CN107421793A - The method that Predatory Mites reproductive system is observed under ESEM - Google Patents
The method that Predatory Mites reproductive system is observed under ESEM Download PDFInfo
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- CN107421793A CN107421793A CN201710808937.5A CN201710808937A CN107421793A CN 107421793 A CN107421793 A CN 107421793A CN 201710808937 A CN201710808937 A CN 201710808937A CN 107421793 A CN107421793 A CN 107421793A
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- mite
- male
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- predatory mites
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2202—Preparing specimens therefor
Abstract
The present invention relates to the method that Predatory Mites reproductive system is observed under ESEM, it is characterized in that the female mite of Phytoseiidae Predatory Mites, male mite in first being mated, soaked, be dehydrated by lactic acid again, dried, viscous platform, metal spraying are handled, the method that reproductive system is finally observed under ESEM.The present invention obtains Predatory Mites reproductive system first using very simple cleverly method, and the purpose that the trickle external structure of reproductive system is observed under ESEM is realized first, the acquisition of reproductive system can provide raw material to observe its structure under ESEM, and the result of ESEM is advantageous to the research to reproductive system.
Description
Technical field
The present invention relates to the method that Predatory Mites reproductive system is observed under ESEM, belong to Predatory Mites Basic of Biology and grind
Study carefully field.
Background technology
Research shows that the Sex Determination period of Phytoseiidae Predatory Mites is occurred before ovum output, and even egg cell is just sent out
Educate front and rear.Therefore, it is to understand fully a pass of its reproduction mechanism that research is carried out to the growth course of Phytoseiidae Predatory Mites embryonated egg
Key point, to study the growth course of embryonated egg, it is necessary to by its reproductive system（The development structure of phytoseiid mite embryonated egg）Solution is cut
To be studied.
Electron microscopic observation is scanned to Predatory Mites reproductive system, can be from the structure group of ultra micro reproductive system from horizontal
Into, this transport pathway after can entering for its clear and definite male mite sperm in female mite body and with the binding site of egg cell provide ginseng
Examine.
But because the too small comparison of individual of Predatory Mites is fragile, is full of in vivo by opaque body fluid, it is difficult to its system genitale
System is accurately positioned, slightly accidentally may result in polypide and crush, and how available data is not dissected and how to be observed without related again
The report of its reproductive system, therefore electron microscopic observation Predatory Mites reproductive system how is obtained and is scanned, have to be solved.
The content of the invention
, can be with using this method it is an object of the invention to provide the method that Predatory Mites reproductive system is observed under ESEM
Predatory Mites reproductive system is separately separated out and ultra micro observation is carried out under ESEM.
The present invention is the female mite of Phytoseiidae Predatory Mites and male mite in first being mated, then is soaked by lactic acid, be dehydrated, do
Dry, viscous platform, metal spraying processing, the method that reproductive system is finally observed under ESEM, concrete operation step are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Predatory Mites raising population, choose ovum and individually raise extremely into mite
Male and female mite pairing is carried out afterwards, according to different needs by the male and female individual of the different time in mating process together with breeding apparatus one
Rise and be positioned over ultra low temperature freezer（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Predatory Mites male and female mite in mating, male and female mite is placed on slide with fine, soft fur pen, then
Male and female individual is separated with No. 0 insect needle and fine, soft fur pen, finally chosen male and female mite in the dactylethrae for filling lactic acid respectively, often
Temperature is placed 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
The present invention obtains Predatory Mites reproductive system first using very simple cleverly method, and realizes first in scanning electricity
The purpose of the trickle external structure of Microscopic observation reproductive system, the acquisition of reproductive system can provide to observe its structure under ESEM
Raw material, and the result of ESEM is advantageous to the research to reproductive system.
Embodiment
Embodiment is by taking Amblyseius persidolongispinosus as an example, the method for observation Predatory Mites reproductive system under ESEM
This method is the female mite of Amblyseius persidolongispinosus, male mite in first being mated, then is soaked by lactic acid, be dehydrated, be dry, be viscous
Platform, metal spraying processing, the method that Amblyseius persidolongispinosus reproductive system is finally observed under ESEM, concrete operation step are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Amblyseius persidolongispinosus raising population, choose ovum and individually raise
To into progress male and female mite pairing after mite, needed according to different by the male and female individual of the different time in mating process together with raising
Device is positioned over ultra low temperature freezer together（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Amblyseius persidolongispinosus male and female mite in mating, male and female mite is placed into slide with fine, soft fur pen
On, then with No. 0 insect needle and fine, soft fur pen male and female individual is separated, finally chosen male and female mite into the dactylethrae for filling lactic acid respectively
In, room temperature 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
Claims (1)
1. the method for Predatory Mites reproductive system is observed under ESEM, it is characterized in that the Phytoseiidae predation in first being mated
The female mite of mite, male mite, then soaked by lactic acid, be dehydrated, dry, viscous platform, metal spraying processing, finally observe system genitale under ESEM
The method of system, concrete operation step are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Predatory Mites raising population, choose ovum and individually raise extremely into mite
Male and female mite pairing is carried out afterwards, according to different needs by the male and female individual of the different time in mating process together with breeding apparatus one
Rise and be positioned over ultra low temperature freezer（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Predatory Mites male and female mite in mating, male and female mite is placed on slide with fine, soft fur pen, then
Male and female individual is separated with No. 0 insect needle and fine, soft fur pen, finally chosen male and female mite in the dactylethrae for filling lactic acid respectively, often
Temperature is placed 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
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| CN201710808937.5A CN107421793A (en) | 2017-09-09 | 2017-09-09 | The method that Predatory Mites reproductive system is observed under ESEM |
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| CN201710808937.5A CN107421793A (en) | 2017-09-09 | 2017-09-09 | The method that Predatory Mites reproductive system is observed under ESEM |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN104406834A (en) * | 2014-11-04 | 2015-03-11 | 中国水产科学研究院东海水产研究所 | Marine fish sperm staining method |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
-
2017
- 2017-09-09 CN CN201710808937.5A patent/CN107421793A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN104406834A (en) * | 2014-11-04 | 2015-03-11 | 中国水产科学研究院东海水产研究所 | Marine fish sperm staining method |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
Non-Patent Citations (2)
| Title |
|---|
| 吴桂华等: "《粉尘螨生殖***形态学研究》", 《昆虫学报》 * |
| 吴聪明: "《蠕形螨扫描电镜样本的制备方法》", 《动物学杂志》 * |
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Application publication date: 20171201 |