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Application of percoll to improving separation efficiency of sperm flow cytometer

A technology of flow cytometry and separation efficiency, which is applied in the application field of sperm flow cytometry separation efficiency, can solve problems such as the joint use of flow cytometry technology, improve sperm motility, improve separation efficiency, and improve utilization rate effect

Inactive Publication Date: 2019-04-02
INNER MONGOLIA SAIKEXING LIVESTOCK BREEDING & SEED IND BIOTECH RES INST CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The percoll method has been widely used in artificial insemination, in vitro fertilization, embryo transfer and other assisted reproductive technology fields, but there is no related research on its combined use with flow cytometry

Method used

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  • Application of percoll to improving separation efficiency of sperm flow cytometer
  • Application of percoll to improving separation efficiency of sperm flow cytometer
  • Application of percoll to improving separation efficiency of sperm flow cytometer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: An application of using percoll to improve the separation efficiency of sperm flow cytometry is used to improve the efficiency of sperm flow cytometry separation, which includes the following steps:

[0029] (1) Semen test: using the pseudo-vaginal method to collect semen, the semen is milky white with no peculiar smell, the viability under the microscope is 0.35, and the semen density is 460 million / ml;

[0030] (2) Take 1.2 billion sperm, gently put them into the sperm separation solution, and centrifuge at 800g for 30min;

[0031] (3) Suck off the supernatant, add 5ml of sperm diluent to the bottom sediment, mix gently, 1500rpm, 5min, suck off the supernatant;

[0032] (4) Add 5ml of sperm diluent, 1500rpm, 5min, suck off the supernatant, and use flow cytometry to separate the sperm after staining;

[0033] (5) Detection:

[0034]

[0035] (Note: The dead sperm rate on the machine - the dead sperm rate displayed on the flow cytometer during sperm s...

Embodiment 2

[0036] Embodiment 2: An application of using percoll to improve the separation efficiency of sperm flow cytometry is used to improve the efficiency of sperm flow cytometry separation, which includes the following steps:

[0037] (1) Semen test: Use the false vagina method to collect semen. The semen is milky white, no peculiar smell, the viability under the microscope is 0.6, and the semen density is 800 million / ml;

[0038] (2) Take 500 million sperm, gently put them into the sperm separation solution, and centrifuge at 800g for 30min;

[0039] (3) Suck off the supernatant, add 5ml of sperm diluent to the bottom sediment, mix gently, 1500rpm, 5min, suck off the supernatant;

[0040] (4) Add 5ml of sperm diluent, 1500rpm, 5min, suck off the supernatant, and use flow cytometry to separate the sperm after staining;

[0041] (5) Detection:

[0042]

[0043] (Note: The dead sperm rate on the machine--the dead sperm rate displayed on the flow cytometer during sperm separation...

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Abstract

The invention relates to an application of percoll to improving separation efficiency of a sperm flow cytometer, belongs to the technical field of animal breeding, and is used for improving the separation efficiency of sperm flow cells. The application disclosed by the invention has the advantages that by a percoll method, the vitality of sperms can be notably improved, dead sperms can be removed,and the separation efficiency of the sperm flow cells can be effectively improved through treated sperms.

Description

technical field [0001] The invention belongs to the technical field of animal reproduction, and in particular relates to the application of using percoll reagent (silica gel particle suspension treated with polyvinylpyrrolidone) to process semen to remove dead sperm and improve the separation efficiency of sperm flow cytometry. Background technique [0002] The purposeful selection of sperm - the separation of X and Y sperm is the most ideal way to achieve sex control. Gender control can not only increase the intensity of selection, speed up the breeding process, but also give full play to the maximum economic benefits of gender-affected traits. The advent of flow cytometry has made it possible to commercialize sex-controlled separation technology. At present, the flow cytometry separation technology has been greatly improved, but the separation speed still cannot fully meet the market demand, and it is also one of the main reasons for the high price of sex-controlled froze...

Claims

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Application Information

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IPC IPC(8): C12N5/076
CPCC12N5/061C12N5/0612C12N2509/00
Inventor 李喜和苏杰
Owner INNER MONGOLIA SAIKEXING LIVESTOCK BREEDING & SEED IND BIOTECH RES INST CO LTD
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