CN110286016B - Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof - Google Patents
Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof Download PDFInfo
- Publication number
- CN110286016B CN110286016B CN201910625649.5A CN201910625649A CN110286016B CN 110286016 B CN110286016 B CN 110286016B CN 201910625649 A CN201910625649 A CN 201910625649A CN 110286016 B CN110286016 B CN 110286016B
- Authority
- CN
- China
- Prior art keywords
- insect
- antenna
- pad table
- fixing
- irregular
- 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.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/2806—Means for preparing replicas of specimens, e.g. for microscopal analysis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Catching Or Destruction (AREA)
Abstract
The invention discloses a directional fixing device for irregular feeler of living insects and a manufacturing method thereof, belonging to the technical field of experimental sample treatment. The device includes slide glass, antenna pad platform, worm body pad platform, water storage little well device, ultraviolet glue, the fixed middle part that sets up at the slide glass of water storage little well device, antenna pad platform sets up on the slide glass and antenna pad platform is located water storage little well device, worm body pad platform sets up on the slide glass and worm body pad platform is located water storage little well device, worm body pad platform is located antenna pad platform side and contacts with antenna pad platform side, worm body pad bench upper surface sets up the viscose layer, the junction of antenna pad platform and worm body pad platform and antenna pad bench top coating are provided with ultraviolet glue. The invention realizes the directional fixation of the irregular antenna by utilizing the integration of a plurality of devices, is convenient for the observation, the positioning and the recording of a single sensor, has high experimental efficiency and low cost, and can be reused.
Description
Technical Field
The invention relates to a directional fixing device for a living body fruit fly insect with irregular feeler and a manufacturing method thereof, belonging to the technical field of experimental sample treatment.
Background
Feelers are important sensory organs of insects. The surface of the insect antenna is densely distributed with various types of micron-sized sensors, and a single sensor is a basic structural unit of an insect antenna sensing organ, is an information receiving device for sensing the external environment of an insect organism and carrying out communication, and can sense physical mechanical or chemical stimulation. In the process of searching for spouse, finding a host or a hunting habitat, insects are mainly completed through feeler sensing of sex pheromones, host plant volatile matters, sound, air and other chemical and physical stimuli, and the insects play important roles in the processes of insect foraging, host identification, spawning, mating, enemy avoidance and other behaviors. For example, cabbage butterflies (Pieris rapae) are sensitive to air-mediated mustard odor emanating from crucifers. Many insects can be attracted by the sweet and sour wine flavor emitted by damaged fruits. Sex hormones secreted by females of many moths can attract males outside kilometers to fly for mating. Therefore, the insect antenna can be utilized to sense the smell of chemical substances, dynamically measure and report the pest population, and trap, prevent and control pests and the like.
The feeler sensor has sensory receptor neurons with different functions distributed inside. Taking an odor sensor as an example, external odor substances pass through nanoscale holes on the surface of the sensor, finally reach olfactory receptor neurons through a series of recognition and transportation processes, finally, external chemical signals are converted into electric signals, and reach the central nervous system of the brain of the insect, so that insect behavioral reactions are caused. The study of electrophysiological response of a certain type of single sensor of the feeler organ of the insect has great significance for understanding the information coding rule and ecological behaviors of the insect on chemical smell and mechanical stimulus from a deeper microscopic level.
At present, the fixing technology of the antenna with a more regular shape (the antenna of the pattern insect drosophila is flat) generally adopts a gun head of a microscale sampler to fix the insect body, and then uses the tip of a glass electrode to press the back side of the flat antenna on the cover glass for fixing. Therefore, only the sensor in the back side direction can be exposed and recorded, and the single-sensor experiment in other directions cannot be realized; the fixing method for very small insect (mosquito) antenna is simpler, and the insect body and the antenna are directly adhered to the double faced adhesive tape and pressed on the double faced adhesive tape by the hair.
However, some insects, such as the family of faiidae insects, have larger insect bodies and antenna sizes, thicker cuticle and lower light transmission; the antenna sections are irregular in shape, and most of the antenna sections are elliptical; the thickness of each section of the antenna is different, and the height is different; the length of the sensor on the antenna is 5-20 micrometers; many types of sensors tend to be densely distributed across, and the distance between individual sensors is only 2-3 microns, making identification and localization difficult. In order to study the electrophysiological response of different types of sensors on the antenna of the insect of the family Bactrocera to different physical and chemical stimuli, it is necessary to use living insects, to resolve and locate the sensors at least at 1000 magnification, and to insert the recording electrode with a tip of 1 micron precisely into a single sensor base of a specific type (single sensor recording experiment). Also, the antenna cannot move slightly during recording, otherwise the electrode displacement will cause the target electrical signal to be lost. Therefore, it is necessary to fix the living insect and the antenna at the same time. More importantly, considering that the insect antenna has the back side, the abdomen side, the outside and the inside, the spatial distribution patterns of the sensors are different, the functions are different, and in order to fully understand the working mode of the antenna, the single sensor record of each angle needs to be realized, and the orientation of a certain angle needs to be fixed for the irregular antenna of the living insect shape. In addition, in order to distinguish and accurately position the single sensor, a multiplied by 100 water mirror is used under the condition of water to observe various types of single sensors, and micro electrodes such as tungsten wires are inserted into the base part of a certain type of sensors through microscopic operation, so that the insect body and the antenna are fixed, and meanwhile, the directional fixing device for the irregular antenna of the insect shape is required to be specially designed.
