CN111044546A - Parasitic bee electron microscope method, parasitic bee electron microscope system and storage medium - Google Patents
Parasitic bee electron microscope method, parasitic bee electron microscope system and storage medium Download PDFInfo
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- CN111044546A CN111044546A CN201911254853.7A CN201911254853A CN111044546A CN 111044546 A CN111044546 A CN 111044546A CN 201911254853 A CN201911254853 A CN 201911254853A CN 111044546 A CN111044546 A CN 111044546A
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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
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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
- G01N23/225—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 using electron or ion
- G01N23/2251—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 using electron or ion using incident electron beams, e.g. scanning electron microscopy [SEM]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
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Abstract
The invention discloses a parasitic bee electron microscope method, a parasitic bee electron microscope system and a storage medium, wherein the method comprises the following steps: obtaining a parasitic bee insect sample and a parasitic bee antenna; processing the parasitic bee antenna by an ethanol solution to obtain a parasitic bee antenna sample; trimming the antenna sample, and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape; treating the parasitic bee antenna sample by using a discharge prevention liquid; carrying out surface gold plating treatment on the antenna sample; and acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope. Aiming at the unique characteristics of parasitic wasp antennae, the invention configures electron microscope voltage for an electron microscope, uses ethanol solution and discharge prevention liquid to treat an antenna sample, and provides an effective method for researching the antenna receptor ultrastructure of parasitic wasps by the electron microscope. The invention can be widely applied to the field of biological electron microscopes.
Description
Technical Field
The invention relates to the field of biological electron microscopes, in particular to a parasitic bee electron microscope method, a parasitic bee electron microscope system and a storage medium.
Background
Periplaneta americana has existed on the earth for over 2 hundred million years, adapts to various environmental conditions, and becomes a pest which can carry various viruses and pollute the living environment of human beings. At present, the control of the periplaneta americana mainly depends on chemical pesticides and bait agents. The periplaneta americana population, although controlled to some extent, thereby increases the risk of developing drug resistance to periplaneta americana, and in addition toxic chemical materials are transmitted to other animals through the food chain, causing greater environmental hazards.
Aiming at the problems, the control of the periplaneta americana by using parasitic wasps becomes a new direction for research of scientists in various countries. Currently 9 blattaria egg parasitic wasps have been reported, but there is a systematic study of their parasitic life history. The Chouioia haiwensis is mainly studied, and the Chouioia haiwensis is proved to be capable of effectively controlling the population density of the periplaneta americana. Successful use of parasitic wasps in biological control depends on their ability to host in complex chemical and physical environments. Research shows that after parasitic wasps with learning experience are released in the field, the hosts can be found more accurately and quickly through the odor, and the method has higher search efficiency and parasitic rate on the hosts.
The olfactory sensor plays an important role in a series of behaviors such as parasitic wasps searching for hosts and determining whether to lay eggs on the hosts. Insect sensates are distributed in the antennal, plantar, abdominal, ovipositor, etc., and have different functions and forms. At present, the ultrastructure of a receptor on an antenna of other insects is researched in China, but the research on the ultrastructure of cockroach egg parasitic wasps such as Chouioia haifanensis and the antenna of the Qibellado wasp is blank. Considering that the cockroach egg parasitic wasp has great significance for controlling the American cockroach, and the antenna sensor has a very important role in seeking a host, research on the antenna sensor of the parasitic wasp is urgently needed. But nowadays, an effective electron microscope method for researching the ultrastructure of parasitic wasps is lacked.
Disclosure of Invention
In order to solve the above technical problems, an object of the present invention is to provide an effective parasitic bee-wave microscopy method, system and storage medium.
The invention provides a parasitic bee electron microscope method, which comprises the following steps:
obtaining a parasitic bee insect sample and a parasitic bee antenna;
processing the parasitic bee antenna by an ethanol solution to obtain a parasitic bee antenna sample;
trimming the antenna sample, and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape;
treating the parasitic bee antenna sample by using a discharge prevention liquid;
carrying out surface gold plating treatment on the antenna sample;
and acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope.
Further, the step of obtaining the parasitic wasp insect body sample and the parasitic wasp antenna comprises the following steps:
obtaining a plurality of female parasitic wasps and a plurality of male parasitic wasps which are not mated from a culture place according to the genital shapes of the parasitic wasps;
placing the parasitic wasps in a centrifuge tube or a culture dish;
processing parasitic wasps in the culture dish by carbon dioxide to obtain parasitic wasp insect body samples;
and (3) processing the parasitic wasp body sample to obtain the parasitic wasp antenna.
