Paraffin slicing method of tree stem tissue

A technology for paraffin sectioning and tissue, which is applied in the field of plant microtissue sectioning, can solve the problems of tissue fragility, achieve high differentiation, improve production efficiency, and simplify steps

Active Publication Date: 2017-03-22
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The object of the present invention is to provide a method for paraffin sectioning of tree stem tissue, to solve the technical problem that the phloem, cambium and xylem in the stem tissue are fragile during the slicing process due to inconsistent hardness

Method used

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  • Paraffin slicing method of tree stem tissue
  • Paraffin slicing method of tree stem tissue
  • Paraffin slicing method of tree stem tissue

Examples

Experimental program
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Effect test

Embodiment 1

[0075] The paraffin section method of tree stem tissue in the present embodiment, concrete steps are as follows:

[0076] (1) Stem tissue sampling:

[0077] In the field, use micro-growth cones (Trephor, cambium sampler) to collect stem tissue samples at the DBH (about 1.3m) of white skewers (Picea meyeri Rebd.EtWils). The tissue size is 2mm in diameter and 18mm in length, including phloem, cambium and 4-year xylem regions;

[0078] (2) Tissue fixation:

[0079] After sampling, stem tissue is put into the 7mL penicillin bottle that fills universal fixative (volume ratio 70% ethanol:formaldehyde:acetic acid=9:0.5:0.5) rapidly (the volume ratio of the amount of fixative to stem tissue is greater than 20:1), use a syringe to evacuate the stem tissue completely to the bottom of the bottle, and then put it in a 5°C incubator for refrigerated storage for 1 week. During the storage process, keep the sample bottle in an upright state;

[0080] (3) Primary softening:

[0081] Put t...

Embodiment 2

[0128] In the paraffin section method of the stem tissue of the tree in the present embodiment, the stem tissue at the breast diameter of the North China larch (Larixprincipis-rupprechtii Mayr.) is collected in step (1), and the embedding block after trimming is placed in the step (7). Enter 30 ℃ of constant temperature water baths and soften 2d, other operations are all the same as embodiment 1.

[0129] The micrograph of the cross-section of the stem tissue of North China larch (wood hardness is grade III) prepared above is shown in figure 1 . In the figure, 1 is the phloem, 2 is the cambium, and 3 is the xylem, and the scale bar in the figure is 100 μm.

Embodiment 3

[0131] The paraffin section method of tree stem tissue in the present embodiment, except collecting cork oak (Quercus variabilis Bl.) stem tissue at breast diameter place in step (1) (because the cork oak bark is thicker, need suitably strip off the outer bark), step In (7), the embedding block after trimming was placed in a 30°C constant temperature water bath to soften for 7 days, and other operations were the same as in Example 1.

[0132] The micrograph of the cross-section of the stem tissue of the cork oak (wood hardness is Ⅳ~Ⅴ grade) prepared above is shown in figure 1 . In the figure, 1 is the phloem, 2 is the cambium, and 3 is the xylem, and the scale bar in the figure is 100 μm.

[0133] Explanation: The "%" related to ethanol in the text refers to the volume fraction, and the "%" in the 95% ethanol solution of 1% safranin and 0.5% fast green refers to the mass fraction.

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Abstract

The invention discloses a paraffin slicing method of tree stem tissue, and belongs to the technical field of plant microscopic structure slicing. The paraffin slicing method of the tree stem tissue comprises the operations of tissue fixing, primary softening, dehydration, transparency, waxing, embedding, secondary softening, slicing, slice folding, slice gluing, slicing roasting, dewaxing, rehydration, dyeing, slice sealing and the like, wherein the addition of softening treatment can solve the problem that phloem, a forming layer and xylem in the stem tissue are easy to break in the slicing process due to inconsistent hardnesses. The softening treatment comprises the steps of firstly after the tissue is fixed, adding the operation of using ethyl alcohol/glycerinum mixed liquor to soften the tissue; secondly after embedding and coarse machining, adding the operation of using warm water to soak so as to soften the tissue to be sliced. Experiments demonstrate that a softened tissue can lower probability that the stem tissue is easy to break during slicing, and improve slicing success rate.

Description

technical field [0001] The invention relates to a method for paraffin sectioning tree stem tissues, and belongs to the technical field of plant microscopic tissue sectioning. Background technique [0002] Paraffin section is the most widely used in conventional histological production techniques, mainly used for the observation of the morphology of cell tissue, and the prepared sections are made into permanent sections after sealing, which can be stored for a long time. [0003] The paraffin section technique of tree stem tissue is an important experimental technique for studying the radial growth and physiological and ecological mechanism of trees. Tree stem tissue includes three parts: xylem, cambium and phloem, and the hardness of these three parts is inconsistent. The xylem is usually hard and has a thick cell wall; the cells of the cambium and elongated cells are soft, have a high water content, and have thin cell walls; and the cell walls of the phloem are relatively ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/28G01N1/30
Inventor 徐军亮章异平苏维许晓利朱文琰
Owner HENAN UNIV OF SCI & TECH
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