryo cryopreservation is a well-established method of preserving fertility. The technology is to use a balancing solution and a cryoprotectant to carry out pre-freezing treatment on the embryo according to the operation steps, so that the cryoprotectant gradually permeates into cells of the embryo, the embryo can bear the process of rapidly reducing the temperature without damage, after a series of operation procedures, the embryo can reach a proper frozen state, and the embryo can be stored in a liquid nitrogen environment at the temperature of minus 196 ℃ for a long time. If the period of treatment is unsuccessful, the preserved embryo can be transplanted back to the mother body after the later resuscitation, the superovulation treatment process is not needed, so that a lot of cost can be saved, and the chance of conception of the patient is increased.
At present, in the auxiliary reproduction field, the conventional embryo vitrification freezing method is generally used in the embryo culture process, and the operation flow is generally as follows:
s1, preparing a balance liquid and a culture dish;
s2, preparing balanced liquid drops in a culture dish;
s3, transferring the embryo into an equilibrium droplet by using a Pasteur tube;
s4, allowing the embryo to stand in the balance liquid for 5 minutes;
s5, preparing a cryoprotectant drop in a culture dish;
s6, transferring the embryo into a cryoprotectant drop by using a Pasteur tube;
s7, allowing the embryo to stand in a cryoprotectant for 1 minute;
s8, transferring the embryo to the front end of a freezing carrier rod by using a Pasteur tube;
s9, the front end of the freezing carrier rod is quickly immersed into liquid nitrogen.
In the actual operation process, the embryo vitrification freezing operation flow has the following problems:
because the Pasteur tube is required to transfer the embryo for multiple times in the operation process, the operation steps are complex, and a certain degree of damage or embryo loss can be caused to the embryo, thereby affecting the culture of the embryo and the treatment effect of assisted reproduction. In addition, two consumables, namely a culture dish and a freezing carrier rod, are needed to finish a freezing process.
Disclosure of Invention
The present invention is directed to a freezing carrier bar and an operating method thereof, which solve the above-mentioned problems in the prior art.
In order to achieve the above purpose, the present invention provides the following technical solutions: the frozen carrying rod comprises an embryo loading area, a frozen dripping area and a frozen embryo transferring area, wherein two sides of the embryo loading area are respectively communicated with the frozen dripping area and the frozen embryo transferring area, and the embryo loading area is at the lowest position. By designing three accommodating areas, wherein the embryo loading area is designed at the lowest position and is used for placing and positioning embryos, the liquid is prevented from flowing in the freezing process, and the embryos are driven to move; the frozen drip zone is used for the drip and removal of balancing liquid and cryoprotectant while ensuring that less liquid remains in the embryo loading zone when removed; the thawing embryo transfer zone is used to allow the embryos to fall off more easily during thawing, while ensuring that less liquid remains in the embryo loading zone during the freezing process.
In the above-described aspect, the embryo loading area is a concave downward structure or an inverted cone structure.
In the above-mentioned scheme, the side of embryo loading district with frozen drip district links to each other is the vertical face. By designing one side of the embryo loading area connected with the frozen dripping area as a vertical surface, on one hand, the embryos can be effectively prevented from sliding to the frozen dripping area in the operation process, so that the safety of the embryos is ensured; on the other hand, the volume of the embryo loading area can be greatly reduced, so that the residual liquid during liquid removal and before embryo freezing is ensured to be as little as possible.
In the above scheme, the side of the embryo loading area connected with the thawing embryo transferring area is an inclined plane or an arc surface. By designing the side of the embryo loading area connected with the thawing embryo transferring area as an inclined plane or an arc surface, on one hand, embryos can fall off more easily during thawing, and on the other hand, liquid can flow more easily, so that less liquid remains in the embryo loading area.
In the above embodiment, the embryo loading area has a volume of 0.1 to 0.9ul.
In the above scheme, the side of the frozen drip area, which is communicated with the embryo loading area, is of a planar structure. When in use, the plane structure of the frozen liquid dropping area is horizontally arranged. By designing the side of the frozen drip area which is in communication with the embryo loading area as a planar structure, drip and removal of liquid is facilitated.
