CN107653188A - Seminal fluid adding set during a kind of Human Sperm egg's external fertilization - Google Patents

Seminal fluid adding set during a kind of Human Sperm egg's external fertilization Download PDF

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Publication number
CN107653188A
CN107653188A CN201710873065.0A CN201710873065A CN107653188A CN 107653188 A CN107653188 A CN 107653188A CN 201710873065 A CN201710873065 A CN 201710873065A CN 107653188 A CN107653188 A CN 107653188A
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CN
China
Prior art keywords
seminal fluid
needle
tip
outer tube
set during
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Pending
Application number
CN201710873065.0A
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Chinese (zh)
Inventor
饶金鹏
金敏
金帆
田申
钱小红
邱枫
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Zhejiang University ZJU
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Zhejiang University ZJU
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Publication date
Application filed by Zhejiang University ZJU filed Critical Zhejiang University ZJU
Priority to CN201710873065.0A priority Critical patent/CN107653188A/en
Publication of CN107653188A publication Critical patent/CN107653188A/en
Pending legal-status Critical Current

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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
    • C12M23/06—Tubular
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • C12M33/07—Dosage or metering devices therefore
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/44—Means for regulation, monitoring, measurement or control, e.g. flow regulation of volume or liquid level

Abstract

Seminal fluid adding set during a kind of Human Sperm egg's external fertilization, including seminal fluid suction sample needle, connection latex leather hose and sample-adding controller.Seminal fluid suction sample needle is the commercialization standard 3ml Pasteur glass pipettes that tip marks 30 μ l, 50 μ l, 80 μ l scales with emery wheel, and controller is 1ml standard injector for medical purpose.The present invention is simple in construction, it is handy to facilitate, the use of Pasteur glass pipette is compared to the special elongated plastic pipette tips of import, the plastics release harmful substance such as bisphenol-A is avoided while cost is greatly reduced potentially to damage to caused by sperm, it also avoid the long pipette tips of whole box because the opening time is long and existing for potential pollution risk.The connection of the syringe of latex leather hose and removal syringe needle uses, and because adding buffer field, compared to the controllability that General Purpose Rubber suction nozzle then improves operation, when being operated to micro liquid, effect is particularly evident.

