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Rapid sample preparation method of young plants for scanning electron microscopy

A scanning electron microscope, a young and tender technology, applied to the use of wave/particle radiation for material analysis, measuring devices, instruments, etc., can solve the complicated and lengthy sample preparation process, and achieve simple and easy operation, not easy to absorb water, and prevent shrinkage deformation effect

Inactive Publication Date: 2019-11-22
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the above methods, there are also drying methods such as natural drying, drying drying, critical point drying, vacuum drying, etc. These drying methods have their own advantages and disadvantages, and are suitable for different types of biological samples. After this treatment, biological samples can be obtained Clear images of surface topography, but there are also many defects at the same time. The main performance is that the sample preparation process is complicated and tedious. On the other hand, after a series of treatments, the sample itself will shrink and deform to varying degrees, especially young and aquatic plants

Method used

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  • Rapid sample preparation method of young plants for scanning electron microscopy
  • Rapid sample preparation method of young plants for scanning electron microscopy
  • Rapid sample preparation method of young plants for scanning electron microscopy

Examples

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Embodiment 1

[0026] The present embodiment is an example of the method for sample preparation of young plants for scanning electron microscope observation according to the present invention, comprising the following steps:

[0027] (1) Take Dibaolan Herui column and place it in a test tube equipped with FAA for fixation for more than 24 hours. The fixative solution is prepared with 50% ethanol.

[0028] (2) Place the fixed material step by step in 30% ethanol→40% ethanol→50% ethanol→55% ethanol→60% ethanol→65% ethanol→70% ethanol→75% ethanol→80% ethanol→85% Ethanol → 90% ethanol → 95% ethanol → 100% ethanol for dehydration. Each step is to use a vacuum pump to pump air for 20 minutes, and let it stand for 30 minutes under natural conditions in the room.

[0029] (3), put the dehydrated material gradually in 2 / 3 absolute ethanol + 1 / 3 pure To transparent agent → 1 / 2 absolute ethanol + 1 / 2 pure To transparent agent → 1 / 3 absolute alcohol + 2 / 3 pure To transparent agent → pure To transpare...

Embodiment 2

[0032] The present embodiment is another example of the method for sample preparation of young plants for scanning electron microscope observation according to the present invention, comprising the following steps:

[0033] (1) Take a certain size of banana callus and place it in a test tube equipped with FAA to fix it for more than 24 hours. The fixative is prepared with 50% ethanol.

[0034] (2) Place the fixed material step by step in 30% ethanol→40% ethanol→50% ethanol→55% ethanol→60% ethanol→65% ethanol→70% ethanol→75% ethanol→80% ethanol→85% Ethanol → 90% ethanol → 95% ethanol → 100% ethanol for dehydration. Each step is to use a vacuum pump to pump air for 20 minutes, and let it stand for 30 minutes under natural conditions in the room.

[0035] (3), put the dehydrated material gradually in 2 / 3 absolute ethanol + 1 / 3 pure To transparent agent → 1 / 2 absolute ethanol + 1 / 2 pure To transparent agent → 1 / 3 absolute alcohol + 2 / 3 pure To clearer → pure To clearer I → pure...

Embodiment 3

[0038] The present embodiment is another example of the method for sample preparation of young plants for scanning electron microscope observation according to the present invention, comprising the following steps:

[0039] (1) Put a section of seaweed into a test tube equipped with FAA and fix it for more than 24 hours, and the fixing solution is prepared with 50% ethanol.

[0040] (2) Place the fixed material step by step in 30% ethanol→40% ethanol→50% ethanol→55% ethanol→60% ethanol→65% ethanol→70% ethanol→75% ethanol→80% ethanol→85% Ethanol → 90% ethanol → 95% ethanol → 100% ethanol for dehydration. Each step is to use a vacuum pump to pump air for 20 minutes, and let it stand for 30 minutes under natural conditions in the room.

[0041] (3), put the dehydrated material gradually in 2 / 3 absolute ethanol + 1 / 3 pure To transparent agent → 1 / 2 absolute ethanol + 1 / 2 pure To transparent agent → 1 / 3 absolute alcohol + 2 / 3 pure To clearer → pure To clearer I → pure To clearer...

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Abstract

A rapid sample preparation method of young plants for scanning electron microscopy includes the following steps: taking a fresh plant material and soaking the fresh plant material in FAA fixative solution for more than 24h; dehydrating the fixed material with a graded series of alcohol; and gradually placing the dehydrated material in a reagent prepared from anhydrous ethanol and a pure TO transparent agent in different proportions for transparency. In each step of dehydration and transparency, air is pumped for 20 minutes by a vacuum air pump, and the material is allowed to stand for 30 minutes under indoor natural conditions. After the last air pumping, the material is taken out of the pure TO transparent agent and put in a culture dish for standing until the TO transparent agent in thematerial volatilizes completely, and then, the material can be put under a scanning electron microscope for observation. The method can better maintain the cell morphology of different plants, including mature and tender tissues.

Description

technical field [0001] The invention belongs to the technical field of electron microscope sample preparation methods, and relates to a rapid sample preparation method for young plants for scanning electron microscope observation. Background technique [0002] It is one of the important means of biological research to observe the tiny morphological features on the surface of biological samples by using scanning electron microscopy to obtain more detailed and subtle structural features than those observed by optical microscopy. The preparation process of the sample is the key to obtaining true, clear and ideal scanning electron microscope observation results. Samples used for conventional scanning electron ytterbium observation must be thoroughly dried. Samples with high water content will cause adverse consequences in the vacuum lens barrel of the scanning electron microscope, such as blurred images, foggy appearance, or no imaging at all; or prone to morphological damage i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/2202G01N23/2251
CPCG01N23/2202G01N23/2251G01N2223/07G01N2223/102G01N2223/612
Inventor 周琼廖天蓝裴艳艳任惠辛佳佳蒙思悦伍玉玲黄朝戟李刚
Owner GUANGXI UNIV
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