In the above scheme, one side of the frozen drip area far away from the embryo loading area is of an arc surface structure. Through the design of one side of freezing dropping liquid district far away from embryo loading district as the cambered surface structure, can increase the operating space of dropping liquid buret greatly like this to the operation has been made things convenient for.
In the above scheme, the junction of the planar structure and the cambered surface structure of the frozen liquid dropping area is arc-shaped. Because the head of the drip-dispensing tube is of a circular plane structure, the joint of the plane structure of the frozen drip-dispensing region and the cambered surface structure is designed to be circular arc, so that the head of the drip-dispensing tube can be better contacted with the head of the drip-dispensing tube, meanwhile, the liquid is convenient to remove because the corners of the circular arc are not easy to remain, and the circular arc is also favorable for production and processing.
In the above scheme, the cambered surface structure of the frozen drip zone is inclined outwards away from one side of the embryo loading zone. Through the cambered surface structure that keeps away from embryo loading district with freezing drip district one side design outside slope, can further increase the operating space of drip buret like this to convenient operation, liquid overflow when also can prevent the operation simultaneously.
In the above scheme, two sides of the embryo loading area are respectively connected with the frozen dripping area and the thawing embryo transferring area in a smooth transition manner. The connection part of the embryo loading area and the frozen dripping area and the unfreezing embryo transferring area is subjected to smooth treatment, so that on one hand, the flow of liquid is convenient, and less liquid remains in the embryo loading area; on the other hand, when thawing, the embryo can be more easily dropped and no scratch is generated on the embryo.
In the above scheme, the thawing embryo transfer region has an arc surface structure. Through designing the thawing embryo transferring area as an arc surface structure, the embryo can be further easily dropped off during thawing, and the embryo can be quickly separated from the freezing carrying rod.
In the above-mentioned scheme, the lowest height of the thawing embryo transferring region is greater than or equal to the height of the planar structure of the frozen drip region, and the height of the planar structure of the frozen drip region is greater than or equal to the highest height of the embryo loading region. The minimum height of the thawing embryo transferring area is designed to be larger than or equal to the height of the plane structure of the frozen dripping area, and the height of the plane structure of the frozen dripping area is designed to be larger than or equal to the maximum height of the embryo loading area, so that liquid is convenient to remove, and residual liquid before embryo freezing is ensured to be as small as possible.
In the above aspect, the lateral dimension D1 of the embryo loading area is smaller than the lateral dimension D2 of the frozen drip area, and the lateral dimension D1 of the embryo loading area is smaller than the lateral dimension D3 of the thawed embryo transfer area. By designing the lateral dimension D1 of the embryo loading area to be minimal, the volume of the embryo loading area can be further reduced, thereby further ensuring that as little liquid as possible remains during liquid removal and before embryo freezing.
In the above-described aspect, the lateral dimension D3 of the thawed embryo transfer area is smaller than the lateral dimension D2 of the frozen drip area. By designing the lateral dimension D3 of the thawing embryo transfer region to be smaller than the lateral dimension D2 of the frozen drip region, this facilitates rapid immersion of the frozen carrier bar into liquid nitrogen.
In the above scheme, an operating handle is arranged on the outer wall of the frozen drip area.
In the above scheme, the volume of the frozen dripping zone is 8-14ul, and the volume of the thawing embryo transferring zone is 8-14ul.
A method for operating a frozen carrier bar comprises the following steps,
s1, transferring an embryo into an embryo loading area;
s2, removing redundant culture solution;
s3, adding balance liquid into the frozen liquid dropping area to submerge the embryo with the balance liquid;
s4, allowing the embryo to stand in the balance liquid for T1 min;
s5, removing the balance liquid;
s6, dripping the cryoprotectant into the freezing drip area to submerge the embryo with the cryoprotectant;
s7, allowing the embryo to stand in a cryoprotectant for T2 min;
s8, removing the cryoprotectant;
s9 submerging the frozen carrier bar in liquid nitrogen.