Disclosure of Invention
The invention aims to provide a living insect irregular antenna directional fixing device and a manufacturing method thereof, which can realize the rapid and synchronous fixation of insect bodies and extremely irregular antenna in shape, especially realize the directional fixation of the irregular antenna at a specific angle according to the position requirement of a single sensor experiment, meet the single sensor experiment micromanipulation with higher difficulty level, and solve the problems of specific angle fixation, thicker antenna stratum corneum, low transmittance, easy target movement caused by recording electrode puncture, single sensor spacing of only 2-3 microns and difficult accurate positioning operation.
The invention realizes the directional fixation of the irregular antenna by utilizing the integration of a plurality of devices, is convenient for the observation, the positioning and the recording of a single sensor, has high experimental efficiency and low cost, and can be reused.
The utility model provides an directional fixing device of irregular antenna of insect shape, including slide glass 1, antenna pad platform 6, worm body pad platform 3, water storage little well device 2, ultraviolet glue 5, water storage little well device 2 is fixed to be set up at the middle part of slide glass 1, antenna pad platform 6 sets up on slide glass 1 and antenna pad platform 6 is located water storage little well device 2, worm body pad platform 3 sets up on slide glass 1 and worm body pad platform 3 is located water storage little well device 2, worm body pad platform 3 is located antenna pad platform 6 side and contacts with antenna pad platform 6 side, worm body pad platform 3 upper surface sets up viscose layer 7, the junction of antenna pad platform 6 and worm body pad platform 3 and the coating of antenna pad platform 6 are provided with ultraviolet glue 5.
The water storage small well device 2 is in a circular column or a hollow polygonal column.
The distance between one end of the adhesive layer 7 near the antenna pad stand and the antenna pad stand 6 is less than 1/2 of the length of the insect head, and the width of the adhesive layer 7 is consistent with the width of the insect body pad stand 3. The height of the water storage small well device 2 is 2-3 mm.
The width of the ultraviolet glue 5 is larger than the width of the insect head.
Preferably, the insect body fixing device also comprises an insect body fixing line 4, wherein one end of the insect body fixing line 4 is fixedly arranged on the adhesive layer 7 of the insect body pad table 3; the insect fixing line 4 can better fix the insect body and avoid the struggling displacement of living insects.
Preferably, the number of the insect body fixing lines 4 is two, and the insect body fixing lines are respectively positioned at the head-chest section and the chest-abdomen section of the insect.
Preferably, the length of the antenna pad table 6 is 5-20 mm, the width is 3-5 mm, and the height is 1-2 mm; the length of the insect pad table 3 is 10-25 mm, the width is 8-20 mm, and the height is 0.5-1 mm.
Preferably, the material of the small well device is hot melt adhesive.
Preferably, the antenna pad stand 6 is a glass slide, and the worm pad stand 3 is a cover glass.
Preferably, the adhesive layer 7 is double-sided adhesive;
the antenna pad table can provide a base for directional fixation of the antenna, so that the height difference between the antenna and the insect body is convenient to coordinate, and the light transmittance is met; the insect body pad table is used for placing insect bodies and filling the height difference between the bodies and the antenna; the adhesive layer (double-sided adhesive) on the insect body pad table can reduce struggling of insect bodies, the insect body fixing line is arranged on the insect body pad table, the end of the insect body fixing line is fixedly adhered to the adhesive layer (double-sided adhesive), and the insect body fixing line is respectively pressed at the head-chest section and the chest-abdomen section to further fix the insect bodies; the connection part of the antenna pad table and the insect pad table and the ultraviolet adhesive on the antenna pad table can rapidly orient and fix the antenna and the head of the living insect without damaging the antenna structure and the function; the small well device is a water storage medium, can ensure that the type of the antenna sensor is determined under the magnification of 1000 times by means of a water objective lens (multiplied by 100), is positioned accurately, and is inserted into a specific sensor accurately by applying a micromanipulation technology.
The manufacturing method of the living insect irregular antenna directional fixing device comprises the following specific steps:
(1) Cutting the glass slide into an antenna pad table, and bonding the bottom surface of the antenna pad table to the middle part of another clean glass slide;
(2) Cutting the cover glass into an insect pad table, adhering the bottom surface of the insect pad table to the clean glass slide, and enabling the side end of the insect pad table to be in contact with the side surface of the antenna pad table;
(3) The adhesive layer is adhered and covered on the insect pad table;
(4) The ultraviolet glue is coated and arranged at the joint of the antenna pad table and the insect body pad table and on the antenna pad table, and the width of the ultraviolet glue is larger than the width of the insect body head;
(5) And a water storage small well device is arranged along the periphery of the antenna pad table and the insect pad table by adopting hot melt adhesive.
