CN207313583U - A kind of sperm screening based on microflow control technique and bionic device in vitro fertilization - Google Patents
A kind of sperm screening based on microflow control technique and bionic device in vitro fertilization Download PDFInfo
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Abstract
The utility model provides a kind of sperm screening based on microflow control technique and bionic device in vitro fertilization, including bionic device body, the bionic device body is by slide layer, silica gel pipe layers and dimethyl silicone polymer layer composition, central pool is equipped with the middle part of the silica gel pipe layers, entrance pool is equipped with the outside of the central pool, first outlet pond and second outlet pond, the central pool and entrance pool, first outlet pond and second outlet pond pass through the first pipeline respectively, second pipe and the connection of the 3rd pipeline, first pipeline, the first adding mouth is further respectively had on second pipe and the 3rd pipeline, second adding mouth and the 3rd adding mouth.This bionic device simulates natural conception process completely, avoids potential for adverse effects of the various operations to gamete and embryo, and can avoid the generation of polyspermy；The embryo quality under simulation natural conception state can be obtained, success rate in vitro fertilization can be greatly improved, the potential cause for causing infertility can be identified, so as to point the direction for clinical treatment.
Description
Technical field
The utility model belongs to biological technical field, is related to a kind of micro fluidic device, more particularly to a kind of based on micro-fluidic
Technology sperm screening with bionic device in vitro fertilization and should based on microflow control technique sperm screen with it is in vitro fertilization bionical
The preparation method and method of work of device.
Background technology
Infertile incidence is about 15%~20% in world wide, and has cumulative year after year trend.It is pre- according to WHO
Survey, in the near future, healthy reproduction problem will be that the third-largest disease of human health is threatened after cardiovascular and cerebrovascular disease and tumour
Disease.The increase of infertile incidence, and the relieving of national " two children " policy, have promoted the hair of auxiliary procreation technology significantly
Exhibition.First generation tube-test baby techniques --- IVF-ET Technique (IVF-ET) is since 1978 carry out, to many
Infertility Mr. and Mrs bring Gospel；Second generation tube-test baby techniques --- intracytoplasmic sperm injection (ICSI) technology and
The further abundant and perfect auxiliary procreation technology of third-generation technology --- preimplantation genetic diagnosis (PGD).Nevertheless,
It is many during auxiliary procreation technology to operate the damage that cause embryo, embryo's agensis or miscarriage are ultimately resulted in, this
It is the reason for causing tube-test baby techniques success rate to be difficult to more than 50%.These operations for having damage include：After sperm swim-up
Centrifugally operated, density gradient centrifugation operation and separating liquid in some components influence, wear ovum operation and culture environment and culture
Influence of liquid component etc..And the influence that avoid these from operating, it is in vitro fertilization to simulate natural conception as far as possible.
The normal process of becoming pregnant is needed by three phases：The generation of sperm and maturation；Sexuality；Sperm by cervix,
Uterine cavity, fallopian tubal smoothly reach ampulla and meet with ovum.Wherein, first and phase III and infertile relation the most
Closely.Spermatogenesis with it is ripe abnormal it is most common show as oligospermatism and asthénospermie, but these patients always have part
The normal sperm of vigor, the technology of the common high-quality sperm of separation in laboratory is upper reaches method and density-gradient centrifugation method, these are grasped
High-quality sperm of opposing can cause to damage, and can reach Natural Selection vigor eupyrene sperm by micropore depth difference on micro-fluidic chip
Purpose；Running resistance of the sperm in female genital tract is heavy, as cervical mucus can to sperm motility rise screening effect, only
The stronger sperm of vigor could march on towards uterus through cervical mucus；Selective barrier is played in uterus and fallopian tube junction, can be with
Stop that weak sperm passes through；Oviductal Fluid is conducive to capacitation, prepares for next step acrosome reaction and sperm-egg fusion；And ovum is thin
The secretion of born of the same parents and cumulus cell has chemotaxis screening effect.Therefore in the case of natural conception, only with good vigor and
The sperm of chemotactic response ability could complete fertilization process, and microfluidic chip technology can screen such sperm, and in vitro
Simulate this natural conception process.