Description

Seminal fluid adding set during a kind of Human Sperm egg's external fertilization
Technical field
The present invention relates to medical treatment, laboratory technique field, and seminal fluid adds during more particularly to a kind of Human Sperm egg's external fertilization Feeder apparatus.
Background technology
With the relieving of national " two children " policy comprehensively, increasing women (being greatly advanced age) has fertility side The demand in face.But with advancing age and current environment pollution, job and life stress be big and bad life style etc. is all Multifactor influence, there are about 1 pair in the current every 7 couples of Mr. and Mrs in China can not carry out normal reproduction, and this will be by supplementary reproduction (examination Pipe baby) technology.
And the realization of tube-test baby techniques, it is important that a link be embryo experiments room IVF-ET (in vitro fertilization-embryos Tire transplant) operation.And operating needs to carry out under the extremely strict environment of purity requirements, including laboratory integrated environment Cleanliness factor, the vessel contacted with gamete, embryo, the cleanliness factor etc. of reagent.
Operation in vitro fertilization is taken out simultaneously out of the wife's side body on the day of the sperm on the bridegroom's or husband's side is added to after the screening of centrifugation upstream The process of the corolla mound compound of in vitro culture for a period of time, it is the step of key one in whole Test-Tube Baby Lab operation.Mesh Before, it is then added in ovum nutrient solution and is completed using the special elongated plastic pipette tips absorption seminal fluid of 100 μ l pipettors installation import more This process.
But there are some researches show plastic products often containing the unfavorable healthy material such as bisphenol-A, have to gamete or embryo in the recent period Tire causes the risk of latent lesion.And special import spear head is generally 100-150 branch/whole box of box and purchased, cost is higher, and needs Using finishing in the term of validity behind Kaifeng, but even in the effect phase, the pipette tips behind Kaifeng still have by contaminated risk, and one Denier must abandon beyond effect phase, remaining long pipette tips, cause to waste.
The content of the invention
In view of the shortcomings of the prior art, the invention provides it is a kind of it is simple in construction, flexibly it is controllable, cost is cheap, and can have Effect avoids potentially toxic harmful substance from discharging, seminal fluid addition dress when evading the Human Sperm egg's external fertilization for depositing contaminated risk Put.
Technical proposal that the invention solves the above-mentioned problems is：
Seminal fluid adding set during a kind of Human Sperm egg's external fertilization, including suction sample needle 1 and sample-adding controller 3, and institute The rear end 12 for stating suction sample needle 1 is connected with the outlet 31 of the sample-adding controller 3 by latex leather hose 2；
The suction sample needle 1 of hard includes the needle body of tubulose, the rear end of the needle body and the one of the latex leather hose 2 End is connected, and the front end of the needle body is coaxially connected with tip 11, the tubulose that the tip 11 penetrates in two, and the tip 11 internal diameter is less than the internal diameter of the needle body, and scale 13 is provided with vertically on the outer wall of the tip 11, and tip 11 is transparent, just In observation；
The controller 3 is syringe, and the syringe includes outer tube and the core bar being slidably disposed in outer tube, core The front end of bar is located in outer tube and is provided with piston, and the rear end of core bar through the rear end of the outer tube and is provided with rear lug 32, and described The outlet 31 of outer tube front end is connected with the other end of the latex leather hose 2.
Further, the suction sample needle 1 is Pasteur glass pipette.
Further, the seminal fluid suction sample needle 1 marks 30 μ l, 50 μ l, 80 μ l scales 13 with emery wheel for tip 11 and identified 3ml Pasteur glass pipettes.
Seminal fluid suction sample needle of the present invention uses commercialization standard 3ml Pasteur glass pipettes, and glassware is compared to modeling Material, the harmful substance such as plastics release bisphenol-A causes potentially to damage to sperm when effectively can avoid aspirating or being loaded.Pasteur glass Glass suction pipe is the most frequently used consumptive material in embryo experiments room, it is instant still, occur in the absence of the long pipette tips of such as tailormadepiston because dosage is few Behind Kaifeng the problem of resting period long vulnerable to pollution.And the cost of single branch Pasteur glass pipette is only the 1/ of the long pipette tips of import 10-1/20, greatly reduce cost.
When carrying out operation in vitro fertilization, the final concentration after upstream dilutes again should be centrifuged according to sperm to draw certain body Product is added, and ensures that the final concentration by seminal fluid for filling ovum is (1-5) * 105/ ml, if too low by seminal concentration, have , if by seminal fluid excessive concentration, egg mother cell polyspermy risk be present in smart ovum fertilization risk of failure.Therefore when micro- in high power Microscopic observation Makler counting plate for sperm, if having 7-9 sperm in average 10 lattices, show that sperm centrifugation upstream dilutes again Final concentration of (7-9) * 10 afterwards6/ ml, seminal fluid is added in the micropore for the 750 μ l for filling ovum after this adjustment of 30 μ l is aspirated. With this similarly, if sperm concentration is (4-6) * 10 after adjustment6/ ml, 50 μ l of suction carry out seminal fluid addition；If sperm concentration after adjustment For 3*106/ ml, 80 μ l of suction carry out seminal fluid addition.Therefore, with reference to cylinder volume formula V=π (d/2)2H, by commercialization mark Quasi- 3ml Pasteurs glass pipette tip internal diameter d is about 2mm, and the length for obtaining 30 μ l fluid columns is 10mm, and the length of 50 μ l fluid columns is 16mm, and the length of 80 μ l fluid columns is 25mm.The emery wheel after the sterilizing of alcolhol burner flame envelope can be used in the corresponding positions of glass pipette tip Put the delineation mark for carrying out 30 μ l, 50 μ l, 80 μ l.It must not be marked using marking pen, because oily matter can volatilize VOC in marking pen (VOC), damage easily is produced to sperm and ovum.
