KR101056581B1 - Automated detection system for stable behavior reaction of linear pretty nematodes and similar species under stable conditions by blocking leakage of volatile toxic compounds - Google Patents

Abstract

The present invention relates to a stable condition reaction behavior, and more particularly, to devise an appropriate blocking device during the experiment to maintain a constant concentration of VOC (Volatile Organic Compound) and to be tested under standardized conditions and to be toxic to the human body. The present invention relates to an automated detection system for the stable condition reaction behavior of linear nematodes and similar species under stable conditions through the blocking of volatile toxic compounds which protects the health and laboratory environment of the investigator from a volatile VOC material.

Such a feature of the present invention is a transparent box that is provided with a microscope and the camera mounted on the fortified slide is sealed inside; A computer connected to the microscope in the transparent box; A monitor connected to the computer; And it is characterized in that it comprises a supply sealed box for supplying the VOC into the transparent box.

Volatile, Toxic Compounds, Pretty Nematodes, Reaction

Description

AUTOMATIC DETECTION SYSTEM OF RESPONSE BEHAVIORS OF THE LINE-SHAPED Caenoraptis elegans AND RELATED SPECIES UNDER STABLE CONDITIONS BY BLOCKING LEAKAGE OF VOLATILE TOXIC SUBSTANCES}

The present invention relates to an automatic reaction behavior detection system, and more particularly, to design a suitable blocking device to maintain a constant concentration of VOC during the experiment and to be tested under standardized conditions and to be toxic to human body The present invention relates to an automated detection system for stable behavior reaction of linear nematodes and similar species under stable conditions through the blocking of volatile toxic compounds to protect the health and laboratory environment of the investigator.

In general, VOC materials such as benzene, toluene and formaldehyde are organic compounds and are well volatile, making it difficult to maintain VOC concentrations for a long time. However, if constant concentrations are not maintained, it is difficult to perform standardized experiments.

In addition, volatile toxic substances are toxic to humans, so they can be dangerous to the tester or the occupant if exposed and handled externally.

Due to the difficulty and stability of the above test conditions, only physicochemical and toxicological analyzes were performed under strictly standardized test conditions, and behavioral risk assessment was difficult.

The present invention for solving the above problems is to devise an appropriate blocking device during the experiment to maintain a constant concentration of VOC and to be tested under standardized conditions.

And since the behavioral response must be acquired as a video image, it produces a noise-free image.

It also protects the health of the investigator and the laboratory environment from volatile VOCs that are toxic to humans.

Furthermore, the experimental animal is linear, so apply the analysis program applied to the linear movement.

In order to achieve the above object, a stable condition reaction behavior automatic detection system of a linear pretty kid nematode and similar species under stable conditions through the blocking of volatile toxic compounds according to the present invention, the camera 11 and the fortified slide inside A sealed transparent box 13 in which a microscope 12 to which the 20 is mounted is sealed; A computer (14) connected to the microscope (12) in the hermetic transparent box (13); A monitor 15 connected with the computer 14; And a supply sealed box 16 for supplying a volatile organic compound (VOC) into the sealed transparent box 13.

Thus, the citadel-type slide 20, the hermetic transparent case 21 is located outside; A base 22 located inside the transparent case 21; An observation area (23) positioned at the center of the base (22) to place micro-animals; An observation stage 24 on which the observation region 23 is located; And a furrow region 25 formed around the observation stage 24 and having a groove formed therein.

In addition, the micro-animals may be provided with a linear pretty nematode and similar species, the volatile organic compound (VOC, Volatile Organic Compound) may include a compound such as benzene, toluene, formaldehyde and any one or more compounds It can be configured as.

In addition, a water barrier 27 may be formed between the upper portion of the viewing area 23 positioned in the slide 20 and the cover glass 26 of the slide 20.

The present invention configured as described above maintains a constant VOC concentration in the observation region during the experiment.

It also helps maintain the health of the experimenter by preventing unnecessary exposure of VOCs to the laboratory environment.

In addition, it generates images without image noise and detects response behavior by analyzing linear images.

Hereinafter, with reference to the accompanying drawings will be described in detail.

1 is a schematic configuration diagram of a stable condition reaction behavior automatic detection system according to the present invention, Figure 2 is a perspective view of a fort-type slide of the stable condition reaction behavior automatic detection system according to the present invention, Figure 3 is 4 is a plan view of the fort-type slide of the stable condition reaction behavior automatic detection system, and FIG.