The content of the invention
Purpose of utility model：For overcome the deficiencies in the prior art, the purpose of this utility model is to provide a kind of based on micro-
Fluidics sperm screening with bionic device in vitro fertilization and should based on microflow control technique sperm screen with it is in vitro fertilization
The preparation method and method of work of bionic device, this device simulate natural conception process completely, avoid external centrifugation, washing,
Potential for adverse effects of the various operations such as ovum to gamete and embryo is worn, and the generation of polyspermy can be avoided.
Technical solution：A kind of sperm screening and bionic device in vitro fertilization, including bionic device based on microflow control technique
Body, the bionic device body is by slide layer, silica gel pipe layers (SU-8 layer of silica gel) and dimethyl silicone polymer (PDMS) layer
Composition, the silica gel pipe layers middle part are equipped with central pool, are equipped with entrance pool on the outside of the central pool, first outlet pond and second goes out
Mouth pond, the central pool pass through the first pipeline, second pipe and the 3rd respectively with entrance pool, first outlet pond and second outlet pond
Pipeline connects, and the first adding mouth, the second adding mouth and the are further respectively had on first pipeline, second pipe and the 3rd pipeline
Three adding mouths.Sperm screening and bionic device in vitro fertilization, structure setting described in the utility model based on microflow control technique
Compact and reasonable, simulates natural conception process completely, avoids and centrifuges, washs in vitro, wearing the various operations such as ovum to gamete and embryo
Potential for adverse effects, and the generation of polyspermy can be avoided.Simulation natural conception shape can be obtained using this utility model device
Embryo quality under state, can greatly improve success rate in vitro fertilization.Can be identified using this device causes the potential of infertility
Reason, such as sperm can not penetrate cervical mucus, capacitation and acrosome reaction obstacle occur, so as to point the direction for clinical treatment.
This utility model device is applicable not only to the reproductive center of for example each medical institutions of human reproduction, and is suitable for animal breeding.
Further, above-mentioned sperm screening and bionic device in vitro fertilization, the central pool based on microflow control technique
Add sperm nutrient solution；Semen sample is added in the entrance pool；The first outlet pond and second outlet pond add ovarian follicle training
Nutrient solution and/or ovarian cumulus compound；The filling cervical mucus of first pipeline；Second pipe and the 3rd filling Oviductal Fluid of pipeline.
It is rational in infrastructure, can be bionical with high-quality.
Further, above-mentioned sperm screening and bionic device in vitro fertilization, the central pool based on microflow control technique
Radius 5mm, depth 0.1mm；The entrance pool radius is 2mm, depth 6mm；The first outlet pond and second outlet pond half
Footpath is 4mm, depth 6mm；First pipeline, second pipe and the 3rd pipeline long 10mm, it is 50 μm wide, it is 20 μm high.Structure is closed
Reason, it is easy to accomplish.
Further, the above-mentioned sperm screening based on microflow control technique and bionic device in vitro fertilization, first pipe
Road, second pipe and the 3rd pipeline are centered on central pool, respectively in 120 ° of settings.It is reasonable to set, it is easy to accomplish.
Further, the above-mentioned sperm screening based on microflow control technique is glued with bionic device in vitro fertilization, the uterine neck
Liquid is from male spouse, jenny or is artificial cervical mucus, and the artificial cervical mucus includes egg white；The sperm
Nutrient solution includes commercialization Earle ' s liquid, BWW liquid；The Oviductal Fluid includes commercialization HTF liquid, MHTF liquid；The follicle culture liquid
Including commercial KSOM nutrient solutions, CZB nutrient solutions；The ovarian cumulus compound is derived from male with occasionally jenny.Various nutrient solutions
It is wide with ovarian cumulus compound source, using convenient.
Further, the above-mentioned sperm screening based on microflow control technique and bionic device in vitro fertilization, the sperm training
Nutrient solution, it is composed of the following components：NaCl 0.721g, MgSO47H2O 0.247g, fructose 1g, Na2HPO4 3.581g,
KH2PO4 0.136g, pure water 1000ml.Sperm nutrient solution component is reasonable, and the vigor of sperm can be effectively ensured, and effect is good.