Further, the step (3) further comprises the step of adhering the insect body fixing line to the adhesive layer of the insect body pad table, and enabling the insect body fixing line to be respectively positioned at the head-chest section and the chest-abdomen section of the insect.
The using process of the directional fixing device for the irregular feeler of the shape of the living insect comprises the following steps:
in use, insect samples are prepared and treated: cutting off feet and wings of insects, then smearing a small amount of viscous ultraviolet glue on the joint of the antenna pad table and the insect body pad table and on the antenna pad table, and placing the abdomen sides of insect heads, breasts and abdomen on the adhesive layer (double-sided glue) on the insect body pad table to finish preliminary fixation of insect bodies; then, the abdominal part of the insect head is immersed in ultraviolet glue on the double-sided adhesive of the insect body pad table; under a split microscope, the antenna with a determined position and angle is initially fixed according to the antenna position to be positioned and observed, such as the abdomen side, the back side, the inner side or the outer side, by utilizing the characteristic of high viscosity of ultraviolet glue;
taking the antennal ventral fixation (dorsal exposure) as an example, firstly cutting off the feet and wings of insects, then smearing a small amount of viscous ultraviolet glue on the joint of the antennal pad table and the insect body pad table and on the antennal pad table, and placing the ventral sides of living insect heads, chest and abdomen with the feet and wings cut off on the adhesive layer (double-sided glue) on the insect body pad table to finish the preliminary fixation of the insect body; then, the abdominal part of the insect head is immersed in ultraviolet glue on the double-sided adhesive of the insect body pad table; under a split microscope, placing the abdomen side or the inclined abdomen side (back side is exposed) of the antenna into a small amount of ultraviolet glue by using a thin tweezer for preliminary fixation; solidifying the ultraviolet glue by an ultraviolet lamp, namely solidifying the ultraviolet glue by ultraviolet gel after irradiating the ultraviolet glue for 5-8 seconds by the ultraviolet lamp, and permanently fixing the antenna; in order to further fix the insect body, the insect body fixing line can be respectively arranged between the head and the chest and between the chest and the abdomen, and is tensioned and bonded with the adhesive layer (double-sided adhesive), so as to fix the insect body; finally, filling water into the small well device, and observing the antenna single sensor in the back side direction of the insect under the magnification of a high-power microscope and performing subsequent experimental operation.
Working principle:
the antenna pad table is used as a base for directional fixation of the antenna, so that the height difference between the antenna and the insect body is conveniently coordinated, and the light transmittance is met; placing the insect body by using an insect body pad table, filling the height difference between the body and the antenna, and primarily fixing the insect body; the adhesive layer is used for reducing struggle of insect bodies, so that the fixing is convenient; in order to further fix the insect bodies and prevent the insect bodies from struggling and deforming, an insect body fixing line is arranged on the insect body pad table; ultraviolet glue smeared on the joint of the antenna pad table and the insect body pad table is convenient for fixing the antenna and the head of the living insect; the ultraviolet glue is nontoxic and odorless and has toughness, and the ultraviolet glue can not interfere an insect olfactory sensor; the small well device is a water storage medium, so that the type of the sensor can be conveniently determined by means of a water mirror under 1000 times of amplification, and an electrode is inserted into the base part of the single sensor;
some insects such as the faithful fly family insects have larger individuals and are not easy to fix; compared with the characteristics of thinner horn layers of the antennas of the Drosophila, flat and easily fixed morphology, the horn layers of the antennas of the Drosophila are thicker (difficult to transmit light and difficult to insert electrodes), the morphology is very irregular, multiple types of sensors are densely and crosswise distributed, the distance is only 2-3 microns, the fixation difficulty is high, and the type identification, positioning and operation difficulties of the sensors are greatly exceeded that of the Drosophila.
The invention has the beneficial effects that:
(1) For some insects, the individuals of the faithful fly insects are large, and the insects are not easy to fix; compared with the characteristics of thinner horn layers of the antennas of the Drosophila, flat and easily fixed morphology, the horn layers of the antennas of the Drosophila are thicker (difficult to transmit light and difficult to insert electrodes), have irregular morphology, are densely and crosswise distributed with a plurality of types of sensors, have a spacing of only 2-3 microns, are difficult to fix, and are greatly more difficult to identify, position and operate than the Drosophila; the directional fixing device for the living insect irregular antenna can realize the rapid and synchronous fixation of the insect body and the extremely irregular antenna, especially can realize the directional fixation of the irregular antenna at a specific angle according to the position requirement of a single sensor experiment, meets the single sensor experiment microscopic operation of higher difficulty level, and solves the problems of fixed specific angle, thicker antenna stratum corneum, low transmittance, easy target movement caused by recording electrode puncture, and difficult accurate positioning operation, wherein the single sensor distance is only 2-3 microns;
(2) The directional fixing device for the irregular feeler of the living insect shape realizes directional fixing of all angles of the irregular feeler of the insect shape, and the living insect and the feeler are fixed at the same time and cannot displace, so that the single sensor recording signals of all angles are stably realized;
(3) The directional fixing device for the living insect feeler with irregular shape is suitable for fixing all living insect feelers of the Bactrocera genus; the fixing device is also suitable for fixing the feeler of insects in other families (including the Drosophila mode);
(4) The directional fixing device for the living insect irregular antenna is simple and convenient to operate, high in reliability, high in experimental efficiency, low in cost and capable of being reused.