In addition, Figure 5 is a test example of the linear motion of the nematode before the treatment of toxic substances in the automatic stable reaction reaction behavior detection system according to the present invention, Figure 6 is a toxic substance (benzene 1ppm in the automatic detection reaction conditions stable behavior according to the present invention) Exemplary line traces of the reaction behavior of nematodes ( C. elegans ) after treatment, FIG. 7 shows the pretreatment (light color) and after treatment of benzene (0.1 ppm) in the automatic detection system for stable reaction reaction behavior according to the present invention. Fig. 8 is a diagram illustrating the comparison of the main parameters of (dark color), and FIG. 8 shows the main parameters before (light) and after (dark) the treatment of toluene (0.1 ppm) in the stable condition reaction behavior automatic detection system according to the present invention. 9 is an illustration of the comparison of the main parameters before (light) and after (dark color) of formaldehyde (0.1 ppm) in the stable condition reaction behavior automatic detection system according to the present invention And Figure 10 is in the present invention Other stable reaction conditions, in the automatic detection system behavior benzene, toluene, Pro aldehyde is an exemplary view showing that each of the comparison of the main parameters (each 0.1ppm) mixture before treatment (light color) and after treatment (dark).

That is, the stable detection reaction behavior of the linear pretty little nematode and the similar species under stable conditions through the blocking of volatile toxic compounds according to the present invention, as shown in Figures 1 to 10, the camera ( 11) and a closed transparent box 13 provided with a microscope 12 on which the fortified slide 20 is mounted.

In addition, a computer 14 connected to the microscope 12 in the sealed transparent box 13 and a monitor 15 connected to the computer 14 are provided.

Particularly, in the stable condition reaction behavior automatic detection system 10 according to the present invention, a supply sealing box 16 for supplying a volatile organic compound (VOC) to the sealed transparent box 13 is further included. It is provided.

Furthermore, the citadel-type slide 20 is composed of a transparent case 21 positioned to the outside, so that the inside of the transparent case 21 can be observed by the naked eye or by a microscope or a camera.

In addition, the base 22 positioned inside the transparent case 21 and the observation area 23 positioned at the center of the base 22 to place linear micro-animals (for example, linear pretty nematodes and similar species) ) Is prepared.

In addition, an observation stage 24 in which the observation region 23 is located is provided, and a furrow region 25 having a groove formed around the observation stage 24 is provided. The solution is filled.

In particular, a water barrier 27 is formed between the upper portion of the viewing area 23 positioned in the slide 20 and the cover glass 26 of the slide 20.

Looking at the embodiment for the stable condition reaction behavior automatic detection system 10 according to the present invention is provided as follows.

(1) Small linear micro-animals such as nematodes ( C. elegans ) were able to detect the reaction behavior under a microscope at a stable concentration without volatile substances such as volatile organic compounds harmful to the environment (Fig. 1) . Observation chamber (80 × 45 × 40 cm) for storing microscopes with fortified slides and cameras (“13” in Fig. 1) and VOC boxes for supplying volatile toxic substances from the outside into sealed transparent boxes (15 × 15 × 10 cm) (“16” in FIG. 1) and a computer and monitor (“14” in FIG. 1) connected to the camera.

(2) The gist of the present invention is that the fortress-type slide (Fig. 2) is manufactured and the observation area is double sealed with a water film, so that the volatile toxic substances in the observation area are stably observed without volatilization. . The citadel-type slide (Fig. 2) has an observation stage (Fig. 2) that can contain a test animal and a solution and a small furrow surrounding it. On the observation stage there is a round observation area with a diameter of 1 mm (No. 23 in Figs. 2, 3 and 4) where the test solution and test organisms are placed. Surrounding the observation stage is a small trench 3 mm deep and 3 mm wide (No. 25 in Fig. 4), filled with test solution. For convenience, you can create a number of fortified stages. In this case there are three fortified stages (FIG. 2).

