Cytology division method of scophthalmus maximus spermiogenesis phase

A technology of sperm cells and cytology, applied in the direction of scientific instruments, analytical materials, preparation of test samples, etc., can solve the problems of lack of system division, difficulty in observing and distinguishing the structure of sperm cells in different metamorphosis stages, limited, etc., and achieve high Reference and research value, improvement of dyeing uniformity, effect of enhancing image clarity

Active Publication Date: 2018-06-19
ZHEJIANG OCEAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] 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

Method used

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  • Cytology division method of scophthalmus maximus spermiogenesis phase

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Effect test

Embodiment 1

[0029] The cytological division method of turbot sperm metamorphosis stage comprises the following steps:

[0030]1) Collect the testis of turbot in stage IV-V, put the testis tissue in 2.0% glutaraldehyde solution at 1°C, dissolve 25% glutaraldehyde in phosphate buffer solution to obtain glutaraldehyde solution, Fix at 4°C for 3 hours; rinse with phosphate buffer for 3 times, and post-fix with 1.1% osmic acid with pH 7.18 at 4°C for 1.0 hour; 2) Wash the fixed testis tissue with phosphate buffer for 3 The second time, take 0.4% uranyl acetate solution, soak and stain the testis tissue at 1°C overnight, and take it out with CO2-free 2 Wash off the excess dye solution in distilled water, then rinse with phosphate buffer three times for use; 3) Dehydrate with ethanol solutions with gradient concentrations of 50%, 70%, 80%, 90% and 95%, and treat each concentration for 15 minutes , and then treated with 100% alcohol for 20 minutes, then treated with pure ethylene oxide for 20 mi...

Embodiment 2

[0040] The cytological division method of the metamorphosis stage of turbot sperm cells includes: fixation, pre-staining, dehydration and embedding, uranyl acetate staining, lead citrate staining, and slice analysis, specifically including the following steps:

[0041] 1) Fixation: Collect turbot testis in stage IV-V, place testis tissue in 2.5% glutaraldehyde solution at 4°C, dissolve 25% glutaraldehyde in phosphate buffer solution to obtain glutaraldehyde solution, fixed at 5°C for 4 hours; washed with phosphate buffer for 4 times, 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 substances in the 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 turn various components in the cells such as proteins, fats, carbohydrates, etc...

Embodiment 3

[0057] The cytological division method of the metamorphosis stage of turbot sperm cells includes: fixation, pre-staining, dehydration and embedding, uranyl acetate staining, lead citrate staining, and slice analysis, specifically including the following steps:

[0058] Fixation: Collect turbot testis in stage IV-V, place testis tissue in 2.5% glutaraldehyde solution at 2°C, dissolve 25% glutaraldehyde solution in phosphate buffer solution to obtain 2.5% glutaraldehyde solution Dialdehyde solution, fixed at 4°C for 3.5 hours; rinsed with phosphate buffer three times, and then post-fixed with 1.2% osmic acid at 4°C for 1.0 hour; fixation with glutaraldehyde solution and osmic acid can make sperm cells The substance is as close as possible to its morphological structure and position in its living state, which can prevent the autolysis and corruption of sperm cells, prevent the enzymes in the cells from decomposing proteins, and make various components in the cells such as proteins...

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Abstract

The invention discloses a cytology division method of a scophthalmus maximus spermiogenesis phase. The method comprises the following steps: fixing, pre-dyeing, dehydrating and embedding, dyeing by uranyl acetate, dyeing by lead citrate, and slicing preparing and analysis. A scophthalmus maximus spermiogenesis process is divided into 8 phases: comprising sperm cell I-VII stage and mature sperm. The method has the beneficial effects that the cytology division method overcomes the problem that during common histological slice observation, sperm cells at different metamorphosis periods cannot bedistinguished due to limited magnification times, pre-dyeing is helpful for increasing dyeing uniformity, tissue structure definition, and tissue dyeing contrast grade, dyeing effect can be optimized,and image definition of the tissue structure of the sperm cells can be obviously enhanced; the cytology division method can fill the research blank of phase division for the flounder paralichthys spermiogenesis, and can be used for cytology division of the spermiogenesis phase of the other seawater fish.

Description

technical field [0001] The invention relates to the field of fish sperm cell metamorphosis, in particular to a method for cytological division of turbot sperm cell metamorphosis stages. technical background [0002] Spermiogenesis, also known as spermatogenesis, is the final stage of spermatogenesis: the spermatid, which has completed the second meiosis, undergoes the condensation of the nucleus, the formation of the tail and the absorption of the residual body, and is formed by round cells transforms into mature spermatozoa (spermatozoa). The process of sperm cell metamorphosis determines the final structure assembly and activation function of sperm. This process directly determines the timing, quantity and quality of sperm production. [0003] 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 d...

Claims

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Application Information

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IPC IPC(8): G01N23/22G01N1/28G01N1/30G01N1/36
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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