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Cytological division method of sperm cell metamorphosis stage in turbot

A sperm cell and cytology technology, applied in scientific instruments, analytical materials, preparation of samples for testing, etc., can solve the problems of lack of systematic division, difficulty in observing and distinguishing the structure of sperm cells in different metamorphosis stages, limited and other problems. Reference and research value, the effect of improving dyeing uniformity and enhancing image clarity

Active Publication Date: 2021-11-16
ZHEJIANG OCEAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Unfortunately, reports on the histology and submicroscopic structure of sperm cells during metamorphosis in fish are very limited and very scattered. Due to the limitation of the magnification of ordinary light microscopes, it is difficult to observe and distinguish sperm cells in different stages of metamorphosis by histological section method. cell structure
Therefore, there is still a lack of systematic division of the sperm metamorphosis stages of seawater fish including turbot, resulting in a stagnation in the study of sperm metamorphosis in seawater fish

Method used

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  • Cytological division method of sperm cell metamorphosis stage in turbot

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Effect test

Embodiment 1

[0030] The cytological division method of the metamorphosis stage of turbot sperm cells, including the following steps:

[0031]1) Collect the turbot testis in the IV-V stage, place the testis tissue in a 2.0% glutaraldehyde solution at 1°C, and dissolve 25% glutaraldehyde in a phosphate buffer solution to obtain a glutaraldehyde solution. Fixed at 4°C for 3 hours; rinsed with phosphate buffer for 3 times, and post-fixed with 1.1% osmic acid at pH 7.18 at 4°C for 1.0 hours; 2) Rinse the fixed testis tissue with phosphate buffer 3 Second, take 0.4% uranyl acetate solution to soak and stain the testis tissue overnight at 1°C, remove it with no CO 2 Wash off excess dye solution in distilled water, and then rinse with phosphate buffer for 3 times before use; 3) Dehydrate with ethanol solutions with gradient concentrations of 50%, 70%, 80%, 90% and 95%, and treat each concentration for 15 minutes , then treated with 100% alcohol for 20 minutes, then treated with pure ethylene oxid...

Embodiment 2

[0041] The cytological division method of the metamorphosis stage of turbot sperm cells, including: fixation, pre-staining, dehydration and embedding, uranyl acetate staining, lead citrate staining, and preparation analysis, including the following steps:

[0042] 1) Fixation: Collect turbot testis in stage IV-V, place the testis tissue in 4°C, 2.5% glutaraldehyde solution, and dissolve 25% glutaraldehyde in phosphate buffer solution to obtain glutaraldehyde solution, fixed at 5°C for 4 hours; rinsed 4 times with phosphate buffer, and post-fixed with 1.2% osmic acid at 5°C for 1.3 hours; fixation with glutaraldehyde solution and osmic acid can make the substance in sperm cells as much as possible The morphological structure and position when it is close to its living state can prevent the autolysis and corruption of sperm cells, prevent the enzymes in the cells from decomposing proteins, and convert various components in the cells such as proteins, fats, carbohydrates, etc. int...

Embodiment 3

[0058] The cytological division method of the metamorphosis stage of turbot sperm cells, including: fixation, pre-staining, dehydration and embedding, uranyl acetate staining, lead citrate staining, and preparation analysis, including the following steps:

[0059] Fixation: The testis of turbot at stage IV-V were collected, and the testis tissue was placed in 2.5% glutaraldehyde solution at 2°C, and 25% glutaraldehyde solution was dissolved in phosphate buffer solution to obtain 2.5% glutaraldehyde solution. Dialdehyde solution, fixed at 4°C for 3.5 hours; washed three times with phosphate buffer, and post-fixed with 1.2% osmic acid for 1.0 hours at 4°C; glutaraldehyde solution and osmic acid fixation can make sperm cells The morphological structure and position of the substance as close as possible to its living state can prevent the autolysis and corruption of sperm cells, prevent the enzymes in the cell from decomposing the protein, and convert various components in the cell...

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Abstract

The invention discloses a cytological division method of the metamorphosis stage of turbot sperm cells, including: fixation, pre-staining, dehydration and embedding, staining with uranyl acetate, staining with lead citrate, preparation and analysis of the sperm cells of turbot The metamorphosis process is divided into 8 stages: including sperm cell Ⅰ-Ⅶ stage and mature sperm. Beneficial effects are: this method of cytology division overcomes the problem of inability to distinguish sperm cells in different metamorphosis stages due to limited magnification in ordinary histological section observation, and pre-staining helps to improve the uniformity of staining, the clarity of tissue structure and the contrast of tissue staining The degree of staining can be optimized, and the image clarity of the sperm cell structure can be significantly enhanced; the cytological division method of the sperm metamorphosis stage of turbot not only fills the research gap of the sperm cell metamorphosis stage division of flounder fish, but also can be applied Cytological division of the metamorphic stages of sperm cells in other marine fishes.

Description

technical field [0001] The invention relates to the field of fish sperm cell metamorphosis, in particular to a cytological division method for the metamorphosis stage of turbot sperm cells. [0002] technical background [0003] Spermiogenesis, also known as spermatogenesis, is the final stage of spermatogenesis: sperm cells that complete the second meiosis (spermatid) undergo nucleus condensation, tail formation and remnant absorption, and are formed by round cells. Transforms into mature sperm (spermatozoa). The process of sperm cell metamorphosis determines the final structural assembly and activation of spermatozoa. This process directly determines the timing, quantity and quality of sperm production. [0004] Unfortunately, the reports on histology and submicroscopic structure of sperm cells during metamorphosis in fish are very limited and scattered. Due to the limitation of the magnification of ordinary optical microscopes, it is difficult to observe and distinguish ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N23/22G01N23/2202G01N1/28G01N1/30G01N1/36
CPCG01N1/28G01N1/2813G01N1/30G01N1/36G01N23/22G01N23/2202G01N2001/364
Inventor 柳意樊刘清华李军徐世宏王彦丰
Owner ZHEJIANG OCEAN UNIV
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