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Method for rapidly separating sperm head, sperm tail and normal motile sperm

A sperm head and sperm technology, which is applied in the analysis of materials, individual particle analysis, particle and sedimentation analysis, etc., can solve the problems of inability to separate viable sperm at the same time, time-consuming, complicated operation, etc., and achieves simple sorting method and sperm purity. and vitality enhancing effect

Active Publication Date: 2020-06-23
NANTONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, scholars have used the method of density gradient centrifugation to separate sperm heads and sperm tails, but this method is time-consuming and complicated to operate, and on the other hand, it cannot separate viable sperm at the same time
However, although the upstream method, percoll and other low-speed centrifugation methods can obtain viable sperm, they cannot split to obtain high-purity sperm heads and tails.

Method used

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  • Method for rapidly separating sperm head, sperm tail and normal motile sperm
  • Method for rapidly separating sperm head, sperm tail and normal motile sperm
  • Method for rapidly separating sperm head, sperm tail and normal motile sperm

Examples

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Effect test

Embodiment 1

[0022] 1. Processing of mouse sperm samples

[0023] The mice were dissected, and the clean epididymis and vas deferens were separated and placed in a 2 cm petri dish containing 2 mL of HS solution, cut into pieces, incubated at a constant temperature of 35 °C for 15 min, and vibrated or blown 3-5 times in the middle to allow the sperm to fully dissociate. Filter through a 70 μm cell sieve to remove cell clusters and other impurities. Wash once with HS, 800 g, 5 min. Sperm pellet was resuspended in 2 mL of HS, 400 μL of sperm sample was reserved, and the remaining 1.5 mL of sperm sample was sonicated under the conditions of 500 W, ultra 5 s, and rest 5 s, for a total of 5 min.

[0024] 2. Preparation of sperm head positive control sample

[0025] Take 200 μL of sperm samples from the reserved 400 μL, add 10 μL of 10% SDS, 5 μL of 20 mg / mL proteinase K, and treat at 37 °C for 5 min. Pipette 10 μL and place it on a glass slide, cover it with a cover glass and observe the sper...

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Abstract

The invention discloses a method for rapidly separating a sperm head, a sperm tail and a normal motile sperm, and belongs to the technical field of reproductive medicine. The sperm, the sperm head andthe sperm tail are sorted by adopting a flow cytometry according to differences of cell lateral scattering light-area (SSC-A), forward scattering light-area (FSC-A), lateral scattering light-width (SSC-W) and lateral scattering light-height (SSC-H) in a sperm sample. By adopting the method disclosed by the invention, the sperm head, the sperm tail and the normal motile sperm can be quickly separated, purity is high, and the method is simple and convenient.

Description

technical field [0001] The invention belongs to the technical field of reproductive medicine and relates to a method for rapidly separating sperm heads, sperm tails and normal energetic sperm. Background technique [0002] Abnormal spermatogenesis is the main cause of male infertility, and abnormal sperm production is a common consequence of abnormal spermatogenesis. There are also various types of abnormal sperm, such as broken head, broken tail, folded tail, round head, etc., and the pathological mechanism of this kind of disease is far from elucidated. It has been clinically mature to select a single normal sperm, use ICSI (Intracytoplasmic sperm injection) to achieve in vitro fertilization, and then make these patients obtain offspring through IVF. However, research in recent years has shown that this kind of offspring not produced through natural conception has great health risks. Therefore, research has clarified the causes and pathological mechanisms of teratospermia...

Claims

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Application Information

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IPC IPC(8): G01N15/14
CPCG01N15/14G01N15/1429G01N2015/1477
Inventor 张小宁曾旭辉周雯雯黄荣祖顾亚云陈添兴
Owner NANTONG UNIVERSITY
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