Drawings
Fig. 1 is a perspective view of an orientation fixture of an irregularly shaped antenna of a living insect;
fig. 2 is a front view of an orientation fixture of an irregularly shaped antenna of a living insect;
in the figure, a glass slide, a 2-water storage small well device, a 3-insect body pad table, a 4-insect body fixing line, a 5-ultraviolet adhesive, a 6-antenna pad table and a 7-adhesive layer are arranged;
fig. 3 is an effect diagram of fixing the antenna at different angles in example 2, in which fig. 3A is an effect diagram of fixing the antenna at the inner side (exposing the outer side), fig. 3B is an effect diagram of fixing the antenna at the outer side (exposing the inner side), fig. 3C is an effect diagram of fixing the antenna at the back side (exposing the abdomen side), and fig. 3D is an effect diagram of fixing the antenna at the abdomen side (exposing the back side).
Detailed Description
The invention will be further described with reference to the drawings and specific examples. However, the present invention is not limited to the above-described embodiments, and various modifications may be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Examples
As shown in fig. 1 and 2, the directional fixing device of the irregular antenna of a living insect shape comprises a glass slide 1, an antenna pad table 6, an antenna pad table 3, a small water storage well device 2 and ultraviolet glue 5, wherein the small water storage well device 2 is fixedly arranged in the middle of the glass slide 1, the antenna pad table 6 is arranged on the glass slide 1, the antenna pad table 6 is positioned in the small water storage well device 2, the antenna pad table 3 is arranged on the glass slide 1, the antenna pad table 3 is positioned in the small water storage well device 2, the antenna pad table 3 is positioned at the side end of the antenna pad table 6 and is in contact with the side surface of the antenna pad table 6, the adhesive layer 7 is arranged on the upper surface of the antenna pad table 3, and the ultraviolet glue 5 is coated on the joint of the antenna pad table 6 and the antenna pad table 3;
the water storage small well device 2 is in a circular column or a hollow polygonal column;
the distance between one end of the adhesive layer 7 near the antenna pad stand and the antenna pad stand 6 is less than 1/2 of the length of the head of the insect, and the width of the adhesive layer 7 is consistent with the width of the insect pad stand 3;
the height of the water storage small well device 2 is 2-3 mm;
the width of the ultraviolet glue 5 is larger than the width of the insect head;
the insect body fixing device also comprises an insect body fixing line 4, wherein one end of the insect body fixing line 4 is fixedly arranged on the adhesive layer 7 of the insect body pad table 3; the insect fixing line 4 can better fix the insect bodies and avoid the struggling displacement of living insects;
the two insect body fixing lines 4 are respectively positioned at the head-chest section and the chest-abdomen section of the insect;
the length of the antenna pad table 6 is 5-20 mm, the width is 3-5 mm, and the height is 1-2 mm; the length of the insect pad table 3 is 10-25 mm, the width is 8-20 mm, and the height is 0.5-1 mm;
the water storage small well device is made of hot melt adhesive;
the antenna pad table 6 is a glass slide, and the worm pad table 3 is a cover glass;
the adhesive layer 7 is double-sided adhesive;
the antenna pad table can provide a base for directional fixation of the antenna, so that the height difference between the antenna and the insect body is convenient to coordinate, and the light transmittance is met; the insect body pad table is used for placing insect bodies and filling the height difference between the bodies and the antenna; the adhesive layer (double-sided adhesive) on the insect body pad table can reduce struggling of insect bodies, the insect body fixing line is arranged on the insect body pad table, the end of the insect body fixing line is fixedly adhered to the adhesive layer (double-sided adhesive), and the insect body fixing line is respectively pressed at the head-chest section and the chest-abdomen section to further fix the insect bodies; the connection part of the antenna pad table and the insect pad table and the ultraviolet adhesive on the antenna pad table can rapidly orient and fix the antenna and the head of the living insect without damaging the antenna structure and the function; the small well device is a water storage medium, can ensure that the type of the antenna sensor is determined under the magnification of 1000 times by means of a water objective lens (multiplied by 100), is positioned accurately, and an electrode is inserted into a specific sensor accurately by applying a microscopic operation technology;
the manufacturing method of the living insect irregular antenna directional fixing device comprises the following specific steps:
(1) Cutting the glass slide into an antenna pad table, and bonding the bottom surface of the antenna pad table to the middle part of another clean glass slide;
(2) Cutting the cover glass into an insect pad table, adhering the bottom surface of the insect pad table to the clean glass slide, and enabling the side end of the insect pad table to be in contact with the side surface of the antenna pad table;
(3) The adhesive layer is adhered and covered on the insect body pad table, and then the insect body fixing line is adhered to the adhesive layer of the insect body pad table, and the insect body fixing line is respectively positioned at the head-chest section and the chest-abdomen section of the insect;
(4) The ultraviolet glue is coated and arranged at the joint of the antenna pad table and the insect body pad table and on the antenna pad table, and the width of the ultraviolet glue is larger than the width of the insect body head;
(5) And a water storage small well device is arranged along the periphery of the antenna pad table and the insect pad table by adopting hot melt adhesive.
