CN112980777A - Method for screening sperms during artificial insemination - Google Patents
Method for screening sperms during artificial insemination Download PDFInfo
- Publication number
- CN112980777A CN112980777A CN201911312212.2A CN201911312212A CN112980777A CN 112980777 A CN112980777 A CN 112980777A CN 201911312212 A CN201911312212 A CN 201911312212A CN 112980777 A CN112980777 A CN 112980777A
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- CN
- China
- Prior art keywords
- transparent material
- artificial insemination
- microscope
- ovum
- culture solution
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Links
- 230000009027 insemination Effects 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 14
- 238000012216 screening Methods 0.000 title abstract description 6
- 239000012780 transparent material Substances 0.000 claims abstract description 22
- 210000004681 ovum Anatomy 0.000 claims abstract description 15
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 13
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 13
- 210000004027 cell Anatomy 0.000 claims abstract 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 18
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 6
- 229930091371 Fructose Natural products 0.000 claims description 5
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 5
- 239000005715 Fructose Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 102000016943 Muramidase Human genes 0.000 claims description 3
- 108010014251 Muramidase Proteins 0.000 claims description 3
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 3
- 235000014655 lactic acid Nutrition 0.000 claims description 3
- 239000004310 lactic acid Substances 0.000 claims description 3
- 239000004325 lysozyme Substances 0.000 claims description 3
- 235000010335 lysozyme Nutrition 0.000 claims description 3
- 229960000274 lysozyme Drugs 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 238000013459 approach Methods 0.000 claims description 2
- 239000012531 culture fluid Substances 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 235000013772 propylene glycol Nutrition 0.000 claims 1
- 230000035935 pregnancy Effects 0.000 abstract 1
- 235000013601 eggs Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 208000000509 infertility Diseases 0.000 description 2
- 230000036512 infertility Effects 0.000 description 2
- 231100000535 infertility Toxicity 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 206010002659 Anovulatory cycle Diseases 0.000 description 1
- 206010003883 azoospermia Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 208000008634 oligospermia Diseases 0.000 description 1
- 230000036616 oligospermia Effects 0.000 description 1
- 231100000528 oligospermia Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0604—Whole embryos; Culture medium therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Developmental Biology & Embryology (AREA)
- Reproductive Health (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Gynecology & Obstetrics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A method of selecting sperm cells for artificial insemination comprising: the method comprises the following steps: and arranging a culture solution on the transparent material. Step two: and laying the ovum at the designated position, and laying the transparent material under a microscope. Step three: arranging sperms at the other end of the transparent material, and finishing after observation meets the requirement of artificial insemination. The method can increase the screening of sperms at least to a certain extent and can avoid the phenomenon of polyspermy at least to a certain extent. It is a feature of the present invention to vary the difficulty of insemination by extending the distance between the ovum and sperm and adjusting the consistency of the culture. The invention can also be used for replacing other artificial insemination schemes during pregnancy.
Description
Technical Field
The disclosure relates to the technical field of human assisted reproduction, in particular to a method for screening sperms in artificial insemination.
Background
The test tube infant is one of the main methods for treating infertility at present, female patients with ovulation failure or male patients with oligospermia and weak sperm can breed by using the test tube infant technology, so that the infertility caused by diseases is solved, but the test tube infant technology has limited screening of the sperm. It is a feature of the present invention to vary the difficulty of insemination by extending the distance between the ovum and sperm and adjusting the consistency of the culture.
Disclosure of Invention
According to one aspect of the present disclosure, there is provided a method of screening sperm during artificial insemination, comprising:
the method comprises the following steps: and arranging a culture solution on the transparent material.
Step two: and laying the ovum at the designated position, and laying the transparent material under a microscope.
Step three: arranging sperms at the other end of the transparent material, and finishing after observation meets the requirement of artificial insemination.
Preferably, the transparent material is a supporting material having a sufficient length to be placed under a microscope, such as a glass sheet or a transparent sheet (or transparent paddle) made of other material.
Preferably, the area of the transparent material for disposing the culture solution may be a straight line or a curved line, and may have a relatively complex shape such as a U-shape or an L-shape.
Preferably, the culture solution is a liquid for connecting the sperm and the ovum.
Preferably, as an alternative embodiment, the main components of the culture fluid can be water, fructose, propylene glycol, and a proper amount of lysozyme and lactic acid can be added to more closely approach the vaginal fluid components.
Further, the culture solution comprises the following main components in parts by weight: fructose (about 2 percent), propylene glycol (about 0.8 percent), and the balance of water, and the mixture ratio can be modified according to actual needs.
Preferably, the viscosity degree of the culture solution can be modified by modifying the content of the propylene glycol according to requirements.
Preferably, as an alternative embodiment, the main component of the culture solution can be water, fructose, propylene glycol (or the like), the ph value of the culture solution can be made between 3.5 and 6.5 by modifying the ratio of lactic acid, and lysozyme can be added in small amounts according to sperm quality or need.
Further, the culture solution comprises the following main components in parts by weight: fructose (about 2 percent), propylene glycol (about 0.8 percent), and the balance of water, and the proportion can be modified according to actual requirements.
Preferably, an egg is generally placed on one end of the transparent material, and then the transparent material is placed under a microscope to observe the egg.
Preferably, the objective and eyepiece are adjusted to the appropriate position, taking into account that the objective should avoid hitting the sample.
Preferably, a sufficient amount of sperm is disposed at the other end of the transparent material connected by the culture solution.
Preferably, the microscope is observed to determine that artificial insemination is substantially complete.