Further, the step of processing the parasitic bee antenna by the ethanol solution to obtain a parasitic bee antenna sample comprises the following steps:
obtaining the antenna of the parasitic wasp, and placing the antenna in 30% ethanol solution for treatment;
carrying out ultrasonic cleaning on the antenna treated by the ethanol solution;
gradually dehydrating the tentacles by using 50%, 70%, 80%, 90% and 100% ethanol solutions respectively, wherein each dehydration is carried out for 15 minutes;
placing the antenna at a ventilated place for airing to obtain an antenna sample;
further, the method also comprises a step of storing the antenna sample, and the step specifically comprises the following steps:
placing an antenna sample in a centrifugal tube, wherein a 10% ethanol solution is stored in the centrifugal tube;
and (4) storing the centrifuge tube in an environment at-80 ℃.
Further, the step of performing surface gold plating treatment on the antenna sample comprises the following steps:
and carrying out a plurality of gold-plating operations on the antenna sample by using an ion sputtering instrument, wherein the duration of one gold-plating operation is 30 seconds.
Further, the step of processing the parasitic bee antenna sample by using the discharge prevention liquid comprises the following steps:
and after the antenna sample is fixed on the sample table, dropwise adding a discharge prevention liquid onto the antenna sample.
The invention also provides a parasitic bee electron microscope system, which comprises:
the parasitic wasp body sample acquisition module is used for acquiring a parasitic wasp body sample and a parasitic wasp antenna;
the parasitic bee antenna sample acquisition module is used for processing the parasitic bee antenna through an ethanol solution to acquire a parasitic bee antenna sample;
the antenna sample fixing module is used for trimming the antenna sample and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape;
the discharging prevention liquid module is used for processing the parasitic bee antenna sample by using the discharging prevention liquid;
the gold plating processing module is used for carrying out surface gold plating processing on the antenna sample;
and the electron microscope image acquisition module is used for acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope.
The invention also provides a parasitic bee electron microscope system, which comprises:
at least one processor;
at least one memory for storing at least one program;
when executed by the at least one processor, cause the at least one processor to implement a parasitic bee-wave microscopy method as described above.
The present invention also proposes a storage medium having stored therein processor-executable instructions for performing a parasitic bee-mirror method as described above when executed by a processor.
One or more of the above-described embodiments of the present invention have the following advantages: aiming at the unique characteristics of parasitic wasp antennae, the invention configures proper electron microscope voltage for an electron microscope, uses ethanol solution and discharge prevention liquid to treat an antenna sample, and provides an effective method for researching the antenna receptor ultrastructure of parasitic wasps by the electron microscope.
Drawings
FIG. 1 is a flow chart of a parasitic galvano-mirror method of the present invention;
fig. 2 is a schematic structural diagram of a parasitic bee electron microscope system according to the present invention.
Detailed Description
The technical scheme of the invention is described in detail in the following with reference to the accompanying drawings.
Referring to fig. 1, a parasitic bee electric microscopy method includes the following steps:
s1 obtaining a parasitic wasp body sample and a parasitic wasp antenna.
And S2, processing the parasitic bee antenna through an ethanol solution to obtain a parasitic bee antenna sample.
S3, trimming the antenna sample, and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape.
S4 processes the parasitic bee antenna sample using the discharge prevention liquid.
S5, carrying out surface gold plating treatment on the antenna sample.
S6, acquiring the electron microscope image of the antenna by configuring the voltage of the electron microscope.
The detailed steps of the method are described in connection with one embodiment of the method.
Taking the Chouioia cunea Yang in parasitic wasps as an example, collecting the Chouioia cunea Yang which is propagated and bred progressively in a Periplaneta americana breeding room at the stage of obtaining a Chouioia cunea body sample, wherein the breeding conditions of the parasitic wasps are that a gauze special cube-shaped breeding box (side length: 40cm) and the temperature is (2 ((1) (DEG C., light period: 14:10(L: D) (a small amount of Periplaneta americana pods and 10% honey water are placed in a gauze).
Generally, Chouioia hampsoia which just emerges from the feather pod is taken, and 5 female insects and 5 male insects are taken according to the genital morphology of the Chouioia hamoia. The bodies were placed in 2.0ml centrifuge tubes and smoked with carbon dioxide for use.
After obtaining the Chouioia hainanensis body sample, placing the male and female Chouioia hainanensis fumigated by carbon dioxide on a dissecting table, shearing the head of the Chouioia hainanensis by using a dissecting scalpel under a dissecting mirror, taking down the tentacles from the head, and quickly placing the Chouioia hainanensis body sample in 30% ethanol solution. And then ultrasonically cleaning the soaked tentacles for 100-120 seconds, gradually dehydrating the soaked tentacles for 15 minutes each time for 3 times by using ethanol solutions with different concentrations (50%, (0%, 80%, 90% and 100%), and finally placing the tentacles in a ventilation position to naturally dry the tentacles so as to finally obtain the tentacles of Chouioia hamii.