In the process of using the freezing carrier rod to carry out freezing operation, only one transfer operation is needed to be carried out on the embryo, compared with the freezing carrier rod used in the existing freezing operation, the embryo transfer times are reduced, and the damage to the embryo possibly caused by transfer is reduced. Moreover, since only one transfer operation is required for the embryo, the automated design of the operation is facilitated.
In the above protocol, the embryo is allowed to stand in the equilibration solution for 5-10 minutes and the embryo is allowed to stand in the cryoprotectant for 0.5-1.5 minutes.
The invention provides a freezing carrying rod and an operation method thereof. The beneficial effects are as follows:
1. by designing three accommodating areas, wherein the embryo loading area is designed at the lowest position and is used for placing and positioning embryos, the liquid is prevented from flowing in the freezing process, and the embryos are driven to move; the frozen drip zone is used for the drip and removal of balancing liquid and cryoprotectant while ensuring that less liquid remains in the embryo loading zone when removed; the embryo transferring area is used for enabling the embryos to fall off more easily during thawing, and ensuring that less liquid remains in the embryo loading area during freezing;
2. by designing one side of the embryo loading area connected with the frozen dripping area as a vertical surface, on one hand, the embryos can be effectively prevented from sliding to the frozen dripping area in the operation process, so that the safety of the embryos is ensured; on the other hand, the volume of the embryo loading area can be greatly reduced, so that the residual liquid during liquid removal and before embryo freezing is ensured to be as little as possible;
3. by designing one side of the embryo loading area connected with the thawing embryo transferring area as an inclined plane or an arc surface, on one hand, embryos can fall off more easily during thawing, and on the other hand, liquid can flow more easily, so that less liquid remains in the embryo loading area;
4. when in use, the plane structure of the frozen liquid dropping area is horizontally arranged. By designing the side, communicated with the embryo loading area, of the frozen dripping area as a planar structure, the dripping and removing of liquid are facilitated;
5. by designing one side of the frozen drip area far away from the embryo loading area as an arc surface structure, the operation space of the drip burette can be greatly increased, thereby facilitating the operation;
6. the head of the drip-dispensing tube is of a circular structure, so that the joint of the planar structure of the frozen drip area and the cambered surface structure is designed to be arc-shaped, the head of the drip-dispensing tube can be better matched with the head of the drip-dispensing tube, meanwhile, the liquid is convenient to remove because the corners of the arc-shaped are not easy to remain, and the arc-shaped structure is also beneficial to production and processing;
7. the cambered surface structure of the frozen drip area is designed to be inclined outwards at one side far away from the embryo loading area, so that the operation space of the drip burette can be further increased, the operation is convenient, and meanwhile, liquid overflow during the operation can be prevented;
8. the connection part of the embryo loading area and the frozen dripping area and the unfreezing embryo transferring area is subjected to smooth treatment, so that on one hand, the flow of liquid is convenient, and less liquid remains in the embryo loading area; on the other hand, when thawing, the embryo can be more easily dropped off and no scratch can be generated on the embryo;
9. the thawing embryo transferring area is designed to be of an arc surface structure, so that the embryo can be further easily separated during thawing, and the embryo can be quickly separated from the freezing carrying rod;
10. the minimum height of the thawing embryo transferring area is designed to be greater than or equal to the height of the plane structure of the frozen dripping area, and the height of the plane structure of the frozen dripping area is designed to be greater than or equal to the maximum height of the embryo loading area, so that liquid is convenient to remove, and residual liquid before embryo freezing is ensured to be as little as possible;
11. by designing the lateral dimension D1 of the embryo loading area to be minimal, the volume of the embryo loading area can be further reduced, thereby further ensuring that as little liquid as possible remains during liquid removal and before embryo freezing;
12. the transverse dimension D3 of the thawing embryo transferring region is designed to be smaller than the transverse dimension D2 of the frozen dripping region, so that the frozen carrying rod is conveniently and rapidly immersed into liquid nitrogen;
13. in the process of using the freezing carrying rod to carry out freezing operation, only one transfer operation is needed to be carried out on the embryo, compared with the freezing carrying rod used in the existing freezing operation, the embryo transfer times are reduced, the damage to the embryo possibly caused by transfer is reduced, and in addition, the automatic design of the operation is facilitated because the embryo is only needed to be transferred once.
