Seawater fish ovum electron-microscope scanning sample preparing method

A sample preparation and egg technology, which is applied in the field of seawater fish egg sample preparation, can solve the problems of egg brittleness, collapse, and difficulty in electron microscope observation, and achieve the effect of slow and stable dehydration, maintaining morphological characteristics, and maintaining egg quality

A sample preparation and egg technology, which is applied in the field of seawater fish egg sample preparation, can solve the problems of egg brittleness, collapse, and difficulty in electron microscope observation, and achieve the effect of slow and stable dehydration, maintaining morphological characteristics, and maintaining egg quality

CN100427918CInactive Publication Date: 2008-10-22SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Embodiment Construction

[0019] The present invention will be further described in detail below by taking an electron microscope scanning sample prepared from the ovum test material of the representative seawater famous and high-quality fish, the flat bream, as an example.

[0020] Oocyte source: when the diameter of oocytes of flat sea bream oocytes is greater than 400 μm in vivo, the luteinizing hormone releasing hormone analogue (LHRH-A 2 ) and chorionic gonadotropin (HCG) mixed hormones to induce ripening and ovulation. Injected in two divided doses for a total dose of 7.5 μg LHRH-A 2 +1500IU HCG / KgBW, adjust the water temperature to 21-22°C, reduce the light intensity, promote the continuous development and maturation of oocytes, and finally break away from the ovarian follicle membrane and enter the ovarian cavity, ovulate smoothly, and provide materials for the preparation of egg samples.

[0021] The sample preparation steps for egg electron microscope scanning are as follows:

[0022] 1. Pr...

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Abstract

The disclosed preparation method for ovum of sea fish as TEM sample comprises: fixing fresh fish ovum by glutaral solution diluted by phosphate buffer, cleaning, dewatering the ovum by 20-70% alcohol with concentration gradient increased by 10% and 75-100% alcohol with concentration gradient increased by 5% as two stages, then dewatering in pure acetone with CaCl2 as agent; then, drying at critic point, and spraying gold for the sample. This invention has ovum completion rate as 93.8-96.7% with low cost and convenient operation.

Description

technical field [0001] The invention relates to a preparation technology of seawater fish egg samples for electron microscope scanning observation. Background technique [0002] Fish fertilization includes two types of in vitro fertilization and internal fertilization. The fertilization process is that the sperm pass through the membrane hole of the egg and fuse with the egg nucleus to complete the combination of the gametes of the two sexes. Since the sperm of teleosts generally do not have acrosomes, the combination of mammalian sperm and eggs is due to the fact that fertilin and antifertilin recognize and interact with each other and produce sperm acrosomes in different fertilization methods. There is a sunken, usually funnel-shaped egg membrane pore on the shell membrane, which provides the only channel for the combination of sperm and eggs in teleosts, making the fertilization process have remarkable specificity (Wang Ruixia, 1984; Kuchnow, 1987; Kobayashi, 1987). How ...

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Application Information

Patent Timeline
22 Oct 2008
Publication
CN100427918C
IPC
G01N1/28; G01N13/10; G01Q30/20
Inventors
蔡泽平; 胡超群