CN107568167A - Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method - Google Patents
Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method Download PDFInfo
- Publication number
- CN107568167A CN107568167A CN201710808914.4A CN201710808914A CN107568167A CN 107568167 A CN107568167 A CN 107568167A CN 201710808914 A CN201710808914 A CN 201710808914A CN 107568167 A CN107568167 A CN 107568167A
- Authority
- CN
- China
- Prior art keywords
- mite
- male
- spermatophore
- female
- mating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method, it is characterized in that first obtain the Phyloseiulus nersimilis hero mite it is readily seen that spermatophore, then the method for making and spermatophore being observed under ESEM, prepared and 5 parts of observation comprising the acquisition of male and female mite in mating, the sizing of male and female mite, the separating of male and female mite, the storage of male mite, spermatophore ESEM sample.The present invention is obtained with Phyloseiulus nersimilis hero mite spermatophore using very simple cleverly method, and the purpose that the trickle external structure of spermatophore is observed under ESEM can be realized, the acquisition of spermatophore can provide raw material to observe spermatophore structure under ESEM, and the research that the result of ESEM is advantageous to the other directions of spermatophore lays the first stone, provides enlightenment.
Description
Technical field
The present invention relates to Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method, belong to Phyloseiulus nersimilis biology
Basic research field.
Background technology
Phyloseiulus nersimilis is gonochorism animal, must can just be laid eggs by male and female mite mating after fertilization and breed offspring, after
In generation, no matter male and female were come from development of fertilized ova.
Available data mates for Phyloseiulus nersimilis male and female mite to be formed the understanding of embryonated egg and is：Male mite sperm is female in entrance
It is stored in before in mite body in spermatophore, spermatophore is filled in female mite copulatory opening by female mite（Bibliography：Liang Lairong, 1979, plant protected
How shield, mite class live）, sperm comes out from spermatophore slow release again, is transferred to spermatheca and ovum combines, form fertilization
Ovum.But how to enter in female mite body for Phyloseiulus nersimilis hero mite sperm and male mite sperm does not have with the presence or absence of dimorphism
Report, to research and solve this series of problems, it is necessary first to obtain spermatophore, observational study is carried out to spermatophore.
Due to very small only hundreds of microns of Phyloseiulus nersimilis individual, its spermatophore just goes out when there was only the mating of male and female mite
Existing, male and female mite being superimposed together closely in mating process, mating time is also very short only 1 hour or so, and spermatophore
Size only has more than ten micron and is hidden between the outside of belly of male and female mite body close contact, is intuitively hardly visible, existing money
The method for expecting also how not obtain spermatophore.Therefore think that acquisition spermatophore is very difficult, it is more tired that electron microscopic observation is scanned to spermatophore
Hardly possible is, it is necessary to open up a kind of method for obtaining Phyloseiulus nersimilis spermatophore and electron microscopic observation being scanned to spermatophore.
The content of the invention
It is an object of the invention to provide a kind of side for making and Phyloseiulus nersimilis hero mite spermatophore being observed under ESEM
Method, using this method can be trickleer observe Phyloseiulus nersimilis spermatophore external structure.
The present invention is to obtain the Phyloseiulus nersimilis hero mite it is readily seen that spermatophore, then makes and is observed under ESEM
The method of spermatophore, swept comprising the acquisition of male and female mite in mating, the sizing of male and female mite, the separating of male and female mite, the storage of male mite, spermatophore
Retouch electron microscope specimen and prepare and be with 5 parts of observation, concrete operations：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose small plant of Chile and pacify
Individually raising is into progress male and female mite pairing after mite for the ovum of mite, and 10 minutes or so male and female mites of pairing can mate, so as to be handed over
With middle male and female mite；
（2）The sizing of male and female mite in mating：Within Phyloseiulus nersimilis post-coitum 0-120 minutes, it will be in mating process
Male and female individual be positioned over 3-5 minutes in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carry out male and female mite and determine
Type；
（3）The separation of male and female mite after sizing：Male and female mite is taken out together with breeding apparatus after sizing, then under the microscope, used
Fine, soft fur pen chooses the male and female mite of mating on slide from breeding apparatus, recycles the instrument such as No. 0 insect needle and fine, soft fur pen will
The male and female mite of mating gently separates, and prevents from destroying spermatophore；
（4）The storage of male mite：After male and female mite successfully separates, male mite is chosen with fine, soft fur nib and is put into 70% alcohol in dactylethrae
Preserve；
（5）Spermatophore ESEM sample prepares and observation：Comprising dehydration, dry, 5 steps of viscous platform, metal spraying and observation, specifically
For：1）Dehydration：Gradient alcohol dehydration is carried out to the male mite being stored in dactylethrae, the concentration of alcohol is respectively：80%, 90%,
100%, first upper strata alcohol is suctioned out with rubber head dropper, then adds the alcohol of next gradient, so circulation is until 100%
Alcohol；2）Dry：Sample is placed on sample stage, is dried 2~3 hours using instrument is dried in vacuo；3）Viscous platform：In microscope
It is lower to be chosen male mite to viscous platform with fine, soft fur pen, make its outside of belly upward, the outside of belly is pressed lightly on insect needle, to ensure polypide with leading
Electric glue can be in close contact with excellent conductive performance；4）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；5）Electronic Speculum is seen
Examine：Spermatophore is observed under ESEM, spermatophore is located at below the spermatodactyl of male mite.