Then using tweezers to pick the tentacle under a dissecting mirror, trimming the tentacle by using a dissecting scalpel, adhering the tentacle to a sample table by using conductive adhesive on the back, the front and the two side surfaces, and using the conductive adhesive to observe the whole survival state of the tentacle, a thorn-shaped sensor at the base of a handle section and a peduncle section and a sensor on the surface of a small link, a cord section and a bar section. And finally, adhering and fixing the antenna sample of the Chouioia hampsii on a sample table by using conductive adhesive tape. Besides the four mentioned observation surfaces, it is also necessary to cut and erect a plurality of subsections divided by the antenna cord section, and observe the distribution rule of various sensors on the subsections from the cross section direction. It is also noted that all of the above mentioned viewing surfaces must be kept clean and free of contamination and damage.
After the fixing treatment of the Hashima tentacle antenna sample is completed, the sample is subjected to gold plating operation by using an ion sputtering instrument, and the gold plating time is 30 seconds at a time. Because the antenna is divided into a plurality of sections, large gaps are formed at the joints of the sections, the metal conducting layer is easy to be discontinuous, and a local charging phenomenon is formed, so that an image is unclear. Therefore, to prevent the discontinuity of the conductive layer, gold should be plated at least 4 times.
If the local unclear phenomenon still occurs in the observation antenna, a special discharge prevention liquid for a scanning electron microscope is used to solve the problem of discontinuous metal coating on the surface of the antenna. After the antenna sample is fixed, a drop of discharge prevention liquid is firstly dripped, and after the liquid is naturally dried, gold plating is carried out; or when the observation image shows that the local part is unclear, the sample is taken out and dripped with the discharge prevention liquid, and then the sample is observed on the machine, so that the charging effect caused by poor conduction can be effectively prevented.
The treated antenna sample of the Chouioia hampsonii is observed by adopting a KYKY-1000B scanning electron microscope. Because a large number of hair-type receptors exist on the antenna and a remarkable tip effect exists, attention is paid to the adoption of a proper electron microscope accelerating voltage, the voltage cannot be too high, and a voltage with the voltage of 5-12 kV (10 kV is selected in general) is recommended. Also, it is noted that the perceived length and diameter of the observation must be measured to ensure that the object is in the plane of the field of view, making an unsuitable angle can result in inaccurate early measurement data.
The detailed steps of the method are described in connection with another embodiment of the method.
For another parasitic wasp, the sailnet wasp, 5 of the unmatched female and male swarms of the sailnet wasp were also selected from the rearing population, similar to the procedure for selecting Chouioia haichii. After obtaining the body of the Qibellied Apis cerana, the obtained body should be placed in a culture dish with the diameter of 12cm, and the body is fumigated by carbon dioxide for later use.
The remaining subsequent processing steps for obtaining the antenna sample of the sailflower bramble are the same as those of the Chouioia hampsii.
Referring to fig. 2, the present invention further provides a parasitic bee electric mirror system, including:
the parasitic wasp body sample acquisition module is used for acquiring a parasitic wasp body sample and a parasitic wasp antenna;
the parasitic bee antenna sample acquisition module is used for processing the parasitic bee antenna through an ethanol solution to acquire a parasitic bee antenna sample;
the antenna sample fixing module is used for trimming the antenna sample and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape;
the discharging prevention liquid module is used for processing the parasitic bee antenna sample by using the discharging prevention liquid;
the gold plating processing module is used for carrying out surface gold plating processing on the antenna sample;
and the electron microscope image acquisition module is used for acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope.
The invention also provides a parasitic bee electron microscope system, which comprises:
at least one processor;
at least one memory for storing at least one program;
when executed by the at least one processor, cause the at least one processor to implement a parasitic bee-wave microscopy method as described above.
The present invention also proposes a storage medium having stored therein processor-executable instructions for performing a parasitic bee-mirror method as described above when executed by a processor.
In summary, compared with the prior art, the invention has the following advantages:
(1) aiming at the unique characteristics of parasitic wasp antennae, the invention configures proper electron microscope voltage for an electron microscope, uses ethanol solution and discharge prevention liquid to treat an antenna sample, and provides an effective method for researching the antenna receptor ultrastructure of parasitic wasps by the electron microscope.
(2) Aiming at the problem of discontinuous gold plating on the surface of the antenna, the invention adopts a method of plating gold for 30 seconds for a plurality of times and uses a discharge prevention liquid to treat an antenna sample, thereby effectively preventing the problem caused by the local charging phenomenon and leading the observation image of the antenna to be clear.
The step numbers in the above method embodiments are set for convenience of illustration only, the order between the steps is not limited at all, and the execution order of each step in the embodiments can be adapted according to the understanding of those skilled in the art.
