CN107022484A - A kind of Embryo Culture unit and preparation method thereof - Google Patents
A kind of Embryo Culture unit and preparation method thereof Download PDFInfo
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- CN107022484A CN107022484A CN201710390586.0A CN201710390586A CN107022484A CN 107022484 A CN107022484 A CN 107022484A CN 201710390586 A CN201710390586 A CN 201710390586A CN 107022484 A CN107022484 A CN 107022484A
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- 210000001161 mammalian embryo Anatomy 0.000 title claims abstract description 168
- 238000002360 preparation method Methods 0.000 title claims description 6
- 238000005070 sampling Methods 0.000 claims abstract description 28
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 26
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 13
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 239000012780 transparent material Substances 0.000 claims abstract description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 23
- 239000000741 silica gel Substances 0.000 claims description 23
- 229910002027 silica gel Inorganic materials 0.000 claims description 23
- 238000000465 moulding Methods 0.000 claims description 14
- 230000007704 transition Effects 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 10
- 231100000252 nontoxic Toxicity 0.000 claims description 7
- 230000003000 nontoxic effect Effects 0.000 claims description 7
- 238000005266 casting Methods 0.000 claims description 3
- 230000008602 contraction Effects 0.000 claims description 3
- 230000003628 erosive effect Effects 0.000 claims description 3
- 238000009738 saturating Methods 0.000 claims description 2
- 238000011534 incubation Methods 0.000 abstract description 7
- 238000000338 in vitro Methods 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 210000002459 blastocyst Anatomy 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 81
- 238000007789 sealing Methods 0.000 description 8
- 230000008859 change Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 6
- 238000005452 bending Methods 0.000 description 5
- 239000002253 acid Substances 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 210000004681 ovum Anatomy 0.000 description 3
- 238000010146 3D printing Methods 0.000 description 2
- 206010038743 Restlessness Diseases 0.000 description 2
- 238000004891 communication Methods 0.000 description 2
- 230000007797 corrosion Effects 0.000 description 2
- 238000005260 corrosion Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000013020 embryo development Effects 0.000 description 2
- 210000002308 embryonic cell Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000007493 shaping process Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000009191 jumping Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
Classifications
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- 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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
-
- 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
- C12M23/12—Well or multiwell plates
-
- 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
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/04—Seals
Abstract
The invention belongs to Embryo Culture technical field, it is related to a kind of Embryo Culture unit, culture unit main body including transparent material, the upper surface of culture unit main body opens up a sample introduction end plug hole and a sampling end consent, plug seal is filled in sample introduction end plug hole and sampling end consent, the bottom in sample introduction end plug hole sets sample holes, the bottom of sampling end consent sets thief hole, the underface of thief hole sets embryo's accommodating chamber, cultivate and Embryo Culture chamber is set in unit main body, Embryo Culture chamber connects sample holes by sample intake passage, Embryo Culture chamber connects embryo's accommodating chamber by sample output passage.This Embryo Culture unit loads with after the good formula culture medium of carbon dioxide pre-balance, constitute the In vitro culture system of an absolute tightness, embryonated egg ectogenesis can be supported to blastocyst stage directly in 37 DEG C of common insulating boxs, without CO2gas incubator, it is easy to observe the development of embryo, incubation step is significantly simplified, operating efficiency and quality is improved.
Description
Technical field
The invention belongs to Embryo Culture fixture technology field, it is related to a kind of Embryo Culture unit and preparation method thereof.
Background technology
Current inseminatio externalis and Embryo Culture are completed in CO2gas incubator, rely on temperature constant in incubator
The carbon dioxide of degree and constant density interacts and exchanged with the chemical composition in culture medium, to maintain smart ovum to inseminate and embryo
Physiological environment needed for growth.But either on hardware device (culture apparatus or incubator) or exist in existing culture systems
All there is some defects or deficiency in terms of culture medium.For hardware, existing culture apparatus (incubator) is one open
System, the gas in CO2gas incubator needed constantly with being that the air in embryo experiments room exchange and could maintain to train outside case
Support in case or in device gas balance and stability, therefore there is following clearly disadvantageous or defect in such open system：
One is that gas concentration and temperature in device are actually to be in a kind of dynamic poised state, and gas concentration and temperature are certain
In the range of fluctuate, the size of fluctuation then depends on the enabling frequency of model, quality and the incubator of device or incubator；Two are
Volatile organic matter in laboratory, harmful physical and chemical factor such as ozone etc. can pass through this open gas communication system
Enter in CO2gas incubator, cause the damage of embryo, in addition it is dead；The third is when CO2gas incubator opens the door,
Gas in case largely overflows and fluctuated with inevitably resulting in the gas concentration in incubator and high temperature, with enabling
The increase of number of times, the degree that this big ups and downs cause embryo to injure also increases therewith.