Examples
Directional fixation of living antenna for single sensor electrophysiological experiment of guava fruit fly of Bactrocera genus by adopting directional fixation device of irregular living antenna of example 1
The adult fruit fly of guava of Bactrocera is divided into three sections of head, chest and abdomen, and the total length is about 5-8mm; the tentacle of the guava fruit fly is in a mango shape and is divided into a handle section, a stem section and a whip section; the total length of the antenna is 900-1000 mu m, and the width of the whip is 100 mu m; the sensor is in an irregular geometric shape and very dense, and integrates sense organs with multiple functions of smell sense, hearing sense, temperature and humidity sense, gravity sense and the like; in order to fully study the working mode of the antenna, understand the electrophysiological response of different types of single sensors of the dorsal, ventral, medial and lateral antennas to external stimulus, the single sensors need to be oriented and fixed, and the specific steps are as follows:
(1) Cutting the glass slide into an antenna pad table with the length of 5mm multiplied by 3mm, and bonding the bottom surface of the antenna pad table to the middle part of the clean glass slide by using glue;
(2) Cutting the cover glass into insect body pad tables with the thickness of 10mm multiplied by 8mm, adhering the bottom surface of the insect body pad table to a clean glass slide, and enabling the side ends of the insect body pad table to be in contact with the side surfaces of the antenna pad table;
(3) The method comprises the steps of adhering an adhesive layer (double-sided adhesive) on an insect body pad table, adhering an insect body fixing line to the adhesive layer of the insect body pad table, and enabling the insect body fixing line to be respectively positioned at head-chest sections and chest-abdomen sections of insects; filling the height difference between the body of the guava fruit fly and the antenna by using the height difference between the cover glass and the glass slide;
(4) The method is characterized in that the method comprises the steps of placing the method under a split microscope, coating ultraviolet glue on the joint of an antenna pad table and an insect pad table and on the antenna pad table according to the requirement of an experimental target before fixing the antenna at a specific angle, enabling the width of the ultraviolet glue to be larger than the width of the insect head, enabling the width of the ultraviolet glue on the double-sided glue of the insect pad table to be about 2mm, and fixing the insect head and directional antenna conveniently;
(5) A water storage small well device is arranged along the periphery of the antenna pad table and the insect pad table by adopting hot melt adhesive; placing a small amount of water in the water storage small well device, and observing an antenna single sensor in the back side direction of the insect under the magnification of a high-power microscope and performing subsequent experimental operation;
when the inner side of the antenna of the fruit fly is fixed (the outer side is exposed), the feet and wings of the fruit fly of guava are sheared off under a split microscope; when the inner side of the antenna is to be fixed (the outer side is exposed), a small amount of sticky ultraviolet glue is smeared on the joint of the antenna pad table and the insect body pad table and on the antenna pad table, two sides of an insect body are gently clamped by using thin tweezers, and the abdomen sides of a living insect head, a chest and an abdomen are placed on the double-sided glue on the insect body pad table, so that the initial fixation of the insect body is completed; then, the abdomen part of the head is immersed into the ultraviolet glue on the double-sided glue of the insect pad table; under a split microscope, placing the inner side or the inclined inner side (the outer side is exposed) of the antenna into a small amount of ultraviolet glue by using a thin tweezer for preliminary fixation; after the completion, the ultraviolet glue is solidified by a special ultraviolet lamp, the ultraviolet lamp irradiates the position of the ultraviolet glue for 5 to 8 seconds, so that the ultraviolet glue is solidified, and the antenna is permanently fixed; in order to further fix the insect body, the insect body fixing line can be respectively arranged between the head and the chest and between the chest and the abdomen, and is tensioned and adhered with the double-sided adhesive tape to fix the insect body; adding a small amount of water into the water storage small well device, and observing an antenna single sensor in the back side direction of the insect under the amplification of a high-power microscope and performing subsequent experimental operation; the effect of fixing the inner side (exposing the outer side) of the antenna of the fly is shown in figure 3A;