(3) After filling the observation area and the furrow surrounding the observation stage with the test organism and the solution, cover glass (No. 26 in Fig. 4) is covered with no bubbles. A water film (No. 27 in Fig. 4) is formed due to the surface tension of the water in the boundary section surrounding the curved glass and the viewing area. This water film prevents the rapid volatilization of volatiles. Since there is a sealed container on the outside and the sealed container contains a certain concentration of toxic substances, it is balanced between the partial pressure of the toxic substance in the long-term sealed container and the toxic substance partial pressure in the observation area to maintain a stable concentration. External furrows can be doubled or tripled for more complete volatile protection. Citadel slides are installed on a petri dish containing water to maintain temperature (FIG. 2).

(4) VOCs (Volatile Organic Compounds) are sealed boxes used to supply volatile toxicants (16). If the liquid volatile substance is contained in the VOC box, the VOC box is sealed and the empty space in the VOC box is saturated with toxic substances. After the preparation for observation under the microscope is completed, the saturated toxic substance is injected into the sealed transparent box 13 with the microscope through the silicon tube by adjusting the valve.

(5) The sealed transparent box is sealed so that the toxic substances supplied from the VOC box do not go out of the observation system, and the concentration of the toxic substances in the whole observation system is kept stable. It also ensures that no toxic substances are exposed to the outside environment. At the end of the experiment, the outlet on the sealed box allows for safe release of toxic substances.

(6) On the microscope, the camera which can observe the shape of the test animal and the image of the image can be traced and the program which can recognize the abnormal reaction is installed.

A) In the present invention, maintaining a stable concentration of the entire observation system using a sealed transparent box and block unnecessary exposure.

1) First of all, in one embodiment of the hermetic transparent box 13, this box is made of acrylic material (1 cm thick, 45 cm wide, 40 cm high, 80 cm high) which is used to seal the microscope and observation stage connected to the camera from the outside ( See FIG. 1). Make a sealed hole around the box to connect the microscope connection (microscope and camera power cord, video signal cord; 2.5cm in diameter) and the VOC box (2cm in diameter).

2) Install a tube (10 cm in diameter) to release the gas on the roof of the observation box (Figure 1).

B) Stable toxic substance supply through VOC box (16)

1) The supply sealed box 16, which is a VOC box, was made of acrylic (1cm thick, 15cm wide x 15cm high, 10cm high). Make a sealed hole with a diameter of 2 cm and connect it to a sealed transparent box with a rubber tube.

2) The supply closure box (16), a VOC box, contains a liquid toxic substance solution. At room temperature (20-30 ° C) VOC liquid automatically evaporates to produce gas. This gas is saturated in the VOC box.

3) Saturated gas is passed through a rubber tube to a sealed transparent box.

C) Maintaining stable concentration and removing image noise by implementing star-shaped observation stage and observation area

1) Acrylic, 6cm * 3.5cm wide, 0.5cm high. There is an observation area with a diameter of 4mm and a depth of 1.5mm on the observation stage. Around the viewing area is a furrow 2 mm deep (FIG. 2). In this case, three observation stages were made. One of the observation areas was selected and covered with a cover glass.

2) If the solution is filled in the furrow and the observation area and the cover glass is covered, a thin water film is formed between the observation stage and the cover glass due to the surface tension of the water. This blocks the rapid volatilization of volatiles. Equilibrium of the toxicant partial pressure in the sealed transparent box and the toxicant partial pressure in the viewing area is maintained to maintain a stable concentration of the toxic material in the stable viewing area.

3) Covering the cover glass prevents the diffusion of volatile materials as well as blocking the reflection of images from the solution to remove image noise.

4) Temperature maintenance through cooling petri dish

i. Fill the water to the appropriate height using 10 cm (diameter) x 1.5 cm (height) Petri dishes (FIG. 2).

ii. Petri dishes filled with water lower the rising temperature due to microscopic light. The amount of water can be appropriately adjusted according to the light intensity and the observation period.

la. Observation process

1) The movement of the test animal without noise is traced through a linear imaging device.

2) Reaction behavior assays after treatment.

Figure 5 relates to the linear movement of nematodes ( C. elegans ) before toxic treatment. Figure 6 relates to the line trajectory of the reaction behavior of nematodes ( C. elegans ) after treatment with toxic substances (1 ppm benzene).