The content of the invention
It is an object of the invention to provide a kind of Embryo Culture unit, it is previously charged into when Embryo Culture unit is made
With the good culture medium of carbon dioxide pre-equilibration, adding embryonated egg or embryo can cultivate to blastaea or required embryonic development rank
Section, when using only need in 37 DEG C of insulating boxs rewarming, without in CO2gas incubator balance, observation embryo when also without
Embryo need to be taken out, whole device need to only be taken out, be placed in the micro- Microscopic observation with temperature-constant plate or take pictures.During observation not
The microenvironment of Embryo Culture can be caused to produce any change or fluctuation, the microenvironment of embryo growth is protected in whole incubation
Hold constant.
To achieve these goals, present invention employs following technical proposals.A kind of Embryo Culture unit, including transparent material
The culture unit main body of matter, the upper surface of culture unit main body opens up a sample introduction end plug hole and a sampling end consent, sample introduction
Plug seal is filled in end plug hole and sampling end consent, the bottom in sample introduction end plug hole sets sample holes, the bottom of sampling end consent
Thief hole is set, and the underface of thief hole, which is set in embryo's accommodating chamber, culture unit main body, sets Embryo Culture chamber, Embryo Culture
Chamber connects sample holes by sample intake passage, and Embryo Culture chamber connects embryo's accommodating chamber by sample output passage.
A kind of preferably technical scheme, the most narrow place of sample intake passage is located at upper end, and sample holes are gradually tapered up, seamlessly transit to
The most narrow place of sample intake passage, then sample intake passage gradually amplify, sample intake passage lower end is connected with Embryo Culture chamber；Sample output passage
Lower end connects Embryo Culture chamber, and sample output passage is gradually tapered up, and gradually amplifies again after the most narrow place of sample output passage, using smooth circle
Arc transition is connected with embryo's accommodating chamber.It is designed to significantly reduce at both ends open by slot indirect in short-term during plug due to opening
Tactile air may cause the minor fluctuations of Embryo Culture intracavitary nutrient solution chemical factors, so as to farthest ensure culture ring
The constancy in border.
The junction of a kind of preferably technical scheme, Embryo Culture chamber and sample intake passage is by rapid desufflation section transition, soon
Fast contraction section, which is played, prevents embryonated egg or embryo to be back to sample intake passage.
A kind of preferably technical scheme, sample intake passage is connected with the junction of Embryo Culture chamber by one section of transition arc,
The bending direction and Embryo Culture cavity wall bending direction of the bending direction of transition arc and the sample intake passage wall of the junction on the contrary,
Certain transition arc should be tangent with sample intake passage and Embryo Culture chamber, to ensure that embryonated egg smoothly flows into Embryo Culture chamber.
A kind of preferably technical scheme, the narrowest diameter of sample intake passage is not more than 1.5mm, and the most narrow place of sample output passage leads to
Road diameter is 1.5-2.0mm.
A kind of preferably technical scheme, from sample holes to sample intake passage, then to Embryo Culture chamber, then to sample output passage, then
Whole vias inner walls to embryo's accommodating chamber are seamlessly transitted and smooth, and without turning and burr, corner is used
Tangent arc transition so that embryonated egg or embryonic cell slidably, are not tangled.
A kind of preferably technical scheme, culture unit main body uses glass material, it would however also be possible to employ other are arbitrarily to embryo
Nontoxic transparent material, plug uses silica gel plug, and silica gel plug elasticity is good, is easy to sealing, and nontoxic to embryo.
A kind of preferably technical scheme, the bottom of Embryo Culture chamber is curved, it is ensured that in incubation, embryonated egg or embryo
Pool together, and rest on the lowest part of Embryo Culture chamber.
A kind of preferably technical scheme, the bottom of embryo's accommodating chamber is curved, and the minimum point of embryo's accommodating chamber is located at
The underface of thief hole, conveniently takes embryo.