The present invention is obtained with Phyloseiulus nersimilis hero mite spermatophore using very simple cleverly method, and can realize
The purpose of the trickle external structure of spermatophore is observed under ESEM, the acquisition of spermatophore can provide to observe spermatophore structure under ESEM
Raw material, and the research that the result of ESEM is advantageous to the other directions of spermatophore lays the first stone, provides enlightenment.
Embodiment
Embodiment Phyloseiulus nersimilis spermatophore scanning electron microscopic observation method
This method is after artificially obtaining spermatophore from the Phyloseiulus nersimilis hero mite gnathosoma to mate, to be seen under ESEM
The method for examining spermatophore, include the acquisition of male and female mite, the sizing of male and female mite, the separation of male and female mite, the acquisition of spermatophore, scanning in mating
The part of electron microscopic observation 5, concrete operations are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose small plant of Chile and pacify
Individually raising is into progress male and female mite pairing after mite for the ovum of mite, and 10 minutes or so male and female mites of pairing can mate, so as to be handed over
With middle male and female mite；
（2）The sizing of male and female mite in mating：Within Phyloseiulus nersimilis post-coitum 0-120 minutes, it will be in mating process
Male and female individual be positioned over 3-5 minutes in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carry out male and female mite and determine
Type；（3）The separation of male and female mite after sizing：Male and female mite is taken out together with breeding apparatus after sizing, then under the microscope, with thin
Writing brush chooses the male and female mite of mating on slide from breeding apparatus, recycles the instrument such as No. 0 insect needle and fine, soft fur pen to hand over
The male and female mite matched somebody with somebody gently separates, and prevents from destroying spermatophore；
（4）The storage of male mite：After male and female mite successfully separates, male mite is chosen with fine, soft fur nib and is put into 70% alcohol in dactylethrae
Preserve；
（5）Spermatophore ESEM sample prepares and observation：Comprising dehydration, dry, 5 steps of viscous platform, metal spraying and observation, specifically
For：1）Dehydration：Gradient alcohol dehydration is carried out to the male mite being stored in dactylethrae, the concentration of alcohol is respectively：80%, 90%,
100%, first upper strata alcohol is suctioned out with rubber head dropper, then adds the alcohol of next gradient, so circulation is until 100%
Alcohol；2）Dry：Sample is placed on sample stage, is dried 2~3 hours using instrument is dried in vacuo；3）Viscous platform：In microscope
It is lower to be chosen male mite to viscous platform with fine, soft fur pen, make its outside of belly upward, the outside of belly is pressed lightly on insect needle, to ensure polypide with leading
Electric glue can be in close contact with excellent conductive performance；4）Metal spraying, it is gold-plated to mite whole body；5）Spermatophore is observed under ESEM,
Spermatophore is located at below the spermatodactyl of male mite.