While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (9)
1. A parasitic bee electron microscope method is characterized by comprising the following steps:
obtaining a parasitic bee insect sample and a parasitic bee antenna;
processing the parasitic bee antenna by an ethanol solution to obtain a parasitic bee antenna sample;
trimming the antenna sample, and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape;
treating the parasitic bee antenna sample by using a discharge prevention liquid;
carrying out surface gold plating treatment on the antenna sample;
and acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope.
2. The parasitic bee electron microscopy method according to claim 1, characterized in that: the step of obtaining the parasitic bee body sample and the parasitic bee antenna comprises the following steps:
obtaining a plurality of female parasitic wasps and a plurality of male parasitic wasps which are not mated from a culture place according to the genital shapes of the parasitic wasps;
placing the parasitic wasps in a centrifuge tube or a culture dish;
processing parasitic wasps in the culture dish by carbon dioxide to obtain parasitic wasp insect body samples;
and (3) processing the parasitic wasp body sample to obtain the parasitic wasp antenna.
3. The parasitic bee electron microscopy method according to claim 1, characterized in that: the step of processing the parasitic bee antenna by the ethanol solution to obtain the parasitic bee antenna sample comprises the following steps:
obtaining the antenna of the parasitic wasp, and placing the antenna in 30% ethanol solution for treatment;
carrying out ultrasonic cleaning on the antenna treated by the ethanol solution;
gradually dehydrating the tentacles by using 50%, 70%, 80%, 90% and 100% ethanol solutions respectively, wherein each dehydration is carried out for 15 minutes;
and placing the antenna in a ventilated place for airing to obtain an antenna sample.
4. The parasitic bee electron microscopy method according to claim 1, characterized in that: the method also comprises a step of storing the antenna sample, and the step specifically comprises the following steps:
placing an antenna sample in a centrifugal tube, wherein a 10% ethanol solution is stored in the centrifugal tube;
and (4) storing the centrifuge tube in an environment at-80 ℃.
5. The parasitic bee electron microscopy method according to claim 1, characterized in that: the step of performing surface gold plating treatment on the antenna sample comprises the following steps:
and carrying out a plurality of gold-plating operations on the antenna sample by using an ion sputtering instrument, wherein the duration of one gold-plating operation is 30 seconds.
6. The parasitic bee electron microscopy method according to claim 1, characterized in that: the step of processing the parasitic bee antenna sample by using the discharge prevention liquid comprises the following steps:
and after the antenna sample is fixed on the sample table, dropwise adding a discharge prevention liquid onto the antenna sample.
7. A parasitic bee electron microscope system, comprising:
the parasitic wasp body sample acquisition module is used for acquiring a parasitic wasp body sample and a parasitic wasp antenna;
the parasitic bee antenna sample acquisition module is used for processing the parasitic bee antenna through an ethanol solution to acquire a parasitic bee antenna sample;
the antenna sample fixing module is used for trimming the antenna sample and fixing the trimmed antenna sample on a sample table through a conductive adhesive tape;
the discharging prevention liquid module is used for processing the parasitic bee antenna sample by using the discharging prevention liquid;
the gold plating processing module is used for carrying out surface gold plating processing on the antenna sample;
and the electron microscope image acquisition module is used for acquiring an electron microscope image of the antenna by configuring the voltage of the electron microscope.
8. A parasitic bee electron microscope system, comprising:
at least one processor;
at least one memory for storing at least one program;
when executed by the at least one processor, cause the at least one processor to implement a parasitic bee-electronic microscopy method as defined in any one of claims 1-6.
9. A storage medium having stored therein processor-executable instructions, which when executed by a processor, are configured to perform a parasitic bee-shine mirror method as claimed in any one of claims 1 to 6.
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| CN201911254853.7A CN111044546A (en) | 2019-12-10 | 2019-12-10 | Parasitic bee electron microscope method, parasitic bee electron microscope system and storage medium |
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| CN201911254853.7A CN111044546A (en) | 2019-12-10 | 2019-12-10 | Parasitic bee electron microscope method, parasitic bee electron microscope system and storage medium |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN109239112A (en) * | 2018-08-07 | 2019-01-18 | 重庆师范大学 | A kind of sample preparation methods for electron-microscope scanning |
-
2019
- 2019-12-10 CN CN201911254853.7A patent/CN111044546A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN109239112A (en) * | 2018-08-07 | 2019-01-18 | 重庆师范大学 | A kind of sample preparation methods for electron-microscope scanning |
Non-Patent Citations (3)
| Title |
|---|
| 刘玉双 等: "红缘吉丁（鞘翅目： 吉丁虫科）触角感器的扫描电镜观察", 《昆虫学报》 * |
| 洪健 等: "寄生蜂触角的扫描电镜样品制备技术", 《电子显微学报》 * |
| 车江萍 等: "二疣犀甲成虫头部感受器超微结构观察", 《热带作物学报》 * |
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Application publication date: 20200421 |