The horizontal fin of multiple tracks, can be adopted on the side wall of a kind of preferably technical scheme, sample introduction end plug hole and sampling end consent
With shape of threads, or thread like fin, horizontal fin can extrude silica gel plug, play sealing effectiveness, it is ensured that air tight.
A kind of preferably technical scheme, the aperture outer of sample holes is higher than the base plane in sample introduction end plug hole so that aperture
Outer can squeeze into silica gel plug bottom surface, it is ensured that sealing effectiveness.Similarly, the aperture outer of thief hole is higher than the bottom of sampling end consent
Plane.
A kind of a diameter of 10mm of preferably technical scheme, sample introduction end plug hole and sampling end consent.Sample holes are diameter 4mm
It is round, thief hole is ellipse, and optimal thief hole is major axis 6mm, short axle 3mm ellipse.
A kind of preferably technical scheme, the surface of culture unit main body sets breach, and breach is used to fix elastic band, elastic
The fixed silica gel plug of band, prevents silica gel plug from loosening, or have other structures for preventing plug from surprisingly loosening.
A kind of preferably technical scheme, culture unit main body upper surface sets the neck for placing silica gel plug, in order to extract
Fixed placement after silica gel plug, unrest is not mixed up or is dropped.
A kind of foregoing Embryo Culture unit manufacturing process is：Make first and the sample introduction end plug inside culture unit main body
The consistent integral inside molding of hole, sample holes, sample intake passage, Embryo Culture chamber, embryo's accommodating chamber, thief hole, sampling end consent shape,
Fixed integral inside molding, moulding by casting culture unit main body, then erosion removal integral inside molding again, so as to obtain Embryo Culture list
Member.When 3D printing technique is improved enough, this Embryo Culture unit directly can also be made using 3D printing technique.
Further, to the injection of Embryo Culture intracavitary, using the good embryo culture medium of carbon dioxide balance, (recommendation makes in advance
With a formula culture medium), then seal sample introduction end plug hole and sampling end consent with silica gel plug.
Further, the material selection metal of the integral inside molding is preferred, and is corroded using chemical substance or acid.Culture
The material of unit main body is using glass or other arbitrarily easily shapings, and nontoxic to embryo, and not by the material of acid corrosion.
Advantages of the present invention：1. it is previously implanted in advance using the good embryo culture medium of carbon dioxide balance, after sealing of jumping a queue
Embryo as instant independently cultivates unit, user in use, culture unit is directly added after 37 DEG C of rewarmings from sample holes
The embryo of fertilised egg or different developmental phases, and be positioned in common insulating box and cultivate to the required stage of development.
Operate extremely convenient, it is simple, not only significantly reduce working strength, improve operating efficiency, and considerably reduce operation
The risk of error；2. user, without using carbon dioxide balance culture medium again, significantly reduces Embryo Culture room pair using preceding
The usage amount of CO2gas incubator and the usage amount of carbon dioxide, save expense and improve culturing room's security again；
3. due to Embryo Culture unit volume very little, preferably volume is 55mm*25mm*35mm (the long wide * of * are high), a normal size
Insulating box can at least place 200 Embryo Culture units, by every CO2gas incubator simultaneously cultivate be no more than two
If the recommendation usage of people's embryo, if using this Embryo Culture unit, the culture capacity phase of a standard size insulating box
When in 100 use capacity with the CO2gas incubator under conventional culture conditions, therefore substantial amounts of experiment is not only saved
Room space, carbon dioxide, also improve the security of embryo experiments room；4. Embryo Culture unit is one independent closed
Temperature and pH value in culture systems, culture chamber can keep the constancy of height, will not become as constant incubator opens the door
Change, will not be interfered with each other between different cultures, any variation of the external environment in addition to culturing room powers off for a long time or thing
Therefore (such as ultraviolet light of laboratory sterilization) does not interfere with cultivation result, Embryo Culture quality is not interfered with, so that significantly
Degree improves the training quality of embryo, and in vitro culture safety and stability.Even if powering off in addition, it will can also cultivate
Unit is taken out, and culture is continued to by body temperature, and this characteristic is also convenient for the emergent transfer or transport of embryo；5. add fertilization
After ovum, incubation can be intuitively observed, camera is installed, incubation can be dynamically observed；When taking out observation in vitro, also will not
Change the chemical factors in culture chamber, this characteristic is for extremely crucial for the embryo sensitive to environmental change；6. from embryo
When accommodating chamber takes embryo, as found to still need to culture when taking embryo, embryo can be made to be back to Embryo Culture chamber and continue to cultivate, and returned
Embryo will not flow backward to sample intake passage during stream；7. whole Embryo Culture unit good airproof performance, sample intake passage and sample output passage are used
Contraction type structure, and seamlessly transit, air exclusion is good；8. the preparation method of Embryo Culture unit solves processing inner chamber
The problem of body passage, it is ensured that interior smooth impulse- free robustness, and transitional region circular arc seamlessly transits.