The utility model also provides a kind of sperm screening and the preparation of bionic device in vitro fertilization based on microflow control technique
Method, comprises the following steps：
(1) the film type mask plate of silica gel pipe layers is made；
(2) light transmission features of film are utilized, the silica gel pipe layers are obtained by photoresist developing photoetching；
(3) dimethyl silicone polymer prepolymer and curing agent are pressed 10:1 mixing, is sufficiently stirred, is inhaled bubble with vacuum pump
Go out, then slowly pour in silica gel pipe layers, vacuum outgas, 1.5~2h curing moldings in 72 DEG C of baking ovens, obtain the poly- diformazan
Radical siloxane layer；
(4) after cured dimethyl silicone polymer is cut, with card punch, centre is beaten above central pool and entrance pool
The hole of diameter 4mm, centre beats the hole of diameter 8mm above first outlet pond and second outlet pond, in the first adding mouth, second
The hole of diameter 2mm is beaten at adding mouth and the 3rd adding mouth；
(5) after being cleaned with ultrasonic cleaning machine, cleaned with deionized water, dried up with nitrogen；
(6) dimethyl silicone polymer layer is bonded with slide layer with plasma generator, obtains and described be based on microflow control technique
Sperm screening with bionic device in vitro fertilization.
Sperm screening and the preparation method of bionic device in vitro fertilization described in the utility model based on microflow control technique,
Method is reasonable, and application is convenient, is adapted to large-scale industrial production, and production cost is low.
The utility model also provides a kind of sperm screening and the work of bionic device in vitro fertilization based on microflow control technique
Method, comprises the following steps：
(1) after using the first pipeline of deionized water rinsing, second pipe and the 3rd pipeline, dried in 60 DEG C of baking ovens；
(2) use sample injector from the filling cervical mucus of the first adding mouth into the first pipeline, filling total amount is 10nl；
(3) managed respectively with sample injector from the second adding mouth and the 3rd filling Oviductal Fluid of adding mouth to second pipe and the 3rd
In road, filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool；
(5) add follicle culture liquid in first outlet pond and second outlet pond and ovarian cumulus compound, follicle culture liquid add
It is 350 μ l to enter amount；
(6) 100 μ l of semen liquefaction sample are added in entrance pool, with diaphragm seal capping bionic device surface, that is, are covered
Central pool, entrance pool, first outlet pond, second outlet pond, the first adding mouth, the second adding mouth and the 3rd adding mouth, then will
Bionic device body is horizontally placed in 37 DEG C of wet box；
(7) observed per 10min under inverted microscope once, monitor sperm operating condition；Sperm reaches first outlet pond
Continue 2~3d of culture with second outlet pond and with after ovum fertilization, taking out embryonated egg, obtain fertilization embryo.
Sperm screening and the method for work of bionic device in vitro fertilization described in the utility model based on microflow control technique,
Method is reasonable, and using that conveniently, can obtain fertilization embryo in vitro, the fertilization radicle of acquisition matches somebody with somebody occasionally female uterus according to male
Fertilization embryonic implantation is entered male in due course and matched somebody with somebody occasionally in female uterus by developmental state.
Further, above-mentioned sperm screening and the method for work of bionic device in vitro fertilization based on microflow control technique,
Comprise the following steps：
(1) after using the first pipeline of deionized water rinsing, second pipe and the 3rd pipeline, dried in 60 DEG C of baking ovens；
(2) use sample injector from the filling cervical mucus of the first adding mouth into the first pipeline, filling total amount is 10nl；
(3) managed respectively with sample injector from the second adding mouth and the 3rd filling Oviductal Fluid of adding mouth to second pipe and the 3rd
In road, filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool；
(5) sperm nutrient solution is added in first outlet pond and second outlet pond, addition is 350 μ l；
(6) 100 μ l liquefied semen samples are added in entrance pool, with diaphragm seal capping bionic device surface, that is, are sealed
Lid central pool, entrance pool, first outlet pond, second outlet pond, the first adding mouth, the second adding mouth and the 3rd adding mouth, then
Bionic device body is horizontally placed in 37 DEG C of wet box；
(7) sperm cultures of the 5 μ l containing sperm is taken after 50min-120min from first outlet pond and/or second outlet pond
Liquid, and carry out sperm count.
