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Paraffin slicing method for large explant leaf-buds and flower-buds of carnation

A paraffin section and explant technology, which is applied in the preparation, sampling, measuring device and other directions of samples for testing, can solve the problems of difficult xylene tissue, increased brittleness of materials, fragile tissue and other problems, and achieves good development and application value. Extend the wax soaking time and complete the sectioning effect

Active Publication Date: 2018-02-02
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The carnation leaf buds and flower buds are made of very large materials, and the length of the buds can reach about 2 to 3 centimeters, so the production process will bring considerable difficulties
First of all, the dehydration process is not easy. Because the material is too large and the dehydration is not complete, a hollow wax sheet will be formed. Secondly, there will be problems in the transparent process. It is difficult for xylene to enter the tissue, but it cannot be achieved by simply extending the xylene soaking time. Purpose, because the soaking time is too long, it will have adverse effects on the material, which will increase the brittleness of the material, and the tissue will be brittle, etc.
Therefore, the paraffin sectioning of larger buds often causes the material to break, resulting in the failure of sectioning.

Method used

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  • Paraffin slicing method for large explant leaf-buds and flower-buds of carnation
  • Paraffin slicing method for large explant leaf-buds and flower-buds of carnation
  • Paraffin slicing method for large explant leaf-buds and flower-buds of carnation

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Experimental program
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Effect test

Embodiment 1

[0038] (1) Material collection and fixation:

[0039] Take carnation leaf buds of different development stages, the length of the buds is 0.5-3cm, and cut the buds in half from the center lengthwise, and then quickly place them in FAA fixative solution (formalin: glacial acetic acid: 70% ethanol = 1: 1:18), fixed for 24h, and stored in a refrigerator at 4°C;

[0040] (2) Cleaning and dyeing:

[0041] Wash the material obtained in step (1) with distilled water for 3 times, each time for 2 minutes; then place it in a 2mL centrifuge tube, stain with hematoxylin dye solution for 24 hours, wash with distilled water for 3 times, each time for 2 minutes, and then use a concentration of 70v / v% Washed twice with ethanol, 10min each time;

[0042] (3) Dehydration:

[0043] Dehydrate with ethanol with a concentration of 70v / v% for 12h, discard the ethanol, put the material into a 2mL centrifuge tube and centrifuge at 2000 rpm for 1min, replace with a new 2mL centrifuge tube, and dehyd...

Embodiment 2

[0057] (1) Material collection and fixation:

[0058] Take carnation flower buds of different development stages, the length of the buds is 0.5-3cm, and cut the buds in half from the center, and then quickly place them in FAA fixative solution (formalin: glacial acetic acid: 70% ethanol = 1: 1:18), fixed for 24h, and stored in a refrigerator at 4°C;

[0059] (2) Cleaning and dyeing:

[0060] Wash the material obtained in step (1) with distilled water for 3 times, each time for 5 minutes; then place it in a 2mL centrifuge tube, stain with hematoxylin dye solution for 24 hours, wash with distilled water for 3 times, each time for 5 minutes, and then use a concentration of 70v / v% Washed twice with ethanol, 10min each time;

[0061] (3) Dehydration:

[0062] Dehydrate with ethanol with a concentration of 70v / v% for 24h, discard the ethanol, put the material into a 2mL centrifuge tube and centrifuge at 3000 rpm for 2min, replace with a new 2mL centrifuge tube, and dehydrate with...

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Abstract

The invention discloses a paraffin slicing method for large explant leaf-buds and flower-buds of carnation, comprising steps as follows: material selection and fixation, cleaning and dyeing, dehydration, transparency, paraffin impregnation and embedding, slicing, section flatting and section sticking, deparaffinating and section sealing, etc. By the method of the invention, problems such as incomplete and discontinuous sections, cavity and the like due to overlarge explant leaf-buds and flower-buds of carnation can be effectively solved. Various growth stages of tissues can be greatly fixed, and the whole growth process of leaf buds and flower buds can be observed. In addition, the method is simple to operate and has good development and application value.

Description

technical field [0001] The invention relates to a method for plant paraffin sectioning, in particular to a method for paraffin sectioning large explant leaf buds and flower buds of carnation. Background technique [0002] Paraffin section is the main method for observing the morphological structure of cell tissue, and it has been widely used in research in many disciplines. There are many procedures and links in the production of paraffin slices, and the cycle is long. Each link will affect the quality of slices. Common problems include broken materials, incomplete slices, discontinuities, and voids. [0003] The cause of material breakage can be traced back to many steps in the paraffin wax production process, including dehydration, transparency, paraffin penetration, embedding, sectioning, dewaxing and sealing, etc., as long as one of the steps goes wrong, the material may be broken. Among them, the steps of dehydration, transparency, wax immersion, embedding, and dewaxin...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30G01N1/36
CPCG01N1/286G01N1/30G01N1/36G01N2001/2873
Inventor 周旭红瞿素萍海玉芬蒋亚莲桂敏莫锡君
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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