when the outer side of the antenna of the fruit fly is fixed (the inner side is exposed), the feet and wings of the fruit fly of guava are sheared off under a split microscope; when the outer side of the antenna is to be fixed (the inner side is exposed), a small amount of sticky ultraviolet glue is smeared on the joint of the antenna pad table and the insect body pad table and on the antenna pad table, two sides of an insect body are gently clamped by using thin tweezers, and the abdomen sides of a living insect head, a chest and an abdomen are placed on the double-sided glue on the insect body pad table, so that the initial fixation of the insect body is completed; then, the abdominal part of the insect head is immersed in ultraviolet glue on the double-sided adhesive of the insect body pad table; under a split microscope, placing the outer side or the inclined outer side (the inner side is exposed) of the antenna into a small amount of ultraviolet glue by using a thin tweezer for preliminary fixation; after the completion, the ultraviolet glue is solidified by a special ultraviolet lamp, the ultraviolet lamp irradiates the position of the ultraviolet glue for 5 to 8 seconds, so that the ultraviolet glue is solidified, and the antenna is permanently fixed; in order to further fix the insect body, the insect body fixing line can be respectively arranged between the head and the chest and between the chest and the abdomen, and is tensioned and adhered with the double-sided adhesive tape to fix the insect body; adding a small amount of water into the water storage small well device, and observing an antenna single sensor in the inner side direction of the insect under the amplification of a high-power microscope and performing subsequent experimental operation; the effect of fixation (exposure) of the outer side of the tentacle of the fruit fly is shown in figure 3B;
when the back side of the antenna of the fruit fly is fixed (the abdomen side is exposed), the feet and wings of the fruit fly of guava are sheared off under a split microscope; when the back side of the antenna is to be fixed (the abdominal side is exposed), a small amount of sticky ultraviolet glue is smeared on the joint of the antenna pad table and the insect body pad table and on the antenna pad table, two sides of the insect body are gently clamped by using thin tweezers, and the abdominal sides of the living insect heads, the chest and the abdomen are placed on the double-sided glue on the insect body pad table, so that the initial fixation of the insect body is completed; then, the abdominal part of the insect head is immersed in ultraviolet glue on the double-sided adhesive of the insect body pad table; under a split microscope, the back side (the abdominal side is exposed) of the antenna is placed in a small amount of ultraviolet glue by using a thin tweezer for preliminary fixation; after the completion, the ultraviolet glue is solidified by a special ultraviolet lamp, the ultraviolet lamp irradiates the position of the ultraviolet glue for 5 to 8 seconds, so that the ultraviolet glue is solidified, and the antenna is permanently fixed; in order to further fix the insect body, the insect body fixing line can be respectively arranged between the head and the chest and between the chest and the abdomen, and is tensioned and adhered with the double-sided adhesive tape to fix the insect body; adding a small amount of water into the water storage small well device, and observing an antenna single sensor in the back side direction of the insect under the amplification of a high-power microscope and performing subsequent experimental operation; the effect of fixation of the back side of the tentacle of the fruit fly (ventral exposure) is shown in fig. 3C;
when the abdomen side of the bactrocera dorsalis is fixed (the back side is exposed), the feet and wings of the bactrocera guava are sheared under a split microscope; when the antennal abdominal side is to be fixed (back side is exposed), a small amount of sticky ultraviolet glue is smeared on the joint of the antennal pad table and the insect body pad table and on the antennal pad table, two sides of the insect body are gently clamped by using thin tweezers, and the abdominal sides of the living insect heads, the breasts and the abdomen are placed on the double-sided glue on the insect body pad table, so that the initial fixation of the insect body is completed; then the abdominal part of the insect head is immersed into the ultraviolet glue on the double-sided glue of the insect body pad table; under a split microscope, the abdomen side (back side exposure) of the antenna is placed into a small amount of ultraviolet glue by using a thin tweezer for preliminary fixation; after the completion, the ultraviolet glue is solidified by a special ultraviolet lamp, the ultraviolet lamp irradiates the position of the ultraviolet glue for 5 to 8 seconds, so that the ultraviolet glue is solidified, and the antenna is permanently fixed; in order to further fix the insect body, the insect body fixing line can be respectively arranged between the head and the chest and between the chest and the abdomen, and is tensioned and adhered with the double-sided adhesive tape to fix the insect body; adding a small amount of water into the water storage small well device, and observing an antenna single sensor in the back side direction of the insect under the amplification of a high-power microscope and performing subsequent experimental operation; the effect of the fixation of the fly antenna on the ventral side (dorsal exposure) is shown in fig. 3D.