Brief description of the drawings
Fig. 1 is the Embryo Culture cell schematics of the present invention.
Embodiment
The present invention is further clarified below in conjunction with the accompanying drawings.
Embodiment 1
A kind of reference picture 1, Embryo Culture unit includes the culture unit main body 1 of transparent material, culture unit main body 1
Upper surface opens up in a sample introduction end plug hole 3 and a sampling end consent 4, sample introduction end plug hole 3 and sampling end consent 4 and fills in plug
Sealing, the bottom in sample introduction end plug hole 3 sets sample holes 5, and the bottom of sampling end consent 4 sets thief hole 6, thief hole 6 just under
Side is set in embryo's accommodating chamber 7, culture unit main body 1 and sets Embryo Culture chamber 2, and Embryo Culture chamber 2 is connected by sample intake passage 8
Sample holes 5, Embryo Culture chamber 2 connects embryo's accommodating chamber 7 by sample output passage 9.
In the present embodiment, culture unit main body 1 uses glass material, it would however also be possible to employ other are arbitrarily nontoxic to embryo saturating
Bright material, plug uses silica gel plug, and silica gel plug elasticity is good, is easy to sealing, and nontoxic to embryo.
In the present embodiment, the narrowest diameter of sample intake passage 8 is not more than in 1.5mm, actual processing, can be processed into diameter 1-
1.5mm circular hole.The most narrow place of sample intake passage 8 is located at upper end, and sample holes 5 are gradually tapered up, seamlessly transitted to sample intake passage 8 most
Narrow place, then sample intake passage 8 gradually amplify, the lower end of sample intake passage 8 is connected with Embryo Culture chamber 2.The lower end connection of sample output passage 9
Embryo Culture chamber 2, sample output passage 9 is gradually tapered up, and the most narrow place channel sized of sample output passage 9 is 1.5-2.0mm, sample output passage 9
Most narrow place after again gradually amplify, connected using smooth arc transition with embryo's accommodating chamber 7.Because sample intake passage 8 is most narrow
Place is not more than 1.5mm, and the most narrow place channel sized of sample output passage 9 is 1.5-2.0mm, plays the speed for slowing down both sides fluid communication
Degree, is conducive to culture chamber environment to keep stable effect, in sample-adding sampling process, to the environment shadow of internal Embryo Culture chamber 2
Sound is very small, and the inside of the Embryo Culture unit is filled with advance using the good culture medium of carbon dioxide balance (it is recommended that using a journey
Formula culture medium), sealed by silica gel plug, in use, first then taking out sample introduction end plug hole in 37 DEG C of insulating box rewarmings about 45 minutes
3 silica gel plug, is put into after embryonated egg, and silica gel plug is stoppered at once, and the Embryo Culture unit is put into constant incubator, by
Smart ovum slowly spreads to Embryo Culture chamber 2.Because it is transparent material, Embryo Culture can be taken out in any desired time
Unit observes embryonic development situation, and camera dynamically observation Embryo Culture process can be also installed in constant incubator, has been observed
Finish, put back in insulating box and continue to cultivate；Such as need to take out embryo, stand up Embryo Culture unit, you can embryo is flowed into embryo
Accommodating chamber 7, then recovers normal horizontally-arranged, opens sampling plug, can pipe taking-up embryo using embryo transfer suction pipe；If it find that
Further culture is still needed to, handstand Embryo Culture unit makes embryo be back in Embryo Culture chamber 2, then horizontally-arranged continuation back to normal
Culture, can also be transferred in another new Embryo Culture unit and continue to cultivate.
In order to prevent that embryo flows back into sample intake passage 8, sample intake passage 8 and Embryo Culture during rotation Embryo Culture unit
The junction of chamber 2 is connected by one section of transition arc, and the bending direction of transition arc is curved with the sample intake passage wall of the junction
Qu Fangxiang and Embryo Culture cavity wall bending direction are on the contrary, the transition arc should be with sample intake passage 8 and the phase of tire tire culture chamber 2 certainly
Cut, to ensure that embryonated egg smoothly flows into Embryo Culture chamber 2.Change a kind of form of presentation, the lower end passage boring ratio embryo of sample intake passage 8
The hole of culture chamber 2 is small, and the junction of Embryo Culture chamber 2 and sample intake passage 8 passes through rapid desufflation 10 transition of section, rapid desufflation section 10
Playing prevents embryonated egg or embryo to be back to sample intake passage 8.
In the present embodiment, the bottom of Embryo Culture chamber 2 is curved, it is ensured that in incubation, and embryonated egg or embryo collect in
Together, and the lowest part of Embryo Culture chamber 2 is rested on.
In the present embodiment, the bottom of embryo's accommodating chamber 7 is curved, and the minimum point of embryo's accommodating chamber 7 is located at thief hole 6
Underface, conveniently take embryo.
In the present embodiment, from sample holes 5 to sample intake passage 8, then to Embryo Culture chamber 2, then to sample output passage 9, then to embryo
The whole vias inner walls of tire accommodating chamber 7 are seamlessly transitted, without turning and burr, and corner uses tangent circular arc
Transition so that embryonated egg or embryonic cell slidably, are not tangled.
In the present embodiment, multiple tracks transverse direction fin, can use screw thread on the side wall of sample introduction end plug hole 3 and sampling end consent 4
Shape, or thread like fin, horizontal fin can extrude silica gel plug, play sealing effectiveness, it is ensured that air tight.
In the present embodiment, the aperture outer of sample holes 5 is higher than the base plane in sample introduction end plug hole 3 so that aperture outer can
To squeeze into silica gel plug bottom surface, it is ensured that sealing effectiveness.Similarly, the aperture outer of thief hole 6 is put down higher than the bottom of sampling end consent 4
Face.
In the present embodiment, a diameter of 10mm of sample introduction end plug hole 3 and sampling end consent 4.Sample holes 5 are diameter 4mm circles
Type, as long as sample holes 5 are easy to be put into embryonated egg, herein under the premise of, sample holes are 5 mouthfuls as far as possible small.Thief hole 6 is ellipse,
Because when taking embryo, probe tube is usually to tilt insertion embryo accommodating chamber 7, is observed for convenience in sampling, so handle takes
It is processed into ellipse in sample hole 6.Optimal, thief hole 6 is major axis 6mm, short axle 3mm ellipse.
In the present embodiment, the dimensions of culture unit main body 1 is long 55mm, width 25mm, high 35mm.
In the present embodiment, the surface of culture unit main body 1 sets breach, and breach is used to fix elastic band, and elastic band is fixed
Silica gel plug, prevents silica gel plug from loosening, further, and culture unit main body 1 upper surface sets the neck for placing silica gel plug, in order to
Fixed placement after silica gel plug is extracted, unrest is not mixed up or is dropped.
Foregoing Embryo Culture unit manufacturing process is：Make and the sample introduction end plug hole 3 inside culture unit main body 1, sample introduction
The consistent integral inside molding of hole 5, sample intake passage 8, Embryo Culture chamber 2, embryo's accommodating chamber 7, thief hole 6, the shape of sampling end consent 4,
Fixed integral inside molding, moulding by casting culture unit main body 1, then in erosion removal integral inside molding, so as to obtain Embryo Culture list
Member, can inject in advance with the good embryo culture medium of carbon dioxide balance into Embryo Culture chamber 2 before dispatching from the factory, then use silica gel plug
Seal sample introduction end plug hole 3 and sampling end consent 4.The perishable metal of material selection of the integral inside molding is preferred, using acid or its
Its chemicals attack.The material of unit main body 1 is cultivated using glass or other any easily shapings, and nontoxic to embryo
, and not by the material of acid corrosion.
Load exhausted with after the good formula culture medium (see the past patent of invention) of carbon dioxide balance, constituting one in advance
To closed In vitro culture system, embryonated egg ectogenesis can be supported to blastaea rank directly in 37 DEG C of common insulating boxs
Section.For embryonated egg, this system has completely disengaged from the use of CO2gas incubator, and observes embryo's in confined conditions
Development, not only Embryo Culture stability more preferably, quality it is higher outer, moreover it is possible to significantly save the space of Embryo Culture room, largely subtract
CO is lacked2Usage amount, incubation step is significantly simplified, so that considerably reducing working strength and reduces operational error
Probability, realizes the safety for both protecting embryo and operating personnel, improves operating efficiency, work quality again and economizes on resources
Purpose.
The Embryo Culture unit of the present invention, is added after embryonated egg, can intuitively observe the growth course of embryo, installs shooting
Head, can dynamically be observed, and using preceding without carbon dioxide balance, during culture and observation, internal physiological environment will not change,
Whole device small volume, light weight can be used largely.User keeping temperature need to only balance in constant incubator.
The present invention is described by way of example above, but the invention is not restricted to above-mentioned specific embodiment, it is all to be based on
Any change or modification that the present invention is done belong to the scope of protection of present invention.
Claims (10)
1. a kind of Embryo Culture unit, it is characterized in that：Culture unit main body including transparent material, cultivates the upper table of unit main body
Face opens up in a sample introduction end plug hole and a sampling end consent, sample introduction end plug hole and sampling end consent and fills in plug seal, enters
The bottom in sample end plug hole sets sample holes, and the bottom of sampling end consent sets thief hole, and the underface of thief hole sets embryo to hold
Receive chamber, Embryo Culture chamber is set in culture unit main body, Embryo Culture chamber passes through sample intake passage and connects sample holes, Embryo Culture chamber
Embryo's accommodating chamber is connected by sample output passage.
2. Embryo Culture unit according to claim 1, it is characterized in that：The most narrow place of sample intake passage is located at upper end, sample introduction
Hole is gradually tapered up, and is seamlessly transitted to the most narrow place of sample intake passage, and then sample intake passage gradually amplifies, sample intake passage lower end and embryo
Culture chamber is connected；The lower end connection Embryo Culture chamber of sample output passage, sample output passage is gradually tapered up, after the most narrow place of sample output passage
Gradually amplify again, connected using smooth arc transition with embryo's accommodating chamber.
3. Embryo Culture unit according to claim 2, it is characterized in that：The junction of Embryo Culture chamber and sample intake passage leads to
Cross quick contraction section transition.
4. Embryo Culture unit according to claim 2, it is characterized in that：The narrowest diameter of sample intake passage is not more than
1.5mm, most narrow place's channel diameter size of sample output passage is 1.5-2.0mm.
5. Embryo Culture unit according to claim 1, it is characterized in that：Unit main body is cultivated using nontoxic to embryo saturating
Bright material, plug uses silica gel plug.
6. Embryo Culture unit according to claim 1, it is characterized in that：The bottom of Embryo Culture chamber is curved, and embryo holds
Receive chamber bottom it is curved, and embryo's accommodating chamber minimum point be located at thief hole underface.
7. Embryo Culture unit according to claim 1, it is characterized in that：On the side wall of sample introduction end plug hole and sampling end consent
Multiple tracks transverse direction fin, the aperture outer of sample holes is higher than the base plane in sample introduction end plug hole, and the aperture outer of thief hole, which is higher than, to be taken
The base plane in sample end plug hole.
8. Embryo Culture unit according to claim 1, it is characterized in that：Sample holes are diameter 4mm circular holes, and thief hole is
Major axis 6mm, short axle 3mm elliptical aperture.
9. a kind of preparation method of the Embryo Culture unit described in claim 1, it is characterized in that：Make first and culture unit master
The sample introduction end plug hole in internal portion, sample holes, sample intake passage, Embryo Culture chamber, embryo's accommodating chamber, thief hole, sampling end consent shape
The consistent integral inside molding of shape, fixed integral inside molding, moulding by casting culture unit main body, then erosion removal integral inside molding again, from
And obtain Embryo Culture unit.
10. the preparation method of Embryo Culture unit according to claim 9, it is characterized in that：Injected to Embryo Culture intracavitary
In advance with the good embryo culture medium of carbon dioxide balance, then sample introduction end plug hole and sampling end consent are sealed with silica gel plug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710390586.0A CN107022484A (en) | 2017-05-27 | 2017-05-27 | A kind of Embryo Culture unit and preparation method thereof |
Applications Claiming Priority (1)
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| US6193647B1 (en) * | 1999-04-08 | 2001-02-27 | The Board Of Trustees Of The University Of Illinois | Microfluidic embryo and/or oocyte handling device and method |
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