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Paraffin Section Method for Leaf and Flower Buds of Larger Explants of Carnation

A technology of paraffin sections and explants, which is applied in the preparation, sampling, and instrumentation of test samples, and can solve problems such as difficulties in the production process, difficulties in organizing xylenes, and material fragmentation

Active Publication Date: 2020-07-07
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 Section Method for Leaf and Flower Buds of Larger Explants of Carnation
  • Paraffin Section Method for Leaf and Flower Buds of Larger Explants of Carnation
  • Paraffin Section Method for Leaf and Flower Buds of Larger Explants of Carnation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Acquisition and fixation:

[0039] Take carnation leaf buds of different developmental stages, the length of the buds is 0.5~3cm, and the buds are cut into two halves from the center, and then quickly placed in FAA fixative (formalin: glacial acetic acid: 70% ethanol = 1: 1:18), fix for 24h and store in a refrigerator at 4℃;

[0040] (2) Washing and dyeing:

[0041] The material obtained in step (1) was washed 3 times with distilled water for 2 minutes each time; then placed in a 2mL centrifuge tube, stained with hematoxylin staining solution for 24 hours, washed with distilled water 3 times, 2 minutes each time, and then used at a concentration of 70v / v% Wash with ethanol for 2 times, 10min each time;

[0042] (3) Dehydration:

[0043] Dehydrate with 70v / v% ethanol for 12h, discard the ethanol, put the material in a 2mL centrifuge tube and centrifuge at 2000 rpm for 1min, replace with a new 2mL centrifuge tube, and use 85v / v% ethanol for dehydration 1.5 h, discard the eth...

Embodiment 2

[0057] (1) Acquisition and fixation:

[0058] Take carnation flower buds of different developmental stages, the length of the buds is 0.5-3cm, and cut the buds into two halves from the center, and then quickly place them in FAA fixative (formalin: glacial acetic acid: 70% ethanol = 1: 1:18), fix for 24h and store in a refrigerator at 4℃;

[0059] (2) Washing and dyeing:

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

[0061] (3) Dehydration:

[0062] Dehydrate with 70v / v% ethanol for 24h, discard the ethanol, put the material in a 2mL centrifuge tube and centrifuge at 3000 rpm for 2min, replace with a new 2mL centrifuge tube, and dehydrate with 85v / v% ethanol for 2h , Discard ethanol, cent...

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Abstract

The invention discloses a method for paraffin sectioning of larger explant leaf buds and flower buds of carnation, including material collection and fixation, cleaning and dyeing, dehydration, transparency, wax dipping and embedding, sectioning, spreading and patching, dewaxing and coverslipping and other steps. The method provided by the invention can effectively deal with the problems that the carnation explant leaf buds and flower buds are too large, slices are incomplete, discontinuous, and have cavities. It can well fix the various stages of tissue development, can observe the whole process of leaf bud and flower bud development, and is easy to operate, which has good development and application value.

Description

Technical field [0001] The invention relates to a paraffin sectioning method for plants, in particular to a paraffin sectioning method for leaf buds and flower buds of larger explants of carnation. Background technique [0002] Paraffin section is the main method used to observe the morphological structure of cell tissues, and it has also been widely used in research in many disciplines. There are many procedures and links in paraffin section production, and the cycle is long. Each link will affect the quality of the section. Common problems such as broken materials, incomplete sections, discontinuities, and voids. [0003] The cause of material fragmentation can be traced back to many steps in the paraffin production process, including dehydration, transparency, wax penetration, embedding, sectioning, dewaxing, and sealing, etc. As long as one of the steps is wrong, the material may be broken. Among them, the steps of dehydration, transparency, wax immersion, embedding, and dewax...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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