The application of this bionic device is convenient, and sperm can be screened, and obtains the sperm that form is normal, vigor is good.
Further, above-mentioned sperm screening and the method for work of bionic device in vitro fertilization based on microflow control technique,
The semen sample in entrance pool is suctioned out after step (7), 100 new μ l semen samples is rejoined, repeats the above steps, can be with
Achieve the purpose that to be enriched with motile sperm.
Further, above-mentioned sperm screening and the method for work of bionic device in vitro fertilization based on microflow control technique,
Comprise the following steps：
(1) after using the first pipeline of deionized water rinsing, second pipe and the 3rd pipeline, dried in 60 DEG C of baking ovens；
(2) use sample injector from the filling cervical mucus of the first adding mouth into the first pipeline, filling total amount is 10nl；
(3) managed respectively with sample injector from the second adding mouth and the 3rd filling Oviductal Fluid of adding mouth to second pipe and the 3rd
In road, filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool；
(5) Oviductal Fluid is added in first outlet pond and second outlet pond, addition is 350 μ l, the Oviductal Fluid
In contain an ovarian cumulus compound；
(6) 100 μ l liquefied semen samples are added in entrance pool, with diaphragm seal capping bionic device surface, that is, are sealed
Lid central pool, entrance pool, first outlet pond, second outlet pond, the first adding mouth, the second adding mouth and the 3rd adding mouth, then
Bionic device body is horizontally placed at 37 DEG C, is cultivated in 5%CO2 incubators；
(7) observe in first outlet pond and/or second outlet pond whether there is sperm with inverted microscope after 1h, seen after 4~6h
Examine whether there is second polar body discharge or female-male pronucleus formed；If second polar body discharge or female-male pronucleus formed, prompt sperm with
Ovum has been fertilized to form embryonated egg, at this time can suction out embryonated egg, and transplanting is supplied after continuing culture；If no second polar body discharge
Or female-male pronucleus is formed, then continue to observe 2h, until sperm and Folliculogenesis embryonated egg.
This bionic device method is reasonable, can be applied to sperm in vitro fertilization-embryonic implantation (IVF-ET), compared to facing at present
Bed routine IVF-ET, pregnancy rate can increase by 15%~20%, and polyspermy rate and abortion ratio can significantly reduce.
Above-mentioned technical proposal can be seen that compared with prior art, and the utility model has the advantages that：
(1) the utility model device simulates natural conception process completely, avoids body during existing auxiliary procreation technology
It is outer to centrifuge, wash, wearing potential for adverse effects of the various operations such as ovum to gamete and embryo, and the generation of polyspermy can be avoided.
(2) the utility model device can obtain the embryo quality under simulation natural conception state, can greatly improve external
The success rate of fertilization.
(3) the utility model device can identify the potential cause for causing infertility, as sperm can not penetrate cervical mucus, obtain
And acrosome reaction obstacle etc. can occur, so as to point the direction for clinical treatment.
(4) the utility model device is applied widely, this device can be used in human reproduction and animal breeding.
Brief description of the drawings
Fig. 1 is the structure diagram of silica gel pipe layers described in the utility model；
Fig. 2 is the sperm screening described in the utility model based on microflow control technique and bionic device section view in vitro fertilization
Structure diagram；
In figure：100 bionic device bodies, 1 central pool, 2 entrance pools, 3 first outlet ponds, 4 second outlet ponds, 5 first pipes
Road, 6 second pipes, 7 the 3rd pipelines, 8 first adding mouths, 9 second adding mouths, 10 the 3rd adding mouths.
Embodiment
Below in conjunction with the accompanying drawings and specific embodiment, the utility model is furtherd elucidate.
