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Preparation method of rana kukunoris skin paraffin section

A technology for paraffin sectioning and skin, which is applied in the field of preparation of plateau wood frog skin paraffin sections, can solve the problems of poor section quality, incomplete structure, poor dyeing effect, etc., and achieve good section quality, maintain paraffin brittleness, and simple and convenient operation Effect

Pending Publication Date: 2020-06-05
QINGHAI UNIV FOR NATITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the above-mentioned deficiencies in the prior art, the present invention provides a method for preparing a paraffin section of the plateau forest frog skin. The structure of skin tissue can be clearly observed under the microscope, which effectively solves the problems of poor section quality, poor staining effect and incomplete structure

Method used

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  • Preparation method of rana kukunoris skin paraffin section

Examples

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Effect test

Embodiment 1

[0024] A preparation method for the paraffin section of the plateau Rana skin comprises the following steps in sequence:

[0025] (1) Obtain the skin of the plateau Rana forelimb, fix it in Bouin's fixative solution for 20 hours, and then cut it into a rectangular sample of 1.5mm×2.5mm for later use, so as to obtain the fixed skin;

[0026] (2) The fixed skin obtained in step (1) is transparent with 100vt% xylene for 1 time, each time for 20min, then immersed in liquid paraffin at 50°C, for 1 time, each time for 5h, then embedded in paraffin wax, and finally in Refrigerate at 3°C ​​for 10 hours;

[0027] (3) Use a Leica paraffin microtome to slice the refrigerated skin in step (2), the slice direction is perpendicular to the long axis of the body, and the slice thickness is 4-6 μm, and then bake the slices at 40°C for 22 hours;

[0028] (4) Wash the slices obtained in step (3) with 100vt% xylene for 4min, then wash the slices with absolute ethanol, 95vt% ethanol, 70vt% ethano...

Embodiment 2

[0031] A preparation method for the paraffin section of the plateau Rana skin comprises the following steps in sequence:

[0032] (1) Obtain the back skin of the plateau Rana, then fix it in Bouin's fixative solution for 24 hours, and then cut it into a rectangular sample of 1.5mm×2.5mm for later use, so as to obtain the fixed skin;

[0033] (2) The fixed skin obtained in step (1) is transparent 2 times with 100vt% xylene, each 30min, then immersed in liquid paraffin at 60°C, soaked in wax 2 times, each 6h, then embedded in paraffin, and finally in Refrigerate at 4°C for 12 hours;

[0034] (3) Use a Leica paraffin microtome to slice the refrigerated skin in step (2), the slice direction is perpendicular to the long axis of the body, and the slice thickness is 4-6 μm, and then bake the slices at 42°C for 24 hours;

[0035] (4) Wash the slices obtained in step (3) with 100vt% xylene for 5min, then wash the slices with absolute ethanol, 95vt% ethanol, 70vt% ethanol, 50vt% ethano...

Embodiment 3

[0038] A preparation method for the paraffin section of the plateau Rana skin comprises the following steps in sequence:

[0039] (1) Obtain the head skin of the plateau wood frog, fix it in Bouin's fixative solution for 26 hours, and then cut it into a rectangular sample of 1.5mm×2.5mm for later use, so as to obtain the fixed skin;

[0040] (2) The fixed skin obtained in step (1) is transparent with 100vt% xylene for 1 to 3 times, each time for 40 minutes, then immersed in liquid paraffin at 70° C., soaked in wax for 3 times, each time for 7 hours, and then embedded in paraffin wax. Finally, refrigerate at a temperature of 5°C for 14 hours;

[0041] (3) Use a Leica paraffin microtome to slice the refrigerated skin in step (2). The slice direction is perpendicular to the long axis of the body, and the slice thickness is 6 μm.

[0042] (4) Wash the slices obtained in step (3) with 100vt% xylene for 6 minutes, then wash the slices with absolute ethanol, 95vt% ethanol, 70vt% eth...

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Abstract

The invention provides a preparation method of rana kukunoris skin paraffin sections, which sequentially comprises the following steps: acquiring rana kukunoris skin, fixing in a fixing solution, andshearing to obtain fixed skin; making the fixed skin transparent by xylene, then immersing the skin in liquid paraffin for paraffin dipping and then embedding in paraffin, and finally slicing the refrigerated skin, pasting the sliced skin, and baking the pasted skin; washing the baked slices with xylene, then washing the slices with gradient ethanol, and finally washing the slices with double distilled water; dyeing the sections with hematoxylin, washing the sections with clean water, dyeing the sections with eosin, and finally washing the sections with clean water to obtain the rana kukunorisskin paraffin sections. The flaking process is simple and convenient to operate, the flaking skin paraffin section can be rapidly obtained, the section quality is good, the skin tissue structure formcan be clearly observed under an optical microscope, and the problems that the section quality is poor, the dyeing effect is poor and the structure is incomplete are effectively solved.

Description

technical field [0001] The invention relates to the technical field of section preparation, in particular to a method for preparing plateau wood frog skin paraffin sections. Background technique [0002] The plateau wood frog (Rana kukunoris) is a unique amphibian on the Qinghai-Tibet Plateau, and has adapted to the high-altitude hypoxia and strong ultraviolet natural environment of the Qinghai-Tibet Plateau. As the main organ of amphibians to resist the invasion of foreign harmful substances, regulate osmotic pressure, and maintain the stability of the internal environment, the skin is of great significance for amphibians to adapt to the ecological environment. The research on the skin structure of amphibians is mainly based on paraffin section and H.E staining technology. At present, many scholars at home and abroad have done a lot of research on the skin structure of amphibians by using paraffin section technology. Not seen in the report. Moreover, the plateau Rana skin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30G01N1/34G01N1/36
CPCG01N1/28G01N1/286G01N1/30G01N1/34G01N1/36G01N2001/305G01N2001/302
Inventor 张湑泽沈迎芳彭飞侯业茂孙玉奇
Owner QINGHAI UNIV FOR NATITIES
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