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Preparation method of skin microscopic tissue slice of hynobiidae animals

An animal skin and microstructure technology, which is applied to the preparation of amphibian skin microtissue slices and the preparation of salamander skin tissue slices, can solve the problem of brittle and hard tissue blocks during wax immersion and poor dehydration effect of n-butanol. In order to achieve the effect of complete sectioning, improving transparency, and preventing incomplete penetration

Inactive Publication Date: 2016-01-13
HENAN UNIV OF SCI & TECH
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  • Abstract
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  • Claims
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Problems solved by technology

However, this method has the following deficiencies: (1) incomplete dehydration, resulting in incomplete paraffin penetration, resulting in broken tissue or cavities during sectioning, disintegration of sections during spreading, poor support and coloration during staining, etc.; ( 2) The dehydration effect of n-butanol is not good; (3) Due to the lack of a transitional step in the wax immersion process, the xylene carried from the transparent step enters the paraffin, resulting in a decrease in the paraffin concentration. In addition, the wax immersion time is too long to cause the tissue block to be brittle and hard

Method used

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  • Preparation method of skin microscopic tissue slice of hynobiidae animals

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Embodiment 1

[0029] Preparation of staining solution and differentiation solution:

[0030] Hematoxylin dye solution: Dissolve 1g hematoxylin in 20mL 100% ethanol to obtain liquid A, and 50g potassium alum in 300mL distilled water to obtain liquid B; mix liquid A and liquid B, boil for 2~3min, and add distilled water to fix Volume up to 1000mL; add 0.2g sodium iodate at the end to get.

[0031] Eosin dye solution: add 0.5 g of eosin to 100 mL of 85% ethanol and dissolve it completely. If it is not dyed, it can be diluted with 0.2% glacial acetic acid to promote eosin dyeing.

[0032] HCl-ethanol differentiation solution: add 1 mL of hydrochloric acid to 99 mL of 70% ethanol and mix well to obtain.

[0033] The preparation method of the skin microscopic tissue section of the small salamander (Hynobiuschinesis) in this embodiment includes the following steps:

[0034] (1) Take the material: at room temperature, use tweezers to pick up the skin on the back of the small salamander, then use scissors t...

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Abstract

The invention discloses a preparation method of a skin microscopic tissue slice of hynobiidae animals and belongs to the technical field of microscopic tissue slices. The preparation method includes following steps: 1) dehydration: successively soaking skin tissue, which has been subjected to fixing treatment, of the hynobiidae animals into ethanol solutions being 50%, 70%, 80%, 90%, 95% and 100% in concentration gradient to dehydrate the skin tissue, wherein dehydration time in each concentration gradient is 45-60 min; (2) transparent treatment: soaking the dehydrated skin tissue into a mixed liquid of 100% ethanol and dimethylbenzene for 2-3 min, and then soaking the dehydrated skin tissue into dimethylbenzene for 10-30 min; (3) wax soaking: soaking the skin tissue being subjected to the transparent treatment into a mixed liquid of dimenthylbenzene and paraffin for 20-30 min, and then soaking the skin tissue into paraffin respectively by two times, each time lasts for 45-60 min; and (4) embedding the wax-soaked skin tissue, and performing slice cutting, slice expansion, slice pasting, slice baking, dyeing and seal-fixation to obtain the skin microscopic tissue slice. The preparation method is simple in process and convenient to operate. The prepared slice is complete and shows a clear image structure.

Description

Technical field [0001] The invention relates to a preparation method of amphibian skin microscopic tissue slices, in particular to a preparation method of small salamander skin tissue slices for microscopic observation, and belongs to the technical field of microscopic tissue slices. Background technique [0002] The skin is the first natural barrier of the amphibian organism. In the process of adaptation and evolution, it plays the role of breathing, regulating water balance, ion transport, resisting predators, resisting bacteria, and fungi. These functions reflect The complexity of skin morphology characteristics and the potential physiological characteristics of skin structure play an important role in maintaining the survival of amphibians, developing and adapting to vast habitats and diverse ecological environments. [0003] Salamanders are endemic amphibians in Asia. They are also the third largest family among the 10 families of Urgodae. 66 species are known worldwide, belon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/06G01N1/28
Inventor 熊建利刘秀英孙原野张亚男胡益源吕云云孙平
Owner HENAN UNIV OF SCI & TECH
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