Acipenser dabryanus oosperm tissue slicing method

A technique for tissue sectioning and sturgeon, applied in the field of microscopic tissue sectioning, can solve the problems of low success rate of sectioning, inability to completely remove membranes, and narrowness, and achieves reduction of tedious operation steps, rich yolk material, and tough and dense egg membranes. Effect

Active Publication Date: 2016-05-11
FISHERIES INST SICHUAN ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem solved by the present invention is that the egg diameter of Sturgeon dabryi's eggs is relatively large, and the egg membrane is tough and dense. Eggs are covered with a thicker layer of mucous membrane, and the process of dehydration, transparency, and wax immersion is not as good as that of other fishes. Generally, the method of slicing fertilized eggs of fish is not suitable for sturgeons, and the success rate of slicing is relatively low. Low

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  • Acipenser dabryanus oosperm tissue slicing method

Examples

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Embodiment 1

[0031] A method for slicing tissues at the cleavage stage of fertilized eggs of Dabry's sturgeon, comprising the following steps:

[0032] Material fixation: Put the eggs of Sturgeon dabryi's that have developed to the cleavage stage of fertilized eggs into 50 times the volume of Bouin's fixative solution for 24 hours, wash them repeatedly with 70% ethanol, and then store them in 70% ethanol every other day Replace the 70% alcohol once for three consecutive times to fully wash off the fixative and save it for future use.

[0033] Dehydration: Gradient dehydration of fertilized eggs stored in 70% ethanol, 80% ethanol dehydration once for 2 hours, 90% ethanol dehydration once for 2 hours, 95% ethanol dehydration twice for 2 hours each time, and 100% ethanol dehydration twice, Each dehydration 1h.

[0034] Transparent: Put the dehydrated fertilized eggs into the transparent transition solution with a volume fraction of terpineol: absolute ethanol = 1:1 for 1 hour, then treat wit...

Embodiment 2

[0040] A method for slicing gastrula stage tissue of the fertilized eggs of Dabry's sturgeon, comprising the following steps:

[0041] Material fixation: fertilized eggs of Acipenser dabryi at different stages were fixed in 10% formaldehyde fixative solution for 24 hours, washed repeatedly with 70% ethanol, and then stored in 70% ethanol every other day Replace the 70% alcohol once for three consecutive times to fully wash off the fixative and save it for future use.

[0042] Dehydration: Gradient dehydration of fertilized eggs stored in 70% ethanol, 80% ethanol dehydration once for 2 hours, 90% ethanol dehydration once for 2 hours, 95% ethanol dehydration twice for 2 hours each time, and 100% ethanol dehydration twice, Each dehydration 1h.

[0043] Transparent: Put the dehydrated fertilized eggs into the transparent transition solution with volume fraction terpineol: absolute ethanol = 1:1 for 2 hours, then treat with terpineol for the first time for 4 hours, and for the sec...

Embodiment 3

[0049] A method for tissue slicing at the fertilized egg stage of Dabry's sturgeon zygote, comprising the following steps:

[0050] Material fixation: fertilized eggs of Acipenser dabryi at different stages were fixed in 50 times the volume of Bouin's fixative solution for 24 hours, washed repeatedly with 70% ethanol, and then stored in 70% ethanol, replaced every other day 70% alcohol, three times in a row to fully wash off the fixative, and save for future use.

[0051] Dehydration: Gradient dehydration of fertilized eggs stored in 70% ethanol, 80% ethanol dehydration once for 2 hours, 90% ethanol dehydration once for 2 hours, 95% ethanol dehydration twice, each time for 1 hour, and then 100% ethanol dehydration twice, Each dehydration 1h.

[0052] Transparent: Put the dehydrated fertilized eggs into the transparent transition solution with a volume fraction of terpineol: absolute ethanol = 1:1 for 2 hours, and then treat them with terpineol for 5 hours for the first time a...

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Abstract

The present invention relates to a microstructure slicing technology, and specifically relates to an acipenser dabryanus oosperm tissue slicing method. The acipenser dabryanus oosperm tissue slicing method mainly comprises seven steps of material fixation, dehydration, transparentizing, wax dipping, embedding, slicing and staining; a fixing liquid used in the material fixation is a Bouin's fixing liquid and a formaldehyde fixing liquid with the mass fraction of 10%; and the total dehydration time is 10-20h. The acipenser dabryanus oosperm tissue slicing method aims at sturgeon oosperm which belongs to a viscid egg, has the characteristics of being rich in vitellogenin, tough and dense in egg envelope, narrow in perivitelline space, and the like, a complete serial tissue slicing method suitable for sturgeon oosperm is established, the sturgeon oosperm can be efficiently processed, and better paraffin sections can be made.

Description

technical field [0001] The invention relates to a microscopic tissue slicing technique, in particular to a tissue slicing method for fertilized eggs of Dabry's sturgeon. Background technique [0002] Dabry's sturgeon (Acipenserdabryanus), also known as Yangtze River sturgeon and Shalazi, is a unique freshwater resident species of Acipenseridae (Acipenseridae) in my country. It is distributed in the upper reaches of the Yangtze River and the lower reaches of the Jinsha River (Ding Ruihua 1994). Animals and first-level urgently protected fish in the upper reaches of the Yangtze River (Liu Jun 2004), the International Union for Conservation Union (IUCN) critically endangered (critically endangered, CR) species (Du Jun, etc. 2009), the Convention on International Trade in Endangered Species of Wild Fauna and Flora , CITES) Appendix II protected species, one of the main protection objects of the Rare and Endemic Fish National Nature Reserve in the upper reaches of the Yangtze River...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/06
CPCG01N1/06G01N1/28
Inventor 刘亚赵柳兰杨淞龚全陈叶雨肖青杜军符红梅
Owner FISHERIES INST SICHUAN ACADEMY OF AGRI SCI
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