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Method for making paraffin slices of Xanthos sorbifolium cutting branches and their calluses

A technology for cutting branches and callus, which is applied in the field of paraffin slices of cutting branches and callus of Xanthos sorbifolium, can solve the problems of callus deformation and tearing, difficult organizational structure, incomplete dehydration, etc., and achieve Clear organizational structure, shortened softening time, good result stability

Active Publication Date: 2021-06-25
HENAN UNIV OF ANIMAL HUSBANDRY & ECONOMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] X. sorbifolium is a highly lignified material, but the callus produced is a relatively young part, and it is extremely difficult to obtain a complete tissue section of both
It is more difficult for highly lignified tissue reagents to enter, but the goal cannot be achieved by simply prolonging the soaking time of the reagents, because the soaking time is too long, which will easily cause deformation and tearing of the callus
If the soaking time is too short, the dehydration will not be complete, and the problem of hollow slices often occurs during the production process. If the soaking time is too long, it will easily cause the material to harden. It is quite difficult to obtain a complete tissue structure.

Method used

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  • Method for making paraffin slices of Xanthos sorbifolium cutting branches and their calluses
  • Method for making paraffin slices of Xanthos sorbifolium cutting branches and their calluses
  • Method for making paraffin slices of Xanthos sorbifolium cutting branches and their calluses

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Embodiment 1

[0064] The invention provides a method for making paraffin slices of Xanthos sorbifolium cuttage branches and calluses thereof, comprising:

[0065] 1) Obtaining materials and fixing: cut the stem section containing callus from the cutting branch of X. sorbifolium, the length of the stem section is 0.4cm; soak the stem section in the fixative solution, vacuumize to 0.5 atmospheric pressure, and keep it for 5 minutes , and then stand at 4°C for 24 hours;

[0066] The fixative is: formaldehyde with a mass fraction of 38%, glacial acetic acid with a mass fraction of 100%, glycerol with a mass fraction of 100%, and ethanol with a mass fraction of 50% in a volume ratio of 1:1:1: 12 Prepared after thorough mixing.

[0067] 2) Rinse: the stem section after the material is fixed is repeatedly washed three times with 50% ethanol, and the washing time is 30 seconds once, wherein, the stem section is taken from the cutting branch of X. sorbifolium containing callus;

[0068] 3) softeni...

Embodiment 2

[0093] The invention provides a method for making paraffin slices of Xanthos sorbifolium cuttage branches and calluses thereof, comprising:

[0094] 1) Obtaining materials and fixing: cut the stem section containing callus from the cutting branch of X. sorbifolium, the length of the stem section is 0.6cm; soak the stem section in the fixative solution, vacuumize to 0.8 atmosphere, and keep it for 5 minutes , and then stand at 4°C for 24 hours;

[0095] The fixative is: formaldehyde with a mass fraction of 38%, glacial acetic acid with a mass fraction of 100%, glycerol with a mass fraction of 100%, and ethanol with a mass fraction of 50% in a volume ratio of 1:1:1: 14 Prepared after thorough mixing.

[0096] 2) Rinse: the stem section after the material is fixed is repeatedly washed three times with 50% ethanol, and the washing time is 30-60 seconds once, wherein, the stem section is taken from the cutting branch of Xanthos sorbifolium containing callus;

[0097] 3) softening...

Embodiment 3

[0121] The invention provides a method for making paraffin slices of Xanthos sorbifolium cuttage branches and calluses thereof, comprising:

[0122] 1) Obtaining materials and fixing: cut the stem segment containing callus from the cutting branch of X. sorbifolium, the length of the stem segment is 0.5cm; soak the stem segment in the fixative solution, vacuumize to 0.6 atmospheric pressure, and keep it for 5 minutes , and then stand at 4°C for 24 hours;

[0123] The fixative is: formaldehyde with a mass fraction of 38%, glacial acetic acid with a mass fraction of 100%, glycerol with a mass fraction of 100%, and ethanol with a mass fraction of 50% in a volume ratio of 1:1:1: 17 Prepared after thorough mixing.

[0124] 2) Rinse: the stem section after the material is fixed is repeatedly washed three times with 50% ethanol, and the washing time is 30-60 seconds once, wherein, the stem section is taken from the cutting branch of Xanthos sorbifolium containing callus;

[0125] 3)...

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Abstract

The invention discloses a method for making paraffin slices of Xanthos sorbifolium cuttage branches and calluses thereof, comprising: taking materials and fixing them, washing, softening, dehydrating, transparentizing, dipping in wax and embedding, slicing, sticking and unfolding, removing Waxing and dyeing, and sealing; the specific method of washing is: repeatedly wash the stem section after getting the materials and fixing it three times with 50% ethanol; Hydrofluoric acid, glycerol with a mass fraction of 50% at 50° C. and a softening solution are soaked for a certain period of time; wherein, the softening solution is an equal volume mixture of alcohol with a mass fraction of 50% and glycerol with a mass fraction of 100%. The method of the present invention has the advantages of easy operation, good stability of results, convenience for a large number of basic researches, complete slices, uniform thickness, clear tissue structure, good dyeing contrast, complete cell shape, and the obtained slices of X. sorbifolium callus have good Continuity, can be preserved forever.

Description

technical field [0001] The invention relates to the technical field of paraffin sectioning, in particular to a paraffin sectioning method for cutting branches of Xanthos sorbifolium and callus thereof. Background technique [0002] Paraffin slices are an important and most commonly used method for observing plant tissue structure, especially one of the important technical means for observing plant callus. The main advantages of paraffin slices are easy operation, uniform and extremely thin slices, and wax bands. The continuity is easy to make serial slices and can be preserved permanently. Xanthoceras sorborifolia bunge belongs to the genus Xanthoceras sorborifolia, a deciduous tree or a large shrub, and its cutting propagation is a tree species that is difficult to root. The callus and cuttings during the rooting process of Xanthoceras sorborifolia were observed by paraffin slices The structural changes of basal stems can theoretically clarify the location of adventitious ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N1/30G01N1/36
CPCG01N1/28G01N1/286G01N1/30G01N1/36G01N2001/2873
Inventor 宗建伟杨雨华杨军王鹏张馥郁左金淼胡华锋程倩李艳刘策王帅博
Owner HENAN UNIV OF ANIMAL HUSBANDRY & ECONOMY
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