Alternatively, the microscope may be adjusted to meet the requirement of human eyes for observing the insemination of the ovum, for example, an alternative configuration is to select a 20-fold objective lens, combine with a 30-fold eyepiece (or a camera with a display with similar magnification), and when adjusting the combination of the objective lens and the eyepiece magnification, use the microscope ruler to see if the 0.01mm scale can be observed as expected.
Alternatively, to more clearly identify the distance to the egg placement point, a scale may be added at the edge of the sperm placement area to indicate the distance to the egg placement area.
Optionally, as an alternative implementation, a computer may be used to obtain image frames of a camera connected to the microscope to monitor the images, and if the image change of a certain continuous image frame is greater than a certain threshold, a sound is prompted or/and the screen is flashed to reduce the number of times that the person observes the initial stage.
Further, the image change of the consecutive picture frames means that a pixel change of the consecutive image frames or a change ratio of pixel-by-pixel comparison of two image frame information spaced by several frames satisfies more than a certain value.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
Fig. 1 schematically shows a layout, top view, of the transparent material according to the invention placed on top.
Fig. 2 schematically shows a layout, top view, of the transparent material according to the invention placed on top.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1
Step 1: the culture medium is added to the transparent sheet in a manner as shown in FIG. 1.
Step 2: eggs were added and placed under a microscope.
And step 3: adjusting the microscope to meet the requirement of observing the insemination condition of the ovum by human eyes, for example, an alternative configuration method is to select a 20-fold objective lens and combine a 30-fold eyepiece lens (or a camera with a similar magnification factor with a display), and of course, the objective lens and the eyepiece lens can also be adjusted according to the requirement and imaging, and when the objective lens and the eyepiece lens are adjusted in combination, a microscope ruler can be used to check whether the scale with 0.01mm can be observed according to the expected condition.
And 4, step 4: the observation (or observations) is continued until insemination is found to be complete.
Example 2
Step 1: the culture medium is added to the transparent sheet in a manner as shown in FIG. 1.
Step 2: adding ovum, placing under microscope, and using camera to replace eyepiece to transfer image frame information into computer.
And step 3: the focal length of the microscope and the objective lens used for adjusting the focal length of the microscope to meet the requirement of observing the insemination condition of the ovum in the image acquired by the computer, for example, an optional configuration method is to select the objective lens of 20 times, combine with the camera (camera or microscope camera) of 30 times, of course, the multiple of the objective lens can also be adjusted according to the requirement and imaging, and when the multiple of the objective lens is adjusted, a microscope ruler can be used for checking whether the scale of 0.01mm can be observed according to the expected condition.
And 4, step 4: at the beginning, the computer can be waited to send out a prompt to indicate that the sperm is present around the ovum.
And 5: the observation (or observations) is continued until insemination is found to be complete.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. A method of selecting sperm cells for artificial insemination comprising:
the method comprises the following steps: arranging a culture solution on the transparent material;
step two: laying ova at a designated position, and laying the transparent material under a microscope;
step three: arranging sperms at the other end of the transparent material, and finishing after observation meets the requirement of artificial insemination.
2. The method of claim 1, wherein the step of disposing the culture solution on the transparent material comprises:
the transparent material is a material with enough length and can be placed under a microscope for observation;
the culture solution is used for connecting sperms and ova;
as an alternative embodiment, the main components of the culture fluid can be water, fructose, propylene glycol, and lysozyme and lactic acid can be added in proper amount to more closely approach the components of vaginal fluid.
3. "step two" according to claim 1, wherein the placing of the ovum at the designated location and the placing of the transparent material under the microscope comprises:
generally, an ovum is arranged at one end of the transparent material, and then the transparent material is placed under a microscope to observe the ovum;
the objective and eyepiece are adjusted to the appropriate position taking into account that the objective should avoid hitting the sample.
4. "step three" according to claim 1, wherein sperm is arranged at the other end of the transparent material, and observation is completed after satisfying artificial insemination, comprising:
disposing a sufficient amount of sperm at the other end connected by the culture solution;
observations were made to determine that artificial insemination was substantially complete.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911312212.2A CN112980777A (en) | 2019-12-18 | 2019-12-18 | Method for screening sperms during artificial insemination |
| PCT/CN2020/134481 WO2021121077A1 (en) | 2019-12-18 | 2020-12-08 | Method for screening sperm during artificial insemination |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911312212.2A CN112980777A (en) | 2019-12-18 | 2019-12-18 | Method for screening sperms during artificial insemination |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112980777A true CN112980777A (en) | 2021-06-18 |
Family
ID=76344076
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911312212.2A Pending CN112980777A (en) | 2019-12-18 | 2019-12-18 | Method for screening sperms during artificial insemination |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN112980777A (en) |
| WO (1) | WO2021121077A1 (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8390681B1 (en) * | 2008-12-23 | 2013-03-05 | LifeCell Dx, Inc. | Computer assisted semen analyzer to analyze digital video clips received from a remote location |
| CN105861423A (en) * | 2016-04-19 | 2016-08-17 | 湖北省农业科学院畜牧兽医研究所 | Making method of operation vessel for preparation of pig ICSI zygotes |
| CN110074849A (en) * | 2019-05-09 | 2019-08-02 | 异起(上海)智能科技有限公司 | A kind of fertilization of natural imitation, in conjunction with artificial intelligence Insemination procedures and imitate the device of female reproduction environment |
-
2019
- 2019-12-18 CN CN201911312212.2A patent/CN112980777A/en active Pending
-
2020
- 2020-12-08 WO PCT/CN2020/134481 patent/WO2021121077A1/en active Application Filing
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| Publication number | Publication date |
|---|---|
| WO2021121077A1 (en) | 2021-06-24 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210618 |
|
| WD01 | Invention patent application deemed withdrawn after publication |