Connection latex leather hose can be according to the operating habit and comfort level of each experiment operator from entire bundle emulsion tube Suitable length is voluntarily intercepted, with the 1ml standards injector for medical purpose combined use for eliminating syringe needle, protective cap, compared to traditional rubber Glue suction nozzle, because adding buffer field, the suction too quickly to gamete is avoided, preferably micro liquid can be controlled.And After liquid has been aspirated, it is not necessary to which hand pinches rubber suction nozzle always, can decontrol syringe rear lug, and the seminal fluid of suction is moved from test tube It is added into the micropore for fill egg mother cell so that operation is more freely.
Beneficial effects of the present invention are mainly manifested in：
It is poisonous and harmful that 1. the use of standard 3ml Pasteur glass pipettes of the present invention avoids bisphenol-A etc. in the long pipette tips of plastics Material latent lesion to caused by sperm, evade contaminated risk caused by long-term storage behind Kaifeng, and reduce cost.
2. Pasteur glass tube tip carries out the mark on scale with emery wheel so that seminal fluid is added in the control of amount more Accurately, oily matter VOC release has then effectively been evaded without using marking pen.
3. the connection of latex leather hose and syringe outer tube uses, compared to traditional rubber suction nozzle, can be more freely to micro Liquid is manipulated.
Brief description of the drawings
Fig. 1 is the structural representation of the present invention.
Embodiment
Reference picture 1, seminal fluid adding set during a kind of Human Sperm egg's external fertilization, including suction sample needle 1 and sample-adding control Device 3, and the rear end 12 of the suction sample needle 1 is connected with the outlet 31 of the sample-adding controller 3 by latex leather hose 2；
The suction sample needle 1 of hard includes the needle body of tubulose, the rear end of the needle body and the one of the latex leather hose 2 End is connected, and the front end of the needle body is coaxially connected with tip 11, the tubulose that the tip 11 penetrates in two, and the tip 11 internal diameter is less than the internal diameter of the needle body, and scale 13 is provided with vertically on the outer wall of the tip 11, and tip 11 is transparent, just In observation；
The controller 3 is syringe, and the syringe includes outer tube and the core bar being slidably disposed in outer tube, core The front end of bar is located in outer tube and is provided with piston, and the rear end of core bar through the rear end of the outer tube and is provided with rear lug 32, and described The outlet 31 of outer tube front end is connected with the other end of the latex leather hose 2.
The suction sample needle 1 is Pasteur glass pipette.
Seminal fluid suction sample needle 1 is tip 11 mark that 30 μ l, 50 μ l, 80 μ l scales 13 identify with emery wheel 3ml bars This moral glass pipette.
The connection sponge leather length of tube is suitable (can be flexible according to the distance between each hand of operating personnel two and comfort level Cut leather hose length), internal diameter 5mm, external diameter 7mm, thickness 1mm.
The sample-adding controller is to eliminate syringe needle, protective cap, and the 1ml standards for only retaining core bar, piston and overcoat are medical Syringe.
The outlet 31 of the rear end 12 of the seminal fluid suction sample needle and the sample-adding controller is entered by the latex leather hose 2 of centre Row is tightly connected.
The seminal fluid suction sample needle 1 is the commercialization standard that tip 11 marks 30 μ l, 50 μ l, 80 μ l scales 13 with emery wheel 3ml Pasteur glass pipettes.It according to the internal diameter of commercialization standard 3ml Pasteur glass pipettes tip 11 is about 2mm to be, in conjunction with circle Columnar volume formula V=π (d/2)2H, the length for obtaining 30 μ l fluid columns are 10mm, and the length of 50 μ l fluid columns is 16mm, and 80 μ l The length of fluid column is 25mm, and makees corresponding marking.The emery wheel after the sterilizing of alcolhol burner flame envelope can be used in glass pipette tip 11 Relevant position carries out 30 μ l, 50 μ l, 80 μ l scale 13 and marked.And must not be marked using marking pen, because of oiliness thing in marking pen Matter can volatilize VOC (VOC), easily produce damage to sperm and ovum.
The connection can flexibly be cut out with the length of latex leather hose 2 according to the distance between each hand of operating personnel two and comfort level Cut.Internal diameter is 5mm, external diameter 7mm, thickness 1mm so that seminal fluid can be aspirated sample needle by the both ends of latex leather hose 2 just Rear end 12 and the sealed envelope connection of outlet 31 of the sample-adding controller 3.
The sample-adding controller 3 is to eliminate syringe needle, protective cap, and the 1ml standards for only retaining core bar, piston and overcoat are medical Syringe.When carrying out Human in vitro Fertilization operation, by centrifuging the seminal fluid that dilutes again of upstream, by being seen under high-power microscope Makler counting plate for sperm is examined, according to different observation results：There is 7-9 sperm/4-6 sperm/3 essence in 10 lattices Son, judge that corresponding seminal fluid centrifugation upstream dilutes concentration after processing again：(7-9)*106/ml；(4-6)*106/ml；3*106/ml。 Now, the rear lug 32 of sample-adding controller 3 end is first pulled back, reserves one section of air, can will during finally to add seminal fluid All liq is got in fertilization micropore in pipe.Then, by the seminal fluid in seminal fluid adding set during the Human Sperm egg's external fertilization Suction sample needle 1 is extend into equipped with the 5ml round tubes of seminal fluid after processing so that the tip 11 of seminal fluid suction sample needle 1 is immersed Under the seminal fluid liquid level of ttom of pipe.At the same time, pull is loaded the rear lug 32 of the end of controller 3 backward, and eyes then observe seminal fluid Whether the corresponding scale 13 of suction sample needle 1 tip 11 is had been raised to：30μl/50μl/80μl.When the essence for being drawn into respective volume After liquid, loosen the rear lug 32 of sample-adding controller 3 end, carefully seminal fluid suction sample needle 1 is taken out from round tube, then moves to and puts There is the micropore of egg mother cell, most seminal fluid suction sample needle 1 is stretched into fertilization micropore at last, promotes sample-adding controller 3 end forward Rear lug 32, whole seminal fluid in pipe are added into containing 750 μ l in by the micropore of seminal fluid, now, all seminal fluid in vitro fertilization additions Operation is completed, and is filled the final concentration by seminal fluid of ovum and ought be in 1*105/ml—5*105In this section of/ml.
Content described in this specification embodiment is only enumerating to the way of realization of inventive concept, protection of the invention Scope is not construed as being only limitted to the concrete form that embodiment is stated, protection scope of the present invention also includes art technology Personnel according to present inventive concept it is conceivable that equivalent technologies mean.