Embodiment 1
The screening of the sperm based on microflow control technique as shown in Figs. 1-2 and bionic device in vitro fertilization, including bionic device
Body 100, the bionic device body 100 is by slide layer, silica gel pipe layers (SU-8 layer of silica gel) and dimethyl silicone polymer
(PDMS) layer forms, and central pool 1 is equipped with the middle part of the silica gel pipe layers, and the outside of central pool 1 goes out equipped with entrance pool 2, first
Mouth pond 3 and second outlet pond 4, the central pool 1 pass through first respectively with entrance pool 2, first outlet pond 3 and second outlet pond 4
Pipeline 5, second pipe 6 are connected with the 3rd pipeline 7, are further respectively had on first pipeline 5,6 and the 3rd pipeline 7 of second pipe
First adding mouth 8, the second adding mouth 9 and the 3rd adding mouth 10.
Wherein, the central pool 1 adds sperm nutrient solution；Semen sample is added in the entrance pool 2；The first outlet
Pond 3 and second outlet pond 4 add follicle culture liquid and/or ovarian cumulus compound；First pipeline, the 5 filling cervical mucus；Second
6 and the 3rd pipeline of pipeline, 7 filling Oviductal Fluid.
As shown in appendix 1, the 1 radius 5mm of central pool, depth 0.1mm；2 radius of entrance pool is 2mm, and depth is
6mm；4 radius of the first outlet pond 3 and second outlet pond is 4mm, depth 6mm；First pipeline 5,6 and of second pipe
3rd pipeline 7 long 10mm, it is 50 μm wide, it is 20 μm high.Also, first pipeline 5,6 and the 3rd pipeline 7 of second pipe are with central pool
Centered on 1, respectively in 120 ° of settings.It is reasonable to set, it is easy to accomplish.
The volume parameter of the difference little Chi of subordinate list 1 and pipe layers
Embodiment 2
In the screening of the sperm based on microflow control technique of embodiment 1 and bionic device in vitro fertilization, the liquid in each little Chi
Source is as follows：
Semen sample can derive from the mankind or other animals；
The cervical mucus is from male spouse, jenny or is artificial cervical mucus, the artificial cervical mucus
Including egg white；The sperm nutrient solution includes commercialization Earle ' s liquid, BWW liquid；The Oviductal Fluid includes HTF liquid, MHTF
Liquid；The follicle culture liquid includes commercialization KSOM nutrient solutions, CZB nutrient solutions；The ovarian cumulus compound is derived from male with occasionally female
Property animal, from ampulla of uterine tube obtain.
Embodiment 3
Artificial cervical mucus, can take egg white with physiological saline 1:2~1:5 dilutions.
The sperm nutrient solution can be prepared voluntarily, composed of the following components：NaCl 0.721g, MgSO47H2O
0.247g, fructose 1g, Na2HPO4 3.581g, KH2PO4 0.136g, pure water 1000ml.
Embodiment 4
Sperm screening based on microflow control technique and the preparation method of bionic device in vitro fertilization, bag in a kind of embodiment 1
Include following steps：
(1) with reference to the accompanying drawings 1, attached drawing 2 and 1 content of subordinate list draw design drawing, and with specific software such as AutoCAD design chips
Pipeline and raised structures domain, and change into printable film format file；Silica gel pipe layers are made according to design drawing
Film type mask plate；
(2) light transmission features of film are utilized, the silica gel pipe layers (silicon substrate SU-8 moulds are obtained by photoresist developing photoetching
Tool, the thickness of SU-8 can be adjusted)；
(3) dimethyl silicone polymer prepolymer and curing agent are pressed 10:1 mixing, is sufficiently stirred, is inhaled bubble with vacuum pump
Go out, then slowly pour in silica gel pipe layers, vacuum outgas, 1.5~2h curing moldings in 72 DEG C of baking ovens, obtain the poly- diformazan
Radical siloxane layer；
(4) by cured dimethyl silicone polymer cut after, with card punch above central pool 1 and entrance pool 2 centre
The hole of diameter 4mm is beaten, centre beats the hole of diameter 8mm above first outlet pond 3 and second outlet pond 4, in the first adding mouth
8th, the hole of diameter 2mm is beaten at the second adding mouth 9 and the 3rd adding mouth 10；
(5) after being cleaned with ultrasonic cleaning machine, cleaned with deionized water, dried up with nitrogen；
(6) dimethyl silicone polymer layer is bonded with slide layer with plasma generator, obtains and described be based on microflow control technique
Sperm screening with bionic device in vitro fertilization.
