Method for greatly improving quality of scanning electron microscope sample of salivary sheath of rice planthopper

A scanning electron microscope, sample quality technology, applied in the field of agricultural science, can solve problems such as invisible samples, unclear details, dry cracks, etc.

Active Publication Date: 2019-04-12
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

All three kinds of rice planthoppers directly suck rice damage, or spread and induce rice diseases, causing great losses to my country's rice production every year
However, the scanning electron microscope has high requirements for the processing of saliva sheath samples. After the previous research, the researchers proposed a method for preparing the saliva sheath electron microscope samples of the rice planthopper. However, in the actual operation of the researchers, there are still many Imperfect places lead to the observation of rice planthopper saliva sheath scanning sample pictures showing problems such as no samples, dry cracks, defects, breaks, and unclear details, which seriously affect the progress of our scientific research on rice planthopper control

Method used

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  • Method for greatly improving quality of scanning electron microscope sample of salivary sheath of rice planthopper
  • Method for greatly improving quality of scanning electron microscope sample of salivary sheath of rice planthopper
  • Method for greatly improving quality of scanning electron microscope sample of salivary sheath of rice planthopper

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Experimental program
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Effect test

Embodiment 1

[0030] 1. Sampling: Take about 20 4-5 instar rice planthopper nymphs and feed them for 24 hours at 25°C, relative humidity of 60%, and photoperiod of 16:8 (D:L).

[0031] Taking SBPH as an example, under the above temperature and humidity conditions: Take a glass tube with a diameter of 2.5 cm at both ends, seal one end with gauze, and seal the other end with double-layer parafilm (the two layers of film contain artificial feed for rice planthopper) , put the rice planthopper into the glass tube. After feeding for 24 hours, uncover the double-layer parafilm membrane to obtain the parafilm membrane with saliva sheath;

[0032] Table 1 The relationship between the number of rice planthopper nymphs, feeding time and whether salivary sheaths can be observed in the artificial diet:

[0033]

[0034] 2. Cleaning of the sample: wash the parafilm membrane with the saliva sheath with PBS buffer solution (NaCl 137mmol / L, KCl2.7mmol / L, NaCl 2 HPO 4 10mmol / L, KH 2 PO 4 2mmol / L, ...

Embodiment 2

[0049] 1. Sampling: Take about 30 4-5 instar rice planthopper nymphs and feed them for 24 hours at 25°C, relative humidity of 60%, and photoperiod of 16:8 (D:L). Take a glass tube with a diameter of 2.5 cm at both ends, seal one end with gauze, and seal the other end with double-layer parafilm (the two layers of film contain artificial feed for rice planthoppers), and put rice planthoppers into the glass tube. After feeding for 24 hours, uncover the double-layer parafilm membrane to obtain the parafilm membrane with saliva sheath.

[0050] 2. Cleaning of the sample: wash the parafilm membrane with the saliva sheath in 75% ethanol solution diluted with phosphate buffer solution (PBS) for 1 hour, and replace the cleaning solution every 15 minutes. After that, place in PBS and wash twice, 5min each time.

[0051] 3. Fixation of the sample: the fixation ratio of the rice planthopper saliva sheath fixation solution is 4% paraformaldehyde: 2.5% glutaraldehyde = 3:1, fix at 4°C for ...

Embodiment 3

[0060] 1. Sampling: Take about 25 4-5 instar rice planthopper nymphs and feed them for 24 hours at 25°C, relative humidity 60%, and photoperiod 16:8 (D:L).

[0061] 2. Cleaning of the sample: wash the parafilm membrane with the saliva sheath in 75% ethanol solution diluted with phosphate buffered solution (PBS) for 1 hour, and replace the cleaning solution every 15 minutes. After that, place in PBS and wash twice, 5min each time.

[0062] 3. Fixation of the sample: the fixative ratio of the rice planthopper saliva sheath is 4% paraformaldehyde: 2.5% glutaraldehyde = 2.5:1, fixed at 4°C for 1 hour; or choose a fixative solution of 4% paraformaldehyde: Qular Through X-100=12:1, fixed overnight at 4°C. After the fixation was selected, the fixative was sucked out, rinsed with phosphate buffered saline (PBS) for 1 h, and the medium was changed every 20 min.

[0063] 4. Dehydration of samples: suck out phosphate buffer solution (PBS), add 15% ethanol solution to balance for 1 hour...

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Abstract

The invention discloses a method for greatly improving the quality of a scanning electron microscope sample of the salivary sheath of rice planthopper. The method comprises the following steps of: 1)taking a sample; 2) washing; 3) fixing; 4) dehydrating; 5) replacing ethanol; 6) drying; 7) pasting the sample and coating; 8) observing the sample. According to the method, a complete salivary sheathimage of the rice planthopper with a high resolution and a smooth surface is obtained by scanning electron microscope observation. A plurality of key steps in the preparation of the scanning electronmicroscope sample of the salivary sheath of the rice planthopper are improved, so that the problems of invisible samples, dry cracks, defects, fractures, unclear details and the like that are easy toappear in the scanning electron microscope observation of the salivary sheath of the rice planthopper are solved. The workload of relevant experimental personnel is reduced and the efficiency is improved.

Description

technical field [0001] The invention relates to the field of agricultural science and technology, in particular to a method for greatly improving the quality of rice planthopper saliva sheath scanning electron microscope samples. Background technique [0002] Rice planthoppers are an important class of agricultural pests belonging to the order Homoptera, Planthoppers family, including the brown planthopper Nilaparvatalugens The white-backed planthopper Sogatella furcifera (Horváth) and the white-backed planthopper Laodelphax striatellus (Fallén). All three kinds of rice planthoppers directly suck rice damage, or spread and induce rice diseases, causing great losses to my country's rice yield every year. It is precisely because of the great harm of rice planthoppers that the state has allocated a large amount of funds for the prevention and control of three kinds of rice planthoppers for many years. Feeding habits are a very important link in the study of rice planthoppers'...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/2202
CPCG01N23/2202
Inventor 鞠佳菲孙洋刘宝生方继朝纪锐
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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