Claims (3)

  1. A kind of 1. seminal fluid adding set during Human Sperm egg's external fertilization, it is characterised in that：Including suction sample needle (1) and sample-adding control Device (3) processed, and the rear end (12) of the suction sample needle (1) and the outlet (31) of the sample-adding controller (3) pass through sponge leather Pipe (2) is connected；
    The suction sample needle (1) of hard includes the needle body of tubulose, the rear end of the needle body and the one of the latex leather hose (2) End is connected, and the front end of the needle body is coaxially connected with tip (11), and the tip (11) is in the tubulose of two insertion, and described The internal diameter of tip (11) is less than the internal diameter of the needle body, is provided with scale (13), and point on the outer wall of the tip (11) vertically Portion 11 is transparent；
    The controller (3) is syringe, and the syringe includes outer tube and the core bar being slidably disposed in outer tube, core bar Front end be located in outer tube and be provided with piston, the rear end of core bar runs through the rear end of the outer tube and is provided with rear lug (32), and described The outlet (31) of outer tube front end is connected with the other end of the latex leather hose (2).
  2. 2. seminal fluid adding set during a kind of Human Sperm egg's external fertilization as claimed in claim 1, it is characterised in that：The suction Sample needle (1) is Pasteur glass pipette.
  3. 3. seminal fluid adding set during a kind of Human Sperm egg's external fertilization as claimed in claim 2, it is characterised in that：The seminal fluid Suction sample needle (1) is the 3ml Pasteurs glass suction that tip (11) mark 30 μ l, 50 μ l, 80 μ l scales (13) mark with emery wheel Pipe.
CN201710873065.0A 2017-09-25 2017-09-25 Seminal fluid adding set during a kind of Human Sperm egg's external fertilization Pending CN107653188A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710873065.0A CN107653188A (en) 2017-09-25 2017-09-25 Seminal fluid adding set during a kind of Human Sperm egg's external fertilization

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710873065.0A CN107653188A (en) 2017-09-25 2017-09-25 Seminal fluid adding set during a kind of Human Sperm egg's external fertilization

Publications (1)

Publication Number Publication Date
CN107653188A true CN107653188A (en) 2018-02-02

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Family Applications (1)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN2438429Y (en) * 2000-09-07 2001-07-11 李清芹 Double-chamber semen transplanting device
CN201958985U (en) * 2011-01-07 2011-09-07 郝天羽 Disposable semen liquefaction self processing appliance
CN205528761U (en) * 2015-12-23 2016-08-31 河北省沧州中西医结合医院 Move egg apparatus
US20170157327A1 (en) * 2014-02-07 2017-06-08 Panace Co., Ltd. Injection apparatus and injection method using same
CN207330956U (en) * 2017-09-25 2018-05-08 浙江大学 Sperm adding set during a kind of Human Sperm egg's external fertilization

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN2438429Y (en) * 2000-09-07 2001-07-11 李清芹 Double-chamber semen transplanting device
CN201958985U (en) * 2011-01-07 2011-09-07 郝天羽 Disposable semen liquefaction self processing appliance
US20170157327A1 (en) * 2014-02-07 2017-06-08 Panace Co., Ltd. Injection apparatus and injection method using same
CN205528761U (en) * 2015-12-23 2016-08-31 河北省沧州中西医结合医院 Move egg apparatus
CN207330956U (en) * 2017-09-25 2018-05-08 浙江大学 Sperm adding set during a kind of Human Sperm egg's external fertilization

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