The antenna pad table is used as a base for directional fixation of the antenna, so that the height difference between the antenna and the insect body is conveniently coordinated, and the light transmittance is met; the insect body pad table is used for placing insect bodies and filling the height difference between the bodies and the antenna; the adhesive layer (double-sided adhesive) on the insect body pad table can reduce struggling of insect bodies, the insect body fixing line is arranged on the insect body pad table, the end of the insect body fixing line is fixedly adhered to the adhesive layer (double-sided adhesive), and the insect body fixing line is respectively pressed at the head-chest section and the chest-abdomen section to further fix the insect bodies; the connection part of the antenna pad table and the insect pad table and the ultraviolet adhesive on the antenna pad table can rapidly orient and fix the antenna and the head of the living insect without damaging the antenna structure and the function; the small well device is a water storage medium, can ensure that the type of the antenna sensor is determined under the magnification of 1000 times by means of a water objective lens (multiplied by 100), is positioned accurately, and is inserted into a specific sensor accurately by applying a micromanipulation technology.
The directional fixing device for the irregular feeler of the living insect shape can realize the rapid and synchronous fixing of the feeler of the insect body and the extremely irregular morphology, especially can realize the directional fixing of the specific angle of the irregular feeler according to the position requirement of a single sensor experiment, meets the single sensor experiment microscopic operation of higher difficulty level, and solves the problems of the specific angle fixing, thicker feeler horny layer, low transmittance, easy target movement caused by the recording electrode puncture, and difficult accurate positioning operation due to the fact that the single sensor spacing is only 2-3 microns.
Claims (9)
1. The utility model provides a directional fixing device of irregular feeler of living body fruit fly class insect shape which characterized in that: the device comprises a glass slide (1), an antenna pad table (6), an insect body pad table (3), a water storage small well device (2) and ultraviolet glue (5), wherein the water storage small well device (2) is fixedly arranged in the middle of the glass slide (1), the antenna pad table (6) is arranged on the glass slide (1) and is positioned in the water storage small well device (2), the insect body pad table (3) is positioned at the side end of the antenna pad table (6) and is in contact with the side surface of the antenna pad table (6), an adhesive layer (7) is arranged on the upper surface of the insect body pad table (3), the ultraviolet glue (5) is arranged at the joint of the antenna pad table (6) and the antenna pad table (6), the antenna pad table (6) is formed by cutting the glass slide, and the bottom surface of the antenna pad table (6) is adhered to the other clean glass slide; the insect body pad table (3) is formed by cutting a cover glass, the bottom surface of the insect body pad table is adhered to the clean glass slide, and the side end of the insect body pad table is contacted with the side surface of the antenna pad table; the antenna pad table (6) is used as a base for directional fixation of the antenna, so that the height difference between the antenna and the insect body is conveniently coordinated, the insect body is placed by the insect body pad table (3), and the height difference between the insect body and the antenna is filled.
2. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the water storage small well device (2) is in a circular column or a hollow polygonal column.
3. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the distance between one end of the adhesive layer (7) close to the antenna pad table and the antenna pad table (6) is less than 1/2 of the length of the head of the insect, and the width of the adhesive layer (7) is consistent with the width of the insect pad table (3).
4. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the height of the water storage small well device (2) is 2-3 mm.
5. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the width of the ultraviolet glue (5) is larger than the width of the insect head.
6. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the pest pad also comprises two pest fixing lines (4), and one end of each pest fixing line (4) is fixedly arranged on the adhesive layer (7) of the pest pad table (3).
7. The device for fixing the orientation of irregular feelers of the shape of a living body fruit fly insect according to claim 1, characterized in that: the length of the antenna pad table (6) is 5-20 mm, the width is 3-5 mm, and the height is 1-2 mm; the length of the insect pad table (3) is 10-25 mm, the width is 8-20 mm, and the height is 0.5-1 mm.
8. The method for manufacturing the irregular antenna orientation fixing device for the living body fruit fly insects according to any one of claims 1 to 7, which is characterized by comprising the following specific steps:
(1) Cutting the glass slide into an antenna pad table, and bonding the bottom surface of the antenna pad table to the middle part of another clean glass slide;
(2) Cutting the cover glass into an insect pad table, adhering the bottom surface of the insect pad table to the clean glass slide, and enabling the side end of the insect pad table to be in contact with the side surface of the antenna pad table;
(3) The adhesive layer is adhered and covered on the insect pad table;
(4) The ultraviolet glue is coated and arranged at the joint of the antenna pad table and the insect body pad table and on the antenna pad table, and the width of the ultraviolet glue is larger than the width of the insect body head;
(5) And a water storage small well device is arranged along the periphery of the antenna pad table and the insect pad table by adopting hot melt adhesive.