Each segment of the linear line is divided into 13 segments and the speed, acceleration, stop, turnover rate and radius of curvature for each segment are calculated before and after processing. Each parameter is different from before and after appropriately. As can be seen from FIG. 7, when 1 ppm of benzene was treated, the stop time, the number of stops, the rotation rate, the radius of curvature, and the like after the treatment (dark color) were significantly reduced compared to the pretreatment (light color). 8 shows the reaction when 1 ppm of toluene is treated. The parameters decreased, similar to the case of treatment with benzene. However, compared to benzene, it can be seen that the speed and acceleration are also greatly reduced. By properly comparing the variations of the parameters, differences in compound quality can also be determined. Other parameters such as formaldehyde (FIG. 9), and drugs mixed with benzene, toluene and formaldehyde (FIG. 10) also showed differences in parameters.

The embodiments of the present invention have been described in detail above, but since the embodiments have been described so that those skilled in the art to which the present invention pertains can easily carry out the present invention, The technical spirit of the present invention should not be interpreted limitedly.

1 is a schematic diagram of a stable condition response behavior detection system according to the present invention.

Figure 2 is a perspective view of the fort slide of the stable condition reaction behavior detection system according to the present invention.

Figure 3 is a plan view of the fort type slide of the stable condition response behavior detection system according to the present invention.

Figure 4 is a side view of the fort slide of the stable condition response behavior detection system according to the present invention.

Figure 5 is a test example of the linear movement of the nematode before the treatment of toxic substances in the stable condition response behavior detection system according to the present invention.

Figure 6 is an illustration of the line trajectory of the reaction behavior of nematodes ( C. elegans ) after the treatment of toxic substances (benzene 1ppm) in the stable condition reaction behavior automatic detection system according to the present invention.

Figure 7 is an illustration of the comparison of the main parameters before (light) and after the treatment (dark color) of benzene (0.1 ppm) in the stable condition reaction behavior automatic detection system according to the present invention.

8 is an exemplary diagram for comparing the main parameters before (light) and after (dark) the treatment of toluene (0.1ppm) in the stable condition reaction behavior automatic detection system according to the present invention.

Figure 9 is an illustration of the comparison of the main parameters before (light) and after the treatment (produced) of the prohmaldehyde (0.1ppm) in the stable condition reaction behavior automatic detection system according to the present invention.

10 is an exemplary diagram for comparing the main parameters before (light) and after (dark) the treatment of benzene, toluene, proaldehyde (each 0.1ppm) mixture in the stable condition reaction behavior automatic detection system according to the present invention.

<Description of the symbols for the main parts of the drawings>

10: Stable Condition Response Behavior Detection System

11: Observation apparatus with a camera 12: Microscope

13: sealed transparent box 14: computer

15 monitor 16 supply sealed box

20: citadel slide 21: transparent case

22: base 23: observation area

24: Observation stage 25: furrow area

26: cover glass 27: water film

Claims (5)

A sealed transparent box 13 in which a microscope 12 equipped with a camera 11 and a fortified slide 20 is sealed therein; A computer (14) connected to the microscope (12) in the hermetic transparent box (13); A monitor 15 connected with the computer 14; And Supply sealed box 16 for supplying a volatile organic compound (VOC, Volatile Organic Compound) into the transparent transparent box 13; linear in a stable condition through the blocking of volatile toxic compounds, characterized in that it is provided Automated Detection System for Stable Condition Responses of Pretty Nematodes and Similar Species in Korea. The method of claim 1, The fortified slide 20, A transparent case 21 positioned outside; A base 22 located inside the transparent case 21; An observation area (23) positioned at the center of the base (22) to place micro-animals; An observation stage 24 on which the observation region 23 is located; And A furrow region 25 having a groove formed around the observation stage 24; Stable condition behavior of automatic detection of the reaction conditions of the linear pretty little nematodes and similar species under stable conditions through the blocking of volatile toxic compounds, characterized in that it is provided. 3. The method of claim 2, The micro-animal system for detecting stable reactions of linear pretty little nematodes and similar species under stable conditions through the blocking of volatile toxic compounds, characterized by being provided with linear pretty little nematodes and similar species. 3. The method of claim 2, A water barrier 27 is formed between the upper portion of the observation area 23 located in the slide 20 and the cover glass 26 of the slide 20. Automatic detection system for stable behavior response of linear pretty nematodes and similar species under stable conditions through blocking. 5. The method according to any one of claims 1 to 4, The volatile organic compound (VOC) is a linear, pretty nematode and similar species under stable conditions through the blocking of volatile toxic compounds, characterized in that it comprises any one or more compounds of benzene, toluene, formaldehyde. Automatic detection system for stable behavior of reaction conditions.

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