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FFRC strain common carp mature ovarian tissue frozen section making method

A technique for ovarian tissue and frozen section, which is applied in the preparation of test samples, sampling devices, etc., can solve the problems of localization research of unfavorable genes, and the pretreatment method of frozen section degreasing has not been reported, and achieves clear structure and shortening. effect of cycles

Inactive Publication Date: 2016-10-26
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

[0003] With the development and maturity of the gonads of Furui carp, the accumulation of fat gradually increases, which has a great impact on the dehydration process of ovarian tissue frozen sections and the observation of the tissue structure after sectioning, and is not conducive to the later use of immunohistochemistry for gene positioning research.
At present, many studies are aimed at the improved method of gene localization process such as immunohistochemistry, and the method of pre-fat removal for frozen sections has not been reported yet.

Method used

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  • FFRC strain common carp mature ovarian tissue frozen section making method
  • FFRC strain common carp mature ovarian tissue frozen section making method

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Experimental program
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Embodiment

[0018] 1. Prepare the required reagents before sampling

[0019] (1) 1×PBS:

[0020] ① Weigh the following reagents and place them in a 1L beaker: NaCl: 8g; KCl: 0.2g; Na2HPO4·12H2O: 3.58g; KH2PO4: 0.27g;

[0021] ② Add about 800ml of deionized water to the beaker, stir well to dissolve;

[0022] ③ Add NaOH to adjust the pH to 7.4, then add deionized water to make the volume to 1L;

[0023] ④ After high temperature and high pressure sterilization, store at room temperature.

[0024] (2) 4% paraformaldehyde: 4g paraformaldehyde + 100mL 1×PBS, add a little NaOH, heat in a water bath at 60°C until completely dissolved. Store in a refrigerator at 4°C, with a shelf life of 1 week. Ready to use.

[0025] (3) Sucrose solution:

[0026] 10% sucrose solution: 10g sucrose + 90ml 1×PBS;

[0027] 20% sucrose solution: 20g sucrose + 80ml 1×PBS;

[0028] 30% sucrose solution: 30g sucrose + 70ml 1×PBS;

[0029] 2. Frozen section preparation

[0030] (1) Fixation: The ovarian tissue...

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Abstract

The invention relates to a FFRC strain common carp mature ovarian tissue frozen section making method and belongs to the technical field of tissue sections. Mature ovarian tissues are taken out from a FFRC strain common carp, and a FFRC strain common carp mature ovarian tissue frozen section is obtained after fixation, degreasing, dehydration, embedding, freezing and slicing are performed. Compared with a common general freezing and slicing method, degreasing is performed through pure acetone, excessive fat in a later development period of FFRC strain common carp mature ovaries can be efficiently removed, and accordingly the frozen ovarian section high in quality and clearer in structure is obtained. An important and reliable molecular experiment material is provided for tissue localization and functional study such as immunohistochemistry of FFRC strain common carp mature ovarian genes, and the FFRC strain common carp mature ovarian tissue frozen section making method has the important significance on molecular mechanism study of good economic characters of FFRC strain common carps.

Description

technical field [0001] The invention relates to a method for making frozen slices of mature ovarian tissue of Furui carp, belonging to the technical field of tissue slices. Background technique [0002] Furui carp (FFRC Cyprinus carpio) is an aquatic product obtained by using Jian carp and Yellow River carp as the original parents, using the method of complete diallel hybridization, and with the assistance of PIT markers, through one-generation population selection and four consecutive generations of BLUP family selection. Breeding new species has achieved great genetic progress and significantly improved growth characteristics. It is a new breed of bulk freshwater aquaculture fish mainly promoted by the Ministry of Agriculture during the "Twelfth Five-Year Plan" and has been promoted in most areas of the country. With the development of molecular biology, it will become the development trend of aquatic breeding technology to explore the molecular mechanism of the excellent ...

Claims

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Application Information

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IPC IPC(8): G01N1/06G01N1/36G01N1/42
CPCG01N1/06G01N1/36G01N1/42
Inventor 王兰梅董在杰
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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