Embodiment 5
A kind of sperm screening and the method for work of bionic device in vitro fertilization based on microflow control technique, including following step
Suddenly：
(1) with after the first pipeline of deionized water rinsing 5,6 and the 3rd pipeline 7 of second pipe, dried in 60 DEG C of baking ovens；
(2) use single pushing-type sample injector (available commercial) from 8 filling cervical mucus of the first adding mouth into the first pipeline 5,
Filling total amount is 10nl；
(3) respectively with single pushing-type sample injector from the second adding mouth 9 and 10 filling Oviductal Fluid of the 3rd adding mouth to the second pipe
In 6 and the 3rd pipeline 7 of road, filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool 1；
(5) follicle culture liquid and ovarian cumulus compound, follicle culture liquid are added in first outlet pond 3 and second outlet pond 4
Addition is 350 μ l；
(6) 100 μ l of semen liquefaction sample are added in entrance pool 2, with diaphragm seal capping bionic device surface, that is, are covered
Central pool 1, entrance pool 2, first outlet pond 3, second outlet pond 4, the first adding mouth 8, the second adding mouth 9 and the 3rd adding mouth
10, then bionic device body 100 is horizontally placed in 37 DEG C of wet box；
(7) observed per 10min under inverted microscope once, monitor sperm operating condition；Sperm reaches first outlet pond 3
Continue 2~3d of culture with second outlet pond 4 and with after ovum fertilization, taking out embryonated egg, obtain fertilization embryo.
Fertilization embryonic implantation is entered male in due course with occasionally female uterus developmental state according to male to move with occasionally female
Thing intrauterine.
Embodiment 6
The above-mentioned screening of the sperm based on microflow control technique is used for the work side of sperm screening with bionic device in vitro fertilization
Method, comprises the following steps：
(1) with after the first pipeline of deionized water rinsing 5,6 and the 3rd pipeline 7 of second pipe, dried in 60 DEG C of baking ovens；
(2) use sample injector from 8 filling cervical mucus of the first adding mouth into the first pipeline 5, filling total amount is 10nl；
(3) respectively with sample injector from the second adding mouth 9 and 10 filling Oviductal Fluid of the 3rd adding mouth to second pipe 6 and the
In three pipelines 7, filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool 1；
(5) Oviductal Fluid is added in first outlet pond 3 and second outlet pond 4；
(6) 100 μ l liquefied semen samples are added in entrance pool 2, with diaphragm seal capping bionic device surface, that is, are sealed
Lid central pool 1, entrance pool 2, first outlet pond 3, second outlet pond 4, the first adding mouth 8, the second adding mouth 9 and the 3rd adding mouth
10, then bionic device body 100 is horizontally placed in 37 DEG C of wet box；
(7) after 30min in the sperm nutrient solution of central pool visible motile sperm, as time went on, motile sperm by
More than cumulative and enter the microchannel of the liquid containing HTF, movable essence can be seen after about 50min in first outlet pond 3 and second outlet pond 4
Son, takes the HTF liquid containing sperm in 5 μ l outlet baths after 2h and carries out sperm count, if sperm sum up to 50000, can be direct
The sperm row screened with this is (IVF) in vitro fertilization, the sperm of this quantity enough with 1 oocyte fertilization.
By the screening of the present apparatus, sperm motility is more than 90%, and normal morphology sperm percentage also significantly improves, remote high
In 4%.If the number of motile sperm obtained is less, the semen sample in entrance pool can be suctioned out, rejoin 100 new μ l
Semen sample, so as to achieve the purpose that to be enriched with motile sperm.
