Method for simultaneously detecting completeness of sperm nucleus protein and membrane

A technique for integrity, nucleoproteins, applied in the experimental field

Inactive Publication Date: 2015-02-18
SHANDONG PROVINCIAL FAMILY PLANNING INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It solves the defects of a single experimental method, can evaluate sperm quality more accurately, and provides a method for simultaneously detecting sperm nucleoprotein and membrane integrity for clinical use

Method used

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  • Method for simultaneously detecting completeness of sperm nucleus protein and membrane
  • Method for simultaneously detecting completeness of sperm nucleus protein and membrane
  • Method for simultaneously detecting completeness of sperm nucleus protein and membrane

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Experimental program
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Embodiment Construction

[0041] The method for simultaneously detecting sperm nucleoprotein and membrane integrity, the steps are as follows:

[0042] ① Preparation of hypotonic solution

[0043] Hypotonic solution is a solution containing 0.735g sodium citrate and 1.35g fructose per 100ml of distilled water, with an ionic strength of 0.15 and an osmotic pressure of 150mOsm / kgH2O (mosmole, ie milliosm). The unit of the ionic strength is mol / kg.

[0044] ② Preparation of sperm morphology fixative

[0045] (1) Wash the yams, slice them, dry them at 54°C for 2 hours, and crush them to 90 mesh to obtain dried yams powder;

[0046] (2) Add the dried yam powder obtained in step (1) into petroleum ether to obtain a mixed slurry, reflux the mixed slurry at 90° C. for 45 minutes, and filter the mixed slurry to obtain a yam powder filter cake; the dried yam powder and petroleum The mass volume ratio of ether is 1g: 2~3ml;

[0047] (3) Dry the yam powder filter cake obtained in step (2) at 70° C. for 1.5 hou...

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Abstract

The invention relates to a method for simultaneously detecting completeness of a sperm nucleus protein and a membrane, which comprises the following steps: (1) preparing a hypotonic solution; (2) preparing a sperm morphology fixing solution; (3) carrying out simple sperm hypotension and morphology fixing treatment; (4) centrifuging the semen with fixed sperm morphology, standing to stratify, discarding the supernatant, and diluting the obtained semen with a phosphate buffer solution for sperm sediment to obtain a diluted semen; (5) sufficiently and evenly mixing the diluted semen and phosphate buffer solution, centrifugating, discarding the supernatant, washing the sperm precipitate with the phosphate buffer solution, and adding gelatin to the washed sperm precipitate to prepare a semen; (6) putting the prepared semen on a glass slide, and drying; (7) fixing the glass slide at 25 DEG C by adding the sperm morphology fixing solution; (8) adding aniline blue into the glass slide for dyeing; (9) flushing the glass slide with tap water, and immersing the glass slide in 0.9 wt% sodium chloride for decolorization; and (10) naturally airing or blow-drying the glass slide, adding 2 drops of glycerol sealing solution, and sealing with a cover slip.

Description

technical field [0001] The invention relates to an experimental method, specifically a method for simultaneously detecting sperm nucleoprotein and membrane integrity. Background technique: [0002] Cells carry out material exchange and metabolic activities through the cell membrane. Sperm are easily affected by external factors and chemical substances in the process of spermatogenesis. The ability of sperm membrane to transmit these substances directly affects the vitality and metabolism of sperm, which is related to whether sperm can be capacious and complete the whole process of fertilization. The nuclear maturation of human sperm is a key link in the structural and functional maturation of sperm, which is of great significance for maintaining the integrity of genetic material and ensuring the correct transmission of paternal genetic information. Impaired sperm nuclear maturation can have a greater adverse effect on sperm function and fertilization outcomes. The study f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
Inventor 张美华王磊光仕治达邱毅
Owner SHANDONG PROVINCIAL FAMILY PLANNING INST OF SCI & TECH
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