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Pine needle sectioning observation method

A technology of needles and pine trees, which is applied in the field of filming and observation of pine needles. It can solve the problems of high scraping operation requirements, difficulty in observing clearly, damage to leaf skin, etc., and achieves shortening filming time, clear display, and easy purchase. Effect

Inactive Publication Date: 2010-07-07
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commonly used direct scraping method is to place the blade on the surface of a glass slide or other hard objects, and scrape off the epidermis and mesophyll cells on the upper side of the leaf epidermis with a single-sided or double-sided blade. Unfavorable factors such as softness and unevenness, the scraping operation of the leaf epidermis requires high requirements. Even if it is carefully operated, it often damages the leaf epidermis and destroys its integrity
When ordinary optical microscopes and electron microscopes observe the leaf epidermis and structural features of gymnosperms, some features are difficult to observe clearly, and there are also some difficulties and problems in the production of ordinary optical microscope observations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The method for making sheets of the pine needle leaf epidermis of the present embodiment comprises the following steps

[0037] (1) Take off a piece of needle leaf, dip it in water and paste it on the surface of laundry soap, hold the laundry soap in your left hand or place it on the table, press the back of the needle leaf with your left thumb, and hold a single-sided blade in your right hand, after dipping in water , tilt the single-sided blade, and gently scrape off the upper part of the leaf along the leaf surface at an appropriate distance from the leaf tip (or the front end of the leaf segment). After scraping 1-3 times, re-wet the blade with water, and Remove the residue attached to the blade, and then continue to gently scrape off the upper part of the leaf with the blade, scrape off the upper epidermis and mesophyll cells of the leaf as much as possible, leaving only the lower epidermis of the leaf, laundry soap When the upper blade is gently scraped by the sin...

Embodiment 2

[0042] The method for making sheets of the pine needle leaf epidermis of the present embodiment (in order to observe the stomata profile) comprises the following steps

[0043] Step (1) and step (2) are the same as step (1) and step (2) in Example 1 for slicing and cleaning steps of the coniferous leaf epidermis;

[0044] (3) Transfer the washed leaf epidermis to a glass slide, and add dropwise a mixture of absolute ethanol and cedar oil (the volume ratio of absolute ethanol to cedar oil is 50:1), so that it completely covers the surface of the coniferous slices , use absorbent paper to absorb the white liquid that diffuses to the surroundings; when the mixed solution is almost volatilized, add the mixed solution of absolute ethanol and cedar oil dropwise, repeat 5 times; then add the mixed solution of absolute ethanol and cedar oil dropwise, and then Cover with a coverslip;

[0045] (4) Place the finished glass slide on the microscope stage, observe and take pictures under t...

Embodiment 3

[0047] The sheet-making method of the pine needle leaf cross-section of the present embodiment comprises the following steps

[0048] (1) Pick off a piece of needle, put it on the surface of the laundry soap, put the laundry soap on the table, hold the single-sided blade in your right hand, and use the edge of the blade to cut the needle from the left side at the place where the needle needs to be cut To the right, cut a cross-cut slice;

[0049] (2) Use pointed tweezers to transfer it from the laundry soap to a petri dish filled with water, stir and clean it with a brush, and change the water for 3 times;

[0050] (3) Put the washed leaf cross section into 1% safranin aqueous solution and stain for 10 minutes; For a mixed solution of absolute ethanol and cedar oil (the volume ratio of absolute ethanol to cedar oil is 60:1), use absorbent paper to wipe off the white liquid that diffuses around; Cedar oil mixture, after repeating 6 times, add cedar oil dropwise on the leaf ep...

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Abstract

The invention relates to a pine needle sectioning observation method which includes slicing, washing, transparent sectioning and microscopic observation, also can include the dying step. Compared with the direct scraping method in prior art, the invention adopts the method of directly placing plant material on the surface of a laundry soap for slicing, has simpler operation, quicker sectioning speed and higher efficiency, and is particularly applicable to the cuticle sectioning of harder leaves; as xylene with toxic and side effects is replaced by cedar oil as clearing reagent, the researchers are protected, the reagent is easy to purchase, the sectioning time is also reduced and the experiment cost is reduced; a common lighting source is combined with an ultraviolet source for fluorescence microscopy and photography, and the cell contour which cannot be observed only through fluorescence source can be observed by the combined light source, and the structural information unavailable under fluorescence is acquired as supplement.

Description

technical field [0001] The invention relates to a method for making slices and observing pine needles, and belongs to the field of plant microscopy technology. Background technique [0002] The leaf epidermis and structural characteristics of plants are determined by the genetic nature of plants and are affected by environmental factors, which can reflect the systematic relationship and growth status among plant groups to a certain extent. Fossil identification, plant evidence and breeding, and even bionics and aesthetics have extremely important research values. [0003] There are many methods for making plant leaf epidermis. According to the main methods used, they can be roughly divided into direct tearing method, direct scraping method, imprinting method, scotch tape tearing method, chromic acid isolation method, sodium hypochlorite method, etc. The scraping method can be roughly divided into: (1) direct scraping method, that is, adopting physical methods, the leaf epid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/34G01N1/30G01N21/64G01N21/84
Inventor 洪亚平
Owner HENAN UNIV OF SCI & TECH
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