Claims (1)
1. Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method, it is characterized in that Chilean small plant for first obtaining generation spermatophore is pacified
Mite hero mite, a series of processing are then carried out to male mite and are fabricated under ESEM the directly method of observation spermatophore, among these
Include the acquisition of male and female mite in mating, the sizing of male and female mite, the separating of male and female mite, the storage of male mite, spermatophore ESEM sample
Prepare and be with 5 parts of observation, concrete operations：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose small plant of Chile and pacify
Individually raising is into progress male and female mite pairing after mite for the ovum of mite, and 10 minutes or so male and female mites of pairing can mate, so as to be handed over
With middle male and female mite；
（2）The sizing of male and female mite in mating：Within Phyloseiulus nersimilis post-coitum 0-120 minutes, it will be in mating process
Male and female individual be positioned over 3-5 minutes in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carry out male and female mite and determine
Type；
（3）The separation of male and female mite after sizing：Male and female mite is taken out together with breeding apparatus after sizing, then under the microscope, used
Fine, soft fur pen chooses the male and female mite of mating on slide from breeding apparatus, recycles the instrument such as No. 0 insect needle and fine, soft fur pen will
The male and female mite of mating gently separates, and prevents from destroying spermatophore；
（4）The storage of male mite：After male and female mite successfully separates, male mite is chosen with fine, soft fur nib and is put into 70% alcohol in dactylethrae
Preserve；
（5）Spermatophore ESEM sample prepares and observation：Comprising dehydration, dry, 5 steps of viscous platform, metal spraying and observation, specifically
For：1）Dehydration：Gradient alcohol dehydration is carried out to the male mite being stored in dactylethrae, the concentration of alcohol is respectively：80%, 90%,
100%, first upper strata alcohol is suctioned out with rubber head dropper, then adds the alcohol of next gradient, so circulation is until 100%
Alcohol；2）Dry：Sample is placed on sample stage, is dried 2~3 hours using instrument is dried in vacuo；3）Viscous platform：In microscope
It is lower to be chosen male mite to viscous platform with fine, soft fur pen, make its outside of belly upward, the outside of belly is pressed lightly on insect needle, to ensure polypide with leading
Electric glue can be in close contact with excellent conductive performance；4）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；5）Electronic Speculum is seen
Examine：Spermatophore is observed under ESEM, spermatophore is located at below the spermatodactyl of male mite.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710808914.4A CN107568167A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710808914.4A CN107568167A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107568167A true CN107568167A (en) | 2018-01-12 |
Family
ID=61032713
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710808914.4A Pending CN107568167A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107568167A (en) |
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 CN201710808914.4A patent/CN107568167A/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: "《粉尘螨生殖系统形态学研究》", 《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》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tamura et al. | An improved DNA injection method for silkworm eggs drastically increases the efficiency of producing transgenic silkworms | |
| CN107568167A (en) | Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method | |
| CN107515314A (en) | A kind of method for obtaining and small new neoseiulus californicus spermatophore being observed under ESEM | |
| CN107655913A (en) | The acquisition of Tianjin river amblyseius as predatory mite spermatophore and the method observed under ESEM | |
| CN107576682A (en) | The method that Predatory Mites spermatophore is observed under ESEM | |
| Thompson et al. | Transmission and scanning electron microscopy of the accessory cells and chorion during development of Ciona intestinalis type B embryos and the impact of their removal on cell morphology | |
| CN107581156A (en) | A kind of acquisition methods of small new neoseiulus californicus spermatophore | |
| CN107568168A (en) | A kind of method for obtaining, preserving Tianjin river amblyseius as predatory mite spermatophore | |
| CN107632033A (en) | A kind of method for obtaining and small new neoseiulus californicus spermatophore being observed under transmission electron microscope | |
| CN107475176A (en) | A kind of method for obtaining Predatory Mites spermatophore | |
| Liu et al. | Morphology and husbandry of the large milkweed bug, Oncopeltus fasciatus | |
| CN107515315A (en) | Phyloseiulus nersimilis hero mite spermatophore transmission electron microscope observing method | |
| CN107478669A (en) | The method that Predatory Mites spermatophore is observed under transmission electron microscope | |
| CN107515145A (en) | A kind of method for making small new neoseiulus californicus sperm slide sample | |
| CN107576542A (en) | A kind of acquisition Predatory Mites hero mite sperm and the method for making slide sample | |
| Neznanova | Comparative analysis of gamete ultrastructure in big-scaled redfin Tribolodon hakonensis (Cyprinidae) from Southern Primorye and Sakhalin | |
| Meyer et al. | An electron microscope study of embryonic heart muscle cells grown in tissue cultures | |
| Alberti | On predation in Epicriidae (Gamasida, Anactinotrichida) and fine-structural details of their forelegs | |
| CN107475177A (en) | The preparation method of Phyloseiulus nersimilis spermatophore | |
| CN107655733A (en) | The acquisition of Phyloseiulus nersimilis sperm and slide sample preparation method | |
| Liu et al. | Ultra-and microstructure of the female reproductive system of Matsucoccus matsumurae | |
| CN107632031A (en) | The acquisition of Tianjin river amblyseius as predatory mite spermatophore and its observational technique under transmission electron microscope | |
| CN107568199A (en) | The preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample | |
| CN107589004A (en) | A kind of method for making and obtaining Tianjin river amblyseius as predatory mite reproductive system | |
| Usmani | Biological investigations on some species of Anagrus (Hymenoptera, Mymaridae), egg parasitoids of leafhoppers (Hemiptera) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180112 |