Drawings
FIG. 1 is a schematic view of the structure of the present frozen carrier bar;
FIG. 2 is a schematic cross-sectional view of FIG. 1;
FIG. 3 is a schematic view of a partial enlarged structure of FIG. 2;
fig. 4 is a partially enlarged schematic structural view of fig. 1.
In the figure: embryo loading area 1, frozen drip area 2, thawing embryo transfer area 3, and operating handle 4.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
As shown in fig. 1-4, the present invention provides a technical solution: the frozen carrying rod comprises an embryo loading area 1, a frozen dripping area 2 and a frozen embryo transferring area 3, wherein two sides of the embryo loading area 1 are respectively communicated with the frozen dripping area 2 and the frozen embryo transferring area 3, and the embryo loading area 1 is at the lowest position. By designing three accommodating areas, wherein the embryo loading area 1 is designed at the lowest position and is used for placing and positioning embryos, the flow of liquid in the freezing process is prevented, and the embryos are driven to move; the frozen drip zone 2 is used for the drip and removal of balancing liquid and cryoprotectant, while ensuring that less liquid remains in the embryo loading zone 1 when removed; the thawing embryo transfer zone 3 is used to allow the embryos to fall off more easily during thawing, while ensuring that less liquid remains in the embryo loading zone 1 during the freezing process.
The embryo loading area 1 is of a concave or inverted cone-shaped configuration, downwards. The side of the embryo loading area 1 connected with the frozen dripping area 2 is a vertical surface. By designing the side, connected with the embryo loading area 1 and the frozen dripping area 2, as a vertical surface, on one hand, embryos can be effectively prevented from sliding to the frozen dripping area 2 in the operation process, so that the safety of the embryos is ensured; on the other hand, the volume of the embryo loading area 1 can be greatly reduced, thereby ensuring that as little liquid as possible remains during liquid removal and before embryo freezing. One side of the embryo loading area 1 connected with the thawing embryo transferring area 3 is an inclined plane or an arc surface. By designing the side of the embryo loading area 1 connected with the thawing embryo transferring area 3 as an inclined plane or an arc surface, on the one hand, embryos can be more easily separated during thawing, and on the other hand, liquid can be more easily flowed, so that less liquid remains in the embryo loading area 1. The embryo loading zone 1 has a volume of 0.1-0.9ul.
The side of the frozen drip area 2 communicated with the embryo loading area 1 is a plane structure. In use, the planar structure of the refrigerated drip zone 2 is arranged horizontally. By designing the side of the frozen drip area 2 that communicates with the embryo loading area 1 as a planar structure, drip and removal of liquid is facilitated. The side of the frozen drip area 2 far away from the embryo loading area 1 is of an arc surface structure. By designing the side of the frozen drip area 2, which is far away from the embryo loading area 1, as an arc surface structure, the operating space of the drip burette can be greatly increased, thereby facilitating the operation. The junction of the planar structure and the cambered surface structure of the frozen liquid dropping area 2 is arc-shaped. Because the head of the drip-dispensing tube is of a circular structure, the joint of the planar structure of the frozen drip region 2 and the cambered surface structure is designed to be circular arc, so that the head of the drip-dispensing tube can be better matched with the head of the drip-dispensing tube, meanwhile, the liquid is convenient to remove because the corners of the circular arc are not easy to remain, and the circular arc is also favorable for production and processing. The cambered surface structure of the frozen drip zone 2 is inclined outwards away from one side of the embryo loading zone 1. The cambered surface structure of the frozen drip area 2 is designed to be inclined outwards away from one side of the embryo loading area 1, so that the operation space of the drip burette can be further increased, the operation is convenient, and meanwhile, liquid overflow during the operation can be prevented.
The two sides of the embryo loading area 1 are respectively connected with the frozen dripping area 2 and the unfreezing embryo transferring area 3 in a smooth transition way. The connection part between the embryo loading area 1 and the frozen dripping area 2 and the unfreezing embryo transferring area 3 is subjected to smooth treatment, so that on one hand, the flow of liquid is convenient, and less liquid remains in the embryo loading area 1; on the other hand, when thawing, the embryo can be more easily dropped and no scratch is generated on the embryo. The thawing embryo transferring area 3 is in a cambered surface structure. Through designing the thawing embryo transferring area 3 as an arc surface structure, the embryo can be further easily dropped off during thawing, and the embryo can be quickly separated from the frozen carrying rod.
The lowest height of the thawing embryo transferring zone 3 is greater than or equal to the height of the planar structure of the frozen dripping zone 2, and the height of the planar structure of the frozen dripping zone 2 is greater than or equal to the highest height of the embryo loading zone 1. The minimum height of the thawing embryo transferring area 3 is designed to be larger than or equal to the height of the plane structure of the freezing drip area 2, and the height of the plane structure of the freezing drip area 2 is designed to be larger than or equal to the highest height of the embryo loading area 1, so that liquid is convenient to remove, and residual liquid before embryo freezing is ensured to be as small as possible.
The lateral dimension D1 of the embryo loading area 1 is smaller than the lateral dimension D2 of the frozen drip area 2, and the lateral dimension D1 of the embryo loading area 1 is smaller than the lateral dimension D3 of the thawing embryo transfer area 3. By designing the lateral dimension D1 of the embryo loading area 1 to be minimal, the volume of the embryo loading area 1 can be further reduced, thereby further ensuring that as little liquid as possible remains during liquid removal and before embryo freezing. The transverse dimension D3 of the thawing embryo transfer region 3 is smaller than the transverse dimension D2 of the frozen drip region 2. By designing the lateral dimension D3 of the thawing embryo transfer region 3 to be smaller than the lateral dimension D2 of the frozen drip region 2, this facilitates rapid immersion of the frozen carrier bar into liquid nitrogen.
An operating handle 4 is arranged on the outer wall of the frozen drip area 2. The volume of the frozen dripping zone 2 is 8-14ul, and the volume of the thawing embryo transferring zone 3 is 8-14ul.
In the embodiment, three accommodating areas are designed, wherein the embryo loading area 1 is designed at the lowest position and is used for placing and positioning embryos, so that liquid is prevented from flowing in the freezing process, and the embryos are driven to move; the frozen drip zone 2 is used for the drip and removal of balancing liquid and cryoprotectant, while ensuring that less liquid remains in the embryo loading zone 1 when removed; the thawing embryo transfer zone 3 is used to allow the embryos to fall off more easily during thawing, while ensuring that less liquid remains in the embryo loading zone 1 during the freezing process.
Meanwhile, in the embodiment, one side, connected with the embryo loading area 1 and the frozen dripping area 2, is designed to be a vertical surface, so that on one hand, embryos can be effectively prevented from sliding to the frozen dripping area 2 in the operation process, and the safety of the embryos is ensured; on the other hand, the volume of the embryo loading area 1 can be greatly reduced, so that the residual liquid during liquid removal and before embryo freezing is ensured to be as little as possible; by designing the side, connected with the embryo loading area 1 and the thawing embryo transferring area 3, as an inclined plane or an arc surface, on one hand, embryos can fall off more easily during thawing, and on the other hand, liquid can flow more easily, so that less liquid remains in the embryo loading area 1;
moreover, in the embodiment, the side, which is communicated with the embryo loading area 1, of the frozen dripping area 2 is designed to be of a plane structure, so that the dripping and removing of liquid are convenient; by designing the side of the frozen drip area 2 far away from the embryo loading area 1 as an arc surface structure, the operation space of the drip burette can be greatly increased, thereby facilitating the operation; the head of the drip tube is of a circular structure, so that the joint of the planar structure of the frozen drip region 2 and the cambered surface structure is designed to be arc-shaped, the head of the drip tube can be better matched with the head of the drip tube, meanwhile, the liquid is not easy to be removed due to the fact that the corners of the arc-shaped are not easy to remain, and the arc-shaped structure is also favorable for production and processing; the cambered surface structure of the frozen drip area 2 is designed to be inclined outwards away from one side of the embryo loading area 1, so that the operation space of the drip burette can be further increased, the operation is convenient, and meanwhile, liquid overflow during the operation can be prevented.
In addition, in the embodiment, the connection part between the embryo loading area 1 and the frozen dripping area 2 and the unfreezing embryo transferring area 3 is subjected to smoothing treatment, so that on one hand, the flow of liquid is convenient, and less liquid remains in the embryo loading area 1; on the other hand, when thawing, the embryo can be more easily dropped off and no scratch can be generated on the embryo; through designing the thawing embryo transferring area 3 as an arc surface structure, the embryo can be further easily dropped off during thawing, and the embryo can be quickly separated from the frozen carrying rod.
Finally, in this embodiment, the minimum height of the thawing embryo transferring region 3 is designed to be greater than or equal to the height of the planar structure of the frozen dripping region 2, and the height of the planar structure of the frozen dripping region 2 is designed to be greater than or equal to the maximum height of the embryo loading region 1, so that the liquid is convenient to remove, and the residual liquid before freezing of the embryo is ensured to be as small as possible; by designing the lateral dimension D1 of the embryo loading area 1 to be minimal, the volume of the embryo loading area 1 can be further reduced, thereby further ensuring that as little liquid as possible remains during liquid removal and before embryo freezing; by designing the lateral dimension D3 of the thawing embryo transfer region 3 to be smaller than the lateral dimension D2 of the frozen drip region 2, this facilitates rapid immersion of the frozen carrier bar into liquid nitrogen.
Example 2
As shown in fig. 1-4, the present invention provides a technical solution: a method for operating a frozen carrier bar comprises the following steps,
s1, transferring embryos together with culture solution into a frozen drip area 2 of the frozen carrier rod;
s2, removing redundant culture solution by using a Pasteur tube;
s3, sucking 20ul of balance liquid by using a Pasteur pipe, and dripping the balance liquid into the frozen drip area 2 so that the balance liquid can completely submerge the embryo;
s4, allowing the embryo to stand in the balance liquid for 5 minutes;
s5, removing the balance liquid by using a Pasteur pipe;
s6, repeating the steps S3-S5 for a plurality of times if needed;
s7, sucking 20ul of cryoprotectant by using a Pasteur pipe, and dripping the cryoprotectant into the freezing drip area 2 so that the cryoprotectant can completely submerge the embryo;
s8, allowing the embryo to stand in a cryoprotectant for 1 minute;
s9, removing the cryoprotectant by using a Pasteur pipe;
s10, repeating S6-S8 for a plurality of times if needed;
s11, quickly immersing the freezing carrier rod into liquid nitrogen.
As can be seen from the above operation steps, in the process of freezing operation by using the freezing carrier rod, only one transfer operation is needed to be carried out on the embryo, compared with the freezing carrier rod used in the existing freezing operation, the embryo transfer times are reduced, and the damage and embryo loss possibly caused by transfer are reduced. Moreover, since only one transfer operation is required for the embryo, the automated design of the operation is facilitated.
Claims (13)
1. A frozen carrier bar, characterized in that: the embryo transfer device comprises an embryo loading area (1), a frozen dripping area (2) and a thawing embryo transfer area (3), wherein two sides of the embryo loading area (1) are respectively communicated with the frozen dripping area (2) and the thawing embryo transfer area (3), and the embryo loading area (1) is at the lowest position;
the embryo loading area (1) is of a downward concave structure or an inverted cone structure; one side of the embryo loading area (1) connected with the frozen dripping area (2) is a vertical surface; one side of the embryo loading area (1) connected with the thawing embryo transferring area (3) is an inclined plane or an arc surface; two sides of the embryo loading area (1) are respectively connected with the frozen dripping area (2) and the unfreezing embryo transferring area (3) in a smooth transition manner; the lowest height of the thawing embryo transferring area (3) is larger than or equal to the height of the plane structure of the frozen dripping area (2), and the height of the plane structure of the frozen dripping area (2) is larger than or equal to the highest height of the embryo loading area (1).
2. A frozen carrier bar as defined in claim 1, wherein: the embryo loading zone (1) has a volume of 0.1-0.9ul.
3. A frozen carrier bar as defined in claim 2, wherein: the side of the frozen liquid dropping area (2) communicated with the embryo loading area (1) is of a plane structure.
4. A frozen carrier bar as defined in claim 3, wherein: one side of the frozen liquid dropping area (2) far away from the embryo loading area (1) is of an arc surface structure.
5. A frozen carrier bar as defined in claim 4, wherein: the junction of the planar structure and the cambered surface structure of the frozen liquid dropping area (2) is arc-shaped.
6. A frozen carrier bar as recited in claim 5, wherein: the cambered surface structure of the frozen drip zone (2) is inclined outwards away from one side of the embryo loading zone (1).
7. A frozen carrier bar as defined in claim 6, wherein: the thawing embryo transferring area (3) is of an arc surface structure.
8. A frozen carrier bar as defined in claim 7, wherein: the lateral dimension D1 of the embryo loading area (1) is smaller than the lateral dimension D2 of the frozen dripping area (2), and the lateral dimension D1 of the embryo loading area (1) is smaller than the lateral dimension D3 of the thawing embryo transferring area (3).
9. A frozen carrier bar as defined in claim 8, wherein: the transverse dimension D3 of the thawing embryo transfer region (3) is smaller than the transverse dimension D2 of the frozen dripping region (2).
10. A frozen carrier bar as defined in claim 1, wherein: an operating handle (4) is arranged on the outer wall of the frozen liquid dropping area (2).
11. A frozen carrier bar as defined in claim 1, wherein: the volume of the frozen dripping zone (2) is 8-14ul, and the volume of the thawing embryo transferring zone (3) is 8-14ul.
12. A method of operating a frozen carrier bar according to claim 9, characterized in that: comprises the steps of,
s1 transferring an embryo into an embryo loading area (1);
s2, removing redundant culture solution;
s3, adding balance liquid into the frozen liquid dropping area (2) to submerge the embryo with the balance liquid;
s4, allowing the embryo to stand in the balance liquid for T1 min;
s5, removing the balance liquid;
s6, dripping the cryoprotectant into the freezing drip area (2) to submerge the embryo with the cryoprotectant;
s7, allowing the embryo to stand in a cryoprotectant for T2 min;
s8, removing the cryoprotectant;
s9 submerging the frozen carrier bar in liquid nitrogen.
13. A method of operating a frozen carrier bar as recited in claim 12 wherein: the embryo is kept stand in the balancing solution for 5-10 min, and the embryo is kept stand in the cryoprotectant for 0.5-1.5 min.
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| CN202110943874.0A CN113632785B (en) | 2021-08-17 | 2021-08-17 | Freezing carrying rod and operation method thereof |
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| CN208291916U (en) * | 2018-06-01 | 2018-12-28 | 成都艾伟孚生物科技有限公司 | A kind of mouse embryo means of transport |
| CN216135089U (en) * | 2021-08-17 | 2022-03-29 | 武汉互创联合科技有限公司 | Freezing pole that carries |
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2021
- 2021-08-17 CN CN202110943874.0A patent/CN113632785B/en active Active
Patent Citations (6)
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| WO2013098825A1 (en) * | 2012-01-01 | 2013-07-04 | A.A. Cash Technology Ltd | Cryopreservation device for vitrification and method for using same |
| CN103749436A (en) * | 2014-01-23 | 2014-04-30 | 江苏苏云医疗器材有限公司 | Enclosed type embryo freezing carrying rod as well as preparation method and use method thereof |
| CN105707057A (en) * | 2016-04-19 | 2016-06-29 | 戴志俊 | Method for unfreezing gamete or embryo on frozen carrier and vessel special for vitrification unfreezing |
| CN207519501U (en) * | 2017-10-16 | 2018-06-22 | 山东威高新生医疗器械有限公司 | Freeze straw |
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| CN113632785A (en) | 2021-11-12 |
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