9. The method for manufacturing the irregular antenna orientation fixing device for the living body fruit fly insects, which is characterized in that: and (3) adhering the insect body fixing line to the adhesive layer of the insect body pad table, and enabling the insect body fixing line to be respectively positioned at the head-chest section and the chest-abdomen section of the insect.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910625649.5A CN110286016B (en) | 2019-07-11 | 2019-07-11 | Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910625649.5A CN110286016B (en) | 2019-07-11 | 2019-07-11 | Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110286016A CN110286016A (en) | 2019-09-27 |
| CN110286016B true CN110286016B (en) | 2023-08-04 |
Family
ID=68021257
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910625649.5A Active CN110286016B (en) | 2019-07-11 | 2019-07-11 | Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110286016B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111458334B (en) * | 2020-04-16 | 2022-12-06 | 云南大学 | Visualization method for lymphatic vessels in light-color antennae of insects |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103115809A (en) * | 2013-01-23 | 2013-05-22 | 浙江大学 | Transmission electron microscope processing method for insect antenna samples |
| CN205357805U (en) * | 2015-12-16 | 2016-07-06 | 浙江大学 | Fixing device of small -size live body insect |
| CN206725851U (en) * | 2017-03-31 | 2017-12-08 | 中华人民共和国日照出入境检验检疫局 | One kind coordinates stereomicroscope multi-angle insect biology observation device |
| CN109141997A (en) * | 2018-08-03 | 2019-01-04 | 福建农林大学 | A kind of flaking method of microexamination Aphytis |
| CN208725934U (en) * | 2017-09-08 | 2019-04-12 | 四川大学华西医院 | A kind of fixed device of living animal micro-imaging |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1717628B1 (en) * | 2004-02-16 | 2012-01-25 | Olympus Corporation | Immersion objective lens, retention mechanism for immersion medium and manufacturing method thereof |
| US8865630B2 (en) * | 2010-02-26 | 2014-10-21 | Massachusetts Institute Of Technology | High-throughput platform for in-vivo sub-cellular screens on vertebrate larvae |
| US10802262B2 (en) * | 2015-10-29 | 2020-10-13 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and systems for imaging a biological sample |
-
2019
- 2019-07-11 CN CN201910625649.5A patent/CN110286016B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103115809A (en) * | 2013-01-23 | 2013-05-22 | 浙江大学 | Transmission electron microscope processing method for insect antenna samples |
| CN205357805U (en) * | 2015-12-16 | 2016-07-06 | 浙江大学 | Fixing device of small -size live body insect |
| CN206725851U (en) * | 2017-03-31 | 2017-12-08 | 中华人民共和国日照出入境检验检疫局 | One kind coordinates stereomicroscope multi-angle insect biology observation device |
| CN208725934U (en) * | 2017-09-08 | 2019-04-12 | 四川大学华西医院 | A kind of fixed device of living animal micro-imaging |
| CN109141997A (en) * | 2018-08-03 | 2019-01-04 | 福建农林大学 | A kind of flaking method of microexamination Aphytis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110286016A (en) | 2019-09-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lees | The sensory physiology of the sheep tick, Ixodes ricinus L | |
| Knudsen | Distinct auditory and lateral line nuclei in the midbrain of catfishes | |
| Sterbing-D'Angelo et al. | Bat wing sensors support flight control | |
| Xu et al. | Design and fabrication of a high-density metal microelectrode array for neural recording | |
| Krüger et al. | Simultaneous recording with 30 microelectrodes in monkey visual cortex | |
| Jiang et al. | Hydroprinted liquid‐alloy‐based morphing electronics for fast‐growing/tender plants: from physiology monitoring to habit manipulation | |
| KR101007558B1 (en) | A flexible, multi-channel microelectrode for recording laboratory animal EEG and method for recording laboratory animal EEG using the same | |
| AU2014329510A1 (en) | Organ mounted electronics | |
| CN110786846B (en) | Flexible recording electrode and preparation method and implantation method thereof | |
| RU2524134C2 (en) | Device, containing cardiomyocytes, and method of production and application thereof | |
| JP2022508575A (en) | Measurement system | |
| Rountree et al. | Prototype chemical synapse chip for spatially patterned neurotransmitter stimulation of the retina ex vivo | |
| WO2017173339A1 (en) | Implantable medical devices for optogenetics | |
| KR100466954B1 (en) | Apparatus and method for screening, olfactory mucosa stimulating compound found by the screening method, and therapeutic apparatus and electrode section for measurement | |
| CN110286016B (en) | Irregular antenna directional fixing device for living fruit fly insects and manufacturing method thereof | |
| Goris et al. | Central response to infra-red stimulation of the pit receptors in a crotaline snake, Trimeresurus flavoviridis | |
| JP2022508576A (en) | Electrode equipment | |
| Fiáth et al. | Fine-scale mapping of cortical laminar activity during sleep slow oscillations using high-density linear silicon probes | |
| Jiao et al. | Scalable fabrication framework of implantable ultrathin and flexible probes with biodegradable sacrificial layers | |
| CN113941378A (en) | Multi-cavity electrophysiological micro-nano detection-based neural organoid chip and detection method | |
| CN210487421U (en) | Directional fixing device of irregular antenna of living insect shape | |
| Manley et al. | Reversed tonotopic map of the basilar papilla in Gekko gecko | |
| Canfield et al. | Methods for chronic neural recording in the telencephalon of freely behaving fish | |
| DE10142393C1 (en) | Apparatus measuring bioelectrical signals from spheroids, includes commensurately-sized chamber ringed internally with coplanar electrodes | |
| Márton et al. | In vivo measurements with robust silicon-based multielectrode arrays with extreme shaft lengths |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |