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Formula and method for immobilizing tick sample for scanning electron microscopy

A fixing method and scanning electron microscope technology, applied in the preparation of test samples, etc., can solve problems such as poor penetration ability, and achieve the effect of less time consumption and uniform coating

Inactive Publication Date: 2016-04-13
XINJIANG AGRI UNIV
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention: to provide a tick sample scanning electron microscope formula and method to solve the problem of poor penetrating ability of fixatives and methods of conventional formulas

Method used

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  • Formula and method for immobilizing tick sample for scanning electron microscopy
  • Formula and method for immobilizing tick sample for scanning electron microscopy
  • Formula and method for immobilizing tick sample for scanning electron microscopy

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Embodiment Construction

[0016] Example 1. Twelve male adult ticks were checked to see if there was any residual host tissue in the false head base, and if they were removed, they were removed with fine surgical forceps. Carefully wash the body surface of ticks with PBS buffer (pH=7.2), and then use ultrasonic cleaning for 60 minutes. Divide the cleaned ticks into four groups, three in each group, immerse the first and second groups of ticks in PBS buffer, lightly puncture 3 small holes under the 1st to 3rd basal segments of the ticks under the dissecting microscope, At room temperature, fix the treated tick samples with 1.5% glutaraldehyde fixative for 3 hours, 2.5% glutaraldehyde fixative for 4 hours, and 3% glutaraldehyde fixative for 5 hours. Soak the third and fourth groups of ticks in distilled water, lightly puncture 3 small holes below the 1-3 basal segments of the ticks under a dissecting microscope, and put them in 2.5% glutaraldehyde fixative solution (fixed with no new formula) Solution),...

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Abstract

The invention relates to a formula and method for immobilizing a tick sample for scanning electron microscopy, which belongs to the technical field of methods for immobilizing samples for scanning electron microscopy. The immobilization formula comprises a PBS buffer solution, polysorbate and glutaraldehyde, and a glutaraldehyde immobilization liquid is prepared from the above-mentioned components. The method comprises the following steps: (1) observing whether host tissue is left in basis capituli of the tick sample and if so, removing the host tissue; (2) cleaning the surface of the tick sample; (3) immobilizing the tick sample at room temperature by using the glutaraldehyde immobilization liquid; (4) carrying out gradient dehydration with ethanol on the immobilized tick sample; (5) putting the tick sample into mixed liquor of absolute ethyl alcohol and acetone for displacement, and then putting the tick sample into acetone for displacement; and (6) drying the tick sample in a vacuum freeze drying instrument for drying and spraying gold on the tick sample. During scanning electron microscopic observation, the phenomenon of fuzzy and foggy images or incapable imaging due to ionization discharging of the sample as water vapor produced after electron beam bombardment of the sample encounters high-energy electron streams is prevented.

Description

technical field [0001] The invention belongs to the technical field of methods for fixing parasite scanning electron microscope samples, and in particular relates to a formula and method for fixing tick samples for scanning electron microscope. Background technique [0002] Ticks are also called ticks, which have many local common names, such as grass scorpion, dog bean, grass crawler, grass tick, paederus, etc. A vertebrate arthropod that feeds on its blood for survival. The transmission and harm of pathogens are second only to mosquitoes, and they are also storage hosts and transmission media for various diseases in animals and humans. Ticks release toxins when they bite humans and animals to suck blood, which can cause paralysis, irritation, toxic reactions and allergic reactions of the host. In my country, ticks are vectors of various pathogens, which can cause Q fever, babesiosis, theileriasis, forest encephalitis, Xinjiang hemorrhage and Lyme disease, etc., posing a ...

Claims

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Application Information

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IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 巴音查汗·盖力克吴辉张扬翟少华郭庆勇陈千林明·乌尔娜李永畅伊春阳朱玉涛史亚明
Owner XINJIANG AGRI UNIV
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