Embodiment 7
The above-mentioned screening of the sperm based on microflow control technique is applied to sperm in vitro fertilization-embryo with bionic device in vitro fertilization
The method of work of (IVF-ET) is transplanted, is comprised the following steps：
(1) with after the first pipeline of deionized water rinsing 5,6 and the 3rd pipeline 7 of second pipe, dried in 60 DEG C of baking ovens；
(2) use sample injector from 8 filling cervical mucus of the first adding mouth into the first pipeline 5, filling total amount is 10nl；
(3) respectively with sample injector from the second adding mouth 9 and 10 filling Oviductal Fluid (HTF) of the 3rd adding mouth to second pipe
6 and the 3rd in pipeline 7, and filling total amount is 10nl；
(4) 10 μ l of sperm nutrient solution are added in central pool 1；
(5) Oviductal Fluid is added in first outlet pond 3 and second outlet pond 4, addition is 350 μ l, the fallopian tubal
Contain an ovarian cumulus compound in liquid；
(6) 100 μ l liquefied semen samples are added in entrance pool 2, with diaphragm seal capping bionic device surface, that is, are sealed
Lid central pool 1, entrance pool 2, first outlet pond 3, second outlet pond 4, the first adding mouth 8, the second adding mouth 9 and the 3rd adding mouth
10, bionic device body 100 is then horizontally placed at 37 DEG C, is cultivated in 5%CO2 incubators；
(7) observe sperm is whether there is in first outlet pond (3) and/or second outlet pond (4) with inverted microscope after 1h, 4~
Observation whether there is second polar body discharge after 6h or female-male pronucleus is formed；If second polar body discharge or female-male pronucleus are formed, prompt
Sperm has been fertilized to form embryonated egg with ovum, at this time can suction out embryonated egg, and transplanting is supplied after continuing culture；If without the second pole
Body is discharged or female-male pronucleus is formed, then continues to observe 2h, until sperm and Folliculogenesis embryonated egg.Using this device compared to mesh
Preceding routine clinical IVF-ET, pregnancy rate can increase by 15%~20%, and polyspermy rate and abortion ratio can significantly reduce.
The above is only the preferred embodiment of the utility model, it is noted that for the common skill of the art
For art personnel, on the premise of the utility model principle is not departed from, to central pool 1, entrance pool 2, first outlet pond 3, second
Outlet bath 4, the first pipeline 5,6 and the 3rd pipeline 7 of second pipe, the first adding mouth 8, the second adding mouth 9 and the 3rd adding mouth 10
Shape and size can be improved, these improvement are regarded as the scope of protection of the utility model.
Claims (3)
1. a kind of sperm screening based on microflow control technique and bionic device in vitro fertilization, it is characterised in that：Including bionic device
Body (100), the bionic device body (100) is made of slide layer, silica gel pipe layers and dimethyl silicone polymer layer, described
Be equipped with central pool (1) in the middle part of silica gel pipe layers, equipped with entrance pool (2), first outlet pond (3) and the on the outside of the central pool (1)
Two outlet baths (4), the central pool (1) pass through first respectively with entrance pool (2), first outlet pond (3) and second outlet pond (4)
Pipeline (5), second pipe (6) are connected with the 3rd pipeline (7), first pipeline (5), second pipe (6) and the 3rd pipeline (7)
On further respectively have the first adding mouth (8), the second adding mouth (9) and the 3rd adding mouth (10).
2. the sperm screening according to claim 1 based on microflow control technique exists with bionic device in vitro fertilization, its feature
In：Central pool (1) the radius 5mm, depth 0.1mm；Entrance pool (2) radius is 2mm, depth 6mm；Described first goes out
Mouth pond (3) and second outlet pond (4) radius is 4mm, depth 6mm；First pipeline (5), second pipe (6) and the 3rd pipe
Road (7) long 10mm, it is 50 μm wide, it is 20 μm high.
3. sperm screening and bionic device in vitro fertilization, its feature according to claim 1 or 2 based on microflow control technique
It is：First pipeline (5), second pipe (6) and the 3rd pipeline (7) are set in 120 ° respectively centered on central pool (1)
Put.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721309032.5U CN207313583U (en) | 2017-10-12 | 2017-10-12 | A kind of sperm screening based on microflow control technique and bionic device in vitro fertilization |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721309032.5U CN207313583U (en) | 2017-10-12 | 2017-10-12 | A kind of sperm screening based on microflow control technique and bionic device in vitro fertilization |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN207313583U true CN207313583U (en) | 2018-05-04 |
Family
ID=62429856
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| CN201721309032.5U Active CN207313583U (en) | 2017-10-12 | 2017-10-12 | A kind of sperm screening based on microflow control technique and bionic device in vitro fertilization |
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| Country | Link |
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2017
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